CN102337275B - 用于dmo的有效靶向的叶绿体转运肽及其用途 - Google Patents
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Abstract
本发明提供了用于有效加工和定位转基因植物中的麦草畏单加氧酶(DMO)的某些叶绿体转运肽的鉴定和用途。本发明也提供了产生耐受麦草畏的植物的方法、控制杂草生长的方法和产生食物、饲料和其它产品的方法,以及当出土前或出土后施用麦草畏时提供麦草畏耐受性的种子。
Description
本申请是申请日为2007年6月6日、申请号为200780052290.2、发明名称为“用于DMO的有效靶向的叶绿体转运肽及其用途”的发明专利申请的分案申请。
发明背景
本申请要求于2007年2月26日提交的美国临时专利申请60/891,675和2007年6月5日提交的美国专利申请1 1/758,659的优先权,本文完整引入其公开内容作为参考。
发明领域
本发明涉及植物生物技术领域。更具体的说,本发明涉及允许有效加工和定位植物中的麦草畏单加氧酶的叶绿体转运肽的鉴定和用途。
相关技术描述
DMO(麦草畏单加氧酶)在植物中催化除草剂麦草畏(3,6-二氯-邻茴香酸)降解为无毒的3,6-二氯水杨酸(3,6-DCSA),从而赋予除草剂耐受性。DMO的活性需要用于将电子从NADH向麦草畏转移的两种中间蛋白质:还原酶和铁氧还蛋白(美国专利7,022,896;Herman等人,2005)。然而,转基因植物中的麦草畏耐受性已通过单独用DMO转化得到证明,表明植物内源性的还原酶和铁氧还蛋白可以在电子转移中代替。参与电子转移的植物铁氧还蛋白位于质体中。因此,为了获得DMO的高效性能并因此获得提高的麦草畏耐受性,需要将DMO靶向至叶绿体。
在许多情况下,这种靶向可以通过被称作叶绿体转运肽(CTP)或者质体转运肽的N-末端延伸的存在而实现。如果表达的多肽将要在植物质体(如叶绿体)中区室化,细菌来源的染色体转基因必须具有与编码表达的多肽的序列相融合的编码CTP序列的序列。因此,将外源多肽定位到叶绿体中通常是借助于将编码CTP序列的多核苷酸序列可操作地连接到编码外源多肽的多核苷酸的5’区域上来完成的。CTP在向质体内转移过程中通过加工步骤被去除。但是,加工效率可能受CTP的氨基酸序列和肽氨基端附近的序列影响。
Weeks等人(美国专利7,022,896)描述了玉米cab-m7信号序列(参见,Becker等人,1992和PCT WO 97/41228;GenBank登记号X53398)和豌豆谷胱甘肽还原酶信号序列(Creissen等人,1992和PCT WO 97/41228)在将DMO靶向至植物质体方面的潜在用途,但是没有给出关于加工或者靶向效率的数据。含有包括豌豆核酮糖二磷酸羧化酶-加氧酶(Rubisco)小亚基编码序列的27个氨基酸序列的豌豆核酮糖二磷酸羧化酶-加氧酶小亚基(RbcS)CTP,也已经用于将DMO靶向至叶绿体(例如,美国临时申请60/811,152)。然而,已经在蛋白质印迹(Western blot)分析过程中发现,这种豌豆RbcS CTP产生一个正确加工的DMO蛋白质条带(~38 kDa),但是还产生一个与DMO和RbcS编码区的27个氨基酸相对应的较大的条带(~41kDa)。额外的氨基酸可能不利地影响DMO的活性。此外,由于DMO的不完全加工,转基因产品中额外的蛋白质造成监管方面的障碍,为了使产品在政府机构注册的目的,还需要在产品表征方面投入额外的努力,由此增加了产品注册的费用。因此,需要鉴定有效地产生正确加工的DMO的CTP,从而提供完全的DMO活性以及易于表征产品的优点。
发明概述
本发明的一方面涉及一种重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的核苷酸序列上的编码叶绿体转运肽的核苷酸序列,其中,该核苷酸序列编码包括选自SEQ ID NOs:1-11的序列的叶绿体转运肽。在某些实施方案中,重组DNA分子包括选自SEQ ID NOs:12-22的核苷酸序列。在某些实施方案中,重组DNA分子包括编码选自SEQ IDNOs:24、26、28、30、32、34、36、38和40的麦草畏单加氧酶的核苷酸序列。包括与在植物细胞中具有功能的启动子可操作地连接的DNA分子的DNA构建体也是本发明的一方面。
另一方面,本发明包括用DNA分子转化的植物细胞,所述DNA分子包括可操作地连接到编码麦草畏单加氧酶的核苷酸序列上的编码叶绿体转运肽的核苷酸序列,其中,叶绿体转运肽序列选自SEQ ID NOs:1-11。在某些实施方案中,重组DNA分子包括选自SEQ ID NOs:12-22的核苷酸序列。在某些实施方案中,DNA分子包括编码选自SEQ ID NOs:24、26、28、30、32、34、36、38和40的麦草畏单加氧酶的核苷酸序列,其中,DNA分子可操作地连接到在植物细胞中具有功能的启动子上。在特定的实施方案中,DNA分子包括选自SEQ ID NOs:23、25、27、29、31、33、35、37和39的核苷酸序列。
在某些实施方案中,植物细胞是双子叶植物细胞。在其它实施方案中,植物细胞是单子叶植物细胞。在特定的实施方案中,植物细胞是大豆、棉花、玉米或者油菜籽植物细胞。本发明也涉及包括这样的细胞的植物组织培养物,涉及包括这样的细胞的转基因种子和转基因植物。在某些实施方案中,转基因种子或者植物是双子叶种子或植物。在其它实施方案中,转基因种子或者植物是单子叶种子或者植物。转基因种子或者植物可以是大豆、棉花、玉米或者油菜籽种子或者植物。
本发明进一步涉及产生耐受麦草畏的植物的方法,包括:将重组DNA分子引入植物细胞中,并由其再生植物,所述DNA分子包括可操作地连接到编码麦草畏单加氧酶的核苷酸序列上的编码叶绿体转运肽的核苷酸序列,其中,编码叶绿体转运肽的序列选自SEQ ID NOs:12-22。在某些实施方案中,重组DNA分子包含编码选自SEQ ID NOs:24、26、28、30、32、34、36、38和40的麦草畏单加氧酶的核苷酸序列。DNA分子可以可操作地连接到在植物细胞中具有功能的启动子上。该方法可以进一步包括通过将亲本植株与它自身或者第二植株杂交产生耐受麦草畏的植物,其中亲本植株和/或第二植株包括所述DNA构建体,并且耐受麦草畏的植物从所述的亲本植株和/或第二植株遗传了该DNA构建体。
一种在植物细胞中表达麦草畏单加氧酶的方法是本发明的进一步的方面,该方法包括将选择的CTP可操作地连接到编码麦草畏单加氧酶的序列上。
另一方面,本发明涉及一种在农作物生长环境中控制杂草生长的方法,包括:种植这样的植物或其种子,并向农作物生长环境施用有效控制杂草生长的量的麦草畏除草剂。麦草畏除草剂可以自上方向农作物生长环境施用,由此麦草畏除草剂的量不损伤所述的植物或其种子,却损伤与该植物或其种子基因型相同但是缺乏构建体的植物或者种子。
本发明的进一步的方面涉及生产食物、饲料或者工业产品的方法,包括:
a)获得植物,该植物包括按照5’到3’的方向可操作地连接到编码叶绿体转运肽的核苷酸序列和编码麦草畏单加氧酶的核苷酸序列或其部分上的编码在植物细胞中具有功能的启动子的核苷酸序列;b)从所述植物或其部分制备食物、饲料、纤维或者工业产品。
在该方法的某些实施方案中,食物或者饲料是谷物、粗粉、油、淀粉、面粉或者蛋白质。在该方法的其它的实施方案中,工业产品是生物燃料、纤维、工业化学品、药物或者营养品。
针对出土前(pre emergence)施用麦草畏提供保护的、包括可操作地连接到编码麦草畏单加氧酶的DNA上的编码叶绿体转运肽的DNA的耐受麦草畏的种子,是本发明的进一步的方面。在某些实施方案中,耐受麦草畏的种子包括编码叶绿体转运肽的核苷酸序列,例如选自SEQ ID NOs:12-22的核苷酸序列。耐受麦草畏的种子可以进一步包括编码选自SEQ ID NOs:24、26、28、30、32、34、36、38和40的麦草畏单加氧酶的核苷酸序列。
本发明的另一方面涉及提高单子叶植物的直立能力(standability)的方法,包括:a)获得并培育通过将亲本植株与它自身或者第二植株杂交产生的植物,其中该亲本植株和/或第二植株包含所述DNA构建体并且耐受麦草畏的植物从所述的亲本植株和/或第二植株遗传了该DNA构建体;b)用麦草畏处理植物。在某些实施方案中,植物是玉米植物。在其它的实施方式中,可以测量包括支柱根的形状、数量、长度和/或结构、倒伏百分比和产量的与直立能力相关的参数。
本发明提供以下实施方案:
1.一种重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码选自SEQ ID NOs:1-11的序列。
2.如第1项所述的重组DNA分子,其中,所述编码叶绿体转运肽的DNA序列包括选自SEQID NOs:12-22的序列。
3.一种重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码选自拟南芥5-烯醇丙酮酰莽草酸-3-磷酸合酶CTP2叶绿体转运肽序列和豌豆核酮糖二磷酸羧化酶-加氧酶小亚基叶绿体转运肽序列的序列。
4.如第1项所述的重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码选自SEQ ID NOs:2-7的序列。
5.如第1项所述的重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码选自SEQ ID NO:2、SEQ ID NO:4和SEQ ID NO:5的序列。
6.如第1项所述的重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列包括选自SEQ ID NOs:13-18的序列。
7.如第1项所述的重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列包括选自SEQ ID NOs:13、15和16的序列。
8.如第1项所述的重组DNA分子,其中,所述编码麦草畏单加氧酶的DNA序列编码选自SEQ ID NOs:24、26、28、30、32、34、36、38和40的多肽。
9.如第8项所述的重组DNA分子,其中,所述DNA序列选自SEQ ID NOs:23、25、27、29、31、33、35、37和39。
10.一种DNA构建体,其包括与启动子可操作地连接的如第1项所述的DNA分子。
11.如第5项所述的DNA构建体,其中,所述启动子选自FMV35S启动子、At.ANT1启动子、FMV.35S-EF1a启动子、eIF4A10启动子、AGRtu.nos启动子、水稻胞质磷酸丙糖异构酶(OsTPI)启动子、水稻肌动蛋白15基因(OsAct15)启动子和γ-薏苡醇溶蛋白启动子。
12.如第10项所述的构建体,其中,所述启动子在植物细胞中是具有功能的。
13.用第10项所述的DNA构建体转化的植物细胞。
14.如第13项所述的细胞,其中,所述植物细胞是双子叶植物细胞。
15.如第13项所述的细胞,其中,所述植物细胞是单子叶植物细胞。
16.如第13项所述的细胞,其中,所述植物细胞是大豆、棉花、玉米或者油菜籽植物细胞。
17.一种植物组织培养物,其包含第13项所述的细胞。
18.如第17项所述的植物组织培养物,其包含双子叶植物细胞。
19.如第17项所述的植物组织培养物,其包含单子叶植物细胞。
20.如第17项所述的植物组织培养物,其包含大豆、棉花、玉米或者油菜籽植物细胞。
21.用第10项所述的DNA构建体转化的转基因植物。
22.如第21项所述的转基因植物,其中,所述植物是双子叶植物。
23.如第21项所述的转基因植物,其中,所述植物是单子叶植物。
24.如第21项所述的转基因植物,其中,所述植物是大豆、棉花、玉米或者油菜籽植物。
25.一种产生耐受麦草畏的植物的方法,包括将第10项所述的构建体引入植物细胞中,并由其再生包含第10项所述的构建体的植物。
26.如第25项所述的方法,进一步包括通过将亲本植株与它自身或者第二植株杂交产生耐受麦草畏的植物,其中,所述亲本植株和/或第二植株包含DNA构建体,并且耐受麦草畏的植物从所述的亲本植株和/或第二植株遗传了DNA构建体。
27.一种在植物细胞中表达麦草畏单加氧酶的方法,包括将选择的CTP可操作地连接到编码麦草畏单加氧酶的序列上。
28.一种在农作物生长环境中控制杂草生长的方法,包括第20项所述的植物或其种子,并向农作物生长环境施用有效控制杂草生长的量的麦草畏除草剂。
29.如第28项所述的方法,其中,所述麦草畏除草剂自上方向农作物生长环境施用。
30.如第28项所述的方法,其中,所述麦草畏除草剂的量不损伤第21项所述的植物或其种子,却损伤与第20项所述的植物的基因型相同但是缺乏第10项所述的构建体的植物。
31.一种生产食物、饲料或者工业产品的方法,包括:
a)获得第21项所述的植物或其部分;和
b)从该植物或者其部分制备食物、饲料、纤维或者工业产品。
32.如第31项所述的方法,其中,所述食物或者饲料是谷物、粗粉、油、淀粉、面粉或者蛋白质。
33.如第31项所述的方法,其中,所述工业产品是生物燃料、纤维、工业化学品、药物或者营养品。
34.一种针对出土前施用麦草畏提供保护的耐受麦草畏的种子,其包含可操作地连接到编码麦草畏单加氧酶的DNA上的编码叶绿体转运肽的DNA。
35.如第34项所述的耐受麦草畏的种子,其中,所述DNA编码选自SEQ ID NOs:1-11的叶绿体转运肽。
36.如第35项所述的耐受麦草畏的种子,其中,所述编码叶绿体转运肽的DNA包含选自SEQ ID NOs:12-22的序列。
37.如第34项所述的耐受麦草畏的种子,其中,所述DNA编码包含选自SEQID NOs:24、26、28、30、32、34、36、38和40的序列的麦草畏单加氧酶。
38.一种提高单子叶植物的直立能力的方法,包括:a)培育通过第26项所述的方法产生的植物或种子;和b)用麦草畏处理该植物或种子。
39.如第38项所述的方法,进一步包括:c)测量选自支柱根的数量、形状、长度或结构、倒伏百分比和产量的与直立能力相关的参数。
附图简述
图1:CTP-DMO构建体在DMO的正确加工和提供麦草畏耐受性中的应用。
发明详述
根据本发明,提供了用于在植物细胞中更有效地表达和向叶绿体转运麦草畏单加氧酶(DMO)多肽的组合物和方法。因此本发明的组合物和方法在增强植物和细胞对除草剂麦草畏的耐受性方面有用。尤其通过用叶绿体转运肽(CTP)将DMO靶向至叶绿体,可以实现提高的DMO表达和对麦草畏的耐受性。
然而,令人惊奇的是,本发明人已经发现某些CTP与DMO组合不能很好地发挥功能。例如,一些CTP不能导致足够的蛋白质表达。这包括蛋白质的不正确表达,以及改变大小的蛋白质的产生和不完全的体内活性。这会导致不完全的除草剂耐受性并且使注册审批变得复杂。本发明提供CTP,当该CTP与DMO联合使用时,提供意想不到的益处,包括但不限于:提高的转运至叶绿体的水平、在表达DMO的转基因植物中增强的除草剂耐受性、正确大小的蛋白质表达的理想水平和适当的翻译后修饰。当与DMO组合后提供意想不到的益处的CTP的一个例子是转运肽CTP2,包括SEQ ID NO:4或5的核酸,并且包括编码SEQ ID NOs:15和16的序列。在其它实施方案中,使用豌豆(Pisum sativum)核酮糖二磷酸羧化酶-加氧酶小亚基CTP编码序列,例如由SEQ ID NO:2所代表的或者编码SEQ ID NO:13。因此,包括可操作地连接到CTP2和/或豌豆核酮糖二磷酸羧化酶-加氧酶小亚基CTP转运肽编码序列上的DMO编码序列的DNA构建体构成本发明的一个方面,由其编码的蛋白质也构成本发明的一个方面。
嗜麦芽假单胞菌(Pseudomonas maltophilia)菌株DI6的麦草畏单加氧酶(Herman等人,2005;美国专利公开20030115626;GenBank登记AY786443,本文引入其编码DMO的序列作为参考)催化除草剂麦草畏的解毒。DMO是用于将麦草畏解毒成无毒的3,6-二氯水杨酸(3,6-DCSA)的三组分体系中的一部分,且如上所述需要还原酶和铁氧还蛋白功能来转移电子。由于参与电子转移的内源性植物铁氧还蛋白定位于质体中,为了获得DMO的有效活性和例如在双子叶植物中的麦草畏耐受性或者例如在单子叶植物中增强的麦草畏耐受性,优选DMO靶向至质体(例如叶绿体)。
测试叶绿体转运肽(CTP)在将DMO靶向至质体和DMO加工中的效率。与这些CTP有关的DMO的质体定位和加工从没有或部分到完全不等。发现只有一些CTP允许将DMO完全加工成正确的大小。因此,基于它的蛋白质或者核苷酸序列,任意给定的CTP提供完全和有效的DMO加工的能力是难以预料和出人意料的。
此外,也已经在拟南芥中发现,在没有合适的CTP的情况下,几乎没有或者没有DMO的表达与几乎没有或者没有麦草畏耐受性相关。这表明叶绿体靶向对于麦草畏的解毒进而对于耐受性是重要的。允许有效加工DMO的CTP在将DMO靶向至农作物的质体例如叶绿体方面是有用的,因此提供以下优势:完全的DMO活性和对麦草畏的较高的耐受性,以及易于表征产品和降低注册费用。
包括可操作地连接到编码麦草畏单加氧酶的DNA上的编码叶绿体转运肽的DNA的嵌合DNA分子可以通过本领域技术人员熟知的分子生物学方法制备(例如,Sambrook等人,1989)。本发明提供了可操作地连接到已知的编码DMO(包括表1中列出的那些)的DNA分子上的CTP,用于提高植物中DMO的表达。
来自在细胞核中编码的任何基因并且其产物可将多肽靶向至叶绿体的叶绿体转运肽,可以进行DMO有效表达的检测。可以分离或者合成叶绿体转运肽序列。为了在双子叶植物、单子叶植物或者这两者中表达,可以优化编码CTP的核苷酸序列。通过将每一个可操作地连接到DMO编码序列上,测试下面的转运肽:PsRbcS-衍生的CTPs(SEQ ID NO:1和2:豌豆核酮糖二磷酸羧化酶-加氧酶小亚基CTP;Coruzzi等人,1984);AtRbcS CTP(SEQ ID NO:3:拟南芥核酮糖二磷酸羧化酶-加氧酶小亚基1A CTP;CTP1;美国专利5,728,925);AtShkG CTP(SEQ ID NO:4:拟南芥5-烯醇丙酮酰莽草酸-3-磷酸合酶(EPSPS);CTP2;Klee等人,1987);AtShkGZm CTP(SEQ ID NO:5:CTP2合成的;为单子叶植物表达进行密码子优化;WO04009761的SEQ ID NO:14);PhShkG CTP(SEQ IDNO:6:矮牵牛(Petunia hybrida)EPSPS;CTP4;为单子叶植物表达进行密码子优化;Gasser等人,1988);TaWaxy CTP(SEQ ID NO:7:小麦(Triticum aestivum)结合颗粒的淀粉合酶CTP合成的,为玉米表达进行密码子优化:Clark等人,1991):OsWaxy CTP(SEQ ID NO:8:水稻(Oryza sativa)淀粉合酶CTP;Okagaki,1992);NtRbcS CTP(SEQ ID NO:9:烟草(Nicotiana tabacum)核酮糖1,5-二磷酸羧化酶小亚基叶绿体转运肽;Mazur,等人,1985);ZmAS CTP(SEQ ID NO:10:玉米(Zea mays)邻氨基苯甲酸合酶α2亚单位基因CTP;Gardiner等人,2004);和RgASCTP(SEQ ID NO:11:芸香(Ruta graveolens)邻氨基苯甲酸合酶CTP;Bohlmann,等人,1995)。编码SEQ ID NO:1-SEQ ID NO:11的核苷酸序列分别在SEQ ID NO:12-SEQ ID NO:22中给出。
可能有用的其它转运肽包括玉米cab-m7信号序列(Becker等人,1992;PCT WO 97/41228)和豌豆(Pisum sativum)谷胱甘肽还原酶信号序列(Creissen等人,1995;PCT WO 97/41228)。具有来自于基因编码区的额外氨基酸的CTP(所述氨基酸是该基因编码区的一部分或与它融合),例如AtRbcS CTP(其包含转运肽,成熟核酮糖二磷酸羧化酶-加氧酶蛋白的24个氨基酸,然后是转运肽的最后6个氨基酸的重复),可以被用来产生DMO。ZmAS CTP7也包含来自于基因编码区的额外18个氨基酸。其它CTP也可以用来产生DMO,它们具有来自于基因编码区的额外的氨基酸(例如27个氨基酸)(所述氨基酸是该基因编码区的一部分),例如PsRbcS CTP,紧接着是通过克隆方法引入的氨基酸(例如3个氨基酸)。具有较少的编码全长CTP的氨基酸(例如21个氨基酸)的CTP,例如RgAs CTP,也可以被用来产生DMO。优选地,使用编码全长CTP的核苷酸序列。可以包括一个或者多个核苷酸添加或者缺失,以便于CTP的克隆。这些添加或者缺失可以在其它表达元件和编码区的前面或者后面,导致一个或者多个编码氨基酸的修饰,例如在限制性酶识别位点处或者位于其附近。
在一个实施方案中,本发明涉及编码叶绿体转运肽的核酸序列,该叶绿体转运肽与SEQ ID NOs:1-11中的任意一个或者多个多肽序列至少具有70%的同一性,包括与这些序列有至少大约75%、80%、85%、90%、95%、97%、98%、99%或者更高的序列同一性,包括100%的同一性。在特定的实施方案中,所述核酸序列编码与SEQ ID NOs:1-11中的一个相同的叶绿体转运肽。在另一个实施方案中,编码CTP的核酸序列与SEQ ID NOs:12-22中的任意一个或者多个核酸序列具有至少70%的序列同一性,包括与这些序列中的一个或者多个至少有大约75%、80%、85%、90%、95%、97%、98%、99%或者更大的序列同一性,包括100%的同一性。这些序列和本文所述的任意其它序列的多肽或者多核苷酸比较和同一性的确定可以如本领域公知的那样进行,例如,使用MEGAlign(DNAStar,Inc.,1228S.Park St.,Madison,WI),采用缺省参数。这样的软件通过确定相似性或者同一性的程度,匹配相似的序列。
通过使用前序列可以将DMO靶向至其它细胞器例如线粒体,以利用存在于该细胞器中的铁氧还蛋白氧化还原***。或者,通过双靶向肽可以将DMO靶向至叶绿体和线粒体两者,以利用两个铁氧还蛋白氧化还原***,使工作更加有效。这样的元件是本领域的技术人员已知的。例如,线粒体前序列在Silva Filho等人,(1996)中有描述。编码双靶向肽序列的核酸序列可以从编码下述已知能被靶向至叶绿体和线粒体两者的蛋白质的核苷酸序列中被鉴定出来:Zn-MP(Moberg等人,2003)、谷胱甘肽还原酶(Rudhe等人,2002;Creissen等人,1995)和组氨酰-tRNA合成酶(Akashi等人,1998)。例如,如表1所示,在编码SEQ IDNOs:24、26、28、30、32、34、36、38、40多肽的序列中,发现可以用于这方面的DMO编码序列的例子。
表1.使用的DMO和DMO变体。
在某些实施方案中,编码麦单畏单加氧酶的核酸与编码SEQ IDNOs:24、26、28、30、32、34、36、38和40中任意一个多肽的序列具有至少70%的同一性,包括与这些序列具有至少大约75%、80%、85%、90%、95%、97%、98%、99%和更高的序列同一性。在某些实施方案中,核酸与SEQ ID NOs:23、25、27、29、31、33、35、37或39中的任意一个核酸序列具有至少70%的序列同一性,包括与这些序列中的一个具有至少大约75%、80%、85%、90%、95%、97%、98%、99%和更大的序列同一性。在进一步的实施方案中,麦草畏单加氧酶可以是任一个这样的序列的变体和/或可以是工程化的合成DMO分子,例如,如在2007年1月12日提交的名称为“DMO Methods AndCompositions”的美国临时申请60/884,854中描述的,本文特别引入其完整公开内容作为参考。
具有降解植物生长素样除草剂的能力的DMO的变体,以及草甘膦或者其它除草剂耐受基因,可以根据标准方法容易地制备,并测定其活性。也可以通过本领域已知的技术,例如,从合适的生物体中鉴定这些序列,所述生物体包括能够降解例如麦草畏的生物素样除草剂或者其它除草剂的细菌(美国专利5,445,962;Cork和Krueger,1991;Cork和Khalil,1995)。一种分离DMO或者其它序列的方法是通过例如与由来源生物体构建的文库进行核酸杂交,或者使用来自来源生物体的mRNA和基于公开的去饱和酶的引物进行RT-PCR。本发明因此包括在严格条件下与本文所述的DMO编码序列杂交的核酸的应用。本领域的技术人员理解通过增加盐浓度和降低温度可以提供严格性较低的条件。因此,杂交条件可以容易地控制,并且一般是根据预期的结果选择的方法。高严格性条件的一个例子是5X SSC、50%甲酰胺和42℃。通过在这样的条件下洗涤例如10分钟,可以除去那些在这些条件下不与特定靶序列杂交的序列。
变体也可以化学合成,例如,根据本领域众所周知的技术,利用已知的DMO多核苷酸序列合成。例如,DNA序列可以通过亚磷酰胺化学法(phosphoroamidite)在自动DNA合成仪上合成。化学合成有许多优势。特别是,化学合成是合乎需要的,因为将要表达DNA序列的宿主所偏好的密码子可以用来优化表达。并非所有的密码子都需要改变来获得改进的表达,但是优选地至少将那些在宿主中很少用到的密码子改变为宿主偏好的密码子。通过改变超过大约50%,最优选至少大约80%的密码子成为宿主偏好的密码子,可以得到高水平的表达。许多宿主细胞的密码子偏好是已知的(例如,PCT WO 97/31115;PCT WO 97/11086;EP 646643;EP 553494;和美国专利:5,689,052;5,567,862;5,567,600;5,552,299和5,017,692)。可以通过本领域已知的方法,推导出其它宿主细胞的密码子偏好。此外,利用化学合成,可以容易地改变DNA分子的序列或者其编码的蛋白质,例如用来优化表达(例如,消除干扰转录或翻译的mRNA二级结构)、在方便的位点加入独特的限制性位点、以及删除蛋白酶酶切位点。
在根据需要保留或者改变酶活性的同时,可以对蛋白质的多肽序列,如本文提供的DMO序列,进行修饰和改变。在2007年1月12日提交的美国临时申请60/884,854中提供了产生DMO序列的示例性的方法。下面是基于改变蛋白质的氨基酸来产生相当的或甚至改进的、修饰的多肽和相应的编码序列的讨论。已知,例如,在蛋白质结构中某些氨基酸可以被其它氨基酸代替,却不会明显损失与底物分子上诸如结合位点的结构相互结合的能力。因为蛋白质的相互作用能力和性质限定了蛋白质的生物学功能活性,所以可以在蛋白质序列中,当然也可以在其DNA编码序列中,进行某些氨基酸序列置换,而仍然获得具有相似性质的蛋白质。因此设想:可以对本文描述的DMO肽序列或者其它除草剂耐受性多肽和相应的DNA编码序列进行各种改变,而不会明显减弱它们的生物学效用或者活性。
进行这样的改变时,需要考虑氨基酸的亲水指数。本领域中一般理解亲水氨基酸指数在赋予蛋白质相互作用生物学功能方面的重要性(Kyte等人,1982)。普遍认为,氨基酸的相对亲水特性对产生的蛋白质的二级结构具有贡献,而蛋白质的二级结构又限定了蛋白质与例如酶、底物、受体、DNA、抗体、抗原等其它分子的相互作用。基于它们的疏水性和电荷特性,每种氨基酸已被分配了亲水指数(Kyte等人,1982),这些是:异亮氨酸(+4.5);缬氨酸(+4.2);亮氨酸(+3.8);苯丙氨酸(+2.8);半胱氨酸/胱氨酸(+2.5);甲硫氨酸(+1.9);丙氨酸(+1.8);甘氨酸(-0.4);苏氨酸(-0.7);丝氨酸(-0.8);色氨酸(-0.9);酪氨酸(-1.3);脯氨酸(-1.6);组氨酸(-3.2);谷氨酸(-3.5);谷氨酰胺(-3.5);天冬氨酸(-3.5);天冬酰胺(-3.5);赖氨酸(-3.9)和精氨酸(-4.5)。
本领域已知,氨基酸可以被其它具有相似亲水指数或者得分的氨基酸代替,而仍然产生具有相似生物学活性的蛋白质,即,仍然获得生物学功能相当的蛋白质。进行这样的转变时,优选其亲水指数在±2之内的氨基酸置换,尤其优选在±1之内的氨基酸置换,甚至更优选在±0.5之内的氨基酸置换。
本领域中也理解在亲水性的基础上可以有效进行相似氨基酸的置换。美国专利4,554,101提出,受到邻近氨基酸亲水性控制的蛋白质的最大局部平均亲水性与蛋白质的生物学性质相关。如美国专利4,554,101所述,下面的亲水性值被分配给氨基酸残基:精氨酸(+3.0);赖氨酸(+3.0);天冬氨酸(+3.0±1);谷氨酸(+3.0±1);丝氨酸(+0.3);天冬酰胺(+0.2);谷氨酰胺(+0.2);甘氨酸(0);苏氨酸(-0.4);脯氨酸(-0.5±1);丙氨酸(-0.5);组氨酸(-0.5);半胱氨酸(-1.0);甲硫氨酸(-1.3);缬氨酸(-1.5);亮氨酸(-1.8);异亮氨酸(-1.8);酪氨酸(-2.3);苯丙氨酸(-2.5);色氨酸(-3.4)。应当理解:氨基酸可以被另一种具有相似亲水性值的氨基酸代替,而仍然获得生物学相当的蛋白质。在这样的改变中,优选其亲水性值在±2之内的氨基酸置换,尤其优选在±1之内的氨基酸置换,甚至更优选在±0.5之内的氨基酸置换。将这些和各种上述特征考虑在内的示例性的置换是本领域的技术人员熟知的,且包括:精氨酸和赖氨酸,谷氨酸和天冬氨酸,丝氨酸和苏氨酸,谷氨酰胺和天冬酰胺,及缬氨酸、亮氨酸和异亮氨酸。
包括可操作地连接到DMO序列上的CTP序列的DNA构建体可以通过被可操作地连接到在植物中具有功能的启动子上而在诸如原生质体、瞬时或者稳定转化的植物细胞等测试***中表达。描述这样的启动子的例子包括美国专利6,437,217(玉米RS81启动子)、美国专利5,641,876(水稻肌动蛋白启动子;OsAct1)、美国专利6,426,446(玉米RS324启动子)、美国专利6,429,362(玉米PR-1启动子)、美国专利6,232,526(玉米A3启动子)、美国专利6,177,611(组成型玉米启动子)、美国专利5,322,938、5,352,605、5,359,142和5,530,196(35S启动子)、美国专利6,433,252(玉米L3油质蛋白启动子)、美国专利6,429,357(水稻肌动蛋白2启动子以及水稻肌动蛋白2内含子)、美国专利5,837,848(根特异性启动子)、美国专利6,294,714(光诱导型启动子)、美国专利6,140,078(盐诱导型启动子)、美国专利6,252,138(病原体诱导型启动子)、美国专利6,175,060(磷缺乏诱导型启动子)、美国专利6,388,170(例如,PC1SV启动子)、SEQ ID NO:41的PC1SV启动子、美国专利6,635,806(γ-薏苡醇溶蛋白(coixin)启动子)和美国专利7,151,204(玉米叶绿体醛缩酶启动子)。可以使用的其它启动子是胭脂氨酸合酶(NOS)启动子(Ebert等人,1987)、章鱼氨酸合酶(OCS)启动子(其在根癌土壤杆菌(Agrobacterium tumefaciens)的肿瘤诱导的质粒中携带)、花椰菜花叶病毒启动子例如花椰菜花叶病毒(CaMV)19S启动子(Lawton等人,1987)、CaMV 35S启动子(Odell等人,1985)、玄参花叶病毒35S-启动子(Walker等人,1987)、蔗糖合酶启动子(Yang等人,1990)、R基因复合体启动子(Chandler等人,1989)和叶绿素a/b结合蛋白基因启动子等。在本发明中,具有增强子序列的CaMV35S(e35S;美国专利5,322,938;5,352,605;5,359,142和5,530,196)、FMV35S(美国专利6,051,753;5,378,619)、花生褪绿条纹花椰菜花叶病毒(PC1SV;美国专利5,850,019)、At.Act 7(登记号#U27811)、At.ANT1(美国专利申请20060236420)、FMV.35S-EF1a(美国专利申请20050022261)、eIF4A10(登记号#X79008)和AGRtu.nos(GenBank登记号V00087;Depicker等人,1982;Bevan等人,1983)、水稻胞质磷酸丙糖异构酶(OsTPI;美国专利7,132,528)和水稻肌动蛋白15基因(OsAct15;美国专利申请2006-0162010)启动子可能特别有用。
作为翻译前导序列起作用的5’UTR是位于基因的启动子序列和编码序列之间的DNA遗传元件,可以被包括在启动子和CTP-DMO序列之间。翻译前导序列存在于翻译启动序列上游的完全加工的mRNA之中。翻译前导序列可以影响初级转录物向mRNA的加工、mRNA稳定性或者翻译效率。翻译前导序列的例子包括玉米和矮牵牛花热休克蛋白前导序列(美国专利5,362,865)、植物病毒外壳蛋白前导序列、植物核酮糖二磷酸羧化酶-加氧酶前导序列、GmHsp(美国专利5,659,122)、PhDnaK(美国专利5,362,865)、AtAnt1、TEV(Carrington和Freed,1990)和AGRtunos(GenBank登记号V00087;Bevan等人,1983)等(Turner和Foster,1995)。在本发明中,可能尤其有益的5’UTR是GmHsp(美国专利5,659,122)、PhDnaK(美国专利5,362,865)、AtAnt1、TEV(Carrington和Freed,1990)、OsAct1(美国专利5641876)、OsTPI(美国专利7,132,528)、OsAct15(美国公开20060162010)和AGRtunos(GenBank登记号V00087;Bevan等人,1983)。
3’非翻译序列、3’转录终止区或者多腺苷酸化区域意思是连接到并位于结构多核苷酸分子下游的DNA分子,并且包括提供能够影响转录、mRNA加工或者基因表达的多腺苷酸化信号和其它调控信号的多核苷酸。多腺苷酸化信号在植物中发挥功能,以导致向mRNA前体的3’末端增加多聚腺苷酸。多腺苷酸化序列可以来源于天然基因、多种植物基因或者T-DNA基因。这些序列可以包含于CTP-DMO序列的下游。3’转录终止区的例子是胭脂氨酸合酶3’区(nos 3’;Fraley等人,1983)。举例说明了不同的3’非翻译区的应用(Ingelbrecht等人,1989)。来自豌豆RbcS2基因(Ps.RbcS2-E9;Coruzzi等人,1984)、AGRtu.nos(Genbank登记号E01312)、E6(登记号#U30508)和TaHsp17(小麦低分子量热休克蛋白基因;登记号#X13431)的多腺苷酸化分子特别有利地用于本发明。
除了上述的表达元件以外,为了尤其在单子叶植物中表达编码区,在启动子和3’UTR之间可能需要内含子。“内含子”指的是多核苷酸分子,其可以从基因的基因组拷贝的间插序列分离或者鉴定,而且一般可以被定义为在翻译前mRNA加工过程中被剪切掉的区域。或者,内含子可以合成产生。内含子本身可以包含影响可操作地连接的基因的转录的亚元件例如顺式元件或者增强子区域。“植物内含子”是在植物细胞中发挥功能的天然或者非天然的内含子。植物内含子可以用作调控元件来调节可操作地连接的一种或多种基因的表达。转化构建体中的多核苷酸分子序列可以包含内含子。相对于可以转录的多核苷酸序列,内含子可以是异源的。内含子的例子包括玉米肌动蛋白内含子(美国专利5641876)、玉米HSP70内含子(ZmHSP70;美国专利5,859,347;美国专利5,424,412)和水稻TPI内含子(OsTPI;美国专利7,132,528),并且在实施本发明中是有益的。
可以通过植物组织培养和转化领域的技术人员所知的方法,在如原生质体或者瞬时或稳定转化的单子叶或双子叶植物细胞的测试***中测试CTP-DMO构建体是否提供DMO的正确加工。按照本发明,可以使用本领域中已知的任一种将转基因构建体导入植物的技术(参见,例如,Miki等人,1993)。据信,合适的植物转化方法实际上包括任一种可将DNA导入细胞中的方法,如美国专利5,384,253所描述的电穿孔法;美国专利5,015,580、5,550,318、5,538,880、6,160,208、6,399,861和6,403,865所描述的微粒轰击法;美国专利5,635,055、5,824,877、5,591,616、5,981,840和6,384,301所描述的土壤杆菌介导转化法;和美国专利5,508,184描述的原生质体转化法。通过应用如这些方法的技术,实际上任何植物种的细胞都可以稳定地转化,而且这些细胞可以发育成转基因植物。美国专利5,846,797、5,159,135、5,004,863和6,624,344公开了在棉花转化中特别有用的技术。例如,在美国专利5,750,871中特别公开了转化芸苔属(Brassica)植物的技术;例如,在Zhang等人,1999、美国专利6,384,301和US 7,002,058中公开了转化大豆的技术。在WO9506722中公开了转化玉米的技术。可以用于本发明的植物的一些非限制性例子包括苜蓿、大麦、豆类、甜菜、花茎甘蓝、卷心菜、胡萝卜、油菜、花椰菜、芹菜、大白菜、玉米、棉花、黄瓜、干豆、茄子、茴香、青刀豆、葫芦、韭菜、莴苣、甜瓜、燕麦、黄秋葵、洋葱、豌豆、胡椒、南瓜、花生、马铃薯、南瓜、萝卜、水稻、高粱、大豆、菠菜、倭瓜、甜玉米、甜菜、向日葵、西红柿、西瓜和小麦。
实现将外源DNA导入接受体细胞后,产生转基因植物的下一步一般涉及鉴定用于进一步的培养和植物再生的转化的细胞。为了提高鉴定转化体的能力,可能需要与按照本发明制备的转化载体一起使用可选择或可筛选的标记基因。在这种情况下,一般接下来通过将细胞暴露于一种或多种选择剂检测可能转化的细胞群,或者针对需要的标记基因的性状筛选细胞。
在暴露于选择剂后存活的细胞,或者在筛选试验中评分为阳性的细胞,可以在支持植物再生的培养基上继续培养。任何合适的植物组织培养基,例如,MS或者N6培养基(Murashige和Skoog,1962;Chu等人,1975),可以通过包含如生长调节剂的其它物质进行改变。可以在含有生长调节剂的基础培养基上维持组织,直到获得足够的组织来开始植物再生工作,或者进行反复几轮人工选择,直到组织的形态适合再生,典型地至少两周,然后转移到有助于幼苗形成的培养基上。定期转移培养物,直到发生足够的幼苗形成。一旦幼苗形成,就将它们转移到有助于根形成的培养基上。一旦形成足够的根,就可以将植物转移到土壤中进一步生长和成熟。
为了证实外源DNA或者“转基因”在再生植物中的存在,可以进行多种试验。这样的试验包括,例如,“分子生物学”试验,如DNA印迹法和RNA印迹法和PCRTM;“生物化学”试验,如检测蛋白质产物的存在,例如,通过免疫学方法(ELISA和蛋白质印迹法)或者通过酶功能;植物部分试验,如叶或者根试验;以及,通过分析整个再生植物的表型。
一旦转基因被导入植物中,就可以通过杂交将该基因导入到任何与该第一植物性相容的植物中,而根本不需要直接转化第二种植物。因此,本文中使用的术语“后代”指的是按照本发明制备的亲本植株的任何一代后代,其中,该后代包含按照本发明制备的选择的DNA构建体。因此,“转基因植物”可以是任何一代。如本文公开的,“杂交”一种植物以提供相对于初始植物系具有一个或者多个添加的转基因或者等位基因的植物系,被定义为通过将初始系与包含本发明的转基因或者等位基因的供体植物系杂交从而导致特定的序列被导入植物系中的技术。为了实现这一点,例如,可以进行如下步骤:(a)种植第一(初始系)和第二(包含需要的转基因或者等位基因的供体植物系)亲本植株的种子;(b)将第一和第二亲本植株的种子培育成有花的植物;(c)用第二亲本植物的花粉对第一亲本植物的花授粉;和(d)收获具有已受精的花的亲本植物上产生的种子。
可以测试稳定转化的植物组织和植物是否通过DMO蛋白质的正确加工提供麦草畏耐受性。在农作物植物中提供麦草畏耐受性可以用来设计控制生长环境中的杂草生长的方法,包括:向农作物生长环境施用有效控制杂草生长的量的麦草畏除草剂。麦草畏除草剂以一定的量自上方向农作物生长环境施用,所述麦草畏除草剂的量不损伤用CTP-DMO构建体转化的农作物植物或种子,却损伤具有同样的基因型但是缺乏CTP-DMO构建体的农作物植物。
基于公开的内容,用于本发明的除草剂组合物的制备对于本领域的技术人员来说是显而易见的。这样的可商购的组合物除了活性成分外,典型地还包含如表面活性剂、固体或液体载体、溶剂和粘合剂等组分。可用于向植物施用的表面活性剂的例子包括以下物质的碱金属盐、碱土金属盐或铵盐:芳香族磺酸类,例如,木素磺酸、苯酚磺酸、萘磺酸和二丁基萘磺酸,和脂肪酸、芳基磺酸酯、烷基醚、月桂基醚、脂肪醇硫酸盐和脂肪醇二醇醚硫酸盐,磺化萘及其衍生物与甲醛的缩合物、萘或萘磺酸与苯酚和甲醛的缩合物、苯酚或苯酚磺酸与甲醛的缩合物、苯酚与甲醛和亚硫酸钠的缩合物、聚氧乙烯辛基苯基醚、乙氧基化的异辛基-、辛基-或壬基苯酚、三丁基苯基聚二醇醚、烷基芳基聚醚醇、异十三烷醇、乙氧基化的蓖麻油、乙氧基化的三芳基苯酚、磷酸化的三芳基苯酚乙氧基化物的盐、月桂醇聚二醇醚乙酸酯、山梨醇酯、木质素-亚硫酸盐废液或甲基纤维素,或它们的混合物。表面活性剂应用的常见实例是大约0.25重量%-1.0重量%,更常用地为大约0.25重量%-0.5重量%。
向植物施用的组合物可以是固体或液体。使用固体组合物时,可能需要包含一种或多种载体材料和活性化合物。载体的例子包括矿物土,如硅石、硅胶、硅酸盐、滑石、高岭土、镁质粘土(attaclay)、石灰石(limestone)、白垩(chalk)、黄土(loess)、粘土、白云石(dolomite)、硅藻土(diatomaceous earth)、硫酸钙、硫酸镁、氧化镁、磨碎的合成材料,肥料,如硫酸铵、磷酸铵、硝酸铵、硫脲和脲,植物来源的产品,如谷物粗粉、树皮粗粉、木粉和坚果壳粗粉、纤维素粉、硅镁土(attapulgites)、蒙脱石(montmorillonites)、云母(mica)、蛭石(vermiculites)、合成的二氧化硅和合成的硅酸钙,或它们的混合物。
对于液体溶液,可以包含水溶性的化合物或盐,如硫酸钠、硫酸钾、氯化钠、氯化钾、醋酸钠、硫酸氢铵、氯化铵、醋酸铵、甲酸铵、草酸铵、碳酸铵、碳酸氢铵、硫代硫酸铵、二磷酸氢铵、磷酸二氢铵、磷酸氢铵钠、硫氰酸铵、氨基磺酸铵或氨基甲酸铵。
除草剂组合物中的其它示例性的成分包括:粘合剂,如聚乙烯吡咯烷酮、聚乙烯醇、部分水解的聚醋酸乙烯酯、羧甲基纤维素、淀粉、乙烯吡咯烷酮/醋酸乙烯酯共聚物和聚醋酸乙烯酯或它们的混合物;润滑剂,如硬脂酸镁、硬脂酸钠、滑石或聚乙二醇或它们的混合物;防泡剂,如乳化硅油、长链醇、磷酸酯、乙炔二醇、脂肪酸或有机氟化合物;和螯合剂,如:乙二胺四乙酸(EDTA)的盐、三次氮基三乙酸的盐或聚磷酸的盐或它们的混合物。
可以使用从大约2.5g/ha到大约10,080g/ha的麦草畏,包括大约2.5g/ha-大约5,040g/ha、大约5g/ha-大约2,020g/ha、大约10g/a-大约820g/h和大约50g/ha-大约1,000g/ha、大约100g/ha-大约800g/ha和大约250g/ha-大约800g/ha。
CTP-DMO构建体可以连接到包含用于可筛选/可评分/可选择的标记和/或赋予另一种需要的性状的遗传元件的一个或者多个多核苷酸分子上。通常用来筛选推测转化的细胞的基因包括β-葡糖醛酸糖苷酶(GUS)、β-半乳糖苷酶、萤光素酶和氯霉素乙酰转移酶(Jefferson,1987;Teeri等人,1989;Koncz等人1987;De Block等人,1984)、绿色荧光蛋白(GFP)(Chalfie等人,1994;Haseloff和Amos,1995;和PCT申请WO97/41228)。例如,在Miki和McHugh,2004中描述了选择性标记基因的非限制例子。
赋予另一种需要的性状的核苷酸分子包括,但不限于:提供需要的与植物形态、生理、生长和发育、产量、营养强化、抗病性或抗虫性、或环境或化学耐受性有关的特性的基因,还可以包括遗传元件,包括:除草剂抗性(美国专利6,803,501;6,448,476;6,248,876;6,225,114;6,107,549;5,866,775;5,804,425;5,633,435;5,463,175)、增加的产量(美国专利RE38,446;6,716,474;6,663,906;6,476,295;6,441,277;6,423,828;6,399,330;6,372,211;6,235,971;6,222,098;5,716,837)、害虫控制(美国专利6,809,078;6,713,063;6,686,452;6,657,046;6,645,497;6,642,030;6,639,054;6,620,988;6,593,293;6,555,655;6,538,109;6,537,756;6,521,442;6,501,009;6,468,523;6,326,351;6,313,378;6,284,949;6,281,016;6,248,536;6,242,241;6,221,649;6,177,615;6,156,573;6,153,814;6,110,464;6,093,695;6,063,756;6,063,597;6,023,013;5,959,091;5,942,664;5,942,658,5,880,275;5,763,245;5,763,241)、真菌病抗性(美国专利6,653,280;6,573,361;6,506,962;6,316,407;6,215,048;5,516,671;5,773,696;6,121,436;6,316,407;6,506,962)、病毒抗性(美国专利6,617,496;6,608,241;6,015,940;6,013,864;5,850,023;5,304,730)、线虫抗性(美国专利6,228,992)、细菌性疾病抗性(美国专利5,516,671)、植物生长发育(美国专利6,723,897;6,518,488)、淀粉生产(美国专利6,538,181;6,538,179;6,538,178;5,750,876;6,476,295)、改变的油的生产(美国专利6,444,876;6,426,447;6,380,462)、高油产量(美国专利6,495,739;5,608,149;6,483,008;6,476,295)、改变的脂肪酸含量(美国专利6,828,475;6,822,141;6,770,465;6,706,950;6,660,849;6,596,538;6,589,767;6,537,750;6,489,461;6,459,018)、高蛋白质产量(美国专利6,380,466)、果实成熟(美国专利5,512,466)、加强的动物和人的营养(美国专利6,723,837;6,653,530;6,541,259;5,985,605;6,171,640)、生物聚合物(美国专利RE37,543;6,228,623;5,958,745和美国专利公开US20030028917)、环境应激耐受性(美国专利6,072,103)、药物肽和可分泌的肽(美国专利6,812,379;6,774,283;6,140,075;6,080,560)、改进的加工性状(美国专利6,476,295)、提高的可消化性(美国专利6,531,648)、低水平的棉子糖(美国专利6,166,292)、工业酶生产(美国专利5,543,576)、改进的香味(美国专利6,011,199)、固氮(美国专利5,229,114)、杂交种子生产(美国专利5,689,041)、纤维生产(美国专利6,576,818;6,271,443;5,981,834;5,869,720)和生物燃料生产(美国专利5,998,700)。这些或者其它的遗传元件、方法和转基因中的任一种可以用于本发明,是本领域的技术人员基于本公开内容可以理解的。
另外,一个或者多个连接到CTP-DMO构建体上的多核苷酸分子通过编码导致内源基因表达的靶向抑制(例如经过反义、抑制性RNA(RNAi)或者共抑制介导的机制)的RNA分子,可以实现上面提到的植物特性或者表型。RNA也可以是催化性的RNA分子(即核酶),设计用来切割需要的内源性mRNA产物。因此,任何编码影响感兴趣的表型或者形态学改变的转录RNA分子的多核苷酸分子可以用于实践本发明。
本发明也公开了生产食物、饲料或工业产品的方法,包括包含CTP-DMO构建体的植物或者这种植物的一部分,以及从植物或者其部分制备食物、饲料、纤维或工业产品,其中,食物或饲料是谷物、粗粉、油、淀粉、面粉或蛋白质,且工业产品是生物燃料、纤维、工业化学品、药物或营养品。
本发明的另一方面涉及改进单子叶植物的直立能力的方法,包括:a)获得并种植通过将亲本植株与它自身或者第二植株杂交产生的植物,其中亲本植株和/或第二植株包含所述DNA构建体,并且耐受麦草畏的植物从所述的亲本植株和/或第二植株遗传了该DNA构建体;和b)用麦草畏处理该植物。可以测量包括支柱根的数量、形状、长度或结构、倒伏百分比和产量的与直立能力相关的参数。在某些实施方案中,植物是玉米植物。
实施例
下述这些实施例用来说明本发明的实施方案。本领域的技术人员应该理解:下面的实施例中公开的技术代表了本发明人发现的、在本发明的实践中良好地发挥作用的技术。然而,本领域的技术人员根据本发明的公开内容应该理解,可以对公开的特定的实施方案进行许多改变,仍然获得相同或类似的结果,而没有偏离本发明的概念、精神和范围。更具体地说,显然某些化学和生理学相关的试剂可以代替本文中所述的试剂,而将获得相同或相似的结果。所有这些对于本领域技术人员明显的类似的代替和修改,被认为是在所附的权利要求书限定的本发明的精神、范围和概念之内。
实施例1
用于转化的CTP-DMO构建体的制备
按照标准方法(例如Sambrook等人,1989)制备表2中所示的DNA构建体,该DNA构建体在植物启动子和多腺苷酸化信号序列之间包含与DMO基因或其变体可操作地连接的CTP。如下所述,在玉米原生质体***或在稳定转化的拟南芥或者大豆植物中测试这些构建体。
实施例2
玉米原生质体中的CTP-DMO构建体的分析
从12天的黄化幼苗(来自LH200x LH5杂交)制备玉米叶叶肉原生质体。将第二叶的中间部分(大约6cm长)用刀片切割成0.5mm的条,并且在真空过滤30分钟后,在烧瓶中用含有2%(w/v)纤维素酶RS、0.3%(w/v)离析酶R10(都来自Karlan Research Products Corp,SantaRosa,CA)、0.6M甘露醇、10mM MES(pH 5.7)和1mM CaCl2的酶溶液在23℃消化不超过2小时。来自过滤的和消化的叶组织的原生质体通过用手摇动烧瓶5分钟释放,并通过60-μm尼龙网过滤分离。原生质体通过以150g离心2分钟收集,用0.6M冷甘露醇溶液洗涤一次,离心,以2×106/mL再悬浮于冷0.6M甘露醇中。然后使用聚乙二醇(PEG),以12.5μgDNA转化原生质体,并在室温下孵育16-20小时。
原生质体存储于-80℃,直到通过蛋白质印迹法(western blot)分析。原生质体在冰上融化,并将1-3倍体积的含有5.0%β-ME的2xLaemmli样品缓冲液/染料(BioRad)加到原生质体。然后将原生质体蛋白质样本的等分试样加热到大约100℃保持5分钟,并加样到预制的Tris-HCL 10%聚丙烯酰胺凝胶上。在大约80-100Amps的恒定电流下电泳大约35分钟。然后将蛋白质在100V恒定电压下从凝胶电转移至0.2微米的硝酸纤维素膜上进行1-3小时。膜用TBST中的5%(w/v)奶粉在室温下封闭1小时或者在4℃下封闭过夜。用在TBST中1∶200稀释的山羊抗-DMO抗体探测膜1小时。使用TBS冲洗3次,每次5分钟,以除去过量的抗体。用在含0.5%(w/v)奶粉的TBST中以1∶7,500比例稀释的过氧化物酶偶联的兔抗山羊IgG(Sigma,St.Louis,MO)探测膜1小时。用TBST冲洗3次,每次5分钟,以除去过量的过氧化物酶偶联物。除了那些特别指出的以外,所有的程序,包括封闭和所有的其它孵育,在室温下进行。使用ECL检测***(Amersham Biosciences,Piscataway,NJ)显示免疫活性的条带,并在Kodak BioMaxTM MS胶片上曝光。适当大小的免疫活性条带的存在表明DMO的正确加工和定位(表1)。因此,例如,使用可操作地连接到DMO上并转化到玉米原生质体中的CTP4在蛋白质印迹分析后产生38kDa的免疫活性条带。
实施例3
各种CTP-DMO构建体在拟南芥中的测试
按照Clough和Bent(1998)开发的方法,转化拟南芥哥伦比亚生态型植物。将通过这种方法获得的种子接种到含有0.5、1.0到2.0或4.0毫克/升的多种浓度麦草畏的植物选择培养基上。将这些板在4℃孵育48小时,然后转移到设定为23.5℃、具有16小时光周期的Percival孵箱中。用CTP-DMO构建体转化的种子在含有麦草畏的培养基上生长成植物,并长出初生叶和次生叶,而未转化的种子和阴性分离子(segregants)死亡或者没有长出初生叶和次生叶。通过PCR试验检测为3’UTR阳性的转基因植物用于进一步分析。
来自转基因拟南芥植物的3-5个叶穿孔用于蛋白质印迹分析。在Harbil paint振摇器中用500-1000μl PSBT和4个玻璃珠进行蛋白质提取3分钟。在4℃下以3000rpm旋转样本3分钟。向上清液的等分试样中加入等体积的含有5.0%β-ME的2x Laemmli样品缓冲液/染料(cat.No.161-0737 BioRad)。蛋白质印迹分析的其余步骤与实施例2中一样。正确大小的免疫活性条带的存在表明DMO的正确加工和定位(表2)。例如,如表2所示,在用pMON73749或者pMON73725转化拟南芥后可见的条带的比较中,使用缺少来自豌豆核酮糖二磷酸羧化酶-加氧酶的27个氨基酸的编码序列的RbcSnoc-CTP产生定位于叶绿体的正确加工的DMO,而使用包含27个氨基酸的编码序列的RbcS CTP则产生2个免疫活性条带。
实施例4
CTP-DMO构建体在大豆中的测试
使用标准程序通过土壤杆菌介导的大豆转化(例如,Zhang等人,1999;US 7,002,058)获得转基因大豆(例如,cvs.Thorne,NE3001和A3525)植物。来自转基因大豆植物的3-5个叶穿孔用于蛋白质印迹分析。在Harbil paint振摇器中用500-1000μl PSBT和4个玻璃珠进行蛋白质提取3分钟。在4℃下以3000rpm旋转样本3分钟。向上清液的等分试样中加入等体积的2x Laemmli样品缓冲液/染料(BioRad)/5.0%β-ME。蛋白质印迹分析的其余步骤与实施例2中一样。正确大小的免疫活性条带的存在表明DMO的正确加工和定位(表2)。
用构建体(其编码连接到附着有核酮糖二磷酸羧化酶-加氧酶编码区的另外24个氨基酸和由于引入限制性酶识别位点而添加的3个氨基酸的豌豆核酮糖二磷酸羧化酶-加氧酶转运肽上的DMO)转化的大豆植物,当在出土前阶段用0.51b的麦草畏处理接着在出土后(V6)阶段用21b麦草畏处理时,显示17-20%的损伤率。将其与用编码只连接到豌豆核酮糖二磷酸羧化酶-加氧酶转运肽上的DMO的构建体转化的大豆植物进行比较,后者显示大约12%的损伤率。这些结果表明使用没有额外氨基酸的转运肽导致单一DMO活性的产生(而不是多个部分或不同加工的多肽)和对麦草畏的较高的耐受性。单一形式的酶的产生也导致易于表征产品和降低注册费用。
实施例5
DMO的高效生产和较高的麦草畏耐受性需要CTP
用如实施例3所述的几种构建体(图1)转化拟南芥哥伦比亚生态型植物。在包含从0.5、1.0到2.0毫克/升的各种浓度的麦草畏的植物组织培养基上选择转化的种子。用CTP-DMO构建体转化的种子在含有麦草畏的培养基上生长成植物,并长出初生叶和次生叶,而未转化的种子和阴性分离子死亡或者没有长出初生叶和次生叶。生长并检测为DMO基因阳性的转基因植物用于进一步分析。
如图1所示,用不含CTP的构建体转化的植物几乎没有或者没有表现出对麦草畏的耐受性。当在出土前阶段用0.5lb/a的麦草畏处理接着在出土后(V6)阶段用21b/a的麦草畏处理时,用编码没有连接到CTP上的DMO的DNA构建体转化的大豆植物没有显示出土前耐受性,而用其中DMO连接到CTP上的构建体转化的植物显示出土前和出土后对麦草畏的耐受性。
实施例6
耐受麦草畏的转基因玉米植物的产生
为了测试DMO基因在向单子叶植物提供麦草畏耐受性方面的用途,在植物基因表达元件如启动子(例如,PC1SV、e35S、OsAct1、OsTPI、OsAct15)和内含子(例如,OsAct1、OsAct15、OsTPI、ZmHSP70)的控制下,产生了转基因玉米植物,其包含DMO基因(例如,SEQ ID NOS:29、33、35、37、39),具有或者不具有转运肽(例如,TaWaxy、CTP1、CTP2合成、CTP4)。该表达元件包含来自水稻肌动蛋白1基因的第一内含子和侧翼UTR外显子序列,还在5’端包含外显子1的12nt和在3’端包含外显子2的7nt,还有3’UTR(例如,TaHsp17)。
转基因玉米植物基本上通过美国专利申请20040244075所述的方法产生。在单一位置重复试验中评价具有单一拷贝的转基因玉米事件的麦草畏耐受性。使用来自6个构建体中的每一个的6个事件。实验设计如下:列/项目:1;处理:在V3阶段的0.5lb/a麦草畏,接着在V8阶段的1lb/a麦草畏(BASF,Raleigh,NC);重复:2;行距:30英寸;地块长度:最小20英尺;植物密度:大约30株/17.5英尺;通道(alley):2.5英尺。整个地块均匀施肥以获得农艺学上可接受的农作物。为了控制玉米根虫,在耕种时期施用每排1000英尺5盎司土壤杀虫剂,如3G(Syngenta Crop Protection,Greensboro,NC,USA)。如果观察到小地老虎(black cutworm)的侵扰,以每英亩4-8盎司的量施用3.2EC(FMC Corporation,Philadelphia,PA)。此外,使用喷洒杀虫剂计划以控制所有地上的鳞翅目害虫,包括欧洲玉米螟、玉米耳虫和秋季粘虫。每3周以每英亩4-8盎司的量施用3.2EC以控制鳞翅目害虫;大约施用4次。出土前施用如Xtra 5.6L(Monsanto,St.Louis,MO)和Degree(Monsanto,St.Louis,MO)的除草剂,以保持地决无杂草。在整个实验中,如果在未处理的检查中观察到杂草出现,则通过手工除草或者出土后施用PERMIT(Monsanto,St.Louis,MO)或者(Bayer,Research Triangle Park,NC)控制杂草。
当在V3阶段用0.5lb/a麦草畏处理接着在V8阶段用1 lb/a麦草畏处理时,通过测量支柱根损伤来测试用包含DMO转基因的DNA构建体转化的玉米近交系的麦草畏耐受性。通过计数一排中与“手指样”结构的典型形态相比显示支柱根融合的“非典型”形态的植物的数量,目视评价支柱根损伤。如表4所示,用编码未与CTP连接的DMO的DNA构建体(pMON73699、pMON73704)转化的玉米植物显示较高水平的支柱根损伤,即,麦草畏处理后的低水平保护。编码连接到CTP上的DMO的DNA构建体(pMON73716、pMON73700、pMON73715、pMON73703)显示较低水平的支柱根损伤,即,麦草畏处理后较高水平的保护。
表4.用携带DMO的DNA构建体转化的转基因玉米在测试麦草畏耐受性时显示出的支柱根损伤百分比。
近交/构建体 | 详情 | 支柱根损伤 |
01CSI6 | 对麦草畏敏感的近交系 | 95.4 |
LH244 | 对麦草畏耐受的近交系 | 93.8 |
pMON73699 | PC1SV/I-OsAct1/DMO-Wmc/TaHsp17 | 93.2 |
pMON73704 | e35S/I-OsAct1/DMO-Wmc/TaHsp17 | 91.3 |
pMON73716 | PC1SV/I-OsAct1/TaWaxy/DMO-Wmc/TaHsp17 | 78.8 |
pMON73700 | PC1SV/I-OsAct1/CTP1/DMO-Wmc/TaHsp17 | 74.4 |
pMON73715 | PC1SV/I-OsAct1/CTP2syn/DMO-Wmc/TaHsp17 | 68.2 |
pMON73703 | e35S/I-OsAct1/CTP1/DMO-Wmc/TaHsp17 | 68.8 |
从在不同的植物物种中进行的这些研究(另外,例如,实施例3、4和8)可见,叶绿体转运肽可用于DMO的高效靶向和DMO活性的完全产生,导致对麦草畏较高的耐受性。此外,在玉米中CTP-DMO的表达提供了对麦草畏的出土前耐受性。
实施例7
有效DMO表达单元的构建
一些遗传元件可以影响基因的有效表达,如启动子、叶绿体转运肽序列、内含子、5’UTR、基因的编码区、3’UTR。然而,不清楚哪些组合运行最好。需要有效的DMO表达单元或者构建体来产生改进的产物如耐受麦草畏的种子和植物。通过可操作地连接各种启动子、CTP、DMO变体和3’UTR中的各一种来构建几个DMO表达单元,以获得用于产品开发的有效的DMO表达单元。通过本领域已知的方法将这些构建体转化到大豆中(例如,U.S.6,384,301、U.S.7,002,058或Zhang等人,1999)。获得转基因种子,并检测出土前和出土后对麦草畏除草剂的耐受性。表5显示了当种子和植物在出土前用0.5lb/英亩麦草畏处理接着在出土后V6阶段用2lb/英亩的麦草畏处理时,由麦草畏造成的损伤百分比(较低的损伤意味着较高的耐受性)。用不携带CTP的DNA构建体pMON68939和pMON73723转化的种子不能耐受麦草畏的出土前施用,表明为了获得对麦草畏的出土前耐受性,需要将DMO靶向至叶绿体。用pMON68939和pMON73723(不含CTP)转化的植物,在V3后阶段用麦草畏以1lb/a的量处理后,分别显示类似于野生型大豆(60%)的55%和57%的损伤率,而用pMON68938(含有CTP)转化的植物显示很低的损伤率。这些结果表明,在大豆中获得对麦草畏的出土前和出土后耐受性需要CTP。
表5.用特异性DMO表达单元转化并在出土前和出土后用麦草畏处理的大豆植物显示的损伤百分比。
表达单元 | pMON名称 | %损伤 |
PC1SV/CTP2syn/DMO-Wat(A)/nos | 73724 | 9 |
e35S/CTP1/DMO-Wat(L)/nos | 68938 | 12 |
PC1SV/RbcSnoc/DMO-Wat(A)/nos | 73725 | 12 |
PC1SV/RbcSnoc/DMO-Wat(L)/nos | 73728 | 12 |
PCSV/CTP1/DMO-Wat(A)/nos | 73729 | 13 |
PC1SV/CTP2syn/DMO-Wat(L)/nos | 73727 | 13 |
ANT1/CTP1/DMO-Wat(L)/nos | 68945 | 14 |
PC1SV/RbcSnoc/DMO-Wat(A)/nos | 73730 | 15 |
PC1SV/RbcS-CTP/DMO-Cnat(A)/nos | 68934 | 17 |
Act7/CTP1/DMO-Wat(L)/nos | 68942 | 17 |
FMV.35S-EF1a/CTP1/DMO-Wat(L)/nos | 68940 | 17 |
PC1SV/RbcS-CTP/DMO-Cnat(A)/E9 | 84254 | 20 |
FMV/CTP1/DMO-Wat(L)/nos | 68941 | 29 |
eIF4A10/CTP1/DMO-Wat(L)/nos | 68943 | 60 |
e35S/CTP1/DMO-Cat(A)/nos | 68937 | 62 |
e35S/CTP1/DMO-Cnat(L)/nos | 68946 | 73 |
e35S/DMO-Wat(A)/nos | 68939 | 100(前) |
PC1SV/DMO-Wat(A)/nos | 73723 | 100(前) |
实施例8
耐受麦草畏的转基因棉花植物的产生
为了测试DMO基因在给棉花提供麦草畏耐受性方面的用途,产生转基因棉花植物。产生了几个DNA构建体,它们携带在植物基因表达元件如启动子(例如,PC1SV、FMV或e35S)和3’UTR(例如,E6;登记号#U30508)控制下的具有转运肽(例如,PsRbcS CTP、CTP1、CTP2)的DMO编码区(例如,SEQ ID NO:23、25、27、29、31、35),并如下所述转化到棉花(陆地棉(Gossypium hirsutum))中。所用的培养基在表6中列出。
棉花cv Coker 130的幼苗离体生长,并切下胚轴部分,并接种于携带DNA构建体的根癌土壤杆菌的液体悬浮液,印迹干燥(blot dried),并共培养2天。然后将接种的胚轴外植体转移到葡萄糖选择培养基中4周,蔗糖选择培养基中1周,葡萄糖选择培养基中再培养4周,来诱导愈伤组织。在16/8(光/暗)循环和28℃下孵育培养物。然后将卡那霉素抗性愈伤组织转移到UMO培养基中,并在28-30℃、黑暗条件下培养16-24周来诱导生胚愈伤组织。然后从这些愈伤组织中收获生胚愈伤组织,并在28-30℃、黑暗条件下、在TRP+培养基上维持长达4-16周。收获来自生胚愈伤组织的小胚,并在SHSU培养基上、在28-30℃温度下和6/8(光/暗)循环条件下发芽。然后将看起来正常的小植株转移到土壤中以获得成熟的棉花植物。转化株的转基因性质通过DNA检测来证实。
表6.用于棉花转化的各种培养基的组成。
在V4-5生长阶段以561g ae/ha(0.5lb/a)的量用麦草畏(BASF,Raleigh,NC)处理包含这样的DNA构建体的转化的棉花植物作为出土后处理,每个DNA构建体包含DMO编码区与转运肽、启动子和3’UTR的不同的组合,发现有耐受性,而未转化的棉花植物显示79%-86%的损伤率。选择显示超过95%的耐受性(等于少于5%的损伤)的转基因植物用于进一步研究。转基因植物也对随后的麦草畏出土后处理具有耐受性。例如,在V3-4阶段用0.5lb/英亩的麦草畏处理随后在V5阶段或者更晚的阶段用1或2lb/英亩的麦草畏处理的植物依然对麦草畏耐受。该实施例显示DMO基因可以向各个生长阶段的棉花提供麦草畏耐受性,因而能够在各个阶段施用麦草畏以获得有效的杂草控制。
实施例9
提高玉米直立能力的方法
如玉米的某些单子叶植物产生支柱根,支柱根从地表以上的节点生长,并有助于支撑植物和在生殖阶段从土壤上层吸收水分和营养。如果植物遇到大风或者由于根线虫感染或者地下水缺乏使得地下根***变得较弱时,健康的支柱根***变得重要。为了控制阔叶杂草,允许对如玉米的单子叶植物使用如麦草畏和2,4-D的合成除草剂。对于玉米的出土后杂草控制,麦草畏是第五广泛施用的除草剂。尽管用于控制阔叶杂草的最优量是280-560克/公顷(g/h)或者0.25-0.5lb/英亩,但是玉米中的平均使用量是168g/h或者0.15lb/英亩,因为在更高的施用量和如热天的某些环境条件下,麦草畏能够损伤玉米。此外,几种玉米杂交体,如DKC61-42、DKC64-77、DKC63-46、DKC66-21和DKC61-44,以及近交体,如01CSI6、16IUL2、70LDL5和90LCL6,对麦草畏施用敏感。敏感性以多种方式表现,如出现洋葱剥离(onion leafing)、穗畸形、株高降低或者不正常的支柱根的形成,例如融合或者扭曲的根的形成。支柱根变得多瘤节,趋向于成长在一起,并且不向土壤中生长来支撑植物。这可能导致较弱的玉米作物直立能力、较高的易倒伏性和最终的产量损失。一些含有麦草畏的除草剂产品,例如BANVEL、MARKSMAN、DISTINCT、NORTHSTAR和CELEBRITY PLUS,可以导致这些效应。提高玉米对麦草畏的耐受性在保护较接近耐受麦草畏的农作物种类(如大豆、棉花)种植的玉米地也是有用的,而且其中允许更高的麦草畏施用量。
本实施例提供了一种改进玉米和其它单子叶植物的直立能力的方法,包括在玉米中引入DMO基因并用麦草畏处理玉米。在一个实施方案中,DMO基因在组成型启动子的控制下表达,所述组成型启动子也能够在节点区域和/或支柱根中表达DMO。在另一个实施方案中,DMO基因在一种嵌合组成型和节点/支柱根特异性启动子的控制下表达。在另一个实施方案中,DMO基因在根特异性启动子如RCc3或者其变体(例如,在US20060101541中发现的SEQ ID NOs:1-6)的控制下表达。DMO在支柱根中的表达导致支柱根没有或者有较少的损伤,导致玉米的较高的直立能力、较少的倒伏,因而导致较高的产量。
来自用各种DMO构建体(在表7中列出)转化的玉米植物的3个单拷贝事件的R1或者F1种子在4.0”托盘中发芽。将健康植物移植到大约10”的罐中。发芽和生长培养基包括Redi-earthTM (Scotts-SierraHorticultural Products Co.,Marysville,Ohio)。将罐放在35英寸×60英寸的玻璃纤维浇水托盘中的毛细管垫上,用于在测试期间进行亚浇灌,从而保持对于植物生长而言最佳的土壤含水量。用Osmocote (14-14-14缓释;Scotts-Sierra Horticultural Products Co.,Marysville,Ohio)以100gm/cu.ft.的量对罐施肥,以在温室试验期间维持植物生长。植物在29℃/21℃的昼/夜温度、25%-75%的相对湿度的温室中生长,以模拟晚春温暖季节的生长条件。按照需要,为最少14小时的光照周期提供大约600μE的补充光。
使用具有设定为最小24psi(165kpa)的大气压的Teejet 9501E平扇喷嘴(Spraying Systems Co,Wheaton,IL),用轨道喷雾器来施用麦草畏。喷嘴保持在比植物顶端高大约16英寸的位置进行喷雾。喷雾体积是10加仑/英亩或者93升/公顷。当植物达到V4-5叶阶段时,开始施用。
通过在V4-5阶段用21b/英亩或者4lb/英亩的麦草畏处理,并在处理后24天评估植物的支柱根损伤(0%;没有可见的损伤,到100%,植物完全死亡)和倒伏(倾斜程度),来检测用包含DMO表达单元的DNA构建体转化的玉米近交系的植物的支柱根损伤和倒伏情况。
如表7所示,与未被转化的对照近交系和只用选择性标记表达单元(pMON73746)转化的植物相比较,用含有DMO表达单元的DNA构建体转化的玉米植物显示没有或者几乎没有支柱根损伤和倒伏。本实施例显示当用麦草畏处理时,包含DMO的植物可以被用来提供提高的直立能力。
表7.当用麦草畏处理时,用各种DMO构建体转化的玉米植物显示没有或者几乎没有支柱根损伤和倒伏。
参考文献
以下列出的文献引入本文作为参考,以补充、解释、提供背景或教导本文采用的方法、技术和/或组合物。
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Claims (9)
1.一种重组DNA分子,其包括可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码SEQ ID NO:6的多肽序列。
2.如权利要求1所述的重组DNA分子,其中,所述编码叶绿体转运肽的DNA序列如SEQ ID NO:17的核苷酸序列所示。
3.如权利要求1所述的重组DNA分子,其中,所述编码麦草畏单加氧酶的DNA序列编码选自SEQ ID NO:24、26、28、30、32、34、36、38和40的多肽。
4.如权利要求3所述的重组DNA分子,其中,所述编码麦草畏单加氧酶的DNA序列选自SEQ ID NO:23、25、27、29、31、33、35、37和39。
5.如权利要求1所述的重组DNA分子,进一步包含启动子。
6.一种产生耐受麦草畏的植物的方法,包括:将包含可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列的重组DNA分子引入植物细胞中,其中,编码叶绿体转运肽的DNA序列编码SEQ ID NO:6的序列;并由该植物细胞再生包含该重组DNA分子的植物。
7.如权利要求6所述的方法,其中,所述植物是双子叶植物。
8.如权利要求6所述的方法,其中,所述植物是单子叶植物。
9.一种控制田地中的杂草的方法,包括:培育包含重组DNA分子的转基因植物,该重组DNA分子包含可操作地连接到编码麦草畏单加氧酶的DNA序列上的编码叶绿体转运肽的DNA序列,其中,编码叶绿体转运肽的DNA序列编码SEQ ID NO:6的序列;并向所述田地中施用有效控制杂草生长而不损伤所述转基因植物的量的麦草畏除草剂。
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US9901057B2 (en) | 2015-06-29 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01050566 |
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US9907271B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01056960 |
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US9832965B2 (en) | 2015-06-30 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01051604 |
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TW201718862A (zh) | 2015-09-22 | 2017-06-01 | Dow Agrosciences Llc | 用於轉殖基因表現之植物啟動子及3’utr |
TW201718861A (zh) | 2015-09-22 | 2017-06-01 | 道禮責任有限公司 | 用於轉殖基因表現之植物啟動子及3’utr |
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US9894862B2 (en) | 2015-09-24 | 2018-02-20 | Monsanto Technology Llc | Soybean variety 01058666 |
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US9907274B2 (en) | 2015-09-24 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01052030 |
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WO2017070298A1 (en) | 2015-10-22 | 2017-04-27 | Dow Agrosciences Llc | Plant promoter for transgene expression |
US20170121726A1 (en) | 2015-11-04 | 2017-05-04 | Dow Agrosciences Llc | Plant promoter for transgene expression |
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US9854762B2 (en) | 2015-12-18 | 2018-01-02 | Monsanto Technology Llc | Cotton variety 15R509B2XF |
US9854761B2 (en) | 2015-12-18 | 2018-01-02 | Monsanto Technology Llc | Cotton variety 15R510B2XF |
US9961845B2 (en) | 2015-12-18 | 2018-05-08 | Monsanto Technology Llc | Cotton variety 15R551B2XF |
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US9585355B1 (en) | 2016-03-21 | 2017-03-07 | Monsanto Technology Llc | Soybean variety XR40AA15X |
US9585354B1 (en) | 2016-03-21 | 2017-03-07 | Monsanto Technology Llc | Soybean variety XR48C15X |
US9763413B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR23AQ15X |
US9560827B1 (en) | 2016-03-21 | 2017-02-07 | Monsanto Technology Llc | Soybean variety XR45C15X |
US9603331B1 (en) | 2016-03-21 | 2017-03-28 | Monsanto Technology Llc | Soybean variety XR39V15X |
US9596819B1 (en) | 2016-03-21 | 2017-03-21 | Monsanto Technology Llc | Soybean variety XR33AU15X |
US9861053B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR36AC15X |
US9788508B1 (en) | 2016-03-21 | 2017-10-17 | Monsanto Technology Llc | Soybean variety XR20AH15X |
US9861054B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR36Z15X |
US9565822B1 (en) | 2016-03-21 | 2017-02-14 | Monsanto Technology Llc | Soybean variety XR37AE15X |
US9763414B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR25AL15X |
US9861055B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR39U15X |
US9763412B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR09AP15X |
US9730415B1 (en) | 2016-03-21 | 2017-08-15 | Monsanto Technology Llc | Soybean variety XR08A15X |
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US9706739B1 (en) | 2016-06-01 | 2017-07-18 | Monsanto Technology Llc | Soybean variety 01057290 |
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US9832953B1 (en) | 2016-06-01 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01057348 |
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US9907250B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057493 |
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US9907249B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057459 |
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US9788514B1 (en) | 2016-06-01 | 2017-10-17 | Monsanto Technology Llc | Soybean variety 01057345 |
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US10226002B2 (en) | 2016-06-01 | 2019-03-12 | Monsanto Technology Llc | Soybean variety 01057408 |
US9848563B1 (en) | 2016-06-01 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01058353 |
US9907251B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058313 |
US9968054B2 (en) | 2016-06-01 | 2018-05-15 | Monsanto Technology Llc | Soybean variety 01057207 |
US9907252B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058320 |
US9693530B1 (en) | 2016-06-02 | 2017-07-04 | Monsanto Technology Llc | Soybean variety 01063884 |
US9832955B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01058718 |
US9700009B1 (en) | 2016-06-02 | 2017-07-11 | Monsanto Technology Llc | Soybean variety 01058861 |
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US9867353B2 (en) | 2016-06-02 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01058528 |
US9832954B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01058711 |
US9907255B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058767 |
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US9901053B2 (en) | 2016-06-02 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01058606 |
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US9907253B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058513 |
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US9743626B1 (en) | 2016-06-06 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01064148 |
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US9844197B1 (en) | 2016-06-06 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01064151 |
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US9867355B2 (en) | 2016-06-06 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01064176 |
US9844198B1 (en) | 2016-06-07 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01063894 |
US9848568B1 (en) | 2016-06-07 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01064130 |
US9743629B1 (en) | 2016-06-07 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01058714 |
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EP3519574B1 (en) | 2016-10-03 | 2022-01-19 | Corteva Agriscience LLC | Plant promoter for transgene expression |
BR112019005687A2 (pt) | 2016-10-03 | 2019-07-02 | Dow Agrosciences Llc | promotor vegetal para expressão de transgenes |
WO2018085693A1 (en) | 2016-11-04 | 2018-05-11 | Inari Agriculture, Inc. | Novel plant cells, plants, and seeds |
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CN110475866A (zh) | 2017-01-30 | 2019-11-19 | 科沃施种子欧洲股份两合公司 | 用于基因组工程的与核酸内切酶相连的修复模板 |
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US9888659B1 (en) | 2017-03-29 | 2018-02-13 | Monsanto Technology Llc | Soybean cultivar 61040705 |
US10039249B1 (en) | 2017-03-29 | 2018-08-07 | Monsanto Technology Llc | Soybean cultivar 61162227 |
US9930856B1 (en) | 2017-03-29 | 2018-04-03 | Monsanto Technology Llc | Soybean cultivar 62030513 |
US9961868B1 (en) | 2017-03-29 | 2018-05-08 | Monsanto Technology Llc | Soybean cultivar 60111034 |
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US9913449B1 (en) | 2017-03-29 | 2018-03-13 | Monsanto Technology Llc | Soybean cultivar 67272133 |
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US9907281B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 64291310 |
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US9907280B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 64492916 |
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US9907282B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 60371632 |
US10368518B2 (en) | 2017-06-26 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01068117 |
US10039254B1 (en) | 2017-06-26 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064117 |
US10314273B2 (en) | 2017-06-26 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01068110 |
US10405511B2 (en) | 2017-06-26 | 2019-09-10 | Monsanto Technology Llc | Soybean variety 01064350 |
US10383304B2 (en) | 2017-06-26 | 2019-08-20 | Monsanto Technology Llc | Soybean variety 01063944 |
US10034448B1 (en) | 2017-06-26 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064516 |
US10342197B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01064349 |
US10342199B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01068883 |
US10010046B1 (en) | 2017-06-26 | 2018-07-03 | Monsanto Technology Llc | Soybean variety 01064401 |
US10314274B2 (en) | 2017-06-26 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01068109 |
US10455788B2 (en) | 2017-06-26 | 2019-10-29 | Monsanto Technology Llc | Soybean variety 01068888 |
US10034447B1 (en) | 2017-06-26 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064421 |
US10342198B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01064381 |
US10039255B1 (en) | 2017-06-26 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064252 |
US10194613B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064363 |
US10244704B2 (en) | 2017-06-28 | 2019-04-02 | Monsanto Technologies Llc | Soybean variety 01068084 |
US10196646B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064344 |
US10206350B1 (en) | 2017-06-28 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 5PDSD70 |
US10178837B1 (en) | 2017-06-28 | 2019-01-15 | Monsanto Technology Llc | Soybean variety 5PHQP31 |
US10053700B1 (en) | 2017-06-28 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064288 |
US10368511B2 (en) | 2017-06-28 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01068085 |
US10172310B1 (en) | 2017-06-28 | 2019-01-08 | Monsanto Technology Llc | Soybean variety 5PYFS21 |
US10244705B2 (en) | 2017-06-28 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064265 |
US10011845B1 (en) | 2017-06-28 | 2018-07-03 | Monsanto Technology Llc | Soybean variety 01068893 |
US10206349B1 (en) | 2017-06-28 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 5PPHU37 |
US10368512B2 (en) | 2017-06-28 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01068083 |
US10178838B1 (en) | 2017-06-28 | 2019-01-15 | Monsanto Technology Llc | Soybean variety 5PQNG18 |
US10188059B1 (en) | 2017-06-28 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 5PUWT62 |
US10053699B1 (en) | 2017-06-28 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01068892 |
US10292358B2 (en) | 2017-06-28 | 2019-05-21 | Monsanto Technology Llc | Soybean variety 01064327 |
US10194612B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01068093 |
US10182539B1 (en) | 2017-06-28 | 2019-01-22 | Monsanto Technology Llc | Soybean variety 5PDZB74 |
US10201138B2 (en) | 2017-06-28 | 2019-02-12 | Monsanto Technology Llc | Soybean variety 01064519 |
US10271493B2 (en) | 2017-06-28 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064280 |
US10036027B1 (en) | 2017-06-28 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064536 |
US10349598B2 (en) | 2017-06-28 | 2019-07-16 | Monsanto Technology Llc | Soybean variety 01068088 |
US10194614B2 (en) | 2017-07-06 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064567 |
US10201139B2 (en) | 2017-07-06 | 2019-02-12 | Monsanto Technology Llc | Soybean variety 01068144 |
US10244706B2 (en) | 2017-07-06 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064571 |
US10064379B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064450 |
US10238057B2 (en) | 2017-07-06 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064461 |
US10070617B1 (en) | 2017-07-06 | 2018-09-11 | Monsanto Technology Llc | Soybean variety 01063975 |
US10064382B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064035 |
US10098305B1 (en) | 2017-07-06 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01064403 |
US10064381B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01067510 |
US10231403B2 (en) | 2017-07-06 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068141 |
US10070616B1 (en) | 2017-07-06 | 2018-09-11 | Monsanto Technology Llc | Soybean variety 01067507 |
US10064367B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01067515 |
US10194616B2 (en) | 2017-07-06 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064568 |
US10238076B2 (en) | 2017-07-19 | 2019-03-26 | Monsanto Technology, Llc | Soybean variety 01068130 |
US10212901B2 (en) | 2017-07-19 | 2019-02-26 | Monsanto Technology Llc | Soybean variety 01068895 |
US10231404B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064187 |
US10398105B2 (en) | 2017-07-19 | 2019-09-03 | Monsanto Technology Llc | Soybean variety 01064198 |
US10058061B1 (en) | 2017-07-19 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01068112 |
US10064368B1 (en) | 2017-07-19 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01068133 |
US10098306B1 (en) | 2017-07-19 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01068070 |
US10231422B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068134 |
US10136602B1 (en) | 2017-07-19 | 2018-11-27 | Monsanto Technology Llc | Soybean variety 01068087 |
US10356997B2 (en) | 2017-07-19 | 2019-07-23 | Monsanto Technology Llc | Soybean variety 01068891 |
US10206351B2 (en) | 2017-07-19 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 01068060 |
US10194617B1 (en) | 2017-07-19 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01068066 |
US10219466B2 (en) | 2017-07-19 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064243 |
US10231421B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064231 |
US10231405B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068135 |
US10314275B2 (en) | 2017-07-19 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064238 |
US10058060B1 (en) | 2017-07-19 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01068082 |
US10426116B2 (en) | 2017-07-19 | 2019-10-01 | Monsanto Technology Llc | Soybean variety 01068124 |
US10206352B2 (en) | 2017-07-19 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 01068065 |
US10440908B2 (en) | 2017-07-19 | 2019-10-15 | Monsanto Technology Llc | Soybean variety 01064245 |
US10064369B1 (en) | 2017-07-19 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01068131 |
US10206353B2 (en) | 2017-07-20 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 01064485 |
US10244723B2 (en) | 2017-07-20 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01068058 |
US10098307B1 (en) | 2017-07-20 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01064838 |
US10064370B1 (en) | 2017-07-20 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064806 |
US10117401B1 (en) | 2017-07-20 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064488 |
US10143167B1 (en) | 2017-07-20 | 2018-12-04 | Monsanto Technology Llc | Soybean variety 01064871 |
US10285367B2 (en) | 2017-07-20 | 2019-05-14 | Monsanto Technology Llc | Soybean variety 01064508 |
US10231406B2 (en) | 2017-07-20 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068831 |
US10098308B1 (en) | 2017-07-20 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01068054 |
US10117400B1 (en) | 2017-07-20 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064185 |
US10051823B1 (en) | 2017-07-20 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064868 |
US10058062B1 (en) | 2017-07-20 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01064840 |
US10368513B2 (en) | 2017-07-31 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01064206 |
US10117402B1 (en) | 2017-07-31 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01068884 |
US10231407B1 (en) | 2017-08-23 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01063959 |
US10117404B1 (en) | 2017-08-23 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064613 |
US10271494B2 (en) | 2017-08-23 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01068122 |
US10231408B1 (en) | 2017-08-23 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064013 |
US10257994B2 (en) | 2017-08-23 | 2019-04-16 | Monsanto Technology Llc | Soybean variety 01068143 |
US10264748B2 (en) | 2017-08-23 | 2019-04-23 | Monsanto Technology Llc | Soybean variety 01064212 |
US10244708B2 (en) | 2017-08-23 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064594 |
US10117403B1 (en) | 2017-08-23 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064099 |
US10285368B2 (en) | 2017-08-23 | 2019-05-14 | Monsanto Technology Llc | Soybean variety 01064056 |
US10238059B2 (en) | 2017-08-23 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064460 |
US10251354B2 (en) | 2017-08-23 | 2019-04-09 | Monsanto Technology Llc | Soybean variety 01064596 |
US10136603B1 (en) | 2017-08-24 | 2018-11-27 | Monsanto Technology Llc | Soybean variety 01067535 |
US10045507B1 (en) | 2017-08-24 | 2018-08-14 | Monsanto Technology Llc | Soybean variety 01064406 |
US10123501B1 (en) | 2017-08-24 | 2018-11-13 | Monsanto Technology Llc | Soybean variety 01064722 |
US10039256B1 (en) | 2017-08-24 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064446 |
US10130059B1 (en) | 2017-08-24 | 2018-11-20 | Monsanto Technology Llc | Soybean variety 01068801 |
US10342200B2 (en) | 2017-08-24 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01067525 |
US10051825B1 (en) | 2017-08-24 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064400 |
US10314276B2 (en) | 2017-08-24 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064370 |
US10045506B1 (en) | 2017-08-24 | 2018-08-14 | Monsanto Technology Llc | Soybean variety 01064415 |
US10051824B1 (en) | 2017-08-24 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064393 |
US10130060B1 (en) | 2017-08-24 | 2018-11-20 | Monsanto Technology Llc | Soybean variety 01068804 |
US10130058B1 (en) | 2017-08-24 | 2018-11-20 | Monsanto Technology Llc | Soybean variety 01064723 |
US10231423B1 (en) | 2017-08-24 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064404 |
US10231409B1 (en) | 2017-08-29 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064026 |
US10485197B2 (en) | 2017-08-29 | 2019-11-26 | Monsanto Technology Llc | Soybean variety 01064625 |
US10271497B2 (en) | 2017-08-29 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064054 |
US10159205B1 (en) | 2017-08-29 | 2018-12-25 | Monsanto Technology Llc | Soybean variety 01064633 |
US10244724B2 (en) | 2017-08-29 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064667 |
US10271496B2 (en) | 2017-08-29 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064102 |
US10257995B2 (en) | 2017-08-29 | 2019-04-16 | Monsanto Technology Llc | Soybean variety 01064083 |
US10271495B2 (en) | 2017-08-29 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064036 |
US10238077B2 (en) | 2017-08-29 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064104 |
US10314265B2 (en) | 2017-08-29 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064669 |
US10342201B2 (en) | 2017-08-29 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01067531 |
US10314277B2 (en) | 2017-08-29 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064636 |
US10433504B2 (en) | 2017-08-29 | 2019-10-08 | Monsanto Technology Llc | Soybean variety 01064105 |
US10159206B1 (en) | 2017-08-29 | 2018-12-25 | Monsanto Technology Llc | Soybean variety 01067519 |
US10231426B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064748 |
US10271498B2 (en) | 2017-08-30 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064676 |
US10188072B1 (en) | 2017-08-30 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 01064694 |
US10117405B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064511 |
US10051826B1 (en) | 2017-08-30 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064189 |
US10117409B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064756 |
US10231424B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01067526 |
US10231425B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01067534 |
US10188061B1 (en) | 2017-08-30 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 01064486 |
US10194618B1 (en) | 2017-08-30 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064184 |
US10219467B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064677 |
US10117407B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064490 |
US10342202B2 (en) | 2017-08-30 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01064775 |
US10064371B1 (en) | 2017-08-30 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064809 |
US10219484B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064735 |
US10219485B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01069661 |
US10051827B1 (en) | 2017-08-30 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064164 |
US10117406B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064166 |
US10117408B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064491 |
US10212902B1 (en) | 2017-08-30 | 2019-02-26 | Monsanto Technology Llc | Soybean variety 01067523 |
US10314278B2 (en) | 2017-08-30 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064635 |
US10278350B2 (en) | 2017-08-30 | 2019-05-07 | Monsanto Technology Llc | Soybean variety 01064672 |
US10357007B2 (en) | 2017-08-30 | 2019-07-23 | Monsanto Technology Llc | Soybean variety 01064743 |
US10863715B2 (en) | 2017-12-28 | 2020-12-15 | Monsanto Technology Llc | Cotton variety 16R343B3XF |
US10842121B2 (en) | 2017-12-28 | 2020-11-24 | Monsanto Technology Llc | Cotton variety 16R141XF |
US10842120B2 (en) | 2017-12-28 | 2020-11-24 | Monsanto Technology Llc | Cotton variety 16R125XF |
US10856513B2 (en) | 2017-12-28 | 2020-12-08 | Monsanto Technology Llc | Cotton variety 16R338B3XF |
US10863713B2 (en) | 2017-12-28 | 2020-12-15 | Monsanto Technology Llc | Cotton variety 16R341B3XF |
US10842122B2 (en) | 2017-12-28 | 2020-11-24 | Monsanto Technology Llc | Cotton variety 16R123XF |
US10863714B2 (en) | 2017-12-28 | 2020-12-15 | Monsanto Technology Llc | Cotton variety 16R330B3XF |
US10939656B2 (en) | 2017-12-28 | 2021-03-09 | Monsanto Technology Llc | Cotton variety 16R228NRB2XF |
US10849302B2 (en) | 2017-12-28 | 2020-12-01 | Monsanto Technology Llc | Cotton variety 16R335B3XF |
US10925248B2 (en) | 2017-12-28 | 2021-02-23 | Monsanto Technology Llc | Cotton variety 16R225NRB2XF |
US10869455B2 (en) | 2017-12-28 | 2020-12-22 | Monsanto Technology Llc | Cotton variety 16R345B3XF |
US10856512B2 (en) | 2017-12-28 | 2020-12-08 | Monsanto Technology Llc | Cotton variety 16R336B3XF |
US10842123B2 (en) | 2017-12-28 | 2020-11-24 | Monsanto Technology Llc | Cotton variety 16R324B3XF |
US10874079B2 (en) | 2017-12-28 | 2020-12-29 | Monsanto Technology Llc | Cotton variety 16R351B3XF |
US10368519B1 (en) | 2018-05-24 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 5PYGS18 |
US10368520B1 (en) | 2018-05-24 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 5PCDJ10 |
US10420312B1 (en) | 2018-05-24 | 2019-09-24 | Monsanto Technology Llc | Soybean variety 5PKGL32 |
US10412921B1 (en) | 2018-05-24 | 2019-09-17 | Monsanto Technology Llc | Soybean variety 5PYUP46 |
US10477818B1 (en) | 2018-05-24 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 5PTJB40 |
US10420313B1 (en) | 2018-05-24 | 2019-09-24 | Monsanto Technology Llc | Soybean variety 5PKVF24 |
US10480005B1 (en) | 2018-05-24 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 5PEKB42 |
US10258013B1 (en) | 2018-08-06 | 2019-04-16 | Monsanto Technology Llc | Soybean cultivar 64442216 |
US10398122B1 (en) | 2018-08-06 | 2019-09-03 | Monsanto Technology Llc | Soybean cultivar 77252418 |
US10264760B1 (en) | 2018-08-06 | 2019-04-23 | Monsanto Technology Llc | Soybean cultivar 79380729 |
US10342207B1 (en) | 2018-08-06 | 2019-07-09 | Monsanto Technology Llc | Soybean cultivar 68381333 |
US10368521B1 (en) | 2018-08-06 | 2019-08-06 | Monsanto Technology Llc | Soybean cultivar 70490447 |
US10321654B1 (en) | 2018-08-06 | 2019-06-18 | Monsanto Technology Llc | Soybean cultivar 76222418 |
US10238080B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 79072143 |
US10238066B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 73301323 |
US10517255B1 (en) | 2018-08-06 | 2019-12-31 | Monsanto Technology Llc | Soybean cultivar 62391708 |
US10258014B1 (en) | 2018-08-06 | 2019-04-16 | Monsanto Technology Llc | Soybean cultivar 76051047 |
US10238079B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 77301427 |
US10362758B1 (en) | 2018-08-06 | 2019-07-30 | Monsanto Technology Llc | Soybean cultivar 64272401 |
US10342206B1 (en) | 2018-08-06 | 2019-07-09 | Monsanto Technology Llc | Soybean cultivar 60390131 |
US10349608B1 (en) | 2018-08-06 | 2019-07-16 | Monsanto Technology Llc | Soybean cultivar 70520427 |
US10357010B1 (en) | 2018-08-06 | 2019-07-23 | Monsanto Technology Llc | Soybean cultivar 68352516 |
US10342204B1 (en) | 2018-08-06 | 2019-07-09 | Monsanto Technology Llc | Soybean cultivar 65382631 |
US10349607B1 (en) | 2018-08-06 | 2019-07-16 | Monsanto Technology Llc | Soybean cultivar 78091047 |
US10624285B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01068376 |
US10617089B2 (en) | 2018-08-22 | 2020-04-14 | Monsanto Technology Llc | Soybean variety 01067827 |
US10624303B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072279 |
US10555482B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068427 |
US11013197B2 (en) | 2018-08-22 | 2021-05-25 | Monsanto Technology Llc | Soybean variety 01072368 |
US10555484B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068903 |
US10973200B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072342 |
US10561098B1 (en) | 2018-08-22 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01067649 |
US10999998B2 (en) | 2018-08-22 | 2021-05-11 | Monsanto Technology Llc | Soybean variety 01072346 |
US10993404B2 (en) | 2018-08-22 | 2021-05-04 | Monsanto Technology Llc | Soybean variety 01072356 |
US11206791B2 (en) | 2018-08-22 | 2021-12-28 | Monsanto Technology Llc | Soybean variety 01072337 |
US10624302B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072280 |
US10905080B2 (en) | 2018-08-22 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072355 |
US10918065B2 (en) | 2018-08-22 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072343 |
US11096363B2 (en) | 2018-08-22 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01072331 |
US10561099B1 (en) | 2018-08-22 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068352 |
US10897868B2 (en) | 2018-08-22 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01072327 |
US10582686B1 (en) | 2018-08-22 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01064580 |
US10555481B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068364 |
US11109552B2 (en) | 2018-08-22 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01072332 |
US10897870B2 (en) | 2018-08-22 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01072322 |
US10939652B2 (en) | 2018-08-22 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072350 |
US10575489B1 (en) | 2018-08-22 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01067545 |
US10973201B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072339 |
US10575490B1 (en) | 2018-08-22 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01067547 |
US10602689B2 (en) | 2018-08-22 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068908 |
US10602704B2 (en) | 2018-08-22 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068313 |
US11109553B2 (en) | 2018-08-22 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01072328 |
US10555483B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067598 |
US10973198B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072340 |
US11026389B2 (en) | 2018-08-22 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01072367 |
US10905081B2 (en) | 2018-08-22 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072329 |
US11044876B2 (en) | 2018-08-22 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01072345 |
US10897869B2 (en) | 2018-08-22 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01072333 |
US10973199B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072341 |
US10555480B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068350 |
US10524446B1 (en) | 2018-08-23 | 2020-01-07 | Monsanto Technology Llc | Soybean variety 01072245 |
US10609888B2 (en) | 2018-08-23 | 2020-04-07 | Monsanto Technology Llc | Soybean variety 01068346 |
US10624287B2 (en) | 2018-08-23 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072275 |
US10631486B2 (en) | 2018-08-23 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068187 |
US10624286B2 (en) | 2018-08-23 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072276 |
US10448608B1 (en) | 2018-08-23 | 2019-10-22 | Monsanto Technology Llc | Soybean variety 01067812 |
US10448609B1 (en) | 2018-08-23 | 2019-10-22 | Monsanto Technology Llc | Soybean variety 01072293 |
US10512239B1 (en) | 2018-08-23 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072273 |
US10506786B1 (en) | 2018-08-23 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01067789 |
US10492450B1 (en) | 2018-08-23 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01064752 |
US10463000B1 (en) | 2018-08-23 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01072224 |
US10499601B1 (en) | 2018-08-23 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01068356 |
US10638708B2 (en) | 2018-08-23 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068325 |
US10687501B2 (en) | 2018-08-23 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068438 |
US10548284B1 (en) | 2018-08-23 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01072294 |
US10555485B1 (en) | 2018-08-23 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01073532 |
US10638707B2 (en) | 2018-08-23 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01067639 |
US10687502B2 (en) | 2018-08-23 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068433 |
US10631499B2 (en) | 2018-08-23 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01072291 |
US10555486B1 (en) | 2018-08-23 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068910 |
US10631503B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067814 |
US10470425B1 (en) | 2018-08-24 | 2019-11-12 | Monsanto Technology Llc | Soybean variety 01067783 |
US10531629B1 (en) | 2018-08-24 | 2020-01-14 | Monsanto Technology Llc | Soybean variety 01072223 |
US10512240B1 (en) | 2018-08-24 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072228 |
US10631500B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067788 |
US10561106B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068466 |
US10463010B1 (en) | 2018-08-24 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01068189 |
US10631501B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067793 |
US10638710B2 (en) | 2018-08-24 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068195 |
US10561102B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068182 |
US10631504B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01072295 |
US10561101B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068184 |
US10631505B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01069676 |
US10485213B1 (en) | 2018-08-24 | 2019-11-26 | Monsanto Technology Llc | Soybean variety 01068968 |
US10582687B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01068359 |
US10631502B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067840 |
US10602705B2 (en) | 2018-08-24 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068821 |
US10548285B1 (en) | 2018-08-24 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01067703 |
US10631507B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068485 |
US10617090B2 (en) | 2018-08-24 | 2020-04-14 | Monsanto Technology Llc | Soybean variety 01068163 |
US10470395B1 (en) | 2018-08-24 | 2019-11-12 | Monsanto Technology Llc | Soybean variety 01067663 |
US10582690B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067790 |
US10624304B2 (en) | 2018-08-24 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01068381 |
US10555488B1 (en) | 2018-08-24 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067714 |
US10561100B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01067742 |
US10660292B2 (en) | 2018-08-24 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01068157 |
US10645898B2 (en) | 2018-08-24 | 2020-05-12 | Monsanto Technology Llc | Soybean variety 01068192 |
US10575491B1 (en) | 2018-08-24 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01072289 |
US10561104B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01072234 |
US10561105B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01072235 |
US10582688B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067543 |
US10582689B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067573 |
US10492451B1 (en) | 2018-08-24 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 0107226 |
US10506787B1 (en) | 2018-08-24 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01072220 |
US10631506B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068838 |
US10561103B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01073501 |
US10542721B1 (en) | 2018-08-24 | 2020-01-28 | Monsanto Technology Llc | Soybean variety 01072231 |
US10638709B2 (en) | 2018-08-24 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068191 |
US10448610B1 (en) | 2018-08-24 | 2019-10-22 | Monsanto Technology Llc | Soybean variety 01067781 |
US10555487B1 (en) | 2018-08-24 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067694 |
US10517259B1 (en) | 2018-08-24 | 2019-12-31 | Monsanto Technology Llc | Soybean variety 01068824 |
US10477821B1 (en) | 2018-08-24 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01067769 |
US10512241B1 (en) | 2018-08-24 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072232 |
US10463012B1 (en) | 2018-08-27 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01069688 |
US10575492B1 (en) | 2018-08-27 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01068915 |
US10477823B1 (en) | 2018-08-27 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01069720 |
US10602690B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068227 |
US10492452B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01067646 |
US10743492B2 (en) | 2018-08-27 | 2020-08-18 | Monsanto Technology Llc | Soybean variety 01077433 |
US10575493B1 (en) | 2018-08-27 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01067819 |
US10743491B2 (en) | 2018-08-27 | 2020-08-18 | Monsanto Technology Llc | Soybean variety 01077432 |
US10463011B1 (en) | 2018-08-27 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01068162 |
US10492454B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01072256 |
US10477822B1 (en) | 2018-08-27 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01069686 |
US10602706B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01077437 |
US10492453B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01072254 |
US10582678B1 (en) | 2018-08-27 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01072239 |
US10499603B1 (en) | 2018-08-27 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01068207 |
US10561107B1 (en) | 2018-08-27 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068807 |
US10743493B2 (en) | 2018-08-27 | 2020-08-18 | Monsanto Technology Llc | Soybean variety 01077438 |
US10512242B1 (en) | 2018-08-27 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01077436 |
US10506788B1 (en) | 2018-08-27 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01072292 |
US10568291B1 (en) | 2018-08-27 | 2020-02-25 | Monsanto Technology Llc | Soybean variety 01067741 |
US10660293B2 (en) | 2018-08-27 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01077434 |
US10499602B1 (en) | 2018-08-27 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01067553 |
US10568290B1 (en) | 2018-08-27 | 2020-02-25 | Monsanto Technology Llc | Soybean variety 01068219 |
US10548286B1 (en) | 2018-08-27 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01068171 |
US10660294B2 (en) | 2018-08-27 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01077435 |
US10602692B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01072252 |
US10631508B2 (en) | 2018-08-27 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068228 |
US10602691B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01072253 |
US10555489B1 (en) | 2018-08-27 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068972 |
US10582679B1 (en) | 2018-08-27 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01072243 |
US10463013B1 (en) | 2018-08-27 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01069709 |
US10687503B2 (en) | 2018-08-27 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068213 |
US10631509B2 (en) | 2018-08-27 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067807 |
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US10506789B1 (en) | 2018-12-04 | 2019-12-17 | Monsanto Technology Llc | Soybean cultivar 61381330 |
US10918070B2 (en) | 2018-12-19 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R806B3XF |
US10918071B2 (en) | 2018-12-19 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 16R346B3XF |
US10905086B2 (en) | 2018-12-19 | 2021-02-02 | Monsanto Technology Llc | Cotton variety 17R820B3XF |
US10905085B2 (en) | 2018-12-19 | 2021-02-02 | Monsanto Technology Llc | Cotton variety 17R808B3XF |
US10874081B2 (en) | 2018-12-19 | 2020-12-29 | Monsanto Technology Llc | Cotton variety 17R709XF |
US10918074B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R844B3XF |
US10905087B2 (en) | 2019-01-09 | 2021-02-02 | Monsanto Technology Llc | Cotton variety 16R246NRB2XF |
US10925250B2 (en) | 2019-01-09 | 2021-02-23 | Monsanto Technology Llc | Cotton variety 17R816B3XF |
US10925249B2 (en) | 2019-01-09 | 2021-02-23 | Monsanto Technology Llc | Cotton variety 17R933NRB3XF |
US10939661B2 (en) | 2019-01-09 | 2021-03-09 | Monsanto Technology Llc | Cotton variety 17R815B3XF |
US10939658B2 (en) | 2019-01-09 | 2021-03-09 | Monsanto Technology Llc | Cotton variety 16R232B2XF |
US10918075B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R845B3XF |
US10939660B2 (en) | 2019-01-09 | 2021-03-09 | Monsanto Technology Llc | Cotton variety 17R740XF |
US10939659B2 (en) | 2019-01-09 | 2021-03-09 | Monsanto Technology Llc | Cotton variety 16R353B3XF |
US10918034B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R737XF |
US10918076B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R817B3XF |
US10918072B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R814B3XF |
US10918077B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R827B3XF |
US10918073B2 (en) | 2019-01-09 | 2021-02-16 | Monsanto Technology Llc | Cotton variety 17R819B3XF |
EP3935156A4 (en) | 2019-03-07 | 2022-12-28 | The Regents of The University of California | CRISPR-CAS EFFECTOR POLYPEPTIDES AND METHODS OF USE |
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US10617091B1 (en) | 2019-06-04 | 2020-04-14 | Monsanto Technology Llc | Soybean cultivar 86331141 |
US10624305B1 (en) | 2019-06-04 | 2020-04-21 | Monsanto Technology Llc | Soybean cultivar 85161422 |
US10602693B1 (en) | 2019-06-04 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 81302732 |
US10631513B1 (en) | 2019-06-04 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 87181312 |
US10667484B1 (en) | 2019-06-04 | 2020-06-02 | Monsanto Technology Llc | Soybean cultivar 82322805 |
US10631512B1 (en) | 2019-06-04 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 85322536 |
US10548287B1 (en) | 2019-06-05 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 86411138 |
US10638713B1 (en) | 2019-06-05 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 85220103 |
US10638686B1 (en) | 2019-06-05 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 84161023 |
US10624288B1 (en) | 2019-06-05 | 2020-04-21 | Monsanto Technology Llc | Soybean cultivar 85061635 |
US10660295B1 (en) | 2019-06-05 | 2020-05-26 | Monsanto Technology Llc | Soybean cultivar 86072029 |
US10609883B1 (en) | 2019-06-05 | 2020-04-07 | Monsanto Technology Llc | Soybean cultivar 82340103 |
US10602707B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 85442619 |
US10602709B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 86360847 |
US10631514B1 (en) | 2019-06-05 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 80131402 |
US10602708B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 82370721 |
US10617092B1 (en) | 2019-06-06 | 2020-04-14 | Monsanto Technology Llc | Soybean cultivar 80411617 |
US10561108B1 (en) | 2019-06-06 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 80431932 |
US10568292B1 (en) | 2019-06-06 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 80181322 |
US10548288B1 (en) | 2019-06-06 | 2020-02-04 | Mansanto Technology Llc | Soybean cultivar 80372800 |
US10548273B1 (en) | 2019-06-06 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 87120510 |
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US10631487B1 (en) | 2019-06-06 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 83240828 |
US10548289B1 (en) | 2019-06-06 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 82290447 |
US10561109B1 (en) | 2019-06-06 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 82042815 |
US10638687B1 (en) | 2019-06-06 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 88103087 |
US10813335B1 (en) | 2019-06-06 | 2020-10-27 | Monsanto Technology Llc | Soybean cultivar 80261309 |
US10568284B1 (en) | 2019-06-06 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 87161804 |
US10609889B1 (en) | 2019-06-06 | 2020-04-07 | Monsanto Technology Llc | Soybean cultivar 89001711 |
US10638714B1 (en) | 2019-06-06 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 84062517 |
US10555490B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 85370909 |
US10548290B1 (en) | 2019-06-07 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 89031941 |
US10568294B1 (en) | 2019-06-07 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 82440038 |
US10660296B1 (en) | 2019-06-07 | 2020-05-26 | Monsanto Technology Llc | Soybean cultivar 84241941 |
US10548274B1 (en) | 2019-06-07 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 83262718 |
US10568293B1 (en) | 2019-06-07 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar AR 86112649 |
US10561089B1 (en) | 2019-06-07 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 84060345 |
US10555491B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 83072718 |
US10602710B1 (en) | 2019-06-07 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 89282206 |
US10561090B1 (en) | 2019-06-07 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 86370345 |
US10555492B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 81430038 |
US10925238B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073320 |
US10905071B1 (en) | 2019-07-29 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072744 |
US10918054B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073353 |
US10918050B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072791 |
US10849300B1 (en) | 2019-07-29 | 2020-12-01 | Monsanto Technology Llc | Soybean variety 01073359 |
US10905072B1 (en) | 2019-07-29 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01073330 |
US10863708B1 (en) | 2019-07-29 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01073439 |
US10918052B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073365 |
US10869452B1 (en) | 2019-07-29 | 2020-12-22 | Monsanto Technology Llc | Soybean variety 01072767 |
US10939643B2 (en) | 2019-07-29 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072768 |
US10918051B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072804 |
US10932429B2 (en) | 2019-07-29 | 2021-03-02 | Monsanto Technology Llc | Soybean variety 01073265 |
US10912270B1 (en) | 2019-07-29 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01072737 |
US10918053B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073354 |
US10993401B2 (en) | 2019-07-29 | 2021-05-04 | Monsanto Technology Llc | Soybean variety 01073256 |
US10863709B1 (en) | 2019-07-29 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01072789 |
US10925237B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073288 |
US10925235B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01072810 |
US11096357B2 (en) | 2019-07-29 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01073351 |
US10932428B2 (en) | 2019-07-29 | 2021-03-02 | Monsanto Technology Llc | Soybean variety 01072749 |
US10925236B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01072823 |
US10849301B1 (en) | 2019-07-30 | 2020-12-01 | Monsanto Technology Llc | Soybean variety 01072784 |
US10939644B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073461 |
US10827714B1 (en) | 2019-07-30 | 2020-11-10 | Monsanto Technology Llc | Soybean variety 01073443 |
US10939645B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073480 |
US10939646B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073479 |
US10881074B1 (en) | 2019-07-30 | 2021-01-05 | Monsanto Technology Llc | Soybean variety 01072802 |
US10897864B1 (en) | 2019-07-30 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01073453 |
US10925240B1 (en) | 2019-07-30 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073455 |
US10925239B1 (en) | 2019-07-30 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073452 |
US10856508B1 (en) | 2019-07-30 | 2020-12-08 | Monsanto Technology Llc | Soybean variety 01073445 |
US10856507B1 (en) | 2019-07-30 | 2020-12-08 | Monsanto Technology Llc | Soybean variety 01073444 |
US11266100B2 (en) | 2019-07-30 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01073441 |
US10918055B1 (en) | 2019-07-30 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073472 |
US10888063B1 (en) | 2019-07-30 | 2021-01-12 | Monsanto Technology Llc | Soybean variety 01072800 |
US10881073B1 (en) | 2019-07-30 | 2021-01-05 | Monsanto Technology Llc | Soybean variety 01072782 |
US10973194B2 (en) | 2019-07-31 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072867 |
US10939647B2 (en) | 2019-07-31 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072860 |
US10863710B1 (en) | 2019-07-31 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01072819 |
US10842116B1 (en) | 2019-07-31 | 2020-11-24 | Monsanto Technology Llc | Soybean variety 01072832 |
US10842117B1 (en) | 2019-07-31 | 2020-11-24 | Monsanto Technology Llc | Soybean variety 01072842 |
US10827715B1 (en) | 2019-07-31 | 2020-11-10 | Monsanto Technology Llc | Soybean variety 01077799 |
US10918056B1 (en) | 2019-07-31 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01077815 |
US10874078B1 (en) | 2019-07-31 | 2020-12-29 | Monsanto Technology Llc | Soybean variety 01072817 |
US10869453B1 (en) | 2019-07-31 | 2020-12-22 | Monsanto Technology Llc | Soybean variety 01072813 |
US10939648B2 (en) | 2019-07-31 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072899 |
US10912271B1 (en) | 2019-07-31 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01072822 |
US10973193B2 (en) | 2019-07-31 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072864 |
US11122759B2 (en) | 2019-07-31 | 2021-09-21 | Monsanto Technology Llc | Soybean variety 01072870 |
US10945397B2 (en) | 2019-07-31 | 2021-03-16 | Monsanto Technology Llc | Soybean variety 01073506 |
US10959389B2 (en) | 2019-07-31 | 2021-03-30 | Monsanto Technology Llc | Soybean variety 01073503 |
US11026388B2 (en) | 2019-08-01 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01073060 |
US11019789B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01073534 |
US11006602B2 (en) | 2019-08-01 | 2021-05-18 | Monsanto Technology Llc | Soybean variety 01072946 |
US11026387B2 (en) | 2019-08-01 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01073016 |
US11129346B2 (en) | 2019-08-01 | 2021-09-28 | Monsanto Technology Llc | Soybean variety 01073533 |
US10905075B1 (en) | 2019-08-01 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01073059 |
US10925241B1 (en) | 2019-08-01 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01072974 |
US11019787B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01072951 |
US10905074B1 (en) | 2019-08-01 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072906 |
US11019788B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01072980 |
US10905073B1 (en) | 2019-08-01 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072883 |
US11096359B2 (en) | 2019-08-07 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01077377 |
US10973195B2 (en) | 2019-08-07 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01073325 |
US10980203B2 (en) | 2019-08-07 | 2021-04-20 | Monsanto Technology Llc | Soybean variety 01073305 |
US11076552B2 (en) | 2019-08-07 | 2021-08-03 | Monsanto Technology Llc | Soybean variety 01073356 |
US11019790B2 (en) | 2019-08-07 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01073304 |
US10912273B1 (en) | 2019-08-07 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01073355 |
US11096361B2 (en) | 2019-08-15 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01073072 |
US11096360B2 (en) | 2019-08-15 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01067560 |
US10952392B2 (en) | 2019-08-15 | 2021-03-23 | Monsanto Technology Llc | Soybean variety 01073151 |
US10945398B2 (en) | 2019-08-15 | 2021-03-16 | Monsanto Technology Llc | Soybean variety 01073160 |
US11044867B2 (en) | 2019-08-15 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073133 |
US11109550B2 (en) | 2019-08-15 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01073116 |
US10959390B2 (en) | 2019-08-15 | 2021-03-30 | Monsanto Technology Llc | Soybean variety 01073069 |
US11071271B2 (en) | 2019-08-15 | 2021-07-27 | Monsanto Technology Llc | Soybean variety 01073065 |
US11013196B2 (en) | 2019-08-15 | 2021-05-25 | Monsanto Technology Llc | Soybean variety 01073094 |
US11044866B2 (en) | 2019-08-15 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073114 |
US10952391B2 (en) | 2019-08-15 | 2021-03-23 | Monsanto Technology Llc | Soybean variety 01073150 |
US11044865B2 (en) | 2019-08-15 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073029 |
US10993407B2 (en) | 2019-08-15 | 2021-05-04 | Monsanto Technology Llc | Cotton variety 17R818B3XF |
US10952393B2 (en) | 2019-08-15 | 2021-03-23 | Monsanto Technology Llc | Soybean variety 01073169 |
US10905079B1 (en) | 2019-08-16 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01073335 |
US11006603B2 (en) | 2019-08-16 | 2021-05-18 | Monsanto Technology Llc | Soybean variety 01072287 |
US11044868B2 (en) | 2019-08-16 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073261 |
US10856509B1 (en) | 2019-08-16 | 2020-12-08 | Monsanto Technology Llc | Soybean variety 01073199 |
US11071272B2 (en) | 2019-08-16 | 2021-07-27 | Monsanto Technology Llc | Soybean variety 01073195 |
US10863711B1 (en) | 2019-08-16 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01073279 |
US10863712B1 (en) | 2019-08-16 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01073281 |
US10973196B2 (en) | 2019-08-16 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01073237 |
US11039590B2 (en) | 2019-08-16 | 2021-06-22 | Monsanto Technology Llc | Soybean variety 01073272 |
US10869454B1 (en) | 2019-08-16 | 2020-12-22 | Monsanto Technology Llc | Soybean variety 01073329 |
US11109551B2 (en) | 2019-08-16 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01073253 |
US11096362B2 (en) | 2019-08-16 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01073204 |
US10932430B1 (en) | 2019-08-16 | 2021-03-02 | Monsanto Technology Llc | Soybean variety 01073203 |
US11044869B2 (en) | 2019-08-16 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073270 |
US11160237B2 (en) | 2019-08-16 | 2021-11-02 | Monsanto Technology Llc | Soybean variety 01073183 |
US11129348B2 (en) | 2019-08-16 | 2021-09-28 | Monsanto Technology Llc | Soybean variety 01073219 |
US11039591B2 (en) | 2019-08-16 | 2021-06-22 | Monsanto Technology Llc | Soybean variety 01073340 |
US11129347B2 (en) | 2019-08-16 | 2021-09-28 | Monsanto Technology Llc | Soybean variety 01073210 |
US11129349B2 (en) | 2019-08-16 | 2021-09-28 | Monsanto Technology Llc | Soybean variety 01073350 |
US11266106B2 (en) | 2019-09-16 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01081407 |
US11350595B2 (en) | 2019-09-16 | 2022-06-07 | Monsanto Technology, Llc | Soybean variety 01077372 |
US11337400B2 (en) | 2019-09-16 | 2022-05-24 | Monsanto Technology, Llc | Soybean variety 01077373 |
US11185044B2 (en) | 2019-09-16 | 2021-11-30 | Monsanto Technology Llc | Soybean variety 01077343 |
US11266109B2 (en) | 2019-09-16 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01077385 |
US11266105B2 (en) | 2019-09-16 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01077338 |
US11284587B2 (en) | 2019-09-16 | 2022-03-29 | Monsanto Technology Llc | Soybean variety 01077381 |
US11178841B2 (en) | 2019-09-16 | 2021-11-23 | Monsanto Technology Llc | Soybean variety 01077386 |
US11229182B2 (en) | 2019-09-16 | 2022-01-25 | Monsanto Technology Llc | Soybean variety 01077387 |
US11259493B2 (en) | 2019-09-16 | 2022-03-01 | Monsanto Technology Llc | Soybean variety 01077341 |
US11266110B2 (en) | 2019-09-16 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01077337 |
US11185043B2 (en) | 2019-09-16 | 2021-11-30 | Monsanto Technology Llc | Soybean variety 01077334 |
US11259494B2 (en) | 2019-09-16 | 2022-03-01 | Monsanto Technology Llc | Soybean variety 01077345 |
US11266107B2 (en) | 2019-09-16 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01081408 |
US11297792B2 (en) | 2019-09-17 | 2022-04-12 | Monsanto Technology Llc | Soybean variety 01077344 |
US11266108B2 (en) | 2019-09-17 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01081409 |
US11266111B2 (en) | 2019-09-17 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01077383 |
US11576341B2 (en) | 2019-09-17 | 2023-02-14 | Monsanto Technology, Llc | Soybean variety 01072361 |
US11576340B2 (en) | 2019-09-17 | 2023-02-14 | Monsanto Technology, Llc | Soybean variety 01081414 |
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US11252922B2 (en) | 2019-10-18 | 2022-02-22 | Monsanto Technology Llc | Soybean variety 01077391 |
US11252921B2 (en) | 2019-10-18 | 2022-02-22 | Monsanto Technology Llc | Soybean variety 01077393 |
US11330777B2 (en) | 2019-10-18 | 2022-05-17 | Monsanto Technology, Llc | Soybean variety 01077390 |
US11272679B2 (en) | 2019-10-18 | 2022-03-15 | Monsanto Technology Llc | Soybean variety 01077330 |
US11330783B2 (en) | 2019-10-18 | 2022-05-17 | Monsanto Technology, Llc | Soybean variety 01077336 |
US11051483B1 (en) | 2019-12-16 | 2021-07-06 | Monsanto Technology Llc | Cotton variety 18R448B3XF |
US11154029B2 (en) | 2019-12-16 | 2021-10-26 | Monsanto Technology Llc | Cotton variety 18R445B3XF |
US11039594B1 (en) | 2019-12-16 | 2021-06-22 | Monsanto Technology Llc | Cotton variety 18R423B3XF |
US11064672B2 (en) | 2019-12-16 | 2021-07-20 | Monsanto Technology Llc | Cotton variety 18R438B3XF |
US11026392B1 (en) | 2019-12-16 | 2021-06-08 | Monsanto Technology Llc | Cotton variety 18R421B3XF |
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US11134645B2 (en) | 2020-01-24 | 2021-10-05 | Monsanto Technology Llc | Cotton variety 18R441B3XF |
US11310991B2 (en) | 2020-01-24 | 2022-04-26 | Monsanto Technology Llc | Cotton variety 18R540B3XF |
US11197454B2 (en) | 2020-01-24 | 2021-12-14 | Monsanto Technology Llc | Cotton variety 18R062 |
US11129353B2 (en) | 2020-01-24 | 2021-09-28 | Monsanto Technology Llc | Cotton variety 18R419B3XF |
US11134646B2 (en) | 2020-01-24 | 2021-10-05 | Monsanto Technology Llc | Cotton variety 18R459B3XF |
US11129352B2 (en) | 2020-01-24 | 2021-09-28 | Monsanto Technology Llc | Cotton variety 18R418B3XF |
US11129354B2 (en) | 2020-01-24 | 2021-09-28 | Monsanto Technology Llc | Cotton variety 18R435B3XF |
US11206800B2 (en) | 2020-01-24 | 2021-12-28 | Monsanto Technology Llc | Cotton variety 18R067 |
US11185046B2 (en) | 2020-01-24 | 2021-11-30 | Monsanto Technology Llc | Cotton variety 18R409B3XF |
US11154030B2 (en) | 2020-02-12 | 2021-10-26 | Monsanto Technology Llc | Cotton variety 16R020 |
US11330787B2 (en) | 2020-02-12 | 2022-05-17 | Monsanto Technology Llc | Cotton variety 18R420B3XF |
US11197455B2 (en) | 2020-02-12 | 2021-12-14 | Monsanto Technology Llc | Cotton variety 18R410B3XF |
US11234406B1 (en) | 2020-09-09 | 2022-02-01 | Monsanto Technology Llc | Soybean cultivar 98010818 |
US11252923B1 (en) | 2020-09-09 | 2022-02-22 | Monsanto Technology Llc | Soybean cultivar 99003018 |
US11206797B1 (en) | 2020-09-09 | 2021-12-28 | Monsanto Technology Llc | Soybean cultivar 97320218 |
US11234405B1 (en) | 2020-09-09 | 2022-02-01 | Monsanto Technology Llc | Soybean cultivar 90110208 |
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US11350596B2 (en) | 2020-09-11 | 2022-06-07 | Monsanto Technology, Llc | Soybean cultivar 96050600 |
US11357200B2 (en) | 2020-09-11 | 2022-06-14 | Monsanto Technology, Llc | Soybean cultivar 92131600 |
US11229183B1 (en) | 2020-09-11 | 2022-01-25 | Monsanto Technology Llc | Soybean cultivar 93070703 |
US11330784B2 (en) | 2020-09-11 | 2022-05-17 | Monsanto Technology, Llc | Soybean cultivar 95322700 |
US11277996B1 (en) | 2020-09-11 | 2022-03-22 | Monsanto Technology Llc | Soybean cultivar 97051600 |
US11284592B1 (en) | 2020-09-11 | 2022-03-29 | Monsanto Technology Llc | Soybean cultivar 91030603 |
US11310988B2 (en) | 2020-09-11 | 2022-04-26 | Monsanto Technology Llc | Soybean cultivar 92340030 |
US11317588B2 (en) | 2020-09-11 | 2022-05-03 | Stine Seed Farm, Inc. | Soybean cultivar 97080600 |
US11606926B2 (en) | 2020-09-15 | 2023-03-21 | Monsanto Technology Llc | Soybean cultivar 94030201 |
US11343988B2 (en) | 2020-09-15 | 2022-05-31 | Stine Seed Farm, Inc. | Soybean cultivar 91041800 |
US11419299B2 (en) | 2020-09-15 | 2022-08-23 | Monsanto Technology Llc | Soybean cultivar 91361201 |
US11412692B2 (en) | 2020-09-15 | 2022-08-16 | Monsanto Technology Llc | Soybean cultivar 90290600 |
US11324183B2 (en) | 2020-09-15 | 2022-05-10 | Stine Seed Farm, Inc. | Soybean cultivar 90250820 |
US11343989B2 (en) | 2020-09-15 | 2022-05-31 | Monsanto Technology, Llc | Soybean cultivar 92350300 |
US11406080B2 (en) | 2020-09-15 | 2022-08-09 | Stine Seed Farm, Inc. | Soybean cultivar 92420140 |
US11399496B2 (en) | 2020-09-15 | 2022-08-02 | Monsanto Technology, Llc | Soybean cultivar 90116100 |
US11337401B2 (en) | 2020-09-15 | 2022-05-24 | Monsanto Technology, Llc | Soybean cultivar 90370640 |
US11363778B2 (en) | 2020-09-15 | 2022-06-21 | Monsanto Technology Llc | Soybean cultivar 90221620 |
US11369074B2 (en) | 2020-09-15 | 2022-06-28 | Monsanto Technology Llc | Soybean cultivar 99122620 |
US11350597B2 (en) | 2020-09-15 | 2022-06-07 | Monsanto Technology, Llc | Soybean cultivar 94111580 |
US11357201B2 (en) | 2020-09-15 | 2022-06-14 | Monsanto Technology, Llc | Soybean cultivar 94411660 |
US11363779B2 (en) | 2020-09-15 | 2022-06-21 | Monsanto Technology Llc | Soybean cultivar 90301201 |
US11425879B2 (en) | 2020-09-15 | 2022-08-30 | Monsanto Technology, Llc | Soybean cultivar 93054930 |
US11343987B2 (en) | 2020-09-15 | 2022-05-31 | Stine Seed Farm, Inc. | Soybean cultivar 90401320 |
US11310989B2 (en) | 2020-09-15 | 2022-04-26 | Monsanto Technology Llc | Soybean cultivar 94071660 |
US11363780B2 (en) | 2020-09-15 | 2022-06-21 | Monsanto Technology Llc | Soybean cultivar 93272026 |
US11399497B2 (en) | 2020-09-15 | 2022-08-02 | Monsanto Technology, Llc | Soybean cultivar 93333110 |
US11744211B2 (en) | 2020-09-15 | 2023-09-05 | Monsanto Technology, Llc | Soybean cultivar 92310017 |
US11324182B2 (en) | 2020-09-15 | 2022-05-10 | Stine Seed Farm, Inc. | Soybean cultivar 95060020 |
US11317589B2 (en) | 2020-09-15 | 2022-05-03 | Monsanto Technology Llc | Soybean cultivar 95430820 |
US11324184B2 (en) | 2020-09-15 | 2022-05-10 | Stine Seed Farm, Inc. | Soybean cultivar 97061850 |
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US11653613B2 (en) | 2020-11-02 | 2023-05-23 | Monsanto Technology Llc | Soybean variety 01077829 |
US11477960B2 (en) | 2020-11-02 | 2022-10-25 | Monsanto Technology Llc | Soybean variety 01077912 |
US11653612B2 (en) | 2020-11-02 | 2023-05-23 | Monsanto Technology Llc | Soybean variety 01077826 |
US11547075B2 (en) | 2020-11-02 | 2023-01-10 | Monsanto Technology Llc | Soybean variety 01078822 |
US11596122B2 (en) | 2020-11-02 | 2023-03-07 | Monsanto Technology Llc | Soybean variety 01077891 |
US11510382B2 (en) | 2020-11-02 | 2022-11-29 | Monsanto Technology Llc | Soybean variety 01078845 |
US11412694B2 (en) | 2020-11-05 | 2022-08-16 | Monsanto Technology Llc | Soybean variety 01077905 |
US11457591B2 (en) | 2020-11-05 | 2022-10-04 | Monsanto Technology, Llc | Soybean variety 01077893 |
US11357205B2 (en) | 2020-11-05 | 2022-06-14 | Monsanto Technology Llc | Soybean variety 01083631 |
US11343991B2 (en) | 2020-11-05 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01078115 |
US11445688B2 (en) | 2020-11-05 | 2022-09-20 | Monsanto Technology, Llc | Soybean variety 01077870 |
US11343990B2 (en) | 2020-11-05 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01083439 |
US11343993B2 (en) | 2020-11-05 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01083650 |
US11350600B2 (en) | 2020-11-05 | 2022-06-07 | Monsanto Technology, Llc | Soybean variety 01083699 |
US11533883B2 (en) | 2020-11-05 | 2022-12-27 | Monsanto Technology, Llc | Soybean variety 01077890 |
US11284593B1 (en) | 2020-11-05 | 2022-03-29 | Monsanto Technology Llc | Soybean variety 01083687 |
US11343992B2 (en) | 2020-11-05 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01078550 |
US11445689B2 (en) | 2020-11-05 | 2022-09-20 | Monsanto Technology, Llc | Soybean variety 01078100 |
US11277997B1 (en) | 2020-11-05 | 2022-03-22 | Monsanto Technology Llc | Soybean variety 01077853 |
US11284594B1 (en) | 2020-11-05 | 2022-03-29 | Monsanto Technology Llc | Soybean variety 01077848 |
US11432518B2 (en) | 2020-11-06 | 2022-09-06 | Monsanto Technology, Llc | Soybean variety 01078759 |
US11484000B2 (en) | 2020-11-06 | 2022-11-01 | Monsanto Technology, Llc | Soybean variety 01083661 |
US11547077B2 (en) | 2020-11-06 | 2023-01-10 | Monsanto Technology, Llc | Soybean variety 01083688 |
US11330785B1 (en) | 2020-11-06 | 2022-05-17 | Monsanto Technology, Llc | Soybean variety 01077941 |
US11343996B1 (en) | 2020-11-06 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01077875 |
US11439114B2 (en) | 2020-11-06 | 2022-09-13 | Monsanto Technology, Llc | Soybean variety 01078032 |
US11350601B2 (en) | 2020-11-06 | 2022-06-07 | Monsanto Technology, Llc | Soybean variety 01077876 |
US11457601B2 (en) | 2020-11-06 | 2022-10-04 | Monsanto Technology Llc | Soybean variety 01083638 |
US11477961B2 (en) | 2020-11-06 | 2022-10-25 | Monsanto Technology, Llc | Soybean variety 01077940 |
US11510384B2 (en) | 2020-11-06 | 2022-11-29 | Monsanto Technology, Llc | Soybean variety 01078675 |
US11528879B2 (en) | 2020-11-06 | 2022-12-20 | Monsanto Technology, Llc | Soybean variety 01083659 |
US11457600B2 (en) | 2020-11-06 | 2022-10-04 | Monsanto Technology, Llc | Soybean variety 01083646 |
US11382301B2 (en) | 2020-11-06 | 2022-07-12 | Monsanto Technology, Llc | Soybean variety 01077881 |
US11553671B2 (en) | 2020-11-06 | 2023-01-17 | Monsanto Technology, Llc | Soybean variety 01083653 |
US11533884B2 (en) | 2020-11-06 | 2022-12-27 | Monsanto Technology, Llc | Soybean variety 01083660 |
US11343994B1 (en) | 2020-11-06 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01083655 |
US11547076B2 (en) | 2020-11-06 | 2023-01-10 | Monsanto Technology, Llc | Soybean variety 01077931 |
US11445690B2 (en) | 2020-11-06 | 2022-09-20 | Monsanto Technology Llc | Soybean variety 01083639 |
US11369076B2 (en) | 2020-11-06 | 2022-06-28 | Monsanto Technology, Llc | Soybean variety 01078749 |
US11464195B2 (en) | 2020-11-06 | 2022-10-11 | Monsanto Technology Llc | Soybean variety 01083654 |
US11343995B1 (en) | 2020-11-06 | 2022-05-31 | Monsanto Technology, Llc | Soybean variety 01083647 |
US11357206B2 (en) | 2020-11-06 | 2022-06-14 | Monsanto Technology Llc | Soybean variety 01077974 |
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