CN110591924B - 一种羊肚菌菌株及其应用 - Google Patents
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Abstract
本发明公开了一种羊肚菌菌株,该羊肚菌菌株命名为粗柄羊肚菌OL‑Y‑B(Morchella crassepsis strain OL‑Y‑B),在中国微生物菌种保藏管理委员会普通微生物中心的保藏号为CGMCC No.18115。该粗柄羊肚菌OL‑Y‑B通过发酵培养得到的发酵产物菌悬液可代替化肥施用,并可显著促进宁南固原地区糜子生长。对玉米、小麦、荞麦等谷类作物也有明显促进作用,并可提高小麦耐盐性。该粗柄羊肚菌OL‑Y‑B的获得,有望为宁南黄土高原地区糜子的“双减”发挥积极作用,具有良好的应用前景。
Description
技术领域
本发明涉及微生物及其应用领域,涉及一种菌株,特别是一种羊肚菌菌株及其应用。
背景技术
植物内生菌是指生活史的一定阶段或全部阶段生活于健康植物的各种组织和器官内部的微生物。内生菌因溶磷、固氮、产生植物激素、诱导抗虫抗病、抗逆等强大生理和生态功能而倍受关注。近年来发现,许多植物内生菌还能够产生具有农药活性的次生代谢产物,引起了人们巨大的兴趣。欧盟在 2012年起明确提出谷类作物绝对不使用化肥的禁令,中国农业部近年来也提出农业生产“双减”口号,寻找新型农药制剂和肥料已成为国内外共同的发展趋势,内生菌、菌根菌等益微生物的高效利用无疑成为现代农业发展的突破口。
糜子(Panicum miliaceum L.),禾本科黍属(Panicum miliaceum),在我国栽培历史悠久,具有生育期短、抗逆性强等特点,在甘肃、宁夏、内蒙古、山西、陕西等省区均有种植,全国总种植面积约53.3万hm2,是这些地区主要粮食作物和经济作物,在旱区种植业结构调整和产业发展中占有重要地位。同时,糜子因其籽粒中富含多种有益人体的营养物质,具有较高的食用价值和药用价值,越来越受到消费者的青睐,市场需求量日益增长。
但在糜子生产过程中,由于受贫瘠的土壤条件以及气候条件等诸多环境因子的限制,糜子单产水平普遍偏低,较低的单产水平限制宁南地区糜子产业的发展。因此,通过利用内生菌、菌根菌等益微生物对糜子产生的促生作用,改善糜子在低施氮水平条件、干旱条件以及零农药使用条件下的生长情况,保障糜子的单产水平,对于推动糜子产业发展以及实现可持续农业具有重要意义。
发明内容
本发明的目的在于,提供一种羊肚菌菌株,该羊肚菌菌株通过发酵培养得到的发酵产物菌悬液可代替化肥施用,并通过试验证明该羊肚菌菌株能够用于促进糜子、玉米、小麦、荞麦共生的应用。
为了实现上述任务,本发明采取如下的技术解决方案:
一种羊肚菌菌株,其特征在于,该羊肚菌菌株命名为粗柄羊肚菌OL-Y-B(Morchella crassepsis strain OL-Y-B),在中国微生物菌种保藏管理委员会普通微生物中心的保藏号为CGMCC No.18115。
经测定,该粗柄羊肚菌OL-Y-B的ITS-nrDNA序列如下所示:
GAGAGAGAGTCAGCTACGATGGGGCGCGCCGGCAAACCAGACCGCTGGCACACTGGTGGC GGGTACTGTCACCCGCGCCGTTTGTCGCAACGTCCAGGGCGAGAGCCTTAGCCGGGTGTGGCCC CATCCGTTAAGCTAGTCAATTCCATGCTGCCTGCTGCGCTCCCTCCCCACGCCCTGGGCAGCAG CTCCCGCAATCGGGGGTGGAAGCCGGATCAAGGAATCAATGCTCTGCATCCATTCAGTCAATTG AATGTGGTGCAGTTTAGGTTATTTCTGCTGAAGGGGGATCCCCCGGCAGAGGTGGACGGCACAT GCAGGCTCCGAACGACGGAAGCCAACCATGATGCTGAGTCCCCTGGGGCTAGGGGTGGTAAAAG CCCCTAGCCACTCCCCACCGACGGCGTTGACCCCATCGTTCTGGCACCCTCTCGCCACTGTCCC AACCAAAACCCTCTGTGTACCCTTCCCTGTTGCTTCCCCCGGGAAACTGGCTCCGGCCAGCCGG GGGGAGAAACCAAGCAAAAACCCTTTTCGCAAAACAGACGTCTGAACGCAAAAAACAAAACAAA AAAAAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCCACATCGATGAAGAACGCAGCGAAAT GCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCC TCTGGTATTCCGGAGGGCATGCCTGTTCGAGCGTCATAAATACCGCTCCCCCTCGGATTGCTTG CGGTCCCTGGGGGGTTCTGGCAATGTGGATTCCCCGTGCTTTGAGGGCATGCGAACGGGCTCCC AGTGCTGAAAGACATAATGTTCCCAGCCGAAACCGGTGATTAATTTCATCGGCAGGATTCGTGGCAGGCAGACTGAGGGCGTCAACCGTGGAGTCATGAGGGTAGAAACCTCCCCCTTTGCAAGTAAC ATTGCTCTGGCAGTTAGATCTGCAGGCCTGCCGGTCTGGGGATGGACCTCCCACTCGCAGGCGT CACGGCCACGATAGCGGGCGTTAAACGGAATCCGATCCGCCCTCACGGGTTGGTTGAAGATCCT TGGGGGCTAG。
该粗柄羊肚菌OL-Y-B的LSU-nrDNA序列如下所示:
CCATGCCCATATTTGACGATCGATTTGCACGTCAGAACCGCTGCGAGCCTCCACCAGAGTTT CCTCTGGCTTCACCCTATACAGGCATAGTTCACCTTCTTTCGGGTCCGGCCCCGTATGCTCTTACT CAAATCCATCCGAGAACATCAGGATCGGTCGGTGATGCGCCGAAGCTCTCACCTACGTTCACTTTC ATTACGCGTAGGGGTTTGACACCCGAACACTCGCATACGAAGACGACTCCTTGGTCCGTGTTTCAA GACGGGTCATTGATGACCATTACGCCAGCATCCGTGCCGAAGCGCGTTCCTCAGTCCGCCCCAGGG CATTACACGACGGGCTATAACACTCCCCGAAGGGAGCCACATTCCCGCCGCCTTTATCCCCCGGGG CAAACTGATGCTGGCCTGGACTGGCAGAGTGCACCGGCGAGAACGCCGGATGTTCAACCAAGCCCA AGTCTGGTCATGAATGCTTCCCTTTCAACAATTTCACGTACTTTTTAACCCTCTTTTCAAAGTGCT TTTCATCTTTCGATCACTCTACTTGTGCGCTATCGGTCTCTGGCCGGTATTTAGCTTTAGAAGACG TACACCTCCCATTTAGAGCAGCATTCCCAAACTACTCGACTCGTCGAAGGAGCTTTACAGAGGCTT GGTGGTCCTACCGTACGGGGCTCTCACCCTCTATGGCGTCCCGTTCCAGGGAACTCGGAAGGCACC TCGCCAAAAGCATCCTCTACAAATTACAACTCGGACCTTGCGGCCAGATTTCAAATTTGAGCTGTT GCCGCTTCACTCGCCGTTACTGAGGCAATCCCTGTTGGTTTCTTTTCCTCCGCTTA。
该粗柄羊肚菌OL-Y-B的EF基因序列如下所示:
TGATCTACAAGGTTTGCTCTCGATCTTGGTGAATAGAAAAAAAGAAAAAAAGAAAACTGA CTGAATTTTTCTACCATATCAGTGTGGTGGTATTGACAAGCGTACCATCGAGAAGTTCGAGAAG GAGGCTGCTGAACTCGGAAAGGGTTCTTTCAAGTACGCCTGGGTTTTGGACAAATTGAAGGCCG AGCGTGAGCGTGGTATTACCATCGATATCGCCCTCTGGAAGTTCGAGACCCCTAAGTACTATGT CACTGTCATTGGTAAGATAGCCCCGTACCTTATCTTTTTAATCTTTGTGTATCTCGATCTACCC CGCTAGTCCGGTTTGAAATTTTCCCGGGACAACGCTAATACTTCTTTTAGATGCCCCTGGTCAT CGTGATTTCATCAAGAACATGATCACTGGTACTTCCCAGGCTGATTGTGCCGTTCTCATTATTG CCGCTGGTGTAAGTTATTGTCATACTGTCTATTTTTTTATGATTATGTTCTAATTGCTCTACAA AATAGACTGGTGAGTTCGAGGCTGGTATCTCCAAGGATGGTCAGACCCGTGAGCACGCCCTTCT CGCCTACACCCTTGGTGTTAAGCAGCTCATTGTTGCTATCAACAAGATGGACACCACCAAGTGG TCTGAGAATCGTTT。
该粗柄羊肚菌OL-Y-B的RBP1基因序列如下所示:
ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTA AGAAAGTTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACAC TGAATAAGTGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAAC CCTCAATTTGCGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTC TGTGCAAAGCAAAATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGG CGATGACGCAGGCACCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCT ACTATCAGGCGTGAAGGTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGG AAACCACTCAGGGAGAGAAAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACAT CACCAGTGATGAAATTAGGAAAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTG ATTACTGTTCTTCCCGTGCCGCCGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTG GAGGAATGCGTGGGGAGGACGATTTGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAG TGTACGCGATGCCGAGCAAAACGGATCTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTG CAGTTTCAT。
该粗柄羊肚菌OL-Y-B的RBP2基因序列如下所示:
AAGGGAATGATACTGGCGCAGATACCCAAGATCATACTGGGGTGGATCTCACAATGTGTCCA TGTGTGCGCAGTGTGGGGGTTGATCGGCGCCTTGACACGCTTTGCGCTGAACTCATCTTTGGGCTC CTCGTAACCCAGCTGCATCTGCCTACTCATATCCAGATCCTCGGGCGTCATGACGATCATGACCGT CTCCTCTTCCTCTGCATCCAGGTACTCCACGGTACCACTAAACAGAAGACCTTCCCACCCGAACCG GTCCTCCATCGGCAGCGCATGATCGTCTTCCAACTTCTCGATATGCTCCTTCTGGAGCACCAGCTG TCCTCGCCTCTCACTTCGCGGGTCATTGTCAATGACGAACAACGGCCTACATACACGCCCAGCATC TGTAAAAATCTTGAACTCCCTGTCACGAATATCCCTAATGATGGAAACTTCATGGTTGACAACACC CTGCCTTCTTACGTCTTGGACAACCTTGTGCAACATGAGCGGGTCACGATGCGTTCCGACCCAGAC ACCATTGACAAATACCTTTGTCGCATTCGGCGATCTCTGCGGCTCATACTCCTCCAGAAGCTCCAT ATCATTCGAAGATAAAAAGTCCACGAGCGGGTATCCGGGCGTTCCGATTGTGATGTAGGACATGAG TGACAAGTTCTTAACGAGACCACACGCCTGTCCCTCGGGC。
上述粗柄羊肚菌OL-Y-B的培养条件是:在黑暗条件下,采用pH 7.0-8.5 的PDA培养基,培养温度为25℃-30℃。
经申请人的试验证明,本发明得到的羊肚菌菌株,通过发酵培养得到的发酵产物菌悬液可代替化肥施用,可以用于促进糜子、米、小麦或荞麦共生的应用。尤其是对宁南固原地区糜子的萌发率、苗高、产量、主穗粒重、主穗重、千粒重等等均有良好的促进作用,有望为宁南黄土高原地区糜子的“双减”发挥积极作用。
经申请人验证,粗柄羊肚菌OL-Y-B发酵后的发酵液,含有ABA、SA、IAA 等植物激素,对谷类作物具有促生抗逆功效。具有良好的应用前景。
所述的粗柄羊肚菌OL-Y-B经发酵培养后的发酵液菌体和PDA平板培养产生的菌核和菌丝体,用于替代化肥制备促生长的生物制剂,所述的发酵培养条件为:PD液体培养基,pH值7.0-8.0,转速80-150rpm,25-30℃;PDA 固体培养基,pH值7.0-8.0,黑暗培养2周。
本发明的粗柄羊肚菌OL-Y-B的获得,有望为宁南黄土高原地区糜子的“双减”发挥积极作用,具有良好的应用前景。
附图说明
图1是粗柄羊肚菌OL-Y-B形态和***发育分析,其中,图1-1是粗柄羊肚菌OL-Y-B形态(A图是粗柄羊肚菌OL-Y-B,B图为子囊,C图为子实层),图1-2是ITS-nrDNA***发育树,图1-3是5基因联合***树。
图2是粗柄羊肚菌OL-Y-B对糜子的田间促生作用,其中,a图是盆栽糜子图片,b是田间糜子图片。
图3是粗柄羊肚菌OL-Y-B与糜子的共生关系。
图4是粗柄羊肚菌OL-Y-B与玉米、小米、荞麦的共生关系。其中,图4-1 是粗柄羊肚菌OL-Y-B与玉米的共生关系,图4-2是粗柄羊肚菌OL-Y-B与小米的共生关系,图4-3是粗柄羊肚菌OL-Y-B与荞麦的共生关系。
图5是粗柄羊肚菌OL-Y-B对玉米的促生作用。
图6是粗柄羊肚菌OL-Y-B对冬小麦的耐盐促生作用。
图7是粗柄羊肚菌OL-Y-B发酵液植物激素含量。
以下结合附图和实施例对本发明作进一步的详细说明。
具体实施方式
实施例1:粗柄羊肚菌OL-Y-B的筛选及其鉴定
1、申请人的课题组在中国陕西省凤县烧荒地采集到一株菌株,采集具体方法为:从子囊果上取新鲜的子囊盘贴于灭菌的培养皿上盖并密封培养皿,利用PDA培养基分离、培养、纯化,结果得到能大量产生菌核的菌株,申请人将其命名为粗柄羊肚菌OL-Y-B(Morchella crassipes strain OL-Y-B,以下简称菌株OL-Y-B)。
经过申请人的实验表明,该菌株OL-Y-B对糜子、玉米等具有促生作用,实验结果如图2、图3、图5和图6所示。
2、菌株OL-Y-B的种属鉴定
参见图1,PDA培养基上,菌落初期乳白色,菌丝粗、放射状生长,气丝不发达(图1-1的B图)。1周后,菌落逐渐变黄色,菌核开始出现(图1-1 的C图);2周后,菌核大量产生,主要分布在菌落中央,散生,突出,大小 1-2mm。菌株OL-Y-B喜微碱性、含氮少的培养基。耐高温,最高生长温度可达45℃,最低生长温度10℃,最适生长温度25-28℃。
采用常规形态学及多基因分子***学对菌株OL-Y-B进行鉴定,确定该菌株OL-Y-B属于粗柄羊肚菌。
以菌株OL-Y-B的基因组DNA为模板,以18SrDNA、ITS、EF、RBP1、RBP2 序列通用引物进行扩增,PCR产物经检测后送上海生工生物工程有限公司测序,经测定,得到如下序列:
1)菌株OL-Y-B的ITS-nrDNA序列如下所示:
GAGAGAGAGTCAGCTACGATGGGGCGCGCCGGCAAACCAGACCGCTGGCACACTGGTGGC GGGTACTGTCACCCGCGCCGTTTGTCGCAACGTCCAGGGCGAGAGCCTTAGCCGGGTGTGGCCC CATCCGTTAAGCTAGTCAATTCCATGCTGCCTGCTGCGCTCCCTCCCCACGCCCTGGGCAGCAG CTCCCGCAATCGGGGGTGGAAGCCGGATCAAGGAATCAATGCTCTGCATCCATTCAGTCAATTG AATGTGGTGCAGTTTAGGTTATTTCTGCTGAAGGGGGATCCCCCGGCAGAGGTGGACGGCACAT GCAGGCTCCGAACGACGGAAGCCAACCATGATGCTGAGTCCCCTGGGGCTAGGGGTGGTAAAAG CCCCTAGCCACTCCCCACCGACGGCGTTGACCCCATCGTTCTGGCACCCTCTCGCCACTGTCCC AACCAAAACCCTCTGTGTACCCTTCCCTGTTGCTTCCCCCGGGAAACTGGCTCCGGCCAGCCGG GGGGAGAAACCAAGCAAAAACCCTTTTCGCAAAACAGACGTCTGAACGCAAAAAACAAAACAAA AAAAAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCCACATCGATGAAGAACGCAGCGAAAT GCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCC TCTGGTATTCCGGAGGGCATGCCTGTTCGAGCGTCATAAATACCGCTCCCCCTCGGATTGCTTG CGGTCCCTGGGGGGTTCTGGCAATGTGGATTCCCCGTGCTTTGAGGGCATGCGAACGGGCTCCC AGTGCTGAAAGACATAATGTTCCCAGCCGAAACCGGTGATTAATTTCATCGGCAGGATTCGTGGCAGGCAGACTGAGGGCGTCAACCGTGGAGTCATGAGGGTAGAAACCTCCCCCTTTGCAAGTAAC ATTGCTCTGGCAGTTAGATCTGCAGGCCTGCCGGTCTGGGGATGGACCTCCCACTCGCAGGCGT CACGGCCACGATAGCGGGCGTTAAACGGAATCCGATCCGCCCTCACGGGTTGGTTGAAGATCCT TGGGGGCTAG。
2)菌株OL-Y-B的LSU-nrDNA序列如下所示:
CCATGCCCATATTTGACGATCGATTTGCACGTCAGAACCGCTGCGAGCCTCCACCAGAGTTT CCTCTGGCTTCACCCTATACAGGCATAGTTCACCTTCTTTCGGGTCCGGCCCCGTATGCTCTTACT CAAATCCATCCGAGAACATCAGGATCGGTCGGTGATGCGCCGAAGCTCTCACCTACGTTCACTTTC ATTACGCGTAGGGGTTTGACACCCGAACACTCGCATACGAAGACGACTCCTTGGTCCGTGTTTCAA GACGGGTCATTGATGACCATTACGCCAGCATCCGTGCCGAAGCGCGTTCCTCAGTCCGCCCCAGGG CATTACACGACGGGCTATAACACTCCCCGAAGGGAGCCACATTCCCGCCGCCTTTATCCCCCGGGG CAAACTGATGCTGGCCTGGACTGGCAGAGTGCACCGGCGAGAACGCCGGATGTTCAACCAAGCCCA AGTCTGGTCATGAATGCTTCCCTTTCAACAATTTCACGTACTTTTTAACCCTCTTTTCAAAGTGCT TTTCATCTTTCGATCACTCTACTTGTGCGCTATCGGTCTCTGGCCGGTATTTAGCTTTAGAAGACG TACACCTCCCATTTAGAGCAGCATTCCCAAACTACTCGACTCGTCGAAGGAGCTTTACAGAGGCTT GGTGGTCCTACCGTACGGGGCTCTCACCCTCTATGGCGTCCCGTTCCAGGGAACTCGGAAGGCACC TCGCCAAAAGCATCCTCTACAAATTACAACTCGGACCTTGCGGCCAGATTTCAAATTTGAGCTGTT GCCGCTTCACTCGCCGTTACTGAGGCAATCCCTGTTGGTTTCTTTTCCTCCGCTTA。
3)菌株OL-Y-B的EF基因序列如下所示:
TGATCTACAAGGTTTGCTCTCGATCTTGGTGAATAGAAAAAAAGAAAAAAAGAAAACTGA CTGAATTTTTCTACCATATCAGTGTGGTGGTATTGACAAGCGTACCATCGAGAAGTTCGAGAAG GAGGCTGCTGAACTCGGAAAGGGTTCTTTCAAGTACGCCTGGGTTTTGGACAAATTGAAGGCCG AGCGTGAGCGTGGTATTACCATCGATATCGCCCTCTGGAAGTTCGAGACCCCTAAGTACTATGT CACTGTCATTGGTAAGATAGCCCCGTACCTTATCTTTTTAATCTTTGTGTATCTCGATCTACCC CGCTAGTCCGGTTTGAAATTTTCCCGGGACAACGCTAATACTTCTTTTAGATGCCCCTGGTCAT CGTGATTTCATCAAGAACATGATCACTGGTACTTCCCAGGCTGATTGTGCCGTTCTCATTATTG CCGCTGGTGTAAGTTATTGTCATACTGTCTATTTTTTTATGATTATGTTCTAATTGCTCTACAA AATAGACTGGTGAGTTCGAGGCTGGTATCTCCAAGGATGGTCAGACCCGTGAGCACGCCCTTCT CGCCTACACCCTTGGTGTTAAGCAGCTCATTGTTGCTATCAACAAGATGGACACCACCAAGTGG TCTGAGAATCGTTT。
4)菌株OL-Y-B的RBP1基因序列如下所示:
ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTA AGAAAGTTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACAC TGAATAAGTGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAAC CCTCAATTTGCGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTC TGTGCAAAGCAAAATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGG CGATGACGCAGGCACCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCT ACTATCAGGCGTGAAGGTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGG AAACCACTCAGGGAGAGAAAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACAT CACCAGTGATGAAATTAGGAAAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTG ATTACTGTTCTTCCCGTGCCGCCGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTG GAGGAATGCGTGGGGAGGACGATTTGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAG TGTACGCGATGCCGAGCAAAACGGATCTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTG CAGTTTCAT。
5)菌株OL-Y-B的RBP2基因序列如下所示:
AAGGGAATGATACTGGCGCAGATACCCAAGATCATACTGGGGTGGATCTCACAATGTGTCCA TGTGTGCGCAGTGTGGGGGTTGATCGGCGCCTTGACACGCTTTGCGCTGAACTCATCTTTGGGCTC CTCGTAACCCAGCTGCATCTGCCTACTCATATCCAGATCCTCGGGCGTCATGACGATCATGACCGT CTCCTCTTCCTCTGCATCCAGGTACTCCACGGTACCACTAAACAGAAGACCTTCCCACCCGAACCG GTCCTCCATCGGCAGCGCATGATCGTCTTCCAACTTCTCGATATGCTCCTTCTGGAGCACCAGCTG TCCTCGCCTCTCACTTCGCGGGTCATTGTCAATGACGAACAACGGCCTACATACACGCCCAGCATC TGTAAAAATCTTGAACTCCCTGTCACGAATATCCCTAATGATGGAAACTTCATGGTTGACAACACC CTGCCTTCTTACGTCTTGGACAACCTTGTGCAACATGAGCGGGTCACGATGCGTTCCGACCCAGAC ACCATTGACAAATACCTTTGTCGCATTCGGCGATCTCTGCGGCTCATACTCCTCCAGAAGCTCCAT ATCATTCGAAGATAAAAAGTCCACGAGCGGGTATCCGGGCGTTCCGATTGTGATGTAGGACATGAG TGACAAGTTCTTAACGAGACCACACGCCTGTCCCTCGGGC。
根据ITS-nrDNA***发育树和5基因联合***树分析表明,菌株OL-Y-B 为Morchella crassispes Mes-9(图1)。
该菌株OL-Y-B,已于2019年08月02日提交中国微生物菌种保藏管理委员会普通微生物中心,保藏地址:中国北京,保藏号为CGMCC No.18115,根据中国微生物菌种保藏管理委员会普通微生物中心检测,结果为存活。
实施例2:菌株OL-Y-B的室内培养及产生植物生长激素特征
该菌株OL-Y-B的种子培养条件为:
PDA培养基(pH 7.0-8.5),培养温度25-30℃,黑暗条件。
该菌株OL-Y-B的发酵液培养条件为:
PD液体培养基,pH7.6,转速80rpm,25-30℃。
以菌株OL-Y-B的发酵一液,经细菌过滤器灭菌后,用WondasilTM,C18 (5um,4.6×150mm;Shimadzu)柱子进行分离,采用LC-20AD(Shimadzu, Kinh Do,Japan)和电喷雾串联质谱仪(API 2000TM,Allen-Bradley,Milwaukee, WI,USA),进行植物激素种类和含量的测量。
测定结果表明,菌株OL-Y-B发酵液含有大量的水杨酸SA,其次是生长激素IAA和脱落酸ABA,无其他植物激素含量(参见图7),可代替化肥施用。
实施例3:菌株OL-Y-B对糜子萌发作用的评价
该菌株OL-Y-B在PDA平板2周后,用无菌刮铲刮取菌落表面菌核,制作的菌悬液(100mL/皿),用菌悬液液黑暗条件下室温侵泡糜子种子24h,糜子种子盆栽和田间播种(图2),对照可提高糜子萌发率12.3%和4.56%。
实施例4:菌株Y-B对糜子田间作用的评价
经菌株OL-Y-B悬液处理的糜子,田间播种结果显示,菌株OL-Y-B悬液对糜子株高、穗长、穗茎长、草重、主穗粒重、主穗重、千粒重都有提高,除穗长、株高无显著差异外,其他指标均有显著的提高;菌株OL-Y-B悬液较对照处理的其他菌剂表现更优良的性状,如下表1-1、1-2所示。
表1-1:不同生物制剂对糜子主穗粒重、主穗重、千粒重的影响
注:表中OL-Y-B即为菌株OL-Y-B悬液。
表1-2:不同生物制剂对糜子株高、穗长、穗茎长、草重的影响
注:表中OL-Y-B即为菌株OL-Y-B悬液。
实施例5:菌株OL-Y-B对盆栽玉米的促生作用的评价
对陕丹“985”玉米测试表明,分别在苗期10d,20d,30d,菌株OL-Y-B 悬液通过灌苗处理,能够促进玉米对土壤营养物质的吸收和提高玉米抗氧化酶活性,改善玉米光合作用。能成功定植玉米根部并建立共生关系(图4),当剂量为100mL和150mL时(浓度:1皿/100mL),较对照(常规管理)和尿素处理,均可显著提高玉米的生物量,根生长量、茎粗度、旗叶的面积,较对照和尿素组均有显著提高(图5)。
实施例6:菌株OL-Y-B对盆栽冬小麦的促生作用的评价
菌株OL-Y-B和木炭、玉米芯等的耦合剂培养物1.5%、2.0%、2.5%添加到含2%NaCl的土壤中,可显著提高冬小麦小偃6号根/茎比率,从而提高冬小麦的耐盐能力和生长量(见表2和图6)。
菌株OL-Y-B可和冬小麦建立共生关系(图4)。
表2:菌株OL-Y-B对冬小麦生长量和根/茎比率的作用(P<0.05)
核苷酸序列表
<110>西北农林科技大学
<120>一种羊肚菌菌株及其应用
<160>
<210>1
<211>1094
<212>粗柄羊肚菌OL-Y-B的ITS-nrDNA序列
<213>DNA
<220>
<400>
GAGAGAGAGTCAGCTACGATGGGGCGCGCCGGCAAACCAGACCGCTGGCACACTGGTGGCGGGTAC TGTCACCCGCGCCGTTTGTCGCAACGTCCAGGGCGAGAGCCTTAGCCGGGTGTGGCCCCATCCGTT AAGCTAGTCAATTCCATGCTGCCTGCTGCGCTCCCTCCCCACGCCCTGGGCAGCAGCTCCCGCAAT CGGGGGTGGAAGCCGGATCAAGGAATCAATGCTCTGCATCCATTCAGTCAATTGAATGTGGTGCAG TTTAGGTTATTTCTGCTGAAGGGGGATCCCCCGGCAGAGGTGGACGGCACATGCAGGCTCCGAACG ACGGAAGCCAACCATGATGCTGAGTCCCCTGGGGCTAGGGGTGGTAAAAGCCCCTAGCCACTCCCC ACCGACGGCGTTGACCCCATCGTTCTGGCACCCTCTCGCCACTGTCCCAACCAAAACCCTCTGTGT ACCCTTCCCTGTTGCTTCCCCCGGGAAACTGGCTCCGGCCAGCCGGGGGGAGAAACCAAGCAAAAA CCCTTTTCGCAAAACAGACGTCTGAACGCAAAAAACAAAACAAAAAAAAGTTAAAACTTTCAACAA CGGATCTCTTGGTTCCCACATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGA ATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGTATTCCGGAGGGCATGCCTGT TCGAGCGTCATAAATACCGCTCCCCCTCGGATTGCTTGCGGTCCCTGGGGGGTTCTGGCAATGTGG ATTCCCCGTGCTTTGAGGGCATGCGAACGGGCTCCCAGTGCTGAAAGACATAATGTTCCCAGCCGA AACCGGTGATTAATTTCATCGGCAGGATTCGTGGCAGGCAGACTGAGGGCGTCAACCGTGGAGTCA TGAGGGTAGAAACCTCCCCCTTTGCAAGTAACATTGCTCTGGCAGTTAGATCTGCAGGCCTGCCGG TCTGGGGATGGACCTCCCACTCGCAGGCGTCACGGCCACGATAGCGGGCGTTAAACGGAATCCGAT CCGCCCTCACGGGTTGGTTGAAGATCCTTGGGGGCTAG。
<210>2
<211>844
<212>粗柄羊肚菌OL-Y-B的LSU-nrDNA序列
<213>DNA
<220>
<400>
CCATGCCCATATTTGACGATCGATTTGCACGTCAGAACCGCTGCGAGCCTCCACCAGAGTTTCCTC TGGCTTCACCCTATACAGGCATAGTTCACCTTCTTTCGGGTCCGGCCCCGTATGCTCTTACTCAAA TCCATCCGAGAACATCAGGATCGGTCGGTGATGCGCCGAAGCTCTCACCTACGTTCACTTTCATTA CGCGTAGGGGTTTGACACCCGAACACTCGCATACGAAGACGACTCCTTGGTCCGTGTTTCAAGACG GGTCATTGATGACCATTACGCCAGCATCCGTGCCGAAGCGCGTTCCTCAGTCCGCCCCAGGGCATT ACACGACGGGCTATAACACTCCCCGAAGGGAGCCACATTCCCGCCGCCTTTATCCCCCGGGGCAAA CTGATGCTGGCCTGGACTGGCAGAGTGCACCGGCGAGAACGCCGGATGTTCAACCAAGCCCAAGTC TGGTCATGAATGCTTCCCTTTCAACAATTTCACGTACTTTTTAACCCTCTTTTCAAAGTGCTTTTC ATCTTTCGATCACTCTACTTGTGCGCTATCGGTCTCTGGCCGGTATTTAGCTTTAGAAGACGTACA CCTCCCATTTAGAGCAGCATTCCCAAACTACTCGACTCGTCGAAGGAGCTTTACAGAGGCTTGGTG GTCCTACCGTACGGGGCTCTCACCCTCTATGGCGTCCCGTTCCAGGGAACTCGGAAGGCACCTCGC CAAAAGCATCCTCTACAAATTACAACTCGGACCTTGCGGCCAGATTTCAAATTTGAGCTGTTGCCG CTTCACTCGCCGTTACTGAGGCAATCCCTGTTGGTTTCTTTTCCTCCGCTTA。
<210>3
<211>773
<212>粗柄羊肚菌OL-Y-B的EF序列
<213>DNA
<220>
<400>
ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTAAGAA AGTTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACACTGAA TAAGTGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAACCCTC AATTTGCGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTCTGTG CAAAGCAAAATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGGCGAT GACGCAGGCACCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCTACTA TCAGGCGTGAAGGTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGGAAAC CACTCAGGGAGAGAAAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACATCACC AGTGATGAAATTAGGAAAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTGATTA CTGTTCTTCCCGTGCCGCCGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTGGAGG AATGCGTGGGGAGGACGATTTGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAGTGTA CGCGATGCCGAGCAAAACGGATCTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTGCAGT TTCAT。
<210>4
<211>773
<212>粗柄羊肚菌OL-Y-B的RPB1序列
<213>DNA
<220>
<400>
ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTAAGAAAG TTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACACTGAATAAG TGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAACCCTCAATTTG CGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTCTGTGCAAAGCAA AATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGGCGATGACGCAGGCA CCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCTACTATCAGGCGTGAAG GTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGGAAACCACTCAGGGAGAGA AAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACATCACCAGTGATGAAATTAGGA AAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTGATTACTGTTCTTCCCGTGCCGC CGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTGGAGGAATGCGTGGGGAGGACGATT TGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAGTGTACGCGATGCCGAGCAAAACGGAT CTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTGCAGTTTCAT。
<210>5
<211>696
<212>粗柄羊肚菌OL-Y-B的RPB2序列
<213>DNA
<220>
<400>
AAGGGAATGATACTGGCGCAGATACCCAAGATCATACTGGGGTGGATCTCACAATGTGTCCATGTG TGCGCAGTGTGGGGGTTGATCGGCGCCTTGACACGCTTTGCGCTGAACTCATCTTTGGGCTCCTCG TAACCCAGCTGCATCTGCCTACTCATATCCAGATCCTCGGGCGTCATGACGATCATGACCGTCTCC TCTTCCTCTGCATCCAGGTACTCCACGGTACCACTAAACAGAAGACCTTCCCACCCGAACCGGTCC TCCATCGGCAGCGCATGATCGTCTTCCAACTTCTCGATATGCTCCTTCTGGAGCACCAGCTGTCCT CGCCTCTCACTTCGCGGGTCATTGTCAATGACGAACAACGGCCTACATACACGCCCAGCATCTGTA AAAATCTTGAACTCCCTGTCACGAATATCCCTAATGATGGAAACTTCATGGTTGACAACACCCTGC CTTCTTACGTCTTGGACAACCTTGTGCAACATGAGCGGGTCACGATGCGTTCCGACCCAGACACCA TTGACAAATACCTTTGTCGCATTCGGCGATCTCTGCGGCTCATACTCCTCCAGAAGCTCCATATCA TTCGAAGATAAAAAGTCCACGAGCGGGTATCCGGGCGTTCCGATTGTGATGTAGGACATGAGTGAC AAGTTCTTAACGAGACCACACGCCTGTCCCTCGGGC。
核苷酸序列表
<110>西北农林科技大学
<120>一种羊肚菌菌株及其应用
<160>
<210>1
<211>1094
<212> 粗柄羊肚菌OL-Y-B的ITS-nrDNA序列
<213>DNA
<220>
<400> GAGAGAGAGTCAGCTACGATGGGGCGCGCCGGCAAACCAGACCGCTGGCACACTGGTGGCGGGTACTGTCACCCGCGCCGTTTGTCGCAACGTCCAGGGCGAGAGCCTTAGCCGGGTGTGGCCCCATCCGTTAAGCTAGTCAATTCCATGCTGCCTGCTGCGCTCCCTCCCCACGCCCTGGGCAGCAGCTCCCGCAATCGGGGGTGGAAGCCGGATCAAGGAATCAATGCTCTGCATCCATTCAGTCAATTGAATGTGGTGCAGTTTAGGTTATTTCTGCTGAAGGGGGATCCCCCGGCAGAGGTGGACGGCACATGCAGGCTCCGAACGACGGAAGCCAACCATGATGCTGAGTCCCCTGGGGCTAGGGGTGGTAAAAGCCCCTAGCCACTCCCCACCGACGGCGTTGACCCCATCGTTCTGGCACCCTCTCGCCACTGTCCCAACCAAAACCCTCTGTGTACCCTTCCCTGTTGCTTCCCCCGGGAAACTGGCTCCGGCCAGCCGGGGGGAGAAACCAAGCAAAAACCCTTTTCGCAAAACAGACGTCTGAACGCAAAAAACAAAACAAAAAAAAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCCACATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGTATTCCGGAGGGCATGCCTGTTCGAGCGTCATAAATACCGCTCCCCCTCGGATTGCTTGCGGTCCCTGGGGGGTTCTGGCAATGTGGATTCCCCGTGCTTTGAGGGCATGCGAACGGGCTCCCAGTGCTGAAAGACATAATGTTCCCAGCCGAAACCGGTGATTAATTTCATCGGCAGGATTCGTGGCAGGCAGACTGAGGGCGTCAACCGTGGAGTCATGAGGGTAGAAACCTCCCCCTTTGCAAGTAACATTGCTCTGGCAGTTAGATCTGCAGGCCTGCCGGTCTGGGGATGGACCTCCCACTCGCAGGCGTCACGGCCACGATAGCGGGCGTTAAACGGAATCCGATCCGCCCTCACGGGTTGGTTGAAGATCCTTGGGGGCTAG
<210>2
<211>844
<212> 粗柄羊肚菌OL-Y-B的LSU-nrDNA序列
<213>DNA
<220>
<400> CCATGCCCATATTTGACGATCGATTTGCACGTCAGAACCGCTGCGAGCCTCCACCAGAGTTTCCTCTGGCTTCACCCTATACAGGCATAGTTCACCTTCTTTCGGGTCCGGCCCCGTATGCTCTTACTCAAATCCATCCGAGAACATCAGGATCGGTCGGTGATGCGCCGAAGCTCTCACCTACGTTCACTTTCATTACGCGTAGGGGTTTGACACCCGAACACTCGCATACGAAGACGACTCCTTGGTCCGTGTTTCAAGACGGGTCATTGATGACCATTACGCCAGCATCCGTGCCGAAGCGCGTTCCTCAGTCCGCCCCAGGGCATTACACGACGGGCTATAACACTCCCCGAAGGGAGCCACATTCCCGCCGCCTTTATCCCCCGGGGCAAACTGATGCTGGCCTGGACTGGCAGAGTGCACCGGCGAGAACGCCGGATGTTCAACCAAGCCCAAGTCTGGTCATGAATGCTTCCCTTTCAACAATTTCACGTACTTTTTAACCCTCTTTTCAAAGTGCTTTTCATCTTTCGATCACTCTACTTGTGCGCTATCGGTCTCTGGCCGGTATTTAGCTTTAGAAGACGTACACCTCCCATTTAGAGCAGCATTCCCAAACTACTCGACTCGTCGAAGGAGCTTTACAGAGGCTTGGTGGTCCTACCGTACGGGGCTCTCACCCTCTATGGCGTCCCGTTCCAGGGAACTCGGAAGGCACCTCGCCAAAAGCATCCTCTACAAATTACAACTCGGACCTTGCGGCCAGATTTCAAATTTGAGCTGTTGCCGCTTCACTCGCCGTTACTGAGGCAATCCCTGTTGGTTTCTTTTCCTCCGCTTA
<210>3
<211>773
<212> 粗柄羊肚菌OL-Y-B的EF序列
<213>DNA
<220>
<400> ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTAAGAAAGTTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACACTGAATAAGTGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAACCCTCAATTTGCGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTCTGTGCAAAGCAAAATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGGCGATGACGCAGGCACCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCTACTATCAGGCGTGAAGGTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGGAAACCACTCAGGGAGAGAAAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACATCACCAGTGATGAAATTAGGAAAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTGATTACTGTTCTTCCCGTGCCGCCGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTGGAGGAATGCGTGGGGAGGACGATTTGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAGTGTACGCGATGCCGAGCAAAACGGATCTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTGCAGTTTCAT
<210>4
<211>773
<212> 粗柄羊肚菌OL-Y-B的RPB1序列
<213>DNA
<220>
<400> ATCTATTAGAACATGGGGCTTCATTTCGCTAACTCTCCCTAGGGTTTCTTAATAAGATTAAGAAAGTTCTTGAGACAGTTTGCTACAACTGTTCAAAGATCAAGTTAGATGAAGTGAGACACACTGAATAAGTGGCGTGACGTCTTGACTTTCGTTTGGCTAACAGAAATCTACCTCTTCAGTCAAACCCTCAATTTGCGGACGCTTGTAAGATTCGAGAACCGAAAGCCCGTTTTAACGCTGTCTGGCGTCTGTGCAAAGCAAAATCCACTTGTGACTCTGACAACACCGCTGGCGACGAAGAAAACAACTTCGGCGATGACGCAGGCACCAACCCAGAAAAGAAGAAGTCCCACGGGGGTTGTGGTAATAAGCAACCTACTATCAGGCGTGAAGGTCTCCGCCTCATCGGCACTTGGAAACCCGACAAGGATGCACAGGAGGAAACCACTCAGGGAGAGAAAAAGCCAATTCTCCCCTCAGAGGTTCTCAGCATTTTCAAGCACATCACCAGTGATGAAATTAGGAAAATGGGGCTAAGTGAGGATTATGCTCGGCCGGAGTGGATGGTGATTACTGTTCTTCCCGTGCCGCCGCCACCGGTTAGGCCCAGTATTTGTGTGGATGGTATGGGTGGAGGAATGCGTGGGGAGGACGATTTGACATACAAGTTGGCCGAGATCATCCGAGCAAATGCTAGTGTACGCGATGCCGAGCAAAACGGATCTCCAGCTCACGTTGTCAACGAATTTGAGGCCCTCTTGCAGTTTCAT
<210>5
<211>696
<212> 粗柄羊肚菌OL-Y-B的RPB2序列
<213>DNA
<220>
<400> AAGGGAATGATACTGGCGCAGATACCCAAGATCATACTGGGGTGGATCTCACAATGTGTCCATGTGTGCGCAGTGTGGGGGTTGATCGGCGCCTTGACACGCTTTGCGCTGAACTCATCTTTGGGCTCCTCGTAACCCAGCTGCATCTGCCTACTCATATCCAGATCCTCGGGCGTCATGACGATCATGACCGTCTCCTCTTCCTCTGCATCCAGGTACTCCACGGTACCACTAAACAGAAGACCTTCCCACCCGAACCGGTCCTCCATCGGCAGCGCATGATCGTCTTCCAACTTCTCGATATGCTCCTTCTGGAGCACCAGCTGTCCTCGCCTCTCACTTCGCGGGTCATTGTCAATGACGAACAACGGCCTACATACACGCCCAGCATCTGTAAAAATCTTGAACTCCCTGTCACGAATATCCCTAATGATGGAAACTTCATGGTTGACAACACCCTGCCTTCTTACGTCTTGGACAACCTTGTGCAACATGAGCGGGTCACGATGCGTTCCGACCCAGACACCATTGACAAATACCTTTGTCGCATTCGGCGATCTCTGCGGCTCATACTCCTCCAGAAGCTCCATATCATTCGAAGATAAAAAGTCCACGAGCGGGTATCCGGGCGTTCCGATTGTGATGTAGGACATGAGTGACAAGTTCTTAACGAGACCACACGCCTGTCCCTCGGGC
Claims (4)
1.一种羊肚菌菌株,该羊肚菌菌株命名为粗柄羊肚菌OL-Y-B(Morchella crassepsis strain OL-Y-B),在中国微生物菌种保藏管理委员会普通微生物中心的保藏号为CGMCCNo.18115。
2.如权利要求1所述的羊肚菌菌株,其特征在于,所述的粗柄羊肚菌OL-Y-B的培养条件是:在黑暗条件下,采用pH 7.0-8.5的PDA培养基,培养温度为25℃-30℃。
3.权利要求1所述的羊肚菌菌株用于促进糜子、玉米、小麦或荞麦共生的应用。
4.如权利要求3所述的应用,其特征在于,所述的粗柄羊肚菌OL-Y-B经发酵培养后的发酵液菌体和PDA平板培养产生的菌核和菌丝体,用于替代化肥制备促生长的生物制剂,所述的发酵培养条件为:PD液体培养基,pH值 7.0-8.0, 转速80-150rpm,25-30℃;PDA固体培养基,pH值 7.0-8.0,黑暗培养2周。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1793316A (zh) * | 2005-12-08 | 2006-06-28 | 云南菌苑科技有限公司 | 黑脉羊肚菌菌株的筛选及菌种制备方法 |
CN102174414A (zh) * | 2010-12-28 | 2011-09-07 | 青海大学 | 一种新的肋脉羊肚菌m8-13液体发酵物在保健品及医药开发中的应用 |
CN103907481A (zh) * | 2014-04-20 | 2014-07-09 | 丽江中源绿色食品有限公司 | 一种羊肚菌优质菌种的生产工艺 |
KR20150108064A (ko) * | 2014-03-17 | 2015-09-25 | 주식회사 앤코스메슈 | 모렐버섯의 생산을 위한 배지 조성물 |
CN108410854A (zh) * | 2018-03-21 | 2018-08-17 | 福建师范大学 | 一种制备蛹虫草菌与羊肚菌融合菌株的方法 |
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1793316A (zh) * | 2005-12-08 | 2006-06-28 | 云南菌苑科技有限公司 | 黑脉羊肚菌菌株的筛选及菌种制备方法 |
CN102174414A (zh) * | 2010-12-28 | 2011-09-07 | 青海大学 | 一种新的肋脉羊肚菌m8-13液体发酵物在保健品及医药开发中的应用 |
KR20150108064A (ko) * | 2014-03-17 | 2015-09-25 | 주식회사 앤코스메슈 | 모렐버섯의 생산을 위한 배지 조성물 |
CN103907481A (zh) * | 2014-04-20 | 2014-07-09 | 丽江中源绿色食品有限公司 | 一种羊肚菌优质菌种的生产工艺 |
CN108410854A (zh) * | 2018-03-21 | 2018-08-17 | 福建师范大学 | 一种制备蛹虫草菌与羊肚菌融合菌株的方法 |
Non-Patent Citations (1)
Title |
---|
北秦岭羊肚菌的***学分析及其对玉米和小麦的促生作用;彭博;《中国优秀硕士学位论文全文数据库》;20190815(第8期);摘要、第三-四章、附图18、21、25-26 * |
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