CN1823168A - 提高植物非生物胁迫耐受性和/或生物量的方法及用该方法所产生的植物 - Google Patents
提高植物非生物胁迫耐受性和/或生物量的方法及用该方法所产生的植物 Download PDFInfo
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Abstract
提供了提高植物对非生物胁迫的耐受性和/或增加植物的生物量的多核苷酸序列及其使用方法。
Description
发明领域和背景
本发明涉及提高植物非生物胁迫耐受性和/或植物生物量的方法,更具体地说,本发明涉及表达外源非生物胁迫-耐受性基因的植物。
非生物胁迫(也称为“环境胁迫”)条件例如高盐、干旱、洪涝、亚适温度和有毒化学物质污染,严重损害农作物。大多数植物有保护它们自己不受这些条件影响的进化策略。然而,如果这些胁迫条件太严重并且持续太久,则对植物的发育、生长和大多数作物的产量影响巨大。此外,大多数作物对非生物胁迫(ABS)十分敏感,因而为了经济作物的产量就需要提供最适合的生长条件。连续处于胁迫而引起植物代谢的重大改变,将最终导致细胞死亡和产量下降。因此,纵有广泛深入研究和采用复杂的和强化的作物保护措施,非生物胁迫条件造成的损失每年仍然以10亿美元计(1,2)。
开发胁迫耐受性植物至少是解决或调解某些这类问题的可能的策略。然而,用于开发新的耐受ABS的植物品系的传统植物育种策略,因其缓慢、耗时和结果难料,所以效率相对低下。此外,有限的胁迫耐受性的种质资源和远缘植物种间杂交的不相容性是传统育种中遇到的重大难题。另外,导致ABS耐受性的细胞过程本质上是复杂的而且包括细胞适应的多种机制和许多代谢途径(4-7)。
旨在使转基因作物具有非生物胁迫耐受性的遗传工程研究,在现有技术中已有描述。Apse和Blumwald(Curr Opin Biotechnol.13:146-150,2002)、Quesada等(Plant Physiol.130:951-963,2002)、Holmstrm等(Nature 379:683-684,1996)、Xu等(Plant Physiol110:249-257,1996)、Pilon-Smits和Ebskamp(Plant Physiol 107:125-130,1995)和Tarczynski等(Science 259:508-510,1993)的许多研究全都试图获得胁迫耐受性的植物。
此外,一些美国专利和专利申请也描述了与胁迫耐受性相关的多核苷酸及其在获得胁迫耐受性植物中的应用。美国专利号5,296,462和5,356,816描述了用编码参与拟南芥(Arabidopsis thaliana)冷适应的蛋白的多核苷酸转化植物,因而能促进转化植物的冷耐受性。
美国专利号6,670,528描述了用编码与胁迫效应元件结合的多肽的多核苷酸转化植物,因而能促进转化植物对非生物胁迫的耐受性。
美国专利号6,720,477描述了用编码信号转导胁迫相关蛋白的多核苷酸转化植物,因而能够提高转化植物对非生物胁迫的耐受性。
美国申请顺序号09/938842和10/342224描述了非生物胁迫相关基因及其在赋予植物对非生物胁迫的耐受性中的应用。
美国申请顺序号10/231035描述了植物过量表达钼辅因子硫化酶,因而提高其对非生物胁迫的耐受性。
虽然上述研究至少部分成功地获得了胁迫耐受性植物,但仍需胁迫耐受性基因以获得广泛耐受非生物胁迫条件的植物。
在将本发明付诸实施时,发明人通过生物信息学和实验室研究业已证实了一些新的非生物胁迫耐受性基因,这些新基因能提高植物的非生物胁迫耐受性和/或生物量。
发明概述
按照本发明的一方面,提供提高植物对非生物胁迫的耐受性的方法。该方法包括在植物中表达外源多核苷酸,该外源多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有90%同源。
按照本发明的另一方面,提供提高植物对非生物胁迫的耐受性的方法。该方法包括在植物中表达其外源多肽,该外源多肽包括选自SEQ ID NO:39-92的氨基酸序列。
按照本发明的又一方面,提供提高植物生物量的方法。该方法包括在植物中表达外源多核苷酸,该外源多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有90%同源。
按照本发明的再一方面,提供提高植物生物量的方法。该方法包括在植物中表达外源多肽,该外源多肽包括选自SEQ ID NO:39-92的氨基酸序列。
按照本发明的再又一方面,提供包含外源多核苷酸的植物细胞,该多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有90%同源。
按照本发明的再另一方面,提供包含外源多核苷酸的植物细胞,该外源多核苷酸编码包括选自SEQ ID NO:39-92的氨基酸序列的多肽。
按照本发明的再另一方面,提供核酸构建体,该核酸构建体包含与选自SEQ ID NO:1-18的核苷酸序列至少有90%同源的多核苷酸和能够在宿主细胞内指导所述多核苷酸转录的启动子。
按照本发明的再另一方面,提供核酸构建体,该核酸构建体包括编码包含选自SEQ ID NO:39-92的氨基酸序列的多肽的多核苷酸和能够在宿主细胞内指导所述多核苷酸转录的启动子。
按照本发明的还又一方面,提供分离的多肽,该多肽所含的氨基酸序列与选自SEQ ID NO:1-18的多核苷酸编码的氨基酸序列至少有90%同源。
按照本发明的另一方面,提供分离的多肽,该多肽包括选自SEQID NO:39-92的氨基酸序列。
按照下述的本发明优选实施方案的进一步特征,通过下述步骤来实现表达:(i)用外源多核苷酸转化植物的细胞;(ii)从所述细胞产生成熟植株;(iii)在适合所述成熟植株中表达所述外源多核苷酸的条件下,栽培所述成熟植株。
按照所述优选实施方案的再进一步特征,转化是通过将核酸构建体导入植物细胞中来实现的,该构建体包括外源多核苷酸和至少一个能够在植物细胞内指导外源多核苷酸转录的启动子。
按照所述优选实施方案的再进一步特征,所述至少一个启动子是组成型启动子。
按照所述优选实施方案的再进一步特征,所述组成型启动子是CaMV 35S启动子。
按照所述优选实施方案的再进一步特征,所述组成型启动子是At6669启动子。
按照所述优选实施方案的再进一步特征,所述至少一个启动子是诱导型启动子。
按照所述优选实施方案的再进一步特征,所述诱导型启动子是非生物胁迫诱导型启动子。
按照所述优选实施方案的再进一步特征,所述至少一个启动子是组织特异性启动子。
按照所述优选实施方案的再进一步特征,所述表达是通过用包括外源多核苷酸的病毒感染植物来实现的。
按照所述优选实施方案的再进一步特征,所述病毒是无毒病毒。
按照所述优选实施方案的再进一步特征,所述非生物胁迫选自高盐、失水、低温、高温、重金属毒性、无氧生活、营养缺乏、营养过剩、大气污染和UV辐射。
按照所述优选实施方案的再进一步特征,所述植物是双子叶植物。
按照所述优选实施方案的再进一步特征,所述植物是单子叶植物。
按照所述优选实施方案的再进一步特征,所述植物细胞成为植物的一部分。
本发明通过提供利用新的非生物胁迫耐受性基因提高植物非生物胁迫耐受性和/或生物量的方法,成功地克服了现有技术的缺点。
附图简述
本文仅通过实施例并参考附图描述本发明,下面具体地参考附图的细节,须强调的是,所给出的具体细节仅仅作为实例,用于说明性地论述本发明的优选实施方案,并且提供这些详细资料是为了提供所认为的对本发明的原理和构思最有用最容易理解的描述。在这方面,并没有试图以比基本理解本发明所必需的更为详细的说明本发明的结构细节,从附图获取的说明,使得在实践中如何使本发明的所述几种形式具体化对于本领域技术人员而言显而易见的。
图1是说明从核酸序列数据库中鉴定推定植物胁迫耐受性基因的流程图。
图2A-B是说明开花期的T2转基因拟南芥成熟植株用At6669启动子表达外源萤光素酶转基因的照片。相同植物在正常光照条件下(图2A)和在暗处(图2B)。在花组织和根组织中观察到证明萤光素酶表达的强发光。
图3说明在正常条件或胁迫条件下(分别用0或100M NaCl溶液灌溉)生长的转基因T1拟南芥(A.thaliana)植物的平均鲜重。用推定胁迫耐受性基因,或者用萤光素酶报道基因(对照)转化植物,所述基因处于At6669启动子的转录控制下。按照单向ANOVA T检验,平均值±SD没有显著性差异。
图4A说明在正常条件或胁迫条件下(分别用0或100M NaCl溶液灌溉)生长的转基因T2拟南芥植物的平均鲜重。用本发明推定胁迫耐受性基因,或者用萤光素酶报道基因(对照)转化植物,所述基因处于35S启动子的转录控制下。按照单向ANOVA T检验,平均值±SD没有显著性差异。
图4B说明在正常条件或胁迫条件下(分别用0或100M NaCl溶液灌溉)生长的转基因T2拟南芥植物的平均鲜重。用本发明的推定胁迫耐受性基因,或者用萤光素酶报道基因(对照)转化植物,所述基因处于At6669启动子的转录控制下。按照单向ANOVA T检验,平均值±SD没有显著性差异。
图5说明在盐胁迫条件下(用100mM NaCl溶液灌溉)生长的转基因T1拟南芥植物与在正常条件下(只用水灌溉)生长的类似植物比较的相对鲜重(%)。用本发明推定胁迫耐受性基因,或者用萤光素酶报道基因(对照)转化植物,所述基因处于At6669启动子的转录控制下。
优选实施方案的描述
本发明是涉及利用新的非生物胁迫耐受性基因提高植物非生物胁迫耐受性和/或生物量的方法,并且涉及显示出提高了对胁迫条件的耐受性和/或提高了累积生物量的植物。
参考附图及附加说明可以更好地理解本发明的原理和操作。
在详细解释本发明的至少一个实施方案之前,应该理解,本发明不受下述描述或附图说明中成分的构建和安排细节的限制。本发明可以有其他实施方案,或者说可用各种方法实践或完成本发明。同样,应该理解,本文所用的用语和术语是用来描述而非限制。
当将本发明付诸实施时,发明人应用生物信息学技术鉴定了编码推定非生物-胁迫耐受性(ABST)蛋白的多核苷酸序列(实施例1)。分离所选择的序列(实施例2),克隆到表达载体中(实施例3-4),再导入拟南芥植物中(实施例5)。这些在盐胁迫条件下或者在正常条件下生长的植物,与没有携带外源ABST基因的类似植物比较,都表现出显著更高的生物量(实施例6)。
因此,按照本发明的一个方面,提供提高植物对非生物胁迫的耐受性和/或植物生物量的方法。该方法包括在植物中表达外源多核苷酸,该外源多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有70%同源,优选至少有80%同源,更优选至少有85%同源,最优选至少有90%同源。或者,本发明的外源多核苷酸编码具有选自SEQ ID NO:39-92的氨基酸序列的多肽。
本文所用的术语“非生物胁迫”是指对植物的代谢、生长、繁殖和/或生存力的任何负面影响。因此,非生物胁迫能够被诸如以下的亚适环境生长条件所诱导:高盐、失水、洪涝、低温或高温、重金属毒性、无氧生活、营养缺乏、大气污染或UV辐射。
本文所用的术语“非生物胁迫耐受性”是指植物承受非生物胁迫时,其代谢、生长、繁殖力和/或生存力无重大改变的能力。
适用本发明方法的植物可以是任何单子叶植物或双子叶植物,包括但不限于玉米、小麦、大麦(barely)、黑麦、燕麦、水稻、大豆、花生、豌豆、小扁豆和苜蓿、棉花、油菜籽、双低油菜(canola)、胡椒、向日葵、马铃薯、烟草、番茄、茄子、桉树、树木、观赏植物、多年生牧草、饲料作物。
本文所用的术语“外源多核苷酸”是指在植物中不会自然表达的核酸序列,但是当将其导入植物中会以稳定或瞬时的方式产生至少一种多肽产物。
在植物中表达本发明的外源多核苷酸,可通过下述方法付诸实施:首先用外源多核苷酸转化植物的一种或多种细胞,其次从转化细胞产生成熟植株,最后在适合所述成熟植株中表达所述外源多核苷酸的条件下,栽培所述成熟植株。
优选转化是通过将核酸构建体导入植物细胞中来实现,该构建体包含本发明的外源多核苷酸和至少一个能在植物细胞内指导外源多核苷酸转录的启动子。下文将提供适合的转化方法的进一步详细说明。
本文所用的术语“启动子”是指基因转录起始位点上游的DNA区,RNA聚合酶与之结合后启动RNA的转录。启动子控制基因在何处(例如植物的哪一部分、动物的哪一器官等等)和/或何时(例如生物体一生的哪一阶段或状态)表达。
本发明的核酸构建体可采用任何合适的启动子序列。优选启动子是组成型启动子、组织特异性启动子或非生物胁迫诱导型启动子。
合适的组成型启动子包括例如CaMV 35S启动子(SEQ ID NO:19;Odell等,Nature 313:810-812,1985);拟南芥At6669启动子(SEQ IDNO:20);玉米Ubi 1(Christensen等,plant Sol.Biol.18:675-689,1992);水稻肌动蛋白(McElroy等,Plant Cell 2:163-171,1990);pEMU(Last等,Theor.Appl.Genet.81:581-588,1991);合成的Super MAS(Ni等,The Plant Journal 7:661-76,1995)。其它组成型启动子包括在以下美国专利号中的那些启动子:5,659,026、5,608,149、5,608,144、5,604,121、5,569,597、5,466,785、5,399,680、5,268,463和5,608,142。
合适的组织特异性启动子包括但不限于由例如以下文献所描述的叶特异性启动子:Yamamoto等,Plant J.12:255-265,1997;Kwon等,Plant Physiol.105:357-67,1994;Yamamoto等,Plant Cell Physiol.35:773-778,1994;Gotor等,Plant J.3:509-18,1993;Orozco等,Plant Mol.Biol.23:1129-1138,1993;Matsuoka等,Proc.Natl.Acad.Sci.USA 90:9586-9590,1993。
合适的非生物胁迫诱导型启动子包括但不限于盐诱导型启动子,例如RD29A(Yamaguchi-Shinozalei等,Mol.Gen.Genet.236:331-340,1993);干旱诱导型启动子,例如玉米rab17基因启动子(Pla等,Plant Mol.Biol.21:259-266,1993)、玉米rab28基因启动子(Busk等,Plant J.11:1285-1295,1997)和玉米Ivr2基因启动子(Pellesehi等,PlantMol.Biol.39:373-380,1999);热诱导型启动子,例如番茄的热番茄hsp80-启动子(美国专利号5,187,267)。
本发明的核酸构建体还优选适当的选择标记和/或复制起点。优选所用的核酸构建体是穿梭载体,它在大肠杆菌(E.coli)能复制(其中构建体包含适当的选择标记和复制起点)并且与细胞增殖同步。按照本发明的构建体可以是例如质粒、杆粒、噬菌粒、粘粒、噬菌体、病毒或人工染色体。
本发明的核酸构建体可用来稳定地或瞬时地转化植物细胞。在稳定的转化中,本发明的外源多核苷酸整合到植物基因组中,因此具有稳定遗传的性状。在瞬时转化中,外源多核苷酸由转化的细胞表达但未整合到基因组中,因此具有瞬时性状。
有多种将外源基因导入单子叶植物或双子叶植物中的方法(Potrykus,I.,Annu.Rev.Plant.Physiol.,Plant.Mol.Biol.(1991)42:205-225;Shimamoto等,Nature(1989)338:274-276)。
使外源DNA稳定整合到植物基因组DNA中的主要方法包括以下两种主要方法:
(i)土壤杆菌介导的基因转移:Klee等(1987)Annu.Rev.PlantPhysiol.38:467-486;Klee和Roger,Cell Culture and Somatic CellGenetics of Plants,第6卷,Molecular Biology of Plant Nuclear Genes,Schell,J.和Vasil,L.K.编著,Academic Publishers,San Diego,Calif.(1989)第2-25页;Gatenby,Plant Biotechnology,Kung,S.和Arntzen,C.J.编著,Butterworth Publishers,Boston,Mass.(1989),第93-112页。
(ii)直接DNA摄入:Paszkowski等,Cell Culture and Somatic CellGenetics of Plants,第6卷,Molecular Biology of Plant Nuclear Genes.Schell,J.和Vasil,L.K.编著,Academic Publishers,San Diego,Calif,(1989),第52-68页;包括原生质体直接摄取DNA的方法,Toriyama,K.等(1988)Bio/Technology 6:1072-1074。植物细胞短时电击诱导的DNA摄入:Zhang等,Plant Cell Rep.(1988)7:379-384。Fromm等,Nature(1986)319:791-793。通过粒子轰击将DNA注入植物细胞或组织中,Klein等,Bio/Technology(1988)6:559-563;McCabe等,Bio/Technology(1988)6:923-926;Sanford,Physiol.Plant.(1990)79:206-209;采用微量加液器***:Neuhaus等,Theor.Appl.Genet.(1987)75:30-36;Neuhaus和Spangenberg,Physiol.Plant.(1990)79:213-217;glass fibers or silicon carbide whisker transformation of cell cultures,embryos or callus tissue(玻璃纤维或金刚砂晶须转化细胞培养物、胚或愈伤组织),美国专利号5,464,765或by the direct incubation of DNAwith germinating pollen(DNA与萌发花粉直接孵育),DeWet等,Experimental Manipulation of Ovule Tissue,Chapman,G.P.和Mantell,S.H.和Daniels,W.编著,Longman,London,(1985),第197-209页;Ohta,Proc.Natl.Acad.Sci.USA(1986)83:715-719。
土壤杆菌***包括采用质粒载体,其含有整合到植物基因组DNA中的确定DNA片段。植物组织的接种方法随植物种和土壤杆菌传递***而变化。广泛应用的方法是叶圆片法,可用任何组织的外植体来进行,它为启动整株植物分化提供良好来源。Horsch等,PlantMolecular Biology Manual A5,Kluwer Academic Publishers,Dordrecht(1988),第l-9页。一种补充的方法是用土壤杆菌传递***与真空渗入相结合。土壤杆菌***在创造转基因双子叶植物方面特别有前途。
有多种直接转移DNA进入植物细胞的方法。电穿孔法,就是将原生质体短时暴露于强电场。微量注射法,就是用极细的微量加液器将DNA直接机械注入细胞内。微粒轰击法,就是使DNA吸附在微粒上例如硫酸镁晶体或钨粒子上,然后用物理方法将微粒加速使其进入细胞或植物组织内。
稳定转化后,进行植物繁殖。最常用的方法是用种子进行植物繁殖。然而由于杂合性,作物缺乏均一性,因此用种子繁殖再生有缺陷,因为植物是按照孟德尔定律控制的遗传变异产生种子。基本上,每粒种子在遗传上都是不同的,每粒种子的生长都会有其独特的性状。因此,优选产生转化植物,致使再生的植物有亲本转基因植物的相同性状和特征。因此,优选用微型繁殖法(micropropagation)再生转化植物,它能快速地、一致地繁殖转化植物。
微型繁殖法是用从所选择的亲本植物或栽培品种切下的一块组织来培育新一代植物的方法。这个方法能大量繁殖具有表达融合蛋白的优选组织的植物,所产生的新一代植物在遗传上与原始植物完全相同,且有其全部特征。微型繁殖法能在短期内大量生产优质的植物材料、快速繁殖所选择的保持原始转基因植物或原始转化植物特征的栽培品种。克隆植物的优点在于植物繁殖的速度,所产生植物的品质一致。
微型繁殖法是多阶段方法,各阶段间需要改变培养基或生长条件。因此,微型繁殖法包括四个基本阶段:第一阶段,初始组织培养;第二阶段,组织培养物扩增;第三阶段,分化和植株形成;第四阶段,温室栽培和锻练。在第一阶段,初始组织培养,建立组织培养物并保证无污染。在第二阶段,初始组织培养物扩增直至组织样本的数量足以满足生产要求。在第三阶段,使第二阶段长出的组织样本***,直到长成单个小植株。在第四阶段,将转化小植株转入温室进行锻练,在此植株对光的耐受性逐渐提高,以便它能在自然环境中生长。
优选如上所述所产生的成熟转化植物根据非生物胁迫耐受性作进一步选择。因此,将转化植物和未转化(野生型)植物暴露在非生物胁迫条件下,例如失水、亚适温度、营养缺乏或优选盐胁迫条件下。盐胁迫可以多种方式实现,例如给植物灌溉高渗溶液、在高渗生长溶液(例如Hoagland溶液)中水培法栽培植物或者在高渗培养基(例如MS培养基)中培育植物。由于不同植物对盐的耐受性极其不同,因此灌溉水、生长溶液或生长培养基中的盐浓度优选按照特定的植物的栽培品种或变种的特殊特征进行调节,以便对植物的生理学和/或形态学施以温和或适度的影响(关于适当浓度的指南请参见Bernstein和Kafkafi,Root Growth Under Salinity Stress:Plant Roots,The HiddenHalf,第三版,Waisel Y,Eshel A和Kafkafi U.(编著)Marcel Dekker Inc.,New York,2002及其中的参考文献)。随后,时时监测暴露于胁迫条件下的植物直至野生型植物出现重大的生理学和/或形态学影响。接着,没有表现出重大生理学和/或形态学影响的转化植物,或者表现出生物量高于野生型的转化植物则被鉴定为非生物胁迫耐受性植物。
虽然现在优选稳定的转化作用,但是叶细胞、分生组织细胞或整株植物的瞬时转化也受本发明重视。
瞬时转化可用上述任何一种直接DNA转移方法,或用修饰植物病毒的病毒感染来实现。
在转化植物宿主中表明有用的病毒包括CaMV、TMV和BV。用植物病毒转化植物在以下文献中有描述:美国专利号4,855,237(BGV)、EP-A 67,553(TMV)、日本公开申请号63-14693(TMV)、EPA194,809(BV)、EPA 278,667(BV);和Gluzman,Y.等,Communicationsin Molecular Biology:Viral Vectors,Cold Spring Harbor Laboratory,New York,第172-189页(1988)。在WO87/0626l中描述了在许多宿主(包括植物)中表达外源DNA所用的假病毒颗粒。
优选本发明的病毒是无毒的因而不能引致严重症状例如降低生长速率、花叶、环斑、卷叶、黄叶、条纹、长痘、长瘤和陷斑。合适的无毒病毒可以是天然存在的无毒病毒或人工减毒的病毒。病毒的减毒可用本领域通晓的方法来实施,包括但不限于亚致死加热、化学处理或定点诱变技术,例如Kurihara和Watanabe(Molecular PlantPathology 4:259-269,2003)、Gal-on等(1992)、Atreya等(1992)和Huet等(1994)所描述的。
合适的病毒株可从现有的资源取得,例如美国典型培养物保藏中心(American Type culture Collection,ATCC)或从感染植物中分离。从感染植物组织中分离病毒可用本领域通晓的技术进行,例如由Foster和Tatlor编著,″Plant Virology Protocols:From Virus Isolation toTransgenic Resistance(Methods in Molecular Biology(Humana Pr),第81卷)″,Humana Press,1998。简而言之,将认为含有高浓度合适病毒的感染植物的组织,优选嫩叶和花瓣,在缓冲液(例如磷酸缓冲液)中研磨获得病毒感染液,其可在后续接种中使用。
植物RNA病毒的构建,将非病毒外源多核苷酸序列导入植物中并使其表达,已为上述和下述文献所证实:Dawson,W.O.等,Virology(1989)172:285-292;Takamatsu等,EMBO J.(1987)6:307-311;French等,Science(1986)231:1294-1297;Takamatsu等,FEBS Letters(1990)269:73-76。
当病毒是DNA病毒时,可对病毒本身进行适当修饰,或者,为了便于构建所需的含有外源DNA的病毒载体,可将病毒首先克隆到细菌质粒上。然后,从质粒切下病毒。如果病毒是DNA病毒,则可将细菌的复制起点与病毒DNA连接,再由细菌复制。该DNA的转录和翻译将产生外壳蛋白,将病毒DNA包入其中。如果病毒是RNA病毒,通常进行cDNA克隆,再***质粒中。然后用该质粒进行所有的构建。然后,经过转录质粒的病毒序列,翻译病毒基因产生外壳蛋白,给病毒RNA包衣壳,产生RNA病毒。
植物RNA病毒的构建,将非病毒外源多核苷酸序列(例如在本发明构建体中所包括的那些序列)导入植物中并使其表达,已为上述文献以及美国专利号5,316,931所证实。
在一个实施方案中,提供植物病毒多核苷酸,其中在病毒多核苷酸中缺失了天然外壳蛋白的编码序列,***了非天然植物病毒外壳蛋白编码序列和非天然启动子、优选非天然外壳蛋白编码序列的亚基因组启动子,其能在植物宿主内表达,包装重组植物病毒多核苷酸,确保宿主被重组植物病毒多核苷酸***感染。另一方面,外壳蛋白基因可以因***其中的非天然多核苷酸序列所失活,因此产生蛋白。重组植物病毒多核苷酸可含有一个或多个另外的非天然亚基因组启动子。每个非天然亚基因组启动子能在植物宿主内转录或表达相邻基因或多核苷酸序列,但不能彼此重组,也不能与天然亚基因组启动子重组。如果包括不止一个多核苷酸序列,那么非天然(外源)多核苷酸序列可以邻近在天然植物病毒亚基因组启动子或天然和非天然植物病毒亚基因组启动子***。在寄主植物中,非天然多核苷酸序列在亚基因组启动子的控制下转录或表达,产生所需产物。
在第二个实施方案中,提供重组植物病毒多核苷酸,如同在第一个实施方案中一样,只是将天然外壳蛋白编码序列置于一个非天然外壳蛋白亚基因组启动子邻近位置而不是置于非天然外壳蛋白编码序列邻近位置。
在第三个实施方案中,提供重组植物病毒多核苷酸,其中天然外壳蛋白基因邻近其亚基因组启动子,在所述病毒多核苷酸中***了一个或多个非天然亚基因组启动子。所***的非天然亚基因组启动子能够在植物宿主内转录或表达相邻基因,但不能彼此重组,也不能与天然亚基因组启动子重组。非天然多核苷酸序列可以邻近在非天然亚基因组植物病毒启动子***,以便在寄主植物中,所述序列在亚基因组启动子的控制下转录或表达,产生所需产物。
在第四个实施方案中,提供重组植物病毒多核苷酸,如同在第三个实施方案中一样,只是天然外壳蛋白编码序列被非天然外壳蛋白编码序列所置换。
病毒载体被由重组植物病毒多核苷酸编码的外壳蛋白包入衣壳内,产生重组植物病毒。重组植物病毒多核苷酸或重组植物病毒常常用来感染合适的寄主植物。重组植物病毒多核苷酸能在宿主内复制、在宿主内***分布、在宿主内转录或表达外源基因(外源多核苷酸),产生所需的蛋白质。
给植物接种病毒的技术可在以下文献中找到:Foster和Taylor编著,″Plant Virology Protocols:From Virus Isolation to TransgenicResistance(Methods in Molecular Biology(Humana Pr),第81卷)″,Humana Press,1998;Maramorosh和Koprowski编著,″Methods inVirology″,第7卷,Academic Press,New York 1967-1984;Hill,S.A.″Methods in Plant Virology″,Blackwell,Oxford,1984;Walkey,D.G.A.″Applied Plant Virology″,Wiley,New York,1985;Kado和Agrawa编著,″Principles and Techniques in Plant Virology″,Van Nostrand-Reinhold,New York。
除上所述,也可以将本发明的多核苷酸引入叶绿体基因组中,从而能使叶绿体表达。
将外源多核苷酸序列引入叶绿体基因组中的技术是已知的。该技术包括下述程序。首先,化学处理植物细胞以减少每个细胞叶绿体的数目至大约一个。然后,用粒子轰击将外源多核苷酸导入细胞中,旨在将至少一个外源多核苷酸分子导入叶绿体中。选择外源多核苷酸以便通过同源重组整合到叶绿体基因组,这由叶绿体所固有的酶容易实现。最后,除了目标基因外,外源多核苷酸还包括至少一段源自叶绿体基因组的多核苷酸序列。此外,外源多核苷酸包括选择标记,其用作通过序贯选择程序确保选择后全部或几乎全部的叶绿体基因组拷贝包括外源多核苷酸。有关该技术更详细的内容参见美国专利号4,945,050和5,693,507,所述专利通过引用结合到本文中。因此,多肽可由叶绿体的蛋白质表达***产生并且可以整合到叶绿体内膜中。
由于植物非生物胁迫耐受性可以包括多个起相加作用或协同作用的基因(参见例如Quesda等,Plant Physiol.130:951-063,2002),本发明也提出在一种寄主植物中表达多种外源多核苷酸,从而获得优良的非生物胁迫耐受性。
在一种寄主植物中表达多种外源多核苷酸,可通过将多个核酸构建体共同导入一个植物细胞中来实现,每个核酸构建体都包括不同的外源多核苷酸。可用本文上述方法将转化细胞再生成为成熟植株。
或者,在一种寄主植物中表达多种外源多核苷酸,也可通过将一个包括多个不同的外源多核苷酸的核酸构建体共同导入一个植物细胞中来实现。可以设计具有一个启动子序列的构建体,该启动子序列能够转录包括所有不同外源多核苷酸序列的多顺反子信使。为了能共同翻译由多顺反子信使编码的不同多肽,多核苷酸序列可以通过内部核糖体进入位点(IRES)序列相互连接,该核糖体进入位点序列促进位于IRES序列下游的多核苷酸序列的翻译。在这种情况下,所转录的编码上述不同多肽的多顺反子RNA分子,自加帽的5’端起开始翻译,而且自多顺反子RNA分子的两个内部IRES序列开始翻译,从而在细胞内产生所有不同的多肽。或者,构建体可以包括几个启动子序列,每个启动子序列都与不同的外源多核苷酸序列连接。
使用上文所述方法,可以将用包括多种不同的外源多核苷酸的构建体转化的植物细胞再生成为成熟植株。
或者,在一种寄主植物中表达多种外源多核苷酸,可以通过将不同的核酸构建体导入多种植物中来实现,其中所述不同的核酸构建体包括不同的外源多核苷酸。然后,可以采用常规植物育种技术,让再生的转化植物杂交,从所产生的子代中选出具有优良的非生物胁迫耐受性和/生物量性状的子代。
因此,本申请提供利用新的非生物胁迫耐受性基因安全、经济有效地提高各种经济作物的非生物胁迫耐受性和/或生物量的方法。
对本领域普通技术人员来说,通过以下非限制性的实施例,本发明的其它目的、优势和新的特征,是显而易见的。另外,本文上述的和在所附权利要求部分中要求保护的本发明的各个不同的实施方案和各个不同的方面,都可以在以下实施例中找到实验支持。
实施例
下面参考以下实施例以及上文的描述,以非限制性的方式说明本发明。
一般而言,本文所用的命名法和本发明中所用的实验室程序包括分子技术、生物化学技术、微生物学技术和重组DNA技术。这些技术在文献中都有详细的解释。参见例如″Molecular Cloning:Alaboratory Manual″Sambrook等(1989);″Current Protocols in MolecularBiology″第I-III卷Ausubel,R.M.编著(1994);Ausubel等,″CurrentProtocols in Molecular Biology″,John Wiley and Sons,Baltimore,Maryland(1989);Perbal,″A Practical Guide to Molecular Cloning″,JohnWiley&Sons,New York(1988);Watson等,″Recombinant DNA″,Scientific American Books,New York;Birren等(编著)″GenomeAnalysis:A Laboratory Manual Series″,第1-4卷,Cold Spring HarborLaboratory Press,New York(1998);美国专利号4,666,828、4,683,202、4,801,531、5,192,659和5,272,057中所述的方法;″Cell Biology:ALaboratory Handbook″,第I-III卷Cellis,J.E.编著(1994);″CurrentProtocols in Immunology″,第I-III卷,Coligan J.E.编著(1994);Stites等(编著),″Basic and Clinical Immunology″(第8版),Appleton&Lange,Norwalk,CT(1994);Mishell和Shiigi(编著),″Selected Methods inCellular Immunology″,W.H.Freeman and Co.,New York(1980);通用的免疫测定在专利文献和科学文献中有全面的描述,参见例如美国专利号3,791,932、3,839,153、3,850,752、3,850,578、3,853,987、3,867,517、3,879,262、3,901,654、3,935,074、3,984,533、3,996,345、4,034,074、4,098,876、4,879,219、5,011,771和5,281,521;″Oligonucleotide Synthesis″Gait,M.J.编著(1984);″Nucleic AcidHybridization″Hames,B.D.和Higgins S.J.编著,(1985);″Transcriptionand Translation″Hames,B.D.和Higgins S.J.编著,(1984);″Animal CellCulture″Freshney,R.I.编著(1986);″Immobilized Cells and Enzymes″IRL Press,(1986);″A Practical Guide to Molecular Cloning″Perbal,B.,(1984);和″Methods in Enzymology″,第1-317卷,Academic Press;″PCR Protocols:A Guide To Methods And Applications″,AcademicPress,San Diego,CA(1990);Marshak等,″Strategies for ProteinPurification and Characterization-A Laboratory Course Manual″CSHLPress(1996);所有这些文献都通过引用全部结合到本文中。通过本文件还可以提供其他的一般性参考文献。相信其中这些程序是本领域众所周知的,只是为方便读者而提供。
除非另有定义,否则本文所用的所有科技术语的含义与本发明所属领域普通技术人员所理解的含义相同。虽然在本发明的实践或检验中可应用与本文描述的类似或等同的方法与材料,但是下文还是说明了合适的方法与材料。
实施例1
鉴定推定非生物胁迫耐受性基因
推定非生物胁迫耐受性(ABST)基因选自番茄表达序列标签(EST)和cDNA的NCBI数据库。数据库序列是用LEADSTM软件(Compugen)聚类和装配的。聚类导致超过20,000个聚类,每一聚类都代表不同的基因。通过汇集包括在含每个聚类的序列记录中的全部关键词,汇编了每个聚类的表达概况。然后筛选包括来源于文库的多核苷酸的聚类,该文库用与ABST有关的关键词确认。将所选聚类进一步过滤,排除任何每个聚类包括超过100EST的聚类和/或任何少于50%序列用ABST相关关键词注释的聚类。
现有技术ABST植物基因可从下述出版物确认:Quesada等(PlantPhysiol.130:951-963,2002);Apse和Blumwald(Curr Opin Biotechnol.13:146-150,2002);Rontein等(Metab Eng 4:49-56,2002);及其中的参考文献。运用BLAST程序,将已知的植物ABST基因与聚类的番茄核酸序列进行比对。e-分值小于5的番茄序列被鉴定为ABST直向同源物。通过使用关键词“root(根)”、“crown gall(冠瘿)”、“nutrient(营养物)”、“callus(愈伤组织)”、“disease(病害)”、“pathogen(病原体)”、“elicitor(激发子)”和“pseudomonas(假单胞杆菌)”,可搜索聚类的番茄序列记录,鉴定出另外的现有技术番茄ABST基因。
最后,将所有经鉴定的现有技术ABST基因与如上所述选择的番茄基因聚类输出组匹配(运用BLAST软件进行序列比对)。结果,在与现有技术ABST基因匹配的聚类输出组中所选出的基因的大约40%被证实是已知的ABST基因,表明所选聚类的其余基因能够潜在提高植物的非生物胁迫耐受性。
运用Vector NTI程序组(suite)(InforMax,U.K.)第6版(HastingSoftware,Inc:
www.generunner.com/),根据ORF(可读框)的存在,对所选出的多核苷酸序列(表1a)进行分析。使用Blast(
www.ncbi.nlm.nih.gov/BLAST/),将每个这样的多核苷酸序列中所鉴定的ORF与Genbank数据库序列进行比较;将表现出与一个或多个Genbank序列有最高同源性的ORF作图,以鉴定ATG起始密码子。然后,将该OPF的ATG起始密码子位置与经鉴定的多核苷酸序列的ATG起始密码子位置进行比较,以证实本文所述的18个序列中的每个序列都包括全长ORF和ATG起始密码子(因而鉴定为“推定ABST基因”)。
表1a
推定ABST基因
ABST No. | SEQ ID No. |
1 | 1 |
3 | 2 |
5 | 3 |
6 | 4 |
10 | 5 |
11 | 6 |
12 | 7 |
19 | 8 |
22 | 9 |
24 | 10 |
26 | 11 |
27 | 12 |
36 | 13 |
37 | 14 |
39T0 | 15 |
39T1 | 16 |
49T0 | 17 |
49T1 | 18 |
运用BLAST软件,从NCBI数据库中鉴定出ABST多肽同源物(表1b)。
表1b
ABST同源物
ABST推定基因SEQ ID No. | ABST多肽同源物NCBI检索号 | ABST多肽同源物SEQ ID No. | 同源性(%) |
1 | BAA96366 | 39 | 98 |
1 | AAS47510 | 40 | 98 |
1 | NP_567151 | 41 | 97 |
1 | NP_567104 | 42 | 96 |
1 | AAK55664 | 43 | 96 |
1 | P46298 | 44 | 97 |
1 | T01338 | 45 | 96 |
1 | T47888 | 46 | 95 |
1 | BAD09465 | 47 | 92 |
1 | Q05761 | 48 | 91 |
1 | BAD09464 | 49 | 88 |
1 | CAA79496 | 50 | 84 |
1 | EAJ94592 | 51 | 85 |
4 | NP_188036 | 52 | 76 |
4 | NP_035977 | 53 | 70 |
4 | XP_342608 | 54 | 69 |
4 | T09295 | 55 | 60 |
4 | NP_564717 | 56 | 59 |
4 | AAM63624 | 57 | 59 |
9 | P37707 | 58 | 93 |
9 | CAD37200 | 59 | 81 |
9 | CAA04664 | 60 | 78 |
9 | AAM64572 | 61 | 77 |
9 | NP_189345 | 62 | 77 |
9 | NP974979 | 63 | 60 |
13 | AAC49992 | 64 | 88 |
13 | T10804 | 65 | 87 |
13 | AAL38357 | 66 | 87 |
13 | NP_188245 | 67 | 87 |
13 | NP_193465 | 68 | 87 |
13 | AAG44945 | 69 | 86 |
13 | T07819 | 70 | 86 |
13 | T12632 | 71 | 86 |
13 | CAC39073 | 72 | 86 |
13 | T01648 | 73 | 86 |
13 | AAF90121 | 74 | 86 |
13 | S48116 | 75 | 86 |
13 | AAO86710 | 76 | 86 |
13 | T14002 | 77 | 85 |
13 | T14001 | 78 | 85 |
13 | T48886 | 79 | 85 |
13 | T14314 | 80 | 85 |
13 | P33560 | 81 | 85 |
13 | P21653 | 82 | 85 |
13 | T14000 | 83 | 85 |
13 | T48884 | 84 | 85 |
13 | P24422 | 85 | 85 |
13 | AAB53329 | 86 | 85 |
14 | NP_200279 | 87 | 67 |
14 | AAM64276 | 88 | 67 |
14 | AAO72577 | 89 | 66 |
14 | NP_175518 | 90 | 64 |
14 | BAC78588 | 91 | 64 |
14 | BAD03011 | 92 | 62 |
实施例2
推定ABS耐受性基因的分离
按照生产商(
www.mrcgene.com/tri.htm)提供的实验方案,用Tri试剂(Molecular Research Center,Inc.),从4周龄番茄根组织和叶组织中提取RNA。按照生产商的说明书,用M-MuLV逆转录酶(RT)(Roche)和T16NN DNA引物,从提取的mRNA中获得互补DNA分子。按照生产商提供的实验方案,用下表2给出的引物和PFU校正DNA聚合酶(Promega-
www.promega.com/pnotes/68/7381 07/738107.html),通过PCR扩增SEQ ID NO:1、4、8-9和12-14中所示的cDNA序列。另外的限制性内切酶位点被加在每一引物的5’引物端,以促进将推定ABS耐受性基因克隆到双元载体中。
表2
用于扩增推定ABS耐受性(ABST)基因的PCR引物
ABST基因SEQ ID No | 正向引物SEQ ID No | 反向引物SEQ ID No | 上游限制位点 | 下游限制位点 |
1 | 21 | 22 | BamHI | SacI |
4 | 23 | 24 | BamHI | SacI |
8 | 25 | 26 | BamHI | SacI |
9 | 27 | 28 | XbaI | SmaI |
12 | 29 | 30 | BamHI | SacI |
13 | 31 | 32 | BamHI | SacI |
14 | 33 | 34 | BamHI | SmaI |
实施例3
克隆推定ABST基因
将所得PCR平端产物,用PCR纯化试剂盒(Qiagen,德国)纯化,用适当的限制酶(Roche)消化,然后***到双元质粒载体pPI中。质粒pPI的构建,是通过将源自pGL3基础质粒载体(Promega,检索号U47295;bp 4658-4811)的合成poly-(A)信号序列***双元载体pBI101.3(Clontech,检索号U12640)的HindII限制位点。和
将所得pPI质粒,用限制酶(BamHI和SacI;MBI Fermentas)消化,用PCR纯化试剂盒(Qiagen,德国)纯化。然后,将开放的pPI构建体与本文上述的七个PCR产物中每一个连接。用含有T4DNA连接酶(Roche)的连接混合物进行连接反应,按照生产商的说明书实施。
用MicroPulser电穿孔器(Biorad)、0.2cm的样品池(Biorad)和EC-2电穿孔程序(Biorad),通过电穿孔法将含有推定ABST基因的pPI构建体导入大肠杆菌(E.coli)DH5感受态细胞中。将经处理的细胞在LB液体培养基中于37℃培养1小时,然后接种到补充卡那霉素(50mg/L;Sigma)的LB琼脂平板上,37℃培养16小时。对选择培养基上显色的菌落,用SEQ ID NO:35-36所示的引物,通过PCR进行分析,将引物设计成跨过pPI质粒的***序列。在1.5%琼脂糖凝胶上分离所得的PCR产物,分离预期大小的DNA片段,用ABI 377测序仪(Amersham Biosciences Inc.)测序,以确认正确的DNA序列被恰当地导入大肠杆菌细胞中。
实施例4
产生包含推定ABST基因和与之有效连接的
植物启动子的双元载体
产生包含花椰菜花叶病毒35S启动子的双元载体:将花椰菜花叶病毒35S启动子序列(如SEQ ID NO:19所示)***上述每个pPI构建体中的推定ABST基因上游。用限制性内切核酸酶HindIII和BamHI(Roche),从pBI121质粒(Clontech,检索号AF485783)中分离启动子。将分离的启动子连接到用相同酶消化的pPI构建体。总共产生了七个ppI构建体,每个构建体都包含位于推定ABST基因上游的CaMV 35S启动子,该ABST基因具有SEQ ID NO:1、4、8、9、12、13或14中所示的序列。
产生包含At6669启动子的双元载体:将At6669启动子序列(如SEQ ID NO:20所示)***上述每个pPI双元构建体中的推定ABST基因上游。用SEQ ID NO:37-38中所示的引物,通过PCR扩增,从拟南芥变种Co10基因组DNA中分离启动子。将PCR产物纯化(Qiagen,德国),再用限制性内切核酸酶HindIII和BamHI(Roche)消化。将所得启动子序列导入用相同酶消化的开放双元构建体。总共产生了七个pPI构建体,每个构建体都包含位于推定ABST基因上游的At6669启动子,该ABST基因具有SEQ ID NO:1、4、8、9、12、13或14中所示的序列。
实施例5
确认转基因拟南芥中At6669启动子的活性
对At-6669启动子调节植物内pPI载体所携带基因的转录的能力进行检验。因此,将At6669启动子***pPI双元载体的萤光素酶报道基因上游。用下面实施例6描述的方法,将双元载体导入拟南芥植物中。用Meissner等(Plant J.22:265,2000)描述的方法,用超低光检测照相机(Princeton Instruments Inc.,USA),测试成熟转化T2拟南芥植物在暗室内的生物发光。在转化植物的花和根分生组织内观察到显示阳性萤光素酶活性的发光(图2)。
实施例6
用含有推定ABST基因的双元载体转化根瘤土壤杆菌细胞
用以上实施例4描述的每个双元载体转化土壤杆菌细胞。将两个附加的双元构建体,其萤光素酶报道基因代替ABST基因(置于35S或At6669启动子的下游),用作阴性对照。
通过电穿孔,将双元载体导入根瘤土壤杆菌GV301或LB4404感受态细胞(约109细胞/m1)中。用MicroPulser电穿孔器(Biorad)、0.2cm样品池(Biorad)和EC-2电穿孔程序(Biorad)实施电穿孔。将经处理的细胞用LB液体培养基于28℃培养3小时,然后接种到补充庆大霉素(50mg/L;用于土壤杆菌菌株GV301)或链霉素(300mg/L;用于土壤杆菌菌株LB4404)和卡那霉素(50mg/L)的LB琼脂平板上,于28℃培养48小时。对选择培养基上显色的土壤杆菌菌落,用SEQ ID NO:35-36中所示的引物,通过PCR进行分析,将PCR设计成跨过pPI质粒中的***序列。如以上实施例4中所述,对所得的PCR产物进行分离和测序,以确认正确的ABST序列被恰当地导入土壤杆菌细胞中。
实施例7
用推定ABST基因转化拟南芥植物
用Clough和Bent(10)和Desfeux等(11)所描述的Floral Dip程序,稍作修改,转化拟南芥Columbia植物(T0植物)。简而言之,将T0植物种在250ml花盆中,花盆中装满以湿泥炭为基础的生长混合物。将这些花盆用铝箔和塑料圆顶盖住,于4℃保持3-4天,然后揭盖,于18-24℃,经16/8小时光/暗周期,在生长室中培养。T0植物在开花前6天就可用于转化。
将上述实施例6所产生的携带双元载体的单个土壤杆菌菌落,在补充卡那霉素(50mg/ml)和庆大霉素(50mg/ml)的LB培养基中进行培养。将这些培养物于28℃剧烈振荡下培养48小时,然后以4000rpm离心5分钟。将包含土壤杆菌细胞的沉淀重悬浮于含有下述成分的转化培养基中:半浓度(2.15g/L)Murashig-Skoog(Duchefa)、0.044μM苄氨基嘌呤(Sigma)、112μg/L B5 Gambourg维生素(Sigma)、5%蔗糖和用双蒸水制备的0.2ml/L Silwet L-77(OSI Specialists,CT),pH5.7。
T0植物的转化,是通过把每株植物倒放在土壤杆菌悬液中进行,使得地上的植物组织浸泡3-5秒。随即将每个已接种的T0植物放在塑料盘内,然后盖上干净的塑料圆顶以保持湿度,置暗处室温18小时,以促进感染和转化。然后将已转化的(转基因)植物揭开盖子并转入温室使其恢复和成熟。转基因T0植物在温室中生长3-5周直至长角果呈褐色和干燥。从植物收获种子,置室温直至播种。
为了产生含基因的T1和T2转基因植物,因此将从转基因T0植物收集的种子先浸入70%乙醇1分钟,随后浸入5%次氯酸钠和0.05%triton 5分钟,进行表面消毒。将经表面消毒的种子用无菌蒸馏水彻底清洗,然后放在培养板上,该培养板装有半强度的Murashig-Skoog(Duchefa)、2%蔗糖、0.8%植物琼脂、50mM卡那霉素和200mM羧苄青霉素(Duchefa)。将培养板于4℃保温48小时,然后转入生长室于25℃再培养一周。将活的T1拟南芥植物转入新鲜培养板中,再培养一周。培养后,将T1植物从培养板移栽到含有生长混合物的250ml花盆中。让转基因植物在温室里长至成熟。在与培养和生长T1植物相同的条件下,将从T1植物收获的种子播种并让其生长至成熟作为T2植物。
实施例8
评估在非生物胁迫条件下栽培的转基因植物的生长方法:
将上述产生的T1或T2转基因植物逐一移栽到花盆中,花盆中装有泥炭和蛭石(容积比分别为3∶2)的生长混合物。盖住花盆锻练24小时,然后以完全随机的次序置于温室,用自来水灌溉7天(每3-5天从盆底供水)。此后,一半植物灌溉盐溶液(100mM NaCl和5mM CaCl2)以诱导盐胁迫(胁迫条件)。另一半植物继续灌溉自来水(正常条件)。全部植物在100%RH下温室生长28天,然后收获(地上组织),并称重(立即或烘箱50℃干燥24小时后)。
结果:
用3种不同的ABST基因转化的T1植物和用萤光素酶报道基因转化的植物,生长在正常或胁迫条件下,未观察到两者间植物鲜重的显著性差异(图3和下表3)。用At6669启动子调控下的SEQ ID NO:1转化的T1植物,在处于胁迫条件下时,保持有它们鲜重的71%,而对照植物(携带处于At6669启动子调控下的萤光素酶基因),在相似的胁迫条件下,仅有它们鲜重的61%。
表3
用水或盐溶液灌溉的T1转基因拟南芥植物的鲜重
转基因(SEQ IQ NO) | 启动子 | N行1 | 灌溉溶液(mMNaCl) | 平均值(g) | 标准误 |
萤光素酶 | At6669 | 2 | 0 | 0.7925 | 0.0275 |
萤光素酶 | At6669 | 2 | 100 | 0.485 | 0.045 |
13 | At6669 | 8 | 0 | 0.81625 | 0.020305 |
13 | At6669 | 8 | 100 | 0.4725 | 0.029246 |
1 | At6669 | 8 | 0 | 0.7875 | 0.026032 |
1 | At6669 | 8 | 100 | 0.55875 | 0.044699 |
8 | At6669 | 8 | 0 | 0.8575 | 0.023088 |
8 | At6669 | 8 | 100 | 0.440625 | 0.011198 |
1N行表示所测的独立转化事件的植株数。对于每个转基因,用3-5个独立的转化事件,每个转化事件用1-3个植株。
用处于35S启动子调控下的SEQ ID NO:7或14转化的T2植物所累积的生物量显著高于对照植物,与生长条件无关。如图4A和下表4所示,用SEQ ID NO:7和14转化的植物,生长在胁迫条件下的平均鲜重,比生长在相似胁迫条件下的对照植物的平均鲜重分别高15%和24%。同样,用SEQ ID NO:7或14转化的植物,生长在正常条件下的平均鲜重,比生长在相似正常条件下的对照植物的平均鲜重分别高21%和27%。
从处于35S启动子调控下的SEQ ID NO:4转化的T2植物中也观察到类似现象。因此,如下面图4A和表4所示,用SEQ ID NO:4转化的植物的平均鲜重,比生长在胁迫和正常条件下的对照植物的平均鲜重分别高14%和7%,同样,用处于At6669启动子调控下的SEQ ID NO:4转化的T2植物,比生长在胁迫和正常条件下的对照植物的生物量分别高1.3和5%。然而,在实验条件下,这些差异在统计学上没有什么不同。
表4
用水或盐溶液灌溉的T2转基因拟南芥植物的鲜重
转基因(SEQ IQ NO) | 启动子 | 1N行 | 灌溉溶液(mM NaCl) | 平均值(g) | 标准误 |
萤光素酶 | CaMV-35S | 11 | 0 | 0.352727 | 0.011208 |
萤光素酶 | CaMV-35S | 11 | 100 | 0.280909 | 0.010484 |
9 | CaMV-35S | 11 | 0 | 0.426364 | 0.019599 |
9 | CaMV-35S | 11 | 100 | 0.322727 | 0.027306 |
12 | GaMV-35S | 11 | 0 | 0.374545 | 0.015746 |
12 | CaMV-35S | 11 | 100 | 0.249091 | 0.020647 |
1 | CaMV-35S | 8 | 0 | 0.36625 | 0.034171 |
1 | CaMV-35S | 8 | 100 | 0.265 | 0.031225 |
13 | CaMV-35S | 11 | 0 | 0.349091 | 0.013515 |
13 | CaMV-35S | 11 | 100 | 0.293636 | 0.019921 |
14 | CaMV-35S | 11 | 0 | 0.446364 | 0.025558 |
14 | CaMV-35S | 11 | 100 | 0.348182 | 0.023772 |
8 | CaMV-35S | 11 | 0 | 0.310909 | 0.015223 |
8 | CaMV-35S | 11 | 100 | 0.253636 | 0.01539 |
4 | CaMV-35S | 11 | 0 | 0.379091 | 0.010992 |
4 | CaMV-35S | 11 | 100 | 0.318182 | 0.013336 |
1N行代表所测的独立转化事件的植株数。对于每个转基因,用3-
5个独立的转化事件,每个转化事件用1-3个植株。
用处于At6669启动子调控下的SEQ ID NO:1和13转化的,并生长在胁迫条件下的T2植物,比生长在类似胁迫条件下的对照植物的生物量高出许多。用处于At6669启动子调控下的SEQ ID NO:1和13转化的,并生长在胁迫条件下的T2植物的平均鲜重,比生长在类似条件下的对照植物的平均鲜重分别高37%和21%(图4B和下表5)。当这些转基因植物(携带处于At6669启动子调控下的SEQ ID NO:1和13)生长在正常条件下时,未发现其生物量显著高于对照植物。
表5
用水或盐溶液灌溉的T2转基因拟南芥植物的鲜重
转基因(SEQ IQ NO) | 启动子 | 1N行 | 灌溉溶液(mM NaCl) | 平均值(g) | 标准误 |
萤光素酶 | At6669 | 6 | 0 | 0.3 | 0.010328 |
萤光素酶 | At6669 | 6 | 100 | 0.125 | 0.009916 |
13 | At6669 | 6 | 0 | 0.286667 | 0.024449 |
13 | At6669 | 6 | 100 | 0.151667 | 0.007032 |
1 | At6669 | 6 | 0 | 0.305 | 0.03423 |
1 | At6669 | 6 | 100 | 0.171667 | 0.012225 |
4 | At6669 | 6 | 0 | 0.315 | 0.049983 |
4 | At6669 | 6 | 100 | 0.126667 | 0.005578 |
12 | At6669 | 6 | 0 | 0.263333 | 0.012824 |
12 | At6669 | 6 | 100 | 0.098333 | 0.007923 |
8 | At6669 | 6 | 0 | 0.228333 | 0.020235 |
8 | At6669 | 6 | 100 | 0.121667 | 0.004014 |
1N代表所测的独立转化事件的植株数。对于每个转基因,用3-5个独
立的转化事件,每个转化事件用1-3个植株。
这些结果表明,所分离的推定ABST基因,如SEQ ID NO:1和13所示,能够提高植物对非生物胁迫例如盐胁迫的耐受性。此外,所分离的SEQ ID NO:7、14(还可能是4)中所示的推定ABST基因,能够显著地提高生长在胁迫和正常条件下的植物的生物量。
因此,结果清楚地表明,本文所述的推定ABST基因易于分离,并能用以显著地提高植物对非生物胁迫的耐受性和/或生物量。
可以理解的是,本发明的某些特征在各个实施方案里已作清楚、完整说明,也可以结合在一个实施方案里提供。反过来说,本发明的不同特征在一个实施方案里已作说明,为了简明也可以单独地或以任一种适当的再次结合提供。
虽然本发明已结合具体的实施方案予以说明,但是显然许多供选择的方案、修改和变动对于本领域的技术人员来说是显而易见的。因此,所有的这些供选择的方案、修改和变动都落入所附权利要求的精神和范围内。本发明说明书中提及的所有出版物、专利、专利申请和根据其检索号所标识的序列在本文中通过引用全部结合到本发明说明书中,其程度如同每个出版物、专利或专利申请或根据其检索号所标识的序列具体而单独地指明通过引用结合到本文中一样。此外,本申请中的任何参考文献的引用或确认都不得解释为承认这些参考文献是可作为本发明的现有技术获取的。
引用的参考文献
(附加参考文献是在上文中引用的)
1.www.fao.org/ag/agl/agll/spush/degrad.htm.
2.www.fao.org/ag/agl/aglw/watermanagement/introduc.stm
3.McCue KF,Hanson AD(1990).Drought and salt tolerance:towards understandingand application.Trends Biotechnol 8:358-362.
4.Flowers TJ,Yeo Ar(1995).Breeding for salinity resistance in crop plants:wherenext?Aust J Plant Physiol 22:875-884.
5.Nguyen BD,Brar DS,Bui BC,Nguyen TV,Pham LN,Nguyen HT(2003).Identification and mapping of the QTL for aluminum tolerance introgressed fromthe new source,ORYZA RUFIPOGON Griff.,into indica rice(Oryza sativa L.).Thcor Appl Gcnet.106:583-93.
6.Sanchez AC,Subudhi PK,Rosenow DT,Nguyen HT(2002).Mapping QTLsassociated with drought resistance in sorghum(Sorghum bicolor L.Moench).Plant Mol Biol.48:713-26.
7.Quesada V,Garcia-Martinez S,Piqueras P,Ponce MR,Micol JL(2002).Geneticarchitecture of NaCl tolerance in Arabidopsis.
Plant Physiol.130:951-963.
8.Apse MP,Blumwald E(2002).Engineering salt tolerance in plants.Curr OpinBiotechnol.13:146-150.
9.Rontein D,Basset G,Hanson AD(2002).Metabolic engineering ofosmoprotectant accumulation in plants.
Metab Eng 4:49-56
10.Clough SJ,Bent AF(1998).Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.Plant J 16:735-43
11.Desfeux C,Clough SJ,Bent AF(2000).Female reproductive tissues are theprimary target of Agrobacterium-mediated transformation by the Arabidopsisfloral-dip method.Plant Physiol 123:895-904.
序列表
<110>伊沃基因有限公司(Evogene Ltd.)
<120>提高植物非生物胁迫耐受性和/或生物量的方法
<130>CPCH0564181P
<160>92
<170>PatentIn version 3.2
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<213>番茄(Lycopersicon esculentum)
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atgggtcgta tgcacagtcg tggtaagggt atttcagctt ctgctctccc ttacaagaga 60
actcctccta gttggctcaa gatctctgct ccagatgttg aggacaacat ctgcaagttc 120
gctaagaaag gattgacccc ttcacagatt ggtgtgattc ttcgtgattc tcatggaatt 180
gcacaagtga agagtgttac tggtagcaag atcttgcgta tcctcaaggc acatgggctt 240
gcacctgaga ttccagagga tttgtaccac ctgattaaga aggctgttgc cattaggaag 300
catttggaga ggaacaggaa ggataaggat tctaagttcc gtttgatttt ggtggagagc 360
aggattcatc gccttgctcg ttattacaag aaaacaaaaa agctcccacc tgtctggaaa 420
tacgaatcta ccactgctag cacacttgtg gcatag 456
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<212>DNA
<213>番茄(Lycopersicon esculentum)
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atggctattc ctcttcaatt ctctagtata tccactcgca cagatctctc cttgccggag 60
actagaactt tcaggttacc gaaacctttc tccgtcatca gatgctccgc cggcgaacct 120
gttccttcct cgtcggctac tgctgagtca gagttcgatg ctaaggtttt ccggaagaat 180
ctgaccagaa gtgctaatta caatcgtaaa ggttttggac acaaagaagc tactcttgaa 240
ctcatgaatc gcgaatatac cagtgatatc atcaagaaat tgaaggagaa tgaatttgag 300
tatacatggg gaaacgtaac cgtaaaactt gcagagtcct atggtttctg ttggggggtt 360
gagcgtgcag ttcagattgc ttatgaagcg aggaaacagt ttccaacaga gaggatttgg 420
ataactaatg aaattattca caaccccact gtgaataaga ggctagagga tatggatgtt 480
aagaacattc cacttgagga agggaagaaa aactttgatg ttgttgacaa ggatgatgtt 540
gtggttttgc ctgcttttgg ggctgctgtt gatgaaatgt tggttttgag tgataaaaac 600
gtacaaattg ttgatacaac ctgcccgtgg gtgactaagg tttggaacac ggttgaaaag 660
cacaagaagg gagaatatac ctccattatc catggtaaat atgctcatga ggaaactgtt 720
gcgactgcat cctttgctgg gaaatacatc attgtgaaga acatggcaga ggcaacttat 780
gtctgtgatt atattcttgg aggtaaactt gatggttcta gctcaaccaa agaggcattt 840
atgcagaaat ttaaatatgc agtttctgaa gggtttgatc cggatgttga ccttgtaaaa 900
gctggtattg caaaccaaac aactatgttg aagggagaaa cagaagatat tgggaagttg 960
gtcgagagga ccatgatgca aaaatatggg gtggaaaatg ttaacaacca cttcgtaagt 1020
ttcaacacta tatgcgatgc cacacaagag cgtcaagatg caatgtataa gctggttgag 1080
caaaagctgg atcttatgtt agtgattggt ggctggaact caagtaacac ttcacatcta 1140
caggagattg cagaggaacg tggaattccc tcatactgga ttgacagtga acagagagta 1200
ggtcctggaa acaaaataag ttacaagtta atgcatggtg agttggttga gaaagagaac 1260
ttcttaccgg agggtcctat tacagttggg gtgacatctg gtgcatccac ccccgataag 1320
gttgttgaag atgtccttat caaggtgttt gatatcaagc gcgaggaagc cttacaattg 1380
gcctaa 1386
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<212>DNA
<213>番茄(Lycopersicon esculentum)
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atggctcaac gtgttctaac tcgtgttcac agtcttcgtg aacgtcttga tgctactttg 60
gatgctcatc gcaatgaaat tttgctcttt ctttcaagga tcgaaagcca cgggaaaggg 120
atcttgaaac ctcaccagct actggctgag tttgaatcaa ttcagaaaga agacaaagac 180
aaactgaatg atcatgcctt tgaagaagtc ctgaaatcca ctcaggaagc aattgttttg 240
cccccatggg ttgcacttgc tattcgtttg aggcccggtg tgtgggaata tgtccgtgtg 300
aatgttaatg ctcttagtgt tgaggagctg actgtgcctg agtttttgca attcaaggaa 360
gaacttgtta acggaacttc cagtgataac tttgttcttg aattggattt tgagcccttc 420
actgcatcat ttccaaaacc aaccctcacg aaatcaattg gaaatggagt tgaattcctc 480
aacaggcacc tctctgctaa aatgttccat gacaaggaaa gcatgacccc tcttctcgag 540
tttcttcgag ttcaccacta caatggaaag tcaatgatgc tgaatgatag aattcagaat 600
ttgtatactc tccaaaaagt cctgaggaag gccgaggaat acctcaccac cctttcgcca 660
gaaacttcat actcctcatt tgagcacaag ttccaagaaa ttggcttgga gagaggttgg 720
ggtgacaccg cagagcgtgt tctagagatg atctgcatgc tcctggatct ccttgaggct 780
cctgactcat gtactcttga gaagttcctt agtagaattc ctatggtttt caatgtagtt 840
atactttcac ctcatggata tttcgcccag gaaaatgtct tgggttaccc cgacactggt 900
ggtcaggttg tctatatttt ggatcaagtt cctgccttgg agcgtgagat gctcaagcgc 960
ataaaggagc aaggacttga tatcaaaccg cgtattctta ttgttactcg gcttctccct 1020
gatgcagttg gtaccacttg tggtcagcga ctcgagaagg tatttggaac tgagcattca 1080
catattctta gggtcccctt taggactgaa aagggcattg ttcgcaaatg gatctctcgt 1140
tttgaagtct ggccatacat ggagactttc attgaggatg tggggaaaga aataaccgca 1200
gaactgcaag ctaagccaga tcttattatt ggaaactata gtgagggaaa ccttgcagcc 1260
tccttgttgg ctcacaagtt aggtgtaaca cagtgcacca ttgctcatgc attggagaaa 1320
accaaatatc ctgattctga catttacttg aacaaatttg acgagaaata ccacttctca 1380
gctcagttca cagctgatct tatagcaatg aatcatactg atttcattat caccagcacc 1440
ttccaggaga tagcaggaag caaggacact gttggacagt atgagagcca catggccttc 1500
acaatgcctg gattgtatag agttgttcat ggcattgatg tgttcgaccc caaattcaac 1560
attgtgtcac caggagctga tgtgaatctc tatttcccat actccgaaaa ggaaaagaga 1620
ttgacaactt ttcaccctga aattgaagac ttgctgttta gcgatgttga gaacgaagaa 1680
cacctgtgtg tgttgaagga caggaataag cccatcatat tcaccatggc aagattggac 1740
cgagtgaaga acttaactgg acttgtcgag tggtatgcta agaatccacg actaagggag 1800
ttggttaacc ttgtagtggt tggtggagac cgaagaaagg aatccaaaga cttggaagag 1860
caggcagaga tgaagaagat gtatgaactt ataaagactc acaatttgaa tggccagttc 1920
cgatggattt cttcccagat gaaccgcgtg aggaatgggg aactctacag gtacattgct 1980
gacacaaggg gagctttcgt gcagcctgca ttctacgagg ctttcggtct gactgttgtt 2040
gaggccatga gctgcggttt gcctacattt gcaactaatc aaggtggtcc agctgagatc 2100
atcgttcatg gaaagtctgg tttccaaatt gatccatacc atggcgagca ggctgctgat 2160
ctcctcgctg agttcttcga gaaatgtaag gtagaccctt cacattggga agccatttcc 2220
aagggtggcc ttaagcgtat acaggagaag tacacatggc aaatctactc cgaccggctg 2280
ttgacactag ctgctgttta cgggttctgg aagcacgttt ccaagcttga tcgtcttgaa 2340
attcgtcgtt atcttgagat gttttacgct ctcaaattcc gcaagctggc tgaacttgtc 2400
ccattggctg ttgagtaa 2418
<210>4
<211>522
<212>DNA
<213>番茄(Lycopersicon esculentum)
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atggaagaca aaagcaatga ttattatgca gttttggggt tgaagaagga atgcactgac 60
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gcatcgggga atttgaagtt tgtagatgaa gcaaagaagc aatttcaggc aattcaagaa 180
gcatattctg tgttatcgga tgcaaacaaa aagtttttgt acgatgtagg agtttatgac 240
tctggtgatg atgacgacga aaatggcatg ggtgatttcc tgaatgaaat ggcagctatg 300
atgagccaaa ataagtccaa tgaaaatcag ggagaagaaa cctttgagga attgcaggat 360
atgtttaatg aaatgttcaa cagtgataat ggaacgtttt cttcttcttc ttcttcttct 420
tcttcttctt ggactggaac tccttcaatg tgctctacta catcatctac atcttcaagt 480
gagacttttt taacctttcc ccaacaagag aagttcaggt ga 522
<210>5
<211>1434
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>5
atggctactg ctactactct ctctcctgct gatgctgaaa agctcaacaa cctcaaatct 60
gccgtcgccg gtctaaatca aatcagtgaa aatgagaaat ctggatttat taaccttgtc 120
ggtcgctatc taagtggtga agcacaacac attgactgga gtaagatcca gacgccaact 180
gatgaagttg tggtgcctta tgacaagtta gcacctcttt ctgaagatcc cgcggaaact 240
aagaagcttt tggacaaact tgttgtcctg aagctcaatg gaggcttggg aacaacaatg 300
ggatgcacgg gtcccaaatc agttattgaa gttcgtaatg gtttgacatt ccttgacttg 360
attgtcaagc aaattgaggc actcaatgcc aagttcggat gcagtgttcc cctgcttttg 420
atgaattcgt tcaacaccca cgatgataca ctgaagattg ttgaaaaata tgcaaactca 480
aacattgata ttcatacatt caatcagagc cagtaccctc gcctggttac tgaagacttt 540
gccccacttc catgcaaagg caattccgga aaagatggat ggtaccctcc aggtcatggt 600
gatgttttcc cttctttgat gaatagtgga aagcttgatg cactactagc aaagggcaag 660
gaatatgtct ttgttgcaaa ctctgataat ttgggcgcca ttgttgattt gaaaatctta 720
aatcatttga tcctaaacaa aaatgagtac tgcatggagg ttactcccaa aactttagct 780
gatgtcaaag gtggcacctt aatctcatat gaaggaaaag tacagctatt ggaaatagca 840
caagtccctg atgaacatgt caatgaattc aagtcaattg aaaaattcaa aattttcaac 900
accaacaact tgtgggtgag tcttagtgct attaaaagac ttgtagaagc agatgcactc 960
aagatggaga ttattcccaa cccaaaggaa gtagacggag ttaaagttct tcaacttgaa 1020
actgctgccg gtgctgcgat taagtttttc gaccgggcaa ttggtgctaa tgttcctcga 1080
tctcgtttcc ttcccgtgaa agcaacttca gatttgctcc ttgttcagtc tgatctttac 1140
accttgactg atgagggcta tgtcatccga aacccggcca ggtctaatcc gtccaaccca 1200
tccatcgagt taggacctga attcaagaag gtggccaact tcttaggccg tttcaagtcc 1260
attcccagca tcattgatct aggtagcttg aaggtgaccg gtgatgtatg gtttggatct 1320
agcgttaccc taaaggggaa agtgactgtt gcagccaaat ccggagtgaa gctagaaatt 1380
ccagatggtg ctgtgattgc aaacaaggac atcaatggac ctgaggatat atag 1434
<210>6
<211>1485
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>6
atggcaaaga gtggcatttt ggtaattgtt tcagctcttg ttgttcttgc agtttgtggt 60
gtttttgctg aggagaacga atatgtgttg actttggacc attctaacct cactgagact 120
gttgctaagc acaacttcat tgttgttgaa ttctatgcac cttggtgtgg acactgtaag 180
agtcttgctc ctgagtatga aaaagctgcc tcagagctga gtagtcatga ccctccaatt 240
gttctagcta agtatgatgc aaatgatgaa gccaatagag aactttcaaa acagtacgag 300
atccagggtt tcccaactat taagatattg agagatggag gaaagaaagt tcaagactat 360
aacggtcctc gtgaagcagc tggtattgta tcctacttga agaaacaagt gggtcctgca 420
tctgctgaaa tcaagtcgaa ggaagatgcc acaaacctta ttgatgagaa aagtatcttt 480
gttgttggta tatttccaga cccctccgga gagaaattcg agaactattt aacgctagct 540
gaaaaactgc gaggcgagtt cgattttgct cacactgttg atgctaaaca cctccctcgg 600
ggtggaccag tcaacaagcc cactcttcgt cttctaaagc catttgatga actctttgtt 660
gattttgagg actttgatgt cgatgcaatg gagaagttca tctcagaatc tagtattcct 720
gttgttacta tttttgacaa tgacccaaac aaccatcctt atgttaacaa gttcttcgaa 780
ggcaccaacg ccaaggcatt gctatttgtg aactttagct ctgaatttga tgcttttaag 840
tccaagtaca acgatgttgc tgtgatttac aaaggggatg gggtgagctt tctcttgggt 900
gatgttgagg ctggtcaagg tgcttttgag tacttcggac tgaagccgga acaggcacct 960
gtgatcatca taatggacgc tgatgaacaa aagtatatta aggaccatgt ggaacctgat 1020
gccattgctg cttacttgaa ggattacaag gaaggaaaac tgaagccaca tgtgaagtca 1080
gagcccatcc ctgaagtcaa tgacgaacct gttaaggtgg ttgttaggga taccctccag 1140
gatatggttt acaaatcggg aaaaaatgtg ctgttagagt tctatgcacc ttggtgtggc 1200
cactgcaaga gtctggctcc aattttggat gaagtggctg tatcatttga aagcgatcct 1260
gatgttctca ttgcaaaact ggacgcaacc gcaaatgatc tcccgaaagg tgactttgat 1320
gttcagggat tccctactat gtacttcaga tccgcctctg gtaacttgtc acagtacaat 1380
ggtgagagaa caaaagaggc tatcatcgaa ttcatcgaga agaatcgtgg caagcctgct 1440
cagtcagact ctgccaaagt cgattcagca aaggatgaac tttag 1485
<210>7
<211>250
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>7
atgccgggtg cgctcgcatg gcaggtcaca aaacctaata aagatgcgat ttttgtgttc 60
ggaggggaga tggtacgggg ttttttgccc gactctcctc tctgtgtctg tctttggcct 120
cctttctttt ctagaactgt ctgcaagatc tattcagata aacgtctcat caaatttggc 180
gttaatgtcc ccgaacgatg gtttcgatat ccgcaaagtt atagcagcgc tctacctttt 240
caatggatag 250
<210>8
<211>717
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>8
atgaattctc agatttgcag atctgctaca agagcagcta agtcactcct ttctgcttca 60
tctaagcaga cttctcgtgc tttttcagga ggacgagcag cagctgcagc agccacagtt 120
tcattgagag gagtggtgcc ttctctagcc tcatatggca ggaatgaatc tggaaatgca 180
tctagagctt ggatttctgg tgtgcttgcc cttcctgcag cagcttacat gctccaggag 240
caagaagcac atgctgccga gatggagcgc acctttattg ccatcaagcc agatggagta 300
cagagaggcc tgatttcaga aatcgtatca cggtttgagc gcaagggctt caagctggtt 360
gcaatcaaag ttgtgattcc ttccaaggaa tttgcaaaga agcactatca tgacttgagt 420
gagcgaccat tctttaacgg cttgtgcgac ttccttagct ctggccctgt cttagcaatg 480
gtttgggaag gtgaaggtgt aatcagatat ggaaggaagc ttatcggagc caccgatcca 540
cagaaatctg aacctggaac catcagaggc gatttagctg ttgtagtagg aaggaacatc 600
atccatggca gcgatggccc cgagaccgca aaggatgaga tcaacctatg gtttaaacca 660
gaggagttgg ttaattacac cagcaactct gagaagtggc tatatggtga taactaa 717
<210>9
<211>897
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>9
atggagaata tgcagagcta ttggcaattt ggcgacgagc ttcgaggaca atcaaaagcc 60
tcagaggatc ataaatggtc aacagctgct ataaaattat ctgaacagat gaagtacaaa 120
ggtgaacgta ggaataacct tgacctttca aagagctctg ctgaaattag gcccaggggt 180
aatcatatgt ttcaggaaga taacaagtgg gaaagcctta acttcaatat gttaaatttg 240
gaaagcaaga tgactgaaaa tatgagcaag aatcgcatta tggatagcat tttcaatgca 300
aatccagttt atcttaagcc caattttaac agcttgggaa attcatcttt aagcaagttc 360
aatgctagca actataccaa ggaacctagc aagaataaca ataacaacgt tgagagcaca 420
aatggaaata actccgttga caaaaggttt aagactctgc ctgctgctga aacactgccg 480
aagaatgagg ttcttggtgg atatatattt gtttgtaaca atgacacaat gcaggaagac 540
ctaaagcgcc tgctctttgg ccttcctcct agatacagag attccgtgag ggcaataaca 600
ccagggttgc ccttgttcct atataattac actactcacc agttgcatgg tatctttgag 660
gcatcgagtt ttggaggttc caacattgat ccaactgcct gggaggataa aaagtgtaaa 720
ggagagtcaa ggttccctgc tcaggtgagg atccgtgtcc ggaaagtctg taatcctttg 780
gaggaagatg ctttcagacc agttttacat cattatgatg gccccaagtt ccgtctggag 840
ctctccattc ctgagacttt ggacttacta gatctctgtg aaaaagccgg tgtgtag 897
<210>10
<211>489
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>10
atgtcacacc ccaccgcagc tccgctatca ggaccacgcc tctcacaacc ctcgtcgtcg 60
gcgatgtctc ccttgtacaa acagaaatct tggtcaccgg acacgtttcg cgacgaggcg 120
tggcagcggc ggaagggtac ccacggaagc tgcctcaaac ggcggagcaa gagcgttacc 180
gatgaggact ttgatgagat taaggcctgt atcgaattag ggtttggatt tgattcgcca 240
gaaatggatc agcgattgtc tgatactttt ccggcgtatg acctgtttta cgccgtgaat 300
aaacaataca ccgacactct ttcaaagact tcctctgtat catcggtcat ctccaattgc 360
gagtcaaccc ttcctcccgt cagtccccac accattgtct ttccaggaga taatccacag 420
gcagtgaaga caaggttgcg gcaatgggca caggtggttg cgtgtgtggt gcgtcaatct 480
tcgtattaa 489
<210>11
<211>489
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>11
atgtcacacc ccaccgcagc tccgctatca ggaccacgcc tctcacaacc ctcgtcgtcg 60
gcgatgtctc ccttgtacaa acagaaatct tggtcaccgg acacgtttcg cgacgaggcg 120
tggcagcggc ggaagggtac ccacggaagc tgcctcaaac ggcggagcaa gagcgttacc 180
gatgaggact ttgatgagat taaggcctgt atcgaattag ggtttggatt tgattcgcca 240
gaaatggatc agcgattgtc tgatactttt ccggcgtatg acctgtttta cgccgtgaat 300
aaacaataca ccgacactct ttcaaagact tcctctgtat catcggtcat ctccaattgc 360
gagtcaaccc ttcctcccgt cagtccccac accattgtct ttccaggaga taatccacag 420
gcagtgaaga caaggttgcg gcaatgggca caggtggttg cgtgtgtggt gcgtcaatct 480
tcgtattaa 489
<210>12
<211>606
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>12
atgggatact ggaaagcaaa ggttcttcca aagatcaagc agatctttga taaaaatgga 60
cccaagaaaa ctgctgctgc tgaggcatgc aagacttttg atcaagctaa ggaggaatat 120
agcaaggagt ttgaagagaa gaagactgag cttcaaccca aagttgttga aatttatgaa 180
gctgctgcag ttgagatcaa gagcttagtg aaggaaccaa agggtgcagg gctgaagaaa 240
aactcagatg gggttcagaa attccttgat gaccttgtca agattgaatt tccgggatca 300
aaagctgtta gcgaagcaag ttcaaacttt gggccttcct atgtatcggg cccaattatt 360
tttgtgttcg aaaaagtttc cactttcata gtcacagaag ataagaagga agaggaaccc 420
gcggctgctg atgacgtgca tgcaccagcc gccacgtcaa ctgaagaggt ggaggtgaag 480
gtgaaggaga aggagaagga gatagttatt gagtctggag aaccaaacaa ggaagaagca 540
cctgctactg ttgttgctga cgtggcacct gcaactaagg tggaagaagc accaaaggtg 600
gtttaa 606
<210>13
<211>750
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>13
atggctggcg gcgtagctat tggaagtttt agtgattcat tcagcgttgt ctctcttaag 60
tcctatcttg ccgaattcat ctccacactc atctttgtct tcgccggagt tggttccgcc 120
attgcttacg gcaagttgac aacaaatgct gcacttgatc cggctgggct tgtagctatt 180
gcagtttgcc atggatttgc tctattcgta gccgtttcga tttccgctaa catctccggt 240
ggtcatgtta accctgcggt cacctgtgga ttaaccttcg gcggacatat tacctttatc 300
actggctcct tctacatgct tgctcaactt accggcgccg ctgtagcttg cttcctcctc 360
aaattcgtca ccggaggatg tgctattcca acccatggag tgggagctgg tgtgagcata 420
ctagaaggac tcgtgatgga aataataatc acatttggtt tagtttatac tgtgttcgca 480
accgccgctg acccgaagaa gggttcattg ggcacaattg caccgattgc aattggtctc 540
attgttggag ctaatatttt ggctgccgga ccattctccg gtggatcaat gaacccagct 600
cgttcatttg gacctgcaat ggttagtggt aactttgagg gtttctggat ctactggatt 660
ggtccattag ttggtggtag tttggctggt cttatttaca caaatgtgtt catgacacaa 720
gaacatgctc ctttatccaa tgagttctaa 750
<210>14
<211>735
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>14
atggaggtcg attctagtgg gaatcctaat tggttatttg attatgagtt gatgacggat 60
attacttctg ctgcatctgt taccgtcgct gagtttcagt ctccggctac tattgatttc 120
agctggcctg ctcaaacgat ctatgcttct tctaatctca ttactgaaac agattacaca 180
tttgcggatt cagaagttag caaggaggca agctcacgaa agcggttaaa aagtgaatgt 240
tgcagctctc cgagatctaa ggcatgcaga gagaaattgc ggagggacag actgaatgag 300
aggttcctcg cattgagctc tgtccttgat cctggaaggc caccaaaaac tgagaaagtt 360
gcaattctaa gtgatgctca aaggatgctg attgagctgc gaactgaaac ccagaagctg 420
aaggagtcaa atgaggagct gcaagagaag ataaaagaac ttaaggcaga gaagaatgag 480
ctccgagatg aaaagcaaag gctaaaggaa gaaaaggata atttggagca gcaggttaaa 540
agcttagctt ctaaagcagg atttctctcc catccttctg ccatgggagc tgcatttact 600
gcacaaggac aagttgctgc aggcaacaaa ttgatgcctt tcattggtta tcccagygty 660
gcgatgtggc rattcatgca acctgctgtt gttgacacat ctcaagatca tgtgctccgt 720
cctccagttg cttaa 735
<210>15
<211>645
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>15
cccttcaaac ctaaaaaaaa gagaaaatca acaaaaaaaa tgggagcttg tgcaacgaag 60
ccaaaggatt tgaaaggaga tgcaccggaa accgcaccgg aaaatgttcc ggcgactgaa 120
atcgccacca aggatgcggc ggaagtagcc gtcgccgcca aggatgtggt ggttgtggcg 180
gaagtagaag tgaaaaagga aattgaggcc gatgctgctg ctgcagatga cgacgatgct 240
gaaaaacgcc gatctctatc taacttgttc aaagagaacg aagaatgtaa ggggtcagag 300
caagtaaacg aggaggcatc caagatcaca ccatcagaag ctaagccaga agaagttgag 360
aaggttgttg atgctcctgt aacttcagag atagaaaaag cactagaagt ggcctcaatt 420
actgctgagg ctcccaaggt ggagccttcc gaggagaaga agatagatga agtgaaatca 480
gaacctaaga ctcctgctga gaagaaagta gaggaagcaa aaccggcagt agagactcct 540
gcagagaaga aaatagaaga ggcaccagtt gctccaactc acgtggagac gaaagccgaa 600
gatgctccaa aggtaactgt agtagaagaa aagaagtcga gctag 645
<210>16
<211>630
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>16
cccttcaaac ctaaaaaaaa gagaaaatca acaaaaaaaa tgggagcttg tgcaacgaag 60
ccaaaggatt tgaaaggaga tgcaccggaa accgcaccgg aaaatgttcc ggcgactgaa 120
atcgccacca aggatgcggc ggaagtagcc gtcgccgcca aggatgtggt ggttgtggcg 180
gaagtagaag tgaaaaagga aattgaggcc gatgctgctg ctgcagatga cgacgatgct 240
gaaaaacgcc gatctctatc taacttgttc aaagagaacg aagaatgtaa ggggtcagag 300
caagtaaacg aggaggcatc caagatcaca ccatcagaag ctaagccaga agaagttgag 360
aaggttgttg atgctcctgt aacttcagag atagaaaaag cactagaagt ggcctcaatt 420
actgctgagg ctcccaaggt ggagccttcc gaggagaaga agatagatga agtgaaatca 480
gaacctaaga ctcctgctga gaagaaagta gaggaagcaa aaccggcagt agagactcct 540
gcagagaaga aaatagaaga ggcaccagtt gctccaactc acgtggagac gaaagccgaa 600
gattgcatgg tgaaaaacgc gtggaaatga 630
<210>17
<211>216
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>17
atggagataa catgtgcact tgtctgtgag atcttactag caatcttgct tcctcctctt 60
ggtgtttgcc ttcgcaatgg ttgctgcact gtggagttct tgatttgctt ggtattgacc 120
atattgggct atgttcctgg aattatctat gctctctatg caatcctctt tgttgaacgt 180
gaaccaagaa gggattatta tgacactctt gcttga 216
<210>18
<211>216
<212>DNA
<213>番茄(Lycopersicon esculentum)
<400>18
atggactcaa gatgtgcaat tctgtgtgaa gttttactag caatcttgct tcctcctctt 60
ggtgtttgcc ttcgcaatgg ttgctgcact gtggagttct tgatttgctt ggtattgacc 120
atattgggct atgttcctgg aattatctat gctctttatg caatcctctg tattcaaagt 180
gaaccacact accatcatta ccattctctt gcttga 216
<210>19
<211>877
<212>DNA
<213>花椰菜花叶病毒
<400>19
aagcttgcat gcctgcaggt ccccagatta gccttttcaa tttcagaaag aatgctaacc 60
cacagatggt tagagaggct tacgcagcag gtctcatcaa gacgatctac ccgagcaata 120
atctccagga aatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga 180
ctaactgcat caagaacaca gagaaagata tatttctcaa gatcagaagt actattccag 240
tatggacgat tcaaggcttg cttcacaaac caaggcaagt aatagagatt ggagtctcta 300
aaaaggtagt tcccactgaa tcaaaggcca tggagtcaaa gattcaaata gaggacctaa 360
cagaactcgc cgtaaagact ggcgaacagt tcatacagag tctcttacga ctcaatgaca 420
agaagaaaat cttcgtcaac atggtggagc acgacacact tgtctactcc aaaaatatca 480
aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg gtaatatccg 540
gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag atagtggaaa 600
aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc gttgaagatg 660
cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag 720
aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa 780
gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat 840
ttcatttgga gagaacacgg gggactctag aggatcc 877
<210>20
<211>2322
<212>DNA
<213>拟南芥(Arabidopsis thaliana)
<400>20
aagctttaag ctccaagccc acatctatgc acttcaacat atctttttct agatgagttg 60
gtaaaagtag aaaaagatat gatgatttta aatttgtttc tatttatatg tgttcatcga 120
aacttcattt tttttagttt taatagagag tttatatgac ttttaaaaat tgatttaaaa 180
ctgtgtcaaa aattaaaagg acaataaaaa atttgcatac aaccgaaaat acttatattt 240
agacaagaaa aaataatact tgtgatgctg attttatttt attatatatc atgaatcatg 300
atcatccaat tttccggata agccaaagtc aaaatgatgg gttcccccta atcttttatg 360
ctgagaaata gatgtatatt cttagatagt aatataaaat tgggttaaag aatgatgatt 420
cgattatagc ctcaactaga agatacgtgt agtgcaggtg tgtagttaac tggtggtagt 480
ggcagacaac cagattagga gttaaataaa gcctttagat ttgagagatt gaaatattcg 540
attggaacct ttctagattt ttacagccat ctaaaattag atgcagatca cctactacca 600
ttcaaaaatg aacaaaataa tttcatttac attttcctag cataagatat aataataaaa 660
tagtgctcat tttaattact ttttctaaat attttcgtta ttttaaattt tgcttgtcta 720
tactctacag ctcatttaat aacggaaaca aaaataattg cagggatacg gatgggtagc 780
tttcaaaact tacatcatct tctgtttctt gagatcaact atttttggag ctttgtctca 840
atcgtaccaa aggataatgg tcctacctcc ttttgcattc ttaactttat cttctctact 900
tatttctttt ttgggatttt tgggggtatt attttatctt ttgtagatat acacattgat 960
ttactacaaa cgtatactac tatccatctt caactcttcg gaatatgatt tcgaaaaaac 1020
tatgaagatt aacgggtatc ttaaacatgt taagatacac cggacaattt tcatttagaa 1080
gaattgatat gcaattaaca ataaatagtt gatgatcttt tagttttgaa gatgtgcgtt 1140
aagacttaag cgtgtggtaa caaggtggga ctcgggcaac gcaaagcctt gtagagtcca 1200
cttgctcaac ttgtctttct tttatctctt ttccaagtct caagattcaa tgaactccgt 1260
gtaacacaaa cacgcccata gatgagctca tttttggtat ttccaatatt gccactccat 1320
gataatatca tctagggatg gggttcattt attttgaaat ctcaacaaat ctcgtcgatt 1380
ctaacacaca tgattgattt gtttacttac ttgaaagttg gcaactatct gggattaaaa 1440
tttatctttt tctactgcta gctagaagca tctatatatg ttagcctaat acgtggaaga 1500
tgtcattgct aataatggct aaagatgtgt attaattttt cttctttttt ccttgaattt 1560
ttgttctttg acataaacta tgctgtcaaa atgtgtagaa tctttttaca taaatcattc 1620
cctgttacac actaaaaggt tcacaacgga cgattgtatt ggacttccag atcataaacc 1680
atgcaaaact gaaaaccaca agaataatta gttctaactt tagaacgttc gtacgtgttt 1740
catgttcaaa aagcgtcaat tataaaagtt gggaaattac ttttgagttt tgacatttct 1800
aaggacagtc aaatatgaca acattgggat gcaacttacc ttgtattaac ttattttgtt 1860
ataaaaccat atattacata ttttaaaggg ttgataaata atcaaatata ccaaaacata 1920
gcttttcaat atatttgtaa aacacgtttg gtctactagc taattatgag aacatttgtt 1980
caatgcatga ttatctagta tctactagtg gattatgaaa attagatatt ttcattgcat 2040
gattatcttc catatatagt gataacatca aaagaatcta caccaattat tgcatttttt 2100
cattatataa taagcactaa actgtaaaat tatattcagc cacccaaacc atgacaaatc 2160
accttaaagg cttaaacaca taacagccat tacgagtcac aggtaagggt ataatagtaa 2220
agaatcaatc tatataatat acgacccacc ctttctcatt ctttctggag agtaacatcg 2280
agacaaagaa gaaaaactaa aaaagagaac cccaaaggat cc 2322
<210>21
<211>31
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>21
gggaaggatc catgggtcgt atgcacagtc g 31
<210>22
<211>32
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>22
cgacggagct cctatgccac aagtgtgcta gc 32
<210>23
<211>35
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>23
gggaaggatc cggtggtatt gaagttatgg aagac 35
<210>24
<211>30
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>24
gctgcgagct cctaatgctt ccgtccactc 30
<210>25
<211>34
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>25
gggaaggatc catgaattct cagatttgca gatc 34
<210>26
<211>36
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>26
gaggagagct cttagttatc accatatagc cacttc 36
<210>27
<211>33
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>27
gggaatctag acaacatgga gaatatgcag agc 33
<210>28
<211>31
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>28
agaagcccgg gcacatagca cctacacacc g 31
<210>29
<211>33
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>29
gggaaggatc cgatgggata ctggaaagca aag 33
<210>30
<211>32
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>30
agccggagct cttaaaccac ctttggtgct tc 32
<210>31
<211>29
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>31
gggaaggatc caaatggctg gcggcgtag 29
<210>32
<211>34
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>32
agaaggagct ctagaactca ttggataaag gagc 34
<210>33
<211>32
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>33
gggaaggatc catggaggtc gattctagtg gg 32
<210>34
<211>32
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>34
agaagcccgg gttaagcaac tggaggacgg ag 32
<210>35
<211>22
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>35
ggtggctcct acaaatgcca tc 22
<210>36
<211>20
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>36
aagttgggta acgccagggt 20
<210>37
<211>22
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>37
tagtttggtc agatgggaaa cg 22
<210>38
<211>25
<212>DNA
<213>人工序列
<220>
<223>单链DNA寡核苷酸
<400>38
aaatattgga tcctttgggg ttctc 25
<210>39
<211>151
<212>PRT
<213>人参(Panax ginseng)
<400>39
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Thr Pro Pro Ser Trp Leu Lys Ile Thr Pro Gln Asp
20 25 30
Val Asp Asp Asn Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Ala Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>40
<211>151
<212>PRT
<213>大豆(Glycine max)
<400>40
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ser Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Thr Pro Pro Ser Trp Leu Lys Ile Ser Ser Gln Asp
20 25 30
Val Glu Glu Asn Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Ala Gln Val Asn
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ser Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>41
<211>151
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>41
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Ser Ser Pro Ser Trp Leu Lys Thr Thr Ser Gln Asp
20 25 30
Val Asp Glu Ser Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Pro Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>42
<211>151
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>42
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Ser Ser Pro Ser Trp Leu Lys Thr Thr Pro Gln Asp
20 25 30
Val Asp Glu Ser Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Pro Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>43
<211>151
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>43
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro His Lys Arg Ser Ser Pro Ser Trp Leu Lys Thr Thr Pro Gln Asp
20 25 30
Val Asp Glu Ser Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Pro Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>44
<211>151
<212>PRT
<213>豌豆(Pisum sativum)
<400>44
Met Gly Arg Leu His Ser Lys Gly Lys Gly Ile Ser Ser Ser Ala Leu
1 5 10 15
Pro Tyr Arg Arg Thr Ala Pro Ser Trp Leu Lys Ile Ser Ser Gln Asp
20 25 30
Val Asp Glu Thr Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Ala Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ser Ile Arg Lys His Leu Glu Arg Phe Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>45
<211>150
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>45
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Ser Ser Pro Ser Trp Leu Lys Thr Thr Ser Gln Asp
20 25 30
Val Asp Glu Ser Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Pro Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Asn Glu Ser Thr Thr
130 135 140
Ala Ser Thr Leu Val Ala
145 150
<210>46
<211>150
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>46
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ala Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Ser Ser Pro Ser Trp Leu Lys Thr Thr Pro Gln Asp
20 25 30
Val Asp Glu Ser Ile Cys Lys Phe Ala Lys Lys Gly Leu Thr Pro Ser
35 40 45
Gln Ile Gly Val Ile Leu Arg Asp Ser His Gly Ile Pro Gln Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr His Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Lys Thr Lys Lys Leu Pro Pro Val Trp Asn Glu Ser Thr Thr
130 135 140
Ala Ser Thr Leu Val Ala
145 150
<210>47
<211>151
<212>PRT
<213>稻(Oryza sativa)
<400>47
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ser Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Thr Pro Pro Ser Trp Leu Lys Thr Ala Ala Ser Asp
20 25 30
Val Glu Glu Met Ile Met Lys Ala Ala Lys Lys Gly Gln Met Pro Ser
35 40 45
Gln Ile Gly Val Val Leu Arg Asp Gln His Gly Ile Pro Leu Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr Phe Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Arg Thr Lys Lys Leu Pro Pro Thr Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>48
<211>151
<212>PRT
<213>玉蜀黍(Zea mays)
<400>48
Met Gly Ala Met His Ser Arg Gly Lys Gly Ile Ser Ser Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Thr Pro Pro Thr Trp Leu Lys Thr Ala Ala Ser Asp
20 25 30
Val Glu Glu Met Ile Thr Lys Ala Ala Lys Lys Gly Gln Met Pro Ser
35 40 45
Gln Ile Gly Val Leu Leu Arg Asp Gln His Gly Ile Pro Leu Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr Phe Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Arg Thr Lys Lys Leu Pro Pro Thr Trp Lys Tyr Glu Ser Thr
130 135 140
Thr Ala Ser Thr Leu Val Ala
145 150
<210>49
<211>156
<212>PRT
<213>稻(Oryza sativa)
<400>49
Met Gly Arg Met His Ser Arg Gly Lys Gly Ile Ser Ser Ser Ala Ile
1 5 10 15
Pro Tyr Lys Arg Thr Pro Pro Ser Trp Val Lys Thr Ala Ala Ala Asp
20 25 30
Val Glu Glu Met Ile Met Lys Ala Ala Lys Lys Gly Gln Met Pro Ser
35 40 45
Gln Ile Gly Val Val Leu Arg Asp Gln His Gly Ile Pro Leu Val Lys
50 55 60
Ser Val Thr Gly Ser Lys Ile Leu Arg Ile Leu Lys Ala His Gly Leu
65 70 75 80
Ala Pro Glu Ile Pro Glu Asp Leu Tyr Phe Leu Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Arg Thr Lys Lys Leu Pro Pro Thr Trp Lys Thr Thr Tyr Gln
130 135 140
Leu Ser Ala Val Cys Leu Asn Tyr Phe Gln Ala Pro
145 150 155
<210>50
<211>151
<212>PRT
<213>黑腹果蝇(Drosophila melanogaster)
<400>50
Met Gly Arg Met His Ala Pro Gly Lys Gly Ile Ser Gln Ser Ala Leu
1 5 10 15
Pro Tyr Arg Arg Thr Val Pro Ser Trp Leu Lys Leu Asn Ala Asp Asp
20 25 30
Val Lys Glu Gln Ile Lys Lys Ala Gly Gln Glu Gly Ser Asp Ser Leu
35 40 45
Gln Ile Gly Ile Ile Leu Arg Asp Ser His Gly Val Ala Gln Val Arg
50 55 60
Phe Val Asn Gly Asn Lys Ile Leu Arg Ile Met Lys Ser Val Gly Leu
65 70 75 80
Lys Pro Asp Ile Pro Glu Asp Leu Tyr His Met Ile Lys Lys Ala Val
85 90 95
Ala Ile Arg Lys His Leu Glu Arg Asn Arg Lys Asp Lys Asp Gly Lys
100 105 110
Phe Arg Leu Ile Leu Val Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Thr Lys Ser Val Leu Pro Pro Asn Trp Lys Tyr Glu Ser Ser
130 135 140
Thr Ala Ser Ala Leu Val Ala
145 150
<210>51
<211>155
<212>PRT
<213>未知
<220>
<223>环境序列
<400>51
Met Gly Arg Met His Thr Pro Gly Lys Gly Met Ser Gly Ser Ala Leu
1 5 10 15
Pro Tyr Lys Arg Ser Ala Pro Ser Trp Leu Lys Ile Thr Pro Thr Glu
20 25 30
Val Thr Glu Met Ile Val Lys Met Ala Lys Lys Gly Met Thr Pro Ser
35 40 45
Gln Ile Gly Val Met Leu Arg Asp Asn Gln Gly Ile Ala Gln Val Ser
50 55 60
Thr Val Thr Asn Ser Lys Ile Leu Arg Ile Leu Arg Gly Gln Gly Leu
65 70 75 80
Ala Pro Ser Leu Pro Glu Asp Leu Tyr Cys Leu Ile Lys Lys Ala Val
85 90 95
Ser Val Arg Lys His Leu Glu Arg Asn Arg Lys Asp Met Asp Ser Lys
100 105 110
Phe Arg Leu Ile Leu Ile Glu Ser Arg Ile His Arg Leu Ala Arg Tyr
115 120 125
Tyr Lys Leu Ala Lys Lys Leu Glu Pro Thr Phe Lys Tyr Asp Ser Ala
130 135 140
Thr Ala Ser Thr Leu Leu Thr Ala Ala Gly Lys
145 150 155
<210>52
<211>230
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>52
Met Ala Ser Ser Asn Ser Glu Lys Ile Asn Glu Asn Leu Tyr Ala Val
1 5 10 15
Leu Gly Leu Lys Lys Glu Cys Ser Lys Thr Glu Leu Arg Ser Ala Tyr
20 25 30
Lys Lys Leu Ala Leu Arg Trp His Pro Asp Arg Cys Ser Ser Met Glu
35 40 45
Phe Val Glu Glu Ala Lys Lys Lys Phe Gln Ala Ile Gln Glu Ala Tyr
50 55 60
Ser Val Leu Ser Asp Ser Asn Lys Arg Phe Leu Tyr Asp Val Gly Ala
65 70 75 80
Tyr Asn Thr Asp Asp Asp Asp Asp Gln Asn Gly Met Gly Asp Phe Leu
85 90 95
Asn Glu Met Ala Thr Met Met Asn Gln Ser Lys Pro Ser Asp Asn Asn
100 105 110
Thr Gly Asp Ser Phe Glu Gln Leu Gln Asp Leu Phe Asn Glu Met Phe
115 120 125
Gln Gly Asp Ala Ala Ala Phe Pro Ser Ser Ser Ser Cys Ser Thr Ser
130 135 140
Asn Phe Thr Ser Ser Arg Ser Phe Val Phe Asp Thr Asn Ser Gln Arg
145 150 155 160
Ser Ser Ser Phe Ala Thr Ser Ser Met Gly Met Asn Asn Asp Pro Phe
165 170 175
Gly Tyr Asp Pro Arg Ala His Ser Phe Ser Leu Gly Val Asp His Gln
180 185 190
Gln Glu Phe Lys Lys Gly Lys Asn Asn Gly Gly Arg Arg Asn Arg Arg
195 200 205
Lys Asn Asn Val Pro Ser Ala Gly His Glu Thr Ser Ser Ser Asn Asn
210 215 220
Tyr Gly Val Pro Thr Ser
225 230
<210>53
<211>242
<212>PRT
<213>小家鼠(Mus musculus)
<400>53
Met Val Asp Tyr Tyr Glu Val Leu Gly Val Gln Arg His Ala Ser Pro
1 5 10 15
Glu Asp Ile Lys Lys Ala Tyr Arg Lys Gln Ala Leu Lys Trp His Pro
20 25 30
Asp Lys Asn Pro Glu Asn Lys Glu Glu Ala Glu Arg Lys Phe Lys Gln
35 40 45
Val Ala Glu Ala Tyr Glu Val Leu Ser Asp Ala Lys Lys Arg Asp Ile
50 55 60
Tyr Asp Lys Tyr Gly Lys Glu Gly Leu Asn Gly Gly Gly Gly Gly Gly
65 70 75 80
Gly Ile His Phe Asp Ser Pro Phe Glu Phe Gly Phe Thr Phe Arg Asn
85 90 95
Pro Asp Asp Val Phe Arg Glu Phe Phe Gly Gly Arg Asp Pro Phe Ser
100 105 110
Phe Asp Phe Phe Glu Asp Pro Phe Asp Asp Phe Phe Gly Asn Arg Arg
115 120 125
Gly Pro Arg Gly Asn Arg Ser Arg Gly Ala Ala Pro Phe Phe Ser Thr
130 135 140
Phe Ser Gly Phe Pro Ser Phe Gly Ser Gly Phe Pro Ala Phe Asp Thr
145 150 155 160
Gly Phe Thr Pro Phe Gly Ser Leu Gly His Gly Gly Leu Thr Ser Phe
165 170 175
Ser Ser Thr Ser Phe Gly Gly Ser Gly Met Gly Asn Phe Lys Ser Ile
180 185 190
Ser Thr Ser Thr Lys Ile Val Asn Gly Lys Lys Ile Thr Thr Lys Arg
195 200 205
Ile Val Glu Asn Gly Gln Glu Arg Val Glu Val Glu Glu Asp Gly Gln
210 215 220
Leu Lys Pro Leu Thr Ile Asn Gly Lys Glu His Leu Leu Arg Leu Asp
225 230 235 240
Asn Lys
<210>54
<211>242
<212>PRT
<213>褐鼠(Rattus norvegicus)
<400>54
Met Val Asp Tyr Tyr Glu Val Leu Gly Val Gln Arg His Ala Ser Pro
1 5 10 15
Glu Asp Ile Lys Lys Ala Tyr Arg Lys Gln Ala Leu Lys Trp His Pro
20 25 30
Asp Lys Asn Pro Glu Asn Lys Glu Glu Ala Glu Arg Lys Phe Lys Gln
35 40 45
Val Ala Glu Ala Tyr Glu Val Leu Ser Asp Ala Lys Lys Arg Asp Ile
50 55 60
Tyr Asp Lys Tyr Gly Lys Glu Gly Leu Asn Gly Gly Gly Gly Gly Gly
65 70 75 80
Gly Ser His Phe Asp Ser Pro Phe Glu Phe Gly Phe Thr Phe Arg Asn
85 90 95
Pro Asp Asp Val Phe Arg Glu Phe Phe Gly Gly Arg Asp Pro Phe Ser
100 105 110
Phe Asp Phe Phe Glu Asp Pro Phe Asp Asp Phe Phe Gly Asn Arg Arg
115 120 125
Gly Pro Arg Gly Ser Arg Ser Arg Gly Ala Gly Ser Phe Phe Ser Ala
130 135 140
Phe Ser Gly Phe Pro Ser Phe Gly Ser Gly Phe Pro Ala Phe Asp Thr
145 150 155 160
Gly Phe Thr Pro Phe Gly Ser Leu Gly His Gly Gly Leu Thr Ser Phe
165 170 175
Ser Ser Ala Ser Phe Gly Gly Ser Gly Met Gly Asn Phe Lys Ser Ile
180 185 190
Ser Thr Ser Thr Lys Ile Val Asn Gly Lys Lys Ile Thr Thr Lys ATg
195 200 205
Ile Val Glu Asn Gly Gln Glu Arg Val Glu Val Glu Glu Asp Gly Gln
210 215 220
Leu Lys Ser Leu Thr Ile Asn Gly Lys Glu His Leu Leu Arg Leu Asp
225 230 235 240
Asn Lys
<210>55
<211>158
<212>PRT
<213>白云杉(Picea glauca)
<400>55
Met Glu Thr Thr Phe Tyr Ser Ile Leu Gly Val Asn Lys Asp Ser Ser
1 5 10 15
Ser Ala Glu Ile Arg Ser Ala Tyr Arg Lys Leu Ala Met Lys Trp His
20 25 30
Pro Asp Lys Trp Ser Thr Asp Pro Ser Ser Ser Glu Thr Ala Lys Leu
35 40 45
Arg Phe Gln Gln Ile Gln Glu Ala Tyr Ser Val Leu Ser Asp Asp Thr
50 55 60
Lys Arg Ala Leu Tyr Asp Ala Gly Met Tyr Glu Pro Ser Glu Asp Met
65 70 75 80
Asp Ala Phe Cys Asp Phe Leu Asp Glu Leu Ser Ser Leu Ile Ala Thr
85 90 95
Val Lys Val Gln Ser Asn Lys Asp Asp Glu Leu Leu Gln Leu Gln Glu
100 105 110
Met Phe Thr Lys Met Leu Glu Glu Asp Trp Phe Ser Thr Asp Asn Phe
115 120 125
Glu Ala Phe Lys Glu Ile Ser Ser Gln His Ser Asp Asp Lys Pro Glu
130 135 140
Asn Gly Gln Asp His Glu Pro Tyr Gly Ser Val Asp Asp Leu
145 150 155
<210>56
<211>156
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>56
Met Ala Cys Glu Gly Gly Gly Ser Asn Val Arg Ser Ser Tyr Tyr Thr
1 5 10 15
Ile Leu Gly Ile Arg Lys Asp Ala Ser Val Ser Asp Ile Arg Thr Ala
20 25 30
Tyr Arg Lys Leu Ala Met Lys Trp His Pro Asp Arg Tyr Ala Arg Asn
35 40 45
Pro Gly Val Ala Gly Glu Ala Lys Arg Arg Phe Gln Gln Ile Gln Glu
50 55 60
Ala Tyr Ser Val Leu Asn Asp Glu Asn Lys Arg Ser Met Tyr Asp Val
65 70 75 80
Gly Leu Tyr Asp Pro His Glu Asp Asp Asp Asp Asp Phe Cys Asp Phe
85 90 95
Met Gln Glu Met Ile Ser Met Met Asn Asn Val Lys Asp Ala Gly Glu
100 105 110
Ser Leu Glu Asp Leu Gln Arg Met Phe Thr Asp Met Val Gly Gly Asp
115 120 125
Gly Val Ser Tyr Asp Cys Asn Asn Asn Pro Lys Gly Asn Lys Arg Pro
130 135 140
Arg Val Asn Ile Ser Arg Ser Ser Ala Ala Met Arg
145 150 155
<210>57
<211>156
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>57
Met Ala Cys Glu Gly Gly Gly Ser Asn Val Arg Ser Ser Tyr Tyr Thr
1 5 10 15
Val Leu Gly Ile Arg Lys Asp Ala Ser Val Ser Asp Ile Arg Thr Ala
20 25 30
Tyr Arg Lys Leu Ala Met Lys Trp His Pro Asp Arg Tyr Ala Arg Asn
35 40 45
Pro Gly Val Ala Gly Glu Ala Lys Arg Arg Phe Gln Gln Ile Gln Glu
50 55 60
Ala Tyr Ser Val Leu Asn Asp Glu Asn Lys Arg Ser Met Tyr Asp Val
65 70 75 80
Gly Leu Tyr Asp Pro His Glu Asp Asp Asp Asp Asp Phe Cys Asp Phe
85 90 95
Met Gln Glu Met Ile Ser Met Met Asn Asn Val Lys Asp Glu Gly Glu
100 105 110
Ser Leu Glu Asp Leu Gln Arg Met Phe Thr Asp Met Val Gly Gly Asp
115 120 125
Gly Val Ser Tyr Asp Cys Asn Asn Asn Pro Lys Gly Ser Lys Arg Pro
130 135 140
Arg Val Asn Val Ser Arg Ser Ser Ala Ala Met Arg
145 150 155
<210>58
<211>207
<212>PRT
<213>胡萝卜(Daucus carota)
<400>58
Met Ile Asp Gln Glu Glu Ser Asn Phe Asn Phe Asn Phe Asn Gln Pro
1 5 10 15
Gln Gln Pro Gln Gln Gln Gln Phe His Gly Lys Ser Val Lys Lys Asn
20 25 30
Lys Asn Lys Asn Asn Asn Asn Asn Ser Glu Ser Gly Asn Lys Asn Gly
35 40 45
Gly Glu Asn Lys Asn Gly Val Glu Lys Arg Phe Lys Thr Leu Pro Pro
50 55 60
Ala Glu Ser Leu Pro Arg Asn Glu Thr Val Gly Gly Tyr Ile Phe Val
65 70 75 80
Cys Asn Asn Asp Thr Met Gln Glu Asn Leu Lys Arg Gln Leu Phe Gly
85 90 95
Leu Pro Pro Arg Tyr Arg Asp Ser Val Arg Ala Ile Thr Pro Gly Leu
100 105 110
Pro Leu Phe Leu Tyr Asn Tyr Ser Thr His Gln Leu His Gly Val Phe
115 120 125
Glu Ala Ala Ser Phe Gly Gly Thr Asn Ile Asp Pro Thr Ala Trp Glu
130 135 140
Asp Lys Lys Asn Gln Gly G1u Ser Arg Phe Pro Ala Gln Val Arg Val
145 150 155 160
Met Thr Arg Lys Ile Cys Glu Pro Leu Glu Glu Asp Ser Phe Arg Pro
165 170 175
Ile Leu His His Tyr Asp Gly Pro Lys Phe Arg Leu Glu Leu Asn Ile
180 185 190
Pro Glu Ala Ile Ser Leu Leu Asp Ile Phe Glu Glu Thr Lys Ala
195 200 205
<210>59
<211>213
<212>PRT
<213>豌豆(Pisum sativum)
<400>59
Met Asn Lys Asn Ser Leu Arg Asn Gly Val Tyr Asn Met Asn Ala Val
1 5 10 15
Tyr Gln Lys Ser Asn Ala Asn Phe Val Gly Asn Met Asn Ser Asn Lys
20 25 30
Tyr Ser Gly Asn Val Gln Leu Asn Lys Asp Pro His Ser Asn Asn Asn
35 40 45
Asn Asn Asn Asn Glu Asn Asn Thr Asn Ala Thr Asp Lys Arg Phe Lys
50 55 60
Thr Leu Pro Ala Ala Glu Thr Leu Pro Arg Asn Glu Val Leu Gly Gly
65 70 75 80
Tyr Ile Phe Val Cys Asn Asn Asp Thr Met Gln Glu Asp Leu Lys Arg
85 90 95
Gln Leu Phe Gly Leu Pro Pro Arg Tyr Arg Asp Ser Val Arg Ala Ile
100 105 110
Thr Pro Gly Leu Pro Leu Phe Leu Tyr Asn Tyr Thr Thr His Gln Leu
115 120 125
His Gly Ile Phe Glu Ala Thr Cys Phe Gly Gly Ser Asn Ile Asp Pro
130 135 140
Thr Ala Trp Glu Asp Lys Lys Cys Lys Gly Glu Ser Arg Phe Pro Ala
145 150 155 160
Gln Val Arg Ile Arg Val Arg Lys Ile Cys Lys Ala Leu Glu Glu Asp
165 170 175
Ser Phe Arg Pro Val Leu His His Tyr Asp Gly Pro Lys Phe Arg Leu
180 185 190
Glu Leu Ser Val Pro Glu Thr Leu Asp Leu Met Asp Leu Cys Glu Gln
195 200 205
Ala Gly Ser Ala Ala
210
<210>60
<211>305
<212>PRT
<213>葡萄柚(Citrus X paradisi)
<400>60
Met Asp Asn Met His Ser Phe Trp Gln Leu Gly Asp Glu Leu Arg Gly
1 5 10 15
Gln Ser Arg Thr Ser Glu Asp Gln Ser Trp Leu Arg Ala Ala Ser Arg
20 25 30
Leu Ala Glu Gln Thr Arg Phe Lys Gly Glu Arg Met Asn Asn Leu Asp
35 40 45
Leu Ser Lys Gly Met Thr Glu Ile Arg Pro Arg Asp Lys Ile Met Tyr
50 55 60
His Glu Asp Asn Asn Phe Glu Ser Phe Asn Phe Asn Phe Asn Met Met
65 70 75 80
Asn Leu Asp Asn Lys Val Val Glu Asn Val Thr Lys Ser Ser Leu Arg
85 90 95
Asn Gly Ile Tyr Asn Met Asn Ala Val Tyr Gln Lys Asn Ser Gly His
100 105 110
Asn Met Gly Asn Leu Met Val Asn Lys Tyr Gly Gly Asn Asn Leu Ser
115 120 125
Val Lys Glu Ala Glu Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asp
130 135 140
Ser Asn Ala Asn Ser Ala Leu Asp Lys Arg Phe Lys Thr Leu Pro Ala
145 150 155 160
Thr Glu Thr Leu Pro Arg Asn Glu Val Leu Gly Gly Tyr Ile Phe Val
165 170 175
Cys Asn Asn Asp Thr Met Gln Glu Asp Leu Lys Arg Gln Leu Phe Gly
180 185 190
Leu Pro Pro Arg Tyr Arg Asp Ser Val Arg Ala Ile Thr Pro Gly Leu
195 200 205
Pro Leu Phe Leu Tyr Asn Tyr Thr Thr His Gln Leu His Gly Ile Phe
210 215 220
Glu Ala Thr Gly Phe Gly Gly Ser Asn Ile Asp Pro Thr Ala Trp Glu
225 230 235 240
Asp Lys Lys Cys Lys Gly Glu Ser Arg Phe Pro Ala Gln Val Arg Ile
245 250 255
Arg Val Arg Lys Leu Cys Lys Ala Leu Glu Glu Asp Ala Phe Arg Pro
260 265 270
Val Leu His His Tyr Asp Gly Pro Lys Phe Arg Leu Glu Leu Ser Val
275 280 285
Pro Glu Thr Leu Asp Leu Met Asp Leu Cys Glu Gln Ala Gly Ser Ala
290 295 300
Ala
305
<210>61
<211>296
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>61
Met Asp Ser Phe Trp Gln Leu Gly Asp Glu Leu Arg Gly Gln Thr Arg
1 5 10 15
Ala Ser Glu Asp His Lys Trp Ser Thr Val Ala Thr Lys Leu Ala Glu
20 25 30
Gln Thr Arg Met Lys Gly Glu Arg Met Asn Asn Leu Asp Leu Ser Lys
35 40 45
Gly Tyr Thr Glu Phe Arg Pro Ser Glu Lys Phe Ser Phe Gln Glu Asn
50 55 60
Asn Leu Asn Phe Asn Met Leu Asn Leu Asp Gly Lys Phe Gly Glu Ser
65 70 75 80
Ile Met Gly Lys Thr Ser Met Gln Ser Asn Val Tyr Asn Met Asn Thr
85 90 95
Val Phe Gln Lys Asn Asp Phe Lys Ser Gly Gly Asn Met Lys Val Asn
100 105 110
Lys Tyr Asn Gly Asn Ile Val Ala Asn Lys Glu Met Ser Asn Asn Lys
115 120 125
His Asn Asn Asn Cys Asn Asp Asn Gly Asn Met Asn Leu Ala Val Asp
130 135 140
Lys Arg Phe Lys Thr Leu Pro Ala Ser Glu Thr Leu Pro Arg Asn Glu
145 150 155 160
Val Leu Gly Gly Tyr Ile Phe Val Cys Asn Asn Asp Thr Met Gln Glu
165 170 175
Asp Met Lys Arg His Leu Phe Gly Leu Pro Pro Arg Tyr Arg Asp Ser
180 185 190
Val Arg Ala Ile Thr Pro Gly Leu Pro Leu Phe Leu Tyr Asn Tyr Thr
195 200 205
Thr His Gln Leu His Gly Ile Phe Glu Ala Thr Thr Phe Gly Gly Thr
210 215 220
Asn Ile Asp Ala Thr Ala Trp Glu Asp Lys Lys Cys Lys Gly Glu Ser
225 230 235 240
Arg Phe Pro Ala Gln Val Arg Ile Arg Val Arg Lys Ile Cys Lys Ala
245 250 255
Leu Glu Glu Asp Ser Phe Arg Pro Val Leu His His Tyr Asp Gly Pro
260 265 270
Lys Phe Arg Leu Glu Leu Ser Val Pro Glu Thr Leu Asp Leu Leu Asp
275 280 285
Leu Cys Glu Gln Ala Gly Ser Ala
290 295
<210>62
<211>296
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>62
Met Asp Ser Phe Trp Gln Leu Gly Asp Glu Leu Arg Gly Gln Thr Arg
1 5 10 15
Ala Ser Glu Asp His Lys Trp Ser Thr Val Ala Thr Lys Leu Ala Glu
20 25 30
Gln Thr Arg Met Lys Gly Glu Arg Met Asn Asn Leu Asp Leu Ser Lys
35 40 45
Gly Tyr Thr Glu Phe Arg Pro Ser Glu Lys Phe Ser Phe Gln Glu Asn
50 55 60
Asn Leu Asn Phe Asn Met Leu Asn Leu Asp Gly Lys Phe Gly Glu Ser
65 70 75 80
Ile Met Gly Lys Thr Ser Met Gln Ser Asn Val Tyr Asn Met Asn Thr
85 90 95
Val Phe Gln Lys Asn Asp Phe Lys Ser Gly Gly Asn Met Lys Val Asn
100 105 110
Lys Tyr Asn Gly Asn Val Val Ala Asn Lys Glu Met Ser Asn Asn Lys
115 120 125
His Asn Asn Asn Cys Asn Asp Asn Gly Asn Met Asn Leu Ala Val Asp
130 135 140
Lys Arg Phe Lys Thr Leu Pro Ala Ser Glu Thr Leu Pro Arg Asn Glu
145 150 155 160
Val Leu Gly Gly Tyr Ile Phe Val Cys Asn Asn Asp Thr Met Gln Glu
165 170 175
Asp Met Lys Arg His Leu Phe Gly Leu Pro Pro ATg Tyr Arg Asp Ser
180 185 190
Val Arg Ala Ile Thr Pro Gly Leu Pro Leu Phe Leu Tyr Asn Tyr Thr
195 200 205
Thr His Gln Leu His Gly Ile Phe Glu Ala Thr Thr Phe Gly Gly Thr
210 215 220
Asn Ile Asp Ala Thr Ala Trp Glu Asp Lys Lys Cys Lys Gly Glu Ser
225 230 235 240
Arg Phe Pro Ala Gln Val Arg Ile Arg Val Arg Lys Ile Cys Lys Ala
245 250 255
Leu Glu Glu Asp Ser Phe Arg Pro Val Leu His His Tyr Asp Gly Pro
260 265 270
Lys Phe Arg Leu Glu Leu Ser Val Pro Glu Thr Leu Asp Leu Leu Asp
275 280 285
Leu Cys Glu Gln Ala Gly Ser Ala
290 295
<210>63
<211>742
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>63
Met Ala Glu Ala Met Glu Thr Glu Met Asp Phe Ser Asp Gly Glu Gln
1 5 10 15
Thr Asn Gly Asn Ser His Val Thr Ala Ser Gln Tyr Phe Ala Pro Pro
20 25 30
Gly Tyr Asn Arg Ser Leu Val Ala Ala Tyr Gly Asn Gly Asn Thr Thr
35 40 45
Ile Gly Leu Glu Lys Gly Ile Glu Arg Arg Leu Asp His His Glu Gln
50 55 60
Leu Pro Gly Tyr Ile Phe Met Cys Asn Gly Arg Thr Lys Thr Asp Cys
65 70 75 80
Tyr Arg Tyr Arg Val Phe Gly Ile Pro Arg Gly Gly Lys Asp Val Val
85 90 95
Glu Ser Ile Lys Pro Gly Met Lys Leu Phe Leu Tyr Asp Phe Glu Lys
100 105 110
Arg Leu Leu Tyr Gly Val Tyr Glu Ala Thr Val Gly Gly Arg Leu Asp
115 120 125
Ile Glu Pro Glu Ala Phe Glu Gly Lys Tyr Pro Ala Gln Val Gly Phe
130 135 140
Arg Ile Val Met Asn Cys Leu Pro Leu Thr Glu Asn Thr Phe Lys Ser
145 150 155 160
Ala Ile Tyr Glu Asn Tyr Lys Gly Ser Lys Phe Lys Gln Glu Leu Ser
165 170 175
Pro His Gln Val Met Ser Leu Leu Ser Leu Phe Arg Ser Phe Thr Ser
180 185 190
Pro Glu Leu Asp Leu Leu Pro His Arg Leu Ala Ser Arg Ala Ser Ala
195 200 205
Pro Arg Thr Leu Ser Phe Glu Glu Arg Phe Ile Ala Ala Thr His Leu
210 215 220
Arg Asn Ala Ser Ser Val Leu Asp Pro Leu Ser Ala Arg His Val Glu
225 230 235 240
Pro Arg Leu Gly Ser Val Met Ala His Gln Pro Val Pro Arg Thr Ser
245 250 255
Leu Leu Gln His Ser Tyr Phe Arg Gln Asp Asp Tyr Thr Thr Pro Pro
260 265 270
Arg Glu Ser Leu Ser Asn Leu Asn Gln Pro Tyr Tyr Pro Thr Glu Ala
275 280 285
Arg Gln Leu Arg Leu Leu Gly Asp Pro Ser Arg Ser Asp Ser Pro Arg
290 295 300
Ser Glu Pro Pro Arg Ser Ser Ile Gln Asp Pro Gln Leu Lys Tyr Leu
305 310 315 320
Thr Ile Leu Ser Asn Ile Arg Arg Tyr Gly Ser Ala Ser Asp Arg Leu
325 330 335
Ala Ser Glu Asn Glu Tyr His Pro Ala Thr Pro Ser Glu Lys Asp Gln
340 345 350
Phe Ala Val Pro Tyr Ser Asp Asn Lys Asn Tyr Pro Ser Thr Leu Ser
355 360 365
Gly Ser Glu His Pro Ser Ala Ser Ala Ala Asn Gly Ser Val Tyr Arg
370 375 380
Ser Glu Phe Tyr Asn Ser Ala Ser Gln Lys Glu Gly Glu Ala Ser Gln
385 390 395 400
Gln His Glu Ile Pro Ala Gly Thr Tyr His His Pro Glu Ala Ser Thr
405 410 415
Val Ser Asn Thr Thr Lys Ser Met Gln Pro Asp Met Gln Ala Val Ser
420 425 430
Val Ala Gln Ser His Thr Glu Thr Ala Gly Tyr Pro Thr Pro Ala His
435 440 445
Gly Glu Ala Ser Gln Pro Pro Ala Gly Ala Ile Gly Tyr Thr His Gln
450 455 460
Pro Gln Ser Val Ala Gly Asn Tyr Ser Thr His Ser Gln Pro Gly Asn
465 470 475 480
Val Glu Glu Ser Thr Gln Ser Tyr Ala Gly Thr Asp Ser Tyr Ser Gln
485 490 495
Gln Gln Tyr Tyr Ala Ala Met Gly Pro Thr Thr Gln Leu His Ala Gly
500 505 510
Gly Tyr Ile Gln Lys Pro His Glu Ile Gly Tyr Ser Gln Gln Pro His
515 520 525
Asp Ala Ala Thr Gly Tyr Ser Gln Gln Pro His Asp Ala Ala Thr Gly
530 535 540
Tyr Ser Gln Gln Pro His Asp Ala Ala Thr Gly Tyr Ser Gln Gln Pro
545 550 555 560
His Ala Ala Ser Thr Gly Tyr Ser Gln Gln Thr Tyr Ala Ala Ala Thr
565 570 575
Gly Tyr Thr Gln Gln Pro His Ala Ala Ala Ala Gly Tyr Thr Gln Gln
580 585 590
Pro His Ala Ala Ala Thr Gly Tyr Ser Gln Gln Pro His Ala Ala Ala
595 600 605
Thr Ala His Ala Gln Gln Pro Tyr Ala Ala Ala Thr Ala His Ala Gln
610 615 620
Gln Leu His Ala Val Ala Thr Gly Tyr Ala Leu Gln Leu His Ala Ala
625 630 635 640
Ala Thr Gly Tyr Ala Gln Gln Pro His Ala Ala Ala Thr Gly Tyr Ala
645 650 655
Leu Gln Pro His Ala Gln Ala Val Glu Tyr Thr Met Gln Pro His Ala
660 665 670
Gln Ala Val Gly Tyr Met Pro Gln Tyr His Ala His Ala Val Val Tyr
675 680 685
Ser Gln Gln Gly Val Thr Gln Gly Ser Val Pro Arg Ala Pro Gly Thr
690 695 700
Thr Asp Cys Asn Ala Ala Asn Gln Ala Tyr Ser Ala Thr Gly Asp Trp
705 710 715 720
Asn Ala Val His Gln Ser Tyr Tyr Pro Gln Thr Ala Asp Ala Thr Thr
725 730 735
Thr Tyr Tyr Gln Thr Ser
740
<210>64
<211>195
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>64
Met Ala Gly Val Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Leu Ala
1 5 10 15
Ser Leu Arg Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Thr Pro Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Ile Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Val Ile Thr Gly Val Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Ala Ala Cys Phe Leu Leu Lys Tyr Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Ser Val Ala Ala Gly Leu Gly Ser Ile Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Leu Ala
165 170 175
Ile Gly Leu Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser
195
<210>65
<211>248
<212>PRT
<213>陆地棉(Gossypium hirsutum)
<400>65
Met Ala Gly Ile Ala Phe Gly Arg Phe Asp Asp Ser Phe Ser Leu Gly
1 5 10 15
Thr Val Lys Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Val Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Thr Asp
35 40 45
Ala Ala Leu Asp Pro Asp Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Ile Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Leu Gly Gly Gln Ile
85 90 95
Thr Ile Leu Thr Gly Ile Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Ile Val Ala Cys Phe Leu Leu Lys Ala Val Thr Gly Gly Leu Thr Val
115 120 125
Pro Ile His Gly Leu Gly Ala Gly Val Gly Ala Ile Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser
195 200 205
Gly Asp Phe Asn Gly Ile Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ser Gly Leu Ile Tyr Gly Asn Val Phe Met Asn Ser Asp
225 230 235 240
His Ala Pro Leu Ser Asn Asp Phe
245
<210>66
<211>250
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>66
Met Ala Gly Val Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Leu Ala
1 5 10 15
Ser Leu Arg Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Thr Pro Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Ile Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Val Ile Thr Gly Val Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Ala Ala Cys Phe Leu Leu Lys Tyr Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Ser Val Ala Ala Gly Leu Gly Ser Ile Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Leu Ala
165 170 175
Ile Gly Leu Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala
195 200 205
Gly Asp Phe Ser Gly His Trp Val Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Glu Leu Ala Gly Leu Ile Tyr Gly Asn Val Phe Met Gly Ser Ser
225 230 235 240
Glu His Val Pro Leu Ala Ser Ala Asp Phe
245 250
<210>67
<211>250
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>67
Met Ala Gly Val Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Leu Ala
1 5 10 15
Ser Leu Arg Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Thr Pro Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Ile Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Val Ile Thr Gly Val Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Ala Ala Cys Phe Leu Leu Lys Tyr Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Ser Val Ala Ala Gly Leu Gly Ser Ile Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Leu Ala
165 170 175
Ile Gly Leu Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala
195 200 205
Gly Asp Phe Ser Gly His Trp Val Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Leu Ile Tyr Gly Asn Val Phe Met Gly Ser Ser
225 230 235 240
Glu His Val Pro Leu Ala Ser Ala Asp Phe
245 250
<210>68
<211>250
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>68
Met Val Lys Ile Glu Ile Gly Ser Val Gly Asp Ser Phe Ser Val Ala
1 5 10 15
Ser Leu Lys Ala Tyr Leu Ser Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Leu Ala Phe Ala Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Val Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Ile Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Val Ile Thr Gly Phe Phe Tyr Trp Ile Ala Gln Cys Leu Gly Ser
100 105 110
Ile Val Ala Cys Leu Leu Leu Val Phe Val Thr Asn Gly Glu Ser Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Leu Gly Ala Ile Glu Gly Val Val
130 135 140
Met Glu Ile Val Val Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ser
195 200 205
Gly Asp Phe Ser Gln Ile Trp Ile Tyr Trp Val Gly Pro Leu Val Gly
210 215 220
Gly Ala Leu Ala Gly Leu Ile Tyr Gly Asp Val Phe Ile Gly Ser Tyr
225 230 235 240
Ala Pro Ala Pro Thr Thr Glu Ser Tyr Pro
245 250
<210>69
<211>248
<212>PRT
<213>光烟草(Nicotiana glauca)
<400>69
Met Pro Gly Ile Ala Phe Gly Arg Ile Asp Asp Ser Phe Ser Val Gly
1 5 10 15
Ser Leu Lys Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ala Asn
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Val Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Val Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Leu Gly Gly Gln Ile
85 90 95
Thr Leu Leu Thr Gly Leu Phe Tyr Ile Ile Ala Gln Leu Leu Gly Ser
100 105 110
Ile Val Ala Cys Leu Leu Leu Lys Val Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Asn Val Ala Ala Gly Val Gly Ala Leu Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser
195 200 205
Gly Asp Phe Thr Asn Asn Trp Ile Tyr Trp Ala Gly Pro Leu Val Gly
210 215 220
Gly Gly Leu Ala Gly Leu Thr Tyr Ser Asn Val Phe Met Gln His Glu
225 230 235 240
His Ala Pro Leu Ser Ser Asp Phe
245
<210>70
<211>248
<212>PRT
<213>萝卜(Raphanus sativus)
<400>70
Met Ala Gly Val Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Leu Ala
1 5 10 15
Ser Leu Arg Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Thr Pro Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Ile Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Leu Ile Thr Gly Val Phe Tyr Trp Val Ala Gln Leu Leu Gly Ser
100 105 110
Thr Ala Ala Cys Phe Leu Leu Lys Tyr Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Ser Val Ala Ala Gly Val Gly Ala Ile Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ser Leu Val Tyr Thr Val Tyr Pro Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Leu Ala
165 170 175
Ile Gly Leu Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala
195 200 205
Gly Asp Phe Ser Gly His Trp Val Tyr Trp Val Gly Pro Leu Ile Gly
2l0 215 220
Gly Gly Leu Ala Gly Val Thr Tyr Gly Asn Val Phe Met Thr Ser Glu
225 230 235 240
His Val Pro Leu Ala Ser Glu Phe
245
<210>71
<211>248
<212>PRT
<213>向日葵(Helianthus annuus)
<400>71
Met Pro Gly Ile Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Leu Ser
1 5 10 15
Ser Ile Lys Ser Tyr Val Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Phe Ala Lys Leu Thr Ala Asp
35 40 45
Ala Asp Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Leu Ala Leu Phe Val Ala Val Ser Ile Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Ile Leu Thr Gly Ile Phe Tyr Trp Ile Ala Gln Cys Ile Gly Ser
100 105 110
Ile Ala Ala Cys Tyr Leu Leu Ser Phe Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Ala Val Ala Ala Gly Val Gly Ala Ile Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Asn Val Tyr Ala Thr
145 150 155 160
Ala Val Asp Pro Lys Lys Gly Asp Leu Gly Thr Ile Ala Pro Leu Ala
165 170 175
Ile Gly Leu Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala
195 200 205
Gly Asp Phe Ser Gly His Trp Ile Tyr Trp Val Gly Pro Leu Val Gly
210 215 220
Gly Gly Leu Ala Gly Ala Ile Tyr Ser Asn Val Phe Ile Ser Asn Glu
225 230 235 240
His Ala Pro Leu Ser Ser Glu Phe
245
<210>72
<211>248
<212>PRT
<213>稻(Oryza sativa)
<400>72
Met Val Lys Leu Ala Phe Gly Ser Leu Gly Asp Ser Phe Ser Ala Thr
1 5 10 15
Ser Val Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Gly Gln Leu Thr Asn Gly
35 40 45
Gly Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Leu Ala Leu Phe Val Gly Val Ser Val Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly His Ile
85 90 95
Thr Ile Leu Thr Gly Leu Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ala
100 105 110
Ser Ile Ala Cys Leu Leu Leu Lys Phe Val Thr His Gly Lys Ala Ile
115 120 125
Pro Thr His Gly Val Ala Gly Ile Ser Glu Leu Glu Gly Val Val Met
130 135 140
Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr Ala
145 150 155 160
Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala Ile
165 170 175
Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser Gly
180 185 190
Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala Gly
195 200 205
Asn Phe Ala Gly Asn Trp Val Tyr Trp Val Gly Pro Leu Ile Gly Gly
210 215 220
Gly Leu Ala Gly Leu Val Tyr Gly Asp Val Phe Ile Gly Ser Tyr Gln
225 230 235 240
Pro Val Ala Asp Gln Asp Tyr Ala
245
<210>73
<211>248
<212>PRT
<213>玉蜀黍(Zea mays)
<400>73
Met Val Lys Leu Ala Phe Gly Ser Phe Arg Asp Ser Leu Ser Ala Ala
1 5 10 15
Ser Leu Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ser Gln Leu Thr Lys Gly
35 40 45
Gly Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly His Ile
85 90 95
Thr Ile Leu Thr Gly Ile Leu Tyr Trp Val Ala Gln Leu Leu Gly Ala
100 105 110
Ser Val Ala Cys Phe Leu Leu Gln Tyr Val Thr His Gly Gln Ala Ile
115 120 125
Pro Thr His Gly Val Ser Gly Ile Ser Glu Ile Glu Gly Val Val Met
130 135 140
Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr Ala
145 150 155 160
Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Met Ala Ile
165 170 175
Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser Gly
180 185 190
Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala Gly
195 200 205
Asn Phe Ala Gly Asn Trp Val Tyr Trp Val Gly Pro Leu Val Gly Gly
210 215 220
Gly Leu Ala Gly Leu Val Tyr Gly Asp Val Phe Ile Ala Ser Tyr Gln
225 230 235 240
Pro Val Gly Gln Gln Glu Tyr Pro
245
<210>74
<211>249
<212>PRT
<213>大麦(Hordeum vulgare)
<400>74
Met Val Lys Leu Ala Phe Gly Ser Phe Gly Asp Ser Phe Ser Ala Thr
1 5 10 15
Ser Ile Arg Ser Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ser Tyr Gly Gln Leu Thr Gln Gly
35 40 45
Gly Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ala Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly His Val
85 90 95
Thr Ile Leu Thr Gly Leu Phe Tyr Trp Val Ala Gln Leu Leu Gly Ala
100 105 110
Ser Val Ala Cys Leu Leu Leu Gln Phe Val Thr His Ala Gln Ala Met
115 120 125
Pro Thr His Ala Val Ser Gly Ile Ser Glu Val Glu Gly Val Val Met
130 135 140
Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr Ala
145 150 155 160
Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Met Ala Ile
165 170 175
Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser Gly
180 185 190
Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala Gly
195 200 205
Asn Phe Ser Gly His Trp Val Tyr Trp Val Gly Pro Leu Ile Gly Gly
210 215 220
Gly Leu Ala Gly Leu Val Tyr Gly Asp Val Phe Ile Ala Ser Tyr Gln
225 230 235 240
Pro Val Gly His Gln Gln Glu Tyr Pro
245
<210>75
<211>228
<212>PRT
<213>金鱼草(Antirrhinum majus)
<400>75
Phe Gly Ser Ile Gly Asp Ser Phe Ser Val Ala Ser Ile Lys Ala Tyr
1 5 10 15
Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val Phe Ala Gly Val Gly
20 25 30
Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ser Asp Ala Ala Leu Asp Pro
35 40 45
Ala Gly Leu Val Ala Val Ala Val Ala His Ala Phe Ala Leu Phe Val
50 55 60
Gly Val Ser Met Ala Ala Asn Val Ser Gly Gly His Leu Asn Pro Ala
65 70 75 80
Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile Thr Ile Leu Thr Gly
85 90 95
Leu Phe Tyr Trp Ile Ala Gln Cys Leu Gly Ser Thr Val Ala Cys Leu
100 105 110
Leu Leu Lys Phe Val Thr Asn Gly Leu Ser Val Pro Thr His Gly Val
115 120 125
Ala Ala Gly Met Asp Ala Ile Gln Gly Val Val Met Glu Ile Ile Ile
130 135 140
Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr Ala Ala Asp Pro Lys
145 150 155 160
Lys Gly Ser Leu Gly Val Ile Ala Pro Ile Ala Ile Gly Phe Ile Val
165 170 175
Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser Gly Gly Ser Met Asn
180 185 190
Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser Gly Asp Phe Ser Gln
195 200 205
Asn Trp Ile Tyr Trp Ala Gly Pro Leu Ile Gly Gly Ala Leu Ala Gly
210 215 220
Phe Ile Tyr Gly
225
<210>76
<211>248
<212>PRT
<213>玉蜀黍(Zea mays)
<400>76
Met Val Lys Leu Ala Phe Gly Ser Phe Arg Asp Ser Leu Ser Ala Ala
1 5 10 15
Ser Leu Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ser Gln Leu Thr Lys Gly
35 40 45
Gly Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly His Ile
85 90 95
Thr Ile Leu Thr Gly Ile Leu Tyr Trp Val Ala Gln Leu Leu Gly Ala
100 105 110
Ser Val Ala Cys Phe Leu Leu Gln Tyr Val Thr His Gly Gln Ala Ile
115 120 125
Pro Thr His Gly Val Ser Gly Ile Ser Glu Ile Glu Gly Val Val Met
130 135 140
Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr Ala
145 150 155 160
Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Met Ala Ile
165 170 175
Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser Gly
180 185 190
Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ala Gly
195 200 205
Asn Phe Ala Gly Asn Trp Val Tyr Trp Val Gly Pro Leu Val Gly Gly
210 215 220
Gly Leu Ala Gly Pro Val Tyr Gly Asp Val Phe Ile Ala Ser Tyr Gln
225 230 235 240
Pro Val Gly Gln Gln Glu Tyr Pro
245
<210>77
<211>248
<212>PRT
<213>向日葵(Helianthus annuus)
<400>77
Met Pro Gly Ile Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Ser Ser
1 5 10 15
Ser Ile Lys Ala Tyr Ile Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ala Asp
35 40 45
Ala Ala Leu Asp Pro Pro Gly Leu Val Ala Val Ala Val Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Cys Ile Ala Ala Asn Ile Cys Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Phe Leu Thr Gly Leu Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Val Ala Cys Phe Leu Leu Ser Phe Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Glu Gly Val Gly Thr Ile Gln Gly Val Val
130 135 140
Phe Glu Ile Ile Ile Thr Phe Ala Met Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Cys Asp Pro Lys Lys Gly Ala Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser
195 200 205
Phe Asp Phe Ser Gly His Trp Ile Tyr Trp Val Gly Pro Leu Val Gly
210 215 220
Gly Gly Leu Ala Gly Leu Ile Tyr Pro Asn Val Phe Ile Ser Asn Glu
225 230 235 240
His Ile Pro Leu Thr Asn Glu Tyr
245
<210>78
<211>248
<212>PRT
<213>向日葵(Helianthus annuus)
<400>78
Met Pro Gly Ile Ala Phe Gly Ser Phe Asp Asp Ser Phe Ser Ser Ser
1 5 10 15
Ser Ile Lys Ala Tyr Ile Ala Glu Phe Ile Ser Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Ala Lys Leu Thr Ala Asp
35 40 45
Ala Ala Leu Asp Pro Ser Gly Leu Val Ala Ile Ala Val Cys His Gly
50 55 60
Leu Ala Leu Phe Val Ala Val Ser Ile Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Thr Phe Gly Leu Ala Val Gly Gly Gln Ile
85 90 95
Thr Ile Leu Thr Gly Leu Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Val Ala Ala Cys Phe Leu Leu Ser Phe Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Val Gly Val Leu Gln Gly Val Val
130 135 140
Phe Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Cys Asp Pro Lys Lys Gly A1a Leu Gly Thr Ile Ser Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Thr Val Ala Ala
195 200 205
Phe Asp Phe Ser Gly His Trp Phe Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Ala Ile Tyr Pro Asn Val Phe Ile Ser Asn Glu
225 230 235 240
His Ile Pro Leu Thr Asn Asp Tyr
245
<210>79
<211>248
<212>PRT
<213>油桐(Vernicia fordii)
<400>79
Met Ala Arg Ile Ala Phe Gly Arg Phe Asp Asp Ser Phe Ser Leu Gly
1 5 10 15
Ser Phe Lys Ala Tyr Leu Ala Glu Phe Ile Ser Thr Leu Leu Tyr Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ala Asn
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Cys His Gly
50 55 60
Phe Ala Leu Phe Val Ala Val Ala Val Gly Ala Asn Ile Ser Gly Gly
65 70 75 80
His Val Asn Pro Ala Val Ala Phe Gly Leu Ala Leu Gly Gly Gln Ile
85 90 95
Thr Ile Leu Thr Gly Ile Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Ile Val Ala Cys Phe Leu Leu Lys Leu Val Thr Gly Gly Leu Ala Ile
115 120 125
Pro Thr His Ser Val Ala Gly Arg Val Gly Ala Ile Glu Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe IIe Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Tyr Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ser
195 200 205
Gly Asp Phe His Asp Asn Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Leu Ile Tyr Gly Asn Leu Tyr Ile Ser Gly Asp
225 230 235 240
His Thr Pro Leu Ser Asn Asp Phe
245
<210>80
<211>248
<212>PRT
<213>胡萝卜(Daucus carota)
<400>80
Met Val Lys Leu Ala Ile Gly Ser Val Gly Asp Ser Phe Ser Ala Val
1 5 10 15
Ser Leu Lys Ser Tyr Leu Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Gly His Leu Thr Ala Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Ile Gly Gly Asn Ile
85 90 95
Thr Ile Ile Thr Gly Leu Phe Tyr Trp Ile Ala Gln Cys Leu Gly Ser
100 105 110
Thr Val Ala Cys Phe Leu Leu Lys Phe Val Thr Ala Gly Lys Ala Ile
115 120 125
Pro Thr His Gly Val Gly Ala Gly Leu Gly Ala Ala Glu Gly Val Val
130 135 140
Phe Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser
195 200 205
Phe Asp Phe Ser Gly His Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Leu Val Tyr Gly Asp Val Phe Ile Gly Ser Tyr
225 230 235 240
Ala Pro Ile Ala Glu Asp Tyr Ala
245
<210>81
<211>250
<212>PRT
<213>金鱼草(Antirrhinum majus)
<400>81
Met Val Lys Ile Ala Phe Gly Ser Ile Gly Asp Ser Phe Ser Val Ala
1 5 10 15
Ser Ile Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Val Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Val Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Ile Leu Thr Gly Leu Phe Tyr Trp Ile Ala Gln Cys Leu Gly Ser
100 105 110
Thr Val Ala Cys Leu Leu Leu Lys Phe Val Thr Asn Gly Leu Ser Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Met Asp Ala Ile Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Val Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Ala Ser
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Ala Gly Pro Leu Ile Gly
210 215 220
Gly Ala Leu Ala Gly Phe Ile Tyr Gly Asp Val Phe Ile Thr Ala His
225 230 235 240
Ala Pro Leu Pro Thr Ser Glu Asp Tyr Ala
245 250
<210>82
<211>250
<212>PRT
<213>烟草(Nicotiana tabacum)
<400>82
Met Val Arg Ile Ala Phe Gly Ser Ile Gly Asp Ser Phe Ser Val Gly
1 5 10 15
Ser Leu Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ala Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Val Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Ile Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Ile Leu Thr Gly Phe Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Val Ala Cys Leu Leu Leu Lys Tyr Val Thr Asn Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Leu Asn Gly Leu Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ala
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Ala Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Phe Ile Tyr Gly Asp Val Phe Ile Gly Cys His
225 230 235 240
Thr Pro Leu Pro Thr Ser Glu Asp Tyr Ala
245 250
<210>83
<211>250
<212>PRT
<213>向日葵(Helianthus annuus)
<400>83
Met Val Lys Leu Ala Ile Gly Ser Ile Gly Asp Ser Leu Ser Ala Gly
1 5 10 15
Ser Ile Lys Ser Tyr Leu Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Gly Lys Leu Thr Thr Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Ile Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Phe Gly Leu Ala Ile Gly Gly Asn Ile
85 90 95
Thr Ile Ile Thr Gly Leu Phe Tyr Trp Ile Ala Gln Cys Leu Gly Ser
100 105 110
Ile Val Ala Cys Phe Leu Leu Gln Phe Val Thr Gly Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Asp Gly Met Asn Gly Val Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Val Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Met Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ser
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Phe Ile Phe Gly Asp Val Phe Ile Gly Ser Tyr
225 230 235 240
Glu Thr Leu Pro Asp Ser Gly Asp Tyr Ala
245 250
<210>84
<211>250
<212>PRT
<213>马铃薯(Solanum tuberosum)
<400>84
Met Val Arg Ile Ala Phe Gly Ser Ile Gly Asp Ser Leu Ser Val Gly
1 5 10 15
Ser Leu Lys Ala Tyr Leu Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ser Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Thr Leu Thr Gly Leu Phe Tyr Trp Val Ala Gln Leu Leu Gly Ser
100 105 110
Thr Val Ala Cys Leu Leu Leu Lys Tyr Val Thr Asn Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Met Asn Gly Ala Glu Gly Val Val
130 135 140
Met Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ala
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Phe Ile Tyr Gly Asp Val Phe Ile Gly Ser His
225 230 235 240
Thr Pro Leu Pro Thr Ser Glu Asp Tyr Ala
245 250
<210>85
<211>250
<212>PRT
<213>烟草(Nicotiana tabacum)
<400>85
Met Val Leu Ile Ala Phe Gly Ser Ile Gly Asp Ser Phe Ser Val Gly
1 5 10 15
Ser Leu Lys Ala Tyr Val Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Tyr Asn Lys Leu Thr Ala Asp
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Val Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Ile Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Ile Leu Thr Gly Phe Phe Tyr Trp Ile Ala Gln Leu Leu Gly Ser
100 105 110
Thr Val Ala Cys Leu Leu Leu Lys Tyr Val Thr Asn Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Leu Asn Gly Phe Gln Gly Val Val
130 135 140
Met Glu Ile Ile Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ala
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Ala Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Phe Ile Tyr Gly Asp Val Phe Ile Gly Cys His
225 230 235 240
Thr Pro Leu Pro Thr Ser Glu Asp Tyr Ala
245 250
<210>86
<211>250
<212>PRT
<213>番茄(Lycopersicon esculentum)
<400>86
Met Val Arg Ile Ala Phe Gly Ser Ile Gly Asp Ser Leu Ser Val Gly
1 5 10 15
Ser Leu Lys Ala Tyr Leu Ala Glu Phe Ile Ala Thr Leu Leu Phe Val
20 25 30
Phe Ala Gly Val Gly Ser Ala Ile Ala Phe Asn Lys Leu Thr Ser Gly
35 40 45
Ala Ala Leu Asp Pro Ala Gly Leu Val Ala Ile Ala Val Ala His Ala
50 55 60
Phe Ala Leu Phe Val Gly Val Ser Met Ala Ala Asn Ile Ser Gly Gly
65 70 75 80
His Leu Asn Pro Ala Val Thr Leu Gly Leu Ala Val Gly Gly Asn Ile
85 90 95
Thr Ile Leu Thr Gly Leu Phe Tyr Trp Val Ala Gln Leu Leu Gly Ser
100 105 110
Thr Val Ala Cys Leu Leu Leu Lys Tyr Val Thr Asn Gly Leu Ala Val
115 120 125
Pro Thr His Gly Val Ala Ala Gly Met Ser Gly Ala Glu Gly Val Val
130 135 140
Met Glu Ile Val Ile Thr Phe Ala Leu Val Tyr Thr Val Tyr Ala Thr
145 150 155 160
Ala Ala Asp Pro Lys Lys Gly Ser Leu Gly Thr Ile Ala Pro Ile Ala
165 170 175
Ile Gly Phe Ile Val Gly Ala Asn Ile Leu Ala Ala Gly Pro Phe Ser
180 185 190
Gly Gly Ser Met Asn Pro Ala Arg Ser Phe Gly Pro Ala Val Val Ala
195 200 205
Gly Asp Phe Ser Gln Asn Trp Ile Tyr Trp Val Gly Pro Leu Ile Gly
210 215 220
Gly Gly Leu Ala Gly Phe Ile Tyr Gly Asp Val Phe Ile Gly Cys His
225 230 235 240
Thr Pro Leu Pro Thr Ser Glu Asp Tyr Ala
245 250
<210>87
<21l>234
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>87
Met Val Ser Pro Glu Asn Ala Asn Trp Ile Cys Asp Leu Ile Asp Ala
1 5 10 15
Asp Tyr Gly Ser Phe Thr Ile Gln Gly Pro Gly Phe Ser Trp Pro Val
20 25 30
Gln Gln Pro Ile Gly Val Ser Ser Asn Ser Ser Ala Gly Val Asp Gly
35 40 45
Ser Ala Gly Asn Ser Glu Ala Ser Lys Glu Pro Gly Ser Lys Lys Arg
50 55 60
Gly Arg Cys Glu Ser Ser Ser Ala Thr Ser Ser Lys Ala Cys Arg Glu
65 70 75 80
Lys Gln Arg Arg Asp Arg Leu Asn Asp Lys Phe Met Glu Leu Gly Ala
85 90 95
Ile Leu Glu Pro Gly Asn Pro Pro Lys Thr Asp Lys Ala Ala Ile Leu
100 105 110
Val Asp Ala Val Arg Met Val Thr Gln Leu Arg Gly Glu Ala Gln Lys
115 120 125
Leu Lys Asp Ser Asn Ser Ser Leu Gln Asp Lys Ile Lys Glu Leu Lys
130 135 140
Thr Glu Lys Asn Glu Leu Arg Asp Glu Lys Gln Arg Leu Lys Thr Glu
145 150 155 160
Lys Glu Lys Leu Glu Gln Gln Leu Lys Ala Met Asn Ala Pro Gln Pro
165 170 175
Ser Phe Phe Pro Ala Pro Pro Met Met Pro Thr Ala Phe Ala Ser Ala
180 185 190
Gln Gly Gln Ala Pro Gly Asn Lys Met Val Pro Ile Ile Ser Tyr Pro
195 200 205
Gly Val Ala Met Trp Gln Phe Met Pro Pro Ala Ser Val Asp Thr Ser
210 215 220
Gln Asp His Val Leu Arg Pro Pro Val Ala
225 230
<210>88
<211>234
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>88
Met Val Ser Pro Glu Asn Ala Asn Trp Ile Cys Asp Leu Ile Asp Ala
1 5 10 15
Asp Tyr Gly Ser Phe Thr Ile Gln Gly Pro Gly Phe Ser Trp Pro Val
20 25 30
Gln Gln Pro Ile Gly Val Ser Ser Asn Ser Ser Ala Gly Val Asp GIy
35 40 45
Ser Ala Gly Asn Ser Glu Ala Ser Lys Glu Pro Gly Ser Lys Lys Arg
50 55 60
Gly Arg Cys Glu Ser Ser Ser Ala Thr Ser Ser Lys Ala Cys Arg Glu
65 70 75 80
Lys Gln Arg Arg Asp Arg Leu Asn Asp Lys Phe Met Glu Leu Gly Ala
85 90 95
Ile Leu Glu Pro Gly Asn Pro Pro Lys Thr Asp Lys Ala Ala Ile Leu
100 105 110
Val Asp Ala Val Arg Met Val Thr Gln Leu Arg Gly Glu Ala Gln Lys
115 120 125
Leu Lys Asp Ser Asn Ser Ser Leu Gln Asp Lys Ile Lys Glu Leu Lys
130 135 140
Thr Glu Lys Asn Glu Leu Arg Asp Glu Lys Gln Arg Leu Lys Thr Glu
145 150 155 160
Lys Glu Lys Leu Glu Gln Gln Leu Lys Ala Ile Asn Ala Pro Gln Pro
165 170 175
Ser Phe Phe Pro Ala Pro Pro Met Met Pro Thr Ala Phe Ala Ser Ala
180 185 190
Gln Gly Gln Ala Pro Gly Asn Lys Met Val Pro Ile Ile Ser Tyr Pro
195 200 205
Gly Val Ala Met Trp Gln Phe Met Pro Pro Ala Ser Val Asp Thr Ser
210 215 220
Gln Asp His Val Leu Arg Pro Pro Val Ala
225 230
<210>89
<211>216
<212>PRT
<213>稻(Oryza sativa)
<400>89
Cys Gly Ala Phe Pro Trp Asp Ala Ser Pro Ser Cys Ser Asn Pro Ser
1 5 10 15
Val Glu Val Ser Ser Tyr Val Asn Thr Thr Ser Tyr Val Leu Lys Glu
20 25 30
Pro Gly Ser Asn Lys Arg Val Arg Ser Gly Ser Cys Gly Arg Pro Thr
35 40 45
Ser Lys Ala Ser Arg Glu Lys Ile Arg Arg Asp Lys Met Asn Asp Arg
50 55 60
Phe Leu Glu Leu Gly Thr Thr Leu Glu Pro Gly Lys Pro Val Lys Ser
65 70 75 80
Asp Lys Ala Ala Ile Leu Ser Asp Ala Thr Arg Met Val Ile Gln Leu
85 90 95
Arg Ala Glu Ala Lys Gln Leu Lys Asp Thr Asn Glu Ser Leu Glu Asp
100 105 110
Lys Ile Lys Glu Leu Lys Ala Glu Lys Asp Glu Leu Arg Asp Glu Lys
115 120 125
Gln Lys Leu Lys Val Glu Lys Glu Thr Leu Glu Gln Gln Val Lys Ile
130 135 140
Leu Thr Ala Thr Pro Ala Tyr Met Pro His Pro Thr Leu Met Pro Ala
145 150 155 160
Pro Tyr Pro Gln Ala Pro Leu Ala Pro Phe His His Ala Gln Gly Gln
165 170 175
Ala Ala Gly Gln Lys Leu Met Met Pro Phe Val Gly Tyr Pro Gly Tyr
180 185 190
Pro Met Trp Gln Phe Met Pro Pro Ser Glu Val Asp Thr Ser Lys Asp
195 200 205
Ser Glu Ala Cys Pro Pro Val Ala
210 215
<210>90
<211>226
<212>PRT
<213>拟南芥(Arabidopsis thaliana)
<400>90
Met Val Ser Pro Glu Asn Thr Asn Trp Leu Ser Asp Tyr Pro Leu Ile
1 5 10 15
Glu Gly Ala Phe Ser Asp Gln Asn Pro Thr Phe Pro Trp Gln Ile Asp
20 25 30
Gly Ser Ala Thr Val Ser Val Glu Val Asp Gly Phe Leu Cys Asp Ala
35 40 45
Asp Val Ile Lys Glu Pro Ser Ser Arg Lys Arg Ile Lys Thr Glu Ser
50 55 60
Cys Thr Gly Ser Asn Ser Lys Ala Cys Arg Glu Lys Gln Arg Arg Asp
65 70 75 80
Arg Leu Asn Asp Lys Phe Thr Glu Leu Ser Ser Val Leu Glu Pro Gly
85 90 95
Arg Thr Pro Lys Thr Asp Lys Val Ala Ile Ile Asn Asp Ala Ile Arg
100 105 110
Met Val Asn Gln Ala Arg Asp Glu Ala Gln Lys Leu Lys Asp Leu Asn
115 120 125
Ser Ser Leu Gln Glu Lys Ile Lys Glu Leu Lys Asp Glu Lys Asn Glu
130 135 140
Leu Arg Asp Glu Lys Gln Lys Leu Lys Val Glu Lys Glu Arg Ile Asp
145 150 155 160
Gln Gln Leu Lys Ala Ile Lys Thr Gln Pro Gln Pro Gln Pro Cys Phe
165 170 175
Leu Pro Asn Pro Gln Thr Leu Ser Gln Ala Gln Ala Pro Gly Ser Lys
180 185 190
Leu Val Pro Phe Thr Thr Tyr Pro Gly Phe Ala Met Trp Gln Phe Met
195 200 205
Pro Pro Ala Ala Val Asp Thr Ser Gln Asp His Val Leu Arg Pro Pro
210 215 220
Val Ala
225
<210>91
<211>256
<212>PRT
<213>稻(Oryza sativa)
<400>91
Met Ala Ser Pro Glu Gly Ser Thr Trp Val Phe Asp Cys Pro Leu Met
1 5 10 15
Asp Asp Leu Ala Ala Ala Ala Gly Phe Asp Ala Ala Pro Ala Gly Gly
20 25 30
Phe Tyr Trp Thr Thr Pro Ala Pro Pro Gln Ala Ala Leu Gln Pro Pro
35 40 45
Pro Pro Gln Gln Gln Pro Val Ala Pro Ala Thr Ala Ala Pro Asn Ala
50 55 60
Cys Ala Glu Ile Asn Gly Ser Val Asp Cys Glu His Gly Lys Glu Gln
65 70 75 80
Pro Thr Asn Lys Arg Pro Arg Ser Glu Ser Gly Thr Arg Pro Ser Ser
85 90 95
Lys Ala Cys Arg Glu Lys Val Arg Arg Asp Lys Leu Asn Glu Arg Phe
100 105 110
Leu Glu Leu Gly Ala Val Leu Glu Pro Gly Lys Thr Pro Lys Met Asp
115 120 125
Lys Ser Ser Ile Leu Asn Asp Ala Ile Arg Val Met Ala Glu Leu Arg
130 135 140
Ser Glu Ala Gln Lys Leu Lys Glu Ser Asn Glu Ser Leu Gln Glu Lys
145 150 155 160
Ile Lys Glu Leu Lys Ala Glu Lys Asn Glu Leu Arg Asp Glu Lys Gln
165 170 175
Lys Leu Lys Ala Glu Lys Glu Ser Leu Glu Gln Gln Ile Lys Phe Leu
180 185 190
Asn Ala Arg Pro Ser Phe Val Pro His Pro Pro Val Ile Pro Ala Ser
195 200 205
Ala Phe Thr Ala Pro Gln Gly Gln Ala Ala Gly Gln Lys Leu Met Met
210 215 220
Pro Val Ile Gly Tyr Pro Gly Phe Pro Met Trp Gln Phe Met Pro Pro
225 230 235 240
Ser Asp Val Asp Thr Thr Asp Asp Thr Lys Ser Cys Pro Pro Val Ala
245 250 255
<210>92
<21l>253
<212>PRT
<213>稻(Oryza sativa)
<400>92
Met Ser Gly Thr Pro Ala Asp Gly Gly Gly Gly Gly Gly Gly Gly Gly
1 5 10 15
Gly Gly Ser Gly Asp Asp Trp Phe Leu Asp Cys Gly Ile Leu Glu Asp
20 25 30
Leu Pro Ala Ala Ala Cys Gly Ala Phe Pro Trp Asp Ala Ser Pro Ser
35 40 45
Cys Ser Asn Pro Ser Val Glu Val Ser Ser Tyr Val Asn Thr Thr Ser
50 55 60
Tyr Val Leu Lys Glu Pro Gly Ser Asn Lys Arg Val Arg Ser Gly Ser
65 70 75 80
Cys Gly Arg Pro Thr Ser Lys Ala Ser Arg Glu Lys Ile Arg Arg Asp
85 90 95
Lys Met Asn Asp Arg Phe Leu Glu Leu Gly Thr Thr Leu Glu Pro Gly
100 105 110
Lys Pro Val Lys Ser Asp Lys Ala Ala Ile Leu Ser Asp Ala Thr Arg
115 120 125
Met Val Ile Gln Leu Arg Ala Glu Ala Lys Gln Leu Lys Asp Thr Asn
130 135 140
Glu Ser Leu Glu Asp Lys Ile Lys Glu Leu Lys Ala Glu Lys Asp Glu
145 150 155 160
Leu Arg Asp Glu Lys Gln Lys Leu Lys Val Glu Lys Glu Thr Leu Glu
165 170 175
Gln Gln Val Lys Ile Leu Thr Ala Thr Pro Ala Tyr Met Pro His Pro
180 185 190
Thr Leu Met Pro Ala Pro Tyr Pro Gln Ala Pro Leu Ala Pro Phe His
195 200 205
His Ala Gln Gly Gln Ala Ala Gly Gln Lys Leu Met Met Pro Phe Val
210 215 220
Gly Tyr Pro Gly Tyr Pro Met Trp Gln Phe Met Pro Pro Ser Glu Val
225 230 235 240
Asp Thr Ser Lys Asp Ser Glu Ala Cys Pro Pro Val Ala
245 250
Claims (36)
1.一种提高植物对非生物胁迫的耐受性的方法,该方法包括在植物中表达外源多核苷酸,该外源多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有90%同源,因而能提高植物对非生物胁迫的耐受性。
2.权利要求1的方法,其中所述表达通过下述步骤来实现:
(a)用所述外源多核苷酸转化所述植物的细胞;
(b)从所述细胞产生成熟植株;
(c)在适合所述成熟植株中表达所述外源多核苷酸的条件下,栽培所述成熟植株。
3.权利要求2的方法,其中所述转化是通过将核酸构建体导入所述植物细胞中来实现,该核酸构建体包括所述外源多核苷酸和至少一个能够在所述植物细胞中指导所述外源多核苷酸转录的启动子。
4.权利要求3的方法,其中所述至少一个启动子是组成型启动子。
5.权利要求3的方法,其中所述组成型启动子是花椰菜花叶病毒(CaMV)35S启动子。
6.权利要求3的方法,其中所述组成型启动子是At6669启动子。
7.权利要求3的方法,其中所述至少一个启动子是诱导型启动子。
8.权利要求7的方法,其中所述诱导型启动子是非生物胁迫诱导型启动子。
9.权利要求1的方法,其中所述表达是通过用包括所述外源多核苷酸的病毒感染所述植物来实现。
10.权利要求9的方法,其中所述病毒是无毒病毒。
11.权利要求1的方法,其中所述非生物胁迫选自高盐、失水、低温、高温、重金属毒性、无氧生活、营养缺乏、营养过剩、大气污染和紫外辐射。
12.权利要求1的方法,其中所述植物是双子叶植物。
13.权利要求1的方法,其中所述植物是单子叶植物。
14.一种提高植物生物量的方法,该方法包括在植物中表达外源多核苷酸,该外源多核苷酸与选自SEQ ID NO:1-18的多核苷酸至少有90%同源,因而能提高植物的生物量。
15.权利要求1的方法,其中所述表达通过下述步骤来实现:
(a)用所述外源多核苷酸转化所述植物的细胞;
(b)从所述细胞产生成熟植株;
(c)在适合所述成熟植株中表达所述外源多核苷酸的条件下,栽培所述成熟植株。
16.权利要求15的方法,其中所述转化是通过将核酸构建体导入所述植物细胞中来实现,该核酸构建体包括所述外源多核苷酸和至少一个能够在所述植物细胞中指导所述外源多核苷酸转录的启动子。
17.权利要求16的方法,其中所述至少一个启动子是组成型启动子。
18.权利要求17的方法,其中所述组成型启动子是CaMV 35S启动子。
19.权利要求17的方法,其中所述组成型启动子是At6669启动子。
20.权利要求16的方法,其中所述至少一个启动子是诱导型启动了。
21.权利要求14的方法,其中所述表达是通过用包括所述外源多核苷酸的病毒感染所述植物来实现。
22.权利要求21的方法,其中所述病毒是无毒病毒。
23.权利要求14的方法,其中所述植物是双子叶植物。
24.权利要求14的方法,其中所述植物是单子叶植物。
25.一种核酸构建体,该核酸构建体包括与选自SEQ ID NO:1-18的核苷酸序列至少有90%同源的多核苷酸和能够在宿主细胞中指导所述多核苷酸转录的启动子。
26.权利要求25的方法,其中所述启动子是组成型启动子。
27.权利要求26的方法,其中所述组成型启动子是CaMV 35S启动子。
28.权利要求26的方法,其中所述组成型启动子是At6669启动子。
29.权利要求25的方法,其中所述启动子是诱导型启动子。
30.权利要求29的方法,其中所述诱导型启动子是非生物胁迫诱导型启动子。
31.权利要求25的方法,其中所述宿主细胞是植物细胞。
32.权利要求31的方法,其中所述植物细胞成为双子叶植物细胞的一部分。
33.权利要求31的方法,其中所述植物细胞成为单子叶植物细胞的一部分。
34.一种分离的多肽,所述多肽包含与选自SEQ ID NO:1-18的多核苷酸编码的氨基酸序列至少有90%同源的氨基酸序列。
35.一种植物细胞,所述植物细胞包含与选自SEQ ID NO:1-18的多核苷酸至少有90%同源的外源多核苷酸。
36.权利要求35的植物细胞,其中所述植物细胞成为植物的一部分。
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Also Published As
Publication number | Publication date |
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ZA200509383B (en) | 2007-02-28 |
RU2005140106A (ru) | 2006-07-27 |
BRPI0411182A (pt) | 2006-07-18 |
EP1625199B1 (en) | 2015-03-11 |
CA2978152A1 (en) | 2004-12-02 |
RU2350653C2 (ru) | 2009-03-27 |
WO2004104162A3 (en) | 2005-12-15 |
EP1625199A2 (en) | 2006-02-15 |
RU2008143335A (ru) | 2010-05-10 |
CN101942449A (zh) | 2011-01-12 |
CA2526440C (en) | 2017-10-24 |
EP2365087A2 (en) | 2011-09-14 |
MXPA05012565A (es) | 2006-05-25 |
CN101942449B (zh) | 2014-07-02 |
CA2978152C (en) | 2021-01-26 |
CA2526440A1 (en) | 2004-12-02 |
WO2004104162A2 (en) | 2004-12-02 |
BR122016024209B1 (pt) | 2019-08-20 |
EP2365087A3 (en) | 2011-11-30 |
BRPI0411182B1 (pt) | 2019-08-20 |
CN100591770C (zh) | 2010-02-24 |
EP1625199A4 (en) | 2007-11-28 |
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