CN101522023A - 改善植物健康的方法和组合物 - Google Patents
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Abstract
本发明提供了改善植物健康的方法和组合物。具体地,向植物施用麦草畏或DMO的另一种底物或其代谢物(包括DSCA)提供了对非生物性应激或生物性应激如氧化应激(包括施用除草剂)和植物病害的耐受性或抵抗,并提高了农作物产量。这样的施用可以与如草甘膦的另一种除草剂的施用联合。
Description
发明背景
本申请要求2006年10月16日提交的美国临时申请第60/852,308号的优先权,本文引入其完整公开内容作为参考。
技术领域
本发明一般地涉及农业领域。更具体地说,本发明涉及通过向植物施用麦草畏除草剂和/或其代谢物或类似物而改善植物健康的方法。
相关现有技术的说明
麦草畏(3,6-二氯-邻-茴香酸或3,6-二氯-2-甲氧基苯甲酸;图7),是如(BASF)、(BASF)和(Syngenta)的除草剂中的活性成分,是一种强效除草剂。尽管其活性的确切机制还不清楚,其似乎是作为植物生长调节剂发挥作用(例如,Grossmann2000)。其施用导致不受控制的生长、叶子卷曲和扭转、叶绿体损伤和直接的植物毒性效应等。据信其中的某些效应由乙烯合成导致,乙烯合成引发了另一种植物激素脱落酸的生物合成的增加。因此,植物激素水平的失衡似乎是毒性效应的原因。
已发现麦草畏单氧合酶(DMO)在细菌中通过将麦草畏降解成3,6-二氯水杨酸(DCSA,3,6-DCSA,图7)而提供对麦草畏的耐受性(例如,Herman等人,2005;美国专利公开20060168700;U.S.7,022,896)。随后应用DMO基因给大豆和其它植物提供对麦草畏的耐受性(例如,Weeks等人,2006)。这些新的耐受麦草畏的农作物允许对以前对于任何麦草畏暴露极其敏感的农作物、尤其是如棉花、油菜和大豆的双子叶植物施用麦草畏。
美国专利7,230,163描述了通过向植物施用合成的生长素来提高农作物产量的方法。但是,没有提到植物的健康,例如,对于生物性或非生物性应激的抗性,也没有提到施用的生长素的代谢物的效应。
由于植物中的代谢一般是不可预测的,即,人们不能从麦草畏的以前的传统使用(例如,对于不表达DMO基因的玉米使用)预测在新的表达DMO的麦草畏耐受农作物中使用麦草畏会产生什么样的代谢物,或者这样的代谢物在植物中产生什么效应。本发明人发现,在这些表达DMO的农作物中,DMO对麦草畏解毒,并产生包括DSCA的麦草畏的代谢物。出乎意料地,本发明人还发现,这种DSCA代谢物给予植物有益的健康效应。因此,本发明尤其涉及使用包括DSCA的麦草畏代谢物改善植物健康的方法和由此产生的植物、种子和农作物。
这些在植物健康方面的改善提供了植物对生物性应激(例如,昆虫、真菌、病毒、线虫和其它病原体)和非生物性应激(例如,干旱、寒冷、臭氧和土壤养分缺乏)的抗性提高,导致产量增加和农作物的质量提高,所有这一切都是对于农业的巨大的益处。
附图说明
图1是在用麦草畏处理DMO植物后各个时间点的PR2的诱导。
图2是在用各种施用量的麦草畏和DSCA处理两种独立的大豆转基因系24小时后,PR2的诱导。
图3是在存在或不存在3,6-DCSA的情况下,用终极腐霉(Pythium ultimum)处理棉花植物的效应。
图4是在存在或不存在3,6-DCSA的情况下,用野油菜黄单胞菌锦葵致病变种(Xanthomonas campestris pv.malvacearum)处理棉花植物的效应。
图5显示在大豆中提供DMO基因对产量和其它农学性状没有不利影响。
图6是麦草畏喷施对于产量的影响。
图7是麦草畏和3,6-DCSA的化学结构。
发明概述
一方面,本发明提供了改善植物健康的方法,包括向植物提供一定量的麦草畏或DMO所介导代谢的产物或其类似物,与没有向其提供麦草畏或DMO所介导代谢的产物或其类似物的相同基因型的植物相比,所述量改善植物的健康。在一种实施方式中,本发明提供了改善植物健康的方法,包括将植物接触麦草畏或其DMO介导的代谢产物,以向该植物提供一定量的DMO介导的麦草畏代谢的产物,与没有向其提供麦草畏或其DMO介导的代谢产物的相同基因型的植物相比,所述量改善植物的健康,其中,接触麦草畏的植物包含DMO。
在一种实施方式中,植物在农作物生产田地中。该方法进一步包括在向植物提供麦草畏或其DMO介导的代谢产物或其类似物之前、同时或之后,植物遭受生物性或非生物性应激。该方法也可进一步包括在向植物提供麦草畏或其DMO介导的代谢产物或其类似物之前,确认植物需要改善植物健康的步骤。
在一种实施方式中,确认植物需要改善植物健康包括确认植物显示至少指示生物性或非生物性应激的第一症状。在具体的实施方式中,所述症状选自矮化、光合作用功能丧失、脂质过氧化作用、活性氧物质的积聚、自由基含量增加和组织坏死(也称为“超敏应答”)。在一种特别的实施方式中,植物显示矮化的症状。确认为需要改善健康的植物可以是营养生长期和对病害或杂草生长敏感的不成熟的植物。在特定的实施方式中,所述植物是处于VE到V3生长阶段的大豆植物、第一棉蕾形成之前的棉花植物或处于VE到VT生长阶段之前或之间的玉米植物。
根据本发明改善其健康的植物可能处于非生物性应激的危险或在非生物性应激之下。非生物性应激的例子包括渗透应激、热或寒冷暴露、氧化应激和营养缺乏。在一种具体的实施方式中,植物处于如干旱应激的渗透应激的危险或在渗透应激之下。在另一种实施方式中,植物处于例如由于施用除草剂或由于存在能够损伤植物的臭氧水平而造成的氧化应激的危险或在该氧化应激之下。
也可以限定这样的植物处于生物性应激的危险或在生物性应激之下。生物性应激的例子包括如大豆锈病的真菌病、病毒病、细菌病、昆虫侵扰、线虫侵扰和杂草侵扰。在一种实施方式中,植物处于真菌病应激的危险或在真菌病的应激之下。在一种具体的实施方式中,植物处于大豆锈病的危险或在大豆锈病之下。在另一种实施方式中,植物处于杂草侵扰的危险或在杂草侵扰之下。杂草侵扰构成对于农作物、尤其是对于如处于至多V3-V4生长阶段的大豆植物的幼小植物的应激。
在本发明的一种方法中,可以增加植物对于氧化应激的耐受性。本发明的一种方法也可以包括向植物群提供麦草畏或DMO所介导代谢的产物或其类似物,以改善植物的健康,例如,通过将麦草畏代谢为DCSA。如果施用麦草畏,这可能为不除草的施用量,但由于麦草畏代谢为DCSA或麦草畏或DCSA的植物代谢的其它产物,仍然导致对植物健康的益处。
在本发明的一种方法中,可以限定DMO介导的代谢的产物为3,6-DCSA、3,5-DCSA或3-CSA,或3,6-DCSA、3,5-DCSA和3-CSA的代谢物中的一种或多种。麦草畏、应用的DMO介导的代谢产物或代谢物,可以是除草性的。植物可以包含编码DMO的转基因。应用的产物或应用的产物的代谢物,也可以是非除草性的。在一种实施方式中,向田地使用的施用量(application rate)是大约0.0025磅/英亩(lb/a)到大约9lb/a麦草畏,包括大约0.25lb/a到大约1.5lb/a,包括,例如大约0.25lb/a到大约1lb/a、大约0.5lb/a到大约1.5lb/a和大约0.5lb/a到大约1lb/a,以及更低或更高的量及它们之间的全部范围,如大约0.005lb/a、大约0.01lb/a、大约0.025lb/a、大约0.05lb/a、大约0.15lb/a、大约0.175lb/a、大约2lb/a、大约4lb/a、大约7lb/a和大约12lb/a。在另一种实施方式中,使用大约0.0025lb/a到大约12lb/a DCSA,包括,例如大约0.25lb/a到大约4lb/a、大约0.5lb/a到大约6lb/a和大约4lb/a到大约12lb/a,以及更低或更高的量及它们之间的全部范围,包括大约0.005lb/a、大约0.01lb/a、大约0.025lb/a、大约0.05lb/a、大约0.15lb/a、大约0.175lb/a、大约0.25lb/a、大约0.5lb/a、大约1lb/a、大约3lb/a、大约5lb/a、大约8lb/a、大约12lb/a、大约15lb/a和大约20lb/a。在本发明的某些实施方式中,可以以非除草的施用量反复施用麦草畏或DMO介导的代谢产物。
在具体的实施方式中,通过向植物施用麦草畏而向植物(包括其任意部分)提供DCSA,并允许植物代谢以产生DSCA。以这种方式,通过直接施用麦草畏能够获得健康益处。可选择地,可以直接施用DSCA或一种不同的麦草畏的代谢产物。
用于本发明的实施方式的植物可以包含编码DMO的转基因。植物可以是双子叶植物。双子叶植物的例子包括苜蓿、豆类、甜菜、椰菜、卷心菜、油菜、胡萝卜、花椰菜、芹菜、大白菜、棉花、黄瓜、茄子、亚麻、洋姜、莴苣、羽扇豆、甜瓜、豌豆、胡椒、花生、马铃薯、南瓜、萝卜、油菜籽、菠菜、大豆、倭瓜、糖用甜菜、向日葵、番茄和西瓜。在某些实施方式中,植物选自棉花、油菜或如大豆的豆科植物、苜蓿。在一种具体的实施方式中,植物是大豆植物(大豆(Glycine max))。在另一种实施方式中,植物是棉花植物(棉属的种(Gossypium sp),如陆地棉(G.hirsutum))。
在某些实施方式中,植物可以是单子叶植物。单子叶植物的例子包括大麦、玉米、韭菜、洋葱、水稻、高梁、甜玉米、小麦、黑麦、小米、甘蔗、燕麦、黑小麦、柳枝稷和草坪草。在某些实施方式中,植物可以是谷物(禾本科(Gramineae)),如玉米(Zea mays)。在一种具体的实施方式中,植物是玉米。
可以进一步限定植物耐受选自草甘膦(glyphosate)、草丁膦(glufosinate)、2,4-D、异噁唑草酮(isoxaflutole)、麦草畏和磺酰脲的除草剂,包括其任意组合,并可以用任意这样的除草剂来处理植物。在某些实施方式中,植物耐受草甘膦和磺酰脲,或耐受草丁膦和磺酰脲。在其它的实施方式中,植物耐受草甘膦和麦草畏。在一种具体的实施方式中,植物由于存在对麦草畏解毒的转基因而耐受麦草畏。在另一种具体的实施方式中,植物不包括对麦草畏解毒的转基因。
在另一方面,本发明提供了提高植物产量的方法,包括将植物与一定量的麦草畏或DMO介导的麦草畏代谢的产物接触,相对于在同样的条件下生长但不接触麦草畏或DMO介导的麦草畏代谢的产物的相同基因型的植物,所述量有效提高植物的产量。植物可以在农作物生产田地中。该方法可进一步包括将植物群体接触麦草畏或其DMO介导的代谢产物。DMO介导的代谢产物可以为3,6-DCSA、3,5-DCSA或3-CSA,或3,6-DCSA、3,5-DCSA或3-CSA的代谢物,如DCGA(5-OH DCSA,DC-2,5-二羟基苯甲酸)。
在本方法的具体的实施方式中,植物可以包含编码DMO的转基因。植物可以是,例如,如上所列出的双子叶或单子叶植物。可以进一步限定植物耐受选自草甘膦、草丁膦、2,4-D、硝草酮(mesotrione)、噻草定、异噁唑草酮、溴苯腈、莠去净(atrazine)、精吡氟禾草灵(Fluazifop-P)和磺酰脲类/咪唑啉酮类的除草剂,包括其任意组合,并可以用任意这样的除草剂来处理植物。
本发明的另一种实施方式包括改善种子健康的方法,包括将种子与一定量的麦草畏或其DMO介导的代谢产物接触,与不接触麦草畏或其DMO介导的代谢产物的相同基因型的种子相比,所述量改善种子的健康。
在另一方面,本发明提供生产3,6-DCSA的方法,包括将农作物生产田地中的植物群体与麦草畏接触,其中,所述植物包括编码DMO的转基因。
发明详述
根据本发明,提供了改善植物健康的方法,包括增强植物的抗病力,给予植物提高的对氧化应激的耐受性,和/或提高植物的产量。由于麦草畏对于许多植物种类通常是高毒性的,因此提供麦草畏或其代谢物如3,6-DCSA或DCGA(5-OH DCSA,DC-2,5-二羟基苯甲酸)而导致的植物健康和其它益处是尤其令人惊奇的。“麦草畏”指3,6-二氯-邻-茴香酸或3,6-二氯-2-甲氧基苯甲酸(图7)及其酸和盐。其盐包括异丙胺盐、二甘醇胺盐(diglycoamine)、二甲胺盐、钾盐和钠盐。麦草畏的商业制剂的例子包括,但不限于,(作为DMA盐)、(BASF,作为DGA盐)、VEL-58-CS-11TM和(BASF,作为DGA盐)。
因而,在一方面,本发明涉及以下令人惊奇的发现,用麦草畏处理包括麦草畏单氧合酶(DMO)转基因的抗麦草畏植物改善了该植物的健康。这样的健康益处可以包括,例如,植物对生物性或非生物性应激的抗性。在具体的实施方式中,麦草畏、3,6-DCSA(见,例如,图7)及其类似物可以用来获得一种或多种选自抗病力、氧化应激抗性及产量的健康益处。在进一步的实施方式中,这样的处理可以诱导一般称为“系统获得性抗性”(“SAR”)的植物应答。
麦草畏、3,6-DCSA、DCGA和DMO介导的麦草畏代谢的其它产物及DMO的底物因而可以诱导植物健康的改善。“DMO介导的麦草畏代谢的产物”可以包括3,6-DCSA、3,6-DCSA的代谢产物或其类似物。响应施用麦草畏或DCSA产生的蛋白质可能具有直接的抗微生物活性(例如,与发病机理相关的蛋白质,如几丁质酶),或可能具有加强植物健康改善、抗病力、氧化应激抗性和产量提高中的一种或多种的另一种功能。因此,本发明的一方面是生产3,6-DCSA或DCGA的方法,包括将在农作物生产田地中的植物群体接触麦草畏。所述植物可以是包含DMO转基因的转基因植物。
在本发明的某些方面,可以将植物接触DMO介导的植物内麦草畏代谢的产物,如3,6-DCSA或DCGA,使得与没有接触麦草畏或DMO介导的麦草畏代谢的产物而其它方面相同的植物相比,该植物显示提高的应激抗性。
在一种实施方式中,可以限定植物缺乏DMO转基因,并将植物接触DMO介导的麦草畏代谢的非除草性产物,如3,6-DCSA。现发现,例如,接触3,6-DCSA的棉花植物在接种植物致病性卵菌—终极腐霉(Pythium ultimum)后存活下来,而在相同的条件下终极腐霉杀死了在相同条件下生长的其它方面相同但未接触3,6-DCSA的植物。
另一方面,植物可以接触麦草畏、3,6-DCSA、DCGA和/或DMO介导的植物内代谢的其它产物,这产生提高的氧化应激耐受性。这样的应激可以是环境的,或,例如,由除草剂、病原体或如臭氧的其它物质的存在而引起。在一种实施方式中,提高氧化应激耐受性,使得植物的光合作用活性没有由于存在氧化应激而减少,或较少受到影响。可以通过本领域熟知的方法来测定光合作用活性,例如,通过测量经过光合系统I和/或II的电子转移(例如,Peterson和Arntzen,1982;Allen和Holmes,1986)。在另一种实施方式中,如果植物已经接触麦草畏、3,6-DCSA或DMO介导的麦草畏代谢的另一种产物,则植物的生长、发育、开花或结果不会由于存在氧化应激而受到有害地影响,或较少受到影响。在另一实施方式中,氧化应激的坏死效应减少。
在本发明的具体的实施方式中,可以限定根据本发明处理的植物在农作物生产田地中生长。“农作物生产田地”是指一种生长环境,其中一般为了包括种子生产的生产目的而在田地中种植农作物植物,而非在实验室温室中。在本发明的另外的实施方式中,可以限定植物群体在农作物生产田地中生长并按照本发明处理。按照本发明处理的植物可以是经受营养生长和对病害或杂草压力敏感的不成熟的植物,如处于VE到V3-V4生长阶段的大豆植物。所述植物也可以处于较晚的生长阶段。
3,6-DCSA的化学结构如下式(I):
例如,可以限定3,6-DCSA的类似物为取代的苯甲酸及其生物学上可接受的盐,其中苯甲酸上的取代可以包括在3-、4-、5-和/或6-位的单-、二-、三-或四-取代。例如,取代基可以选自:1-4个碳的低级烷基;卤素氟、氯、溴或碘;氨基,其中氮原子可以携带0、1或2个相同或不同的各自有1-4个碳的低级烷基;硝基;甲酰基;乙酰基;羟甲基;甲氧羰基;羟基;烷硫基、烷基亚磺酰基或烷基磺酰基,其中烷基由1-4个碳组成,或单-、二-或三-氟甲基。生物学上可接受的盐包括那些普通的碱金属钠盐和钾盐、碱土金属镁盐或钙盐、锌盐或铵盐,或简单的烷基铵阳离子,如单-、二-、三-或四甲基铵阳离子。
DMO介导的代谢产物可以缀合成糖苷,糖苷可以水解为苷元以调节(例如,延长)健康益处。在某些实施方式中,预期与如葡萄糖、半乳糖或甘露糖等糖缀合。在其它实施方式中,预期与氨基酸(其非限制性的例子包括丙氨酸、亮氨酸、天冬氨酸或谷氨酸)缀合。
在另一方面,与没有接触麦草畏或3,6-DCSA或DMO介导的代谢的其它产物但在相同条件下生长的相同基因型的植物的产量相比,麦草畏或3,6-DCSA或DMO介导的代谢的其它产物的应用或存在使植物的产量提高。在某些实施方式中,植物可以是大豆、棉花、油菜籽或玉米植物等。
在另外一方面,植物可以与非除草性的前体分子接触,该前体分子在植物内转化为SAR诱导性的代谢物。在某些实施方式中,植物可以包含DMO转基因,且产生的编码的DMO可以将前体分子转化为SAR诱导性的代谢物。在具体的实施方式中,可以向植物施用2-甲氧基,3,5-二氯-苯甲酸或2-甲氧基,3-氯-苯甲酸,它们分别被DMO转化为3,5-DCSA或3-CSA,以获得植物健康改善、植物抗病力增加、植物的氧化应激耐受性提高和/或植物农业产量提高中的任一种或多种。
在一种实施方式中,本发明的方法可以用于双子叶农作物植物。这样的双子叶植物的非限制的例子包括苜蓿、豆类、甜菜、椰菜、卷心菜、油菜、胡萝卜、花椰菜、芹菜、大白菜、棉花、黄瓜、茄子、莴苣、甜瓜、豌豆、胡椒、花生、马铃薯、南瓜、萝卜、油菜籽、菠菜、大豆、倭瓜、糖用甜菜、向日葵、番茄和西瓜。在某些实施方式中,双子叶植物是大豆、棉花或油菜籽。在其它实施方式中,本发明的方法可以用于单子叶农作物植物,包括但不限于,大麦、玉米、韭菜、洋葱、水稻、高粱、甜玉米、小麦、黑麦、小米、甘蔗、燕麦、黑小麦、柳枝稷和草坪草。
生物性和非生物性的作物应激条件可以包括,例如,干旱、缺乏光照、真菌病、病毒病、细菌病、昆虫侵扰、线虫侵扰、杂草侵扰、低温暴露、热暴露、渗透应激、氧化应激、氮养分可用性减少、磷养分可用性减少和高植物密度。这样的条件可能对植物不利,不利地影响植物代谢、生长和/或发育。
本发明的方法在控制、预防或治疗多种植物病害方面得到应用。本发明的方法包括对于植物病原体感染的预防性抑制和治疗性处理。植物病原体可以根据其与植物宿主有关的生命周期进行分类,这些分类包括,专性寄生物、兼性寄生物和兼性腐生物。专性寄生物只能够通过从活的植物细胞获得营养而生存并繁殖,并直接与这些细胞接触,植物的专性真菌寄生物的例子包括,但不限于,锈菌目(Uredinales)(锈病)、黑粉菌目(Ustilaginales)(黑粉病和腥黑穗病)、白粉菌目(Erysiphales)(白粉病)和卵菌纲(水霉和霜霉)的成员。兼性寄生物是一般作为在其它生物体的产物或死的生物体上的腐生物生存,但当条件有利时可变为寄生物的生物体。兼性腐生物是一般作为植物的寄生物生存,但当易感植物宿主不可用时作为腐生物生存的生物体。本发明的方法可用于控制、预防或治疗包括专性寄生物、兼性寄生物和兼性腐生物的多种植物病原体的感染,这些植物病原体包括,但不限于:
子囊菌纲(Ascomycete)真菌,如黑星菌属(Venturia)、叉丝单囊壳属(Podosphaera)、白粉菌属(Erysiphe)、丛梗孢属(Monilinia)、球腔菌属(Mycosphaerella)和钩丝壳属(Uncinula);担子菌纲(Basidiomycete)真菌,如驼孢锈菌属(Hemileia)、丝核菌属(Rhizoctonia)和柄锈菌属(Puccinia);半知菌类(Fungi imperfecti),如葡萄孢属(Botrytis)、长蠕孢属(Helminthosporium)、喙孢属(Rhynchosporiurn)、镰孢属(Fusarium)(如,串珠镰孢(F.monoliforme))、壳针孢属(Septoria)、尾孢属(Cercospora)、链格孢属(Alternaria)、梨孢属(Pyricularia)、假尾孢属(Pseudocercosporella)(如,P.herpotrichoides)和轮枝孢属(Verticillium);卵菌纲真菌,如疫霉属(Phytophthora)(如,寄生疫霉(P.parasitica)、苜蓿疫霉(P.medicaginis)、大雄疫霉(P.megasperma))、霜霉属(Peronospora)(如,烟草霜霉(P.tabacina))、盘梗霉属(Bremia)、腐霉属(Pythium)和单轴霉属(Plasmopara);以及其它真菌,如指疫霉(Scleropthora macrospora)、玉米褐条指疫霉(Sclerophthorarayissiae)、禾生指梗霉(Sclerospora graminicola)、Peronosclerosporasorghi、Peronosclerospora philippinensis、Peronosclerospora sacchari和Peronosclerospora maydis、玉米壳锈菌(Physopella zeae)、Cercospora zeae-maydis、禾生刺盘孢(Colletotrichum graminicola)、玉蜀黍赤霉(Gibberella zeae)、Exserohilum turcicum、玉蜀黍球梗孢(Kabatiella zeae)和Bipolaris maydis可以导致通过本发明的方法可控制、预防或治疗的病害。尤其优选的病原体包括,但不限于:柄锈菌属、丝核菌属、GGT(禾顶囊壳小麦变种(Gaeumannomycesgraminis var.tritici))、条锈病、层锈菌属的种(Phakopsora sp.)包括豆薯层锈菌(P.pachyrhizi)(导致亚洲大豆锈病)、镰孢属的种、如大丽花轮枝孢(V.dahliae)的轮枝孢属的种、Cercosporazeae-maydis(导致灰叶斑病)、疫霉属的种和玉米锈病。因此,可控制、预防或治疗的病害包括,例如,苜蓿植物的病害,如根腐病(苜蓿疫霉(Phytophora medicaginis)、大雄疫霉);水稻植物的病害,如稻瘟病(Pyricularia oryzae)、长蠕孢属叶斑病(稻长蠕孢(Helminthosporium oryzae)、宫部旋孢腔菌(Cochliobolusmiyabeanus))、赤霉病(腾仓赤霉(Gibberella fujikuroi))、苗绵腐病(米根霉(Rhizopus oryzae))、鞘枯萎病(立枯丝核菌(Rhizoctoniasolani))等;燕麦的病害,如冠锈病(禾冠柄锈菌(Puccinia coronata))等;大麦的疾病,如白粉病(禾白粉菌(Erysiphe graminis))、褐斑病(黑麦喙孢(Rhynchosporium secalis))、斑枯(禾旋孢腔菌(Cochliobolus sativus))、黄斑叶病(禾长蠕孢(Helminthosporiumgramineum)、麦类核腔菌(Pyrenophora gramineum))、网斑病(圆核腔菌(Pyrenophora teres))、腥黑穗病(小麦网腥黑粉菌(Tilletiacaries))、散黑粉病(裸黑粉菌(Ustilago nuda))等;小麦的疾病,如白粉病(禾白粉菌)、麦类颖斑病(颖枯壳小球腔菌(Leptosphaeria nodorum)、颖枯壳针孢(Septoria nodorum))、条锈病(条形柄锈菌(Puccinia striiformis))、麦类雪腐菌核病(肉孢核瑚菌(Typhula incarnata))、眼斑病(Pseudocercosporellaherpotrichoides)、雪霉病(禾生丽赤壳(Calonectria graminicola)、雪腐镰孢(Fusarium nivale))、茎锈病(禾柄锈菌(Pucciniagraminis))、黑雪枯萎病(Typhula ishikariensis)、疮痂病(玉蜀黍赤霉)、叶锈病(隐匿柄锈菌(Puccinia recondita)、Pucciniatriticina)、条纹病(禾长蠕孢)、腥黑穗病(小麦网腥黑粉菌)、斑点叶枯病(小麦壳针孢(Septoria tritici))、散黑粉病(小麦散黑粉菌(Ustilago tritici))等;草坪草的病害,如灰叶斑病(稻梨孢菌(Pyricularia grisea))等;玉米的病害,如玉米锈病、猝倒病(德巴利腐霉(Pythium debaryanum))等;黑麦的病害,如紫色雪霉病(雪腐镰孢)等;棉花的病害,如黄萎病(大丽花轮枝孢)、幼苗病(终极腐霉、棉刺盘孢(Colletotrichum gossypii)、腐霉属的种、立枯丝核菌、根串珠霉属的种(Thielaviopsis sp.))、细菌性黑枯病(野油菜黄单胞菌锦葵致病变种)等;马铃薯的病害,如晚疫(致病疫霉(Phytophthora infestans))等;烟草植物的病害,如霜霉病(烟草霜霉(Peronospora tabacina))、根腐病(烟草寄生疫霉(Phytophthora parasitica var))、壳针孢属枯萎病(烟草尾孢(Cercospora nicotianae))等;甜菜的病害,如叶斑病(菾菜生尾孢(Cercospora beticola))、猝倒病(德巴利腐霉、立枯丝核菌、瓜果腐霉(Pythium aphanidermatum))等;红辣椒的病害如,灰霉(灰葡萄孢(Botrytis cinerea))等,菜豆的病害,如灰霉(灰葡萄孢)、核盘菌属种子腐烂(核盘菌属腐烂;核盘菌(Sclerotiniasclerotiorum))、白绢病(罗耳伏格菌(Corticium rolfsii))等;蚕豆的病害,如白粉病(蓼白粉菌(Erysiphe polygoni)、单丝壳(Sphaerotheca fuliginea)),锈病(蚕豆单胞锈菌(Uromyces fabae)、菜豆单胞锈菌(Uromyces phaseoli))、灰霉病(灰葡萄孢)等;花生的病害,如壳二孢属斑点病(落花生球腔菌(Mycosphaerellaarachidicola))等;卷心菜的病害,如猝倒枯萎病(立枯丝核菌)等;黄瓜的病害,如白粉病(单丝壳)、茎腐(尖镰孢(Fusariumoxysporum))、茎部腐烂病(甜瓜球腔菌(Mycosphaerella melonis))、霜霉病(古巴假霜霉(Pseudoperonospora cubensis))、灰霉病(灰葡萄孢)、核盘菌属种子腐烂(Sclerotial seed rot)(核盘菌)、炭疽病(葫芦科刺盘孢(Colletotrichum lagenarium))、猝倒枯萎病(Damping blight)(尖镰孢、瓜果腐霉、立枯丝核菌)等;小松菜(即,芸苔(Brassica rapa var.))的病害,如链格孢属烟斑(甘蓝链格孢(Alternaria brassicicola))、根肿病(芸苔根肿菌(Plasmodiophora brassicae))等;芹菜的病害,如斑点叶斑病(芹菜小壳针孢(Septoria apii))等;萝卜的病害,如枯黄病(尖镰孢)等,番茄的病害,如镰刀菌萎缩病(尖镰孢)、根腐病(致病疫霉)、菌环叶斑病(Alternaria solani)、灰霉(灰葡萄孢)、叶斑病(辣椒疫霉(Phytophthora capsici))、黑腐病(番茄链格孢(Alternariatomato))等;茄子的病害,如褐腐(辣椒疫霉)、脉管枯萎病原体,例如,黄萎病(黄萎轮枝孢(Verticillium albo-atrum.)、大丽花轮枝孢)等;大白菜的病害,如黑腐病(日本链格孢(Alternaria japonica))、根肿病(芸苔根肿菌)等;甜椒的病害,如根腐病(辣椒疫霉)、灰霉(灰葡萄孢)等;莴苣的病害,如灰霉(灰葡萄孢)等;柑桔类水果的病害,如荚果和茎疫病(柑桔间座壳(Diaporthe citri))等;梨的病害,如疮痂病(梨黑星菌(Venturia nashicola))、黑腐病(菊池链格孢(Alternaria kikuchiana))、日本梨锈病(梨胶锈菌(Gymnosporangium haraeanum))、褐斑病(由黄花菜匍柄霉(Stemphylium vesicarium)导致)等;葡萄的病害,如霜霉病(葡萄生单轴霉(Plasmopara viticola))、灰霉(灰葡萄孢)、痂园孢属疮痂病(痂囊腔菌( ampelina))等;桃的病害,如缩叶病(畸形外囊菌(Taphrina deformans))、疮痂病(嗜果枝孢(Cladosporium carpophilum))、穿孔病(樱桃球腔菌(Mycosphaerellacerasella))等;苹果的病害,如白粉病(白叉丝单囊壳(Podosphaeraleucotricha))、疮痂病(苹果黑星菌(Venturia inaequalis))、灰霉(灰葡萄孢)、黑腐病(Botryosphaeria obtusa)、褐斑病(山田胶锈菌(Gymnosporangium yamadae))、白色根腐病(褐座坚壳(Rosellinia necatrix))、链格孢属叶斑病(苹果链格孢(Alternariamali))等;和其它谷物、水果和蔬菜的病害,如油菜(oil-seed rape)、向日葵、胡萝卜、胡椒、草莓、甜瓜、猕猴桃、洋葱、韭菜、甘薯、无花果、梅花、芦笋、柿子、大豆、赤小豆、西瓜、茼蒿菜、菠菜、茶等。
也包括病毒、细菌和线虫导致的病害,如病毒花叶病(例如,由烟草花叶病毒、大豆花叶病毒、苜蓿花叶病毒或黄瓜花叶病毒引起)、大豆矮缩病毒和豆荚斑点病毒等;由细菌引起的,包括丁香假单胞菌致病变种(Pseudomonas syringae pvs.),如丁香假单胞菌大豆致病变种(P.syringae pv.Glycinea)、晕斑假单胞菌(P.syringaepv.Coronafaciens),野油菜黄单胞菌致病变种(Xanthomonascampestris pvs.),如野油菜黄单胞菌大豆病原变种(X.campestris pv.Glycines)、水稻黄单胞菌(Xanthomonas oryzae)、半透明黄单胞菌(Xanthomonas translucens)、地毯黄单胞菌(Xanthomonasaxonopodis pv.Malvacearum)(棉花细菌性黑枯病);欧文氏菌属的种(Erwinia spp.)和包括泛菌属的种(Pantoea spp.)、和棍状杆菌属的种(Clavibacter spp.)等,以及由如根结线虫(Meloidogyneincognita)、穿刺短体线虫(Pratylenchus penetrans)、剑线虫属的种(Xiphinema sp.)和胞囊线虫属的种(Heterodera sp.)等线虫引起的病害。
在本发明的一种实施方式中,作物的生物性应激是锈菌(担子菌)。一些农业上重要的植物锈病包括,但不限于,由柄锈菌属的种引起的,如由禾冠柄锈菌、禾柄锈菌、条形柄锈菌、高粱柄锈菌(Puccinia sorghi)、多堆柄锈菌(Puccinia polysora)和隐匿柄锈菌(P.recondita)引起的谷类锈病;由胶锈菌属的种(Gymnosporangiumsp.)引起的锈病;由茶藨生柱锈菌(Cronartium ribicola)引起的白洋松疱锈病;由咖啡驼孢锈菌(Hemileia vastatrix)引起的咖啡锈病;和由单胞锈菌属的种(Uromyces sp.)引起的锈病。在一种具体的实施方式中,作物是大豆,作物的生物性应激是由层锈菌属的种引起的大豆锈病。
在本发明的某些方面,可以单独向植物提供或应用麦草畏或3,6-DCSA,或与另一种除草剂或其它活性成分联合应用。可以在应用麦草畏、3,6-DCSA或DMO介导的麦草畏代谢的其它产物之前、同时或之后应用其它除草剂。
“向植物应用”也可以包括向种子应用麦草畏、3,6-DCSA或DMO介导的麦草畏代谢的其它产物。例如,可以根据发芽的百分率、到发芽的时间、对幼苗病害或应激的抗性、幼苗活力,或根据获得的作物的挺立(stand),来确定种子的“健康”。
基于公开的内容,制备用于本发明的这样的组合物对于本领域的技术人员来说是显而易见的。这些组合物除了活性成分外一般还包含如表面活性剂、固体或液体载体、溶剂和粘合剂的组分。可用于向植物施用的表面活性剂的例子包括以下酸的碱金属盐、碱土金属盐或铵盐:芳族磺酸,例如,木素-、苯酚-、萘-和二丁基萘磺酸,和芳基磺酸盐的脂肪酸、烷基醚的脂肪酸、月桂基醚的脂肪酸、脂肪醇硫酸盐的脂肪酸和脂肪醇二醇醚硫酸盐的脂肪酸,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、三丁基苯基聚乙二醇醚、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、月桂醇聚乙二醇醚乙酸酯、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%,更普遍地为大约0.25重量%-0.5重量%。
在本发明的一种实施方式中,可以联合草甘膦提供麦草畏。“联合”意思是麦草畏可以与草甘膦同时或在其之前或之后施用。由于协同作用,可以使用这种实施方式来减少每种除草剂的量以达到与只施用一种除草剂相同的活性程度。例如,本发明可以涉及施用相对于标准生产者标出的施用量低于1x的施用量的草甘膦和/或麦草畏。草甘膦和麦草畏各自的施用量的例子,除了1x的量外,还包括大约0.25x-0.95x的每种除草剂,特别包括大约0.5x、0.6x、0.7x、0.8x、0.85x、0.9x和0.95x的每种除草剂及其所有可推导出的组合,以及如0.97x和0.99x的较高的量。在某些植物健康的实施方式中,可能希望单独使用这些量的麦草畏。或者,由于发现甚至麦草畏的高施用量也不会明显地损伤含有DMO转基因的植物,因此可以使用1x和更高的施用量。可商购的除草剂制剂的制造者设定1x的施用量,这是本领域的技术人员已知的。例如,Fallow MasterTM(含有比例为大约2:1的草甘膦:麦草畏的草甘膦和麦草畏混合物)的标签根据杂草种类和杂草高度,推荐大约451g/ha(311aeg/ha草甘膦:140aeg/ha麦草畏)到621aeg/ha(428aeg/ha草甘膦:193aeg/ha麦草畏)的施用量。草甘膦也可以联合3,6-DCSA施用。通过将植物或植物的部分接触麦草畏,通过利用麦草畏控制杂草,可以获得对于植物健康有益的效应。或者,可以向农作物提供非除草量的麦草畏,但是其仍会产生有益的健康效应。
麦草畏,或3,6-DCSA,或3,6-DCSA的类似物,或通过DMO介导的麦草畏代谢可以产生的产物,可以根据本领域技术人员的知识,例如通过改变施用时间、农作物和/或杂草植物的生长期或施用量,在用于杂草控制的典型施用窗之外施用。确认为需要改善健康的植物可以是经受营养生长和对病害或杂草生长敏感的不成熟的植物,如处于VE到V3或V4的生长阶段的大豆植物。大豆植物也可以处于V4后的生长阶段。植物也可以是处于如VE到V3、V7、V10、V15或VT的生长阶段之前或之中的玉米植物。玉米植物也可以处于营养生长后的生长阶段。植物也可以是经受营养生长的棉花植物,如开花之前,包括,例如出苗、第一真叶、第一营养侧芽、第一果枝或第一棉蕾(果芽)形成的生长阶段之前或期间。棉花植物也可以处于开花的生长阶段或棉桃发育期。也可以以种植后的天数来定义植物的生长阶段。
“麦草畏”指3,6-二氯-邻-茴香酸或3,6-二氯-2-甲氧基苯甲酸及其酸和盐。其盐包括异丙胺盐、二甘醇胺盐、二甲胺盐、钾盐和钠盐。麦草畏的商业制剂的例子包括,但不限于,(作为DMA盐)、(作为DGA盐)、VEL-58-CS-11TM和VanquishTM(BASF,作为DGA盐)。“草甘膦”指N-膦酰基甲基甘氨酸及其盐。草甘膦有多种制剂可商购。草甘膦的这些制剂的例子包括,但不限于,由Monsanto Company销售的 ULTRA、 ULTRAMAX、 CT、 EXTRA、 BIACTIVE、 BIOFORCE、 和除草剂,全部都包含异丙胺盐形式的草甘膦;含有钾盐形式的草甘膦的WEATHERMAX;含有铵盐形式的草甘膦的 DRY和除草剂;含有钠盐形式的草甘膦的GEOFORCE;和含有三甲基锍盐形式的草甘膦的除草剂。
在某些实施方式中,本发明可以涉及DMO转基因的使用。在本发明的一方面,DMO可以由选自下组的核苷酸序列编码:a)编码SEQ ID NO:1的多肽的核酸序列;b)包含SEQ ID NO:2序列的核酸序列;和c)编码与SEQ ID NO:1的多肽具有至少90%的序列同一性的多肽的核酸序列,其中,该多肽具有麦草畏单加氧酶活性,且在对应于SEQ ID NO:1的氨基酸112的位点处包含半胱氨酸。另一方面,核酸序列可以编码与SEQ ID NO:1的多肽具有至少90%的序列同一性的多肽,该多肽具有麦草畏单加氧酶活性,且在对应于SEQID NO:1的氨基酸112的位点处包含色氨酸。在这样的实施方式中,可以提供一种DNA载体,其包含与启动子可操作连接的本文所述的编码DMO的核酸。启动子可以在植物细胞中起作用。在某些实施方式中,编码麦草畏单加氧酶的核酸序列可以与叶绿体转运肽(CTP)可操作地连接。在其它实施方式中,本发明可以涉及使用提供草甘膦耐受性的转基因,如CP4 EPSPS。例如,可以在美国专利RE39,247中发现该基因的序列,本文引入作为参考。
具有降解麦草畏能力的DMO,以及草甘膦耐受基因或其它除草剂耐受基因,能够容易地制备,并根据标准方法分析其活性。例如,也可以通过本领域已知的技术由包括降解除草剂的细菌的合适的生物体鉴定这些序列(美国专利5,445,962;Cork和Krueger,1991;Cork和Khalil,1995)。分离DMO或其它序列的一种方法是,例如,通过与由来源生物体构建的文库的核酸杂交,或通过使用来自来源生物体的mRNA和基于公开序列的引物的RT-PCR。因此,本发明包括使用在严格条件下与本文所述的编码DMO的序列杂交的核酸。本领域的技术人员理解,可以通过增加盐浓度和降低温度来降低条件的严格性。因此,杂交条件容易控制,因而一般是根据想要的结果而选择的方法。高严格条件的一个例子是5X SSC、50%甲酰胺和42℃。通过在这样的条件下进行洗涤,例如,进行10分钟,可以除去那些在这样的条件下不与特定的目标序列杂交的序列。
也可以化学合成变异体,例如,按照本领域内熟知的技术使用已知的DMO多核苷酸进行。例如,可以通过氨基磷酸盐(phosphoroamidite)化学法在自动DNA合成仪中合成DNA序列。化学合成具有许多优点。尤其是,因为可以利用将要表达DNA序列的宿主优选的密码子优化表达,所以化学合成是理想的。并非所有的密码子都需要改变以获得提高的表达,但优选地至少将很少在宿主中使用的密码子改变为宿主优选的密码子。通过改变密码子为宿主优选的密码子可以获得高水平的表达。许多宿主细胞的密码子偏好是已知的(PCT WO 97/31115、PCT WO 97/11086、EP 646643、EP 553494和美国专利第5,689,052、5,567,862、5,567,600、5,552,299和5,017,692号)。其它宿主细胞的密码子偏好可以通过本领域已知的方法推导出。而且,使用化学合成,能够容易地改变DNA分子的序列或其编码的蛋白质以(例如)优化表达(例如,消除干扰转录或翻译的mRNA的二级结构),在方便的点添加独特的限制位点,以及删去蛋白酶酶切位点。
在保持酶活性的同时,可以对本文提供的蛋白质的多肽序列如DMO序列进行修饰和改变。以下是基于改变蛋白质的氨基酸以产生等效的、或甚至是改进的、修饰的多肽和相应的编码序列的讨论。例如,已知某些氨基酸可以替代蛋白质结构中的其它氨基酸,而没有明显损失与底物分子上的结构如结合位点相互结合的能力。因为蛋白质的相互作用能力和性质确定了蛋白质的生物功能活性,所以能够在蛋白质序列中进行某些氨基酸序列取代,当然,也能够对其DNA编码序列进行取代,而仍然能够获得具有类似性质的蛋白质。因此,预期可以在本文所述的DMO肽序列或其它除草剂耐受的多肽和相应的DNA编码序列中进行各种改变,而不明显损失其生物效用或活性。
在进行这些改变时,可以考虑氨基酸的亲水指数(hydropathicindex)。氨基酸亲水指数在使蛋白质具有相互作用生物功能中的重要性一般可为本领域的技术人员所理解(Kyte等人,1982)。可以认为,氨基酸的相对亲水特征对于产生的蛋白质的二级结构有贡献,其二级结构又限定了蛋白质与其它分子例如酶、底物、受体、DNA、抗体、抗原等的相互作用。根据其疏水性和电荷性质,每个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)。
本领域内的技术人员已知,氨基酸可以被具有类似的亲水指数或得分的其它氨基酸取代,并仍然产生具有类似生物活性的蛋白质,即,仍然获得生物功能相当的蛋白质。在进行这样的改变时,优选其亲水指数在±2之内的氨基酸的取代,尤其优选在±1之内的氨基酸的取代,甚至更优选在±0.5之内的氨基酸的取代。
本领域内的技术人员也可以理解,可以根据亲水性有效地进行相似氨基酸的取代。美国专利4,554,101记载,受其邻近的氨基酸的亲水性控制,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,下面的亲水性值已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)。可以理解,一种氨基酸可以取代另一种具有类似的亲水性值的氨基酸,并仍然获得生物功能相当的蛋白质。在这样的变化中,优选其亲水性值在±2之内的氨基酸的取代,尤其优选在±1之内的氨基酸的取代,甚至更优选在±0.5之内的氨基酸的取代。将这些和各种前述性质考虑在内的示例性的取代是本领域的技术人员熟知的,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、以及缬氨酸、亮氨酸和异亮氨酸。
可以在测试系统中,如原生质体、瞬时或稳定转化的植物细胞中,表达包含与DMO序列可操作地连接的CTP序列的DNA构建体,方法是通过将它们可操作地连接于在植物中具有功能性的启动子上。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,PClSV启动子)、SEQ ID NO:41的PClSV启动子、美国专利6,635,806(γ-醇溶蛋白(gamma-coixin)启动子)和美国专利第7,151,204号(玉米叶绿体醛缩酶启动子)。可以使用的另外的启动子为胭脂氨酸合酶(nopaline synthase)(NOS)启动子(Ebert等人,1987)、章鱼氨酸合酶(octopine synthase)(OCS)启动子(其在根癌土壤杆菌(Agrobacterium tumefaciens)的诱导肿瘤的质粒上携带)、花椰菜花叶病毒启动子如花椰菜花叶病毒(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基因复合物启动子(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;美国专利第5,322,938、5,352,605、5,359,142和5,530,196号)、FMV35S(美国专利6,051,753、5,378,619)、花生褪绿条死病花椰菜花叶病毒(peanut chlorotic streak caulimovirus)(PClSV;美国专利5,850,019)、At.Act7(登录号U27811)、At.ANT1(美国专利申请20060236420)、FMV.35S-EF1a(美国专利申请20050022261)、eIF4A10(登录号X79008)和AGRtu.nos(GenBank登录号V00087;Depicker等人,1982;Bevan等人,1983)、水稻胞质磷酸丙糖异构酶(OsTPI;美国专利第7,132,528号)和水稻肌动蛋白15基因(OsAct15;美国专利申请2006-0162010)启动子,可能具有特别的益处。
作为翻译前导序列发挥功能的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,可以包含在启动子和CTP-DMO序列之间。翻译前导序列存在于翻译起始序列的上游的完全加工的mRNA中。翻译前导序列可以影响初级转录物向mRNA的加工、mRNA的稳定性或翻译效率。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(美国专利第5,362,865号)、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(rubisco)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登录号V00087;Bevan等人,1983)等。(Turner和Foster,1995)。在本发明中,尤其发现有益的5’UTR为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(美国专利第7,132,528号)、OsAct15(美国公开第20060162010号)和AGRtunos(GenBank登录号V00087;Bevan等人,1983)。
3’非翻译序列、3’转录终止区或多腺苷酸化区是指连接于结构多核苷酸分子上并位于其下游的DNA分子,包括提供能够影响转录、mRNA加工或基因表达的多腺苷酸化信号和其它调节信号的多核苷酸。多腺苷酸化信号在植物中发挥作用,以引起多腺苷酸核苷酸(polyadenylate nucleotides)向mRNA前体的3’末端的添加。多聚腺苷酸化序列可以源自天然基因、各种植物基因或T-DNA基因。这些序列可以包括在CTP-DMO序列的下游。3’转录终止区的例子为胭脂氨酸合酶3’区(nos 3’;Fraley等人,1983)。不同的3’非翻译区的使用是示例性的(Ingelbrecht等人,1989)。来自豌豆(Pisum sativum)RbcS2基因(Ps.RbcS2-E9;Coruzzi等人,1984)、AGRtu.nos(Genbank登录号E01312)、E6(登录号U30508)和TaHsp17(小麦低分子量热休克蛋白基因;登录号X13431)的多腺苷酸化分子尤其对于本发明的使用有益。
除了上述表达元件,为了表达编码区,在启动子和3’UTR之间可能需要内含子,尤其是在单子叶植物中。“内含子”指可以从基因的基因组拷贝的间插序列中分离或鉴定并且通常定义为在翻译前的mRNA加工过程中被剪接下来的区域的多核苷酸分子。或者,可以通过合成产生内含子。内含子本身可以包含影响可操作连接的基因的转录的亚元件,如顺式元件或增强子结构域。“植物内含子”是在植物细胞中发挥功能的自然或非自然的内含子。植物内含子可用作调节可操作地连接的基因的表达的调整元件。转化构建体中的多核苷酸分子序列可以包含内含子。相对于可转录的多核苷酸分子序列,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5,641,876)、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;美国专利第7,132,528号),且对于实施本发明是有益的。
可以根据本发明使用本领域内任何已知的将转基因构建体引入植物的技术(见,例如,Miki等人,1993)。据信,用于植物转化的合适的方法实际上包括任何能够将DNA引入细胞中的方法,如美国专利第5,384,253号所示的电穿孔,美国专利第5,015,580、5,550,318、5,538,880、6,160,208、6,399,861和6,403,865号所示的微粒轰击(Microprojectile bombardment),如美国专利第5,635,055、5,824,877、5,591,616、5,981,840和6,384,301号所示的土壤杆菌(Agrobacterium)介导的转化,和如美国专利第5,508,184号所示的原生质体转化。通过这些技术的应用,实际上任何植物种类的细胞都可以被稳定地转化,并且这些细胞可以发育为转基因植物。在美国专利第5,846,797、5,159,135、5,004,863和6,624,344中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和美国专利7,002,058中公开了转化大豆的技术。例如,在WO9506722中公开了转化玉米的技术。可用于本发明的植物的一些非限制性的例子包括,苜蓿、大麦、豆类、甜菜、椰菜、卷心菜、胡萝卜、油菜、花椰菜、芹菜、大白菜、玉米、棉花、黄瓜、成熟的菜豆、茄子、茴香、青刀豆、葫芦、韭菜、莴苣、甜瓜、燕麦、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、糖用甜菜、向日葵、柳枝稷、番茄、西瓜和小麦。
在实现外源性的DNA向受体细胞递送之后,产生转基因植物的下一步一般涉及为了进一步的培养和植物再生而鉴别转化的细胞。为了提高鉴别转化体的能力,技术人员可能需要为根据本发明制备的转化载体采用可选择的或可筛选的标记基因。在这种情况下,技术人员随后一般通过将细胞暴露于选择性试剂来分析可能转化的细胞群体,或根据所需的标记基因性状筛选细胞。
暴露于选择性试剂而存活的细胞,或在筛选分析中评分为阳性的细胞,可以在支持植物再生的培养基中培养。可以通过包含如生长调节剂的另外的物质来修饰任何合适的植物组织培养基,例如,MS或N6培养基(Murashige和Skoog,1962;Chu等人,1975)。组织可以保持在含有生长调节剂的基本培养基中,直至可以获得足够的组织来开始植物再生尝试,或进行反复几轮人工选择,直至组织的形态学适于再生,典型地为至少2周,然后转移到有助于幼苗形成的培养基中。定期转移培养物,直至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中。一旦形成足够的根,就将植物转移到土壤中进行进一步的生长和成熟。
为了证实外源性DNA或“转基因”在再生植物中的存在,可以进行多种分析。这样的分析包括,例如,“分子生物学”分析,如DNA印迹法和RNA印迹法与PCRTM;“生物化学”分析,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),或通过酶功能;植物部分分析,如叶或根分析;以及,通过分析整个再生植物的表型。
一旦转基因已引入植物中,就能够通过杂交将基因引入任何与第一种植物性相容的植物中,而无需直接转化第二种植物。因此,本文所用的术语“后代”是指根据本发明制备的亲本植物的任一代的后代,其中,后代包含根据本发明制备的选择的DNA构建体。“转基因植物”因而可以是任一代。如本文所公开的,“杂交”植物以提供相对于起始植物系具有一个或多个添加的转基因或等位基因的植物系,被定义为通过将起始系与包含本发明的转基因或等位基因的供体植物系杂交从而将特定序列引入植物系的技术。为了实现这一点,例如,技术人员可以进行以下步骤:(a)种植第一(起始系)和第二(包含所需转基因或等位基因的供体植物系)亲本植物的种子;(b)使第一和第二亲本植物的种子生长为有花的植物;(c)用第二亲本植物的花粉对第一亲本植物的花授粉;和(d)收获具有已授粉的花的亲本植物上产生的种子。
提供对一种或多种除草剂的耐受性的未修饰或修饰的蛋白质分子及其相应的核酸分子在本领域内是熟知的。例如:
a)编码草甘膦耐受性的序列包括5-烯醇丙酮酰莽草酸(enolpyruvylshikimate)-3-磷酸酯合酶(EPSPS;美国专利5,627,061、美国专利RE39,247、美国专利6,040,497、美国专利5,094,945、WO04074443和WO04009761)、草甘膦氧化还原酶(GOX;美国专利5,463,175)、草甘膦脱羧酶(WO05003362和美国专利申请20040177399)和草甘膦-N-乙酰基转移酶(GAT;美国专利申请20030083480),给予对草甘膦的耐受性;
b)麦草畏单氧合酶(DMO,由ddmC编码),给予对植物生长素样除草剂如麦草畏的耐受性(美国专利申请公开20030115626、20030135879;Wang等人,1996;Herman等人,2005);
c)膦丝菌素乙酰转移酶(phosphinothricin acetyltransferase)(bar),给予对膦丝菌素或草丁膦的耐受性(U.S.5,646,024、U.S.5,561,236、EP 275,957、U.S.5,276,268、U.S.5,637,489、U.S.5,273,894);
d)2,2-二氯丙酸脱卤素酶,给予对2,2-二氯丙酸(茅草枯(Dalapon))的耐受性(WO9927116);
e)乙酰羟酸合酶或乙酸乳酸合酶,给予对如磺酰脲、咪唑啉酮、三唑嘧啶、嘧啶氧苯甲酸酯和2-苯并[c]呋喃酮的乙酰乳酸合酶抑制剂的耐受性(U.S.6,225,105、U.S.5,767,366、U.S.4,761,373、U.S.5,633,437、U.S.6,613,963、U.S.5,013,659、U.S.5,141,870、U.S.5,378,824、U.S.5,605,011);
f)卤素芳基腈水解酶(Bxn),给予对溴苯腈的耐受(WO8704181A1、U.S.4,810,648、WO8900193A);
g)修饰的乙酰辅酶A羧化酶,给予对环己烯二酮(稀禾定(sethoxydim))和芳氧基苯氧基丙酸酯(吡氟氯禾灵(haloxyfop))的耐受性(U.S.6,414,222);
h)二氢蝶酸合酶(dihydropteroate synthase)(sulI),给予对磺胺除草剂的耐受性(U.S.5,597,717、U.S.5,633,444、U.S.5,719,046);
i)32kD光系统II多肽(psbA),给予对三嗪除草剂的耐受性(Hirschberg等人,1983);
j)邻氨基苯甲酸合酶,给予对5-甲基色氨酸的耐受性(U.S.4,581,847);
k)二氢吡啶二羧酸合酶(dapA),给予对氨乙基半胱氨酸的耐受性(WO8911789);
l)八氢番茄红素去饱和酶(crtI),给予对如氟草敏(norflurazon)的哒嗪酮除草剂的耐受性(JP06343473);
m)羟基-苯基丙酮酸双加氧酶,给予对如异噁唑草酮的环丙基异恶唑类(cyclopropylisoxazole)除草剂的耐受性(WO 9638567;U.S.6,268,549);
n)修饰的原卟啉原氧化酶I(protox),给予对原卟啉原氧化酶抑制剂的耐受性(U.S.5,939,602);和
o)芳氧基链烷酸双加氧酶(AAD-1),给予对含有芳氧基链烷酸部分的除草剂的耐受性(WO05107437)。这样的除草剂的例子包括苯氧基植物生长素(如2,4-D和2,4-滴丙酸(dichlorprop))、吡啶氧基植物生长素(如氟草烟(fluroxypyr)和绿草定(triclopyr))、芳氧基苯氧基丙酸(AOPP)乙酰辅酶A羧化酶(ACCase)抑制剂(如吡氟氯禾灵(haloxyfop)、喹禾灵(quizalofop)和禾草灵(diclofop))和5-取代的苯氧基乙酸原卟啉原氧化酶IX抑制剂(如吡草醚(pyraflufen)和氟烯草酸(flumiclorac))。
实施例
包括下面的实施例,以说明本发明的实施方式。本领域的技术人员应该理解,以下实施例中公开的技术代表发明人发现的在本发明的实施中作用良好的技术。但是,根据本公开内容,本领域的技术人员应该理解,在不背离本发明的概念、精神和范围的情况下,对公开的具体实施例可以进行许多改变,并仍然能够获得类似的结果。更具体地说,在化学和生理学上都相关的某些试剂显然可以取代本文描述的试剂,而能够获得相同或类似的结果。所有这些对于本领域技术人员来说显而易见的类似的取代和修饰,都被视为在由附加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
含有DMO基因的转基因大豆的产生
使用标准程序(例如,U.S.6,384,301、U.S.7,002,058或Zhang等人,1999),通过用包含编码SEQ ID NO:1的多肽的编码DMO的多核苷酸的二元载体对来自栽培品种Thorne、NE3001和A3525的大豆组织进行土壤杆菌介导的转化,获得包含编码DMO的转基因的转基因大豆植物。
实施例2
麦草畏和DCSA对与抗病力有关的病原体相关蛋白质的诱导
将转基因和对照大豆种子(栽培品种Thorne、NE3001和A3525)种植到含有Redi-earthTM(Scotts-Sierra Horticultural Products Co.,Marysville,Ohio)的3.5英寸的方形塑料罐中。在试验持续期间,将罐放置于用于从上部和/或下部灌溉的35英寸x60英寸的玻璃纤维加水盘中的毛细管衬垫上,以维持对于植物生长最佳的土壤湿度,并用(14-14-14缓慢释放;Scotts-Sierra HorticulturalProducts Co.,Marysville,Ohio)以100gm/cu.ft.的量施肥,以在温室试验期间保持植物生长。
植物在温室中生长,条件为29°/21℃的白天/夜晚温度,15%-50%的相对湿度,以模拟晚春的温暖季节的生长条件。如需要,为14小时的最小光周期提供大约600μE(微Einsteins)的补充光照。以根据施用量随机化的随机分组设计建立试验,每个处理重复4-6次。
使用气压设定为最小24psi(磅/平方英寸)或165kpa(千帕斯卡)的 9501E扇形雾锥喷嘴(Spraying Systems Co,Wheaton,IL),用轨道喷雾器(track sprayer)进行所有的除草剂施用。保持喷嘴的高度在进行喷施的植物材料的顶部以上大约16英寸。喷施体积是10加仑/英亩或93升/公顷。当植物到达V-3阶段时,进行施用。
培育在其基因组中携带DMO转基因的植株(event),并在V3阶段后用1lb/英亩的麦草畏(BASF)处理。在处理之后0、3、8、24、48、72小时(HAT)收获叶样品,并冷冻用于进一步分析。在另一实验中,以0.25、0.5和1lb/英亩的量,对含有DMO的植物喷施麦草畏或DCSA,并在处理之后24小时收集组织样品。提取RNA并使用PR-2(即,β-1,3-葡聚糖酶)作为探针通过RNA印迹分析进行分析。PR-2是与发病机理相关的蛋白质,已知其降解真菌膜并因此提供对抗真菌病原体的保护(U.S.5,670 706)。
在含有液氮和玻璃/金属珠的FalconTM管(BD Biosciences,Franklin Lakes,NJ)中磨碎组织样品。将子样品放在埃彭道夫微量离心管(Eppendorf tube)中。按照制造者的规程(Qiagen,Valencia,CA),使用植物RNA小型试剂盒来提取RNA。使用在260和280nm的OD测量值估计RNA浓度。通过加入1/10体积的3M醋酸钠和2.5倍体积的100%乙醇,并将微量离心管在-20℃存储48小时,沉淀5μg(微克)RNA。将微量离心管在4℃离心20分钟,并弃去上清液。向微量离心管中加入70%的乙醇,轻轻地使RNA再悬浮于其中。然后离心微量离心管3.5分钟,干燥RNA沉淀物大约30-60分钟,并通过短暂的涡旋和混合而再悬浮于15μl的加样缓冲液(来自Ambion,Austin,TX的凝胶加样缓冲液II)中。
在65℃将RNA样品和RNA标准(RNA梯;Invitrogen,Carlsbad,CA)变性10分钟,并保持在冰上直至加样于在1x MOPS缓冲液中由1%琼脂糖和2%甲醛制成的凝胶中。在1x MOPS缓冲液中、在17V下使凝胶电泳16小时,并用溴化乙锭染色以显示凝胶中的RNA样品和标准。在水中洗涤凝胶30分钟,以除去溴化乙锭,接着在20xSSC(NaCl和柠檬酸钠)中洗涤两次,每次15分钟。
使用快速向下转移系统(TurblotterTM和印迹组件,Whatman-Schleicher & Schuell;Florham Park,NJ),将RNA样品和标准在膜 PLUS,Midwest Scientific,Valley Park,MO)上印迹5-6小时。用水冲洗膜,紫外交联并置于纸上干燥。在50℃下,膜在温暖的15ml DIG Easy Hyb缓冲液(Roche,Penzberg,德国)中预杂交30分钟。根据使用者的说明书(Roche),用DIG标记PR-2探针,使用PCR DIG探针合成试剂盒合成PR-2探针,煮沸5分钟,并在加到温暖的15μl DIG Easy Hyb缓冲液中之前在冰上冷却。在50℃下进行杂交过夜。在高严格条件下洗涤RNA印迹两次,分别为在室温下,在2x SSC和0.1% SDS溶液中进行5和15分钟。在68℃下,在0.5x SSC和0.1% SDS溶液中进行另外两次洗涤,每次30分钟。然后在1x洗涤缓冲液(Roche)中洗涤印迹2分钟,并在室温下伴随适当的摇动使用封闭试剂(Roche)封闭30分钟到3小时。将印迹转移到30ml DIG抗体溶液(75mU/ml,Roche)中30分钟。通过使用1x洗涤缓冲液洗涤3次,每次15分钟,除去未结合的抗体。在1x检测缓冲液(Roche)中平衡印迹3分钟。将准备使用的CDP-STAR(2-氯-5-(4-甲氧基螺{1,2-二氧杂环丁烷-3,2′-(5′-氯)三环[3.3.1.13,7]癸烷}-4-基)-1-苯基磷酸二钠;Roche)添加到放于检测袋中的印迹上。将袋密封,并将印迹暴露于BIOMAX胶片(Kodak)1.5小时。
图1显示在用麦草畏处理后的各个时间点的PR2的诱导。峰值诱导在48小时处。在没有用麦草畏处理的植物中没有PR2的诱导。
图2显示在用各种量的麦草畏和DSCA处理两种独立的大豆转基因系24小时之后的PR2的诱导。在没有用麦草畏或DSCA处理或只用水处理的植物中没有PR2的诱导。
实施例3
DCSA提供真菌病抗性
将Roundup Weather (Monsanto,St.Louis,MO)和3,6-DCSA(BASF)的混合物喷施于生长10天的Roundup Flex棉花(植株MON 88913;美国公开20060059590)幼苗。对照植物只喷施水。在喷施24小时后,通过将幼苗浸入接种物浆中,用终极腐霉接种两种处理的棉花幼苗。阳性对照植物既没有喷施,也没有接种终极腐霉。如图3所示,接种终极腐霉但是没有接受RoundupWeather (Monsanto,St.Louis,MO)和3,6-DCSA(BASF)混合物施用的棉花幼苗在接种后10天之内由于感染终极腐霉而死亡。但是,在接受Roundup Weather (Monsanto,St.Louis,MO)和3,6-DCSA(BASF)的混合物的植物中没有发生终极腐霉感染。用和DCSA处理的棉花植物与未接种终极腐霉的植物一样健康。结果表明,当用DCSA处理24小时之后用腐霉接种植物时,Roundup Ultra Max和DCSA的联合处理减少了终极腐霉感染。
实施例4
DCSA提供细菌病抗性
在这一实施例中,给生长2周的棉花植物喷施DSCA,而对照植物只接受水的喷施。在DCSA喷施大约24小时后,使用牙签接种法(tooth pick inoculation method)给植物接种细菌性斑点病病原体(野油菜黄单胞菌锦葵致病变种)。如图4所示,接种细菌病原体但没有喷施DSCA的棉花植物发展为坏死斑,这是细菌性斑点病的典型症状。但是,在接种细菌病原体24小时之前接受DCSA施用的棉花植物只显示局部病变(HR应答),且细菌性斑点病的症状没有增加。该实验的结果表明,当在DCSA喷施24小时之后接种细菌病原体时,DCSA施用减少了棉花植物的细菌感染。
实施例5
麦草畏或DCSA增加了大豆对氧化应激的耐受性
对于该实施例,植物如上述实施例所述生长。在其基因组中携带DMO基因的转基因和非转基因大豆植物或者用百草枯(Paraquat)Syngenta)处理以产生氧化应激,或者用麦草畏处理接着在V3阶段用百草枯处理。麦草畏的施用量为1lb/英亩或1120gae/ha。以30、70或200g ae/ha施用百草枯。然后在处理之后的特定的日子(DAT),通过目视评估损伤来评估植物的百草枯损伤,评估等级是相对于未处理的对照植物的0-100%,零代表“没有”损伤,100%代表“完全”损伤或死亡。收集数据并使用合适的统计方法分析。
表1显示在对携带DMO基因并且在施用百草枯24小时之前用麦草畏处理的大豆植物测试的所有三个施用量上,在处理后4天可见明显减少的百草枯损伤。
表1.用百草枯或麦草畏处理,接着在V-3阶段用百草枯处理的非转基因或转基因大豆植物的损伤百分比。以ANOVA平均值比较表示损伤百分比(相对于百草枯处理)。类似的字母代表在p=0.05水平上没有统计学差异。
在另一实验中,测试外部施用的DSCA在保护大豆植物免受百草枯导致的氧化应激方面的益处。在施用百草枯24小时之前施用不同量的DSCA,以测试施用的DSCA的量和如上所述根据损伤百分比减少确定的氧化应激保护程度之间的任何相关性。也分开施用不同水平的麦草畏,以测试相同的假设。
表2显示,例如,麦草畏或DSCA的施用提供了针对由随后施用百草枯导致的氧化应激的保护。接触单独的百草枯的转基因植物显示大约75%的损伤比,而接触不同量的麦草畏或DSCA接着接触百草枯的转基因植物令人惊奇地显示损伤减少。此外,损伤的减少程度随着麦草畏或DSCA的施用量的增加而增加。该实验使用的植物是转基因的。麦草畏的效应是由于麦草畏在植物中转化为DCSA或随后的代谢,而DCSA的效应是由于DCSA本身或其代谢物。
表2.仅用百草枯处理,或用麦草畏或DSCA处理接着在V-3阶段用百草枯处理的转基因大豆植物的损伤百分比。以ANOVA平均值比较表示损伤百分比。
实施例6
DCSA增加了棉花对氧化应激的耐受性
如表3所示的4种不同的棉花栽培品种在4-英寸的罐中生长2周。这些棉花植物或者喷施1lb/A的量的DSCA(BASF),或者不喷施(没有DSCA施用,作为对照)。在施用DSCA24小时后,将百草枯(Syngenta)以30gm/ha、70gm/ha和100gm/ha的量喷施于实验植物(用DSCA处理的或未处理的),以模拟氧化应激。阳性对照植物没有接受任何化学处理(既没有施用DCSA也没有施用Gramoxone)。然后在用处理2天之后(DAT),通过目视评估来评价所有植物的百草枯损伤,评价的等级是相对于未处理的对照植物的0-100%,零代表“没有”损伤,100%代表植物“完全”损伤或死亡。在棉花的两个不同生长阶段(2节阶段和5节阶段)重复相同的实验。
表3所示的这些实验的结果表明,当RR棉花的4个品种在两个不同的生长阶段(2节阶段和5节阶段)用DSCA预处理(24小时)时,Gramoxone损伤明显减少。损伤症状的减少不依赖于测试的棉花的两个生长阶段。非转基因棉花栽培品种ST474获得类似的结果。
表3.用百草枯或DCSA处理,接着用百草枯处理的所示棉花植物品种的损伤百分比
ST4646B2RF | ST3273B2RF | ST4575BR | ST6622RF | |
未处理的对照 | 0 | 0 | 0 | 0 |
Gramoxone(30ga.i./ha) | 31.5 | 35 | 37.5 | 42.5 |
DCSA(1lb/英亩)+Gramoxone(30ga.i./ha) | 8.5 | 6 | 12.5 | 7.5 |
Gramoxone(70ga.i./ha) | 42.5 | 50 | 42.5 | 50 |
DCSA(1lb/英亩)+Gramoxone(70ga.i./ha) | 20 | 32.5 | 30 | 35 |
Gramoxone(100ga.i./ha) | 70 | 50 | 65 | 70 |
DCSA(1lb/英亩)+Gramoxone(100ga.i./ha) | 45 | 45 | 40 | 50 |
实施例7
通过向植物施用麦草畏提高产量
通过培育非转基因和转基因植物,向转基因植物施用麦草畏并收获种子,测试DSCA在提高产量方面的益处。在生长季节开始阶段前后,在如土壤湿度、温度和播种深度等可能的最佳生长条件下,在几个位置种植非转基因和转基因大豆种子。在所有位置,按分区(split-plot)设计种植种子,麦草畏处理作为全区(whole-plot)效应,植株作为分区效应。设计细节如下:6个位置、2个重复/位置、2排/区,排的长度为12英尺(+3英尺狭径(alley))、9个种子/英尺、108个种子/排、5个植株(植株1-4和分开的第五植株);和如表4所示的4个处理。总共在6个位置种植240个区(每个位置40个)。
表4.应用于转基因大豆的4个麦草畏处理的细节
用一种已知的商业系,如A3525,在实验区的周围种植4个非转基因边行。遵循本领域内已知的最佳生产和管理实践。按需要进行最大害虫控制和病害控制,以防止麦草畏施用的混合效应。根据标准方法按需灌溉田地。
如图5所示的结果表明,DMO转基因的存在没有给未用麦草畏处理的大豆带来产量损失或其它农业学上的效应。相反地,如图6所示,在4/5的植株和13/15的处理中,相对于未喷施的对照,麦草畏喷施显示增加或相当的产量,表明由于通过DMO对麦草畏的作用体内产生的DCSA改善了植物健康,导致产量的益处。
实施例8
通过表达麦草畏和草甘膦耐受性基因并用草甘膦和麦草畏处理植物,提供多重益处
已发现如CP4 EPSPS的草甘膦耐受性基因提供草甘膦耐受性,并通过提供针对包括豆薯层锈菌(Phakopsora pachyrizi)和山马蝗层锈菌(Phakopsora meibomiae)(大豆锈病的病原体)的几种病原体的抗性改善植物健康,这种抗性是通过草甘膦对这些病原真菌的直接作用而提供的(WO05102057)。在目前情况下,显示DMO提供了麦草畏耐受性,并通过减少病原体和氧化应激来改善植物健康。通过分子和育种方法将两种基因结合到一个植物中,并用草甘膦和麦草畏处理植物,能够获得进一步的益处。这会增加取决于其市场和环境需要的种植者可采用的选择的数目。
实施例9
DCSA对于棉花根癌线虫(RKN;根结线虫)的控制
出乎意料地发现DSCA控制虫瘿的形成、增加植物高度并减少棉花植物(ST474)中线虫产卵的数量(表5)。在用RKN卵对生长10-14天的棉花植物接种之前24小时,用DSCA以1lb/a的量处理它们。与没有用DSCA处理的植物相比,用DSCA处理的棉花植物显示没有或较低的虫瘿形成(0.6比2.6)、增加的植物高度(19.8cm比16.9cm)和减少的卵计数(4575比6425)。在接种45天后,测量植物高度、虫瘿等级和卵计数。虫瘿等级尺度为0-5,其中,0=没有可见的虫瘿形成,5=严重的虫瘿形成。
表5.在棉花中施用DSCA对RKN的控制
实施例10
DCSA提供针对轮枝孢属的保护
出乎意料地发现DSCA为棉花植物提供了针对棉花枯萎病的病原体大丽花轮枝孢(Verticillium dahliae)的保护(表6)。通过幼苗浸入方法,用轮枝孢属的温和分离株(DPL)或毒力分离株(King)对品种ST6611、BOLLGARD 2、ROUNDUP READY FLEX的棉花植物进行接种。从罐中取出处于第一真叶期的幼苗,切下一英寸的根,将幼苗浸入孢子浆中一分钟,然后转移回罐中。这些菌株由TerryWheeler博士(Texas A&M University,TX USA)分离并提供。在接种24小时之前,以1lb/A的量施用DSCA和以0.75lb/A的量施用草甘膦(ROUNDUP WEATHERMAX,Monsanto)。按1-9的病害严重性等级,检测针对轮枝孢菌属的保护:1为健康,9为由于病害而死亡,也以10个接种的植物中存活的植物数来检测。DSCA的施用提供了针对大丽花轮枝孢的中度和强度分离株的保护。相比没有用DSCA处理的接种的检查株,用DSCA处理的接种的棉花植物显示较低的等级和较高的存活等级。
表6.对棉花施用DCSA对轮枝孢菌属的控制
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根据本公开内容,不需要过多的实验就可以制备和实施本文公开的和要求保护的所有组合物和/或方法。尽管已结合优选的实施方式描述了本发明的组合物和方法,但在不背离本发明的概念、精神和范围的情况下,可以对本文所述的组合物和/或方法和方法的步骤或步骤顺序进行变化,这对于本领域的技术人员是显而易见的。更具体地说,化学和生理学上都相关的某些试剂显然可以取代本文描述的试剂,而可以实现相同或类似的结果。所有这些对于本领域技术人员是显而易见的类似的取代和改变都被视为在由附加的权利要求所限定的本发明的精神、范围和概念之内。
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序列表
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FENG,PAUL
PIIKIN,JOHN
HOI,SIO-WAI
<120>改善植物健康的方法和组合物
<130>MONS:116WO
<140>未知
<141>2007-010-15
<150>60/852,308
<151>2006-010-16
<160>2
<170>PatentIn Ver.2.1
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<211>340
<212>PRT
<213>人工序列
<220>
<223>人工序列的描述:合成的肽
<400>1
<210>2
<211>1023
<212>DNA
<213>人工序列
<220>
<223>人工序列的描述:合成的引物
<400>2
Claims (50)
1.一种改善植物健康的方法,包括向植物提供一定量的DMO所介导代谢的产物或其类似物,与没有向其提供麦草畏或DMO所介导代谢的产物或其类似物的相同基因型的植物相比,所述量改善植物的健康。
2.如权利要求1所述的方法,包括向植物提供一定量的3,6-DCSA或其类似物,与没有向其提供3,6-DCSA或其类似物的相同基因型的植物相比,所述量改善植物的健康。
3.如权利要求1所述的方法,其中,所述植物在农作物生产田地中。
4.如权利要求1所述的方法,进一步包括在向植物提供DMO所介导代谢的产物或其DMO介导的代谢产物或其类似物之前、同时或之后,植物遭受生物性应激或非生物性应激。
5.如权利要求1所述的方法,进一步包括在向植物提供麦草畏或DMO所介导代谢的产物或其类似物之前,确认植物需要改善植物健康的步骤。
6.如权利要求5所述的方法,其中,确认植物需要改善植物健康包括确认植物显示至少指示生物性应激或非生物性应激的第一症状。
7.如权利要求6所述的方法,其中,所述症状选自矮化、光合作用功能丧失、脂质过氧化作用、活性氧物质的积聚、自由基含量增加和坏死。
8.如权利要求1所述的方法,其中,所述植物处于非生物性应激的危险或在非生物性应激之下。
9.如权利要求8所述的方法,其中,所述非生物性应激选自渗透应激、热或寒冷暴露、氧化应激和营养缺乏。
10.如权利要求1所述的方法,其中,所述植物处于生物性应激的危险或在生物性应激之下。
11.如权利要求10所述的方法,其中,所述生物性应激选自真菌病、病毒病、细菌病、昆虫侵扰、线虫侵扰和杂草侵扰。
12.如权利要求11所述的方法,其中,所述生物性应激是大豆锈病、黄萎病、细菌性黑枯病、幼苗病或根癌线虫侵扰。
13.如权利要求1所述的方法,其中,所述植物对氧化应激的耐受性增强。
14.如权利要求1所述的方法,进一步限定为包括向植物群提供所述麦草畏或其DMO介导的代谢产物或其类似物,以改善植物的健康。
15.如权利要求1所述的方法,其中,所述DMO所介导代谢的产物是3,6-DCSA、3,5-DCSA、DCGA或3-CSA,或3,6-DCSA、3,5-DCSA、DCGA或3-CSA的代谢物。
16.如权利要求1所述的方法,其中,所述DMO所介导代谢的产物是3,6-DCSA的类似物。
17.如权利要求1所述的方法,其中,所述植物包含编码DMO的转基因。
18.如权利要求1所述的方法,其中,所述产物不是除草性的。
19.如权利要求4所述的方法,包括用大约0.25lb/a到大约1.5lb/a的麦草畏处理所述田地。
20.如权利要求4所述的方法,包括用大约0.25lb/a到大约12lb/a的DCSA处理所述田地。
21.如权利要求1所述的方法,其中,所述植物不包含编码DMO的转基因。
22.如权利要求1所述的方法,其中,所述植物是双子叶植物。
23.如权利要求22所述的方法,其中,所述双子叶植物选自苜蓿、豆类、甜菜、椰菜、卷心菜、胡萝卜、花椰菜、芹菜、大白菜、棉花、黄瓜、茄子、亚麻、莴苣、羽扇豆、甜瓜、豌豆、胡椒、花生、马铃薯、南瓜、萝卜、油菜籽、菠菜、大豆、倭瓜、糖用甜菜、向日葵、番茄和西瓜。
24.如权利要求1所述的方法,其中,所述植物是单子叶植物。
25.如权利要求24所述的方法,其中,所述单子叶植物选自大麦、玉米、韭菜、洋葱、水稻、高梁、甜玉米、小麦、黑麦、小米、甘蔗、燕麦、黑小麦、柳枝稷和草坪草。
26.如权利要求1所述的方法,其中,进一步限定所述植物耐受选自草甘膦、草丁膦、2,4-D、硝草酮、噻草定、异噁唑草酮、溴苯腈、莠去净、精吡氟禾草灵和磺酰脲类/咪唑啉酮类的除草剂。
27.如权利要求26所述的方法,进一步包括将植物接触选自所述草甘膦、草丁膦、2,4-D、硝草酮、噻草定、异噁唑草酮、溴苯腈、莠去净、精吡氟禾草灵和磺酰脲类/咪唑啉酮类的至少一种其它除草剂。
28.如权利要求27所述的方法,其中,接触至少一种其它除草剂发生在接触麦草畏或其DMO介导的代谢产物之前。
29.如权利要求27所述的方法,其中,接触至少一种其它除草剂与接触麦草畏或其DMO介导的代谢产物同时发生。
30.如权利要求27所述的方法,其中,接触至少一种其它除草剂发生在接触麦草畏或其DMO介导的代谢产物之后。
31.如权利要求27所述的方法,包括将植物接触草甘膦。
32.如权利要求2所述的方法,其中,所述植物在营养生长期。
33.如权利要求32所述的方法,其中,所述植物是处于VE-V4生长阶段的大豆植物。
34.一种提高植物产量的方法,包括将植物与一定量的麦草畏或包括DCSA在内的DMO介导的麦草畏代谢的产物接触,相对于在同样的条件下生长但不接触麦草畏或DMO介导的麦草畏代谢产物的相同基因型的植物,所述量有效提高植物的产量。
35.如权利要求34所述的方法,其中,所述植物在农作物生产田地中。
36.如权利要求34所述的方法,进一步限定为包括将植物群接触所述麦草畏或其DMO介导的代谢产物。
37.如权利要求34所述的方法,其中,所述DMO介导的代谢产物是3,6-DCSA、3,5-DCSA或3-CSA,或3,6-DCSA、3,5-DCSA或3-CSA的代谢物。
38.如权利要求34所述的方法,其中,所述DMO介导的代谢产物是DCSA的类似物。
39.如权利要求34所述的方法,其中,所述产物是除草性的;并且,所述植物包含编码DMO的转基因。
40.如权利要求34所述的方法,其中,所述产物不是除草性的。
41.如权利要求35所述的方法,包括用大约0.25lb/a到大约1.5lb/a的麦草畏处理所述田地。
42.如权利要求35所述的方法,包括用大约0.25lb/a到大约12lb/a的DCSA处理所述田地。
43.如权利要求34所述的方法,其中,所述植物是双子叶植物。
44.如权利要求43所述的方法,其中,所述双子叶植物选自苜蓿、豆类、甜菜、椰菜、卷心菜、胡萝卜、花椰菜、芹菜、大白菜、棉花、黄瓜、茄子、亚麻、莴苣、羽扇豆、甜瓜、豌豆、胡椒、花生、马铃薯、南瓜、萝卜、油菜籽、菠菜、大豆、倭瓜、糖用甜菜、向日葵、番茄和西瓜。
45.如权利要求34所述的方法,其中,所述植物是单子叶植物。
46.如权利要求45所述的方法,其中,所述单子叶植物选自大麦、玉米、韭菜、洋葱、水稻、高粱、甜玉米、小麦、黑麦、小米、甘蔗、燕麦、黑小麦、柳枝稷和草坪草。
47.如权利要求34所述的方法,其中,限定所述植物耐受选自草甘膦、草丁膦、2,4-D、硝草酮、氟硫草定、异噁唑草酮、溴苯腈、莠去净、精吡氟禾草灵和磺酰脲类/咪唑啉酮类的除草剂。
48.如权利要求34所述的方法,进一步包括将植物接触选自草甘膦、草丁膦、2,4-D、硝草酮、噻草定、异噁唑草酮、溴苯腈、莠去净、精吡氟禾草灵和磺酰脲类/咪唑啉酮类的至少一种除草剂。
49.一种生产3,6-DCSA的方法,包括将农作物生产田地中的植物群体与麦草畏接触,其中,所述植物包含转基因编码的DMO活性。
50.一种改善种子的健康的方法,包括将种子与一定量的麦草畏或其DMO介导的代谢产物接触,与不接触麦草畏或其DMO介导的代谢产物的相同基因型的种子相比,所述量改善种子的发芽。
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US10104848B2 (en) | 2015-06-25 | 2018-10-23 | Monsanto Technology Llc | Soybean variety 01051773 |
US9807962B2 (en) | 2015-06-25 | 2017-11-07 | Monsanto Technology Llc | Soybean variety 01058613 |
US9781897B2 (en) | 2015-06-25 | 2017-10-10 | Monsanto Technology Llc | Soybean variety 01058741 |
US9986704B2 (en) | 2015-06-25 | 2018-06-05 | Monsanto Technology Llc | Soybean variety 01057162 |
US10051807B2 (en) | 2015-06-28 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01058772 |
US9907265B2 (en) | 2015-06-28 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058380 |
US9844200B2 (en) | 2015-06-28 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01058749 |
US9974269B2 (en) | 2015-06-28 | 2018-05-22 | Monsanto Technology Llc | Soybean variety 01058766 |
US9826703B2 (en) | 2015-06-28 | 2017-11-28 | Monsanto Technology Llc | Soybean variety 01058683 |
US9820458B2 (en) | 2015-06-28 | 2017-11-21 | Monsanto Technology Llc | Soybean variety 01052110 |
US9848569B2 (en) | 2015-06-28 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01057209 |
US9807963B2 (en) | 2015-06-28 | 2017-11-07 | Monsanto Technology Llc | Soybean variety 01052094 |
US9844199B2 (en) | 2015-06-28 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01058475 |
US9924677B2 (en) | 2015-06-28 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01052136 |
US9907266B2 (en) | 2015-06-28 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057255 |
US9924676B2 (en) | 2015-06-28 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01057057 |
US9924679B2 (en) | 2015-06-29 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01057010 |
US9924678B2 (en) | 2015-06-29 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01057325 |
US9907268B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057333 |
US9907269B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01050536 |
US9901057B2 (en) | 2015-06-29 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01050566 |
US9968057B2 (en) | 2015-06-29 | 2018-05-15 | Monsanto Technology Llc | Soybean variety 01058336 |
US9907270B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057497 |
US9907267B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057383 |
US9844201B2 (en) | 2015-06-29 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01050745 |
US9907271B2 (en) | 2015-06-29 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01056960 |
US10034445B2 (en) | 2015-06-29 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01050611 |
US9901056B2 (en) | 2015-06-29 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01057377 |
US10034444B2 (en) | 2015-06-30 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01050609 |
US9820459B2 (en) | 2015-06-30 | 2017-11-21 | Monsanto Technology Llc | Soybean variety 01058808 |
US9814207B2 (en) | 2015-06-30 | 2017-11-14 | Monsanto Technology Llc | Soybean variety 01058565 |
US9907263B2 (en) | 2015-06-30 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01050612 |
US10004197B2 (en) | 2015-06-30 | 2018-06-26 | Monsanto Technology Llc | Soybean variety 01050595 |
US9826702B2 (en) | 2015-06-30 | 2017-11-28 | Monsanto Technology Llc | Soybean variety 01051835 |
US10070606B2 (en) | 2015-06-30 | 2018-09-11 | Monsanto Technology Llc | Soybean variety 01058789 |
US9907264B2 (en) | 2015-06-30 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01052085 |
US9901055B2 (en) | 2015-06-30 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01050596 |
US9832965B2 (en) | 2015-06-30 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01051604 |
US9901059B2 (en) | 2015-09-24 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01051956 |
US9894862B2 (en) | 2015-09-24 | 2018-02-20 | Monsanto Technology Llc | Soybean variety 01058666 |
US9907273B2 (en) | 2015-09-24 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01051571 |
US9901060B2 (en) | 2015-09-24 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01051946 |
US9907274B2 (en) | 2015-09-24 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01052030 |
US9901061B2 (en) | 2015-09-24 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01052008 |
US9901062B2 (en) | 2015-09-24 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01052029 |
US10563220B2 (en) | 2015-12-21 | 2020-02-18 | Monsanto Technology Llc | Compositions and methods for efficient targeting of transgenes |
US9700000B1 (en) | 2016-03-21 | 2017-07-11 | Monsanto Technology Llc | Soybean variety XR38W15X |
US9596818B1 (en) | 2016-03-21 | 2017-03-21 | Monsanto Technology Llc | Soybean variety XR30AX15X |
US9763413B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR23AQ15X |
US9861055B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR39U15X |
US9565822B1 (en) | 2016-03-21 | 2017-02-14 | Monsanto Technology Llc | Soybean variety XR37AE15X |
US9585355B1 (en) | 2016-03-21 | 2017-03-07 | Monsanto Technology Llc | Soybean variety XR40AA15X |
US9763414B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR25AL15X |
US9763412B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR09AP15X |
US9861054B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR36Z15X |
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 |
US9560828B1 (en) | 2016-03-21 | 2017-02-07 | Monsanto Technology Llc | Soybean variety XR46Z15X |
US9699999B1 (en) | 2016-03-21 | 2017-07-11 | Monsanto Technology Llc | Soybean variety XR22K15X |
US9585354B1 (en) | 2016-03-21 | 2017-03-07 | Monsanto Technology Llc | Soybean variety XR48C15X |
US9788508B1 (en) | 2016-03-21 | 2017-10-17 | Monsanto Technology Llc | Soybean variety XR20AH15X |
US10010040B1 (en) | 2016-03-21 | 2018-07-03 | Monsanto Technology Llc | Soybean variety XR50E15X |
US9763411B1 (en) | 2016-03-21 | 2017-09-19 | Monsanto Technology Llc | Soybean variety XR09AL15X |
US9730415B1 (en) | 2016-03-21 | 2017-08-15 | Monsanto Technology Llc | Soybean variety XR08A15X |
US9861056B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR56J15X |
US9861053B1 (en) | 2016-03-21 | 2018-01-09 | Monsanto Technology Llc | Soybean variety XR36AC15X |
US9560829B1 (en) | 2016-03-21 | 2017-02-07 | Monsanto Technology Llc | Soybean variety XR47AG15X |
US9565821B1 (en) | 2016-03-21 | 2017-02-14 | Monsanto Technology Llc | Soybean variety XR47AF15X |
US9609831B1 (en) | 2016-03-22 | 2017-04-04 | Monsanto Technology Llc | Soybean cultivar 56052100 |
US9609832B1 (en) | 2016-03-22 | 2017-04-04 | Monsanto Technology Llc | Soybean cultivar 53032202 |
US9585357B1 (en) | 2016-03-22 | 2017-03-07 | Monsanto Technology Llc | Soybean cultivar 53146341 |
US9585356B1 (en) | 2016-03-22 | 2017-03-07 | Monsanto Technology Llc | Soybean cultivar 52232331 |
US9648836B1 (en) | 2016-03-22 | 2017-05-16 | Monsanto Technology Llc | Soybean cultivar 50262411 |
US9706729B1 (en) | 2016-03-22 | 2017-07-18 | Monsanto Technology Llc | Soybean cultivar 51240902 |
US9591827B1 (en) | 2016-03-22 | 2017-03-14 | Monsanto Technology Llc | Soybean cultivar 57160653 |
US9578849B1 (en) | 2016-03-22 | 2017-02-28 | Monsanto Technology Llc | Soybean cultivar 55021012 |
US9648835B1 (en) | 2016-03-22 | 2017-05-16 | Monsanto Technology Llc | Soybean cultivar 53030521 |
US9609834B1 (en) | 2016-03-22 | 2017-04-04 | Monsanto Technology Llc | Soybean cultivar 55030014 |
US9609833B1 (en) | 2016-03-22 | 2017-04-04 | Monsanto Technology Llc | Soybean cultivar 58111502 |
US9648834B1 (en) | 2016-03-22 | 2017-05-16 | Monsanto Technology Llc | Soybean cultivar 51201110 |
US9770000B1 (en) | 2016-03-22 | 2017-09-26 | Monsanto Technology Llc | Soybean cultivar 56042428 |
US9655336B1 (en) | 2016-03-22 | 2017-05-23 | Monsanto Technology Llc | Soybean cultivar 58150703 |
US9756823B1 (en) | 2016-05-31 | 2017-09-12 | Monsanto Technology Llc | Soybean variety 01056961 |
US9867352B2 (en) | 2016-05-31 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01057081 |
US9713311B1 (en) | 2016-05-31 | 2017-07-25 | Monsanto Technology Llc | Soybean variety 01057181 |
US9750218B1 (en) | 2016-05-31 | 2017-09-05 | Monsanto Technology Llc | Soybean variety 01056968 |
US9888648B2 (en) | 2016-05-31 | 2018-02-13 | Monsanto Technology Llc | Soybean variety 01057115 |
US9907245B2 (en) | 2016-05-31 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01056902 |
US9888646B2 (en) | 2016-05-31 | 2018-02-13 | Monsanto Technology Llc | Soybean variety 01057021 |
US9924672B2 (en) | 2016-05-31 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01056984 |
US9901052B2 (en) | 2016-05-31 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01057176 |
US9907247B2 (en) | 2016-05-31 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01056929 |
US10226001B2 (en) | 2016-05-31 | 2019-03-12 | Monsanto Technology Llc | Soybean variety 01056938 |
US10051806B2 (en) | 2016-05-31 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01057079 |
US9743623B1 (en) | 2016-05-31 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01056977 |
US9907246B2 (en) | 2016-05-31 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01056904 |
US9901051B2 (en) | 2016-05-31 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01057075 |
US9867351B2 (en) | 2016-05-31 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01057076 |
US9877450B2 (en) | 2016-05-31 | 2018-01-30 | Monsanto Technology Llc | Soybean variety 01056979 |
US9872469B2 (en) | 2016-05-31 | 2018-01-23 | Monsanto Technology Llc | Soybean variety 01056991 |
US9888647B2 (en) | 2016-05-31 | 2018-02-13 | Monsanto Technology Llc | Soybean variety 01057072 |
US9743624B1 (en) | 2016-05-31 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01056994 |
US9788515B1 (en) | 2016-06-01 | 2017-10-17 | Monsanto Technology Llc | Soybean variety 01057347 |
US9907252B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058320 |
US10104847B2 (en) | 2016-06-01 | 2018-10-23 | Monsanto Technology Llc | Soybean variety 01057454 |
US9907249B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057459 |
US9706738B1 (en) | 2016-06-01 | 2017-07-18 | Monsanto Technology Llc | Soybean variety 01057282 |
US9848562B1 (en) | 2016-06-01 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01058349 |
US10226002B2 (en) | 2016-06-01 | 2019-03-12 | Monsanto Technology Llc | Soybean variety 01057408 |
US10231398B2 (en) | 2016-06-01 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01057400 |
US9700008B1 (en) | 2016-06-01 | 2017-07-11 | Monsanto Technology Llc | Soybean variety 01057341 |
US9854772B2 (en) | 2016-06-01 | 2018-01-02 | Monsanto Technology Llc | Soybean variety 01058360 |
US9968054B2 (en) | 2016-06-01 | 2018-05-15 | Monsanto Technology Llc | Soybean variety 01057207 |
US9848563B1 (en) | 2016-06-01 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01058353 |
US9832953B1 (en) | 2016-06-01 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01057348 |
US9788514B1 (en) | 2016-06-01 | 2017-10-17 | Monsanto Technology Llc | Soybean variety 01057345 |
US9661823B1 (en) | 2016-06-01 | 2017-05-30 | Monsanto Technology Llc | Soybean variety 01057424 |
US9974264B2 (en) | 2016-06-01 | 2018-05-22 | Monsanto Technology Llc | Soybean variety 01057212 |
US9907250B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057493 |
US9706739B1 (en) | 2016-06-01 | 2017-07-18 | Monsanto Technology Llc | Soybean variety 01057290 |
US9907248B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01057390 |
US9907251B2 (en) | 2016-06-01 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058313 |
US9907254B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058698 |
US9693530B1 (en) | 2016-06-02 | 2017-07-04 | Monsanto Technology Llc | Soybean variety 01063884 |
US9872470B2 (en) | 2016-06-02 | 2018-01-23 | Monsanto Technology Llc | Soybean variety 01058469 |
US9907255B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058767 |
US9832954B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01058711 |
US9807960B1 (en) | 2016-06-02 | 2017-11-07 | Monsanto Technology Llc | Soybean variety 01063888 |
US9907256B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058822 |
US9854773B2 (en) | 2016-06-02 | 2018-01-02 | Monsanto Technology Llc | Soybean variety 01058444 |
US9700009B1 (en) | 2016-06-02 | 2017-07-11 | Monsanto Technology Llc | Soybean variety 01058861 |
US9867354B2 (en) | 2016-06-02 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01058529 |
US9832956B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01063886 |
US9700010B1 (en) | 2016-06-02 | 2017-07-11 | Monsanto Technology Llc | Soybean variety 01063871 |
US9907253B2 (en) | 2016-06-02 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01058513 |
US9832957B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01058863 |
US9867353B2 (en) | 2016-06-02 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01058528 |
US9901053B2 (en) | 2016-06-02 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01058606 |
US9693529B1 (en) | 2016-06-02 | 2017-07-04 | Monsanto Technology Llc | Soybean variety 01063878 |
US9848564B1 (en) | 2016-06-02 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01058864 |
US9832955B1 (en) | 2016-06-02 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01058718 |
US10219462B2 (en) | 2016-06-03 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01063907 |
US9717211B1 (en) | 2016-06-03 | 2017-08-01 | Monsanto Technology Llc | Soybean variety 01063940 |
US9907258B2 (en) | 2016-06-03 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01063898 |
US9848565B1 (en) | 2016-06-03 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01063939 |
US9730419B1 (en) | 2016-06-03 | 2017-08-15 | Monsanto Technology Llc | Soybean variety 01064108 |
US10058047B2 (en) | 2016-06-03 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01063918 |
US9717212B1 (en) | 2016-06-03 | 2017-08-01 | Monsanto Technology Llc | Soybean variety 01064113 |
US9907259B2 (en) | 2016-06-03 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01063916 |
US9717213B1 (en) | 2016-06-03 | 2017-08-01 | Monsanto Technology Llc | Soybean variety 01064120 |
US9750221B1 (en) | 2016-06-03 | 2017-09-05 | Monsanto Technology Llc | Soybean variety 01063935 |
US9730418B1 (en) | 2016-06-03 | 2017-08-15 | Monsanto Technology Llc | Soybean variety 01063920 |
US9737037B1 (en) | 2016-06-03 | 2017-08-22 | Monsanto Technology Llc | Soybean variety 01063921 |
US9832958B1 (en) | 2016-06-03 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01064122 |
US9907261B2 (en) | 2016-06-03 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01063928 |
US9907260B2 (en) | 2016-06-03 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01063906 |
US9955645B2 (en) | 2016-06-03 | 2018-05-01 | Monsanto Technology Llc | Soybean variety 01063902 |
US9901054B2 (en) | 2016-06-03 | 2018-02-27 | Monsanto Technology Llc | Soybean variety 01063900 |
US9743625B1 (en) | 2016-06-03 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01063903 |
US9907257B2 (en) | 2016-06-03 | 2018-03-06 | Monsanto Technology Llc | Soybean variety 01063892 |
US9832959B1 (en) | 2016-06-06 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01064131 |
US9743627B1 (en) | 2016-06-06 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01064182 |
US9832961B1 (en) | 2016-06-06 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01064167 |
US9867356B2 (en) | 2016-06-06 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01064143 |
US9743626B1 (en) | 2016-06-06 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01064148 |
US9872471B2 (en) | 2016-06-06 | 2018-01-23 | Monsanto Technology Llc | Soybean variety 01064177 |
US9867355B2 (en) | 2016-06-06 | 2018-01-16 | Monsanto Technology Llc | Soybean variety 01064176 |
US9848566B1 (en) | 2016-06-06 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01064124 |
US9832960B1 (en) | 2016-06-06 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01064152 |
US9795106B1 (en) | 2016-06-06 | 2017-10-24 | Monsanto Technology Llc | Soybean variety 01064150 |
US10064356B2 (en) | 2016-06-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064137 |
US9839187B1 (en) | 2016-06-06 | 2017-12-12 | Monsanto Technology Llc | Soybean variety 01064165 |
US9730420B1 (en) | 2016-06-06 | 2017-08-15 | Monsanto Technology Llc | Soybean variety 01064123 |
US9844197B1 (en) | 2016-06-06 | 2017-12-19 | Monsanto Technology Llc | Soybean variety 01064151 |
US9924673B2 (en) | 2016-06-06 | 2018-03-27 | Monsanto Technology Llc | Soybean variety 01064169 |
US9848567B1 (en) | 2016-06-06 | 2017-12-26 | Monsanto Technology Llc | Soybean variety 01064142 |
US9832962B1 (en) | 2016-06-06 | 2017-12-05 | Monsanto Technology Llc | Soybean variety 01064172 |
US9743629B1 (en) | 2016-06-07 | 2017-08-29 | Monsanto Technology Llc | Soybean variety 01058714 |
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 |
US9907281B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 64291310 |
US9930858B1 (en) | 2017-03-29 | 2018-04-03 | Monsanto Technology Llc | Soybean cultivar 62232543 |
US9913448B1 (en) | 2017-03-29 | 2018-03-13 | Monsanto Technology Llc | Soybean cultivar 63281310 |
US9999193B1 (en) | 2017-03-29 | 2018-06-19 | Monsanto Technology Llc | Soybean cultivar 65090203 |
US10039249B1 (en) | 2017-03-29 | 2018-08-07 | Monsanto Technology Llc | Soybean cultivar 61162227 |
US9913449B1 (en) | 2017-03-29 | 2018-03-13 | Monsanto Technology Llc | Soybean cultivar 67272133 |
US9913450B1 (en) | 2017-03-29 | 2018-03-13 | Monsanto Technology Llc | Soybean cultivar 68071303 |
US9930856B1 (en) | 2017-03-29 | 2018-04-03 | Monsanto Technology Llc | Soybean cultivar 62030513 |
US9907283B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 62103227 |
US9907279B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 63282422 |
US9907282B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 60371632 |
US9907280B1 (en) | 2017-03-29 | 2018-03-06 | Monsanto Technology Llc | Soybean cultivar 64492916 |
US9888659B1 (en) | 2017-03-29 | 2018-02-13 | Monsanto Technology Llc | Soybean cultivar 61040705 |
US9961868B1 (en) | 2017-03-29 | 2018-05-08 | Monsanto Technology Llc | Soybean cultivar 60111034 |
US9930857B1 (en) | 2017-03-29 | 2018-04-03 | Monsato Technology Llc | Soybean cultivar 61311131 |
US9894869B1 (en) | 2017-03-29 | 2018-02-20 | Monsanto Technology Llc | Soybean cultivar 64091728 |
US10034447B1 (en) | 2017-06-26 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064421 |
US10034448B1 (en) | 2017-06-26 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064516 |
US10314273B2 (en) | 2017-06-26 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01068110 |
US10342199B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01068883 |
US10405511B2 (en) | 2017-06-26 | 2019-09-10 | Monsanto Technology Llc | Soybean variety 01064350 |
US10342197B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01064349 |
US10039254B1 (en) | 2017-06-26 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064117 |
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 |
US10383304B2 (en) | 2017-06-26 | 2019-08-20 | Monsanto Technology Llc | Soybean variety 01063944 |
US10368518B2 (en) | 2017-06-26 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01068117 |
US10039255B1 (en) | 2017-06-26 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064252 |
US10342198B2 (en) | 2017-06-26 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01064381 |
US10010046B1 (en) | 2017-06-26 | 2018-07-03 | Monsanto Technology Llc | Soybean variety 01064401 |
US10053699B1 (en) | 2017-06-28 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01068892 |
US10178837B1 (en) | 2017-06-28 | 2019-01-15 | Monsanto Technology Llc | Soybean variety 5PHQP31 |
US10349598B2 (en) | 2017-06-28 | 2019-07-16 | Monsanto Technology Llc | Soybean variety 01068088 |
US10172310B1 (en) | 2017-06-28 | 2019-01-08 | Monsanto Technology Llc | Soybean variety 5PYFS21 |
US10178838B1 (en) | 2017-06-28 | 2019-01-15 | Monsanto Technology Llc | Soybean variety 5PQNG18 |
US10244705B2 (en) | 2017-06-28 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064265 |
US10182539B1 (en) | 2017-06-28 | 2019-01-22 | Monsanto Technology Llc | Soybean variety 5PDZB74 |
US10194612B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01068093 |
US10206350B1 (en) | 2017-06-28 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 5PDSD70 |
US10194613B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064363 |
US10036027B1 (en) | 2017-06-28 | 2018-07-31 | Monsanto Technology Llc | Soybean variety 01064536 |
US10368512B2 (en) | 2017-06-28 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 01068083 |
US10244704B2 (en) | 2017-06-28 | 2019-04-02 | Monsanto Technologies Llc | Soybean variety 01068084 |
US10011845B1 (en) | 2017-06-28 | 2018-07-03 | Monsanto Technology Llc | Soybean variety 01068893 |
US10196646B2 (en) | 2017-06-28 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064344 |
US10292358B2 (en) | 2017-06-28 | 2019-05-21 | Monsanto Technology Llc | Soybean variety 01064327 |
US10206349B1 (en) | 2017-06-28 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 5PPHU37 |
US10201138B2 (en) | 2017-06-28 | 2019-02-12 | Monsanto Technology Llc | Soybean variety 01064519 |
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 |
US10271493B2 (en) | 2017-06-28 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064280 |
US10188059B1 (en) | 2017-06-28 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 5PUWT62 |
US10201139B2 (en) | 2017-07-06 | 2019-02-12 | Monsanto Technology Llc | Soybean variety 01068144 |
US10098305B1 (en) | 2017-07-06 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01064403 |
US10194614B2 (en) | 2017-07-06 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064567 |
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 |
US10064379B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064450 |
US10194616B2 (en) | 2017-07-06 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064568 |
US10231403B2 (en) | 2017-07-06 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068141 |
US10064381B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01067510 |
US10070616B1 (en) | 2017-07-06 | 2018-09-11 | Monsanto Technology Llc | Soybean variety 01067507 |
US10244706B2 (en) | 2017-07-06 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064571 |
US10064367B1 (en) | 2017-07-06 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01067515 |
US10238057B2 (en) | 2017-07-06 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064461 |
US10136602B1 (en) | 2017-07-19 | 2018-11-27 | Monsanto Technology Llc | Soybean variety 01068087 |
US10219466B2 (en) | 2017-07-19 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064243 |
US10426116B2 (en) | 2017-07-19 | 2019-10-01 | Monsanto Technology Llc | Soybean variety 01068124 |
US10238076B2 (en) | 2017-07-19 | 2019-03-26 | Monsanto Technology, Llc | Soybean variety 01068130 |
US10231422B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068134 |
US10440908B2 (en) | 2017-07-19 | 2019-10-15 | Monsanto Technology Llc | Soybean variety 01064245 |
US10212901B2 (en) | 2017-07-19 | 2019-02-26 | Monsanto Technology Llc | Soybean variety 01068895 |
US10231405B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068135 |
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 |
US10064368B1 (en) | 2017-07-19 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01068133 |
US10064369B1 (en) | 2017-07-19 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01068131 |
US10098306B1 (en) | 2017-07-19 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01068070 |
US10231404B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064187 |
US10194617B1 (en) | 2017-07-19 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01068066 |
US10398105B2 (en) | 2017-07-19 | 2019-09-03 | Monsanto Technology Llc | Soybean variety 01064198 |
US10231421B2 (en) | 2017-07-19 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064231 |
US10058060B1 (en) | 2017-07-19 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01068082 |
US10314275B2 (en) | 2017-07-19 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064238 |
US10058061B1 (en) | 2017-07-19 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01068112 |
US10206352B2 (en) | 2017-07-19 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 01068065 |
US10098307B1 (en) | 2017-07-20 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01064838 |
US10231406B2 (en) | 2017-07-20 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01068831 |
US10117400B1 (en) | 2017-07-20 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064185 |
US10058062B1 (en) | 2017-07-20 | 2018-08-28 | Monsanto Technology Llc | Soybean variety 01064840 |
US10117401B1 (en) | 2017-07-20 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064488 |
US10285367B2 (en) | 2017-07-20 | 2019-05-14 | Monsanto Technology Llc | Soybean variety 01064508 |
US10244723B2 (en) | 2017-07-20 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01068058 |
US10143167B1 (en) | 2017-07-20 | 2018-12-04 | Monsanto Technology Llc | Soybean variety 01064871 |
US10098308B1 (en) | 2017-07-20 | 2018-10-16 | Monsanto Technology Llc | Soybean variety 01068054 |
US10064370B1 (en) | 2017-07-20 | 2018-09-04 | Monsanto Technology Llc | Soybean variety 01064806 |
US10051823B1 (en) | 2017-07-20 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064868 |
US10206353B2 (en) | 2017-07-20 | 2019-02-19 | Monsanto Technology Llc | Soybean variety 01064485 |
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 |
US10244708B2 (en) | 2017-08-23 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064594 |
US10238059B2 (en) | 2017-08-23 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064460 |
US10117403B1 (en) | 2017-08-23 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064099 |
US10264748B2 (en) | 2017-08-23 | 2019-04-23 | Monsanto Technology Llc | Soybean variety 01064212 |
US10251354B2 (en) | 2017-08-23 | 2019-04-09 | Monsanto Technology Llc | Soybean variety 01064596 |
US10257994B2 (en) | 2017-08-23 | 2019-04-16 | Monsanto Technology Llc | Soybean variety 01068143 |
US10117404B1 (en) | 2017-08-23 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064613 |
US10231408B1 (en) | 2017-08-23 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064013 |
US10285368B2 (en) | 2017-08-23 | 2019-05-14 | Monsanto Technology Llc | Soybean variety 01064056 |
US10231407B1 (en) | 2017-08-23 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01063959 |
US10271494B2 (en) | 2017-08-23 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01068122 |
US10045506B1 (en) | 2017-08-24 | 2018-08-14 | Monsanto Technology Llc | Soybean variety 01064415 |
US10039256B1 (en) | 2017-08-24 | 2018-08-07 | Monsanto Technology Llc | Soybean variety 01064446 |
US10231423B1 (en) | 2017-08-24 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064404 |
US10051824B1 (en) | 2017-08-24 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064393 |
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 |
US10136603B1 (en) | 2017-08-24 | 2018-11-27 | Monsanto Technology Llc | Soybean variety 01067535 |
US10130058B1 (en) | 2017-08-24 | 2018-11-20 | Monsanto Technology Llc | Soybean variety 01064723 |
US10314276B2 (en) | 2017-08-24 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064370 |
US10051825B1 (en) | 2017-08-24 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064400 |
US10130060B1 (en) | 2017-08-24 | 2018-11-20 | Monsanto Technology Llc | Soybean variety 01068804 |
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 |
US10271496B2 (en) | 2017-08-29 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064102 |
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 |
US10314265B2 (en) | 2017-08-29 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064669 |
US10271495B2 (en) | 2017-08-29 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064036 |
US10159205B1 (en) | 2017-08-29 | 2018-12-25 | Monsanto Technology Llc | Soybean variety 01064633 |
US10342201B2 (en) | 2017-08-29 | 2019-07-09 | Monsanto Technology Llc | Soybean variety 01067531 |
US10231409B1 (en) | 2017-08-29 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064026 |
US10244724B2 (en) | 2017-08-29 | 2019-04-02 | Monsanto Technology Llc | Soybean variety 01064667 |
US10238077B2 (en) | 2017-08-29 | 2019-03-26 | Monsanto Technology Llc | Soybean variety 01064104 |
US10159206B1 (en) | 2017-08-29 | 2018-12-25 | Monsanto Technology Llc | Soybean variety 01067519 |
US10257995B2 (en) | 2017-08-29 | 2019-04-16 | Monsanto Technology Llc | Soybean variety 01064083 |
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 |
US10271498B2 (en) | 2017-08-30 | 2019-04-30 | Monsanto Technology Llc | Soybean variety 01064676 |
US10357007B2 (en) | 2017-08-30 | 2019-07-23 | Monsanto Technology Llc | Soybean variety 01064743 |
US10117406B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064166 |
US10117405B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064511 |
US10219467B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064677 |
US10314278B2 (en) | 2017-08-30 | 2019-06-11 | Monsanto Technology Llc | Soybean variety 01064635 |
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 |
US10231424B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01067526 |
US10117409B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064756 |
US10051826B1 (en) | 2017-08-30 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064189 |
US10231425B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01067534 |
US10219485B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01069661 |
US10188072B1 (en) | 2017-08-30 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 01064694 |
US10194618B1 (en) | 2017-08-30 | 2019-02-05 | Monsanto Technology Llc | Soybean variety 01064184 |
US10231426B1 (en) | 2017-08-30 | 2019-03-19 | Monsanto Technology Llc | Soybean variety 01064748 |
US10278350B2 (en) | 2017-08-30 | 2019-05-07 | Monsanto Technology Llc | Soybean variety 01064672 |
US10212902B1 (en) | 2017-08-30 | 2019-02-26 | Monsanto Technology Llc | Soybean variety 01067523 |
US10117407B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064490 |
US10219484B1 (en) | 2017-08-30 | 2019-03-05 | Monsanto Technology Llc | Soybean variety 01064735 |
US10051827B1 (en) | 2017-08-30 | 2018-08-21 | Monsanto Technology Llc | Soybean variety 01064164 |
US10188061B1 (en) | 2017-08-30 | 2019-01-29 | Monsanto Technology Llc | Soybean variety 01064486 |
US10117408B1 (en) | 2017-08-30 | 2018-11-06 | Monsanto Technology Llc | Soybean variety 01064491 |
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 |
US10368520B1 (en) | 2018-05-24 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 5PCDJ10 |
US10477818B1 (en) | 2018-05-24 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 5PTJB40 |
US10368519B1 (en) | 2018-05-24 | 2019-08-06 | Monsanto Technology Llc | Soybean variety 5PYGS18 |
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 |
US10517255B1 (en) | 2018-08-06 | 2019-12-31 | Monsanto Technology Llc | Soybean cultivar 62391708 |
US10342204B1 (en) | 2018-08-06 | 2019-07-09 | Monsanto Technology Llc | Soybean cultivar 65382631 |
US10398122B1 (en) | 2018-08-06 | 2019-09-03 | Monsanto Technology Llc | Soybean cultivar 77252418 |
US10238079B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 77301427 |
US10368521B1 (en) | 2018-08-06 | 2019-08-06 | Monsanto Technology Llc | Soybean cultivar 70490447 |
US10342207B1 (en) | 2018-08-06 | 2019-07-09 | Monsanto Technology Llc | Soybean cultivar 68381333 |
US10238066B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 73301323 |
US10321654B1 (en) | 2018-08-06 | 2019-06-18 | Monsanto Technology Llc | Soybean cultivar 76222418 |
US10357010B1 (en) | 2018-08-06 | 2019-07-23 | Monsanto Technology Llc | Soybean cultivar 68352516 |
US10258014B1 (en) | 2018-08-06 | 2019-04-16 | Monsanto Technology Llc | Soybean cultivar 76051047 |
US10264760B1 (en) | 2018-08-06 | 2019-04-23 | Monsanto Technology Llc | Soybean cultivar 79380729 |
US10238080B1 (en) | 2018-08-06 | 2019-03-26 | Monsanto Technology Llc | Soybean cultivar 79072143 |
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 |
US10349607B1 (en) | 2018-08-06 | 2019-07-16 | Monsanto Technology Llc | Soybean cultivar 78091047 |
US10349608B1 (en) | 2018-08-06 | 2019-07-16 | Monsanto Technology Llc | Soybean cultivar 70520427 |
US10258013B1 (en) | 2018-08-06 | 2019-04-16 | Monsanto Technology Llc | Soybean cultivar 64442216 |
US10993404B2 (en) | 2018-08-22 | 2021-05-04 | Monsanto Technology Llc | Soybean variety 01072356 |
US11096363B2 (en) | 2018-08-22 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01072331 |
US11109553B2 (en) | 2018-08-22 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01072328 |
US10561098B1 (en) | 2018-08-22 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01067649 |
US10624285B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01068376 |
US10582686B1 (en) | 2018-08-22 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01064580 |
US11026389B2 (en) | 2018-08-22 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01072367 |
US10555484B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068903 |
US10918065B2 (en) | 2018-08-22 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072343 |
US10897869B2 (en) | 2018-08-22 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01072333 |
US10555482B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068427 |
US10575489B1 (en) | 2018-08-22 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01067545 |
US10973200B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072342 |
US10624303B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072279 |
US10999998B2 (en) | 2018-08-22 | 2021-05-11 | Monsanto Technology Llc | Soybean variety 01072346 |
US10624302B2 (en) | 2018-08-22 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072280 |
US10939652B2 (en) | 2018-08-22 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072350 |
US11013197B2 (en) | 2018-08-22 | 2021-05-25 | Monsanto Technology Llc | Soybean variety 01072368 |
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 |
US10973198B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072340 |
US10555481B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068364 |
US10897870B2 (en) | 2018-08-22 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01072322 |
US10555483B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067598 |
US10555480B1 (en) | 2018-08-22 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068350 |
US10973201B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072339 |
US11206791B2 (en) | 2018-08-22 | 2021-12-28 | Monsanto Technology Llc | Soybean variety 01072337 |
US10905081B2 (en) | 2018-08-22 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072329 |
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 |
US11044876B2 (en) | 2018-08-22 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01072345 |
US11109552B2 (en) | 2018-08-22 | 2021-09-07 | Monsanto Technology Llc | Soybean variety 01072332 |
US10905080B2 (en) | 2018-08-22 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072355 |
US10602704B2 (en) | 2018-08-22 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068313 |
US10617089B2 (en) | 2018-08-22 | 2020-04-14 | Monsanto Technology Llc | Soybean variety 01067827 |
US10973199B2 (en) | 2018-08-22 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072341 |
US10512239B1 (en) | 2018-08-23 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072273 |
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 |
US10506786B1 (en) | 2018-08-23 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01067789 |
US10687501B2 (en) | 2018-08-23 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068438 |
US10555485B1 (en) | 2018-08-23 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01073532 |
US10499601B1 (en) | 2018-08-23 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01068356 |
US10463000B1 (en) | 2018-08-23 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01072224 |
US10631486B2 (en) | 2018-08-23 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068187 |
US10624287B2 (en) | 2018-08-23 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072275 |
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 |
US10524446B1 (en) | 2018-08-23 | 2020-01-07 | Monsanto Technology Llc | Soybean variety 01072245 |
US10687502B2 (en) | 2018-08-23 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068433 |
US10638707B2 (en) | 2018-08-23 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01067639 |
US10609888B2 (en) | 2018-08-23 | 2020-04-07 | Monsanto Technology Llc | Soybean variety 01068346 |
US10624286B2 (en) | 2018-08-23 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01072276 |
US10638708B2 (en) | 2018-08-23 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068325 |
US10492450B1 (en) | 2018-08-23 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01064752 |
US10548284B1 (en) | 2018-08-23 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01072294 |
US10660292B2 (en) | 2018-08-24 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01068157 |
US10448610B1 (en) | 2018-08-24 | 2019-10-22 | Monsanto Technology Llc | Soybean variety 01067781 |
US10631504B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01072295 |
US10638709B2 (en) | 2018-08-24 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068191 |
US10575491B1 (en) | 2018-08-24 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01072289 |
US10506787B1 (en) | 2018-08-24 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01072220 |
US10548285B1 (en) | 2018-08-24 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01067703 |
US10477821B1 (en) | 2018-08-24 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01067769 |
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 |
US10561104B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01072234 |
US10561102B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068182 |
US10531629B1 (en) | 2018-08-24 | 2020-01-14 | Monsanto Technology Llc | Soybean variety 01072223 |
US10561100B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01067742 |
US10582689B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067573 |
US10638710B2 (en) | 2018-08-24 | 2020-05-05 | Monsanto Technology Llc | Soybean variety 01068195 |
US10631500B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067788 |
US10512241B1 (en) | 2018-08-24 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072232 |
US10561103B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01073501 |
US10561106B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068466 |
US10561105B1 (en) | 2018-08-24 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01072235 |
US10485213B1 (en) | 2018-08-24 | 2019-11-26 | Monsanto Technology Llc | Soybean variety 01068968 |
US10512240B1 (en) | 2018-08-24 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01072228 |
US10542721B1 (en) | 2018-08-24 | 2020-01-28 | Monsanto Technology Llc | Soybean variety 01072231 |
US10555487B1 (en) | 2018-08-24 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067694 |
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 |
US10470425B1 (en) | 2018-08-24 | 2019-11-12 | Monsanto Technology Llc | Soybean variety 01067783 |
US10624304B2 (en) | 2018-08-24 | 2020-04-21 | Monsanto Technology Llc | Soybean variety 01068381 |
US10517259B1 (en) | 2018-08-24 | 2019-12-31 | Monsanto Technology Llc | Soybean variety 01068824 |
US10555488B1 (en) | 2018-08-24 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01067714 |
US10463010B1 (en) | 2018-08-24 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01068189 |
US10617090B2 (en) | 2018-08-24 | 2020-04-14 | Monsanto Technology Llc | Soybean variety 01068163 |
US10631507B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068485 |
US10492451B1 (en) | 2018-08-24 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 0107226 |
US10470395B1 (en) | 2018-08-24 | 2019-11-12 | Monsanto Technology Llc | Soybean variety 01067663 |
US10631503B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067814 |
US10582687B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01068359 |
US10645898B2 (en) | 2018-08-24 | 2020-05-12 | Monsanto Technology Llc | Soybean variety 01068192 |
US10631501B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067793 |
US10582688B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067543 |
US10631506B2 (en) | 2018-08-24 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068838 |
US10582690B1 (en) | 2018-08-24 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01067790 |
US10463013B1 (en) | 2018-08-27 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01069709 |
US10582678B1 (en) | 2018-08-27 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01072239 |
US10568291B1 (en) | 2018-08-27 | 2020-02-25 | Monsanto Technology Llc | Soybean variety 01067741 |
US10582679B1 (en) | 2018-08-27 | 2020-03-10 | Monsanto Technology Llc | Soybean variety 01072243 |
US10602691B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01072253 |
US10568290B1 (en) | 2018-08-27 | 2020-02-25 | Monsanto Technology Llc | Soybean variety 01068219 |
US10477823B1 (en) | 2018-08-27 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01069720 |
US10499602B1 (en) | 2018-08-27 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01067553 |
US10561107B1 (en) | 2018-08-27 | 2020-02-18 | Monsanto Technology Llc | Soybean variety 01068807 |
US10602706B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01077437 |
US10660293B2 (en) | 2018-08-27 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01077434 |
US10555489B1 (en) | 2018-08-27 | 2020-02-11 | Monsanto Technology Llc | Soybean variety 01068972 |
US10602690B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01068227 |
US10660294B2 (en) | 2018-08-27 | 2020-05-26 | Monsanto Technology Llc | Soybean variety 01077435 |
US10512242B1 (en) | 2018-08-27 | 2019-12-24 | Monsanto Technology Llc | Soybean variety 01077436 |
US10687503B2 (en) | 2018-08-27 | 2020-06-23 | Monsanto Technology Llc | Soybean variety 01068213 |
US10477822B1 (en) | 2018-08-27 | 2019-11-19 | Monsanto Technology Llc | Soybean variety 01069686 |
US10631508B2 (en) | 2018-08-27 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01068228 |
US10492453B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01072254 |
US10492452B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01067646 |
US10548286B1 (en) | 2018-08-27 | 2020-02-04 | Monsanto Technology Llc | Soybean variety 01068171 |
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 |
US10743493B2 (en) | 2018-08-27 | 2020-08-18 | Monsanto Technology Llc | Soybean variety 01077438 |
US10492454B1 (en) | 2018-08-27 | 2019-12-03 | Monsanto Technology Llc | Soybean variety 01072256 |
US10631509B2 (en) | 2018-08-27 | 2020-04-28 | Monsanto Technology Llc | Soybean variety 01067807 |
US10575492B1 (en) | 2018-08-27 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01068915 |
US10463012B1 (en) | 2018-08-27 | 2019-11-05 | Monsanto Technology Llc | Soybean variety 01069688 |
US10743492B2 (en) | 2018-08-27 | 2020-08-18 | Monsanto Technology Llc | Soybean variety 01077433 |
US10506788B1 (en) | 2018-08-27 | 2019-12-17 | Monsanto Technology Llc | Soybean variety 01072292 |
US10602692B2 (en) | 2018-08-27 | 2020-03-31 | Monsanto Technology Llc | Soybean variety 01072252 |
US10575493B1 (en) | 2018-08-27 | 2020-03-03 | Monsanto Technology Llc | Soybean variety 01067819 |
US10499603B1 (en) | 2018-08-27 | 2019-12-10 | Monsanto Technology Llc | Soybean variety 01068207 |
US10506789B1 (en) | 2018-12-04 | 2019-12-17 | Monsanto Technology Llc | Soybean cultivar 61381330 |
US10617080B1 (en) | 2019-06-04 | 2020-04-14 | Monsanto Technology Llc | Soybean cultivar 87352527 |
US10631513B1 (en) | 2019-06-04 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 87181312 |
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 |
US10631512B1 (en) | 2019-06-04 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 85322536 |
US10602693B1 (en) | 2019-06-04 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 81302732 |
US10667484B1 (en) | 2019-06-04 | 2020-06-02 | Monsanto Technology Llc | Soybean cultivar 82322805 |
US10609883B1 (en) | 2019-06-05 | 2020-04-07 | Monsanto Technology Llc | Soybean cultivar 82340103 |
US10602708B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 82370721 |
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 |
US10660295B1 (en) | 2019-06-05 | 2020-05-26 | Monsanto Technology Llc | Soybean cultivar 86072029 |
US10638686B1 (en) | 2019-06-05 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 84161023 |
US10602707B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 85442619 |
US10624288B1 (en) | 2019-06-05 | 2020-04-21 | Monsanto Technology Llc | Soybean cultivar 85061635 |
US10631514B1 (en) | 2019-06-05 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 80131402 |
US10602709B1 (en) | 2019-06-05 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 86360847 |
US10638714B1 (en) | 2019-06-06 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 84062517 |
US10548289B1 (en) | 2019-06-06 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 82290447 |
US10568292B1 (en) | 2019-06-06 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 80181322 |
US10631487B1 (en) | 2019-06-06 | 2020-04-28 | Monsanto Technology Llc | Soybean cultivar 83240828 |
US10561108B1 (en) | 2019-06-06 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 80431932 |
US10617092B1 (en) | 2019-06-06 | 2020-04-14 | Monsanto Technology Llc | Soybean cultivar 80411617 |
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 |
US10813335B1 (en) | 2019-06-06 | 2020-10-27 | Monsanto Technology Llc | Soybean cultivar 80261309 |
US10609889B1 (en) | 2019-06-06 | 2020-04-07 | Monsanto Technology Llc | Soybean cultivar 89001711 |
US10638687B1 (en) | 2019-06-06 | 2020-05-05 | Monsanto Technology Llc | Soybean cultivar 88103087 |
US10561109B1 (en) | 2019-06-06 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 82042815 |
US10785950B1 (en) | 2019-06-06 | 2020-09-29 | Stine Seed Farm, Inc. | Soybean cultivar 86190345 |
US10568284B1 (en) | 2019-06-06 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 87161804 |
US10555491B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 83072718 |
US10568294B1 (en) | 2019-06-07 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar 82440038 |
US10555492B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 81430038 |
US10561090B1 (en) | 2019-06-07 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 86370345 |
US10602710B1 (en) | 2019-06-07 | 2020-03-31 | Monsanto Technology Llc | Soybean cultivar 89282206 |
US10561089B1 (en) | 2019-06-07 | 2020-02-18 | Monsanto Technology Llc | Soybean cultivar 84060345 |
US10548290B1 (en) | 2019-06-07 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 89031941 |
US10568293B1 (en) | 2019-06-07 | 2020-02-25 | Monsanto Technology Llc | Soybean cultivar AR 86112649 |
US10555490B1 (en) | 2019-06-07 | 2020-02-11 | Monsanto Technology Llc | Soybean cultivar 85370909 |
US10548274B1 (en) | 2019-06-07 | 2020-02-04 | Monsanto Technology Llc | Soybean cultivar 83262718 |
US10660296B1 (en) | 2019-06-07 | 2020-05-26 | Monsanto Technology Llc | Soybean cultivar 84241941 |
US10918052B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073365 |
US10849300B1 (en) | 2019-07-29 | 2020-12-01 | Monsanto Technology Llc | Soybean variety 01073359 |
US11096357B2 (en) | 2019-07-29 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01073351 |
US10918050B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072791 |
US10925237B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073288 |
US10912270B1 (en) | 2019-07-29 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01072737 |
US10925235B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01072810 |
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 |
US10932429B2 (en) | 2019-07-29 | 2021-03-02 | Monsanto Technology Llc | Soybean variety 01073265 |
US10993401B2 (en) | 2019-07-29 | 2021-05-04 | Monsanto Technology Llc | Soybean variety 01073256 |
US10918051B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01072804 |
US10863709B1 (en) | 2019-07-29 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01072789 |
US10905072B1 (en) | 2019-07-29 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01073330 |
US10925236B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01072823 |
US10863708B1 (en) | 2019-07-29 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01073439 |
US10925238B1 (en) | 2019-07-29 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073320 |
US10932428B2 (en) | 2019-07-29 | 2021-03-02 | Monsanto Technology Llc | Soybean variety 01072749 |
US10918053B1 (en) | 2019-07-29 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073354 |
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 |
US10897864B1 (en) | 2019-07-30 | 2021-01-26 | Monsanto Technology Llc | Soybean variety 01073453 |
US10881074B1 (en) | 2019-07-30 | 2021-01-05 | Monsanto Technology Llc | Soybean variety 01072802 |
US10918055B1 (en) | 2019-07-30 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01073472 |
US10881073B1 (en) | 2019-07-30 | 2021-01-05 | Monsanto Technology Llc | Soybean variety 01072782 |
US10939646B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073479 |
US10888063B1 (en) | 2019-07-30 | 2021-01-12 | Monsanto Technology Llc | Soybean variety 01072800 |
US10925240B1 (en) | 2019-07-30 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073455 |
US11266100B2 (en) | 2019-07-30 | 2022-03-08 | Monsanto Technology Llc | Soybean variety 01073441 |
US10856507B1 (en) | 2019-07-30 | 2020-12-08 | Monsanto Technology Llc | Soybean variety 01073444 |
US10856508B1 (en) | 2019-07-30 | 2020-12-08 | Monsanto Technology Llc | Soybean variety 01073445 |
US10827714B1 (en) | 2019-07-30 | 2020-11-10 | Monsanto Technology Llc | Soybean variety 01073443 |
US10939644B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073461 |
US10939645B2 (en) | 2019-07-30 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01073480 |
US10849301B1 (en) | 2019-07-30 | 2020-12-01 | Monsanto Technology Llc | Soybean variety 01072784 |
US10925239B1 (en) | 2019-07-30 | 2021-02-23 | Monsanto Technology Llc | Soybean variety 01073452 |
US10959389B2 (en) | 2019-07-31 | 2021-03-30 | Monsanto Technology Llc | Soybean variety 01073503 |
US10918056B1 (en) | 2019-07-31 | 2021-02-16 | Monsanto Technology Llc | Soybean variety 01077815 |
US11122759B2 (en) | 2019-07-31 | 2021-09-21 | Monsanto Technology Llc | Soybean variety 01072870 |
US10869453B1 (en) | 2019-07-31 | 2020-12-22 | Monsanto Technology Llc | Soybean variety 01072813 |
US10874078B1 (en) | 2019-07-31 | 2020-12-29 | Monsanto Technology Llc | Soybean variety 01072817 |
US10827715B1 (en) | 2019-07-31 | 2020-11-10 | Monsanto Technology Llc | Soybean variety 01077799 |
US10842116B1 (en) | 2019-07-31 | 2020-11-24 | Monsanto Technology Llc | Soybean variety 01072832 |
US10939647B2 (en) | 2019-07-31 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072860 |
US10939648B2 (en) | 2019-07-31 | 2021-03-09 | Monsanto Technology Llc | Soybean variety 01072899 |
US10863710B1 (en) | 2019-07-31 | 2020-12-15 | Monsanto Technology Llc | Soybean variety 01072819 |
US10842117B1 (en) | 2019-07-31 | 2020-11-24 | Monsanto Technology Llc | Soybean variety 01072842 |
US10973194B2 (en) | 2019-07-31 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072867 |
US10945397B2 (en) | 2019-07-31 | 2021-03-16 | Monsanto Technology Llc | Soybean variety 01073506 |
US10973193B2 (en) | 2019-07-31 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01072864 |
US10912271B1 (en) | 2019-07-31 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01072822 |
US11026387B2 (en) | 2019-08-01 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01073016 |
US11006602B2 (en) | 2019-08-01 | 2021-05-18 | Monsanto Technology Llc | Soybean variety 01072946 |
US10905073B1 (en) | 2019-08-01 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072883 |
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 |
US11019788B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01072980 |
US11019789B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01073534 |
US10905074B1 (en) | 2019-08-01 | 2021-02-02 | Monsanto Technology Llc | Soybean variety 01072906 |
US11019787B2 (en) | 2019-08-01 | 2021-06-01 | Monsanto Technology Llc | Soybean variety 01072951 |
US11129346B2 (en) | 2019-08-01 | 2021-09-28 | Monsanto Technology Llc | Soybean variety 01073533 |
US11026388B2 (en) | 2019-08-01 | 2021-06-08 | Monsanto Technology Llc | Soybean variety 01073060 |
US10912273B1 (en) | 2019-08-07 | 2021-02-09 | Monsanto Technology Llc | Soybean variety 01073355 |
US10980203B2 (en) | 2019-08-07 | 2021-04-20 | Monsanto Technology Llc | Soybean variety 01073305 |
US11096359B2 (en) | 2019-08-07 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01077377 |
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 |
US10973195B2 (en) | 2019-08-07 | 2021-04-13 | Monsanto Technology Llc | Soybean variety 01073325 |
US10952393B2 (en) | 2019-08-15 | 2021-03-23 | Monsanto Technology Llc | Soybean variety 01073169 |
US11013196B2 (en) | 2019-08-15 | 2021-05-25 | Monsanto Technology Llc | Soybean variety 01073094 |
US11044867B2 (en) | 2019-08-15 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073133 |
US11044866B2 (en) | 2019-08-15 | 2021-06-29 | Monsanto Technology Llc | Soybean variety 01073114 |
US10952392B2 (en) | 2019-08-15 | 2021-03-23 | Monsanto Technology Llc | Soybean variety 01073151 |
US11096361B2 (en) | 2019-08-15 | 2021-08-24 | Monsanto Technology Llc | Soybean variety 01073072 |
US10945398B2 (en) | 2019-08-15 | 2021-03-16 | Monsanto Technology Llc | Soybean variety 01073160 |
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2007
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- 2007-10-16 WO PCT/US2007/081527 patent/WO2008048964A2/en active Application Filing
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Cited By (4)
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CN106879622A (zh) * | 2010-05-04 | 2017-06-23 | 陶氏益农公司 | 含有麦草畏衍生物和草甘膦衍生物的协同除草组合物 |
CN112955000A (zh) * | 2018-10-24 | 2021-06-11 | 克莱米特公司 | 使用基于机器学习的种子收获水分预测来改进计算机辅助农场操作 |
CN112955000B (zh) * | 2018-10-24 | 2022-07-19 | 克莱米特有限责任公司 | 使用基于机器学习的种子收获水分预测来改进计算机辅助农场操作 |
CN113416739A (zh) * | 2021-06-24 | 2021-09-21 | 黑龙江八一农垦大学 | 鲁氏酵母菌基因在提高微生物产hdmf的产量中的应用 |
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GT200900064A (es) | 2010-03-04 |
CA2662511C (en) | 2014-12-30 |
WO2008048964A3 (en) | 2008-07-03 |
HN2009000495A (es) | 2014-02-03 |
AU2007311159A1 (en) | 2008-04-24 |
CN101522023B (zh) | 2014-03-12 |
US8754011B2 (en) | 2014-06-17 |
WO2008048964A2 (en) | 2008-04-24 |
ZA200901754B (en) | 2010-03-31 |
EA020590B1 (ru) | 2014-12-30 |
EA200970388A1 (ru) | 2009-08-28 |
AU2007311159B2 (en) | 2012-08-16 |
AR063316A1 (es) | 2009-01-21 |
UA102507C2 (ru) | 2013-07-25 |
US20100279866A1 (en) | 2010-11-04 |
BRPI0717640A2 (pt) | 2013-11-12 |
US20140315717A1 (en) | 2014-10-23 |
EP2053916A2 (en) | 2009-05-06 |
MX2009004115A (es) | 2009-04-30 |
AP2009004826A0 (en) | 2009-04-30 |
CA2662511A1 (en) | 2008-04-24 |
AP2885A (en) | 2014-05-31 |
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