CN102766652A - 编码杀虫蛋白的核苷酸序列 - Google Patents
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- CN102766652A CN102766652A CN2012102575771A CN201210257577A CN102766652A CN 102766652 A CN102766652 A CN 102766652A CN 2012102575771 A CN2012102575771 A CN 2012102575771A CN 201210257577 A CN201210257577 A CN 201210257577A CN 102766652 A CN102766652 A CN 102766652A
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Abstract
本发明涉及编码杀虫蛋白的核苷酸序列。具体地，本发明提供编码表现鳞翅目抑制活性的杀虫蛋白的核苷酸序列，以及在本文中称作Cry1A.105杀虫剂的新杀虫蛋白，表达该杀虫剂的转基因植物，和检测生物样品中该核苷酸序列或杀虫剂的存在的方法。
Description
本申请是申请日为2006年8月30目的中国专利申请200680031606.5“编码杀虫蛋白的核苷酸序列”的分案申请。
技术领域
本发明提供用于植物的新编码序列。该编码序列编码针对广泛鳞翅目物种作物害虫毒性的嵌合杀虫蛋白。
背景技术
天然存在的苏云金芽孢杆菌(B.thuringiensis)分离物的商业制剂已长久用于对农业昆虫害虫的生物控制。根据常规农业实践将从苏云金芽孢杆菌物种发酵获得的Bt孢子和晶体浓缩并配制用于叶施用。
已知Cry1晶体蛋白家族成员表现出对抗鳞翅目昆虫幼虫的生物活性，并可用作控制鳞翅目昆虫害虫的试剂。Cry1δ-内毒素的前体形式由两个约相同大小的部分组成。前体蛋白的羧基末端部分或毒素原部分稳定晶体形成，且不表现杀虫活性。前体蛋白氨基末端这一半含有Cry1蛋白的毒素部分，且基于Cry1家族成员中保守或基本保守序列的比对，其可进一步再分为三个结构域。这三个亚结构域以Cry1A δ-内毒素的三维晶体学结构模型为基础，其中如从蛋白毒素部分氨基末端所测量的，三个亚结构域分别称作结构域I、结构域II和结构域III。结构域I含有活性毒素部分的大约第一个三分之一，且已显示对于通道形成是关键的(Thompson等人，1995)。结构域II和III各含活性毒素部分的大约中部和羧基末端部分。依赖于所检查的昆虫和δ-内毒素，结构域II和III均与受体结合和昆虫物种特异性牵连(Thompson等人，1995)。
从本领域中已知的众多天然杀虫晶体蛋白结构域结构重配而任意产生性质增强的嵌合蛋白的可能性是渺茫的。这是蛋白质结构、折叠、寡聚化和活化的复杂本质的结果，所述活化包括对嵌合前体(如果以这种形式表达的话)的正确蛋白酶解加工以释放杀虫毒素部分。只有通过各亲本蛋白内针对包括入嵌合结构的具体靶区作出仔细的选择才能构建出与嵌合体所来源的亲本蛋白相比表现有改进的杀虫活性的功能性杀虫毒素。经验已显示，毒素结构域的重装配，即由任意两个或更多个彼此不同的毒素的结构域I、II和III组成的嵌合毒素的装配，导致构建出晶体形成有缺陷和/或完全缺乏针对优选靶昆虫害虫物种的任何可检测的杀虫活性的蛋白。在某些情况下，嵌合毒素将表现出良好的晶体形成特性，但仍不表现可检测的杀虫活性。只有通过尝试与错误才能配制有效的杀虫嵌合体，且尽管那样，本领域技术人员仍不必定最后得到表现与该嵌合体组分所来源的任何单个亲本毒素蛋白相比相同或改进的杀虫活性的嵌合体。
文献报道了从两个或更多个Bt杀虫晶体蛋白前体构建或装配嵌合蛋白的例子，但是并非全部都表现与嵌合体所来源的前体蛋白相比等同或改进的杀虫或晶体形成特性(Bosch等人(WO95/06730)；Thompson等人(WO95/30753)；Thompson等人(WO95/30752)；Malvar等人(WO98/22595)；Gilroy等人(美国专利号5,128,130)；Gilroy(美国专利号5,055,294)；Lee等人(1992)Gene 267：3115-3121；Honee等人(1991)Mol.Microbiol.5：2799-2806；Schnepf等人(1990)J.Biol.Chem.265：20923-20930；Perlak等人(1990)Bio/Technol.8：939-9943；Perlak等人(1993)Plant Mol.Biol.22：313-321)。
已经证明苏云金芽孢杆菌δ内毒素在转基因玉米植物中的表达是控制农业重要昆虫害虫的有效方法(Perlak等人1990；1993)。表达苏云金芽孢杆菌δ内毒素的转基因作物能够使种植者显著减少与施用局部应用的化学杀虫剂有关的时间和成本。尤其有利的是编码苏云金芽孢杆菌δ内毒素的转基因的使用。在严重昆虫压力的地区中表达苏云金芽孢杆菌δ内毒素的作物植物表现出比其它方面类似的非转基因商业植物品种更好的改进产量。然而，据预测，昆虫将进化出针对转基因植物中表达的苏云金芽孢杆菌δ内毒素的抗性。这样的抗性，若广泛流传的话，无疑将限制含有编码这样的苏云金芽孢杆菌δ内毒素基因的种质的商业价值。提高转基因杀虫剂对抗靶害虫的有效性并同时减弱杀虫剂抗性害虫的发展的一种可能途径可以是确保转基因作物表达高水平的苏云金芽孢杆菌δ内毒素(McGaughey和Whalon 1993；Roush1994)。此外，具有有效对抗多组昆虫害虫且通过不同作用模式显示其作用的杀虫基因库可以抵御任何抗性的发展。两种或更多种对相同昆虫物种具有毒性的杀虫组合物在植物中的表达，各杀虫剂以足够高以有效延迟抗性开始的水平表达，可是实现控制抗性发展的另一途径。这样的可用于这样的组合中的杀虫剂的例子包括但不限于Bt毒素、致病杆菌属物种(Xenorhabdus sp.)或光杆状菌属物种(Photorhabdus sp.)杀虫蛋白、脱变态反应和去糖基化patatin蛋白和/或permuteins、植物凝集素等。实现在相同植物中共表达多种杀虫活性蛋白，和/或那些杀虫蛋白的高表达水平而不产生所不期望的植物形态学效果，是难以捉摸的。
从苏云金芽孢杆菌物种中已鉴定的超过二百五十种个别的杀虫蛋白中只有少数已被测试在植物中的表达。几种Cry1’s、Cry3’s、Cry2Aa和Cry2Ab、二元毒素Cry33/34和Cry23/37、以及Cry9已在植物中成功表达。Cry1蛋白代表已在植物中表达但未以高水平表达的最大类蛋白。为避免不期望的植物毒性作用，必须将Cry2Ab靶向叶绿体。大部分种植重组植物的土地表达Cry1A蛋白。所靶向的昆虫害虫物种对Cry1A蛋白的抗性开始的可能性大大高于若抗性管理等位基因也与cry1等位基因一起表达，或者若cry1等位基因以高水平表达的可能性。因此，需要开发出在植物中表达的替代毒素基因作为那些目前在第一和第二代转基因昆虫抗性植物中使用的毒素基因的补充或替代。
发明内容
本发明提供在植物中表达的分离的核苷酸序列，其编码表现鳞翅目昆虫抑制特性的杀虫蛋白。SEQ ID NO：1是这样的核苷酸序列的例子，其由cry1A.105基因组成且编码昆虫抑制性Cry1A.105蛋白。SEQID NO：1与SEQ ID NO：3类似，二者均编码Cry1A.105蛋白。SEQ IDNO：1优选用于双子叶细胞中，而SEQ ID NO：3优选用于单子叶细胞中。SEQ ID NO：4由SEQ ID NO：3编码，且与SEQ ID NO：2的氨基酸序列相同。该分离的核苷酸序列意欲包括表现与SEQ ID NO：1所示序列至少约88％至约90％或更大的核苷酸序列同一性，或在严格杂交条件下与SEQ ID NO：1杂交的序列。该分离的核苷酸序列还意欲包括表现与SEQ ID NO：3所示序列至少约90％核苷酸序列同一性，或在严格杂交条件下与SEQ ID NO：3杂交的序列。
本发明还提供表现针对鳞翅目昆虫物种的抑制活性的分离和纯化的杀虫蛋白。该杀虫蛋白在本文中至少被称为Cry1A.105毒素部分，且表现SEQ ID NO：2所示氨基酸序列。由SEQ ID NO：2中所示的约1177个氨基酸组成的全长前体蛋白也称作杀虫Cry1A.105蛋白，然而任何表现杀虫生物活性的前体蛋白片段也称作杀虫Cry1A.105蛋白，且至少包括对应于SEQ ID NO：2中所示约氨基酸1至约氨基酸612的氨基酸序列部分的Cry1A.105杀虫蛋白，以及还可能包括自约氨基酸2至约氨基酸610的部分。由杀虫有效量的杀虫蛋白组成的任何组合物也意欲包括在本发明范围内。
本发明还提供用于在宿主细胞中表达如SEQ ID NO：2所示的杀虫蛋白的表达盒。该表达盒优选包含在目的宿主细胞中具有功能的启动子，其与编码Cry1A.105蛋白的杀虫部分的核苷酸序列相连并调节其表达。本文中提供如SEQ ID NO：5和SEQ ID NO：7所示的示例性表达盒，其意欲分别用于双子叶植物细胞或单子叶植物细胞。启动子与编码序列可操作性相连并在宿主细胞中共同发挥功能。该表达盒可预定用于任何宿主细胞，但优选用于细菌细胞、真菌细胞、哺乳动物细胞或植物细胞。细菌细胞优选选自芽孢杆菌属(Bacillu)物种细胞、肠杆菌科(Enterobacteriacae)物种细胞、假单胞菌属(Pseudomona)物种细胞、梭菌属(Clostridium)物种细胞、和根瘤菌属(Rhizobium)物种细胞、和土壤杆菌属(Agrobacterium)物种细胞。如果宿主细胞是植物细胞，则优选其是选自作物物种植物细胞的细胞，优选为双子叶植物或单子叶植物细胞。双子叶植物细胞的例子是苜蓿、苹果、杏、芦笋、豆、浆果、黑莓、蓝莓(blueberry)、低芥酸菜子、胡萝卜、花椰菜、芹菜、樱桃、鹰嘴豆、柑橘(citrus tree)、棉花、豇豆、蔓越橘、黄瓜、葫芦、茄子、果树、葡萄、柠檬、莴苣、亚麻子、瓜、芥子、产干果的树、秋葵、橘(orange)、豌豆、桃、花生、梨、李、马铃薯、大豆、南瓜、草莓、甜菜、向日葵、甘薯、烟草、西红柿、芜菁和蔬菜。单子叶植物细胞的例子是玉米、小麦、燕麦、稻、高粱、买罗高粱、荞麦、黑麦、草(羊茅、梯牧草、雀麦草、鸭茅、圣奥古斯丁草(St.Augustine)、狗芽根、剪股颖)和大麦。意欲在植物细胞中使用的表达盒通常包含可操作地连接的序列，所述序列调节比如Cry1A.105杀虫蛋白这样的目的物质的表达水平和效率。这样的序列可能是表达增强子序列、非翻译前导序列、内含子序列、叶绿体靶向肽编码序列和转录终止和多腺苷酸化序列。
该表达盒优选整合进用于稳定Cry1A.105编码序列在宿主细胞中的维持的载体。载体可以是许多本领域已知的结构，但通常是在整合进宿主细胞之前即已在其中构建或插入了表达盒的质粒或复制子。载体意欲包括但不限于质粒、粘粒、杆状病毒穿梭载体、噬粒、YAC、BAC、自杀载体、插入序列、转座子、或甚至与表达盒相连的或其中插入表达盒的线性核苷酸序列。
对鳞翅目昆虫侵袭有抗性的转基因植物是本发明的实施方案。这样的植物包含编码SEQ ID NO：2所示至少约氨基酸2至约氨基酸612的Cry1A.105杀虫蛋白的核苷酸序列。该转基因植物对于控制由比如卷叶蛾(leaf roller)、地老虎(cutworm)、黏虫(armyworm)、钻蛀虫(borer)、避债虫(bagworm)和任何草料进食者(forage feeder)这样的昆虫引起的鳞翅目昆虫侵袭中有效。优选的害虫是草地夜蛾、玉米螟、棉铃叶蛾(corn earworms)(棉铃虫(cotton bollworms))、西南玉米杆草螟和小地老虎。本发明意欲包括由本发明的转基因植物产生的子代和种子或果实或产物，只要编码Cry1A.105杀虫部分的本发明核苷酸序列维持在植物、其子代、种子等的细胞的可遗传和/或质体基因组中。
本发明还提供一种或多种通过在昆虫害虫食物中提供包含杀虫有效量杀虫Cry1A.105蛋白的组合物来控制鳞翅目昆虫侵袭植物的方法。一种这样的组合物可是已源于或源于用编码SEQ ID NO：2所示Cry1A.105氨基酸序列杀虫部分的核酸序列转化的植物细胞的植物细胞。从已被转化以包含表达盒的植物细胞产生的转基因植物将是提供昆虫食物中的杀虫组合物的一种方法，所述表达盒的示例为SEQ IDNO：5和SEQ ID NO：7所示，其包含编码Cry1A.105杀虫氨基酸序列的序列。另一种方法将是在细菌或真菌细胞中产生杀虫有效量的Cry1A.105蛋白并在一种或多种对Cry1A.105蛋白易感的靶昆虫害虫的食物中提供细菌或真菌细胞或纯化量的Cry1A.105蛋白。
提供了鉴定生物样品中编码Cry1A.105氨基酸序列的核苷酸序列的方法。该方法由下述组成，将待测试其中存在Cry1A.105编码序列的样品与特异性与Cry1A.105编码序列结合的多核苷酸探针接触。具体地，该探针序列在严格杂交条件下与Cry1A.105编码序列结合或杂交。对反应混合物中的结合的检测对于Cry1A.105编码序列的存在有诊断价值。
还提供了鉴定样品中Cry1A.105蛋白杀虫片段的方法。该方法由下述组成，将待测试其中存在Cry1A.105杀虫片段的样品与特异性结合杀虫片段的抗体接触。对反应混合物中的结合的检测对于样品中Cry1A.105蛋白的存在有诊断价值。
还提供了嵌合或杂合杀虫蛋白。这样的杂合体由两个或更多个不同的杀虫蛋白组成，各杀虫蛋白表现出针对相同昆虫物种至少一个成员的杀虫活性。杂合杀虫蛋白由各不同杀虫蛋白的部分组成。构建杂合体中所使用的杀虫蛋白部分由至少约50至至少约200个邻接氨基酸组成，所述邻接氨基酸选自组成不同杀虫蛋白之中的任一的邻接氨基酸。SEQ ID NO：2中约第2位氨基酸至约第612位氨基酸所示的Cry1A.105杀虫蛋白意欲包括在，从其中可以选择出用于构建杂合杀虫蛋白的部分的不同杀虫蛋白组内。
本发明的各种优点和特点是显而易见的，通过参考下述详细说明、实施例以及所附的权利要求书，可以更加清楚地理解本发明的本质。
序列简述
SEQ ID NO：1是优选在双子叶植物细胞中表达Cry1A.105杀虫蛋白的合成序列。
SEQ ID NO：2是从SEQ ID NO：1所示核苷酸序列编码得到的Cry1A.105蛋白。
SEQ ID NO：3是优选在单子叶植物细胞中表达Cry1A.105杀虫蛋白的合成序列。
SEQ ID NO：4是从SEQ ID NO：3所示核苷酸序列编码得到的Cry1A.105蛋白。
SEQ ID NO：5代表由在植物细胞中，且优选在双子叶植物细胞中，发挥表达Cry1A.105杀虫蛋白功能的表达盒组成的核苷酸序列。
SEQ ID NO：6代表由SEQ ID NO：5所示的表达盒内的部分编码的Cry1A.105杀虫蛋白。
SEQ ID NO：7代表由在植物细胞中，且优选在单子叶植物细胞中，发挥表达Cry1A.105杀虫蛋白功能的表达盒组成的核苷酸序列。
SEQ ID NO：8代表由SEQ ID NO：7所示的表达盒内的部分编码的Cry1A.105杀虫蛋白。
具体实施方式
根据本发明，本发明人已经构建了编码本文中鉴定为Cry1A.105蛋白的新杀虫蛋白的核苷酸序列。已鉴定，SEQ ID NO：2中所示Cry1A.105氨基酸序列表现以下特性：其提供高于对鳞翅目昆虫物种有毒性的天然存在Bt杀虫蛋白的益处。具体地，Cry1A.105蛋白在单子叶和双子叶植物中均能以高水平表达，而不出现由于与天然存在的Cry1蛋白在植物中表达时所观察到的作用相比表达水平的提高而表现植物毒性作用的大多数转基因事件。此外，Cry1A.105蛋白在苏云金芽孢杆菌中表达时形成稳定的晶体，这可能是由于与嵌合Cry1A.105蛋白的毒素部分相连的Cry1Ac毒素原部分的稳定作用。此外，Cry1A.105杀虫蛋白表现出用迄今为止已鉴定的其它天然存在的Cry1蛋白未观察到的针对鳞翅目物种的一定范围的杀虫生物活性。因此，Cry1A.105蛋白在转基因植物中的表达导致形态学正常的表达更高水平Cry1毒素类似物的转基因事件数目提高，对于被选择用于商业开发的任何事件，所述类似物表现对鳞翅目昆虫害虫物种的广范围控制。这样的事件应当导致产生延迟对Cry1A毒素类似物抗性开始的优势，且当其与第二种毒素组合时，预计发展出对任一毒素的抗性的任何可能性都是非常渺茫的，所述第二种毒素对一种或多种昆虫害虫物种(Cry1A类似物对其也具有毒性)有毒性且以不同于Cry1A类似物的作用模式发挥作用。
本发明已构建了至少两种不同的用于植物的核苷酸序列，各核苷酸序列编码相同的Cry1A.105杀虫蛋白。Cry1A.105蛋白的杀虫部分的第一个(或氨基末端)约三分之二由源自Cry1Ab氨基酸序列的氨基酸序列组成。该序列与毒素部分的羧基末端以及毒素原结构域的部分相连，所述毒素原结构域具有源自从Ecogen Bt aizawai菌株EG6346获得的杀虫Cry1蛋白的氨基酸序列(Chambers等人，1991，J.Bacteriol.173：3966-3976)。Cry1A.105毒素部分然后与基本上为Cry1Ac毒素原肽序列的部分相连。本发明人证实，该结构提供了独特的氨基酸序列，其与该嵌合体所来源的蛋白所表现的特性相比表现出惊人的改进的杀虫特性。此外，Cry1A.105前体蛋白表现出优越的晶体形成特性，而且在具体靶向的鳞翅目昆虫害虫的肠中有效溶解和加工成活性毒素形式。
已经使用美国专利号5,500,365和5,689,052中所述方法构建了本文中所包含的核苷酸序列，具体是通过避免在编码序列中的已经被观察到对于在植物细胞中表达异源基因序列有问题的特定不利序列。编码Cry1A.105蛋白毒素部分的部分或多或少由SEQ ID NO：1和SEQ IDNO：3约第1位至约第1830位所示的核苷酸组成。构建了用于双子叶植物物种，且具体是用于棉花植物的SEQ ID NO：1所示序列。构建了用于在单子叶植物中，且具体是在玉黍蜀或玉米植物物种中表达的示于SEQ ID NO：3的序列。
本发明的核苷酸序列彼此表现约94.3％的总体同一性，且从约第1330位核苷酸至约第3534位核苷酸彼此相同。编码Cry1A.105蛋白毒素部分的这些核苷酸序列的每一个的部分在约第1位核苷酸至约第1830位核苷酸彼此表现约88.9％同一性。编码Cry1A.105蛋白前两个结构域结构的这些核苷酸序列的部分更加多样，且彼此表现仅约84.7％同一性。
本发明人已经用这些序列构建了转基因植物事件。
将SEQ ID NO：1导入含有表达盒的质粒载体，所述表达盒由与碧冬茄(Petunia hybrida)Hsp70非翻译前导序列(Ph.Hsp70，又名DnaK)、鼠耳芥(Arabidopsis thaliana)核酮糖二磷酸羧化酶小亚基叶绿体靶向肽编码序列、和豌豆(Pisum sativum)E9核酮糖二磷酸羧化酶小亚基基因转录终止和多腺苷酸化序列可操作地连接的增强的玄参花叶病毒(Figwort Mosaic Virus)启动子(eFMV)序列组成。将SEQ ID NO：1所示Cry1A.105编码序列以与靶向肽编码序列3’末端编码序列符合读框地插入该表达盒中，并使其与靶向肽编码序列3’末端编码序列直接接近且位于E9终止序列上游。所得表达盒的核苷酸序列示于SEQ IDNO：5。切除含有与包含植物可表达GUS标记的第二个表达盒连接的Cry1A.105表达盒的载体部分，并用生物射弹方法将其用于产生转基因棉花事件。在生物测定中测试转基因事件对抗几种不同鳞翅目害虫物种的杀虫活性，且确定转基因事件比以前存在的仅包含Cry1Ac或Cry1Ac和Cry2Ab蛋白的组合的昆虫抗性棉花植物表现显著更好的昆虫控制特性。此外，一些Cry1A.105转基因棉花事件在整个种植季，甚至在棉铃中，表现的Cry1A.105蛋白累积水平超过10～20ppm，且对植物或生殖组织不表现任何植物毒性作用。这与从前已经测试过的其它Cry1蛋白形成对照，无论是否靶向叶绿体，所述其它Cry1蛋白通常仅能达到低于约10ppm的累积水平。在棉花中测试其它Cry1类型蛋白时也观察了植物毒性作用，尤其是在Cry1累积水平达到或超过约10ppm的时候。
将SEQ ID NO：3导入含有表达盒的质粒载体，所述表达盒由与普通小麦(Triticum aestivum)主要叶绿素a/b结合蛋白基因非翻译前导序列和稻(Oryza sativa)肌动蛋白内含子序列，和普通小麦hsp 17基因转录终止和多腺苷酸化序列可操作地连接的增强的花椰菜花叶病毒启动子(eCaMV)序列组成。将示于SEQ ID NO：3的Cry1A.105编码序列插入该表达盒中，使其与内含子序列3’直接接近且位于终止序列上游。所得表达盒的核苷酸序列示于SEQ ID NO：7。该载体还包含用于选择用Cry1A.105表达盒转化的事件的草甘膦除草剂选择标记。在生物测定中针对几种鳞翅目害虫物种测试用Cry1A.105表达盒转化后选择的玉黍蜀事件，并确定它们表现用比如Cry1Ab这样的其它Bt杀虫蛋白转化的事件中不普遍的广范围杀虫活性。表达杀虫水平的Cry1A.105的事件所表现的草地夜蛾和小地老虎的活性，与表达Cry1Ab的事件针对棉铃叶蛾和玉米螟的杀虫活性相比与之相当或更强的Cry1A.105杀虫活性偶联，为Cry1A.105事件提供了更广谱的杀虫活性。
本发明的核苷酸序列是示例性的。其它的核苷酸序列能够在植物细胞中表达Cry1A.105杀虫蛋白片段，而且其它在其它类型的宿主细胞中很好表达的核苷酸序列也能够被设计。不限制本发明公开内容的范围，用于表达Cry1A.105杀虫片段的核苷酸序列意欲与本文所例举的核苷酸序列表现至少约85％、或至少约90％、或至少约95％、或至少约99％或更大的核苷酸序列同一性。意欲用于在除植物细胞之外的宿主细胞中表达Cry1A.105杀虫片段的其它核苷酸序列可以与例举的核苷酸序列具有任何百分比的同一性或相似性。由于遗传密码的冗余，核苷酸序列可以发生变化，且由此有可能合成任何数目的编码SEQ IDNO：2所示氨基酸序列任意部分的核酸序列，且所有这些序列都意欲在本发明范围内。任何至少编码Cry1.105蛋白杀虫片段的分离和纯化的核酸序列，以及可以通过抗体、核酸探针或一对或多对被设计用于产生由这样的序列组成的扩增子的引物来检测其中的核酸的任何组合物，都意欲包括在本公开内容范围内。
本文例举的且在玉黍蜀中表达的核酸序列仅由Cry1A.105前体蛋白编码序列组成，而在棉花中表达的序列由叶绿体靶向的Cry1A.105前体蛋白编码序列组成。已经证明Cry1蛋白在植物中的表达是有问题的。不知任何特定的Cry1蛋白是否将在任何特定的植物中良好表达，因此需要尝试与错误试验。一些在玉米中表达的Cry1蛋白将导致产生植物毒性作用，且因此将该蛋白靶向叶绿体有时减轻这样的作用。在表达Cry1蛋白的棉花植物中也观察到类似的情况。本文的例子无意于教导如果定位于胞质空间，则Cry1A.105表达只可能在玉米中，以及类似地，无意于教导如果定位于质体，Cry1A.105表达只可能在棉花中。这些实施例意欲教导，任一蛋白质定位方法都与该蛋白一起发挥功能以实现形态学正常的植物，所述植物表现高水平Cry1A.105蛋白表达和累积，以及对选自叶蛾属(Anticarsia)、尺叶蛾属(Pseudoplusia)、Rachiplusia、Helicoverpa、实叶蛾属(Heliothis)、灰翅叶蛾属(Spodoptera)、叶小卷娥属(Epinotia)和Armigera这些属的广范围鳞翅目昆虫植物害虫表现商业水平的抗性。据信，任意质体靶向肽编码序列将有效地发挥将前体Cry1A.105蛋白导向质体/叶绿体的作用。
非翻译前导序列、内含子和3’转录终止以及多腺苷酸化序列是本领域已知的，且本领域技术人员能理解，在特定的情况下，通过将这些序列整合进表达盒中可增强或者稳定表达。许多这样的序列是本领域已知的，且意欲将它们包括在本发明的公开内容范围内。类似地，对所连接的序列发挥功能以实现调节的表达的启动子是本领域已知的，且也意欲包括在本发明的公开内容范围内。可选择启动子用于在许多参数组合中驱动所连接序列的表达，包括但不限于表达的时间控制、表达的空间或组织特异性控制、以及控制需要在特定植物细胞或组织中累积的特定基因产物的量。
含有Cry1A.105氨基酸序列杀虫片段的分离和纯化的蛋白也意欲包括在本发明范围内。变体也意欲包括在本发明范围内，只要影响该变异的一种或多种氨基酸取代对于被取代的氨基酸总体上是保守的，且该(这些)取代不导致杀虫生物活性或物种特异性范围的降低。Cry1A.105蛋白杀虫片段欲指，SEQ ID NO：2约第1位氨基酸至约第650位氨基酸、或约第2位氨基酸至约第612位氨基酸、或约第5位氨基酸至约第610位氨基酸、或约第10位氨基酸至约第600位氨基酸所示氨基酸序列的部分。作为选择，Cry1A.105蛋白的杀虫片段欲由约550至约650个邻接氨基酸组成，所述邻接氨基酸选自SEQ ID NO：2所示第1位至约第650位氨基酸残基。由第1位至约第3534位残基组成的全长前体蛋白，表现优良的晶体形成特性，并由双子叶和单子叶植物物种良好耐受。该前体蛋白在结晶形式下还表现出良好的稳定性，以及在碱性pH下还表现良好的溶解度，具体是在约8.0至约12.0、或约8.5至约11.5、或约pH 9.0至约pH 11.0的碱性pH下。
本发明的蛋白可纯化并以杀虫有效量单独用于许多意欲用作鳞翅目害虫控制试剂的组合物中，或可以以杀虫有效量与许多不同于Cry1A.105蛋白的其它杀虫剂组合使用。这样的其它的杀虫剂意欲包括但不限于其它Bt Cry或其它无论对于鳞翅目物种是否具有毒性的杀虫组合物，包括化学杀虫剂、杀真菌剂或抑真菌剂、抗生素、抗细菌剂、制菌剂、和杀线虫剂或制线虫剂(nematostatic agent)。这样的包括Cry1A.105以及许多其它杀虫剂的杀虫剂组合可通过转基因细胞生产，或用纯化或基本纯化的杀虫剂配制成如下形式的杀虫剂组合物，所述形式由灰尘、颗粒材料、油悬浮液、水悬浮液、油和水乳状液的混合物、或可湿性粉末组成，然后再于用于叶施用的农业可接受载体中提供。也可以将这些组合物配制成种子处理，抑或与欲包含在种子处理中的组合物中的Cry1A.105一起，抑或作为应用于来自已被转化以表达杀虫有效量Cry1A.105的转基因植物的种子的组合物，从而将包含杀虫剂的种子处理组合物连同从所述种子生长的植物的细胞一起提供给靶鳞翅目害虫，所述细胞产生杀虫有效量的Cry1A.105蛋白。各自对相同的昆虫物种有毒性但通过不同的作用模式表现其毒性作用的杀虫蛋白组合对于控制鳞翅目物种或者延迟对任一单种杀虫剂的抗性开始是尤其有用的杀虫剂组合，在未开始抗性前所述任一单种杀虫剂有效对抗特定的鳞翅目物种。这样的蛋白的示例性组合是，本发明的Cry1A.105蛋白，即第一杀虫蛋白，与至少第二种不同于第一蛋白的杀虫蛋白偶联。这样的不同杀虫蛋白包括但不限于其它鳞翅目Bt.结晶蛋白(其它Cry1’s、Cry2’s、Cry5’s、Cry9’s)、VIP蛋白、称作TIC蛋白的鳞翅目杀虫蛋白以及由细菌的致病杆菌属和光杆状菌属物种产生的杀虫蛋白。向昆虫害虫的食物中提供一种或多种杀虫蛋白以及设计用于实现dsDNA介导的对昆虫存活所必需的一种或多种基因的基因抑制的试剂的组合，对于控制鳞翅目物种或者延迟对任一单种杀虫剂的抗性的开始是尤其有用的杀虫剂组合，在未开始抗性前所述任一单种杀虫剂有效对抗特定的鳞翅目物种。
用本发明的核苷酸序列转化的植物也是本发明提供的另一实施方案。将DNA稳定导入植物细胞的方法是本领域已知的，且包括但不限于真空过滤、土壤杆菌或根瘤菌介导的转化、电穿孔和各种冲击法。导入植物中的DNA通常靶向插入核染色体中，尽管也可以实现插入叶绿体或质体DNA中。导入植物中的DNA通常与为鉴定或选择已被目的DNA稳定转化的一种或多种细胞提供工具的序列连接或结合，所述序列包括但不限于可计分标记，比如荧光或光发射基因和编码在适当底物存在下赋予被转化的一种或多种细胞以比色特征的色素或酶的基因，或者通过包括允许对被转化细胞和组织进行阳性选择的选择标记，从而为被转化细胞提供生长优势并基本上使未转化细胞或组织变为静止或死亡。这样的选择标记包括但不限于编码草铵膦、bar、氨甲蝶呤抗性、新霉素磷酸转移酶、草甘膦不敏感的EPSPS酶、草甘膦氧化还原酶(GOX)酶、大肠杆菌(E.coli)phnO或其等价物等的基因。
本发明范围内还包括载体和其它类型的序列，它们被设计用于在实验室中操纵时维持、操纵和/或引导示例核苷酸序列，或者设计用于导入宿主细胞中，这些载体和其它类型的序列意欲包括但不限于噬菌体、质粒、杆状病毒穿梭载体、yacmids、粘粒等。
转化的植物也在本发明范围内。本发明公开内容尤其使被转化以含有编码至少Cry1A.105蛋白杀虫片段的核苷酸序列的植物成为可能。单子叶和双子叶植物都预计在本发明的范围内。单子叶植物意欲包括但不限于玉米、小麦、燕麦、稻、高粱、买罗高粱、荞麦、黑麦、草(羊茅、梯牧草、雀麦草、鸭茅、圣奥古斯丁草、狗芽根、剪股颖)和大麦，且双子叶植物意欲包括至少苜蓿、苹果、杏、芦笋、豆、浆果、黑莓、蓝莓、低芥酸菜子、胡萝卜、花椰菜、芹菜、樱桃、鹰嘴豆、柑橘、棉花、豇豆、蔓越橘、黄瓜、葫芦、茄子、果树、葡萄、柠檬、莴苣、亚麻子、瓜、芥子、产干果的树、秋葵、橘、豌豆、桃、花生、梨、李、马铃薯、大豆、南瓜、草莓、甜菜、向日葵、甘薯、烟草、西红柿、芜菁和蔬菜。这些植物的产物以及由这些植物产生的种子和组织明确包括在本发明内，只要该种子、组织或产物包含编码Cry1A.105蛋白杀虫片段的转基因。
本发明提供了在生物样品中检测Cry1A.105蛋白或编码Cry1A.105蛋白杀虫片段的核苷酸序列的方法。Cry1A.105可用于免疫动物以产生特异于Cry1A.105表位的抗体。Cry1A.105特异性抗体可用于检测生物样品中Cry1A.105的存在。检测抗体与抗原结合的方法是本领域已知的。检测生物样品中抗体与Cry1A.105表位的结合对样品中蛋白的存在有诊断价值。
也可以检测编码Cry1A.105杀虫片段的核苷酸序列。可以使用合成的核苷酸探针与靶序列，即编码Cry1A.105杀虫片段的核苷酸序列结合。检测探针与靶序列结合的方法是本领域已知的。检测探针与靶Cry1A.105编码序列的结合对样品中编码序列的存在有诊断价值。
合成的核苷酸引物可用于热扩增反应以从被怀疑包含编码Cry1A.105蛋白杀虫片段的核苷酸序列的生物样品中产生扩增子。在这样的热扩增反应中产生的扩增子的存在对样品中该核苷酸序列的存在有诊断价值。作为对检测生物样品中本发明Cry1A.105编码序列的存在有诊断价值的探针特别有用的序列是对应于或完全互补于(1)SEQID NO：1或SEQ ID NO：3所示第1401-1420位的核苷酸，或(2)SEQ IDNO：1或SEQ ID NO：3所示第1821-1840位核苷酸的序列。这些序列对应于(1)跨越编码介于不同杀虫蛋白部分的结构域II和结构域III之间的接头的序列的20个核苷酸，所述结构域用于构建本发明蛋白的杀虫部分，和(2)跨越编码介于不同蛋白编码部分的结构域III和毒素原编码部分之间的接头的序列的20个核苷酸，所述结构域和部分用于构建前毒素原Cry1Ab.105蛋白的编码序列。作为这些DNA部分(1401-1420或1821-1840)任一个的核苷酸序列，或与之互补的核苷酸序列可用作检测生物样品中这些编码序列的存在的探针。对这样的结合的检测对生物样品中这样的编码序列的存在有诊断价值。如本领域技术人员所将认识到的这些DNA部分每一侧侧接的其它序列可用作从这样的生物样品中扩增各种大小的扩增子部分的引物，而且这样的扩增子对样品中这样的编码序列的存在有诊断价值。例如，对应于SEQID NO：1中第1201-1220位所示核苷酸序列的第一引物序列可用作其中第二引物序列对应于SEQ ID NO：1中第1581-1600位所示核苷酸序列的反向互补体的热扩增反应中的正向引物。当这样的引物共同用于对包含SEQ ID NO：1的生物样品的热扩增反应中时，其将导致合成对应于SEQ ID NO：1第1201至第1600位核苷酸的扩增子，即400个核苷酸的扩增子，所述扩增子会包含SEQ ID NO：1所示第1401-1420位的20个核苷酸的部分，且由此将对这样的样品中Cry1A.105编码序列的存在有诊断价值。
提供了在样品中检测Cry1A.105存在或检测编码Cry1A.105的核苷酸序列的存在的试剂盒。连同该试剂盒还提供与实施检测目的试剂的方法所需的所有试剂和对照样品，以及使用说明书。
以下实施例描述本发明的优选实施方案。对于本领域技术人员来说，考虑本文所公开的本发明说明书或实践后，权利要求范围内的其它实施方案也是显而易见的。说明书以及实施例，仅欲被认为是示例性的，且实施例之后所附权利要求书指明了本发明的范围和精神。
实施例
实施例1.
该实施例阐述编码杀虫Cry1A.105蛋白的合成的核苷酸序列。
构建用于双子叶植物中的如SEQ ID NO：1所示的编码Cry1A.105杀虫蛋白的核苷酸序列。氨基酸序列翻译示于SEQ ID NO：2。毒素编码部分或多或少由约第1位至约第1830位核苷酸组成。
构建用于在单子叶植物中表达的如SEQ ID NO：3所示的编码Cry1A.105氨基酸序列的核苷酸序列。氨基酸序列翻译示于SEQ IDNO：4。毒素编码部分或多或少由约第1位至约第1830位核苷酸组成。
SEQ ID NO：1和SEQ ID NO：3所示核苷酸序列彼此基本等同。SEQ ID NO：1和SEQ ID NO：3表现约94.3％的总体同一性。这两条编码序列从约第1330位核苷酸至第3534位核苷酸等同。各序列的毒素编码部分由约第1位的核苷酸至第1830位的核苷酸组成，且这些部分彼此表现约88.9％的同一性。两条序列间的基本差别在于约第1位核苷酸至约第1329位核苷酸，或编码Cry1A.105蛋白毒素部分的约第一个三分之二的部分。这两条序列贯穿该部分表现约84.7％的同一性。
根据《国际承认用于专利程序的微生物保存布达佩斯条约》，于2005年8月31日将用名称为pMON70522的质粒转化的大肠杆菌菌株(TOP10，Invitrogen，Inc.)保藏于位于1815North University Street，in Peoria，Illinois 61604 U.S.A.的国际保藏单位农业研究机构保藏中心(Agriculture Research Culture Collection)(NRRL)，并命名为NRRLB-30873，所述名称为pMON70522的质粒包含β-内酰胺酶选择标记和SEQ ID NO：3所示编码Cry1A.105的序列。
实施例2
该实施例阐述表达Cry1A.105蛋白的转基因棉花植物。
将Delta和Pineland DP50棉花种子表面灭菌，并过夜萌发。分离分生组织外植体，且通过显微解剖除去初生叶。将解剖的外植体放入靶向培养基(targeting medium)中，从而使分生组织定向为与粒子递送(particle deliyery)方向垂直。转化载体，pMON47740，含有带有SEQ ID NO：9所示核苷酸序列的表达盒。从该质粒中切除KpnI片段，用HPLC分离，并用于枪(gun)转化棉花分生组织外植体，所述KpnI片段包含在e35S启动子控制下的GUS标记基因和在eFMV启动子控制下的叶绿体靶向的Cry1A.105编码序列。将包含Cry1A.105表达盒和GUS标记两者的纯化的DNA沉淀在显微金珠上，并在Mylar片(sheet)上包被成薄层。在部分真空下，通过放电式粒子递送(electricdischarge particle delivery)将DNA加速进分生组织。轰击后，将外植体去靶向(de-target)到不含选择剂的无激素培养基中。对来自再生小植物的叶组织取样，并测定GUS标记的表达。将表现高水平GUS表达的转基因植物送到温室中作进一步筛选。再次测试这些植物中GUS的表达，并剪除植物中阴性的部分。重复进行取样和剪除GUS-阴性组织的循环，直到取自各植物的所有部分都是GUS标记阳性的。之后在标准温室条件下维持这些植物直到收获种子。
用生物测定测试从F1GUS阳性转基因棉花植物获得的组织针对棉铃虫(CBW)和草地夜蛾(FAW)的杀虫活性。将以前产生的表达杀虫水平的Cry1Ac或Cry1Ac与Cry2Ab的组合的同基因棉花植物用作阳性对照，且将非转基因同系(isoline)用作阴性对照。
将CBW棉蕾测定(square assays)用作确定转基因棉花植物杀虫活性的一种方法(Adamczyck等人，(2001)J.Econ.Entomol.94：284-290；Kranthi等人(2005)Current Science 89：291-298)。收集叶组织棉蕾(火柴头大小或更大)并单独置于测定孔中。各棉蕾用一个三龄CBW幼虫侵袭。侵袭后五天记录存活昆虫的数目。
还采用CBW棉铃测定(boll assays)确定了从转基因植物收集的棉铃组织的杀虫活性。从各事件收集了8个硬绿色棉铃(开花后)并置于单独的杯中，并用三龄CBW幼虫侵袭。侵袭后五天记录存活昆虫的数目。
进行叶测定来确定转基因叶组织对FAW的杀虫活性。从棉花植物末端取新叶。收集两个直径各为约3/4″的叶孔(punch)并将其置于16个单独的测定孔的各孔中。各孔用单个二龄或三龄FAW幼虫侵袭。侵袭后五天记录存活昆虫的数目。
生物测定结果示于表1。结果显示，表达Cry1A.105的转基因棉花事件比表达Cry1Ac或Cry1Ac与Cry2Ab的组合的转基因事件对FAW和CBW均表现更强的杀虫活性。
表1.用转基因棉花植物组织对FAW和CBW的生物测定结果。
还在类似的生物测定中测试了烟夜蛾和棉铃叶蛾。在各情况中，Cry1A.105植物也表现针对这些害虫的杀虫活性。
实施例3
本实施例阐述表达Cry1A.105蛋白的转基因玉米植物。
从用载体pMON40232转化的细胞再生转基因玉米植物。pMON40232包含具有SEQ ID NO：7所示核苷酸序列的表达盒，所述表达盒包含可操作地连接的增强的CAMV 35S启动子、小麦CAB前导序列、稻肌动蛋白1内含子、Cry1A.105编码序列和小麦hsp17基因3’转录终止和多腺苷酸化序列。编码鼠耳芥(Arabidopsis thaliana)EPSPS叶绿体靶向序列(At.EPSES-CTP2)的核苷酸序列位于Cry1A.105编码序列上游且与其符合读框。pMON40232包含编码对除草剂草甘膦不敏感的EPSPS的重组基因，以用于选择转基因事件。将从用pMON40232转化的组织得到的转基因事件命名为LAJ105。筛选转基因事件中任何载体主链的不存在、单个简单插入序列的存在、以及包含编码Cry1A.105蛋白的核苷酸序列的表达盒的完整性。
对满足事件筛选限制的事件作生物测定。在该生物测定中，将LAJ 105转基因玉米植物与同基因LH198阴性对照和表达Cry1Ab蛋白杀虫部分的阳性对照MON810品种作比较。从10个单独的Cry1A.105转基因事件以及对照的每一个获取五个各直径约1厘米的叶盘。将叶盘置于填充有琼脂的孔中以保持植物材料肿胀。之后将叶盘饲喂给FAW、小地老虎(BCW)、玉米螟(ECB)、棉铃叶蛾(CEW)和西南玉米杆草螟(SWCB)新生幼虫。各孔应用一只新生FAW幼虫、一只CEW幼虫、两只新生BCW、两只新生SWCB幼虫或四只新生ECB幼虫。四天后用叶损害率(LDR)等级0-11来评估饲喂损害，0表示没有可见的饲喂损害，11表示至少50％的叶盘被吃掉，且该等级上0至11的各点表示在观察中所观察到的叶盘饲喂损害的5％增长。
生物测定结果表示，表达Cry1A.105蛋白的事件对FAW、ECB和CEW表现的杀虫活性比Cry1Ab对照对相同害虫幼虫所表现的LDR更强。这三种害虫在Cry1A.105事件上的LDR小于1，而Cry1Ab对照表现的LDR范围在约8至约10。对于Cry1A.105事件和Cry1Ab对照，测试针对SWCB的活性时，其LDR始终为1至2，这表示Cry1A.105蛋白对于SWCB不比Cry1Ab具备更高的毒性。该生物测定的结果支持了表明Cry1Ab对于控制BCW无效这一从前的结果。Cry1A.105事件对抗BCW并不比Cry1Ab对照更有效。因此，以Cry1A.105蛋白在植物中(in planta)的表达水平，这些植物在控制包括但不限于叶蛾属、尺叶蛾属、Rachiplusia、实叶蛾属、Helicoverpa、灰翅叶蛾属、叶小卷娥属和Armigera属中的其它鳞翅目属植物害虫中是有效的。
Claims (16)
1.控制转基因植物中灰翅叶蛾属昆虫侵袭并提供昆虫抗性控制的方法，其包括在植物中表达至少两种不同的对灰翅叶蛾属物种/昆虫有毒性的杀虫蛋白。
2.权利要求1的方法，其中所述至少两种不同的杀虫蛋白包括杀灰翅叶蛾属昆虫的VIP蛋白和杀灰翅叶蛾属昆虫的Cry1蛋白。
3.权利要求2的方法，其中作为所述表达的结果，通过延迟摄食所述植物的灰翅叶蛾属昆虫群体中对所述VIP和Cry1蛋白的昆虫抗性的开始而提供所述昆虫抗性控制。
4.权利要求1的方法，其中所述转基因植物是选自玉米、小麦、燕麦、稻、高粱、买罗高粱、荞麦、黑麦、羊茅、梯牧草、雀麦草、鸭茅、圣奥古斯丁草、狗芽根、剪股颖和大麦的单子叶植物。
5.权利要求1的方法，其中所述转基因植物是选自苜蓿、苹果、杏、芦笋、豆、浆果、黑莓、蓝莓、低芥酸菜子、胡萝卜、花椰菜、芹菜、樱桃、鹰嘴豆、柑橘、棉花、豇豆、蔓越橘、黄瓜、葫芦、茄子、果树、葡萄、柠檬、莴苣、亚麻子、瓜、芥子、产干果的树、秋葵、橘、豌豆、桃、花生、梨、李、马铃薯、大豆、南瓜、草莓、甜菜、向日葵、甘薯、烟草、西红柿、芜菁和蔬菜的双子叶植物。
6.权利要求2的方法，其中所述Cry1蛋白是Cry1A蛋白。
7.权利要求6的方法，其中所述Cry1A蛋白是Cry1A.105蛋白。
8.控制转基因植物中灰翅叶蛾属侵袭同时抵御对所述植物的灰翅叶蛾属昆虫抗性的发展的方法，其包括在所述植物中表达a)杀灰翅叶蛾属的VIP蛋白和b)杀灰翅叶蛾属的Cry1蛋白的组合。
9.充分延迟灰翅叶蛾属群体中对表达杀虫蛋白的转基因植物的昆虫抗性的开始以控制所述昆虫的方法，其包括在所述植物中表达杀所述昆虫的VIP蛋白以及杀所述昆虫的Cry1蛋白。
10.降低对表达杀虫蛋白的转基因植物的灰翅叶蛾属昆虫抗性的出现的可能性以控制所述昆虫物种的方法，其包括在所述植物中表达杀所述昆虫物种的VIP蛋白以及杀所述昆虫物种的Cry1蛋白。
11.播种、种植或生长受到抗草地夜蛾保护的植物的方法，其包括如下步骤：播种、种植或生长包含编码杀灰翅叶蛾属昆虫的VIP蛋白的基因和编码杀灰翅叶蛾属昆虫的Cry1蛋白的基因的植物。
12.权利要求8-11中任一项的方法，其中所述Cry1蛋白是Cry1A蛋白。
13.权利要求12的方法，其中所述Cry1A蛋白是Cry1A.105蛋白。
14.用于转基因植物的有效的灰翅叶蛾属昆虫抗性控制的方法，其包括在所述植物中以高水平共表达两种或更多种对灰翅叶蛾属昆虫有毒性但各自表现出不同的实现其杀灭活性的模式的杀虫蛋白，其中所述两种或更多种杀虫蛋白包括VIP蛋白和Cry1蛋白。
15.权利要求14的方法，其中所述Cry1蛋白是Cry1A蛋白。
16.权利要求15的方法，其中所述Cry1A蛋白是Cry1A.105蛋白。
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104621171A (zh) * | 2015-01-30 | 2015-05-20 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| CN110177464A (zh) * | 2016-12-20 | 2019-08-27 | 孟山都技术公司 | 新型昆虫抑制性蛋白 |
| CN110214189A (zh) * | 2016-09-06 | 2019-09-06 | 农业生物群落股份有限公司 | 杀虫基因和使用方法 |
| CN110622998A (zh) * | 2019-10-14 | 2019-12-31 | 中国农业科学院植物保护研究所 | 一种蛋白在防治草地贪夜蛾和/或斜纹夜蛾中的应用 |
| CN112175093A (zh) * | 2014-10-16 | 2021-01-05 | 孟山都技术有限公司 | 对鳞翅目害虫具有毒性或抑制性的新型嵌合杀昆虫蛋白质 |
Families Citing this family (375)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EG26529A (en) * | 2001-06-11 | 2014-01-27 | مونسانتو تكنولوجى ل ل سى | Prefixes for detection of DNA molecule in cotton plant MON15985 which gives resistance to damage caused by insect of squamous lepidoptera |
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| EP1969934A1 (de) * | 2007-03-12 | 2008-09-17 | Bayer CropScience AG | 4-Cycloalkyl-oder 4-arylsubstituierte Phenoxyphenylamidine und deren Verwendung als Fungizide |
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| EP1969930A1 (de) | 2007-03-12 | 2008-09-17 | Bayer CropScience AG | Phenoxyphenylamidine und deren Verwendung als Fungizide |
| EP1969929A1 (de) | 2007-03-12 | 2008-09-17 | Bayer CropScience AG | Substituierte Phenylamidine und deren Verwendung als Fungizide |
| WO2008110279A1 (de) | 2007-03-12 | 2008-09-18 | Bayer Cropscience Ag | Dihalogenphenoxyphenylamidine und deren verwendung als fungizide |
| JP2010524869A (ja) * | 2007-04-19 | 2010-07-22 | バイエル・クロツプサイエンス・アクチエンゲゼルシヤフト | チアジアゾリルオキシフェニルアミジンおよび殺菌剤としてのこれらの使用 |
| DE102007045957A1 (de) | 2007-09-26 | 2009-04-09 | Bayer Cropscience Ag | Wirkstoffkombinationen mit insektiziden und akarziden Eigenschaften |
| DE102007045956A1 (de) | 2007-09-26 | 2009-04-09 | Bayer Cropscience Ag | Wirkstoffkombination mit insektiziden und akariziden Eigenschaften |
| DE102007045920B4 (de) | 2007-09-26 | 2018-07-05 | Bayer Intellectual Property Gmbh | Synergistische Wirkstoffkombinationen |
| DE102007045919B4 (de) | 2007-09-26 | 2018-07-05 | Bayer Intellectual Property Gmbh | Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften |
| DE102007045953B4 (de) | 2007-09-26 | 2018-07-05 | Bayer Intellectual Property Gmbh | Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften |
| DE102007045922A1 (de) | 2007-09-26 | 2009-04-02 | Bayer Cropscience Ag | Wirkstoffkombinationen mit insektiziden und akariziden Eigenschaften |
| EP2090168A1 (de) | 2008-02-12 | 2009-08-19 | Bayer CropScience AG | Methode zur Verbesserung des Pflanzenwachstums |
| EP2072506A1 (de) | 2007-12-21 | 2009-06-24 | Bayer CropScience AG | Thiazolyloxyphenylamidine oder Thiadiazolyloxyphenylamidine und deren Verwendung als Fungizide |
| EP2092824A1 (de) | 2008-02-25 | 2009-08-26 | Bayer CropScience AG | Heterocyclyl-Pyrimidine |
| EP2103615A1 (de) | 2008-03-19 | 2009-09-23 | Bayer CropScience AG | 4'4'-Dioxaspiro-spirocyclisch substituierte Tetramate |
| EA201001699A1 (ru) | 2008-04-30 | 2011-04-29 | Байер Кропсайенс Аг | Сложные эфиры и тиоэфиры тиазол-4-карбоновой кислоты в качестве средств защиты растений |
| EP2307390B1 (de) | 2008-06-27 | 2014-01-22 | Bayer CropScience AG | Thiadiazolyloxyphenylamidine und deren verwendung als fungizide |
| EP2168434A1 (de) | 2008-08-02 | 2010-03-31 | Bayer CropScience AG | Verwendung von Azolen zur Steigerung der Resistenz von Pflanzen oder Pflanzenteilen gegenüber abiotischem Stress |
| MX2011001427A (es) | 2008-08-08 | 2011-06-20 | Bayer Bioscience Nv | Metodos para la caracterizacion e identificacion de fibras vegetales. |
| JP2011530548A (ja) | 2008-08-14 | 2011-12-22 | バイエル・クロップサイエンス・アーゲー | 殺虫性４−フェニル−１ｈ−ピラゾール類 |
| DE102008041695A1 (de) | 2008-08-29 | 2010-03-04 | Bayer Cropscience Ag | Methoden zur Verbesserung des Pflanzenwachstums |
| EP2161259A1 (de) | 2008-09-03 | 2010-03-10 | Bayer CropScience AG | 4-Halogenalkylsubstituierte Diaminopyrimidine als Fungizide |
| KR20110067052A (ko) * | 2008-10-01 | 2011-06-20 | 바이엘 크롭사이언스 아게 | 식물 보호제로서의 헤테로사이클릴 치환된 티아졸 |
| WO2010037482A2 (de) * | 2008-10-02 | 2010-04-08 | Bayer Cropscience Aktiengesellschaft | Verwendung von schwefelhaltigen, heteroaromatischen säureanaloga |
| ES2445540T3 (es) | 2008-10-15 | 2014-03-03 | Bayer Cropscience Ag | Uso de ditiina-tetracarboximidas para combatir hongos fitopatógenos |
| EP2184273A1 (de) | 2008-11-05 | 2010-05-12 | Bayer CropScience AG | Halogen-substituierte Verbindungen als Pestizide |
| EP2201838A1 (de) | 2008-12-05 | 2010-06-30 | Bayer CropScience AG | Wirkstoff-Nützlings-Kombinationen mit insektiziden und akariziden Eigenschaften |
| JP2012511525A (ja) | 2008-12-11 | 2012-05-24 | バイエル・クロップサイエンス・アーゲー | 植物保護剤としてのチアゾリルオキシムエーテルおよびヒドラゾン |
| WO2010069495A1 (de) * | 2008-12-18 | 2010-06-24 | Bayer Cropscience Aktiengesellschaft | Atpenine |
| EP2198710A1 (de) | 2008-12-19 | 2010-06-23 | Bayer CropScience AG | Verwendung von 5-Pyridin-4yl-(1,3)Thiazole zur Bekämpfung phytopathogener Pilze |
| EP2198709A1 (de) | 2008-12-19 | 2010-06-23 | Bayer CropScience AG | Verfahren zur Bekämpfung resistenter tierischer Schädlinge |
| EP2223602A1 (de) | 2009-02-23 | 2010-09-01 | Bayer CropScience AG | Verfahren zur verbesserten Nutzung des Produktionspotentials genetisch modifizierter Pflanzen |
| AU2009335333B2 (en) | 2008-12-29 | 2015-04-09 | Bayer Intellectual Property Gmbh | Method for improved use of the production potential of genetically modified plants |
| EP2204094A1 (en) | 2008-12-29 | 2010-07-07 | Bayer CropScience AG | Method for improved utilization of the production potential of transgenic plants Introduction |
| EP2039770A2 (en) | 2009-01-06 | 2009-03-25 | Bayer CropScience AG | Method for improved utilization of the production potential of transgenic plants |
| EP2039771A2 (en) | 2009-01-06 | 2009-03-25 | Bayer CropScience AG | Method for improved utilization of the production potential of transgenic plants |
| EP2039772A2 (en) | 2009-01-06 | 2009-03-25 | Bayer CropScience AG | Method for improved utilization of the production potential of transgenic plants introduction |
| EP2387317A2 (de) | 2009-01-15 | 2011-11-23 | Bayer CropScience AG | Fungizide Wirkstoffkombinationen enthaltend Bixafen oder dessen Salze |
| WO2010081646A2 (de) | 2009-01-15 | 2010-07-22 | Bayer Cropscience Aktiengesellschaft | Fungizide wirkstoffkombinationen |
| KR20110106448A (ko) | 2009-01-19 | 2011-09-28 | 바이엘 크롭사이언스 아게 | 사이클릭 디온 및 살충제, 살비제 및/또는 살진균제로서의 그의 용도 |
| EP2227951A1 (de) | 2009-01-23 | 2010-09-15 | Bayer CropScience AG | Verwendung von Enaminocarbonylverbindungen zur Bekämpfung von durch Insekten übertragenen Viren |
| JP5592398B2 (ja) | 2009-01-28 | 2014-09-17 | バイエル・クロップサイエンス・アーゲー | 殺菌剤ｎ−シクロアルキル−ｎ−二環式メチレン−カルボキサミド誘導体 |
| AR075126A1 (es) | 2009-01-29 | 2011-03-09 | Bayer Cropscience Ag | Metodo para el mejor uso del potencial de produccion de plantas transgenicas |
| EP2223917A1 (de) | 2009-02-02 | 2010-09-01 | Bayer CropScience AG | Isothiazolyloxyphenylamidine und deren Verwendung als Fungizide |
| JP6121649B2 (ja) * | 2009-02-03 | 2017-04-26 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | 殺細菌剤としての硫黄含有複素芳香族酸類似体の使用 |
| BRPI1006006B1 (pt) | 2009-02-17 | 2018-05-22 | Bayer Intellectual Property Gmbh | Compostos, composição fungicida e método para o controle de fungos fitopatogênicos de culturas |
| EP2218717A1 (en) | 2009-02-17 | 2010-08-18 | Bayer CropScience AG | Fungicidal N-((HET)Arylethyl)thiocarboxamide derivatives |
| TW201031331A (en) | 2009-02-19 | 2010-09-01 | Bayer Cropscience Ag | Pesticide composition comprising a tetrazolyloxime derivative and a fungicide or an insecticide active substance |
| DE102009001469A1 (de) | 2009-03-11 | 2009-09-24 | Bayer Cropscience Ag | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| WO2010102758A2 (de) | 2009-03-11 | 2010-09-16 | Bayer Cropscience Ag | Halogenalkylmethylenoxy-phenyl-substituierte ketoenole |
| DE102010000662A1 (de) | 2009-03-18 | 2010-10-21 | Bayer Cropscience Ag | Aminopropylthiazol-Derivate als Fungizide |
| DE102009001681A1 (de) | 2009-03-20 | 2010-09-23 | Bayer Cropscience Ag | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| DE102009001732A1 (de) | 2009-03-23 | 2010-09-30 | Bayer Cropscience Ag | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| DE102009001730A1 (de) | 2009-03-23 | 2010-09-30 | Bayer Cropscience Ag | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| DE102009001728A1 (de) | 2009-03-23 | 2010-09-30 | Bayer Cropscience Ag | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| MX2011009830A (es) | 2009-03-25 | 2011-10-06 | Bayer Cropscience Ag | Combinaciones de principios activos con propiedades insecticidas y acaricidas. |
| US9012360B2 (en) | 2009-03-25 | 2015-04-21 | Bayer Intellectual Property Gmbh | Synergistic combinations of active ingredients |
| WO2010108505A1 (de) | 2009-03-25 | 2010-09-30 | Bayer Cropscience Ag | Wirkstoffkombinationen mit insektiziden und akariziden eigenschaften |
| CN102448305B (zh) | 2009-03-25 | 2015-04-01 | 拜尔农作物科学股份公司 | 具有杀昆虫和杀螨虫特性的活性成分结合物 |
| EP2232995A1 (de) | 2009-03-25 | 2010-09-29 | Bayer CropScience AG | Verfahren zur verbesserten Nutzung des Produktionspotentials transgener Pflanzen |
| EP2410847A1 (de) | 2009-03-25 | 2012-02-01 | Bayer CropScience AG | Wirkstoffkombinationen mit insektiziden und akariziden eigenschaften |
| EP2239331A1 (en) | 2009-04-07 | 2010-10-13 | Bayer CropScience AG | Method for improved utilization of the production potential of transgenic plants |
| BRPI1015543A8 (pt) | 2009-05-06 | 2016-05-24 | Bayer Cropscience Ag | Compostos de ciclopentanodiona e seu uso como inseticidas, acaricidas e/ou fungicidas. |
| EP2251331A1 (en) | 2009-05-15 | 2010-11-17 | Bayer CropScience AG | Fungicide pyrazole carboxamides derivatives |
| AR076839A1 (es) | 2009-05-15 | 2011-07-13 | Bayer Cropscience Ag | Derivados fungicidas de pirazol carboxamidas |
| CN102439013B (zh) | 2009-05-19 | 2015-03-18 | 拜尔农作物科学股份公司 | 螺杂环特窗酸衍生物 |
| EP2253617A1 (de) | 2009-05-20 | 2010-11-24 | Bayer CropScience AG | Halogen-substituierte Verbindungen als Pestizide |
| EP2255626A1 (de) | 2009-05-27 | 2010-12-01 | Bayer CropScience AG | Verwendung von Succinat Dehydrogenase Inhibitoren zur Steigerung der Resistenz von Pflanzen oder Pflanzenteilen gegenüber abiotischem Stress |
| BRPI1011983A2 (pt) | 2009-06-02 | 2015-09-22 | Bayer Cropscience Ag | utilização de inibidores de succinato desidrogenase para o controle sclerotinia ssp. |
| EP2264010A1 (de) | 2009-06-03 | 2010-12-22 | Bayer CropScience AG | Hetarylamidine |
| EP2264012A1 (de) | 2009-06-03 | 2010-12-22 | Bayer CropScience AG | Heteroarylamidine und deren Verwendung als Fungizide |
| EP2264011A1 (de) | 2009-06-03 | 2010-12-22 | Bayer CropScience AG | Heteroarylamidine und deren Verwendung als Fungizide |
| WO2010145789A1 (en) | 2009-06-18 | 2010-12-23 | Bayer Cropscience Ag | Propargyloxybenzamide derivatives |
| EP2272846A1 (de) | 2009-06-23 | 2011-01-12 | Bayer CropScience AG | Thiazolylpiperidin Derivate als Fungizide |
| EP2277868A1 (de) | 2009-06-24 | 2011-01-26 | Bayer CropScience AG | Phenyloxy(thio)phenylamidbenzoxa(thia)zole |
| EP2277870A1 (de) | 2009-06-24 | 2011-01-26 | Bayer CropScience AG | Substituierte Benzoxa(thia)zole |
| EP2277869A1 (de) | 2009-06-24 | 2011-01-26 | Bayer CropScience AG | Cycloalkylamidbenzoxa(thia)zole als Fungizide |
| EP2451785A2 (de) | 2009-07-08 | 2012-05-16 | Bayer CropScience AG | Substituierte phenyl(oxy/thio)alkanol-derivate |
| EP2451784A1 (de) | 2009-07-08 | 2012-05-16 | Bayer CropScience AG | Phenyl(oxy/thio)alkanol-derivate |
| AU2010272872B2 (en) | 2009-07-16 | 2014-08-28 | Bayer Intellectual Property Gmbh | Synergistic active substance combinations containing phenyl triazoles |
| WO2011006604A1 (de) | 2009-07-17 | 2011-01-20 | Bayer Cropscience Ag | Substituierte aminothiazole und deren verwendung als fungizide |
| WO2011015524A2 (en) | 2009-08-03 | 2011-02-10 | Bayer Cropscience Ag | Fungicide heterocycles derivatives |
| EP2292094A1 (en) | 2009-09-02 | 2011-03-09 | Bayer CropScience AG | Active compound combinations |
| AR077956A1 (es) | 2009-09-14 | 2011-10-05 | Bayer Cropscience Ag | Combinaciones de compuestos activos |
| WO2011032656A1 (de) | 2009-09-18 | 2011-03-24 | Bayer Cropscience Ag | 5-fluor-2-thio-substituierte pyrimidin-derivate |
| EP2308866A1 (de) | 2009-10-09 | 2011-04-13 | Bayer CropScience AG | Phenylpyri(mi)dinylpyrazole und ihre Verwendung als Fungizide |
| CN102695699B (zh) | 2009-10-16 | 2016-02-24 | 拜耳知识产权有限责任公司 | 作为杀菌剂的氨基丙烯酸酯 |
| WO2011051243A1 (en) | 2009-10-29 | 2011-05-05 | Bayer Cropscience Ag | Active compound combinations |
| JP5827626B2 (ja) | 2009-10-30 | 2015-12-02 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | ヘテロアリールピペリジン及びピペラジン誘導体 |
| WO2011051198A2 (de) | 2009-10-30 | 2011-05-05 | Bayer Cropscience Ag | Pyridin-derivate als pflanzenschutzmittel |
| UY33036A (es) | 2009-11-17 | 2011-06-30 | Bayer Cropscience Ag | Combinaciones de compuestos activos |
| EP2343280A1 (en) | 2009-12-10 | 2011-07-13 | Bayer CropScience AG | Fungicide quinoline derivatives |
| WO2011082941A1 (de) | 2009-12-16 | 2011-07-14 | Bayer Cropscience Ag | Benzylsubstituierte thiadiazolyloxyphenylamidiniumsalze als fungizide |
| ES2553620T3 (es) | 2009-12-21 | 2015-12-10 | Bayer Cropscience Ag | Bis(difluorometil)pirazoles como fungicidas |
| AR079545A1 (es) | 2009-12-21 | 2012-02-01 | Bayer Cropscience Ag | Tienilpiri(mi)dinilazol |
| TW201141381A (en) | 2009-12-28 | 2011-12-01 | Bayer Cropscience Ag | Fungicide hydroximoyl-tetrazole derivatives |
| BR112012012107B1 (pt) | 2009-12-28 | 2019-08-20 | Bayer Cropscience Ag | Composto, composição fungicida e método para controlar fungos fitopatogênico de culturas |
| CN105399666A (zh) | 2009-12-28 | 2016-03-16 | 拜尔农科股份公司 | 杀真菌剂肟基-杂环衍生物 |
| RS55986B1 (sr) | 2010-01-22 | 2017-09-29 | Bayer Ip Gmbh | Akaricidne i/ili insekticidne kombinacije aktivnih supstanci |
| BR112012020084B1 (pt) | 2010-02-10 | 2017-12-19 | Bayer Intellectual Property Gmbh | A process for the preparation of pesticides and / or herbicides and / or fungi and / or fungi and / or fungicides and / or fungicides and / or fungicides and / or fungicides. METHOD FOR INCREASING THE ACTION OF PESTICIDES AND / OR HERBICIDES AND / OR FUNGICIDES COMPREHENDING SUCH COMPOUNDS |
| EP2534147B1 (de) | 2010-02-10 | 2015-06-17 | Bayer Intellectual Property GmbH | Spiroheterocyclisch-substituierte tetramsäure-derivate |
| US8735560B1 (en) | 2010-03-02 | 2014-05-27 | Monsanto Technology Llc | Multiple domain lepidopteran active toxin proteins |
| WO2011107504A1 (de) | 2010-03-04 | 2011-09-09 | Bayer Cropscience Ag | Fluoralkyl- substituierte 2 -amidobenzimidazole und deren verwendung zur steigerung der stresstoleranz in pflanzen |
| UA108638C2 (uk) | 2010-03-04 | 2015-05-25 | Застосування солей імідів малеїнової кислоти для боротьби з фітопатогенними грибами | |
| BR112012023551A2 (pt) | 2010-03-18 | 2015-09-15 | Bayer Ip Gmbh | aril e hetaril sulfonamidas como agentes ativos contra estresse abiótico em plantas |
| WO2011117184A1 (de) | 2010-03-24 | 2011-09-29 | Bayer Cropscience Ag | Fludioxonil-derivate |
| WO2011124554A2 (de) | 2010-04-06 | 2011-10-13 | Bayer Cropscience Ag | Verwendung der 4-phenylbuttersäure und/oder ihrer salze zur steigerung der stresstoleranz in pflanzen |
| AR081810A1 (es) | 2010-04-07 | 2012-10-24 | Bayer Cropscience Ag | Piridinilpirazoles biciclicos |
| CN102933083B (zh) | 2010-04-09 | 2015-08-12 | 拜耳知识产权有限责任公司 | (1-氰基环丙基)苯基次膦酸或其酯的衍生物和/或其盐提高植物对非生物胁迫耐受性的用途 |
| ES2504095T3 (es) | 2010-04-14 | 2014-10-08 | Bayer Cropscience Ag | Derivados de ditiinopiridazinona como fungicidas |
| JP5764200B2 (ja) | 2010-04-14 | 2015-08-12 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | 活性化合物の組み合わせ |
| EP2557930A2 (en) | 2010-04-14 | 2013-02-20 | Bayer CropScience AG | Fungicidal combinations of dithiino-tetracarboxamide derivatives and inorganic salts |
| ES2483132T3 (es) | 2010-04-14 | 2014-08-05 | Bayer Cropscience Ag | Derivados de tienoditiina como fungicidas |
| EP2377867A1 (de) | 2010-04-14 | 2011-10-19 | Bayer CropScience AG | Dithiinopyridazinon-Derivate |
| EP2703397B1 (de) | 2010-04-14 | 2015-08-12 | Bayer Intellectual Property GmbH | Dithiin-derivate alz fungizide |
| KR20130100903A (ko) | 2010-04-28 | 2013-09-12 | 바이엘 크롭사이언스 아게 | 살진균제로서의 케토헤테로아릴피페리딘 및 케토헤테로아릴피페라진 유도체 |
| US20130116287A1 (en) | 2010-04-28 | 2013-05-09 | Christian Beier | Fungicide hydroximoyl-heterocycles derivatives |
| WO2011134911A2 (en) | 2010-04-28 | 2011-11-03 | Bayer Cropscience Ag | Fungicide hydroximoyl-tetrazole derivatives |
| JP2013525400A (ja) | 2010-04-28 | 2013-06-20 | バイエル・クロップサイエンス・アーゲー | 殺菌剤ヒドロキシモイル−複素環誘導体 |
| US8815775B2 (en) | 2010-05-18 | 2014-08-26 | Bayer Cropscience Ag | Bis(difluoromethyl)pyrazoles as fungicides |
| UA110704C2 (uk) | 2010-05-27 | 2016-02-10 | Баєр Інтеллекчуел Проперті Гмбх | Похідні гетероциклічних алканолів як фунгіциди |
| CA2800665A1 (en) | 2010-05-27 | 2011-12-01 | Carl Friedrich Nising | Heterocyclic alkanol derivatives as fungicides |
| JP5870093B2 (ja) | 2010-05-27 | 2016-02-24 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | 殺菌剤としてのヘテロ環式アルカノール誘導体 |
| WO2011147813A1 (de) | 2010-05-27 | 2011-12-01 | Bayer Cropscience Ag | Heterocyclische thiosubstituierte alkanolderivate als fungizide |
| CN103025723A (zh) | 2010-05-27 | 2013-04-03 | 拜尔农作物科学股份公司 | 作为杀真菌剂的吡啶基羧酸衍生物 |
| WO2011147811A1 (de) | 2010-05-27 | 2011-12-01 | Bayer Cropscience Ag | Heterocyclische alkanolderivate als fungizide |
| US8653114B2 (en) | 2010-06-03 | 2014-02-18 | Bayer Intellectual Property Gmbh | O-cyclopropylcyclohexyl-carboxanilides and their use as fungicides |
| CA2796191A1 (en) | 2010-06-03 | 2011-12-08 | Bayer Cropscience Ag | N-[(het)arylethyl)] pyrazole(thio)carboxamides and their heterosubstituted analogues |
| UA110703C2 (uk) | 2010-06-03 | 2016-02-10 | Байєр Кропсайнс Аг | Фунгіцидні похідні n-[(тризаміщений силіл)метил]-карбоксаміду |
| US8999956B2 (en) | 2010-06-03 | 2015-04-07 | Bayer Intellectual Property Gmbh | N-[(het)arylalkyl)] pyrazole(thio)carboxamides and their heterosubstituted analogues |
| US9574201B2 (en) | 2010-06-09 | 2017-02-21 | Bayer Cropscience Nv | Methods and means to modify a plant genome at a nucleotide sequence commonly used in plant genome engineering |
| AU2011264074B2 (en) | 2010-06-09 | 2015-01-22 | Bayer Cropscience Nv | Methods and means to modify a plant genome at a nucleotide sequence commonly used in plant genome engineering |
| WO2011161035A1 (de) | 2010-06-22 | 2011-12-29 | Bayer Cropscience Ag | 3-aryl-4-(2-thienylmethylen)-isoxazol-5(4h)-one als fungizide |
| WO2011161034A1 (de) | 2010-06-22 | 2011-12-29 | Bayer Cropscience Ag | 3-aryl-4-(2,6-dimethylbenzyliden)-isoxazol-5(4h)-one als fungizide |
| JP6215052B2 (ja) | 2010-06-28 | 2017-10-18 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | 有害生物防除剤としての複素環化合物 |
| CA2803083A1 (en) | 2010-07-20 | 2012-01-26 | Bayer Cropscience Ag | Benzocycloalkenes as antifungal agents |
| PL3058824T3 (pl) | 2010-08-05 | 2018-11-30 | Bayer Cropscience Ag | Kombinacje związków aktywnych zawierające protiokonazol i fluksapyroksad do zwalczania chorób kukurydzy |
| US20120122928A1 (en) | 2010-08-11 | 2012-05-17 | Bayer Cropscience Ag | Heteroarylpiperidine and -Piperazine Derivatives as Fungicides |
| EP2423210A1 (de) | 2010-08-25 | 2012-02-29 | Bayer CropScience AG | Heteroarylpiperidin- und -piperazinderivate als Fungizide |
| US8759527B2 (en) | 2010-08-25 | 2014-06-24 | Bayer Cropscience Ag | Heteroarylpiperidine and -piperazine derivatives as fungicides |
| NZ607381A (en) | 2010-08-26 | 2014-10-31 | Bayer Ip Gmbh | 5-iodo-triazole derivatives |
| WO2012028587A1 (de) | 2010-09-03 | 2012-03-08 | Bayer Cropscience Ag | Dithiin-tetra (thio) carboximide zur bekämpfung phytopathogener pilze |
| RU2013114710A (ru) | 2010-09-03 | 2014-10-10 | Байер Интеллектуэль Проперти Гмбх | Замещенные конденсированные пиримидиноны и дигидропиримидиноны |
| JP2012082186A (ja) | 2010-09-15 | 2012-04-26 | Bayer Cropscience Ag | 殺虫性アリールピロリジン類 |
| JP2012062267A (ja) | 2010-09-15 | 2012-03-29 | Bayer Cropscience Ag | 殺虫性ピロリンｎ−オキサイド誘導体 |
| EP2460406A1 (en) | 2010-12-01 | 2012-06-06 | Bayer CropScience AG | Use of fluopyram for controlling nematodes in nematode resistant crops |
| WO2012038480A2 (en) | 2010-09-22 | 2012-03-29 | Bayer Cropscience Ag | Use of biological or chemical control agents for controlling insects and nematodes in resistant crops |
| WO2012045798A1 (en) | 2010-10-07 | 2012-04-12 | Bayer Cropscience Ag | Fungicide composition comprising a tetrazolyloxime derivative and a thiazolylpiperidine derivative |
| WO2012045726A2 (en) | 2010-10-07 | 2012-04-12 | Bayer Cropscience Ag | 5-heteroarylimino-1,2,3-dithiazoles |
| CA2815114A1 (en) | 2010-10-21 | 2012-04-26 | Juergen Benting | 1-(heterocyclic carbonyl) piperidines |
| UA107865C2 (ru) | 2010-10-21 | 2015-02-25 | Байєр Інтелекчуал Проперті Гмбх | Гетероциклические карбоксамиды |
| US20140005224A1 (en) | 2010-10-27 | 2014-01-02 | Bayer Intellectual Property Gmbh | Heteroaryl piperidine and heteroaryl piperazine derivatives as fungicides |
| WO2012059497A1 (en) | 2010-11-02 | 2012-05-10 | Bayer Cropscience Ag | N-hetarylmethyl pyrazolylcarboxamides |
| WO2012062749A1 (de) | 2010-11-12 | 2012-05-18 | Bayer Cropscience Ag | Benzimidazolidinone verwendbar als fungizide |
| AR083874A1 (es) | 2010-11-15 | 2013-03-27 | Bayer Cropscience Ag | 5-halogenopirazol(tio)carboxamidas |
| WO2012065905A1 (en) | 2010-11-15 | 2012-05-24 | Bayer Cropscience Ag | Cyanoenamines and their use as fungicides |
| WO2012065947A1 (en) | 2010-11-15 | 2012-05-24 | Bayer Cropscience Ag | 5-halogenopyrazolecarboxamides |
| BR112013012080A2 (pt) | 2010-11-15 | 2016-07-19 | Bayer Ip Gmbh | n-aril pirazol (tio) carboxamidas |
| ES2643128T3 (es) | 2010-11-15 | 2017-11-21 | Bayer Intellectual Property Gmbh | Cianoenaminas y su uso como fungicidas |
| EP2454939A1 (en) | 2010-11-18 | 2012-05-23 | Bayer CropScience AG | Post-harvest treatment |
| EA023763B1 (ru) | 2010-11-30 | 2016-07-29 | Байер Интеллектчуал Проперти Гмбх | Производные пиримидина и их применение в качестве пестицидов |
| EP3372081A3 (en) | 2010-12-01 | 2018-10-24 | Bayer CropScience Aktiengesellschaft | Use of fluopyram for controlling nematodes in crops |
| EP2460407A1 (de) | 2010-12-01 | 2012-06-06 | Bayer CropScience AG | Wirkstoffkombinationen umfassend Pyridylethylbenzamide und weitere Wirkstoffe |
| US20130289077A1 (en) | 2010-12-29 | 2013-10-31 | Juergen Benting | Fungicide hydroximoyl-tetrazole derivatives |
| EP2474542A1 (en) | 2010-12-29 | 2012-07-11 | Bayer CropScience AG | Fungicide hydroximoyl-tetrazole derivatives |
| EP2471363A1 (de) | 2010-12-30 | 2012-07-04 | Bayer CropScience AG | Verwendung von Aryl-, Heteroaryl- und Benzylsulfonamidocarbonsäuren, -carbonsäureestern, -carbonsäureamiden und -carbonitrilen oder deren Salze zur Steigerung der Stresstoleranz in Pflanzen |
| WO2012088645A1 (en) | 2010-12-31 | 2012-07-05 | Bayer Cropscience Ag | Method for improving plant quality |
| MX352307B (es) | 2011-02-01 | 2017-11-17 | Bayer Ip Gmbh | Derivados de heteroarilpiperidina y -piperazina como fungicidas. |
| KR20140018879A (ko) | 2011-02-15 | 2014-02-13 | 바이엘 인텔렉쳐 프로퍼티 게엠베하 | 디티이노 테트라카복사미드 살진균제 및 제초제, 약해완화제 또는 식물 성장 조절제를 함유하는 상승적 배합물 |
| EP2675789A1 (de) | 2011-02-17 | 2013-12-25 | Bayer Intellectual Property GmbH | Substituierte 3-(biphenyl-3-yl)-8,8-difluor-4-hydroxy-1-azaspiro[4.5]dec-3-en-2-one zur therapie und halogensubstituierte spirocyclische ketoenole |
| EP2494867A1 (de) | 2011-03-01 | 2012-09-05 | Bayer CropScience AG | Halogen-substituierte Verbindungen in Kombination mit Fungiziden |
| MX344895B (es) | 2011-03-01 | 2017-01-09 | Bayer Ip Gmbh | 2-aciloxipirrolin-4-onas. |
| CN102181460B (zh) * | 2011-03-09 | 2012-09-19 | 黑龙江大学 | 玉米螟谷胱甘肽-S-转移酶基因的RNAi干扰工程菌的构建方法 |
| CA2823999C (en) | 2011-03-10 | 2020-03-24 | Bayer Intellectual Property Gmbh | Use of lipochito-oligosaccharide compounds for safeguarding seed safety of treated seeds |
| EP2499911A1 (en) | 2011-03-11 | 2012-09-19 | Bayer Cropscience AG | Active compound combinations comprising fenhexamid |
| BR112013023502A2 (pt) | 2011-03-14 | 2016-08-02 | Bayer Ip Gmbh | composto fórmula (i), composição fungicida, método para o controle de fungos fitopatogênicos de culturas, utilização dos compostos de fórmula (i) e processo para a produção das composições |
| EP2502495A1 (en) | 2011-03-16 | 2012-09-26 | Bayer CropScience AG | Use of a dithiino-tetracarboxamide for the protection of harvested products against phytopathogenic fungi |
| ES2564972T3 (es) | 2011-03-18 | 2016-03-30 | Bayer Intellectual Property Gmbh | Derivados de N-(3-carbamoilfenil)-1H-pirazol-5-carboxamida y su uso para combatir parásitos animales |
| IN2013CN07575A (zh) | 2011-03-23 | 2015-07-31 | Bayer Ip Gmbh | |
| WO2012130686A2 (en) | 2011-03-25 | 2012-10-04 | Bayer Cropscience Ag | Active compound combinations |
| CA2831704C (en) | 2011-03-31 | 2019-10-01 | Bayer Intellectual Property Gmbh | Herbicidally and fungicidally active 3-phenylisoxazoline-5-carboxamides and 3-phenylisoxazoline-5-thioamides |
| US20140051575A1 (en) | 2011-04-08 | 2014-02-20 | Juergen Benting | Fungicide hydroximoyl-tetrazole derivatives |
| AR085587A1 (es) | 2011-04-13 | 2013-10-09 | Bayer Cropscience Ag | Combinaciones de compuestos activos |
| AR085588A1 (es) | 2011-04-13 | 2013-10-09 | Bayer Cropscience Ag | Combinaciones de compuestos activos |
| AR085568A1 (es) | 2011-04-15 | 2013-10-09 | Bayer Cropscience Ag | 5-(biciclo[4.1.0]hept-3-en-2-il)-penta-2,4-dienos y 5-(biciclo[4.1.0]hept-3-en-2-il)-pent-2-en-4-inos sustituidos como principios activos contra el estres abiotico de las plantas |
| AR085585A1 (es) | 2011-04-15 | 2013-10-09 | Bayer Cropscience Ag | Vinil- y alquinilciclohexanoles sustituidos como principios activos contra estres abiotico de plantas |
| AR090010A1 (es) | 2011-04-15 | 2014-10-15 | Bayer Cropscience Ag | 5-(ciclohex-2-en-1-il)-penta-2,4-dienos y 5-(ciclohex-2-en-1-il)-pent-2-en-4-inos sustituidos como principios activos contra el estres abiotico de las plantas, usos y metodos de tratamiento |
| EP2511255A1 (de) | 2011-04-15 | 2012-10-17 | Bayer CropScience AG | Substituierte Prop-2-in-1-ol- und Prop-2-en-1-ol-Derivate |
| EP2510787A1 (en) | 2011-04-15 | 2012-10-17 | Bayer Cropscience AG | Propenoates as fungicides |
| DK2997825T3 (en) | 2011-04-22 | 2019-03-11 | Bayer Ip Gmbh | COMPOSITIONS OF ACTIVE COMPOUNDS CONTAINING A (THIO) CARBOXAMIDE DERIVATIVE AND A FUNGICID COMPOUND |
| EP2524601A1 (en) | 2011-05-17 | 2012-11-21 | Bayer CropScience AG | Active compound combinations comprising a phosphorous acid derivative and cyazofamid |
| EP2524600A1 (en) | 2011-05-17 | 2012-11-21 | Bayer CropScience AG | Active compound combinations comprising phosphorous acid or a derivative thereof and Tebuconazole or Myclobutanil |
| EP2524598A1 (en) | 2011-05-17 | 2012-11-21 | Bayer CropScience AG | Active compound combinations comprising dithianon |
| EP2524599A1 (en) | 2011-05-17 | 2012-11-21 | Bayer CropScience AG | Active compound combinations |
| HUE037039T2 (hu) | 2011-05-17 | 2018-08-28 | Bayer Ip Gmbh | Aktív vegyület kombinációk |
| TR201802544T4 (tr) | 2011-06-06 | 2018-03-21 | Bayer Cropscience Nv | Önceden seçilmiş bir bölgede bir bitki genomunu modifiye için yöntemler ve araçlar. |
| EP2532233A1 (en) | 2011-06-07 | 2012-12-12 | Bayer CropScience AG | Active compound combinations |
| US9241493B2 (en) | 2011-06-14 | 2016-01-26 | Bayer Intellectual Property Gmbh | Use of an enaminocarbonyl compound in combination with a biological control agent |
| AR086992A1 (es) | 2011-06-20 | 2014-02-05 | Bayer Ip Gmbh | Tienilpiri(mi)dinilpirazoles |
| EP2540165A1 (en) | 2011-06-30 | 2013-01-02 | Bayer CropScience AG | Use of a halogenated pesticide in combination with a biological pest control agent |
| EP2729007A1 (de) | 2011-07-04 | 2014-05-14 | Bayer Intellectual Property GmbH | Verwendung substituierter isochinolinone, isochinolindione, isochinolintrione und dihydroisochinolinone oder jeweils deren salze als wirkstoffe gegen abiotischen pflanzenstress |
| BR102012019434B1 (pt) * | 2011-07-26 | 2021-11-09 | Dow Agrosciences Llc | Métodos de controle de pestes, de insetos, molécula e sequência de dna diagnóstica para o evento de soja 9582.814.19.1 |
| AU2012288866B2 (en) | 2011-07-27 | 2016-06-16 | Bayer Cropscience Aktiengesellschaft | Seed dressing for controlling phytopathogenic fungi |
| WO2013020985A1 (en) | 2011-08-10 | 2013-02-14 | Bayer Intellectual Property Gmbh | Active compound combinations comprising specific tetramic acid derivatives |
| CA2855948C (en) | 2011-08-11 | 2020-07-28 | Bayer Cropscience Ag | 1,2,4-triazolyl-substituted keto-enols |
| BR112014002988A2 (pt) | 2011-08-12 | 2017-03-01 | Bayer Cropscience Nv | expressão específica de célula de proteção de transgenes em algodão |
| JP2014524455A (ja) | 2011-08-22 | 2014-09-22 | バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー | 殺真菌性ヒドロキシモイル−テトラゾール誘導体 |
| BR112014003919A2 (pt) | 2011-08-22 | 2017-03-14 | Bayer Cropscience Ag | métodos e meios para modificar um genoma de planta |
| EP2561759A1 (en) | 2011-08-26 | 2013-02-27 | Bayer Cropscience AG | Fluoroalkyl-substituted 2-amidobenzimidazoles and their effect on plant growth |
| JP2014530173A (ja) | 2011-09-09 | 2014-11-17 | バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー | 植物の収量を改善するためのアシル−ホモセリンラクトン誘導体 |
| US9090600B2 (en) | 2011-09-12 | 2015-07-28 | Bayer Intellectual Property Gmbh | Fungicidal 4-substituted-3-{phenyl[(heterocyclylmethoxy)imino]methyl}-1,2,4-oxadizol-5(4H)-one derivatives |
| US10004232B2 (en) | 2011-09-15 | 2018-06-26 | Bayer Intellectual Property Gmbh | Piperidine pyrazoles as fungicides |
| AR087874A1 (es) | 2011-09-16 | 2014-04-23 | Bayer Ip Gmbh | Uso de acilsulfonamidas para mejorar el rendimiento de las plantas |
| CN103781352A (zh) | 2011-09-16 | 2014-05-07 | 拜耳知识产权有限责任公司 | 苯基吡唑啉-3-甲酸酯类用于提高植物产量的用途 |
| US20140378306A1 (en) | 2011-09-16 | 2014-12-25 | Bayer Intellectual Property Gmbh | Use of 5-phenyl- or 5-benzyl-2 isoxazoline-3 carboxylates for improving plant yield |
| JP2014527973A (ja) | 2011-09-23 | 2014-10-23 | バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー | 非生物的な植物ストレスに対する作用剤としての４−置換１−フェニルピラゾール−３−カルボン酸誘導体の使用 |
| AU2012320554B2 (en) | 2011-10-04 | 2017-11-09 | Bayer Intellectual Property Gmbh | RNAi for the control of fungi and oomycetes by inhibiting saccharopine dehydrogenase gene |
| WO2013050324A1 (de) | 2011-10-06 | 2013-04-11 | Bayer Intellectual Property Gmbh | Abiotischen pflanzenstress-reduzierende kombination enthaltend 4- phenylbuttersäure (4-pba) oder eines ihrer salze (komponente (a)) und eine oder mehrere ausgewählte weitere agronomisch wirksame verbindungen (komponente(n) (b) |
| MX351460B (es) | 2011-10-06 | 2017-10-16 | Bayer Ip Gmbh | Heterociclilpiri(mi)dinilpirazol. |
| CN104039785A (zh) | 2011-10-06 | 2014-09-10 | 拜耳知识产权有限责任公司 | 作为杀真菌剂的杂环基吡(嘧)啶基吡唑 |
| MX2014005976A (es) | 2011-11-21 | 2014-08-27 | Bayer Ip Gmbh | Derivados de n-[(silil trisustituido)metil]-carboxamida fungicidas. |
| EP2782447A1 (de) | 2011-11-25 | 2014-10-01 | Bayer Intellectual Property GmbH | 2-iod-imidazol-derivate |
| CA2856711A1 (en) | 2011-11-25 | 2013-05-30 | Bayer Intellectual Property Gmbh | Novel heterocyclic alkanol-derivatives |
| CN105906567B (zh) | 2011-11-30 | 2019-01-22 | 拜耳知识产权有限责任公司 | 杀真菌的n-二环烷基和n-三环烷基(硫代)羧酰胺衍生物 |
| EP2601839A1 (en) | 2011-12-08 | 2013-06-12 | Bayer CropScience AG | Synergisitic fungicidal combinations containing phosphorous acid derivative and zoxamide |
| BR112014015002A2 (pt) | 2011-12-19 | 2017-06-13 | Bayer Cropscience Ag | uso de derivados de diamida de ácido antranílico para o controle de pragas em culturas transgênicas |
| EP2606732A1 (en) | 2011-12-19 | 2013-06-26 | Bayer CropScience AG | Use of an anthranilic diamide derivatives with heteroaromatic and heterocyclic substituents in combination with a biological control agent |
| JP6062958B2 (ja) | 2011-12-20 | 2017-01-18 | バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングＢａｙｅｒ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ ＧｍｂＨ | 新規殺虫性芳香族アミド類 |
| ES2675506T3 (es) | 2011-12-27 | 2018-07-11 | Bayer Cropscience Aktiengesellschaft | Derivados de heteroarilpiperidina y de heteroarilpiperazina como fungicidas |
| BR112014015993A8 (pt) | 2011-12-29 | 2017-07-04 | Bayer Ip Gmbh | composto, composição, método para o controle dos fungos, utilização dos compostos e processo para a produção das composições |
| KR102028903B1 (ko) | 2011-12-29 | 2019-10-07 | 바이엘 인텔렉쳐 프로퍼티 게엠베하 | 살진균 3-[(피리딘-2-일메톡시이미노)(페닐)메틸]-2-치환-1,2,4-옥사디아졸-5(2h)-온 유도체 |
| EP2806739A1 (en) | 2012-01-25 | 2014-12-03 | Bayer Intellectual Property GmbH | Active compound combinations containing fluopyram and biological control agent |
| CA2862166A1 (en) | 2012-01-25 | 2013-08-01 | Bayer Intellectual Property Gmbh | Active compounds combination containing fluopyram bacillus and biologically control agent |
| EP2622961A1 (en) | 2012-02-02 | 2013-08-07 | Bayer CropScience AG | Acive compound combinations |
| PT2816897T (pt) | 2012-02-22 | 2018-04-02 | Bayer Cropscience Ag | Utilização de fluopiram para controlar doenças da madeira em uvas |
| EP2819518B1 (en) | 2012-02-27 | 2017-09-06 | Bayer Intellectual Property GmbH | Active compound combinations containing a thiazoylisoxazoline and a fungicide |
| EP2825532A1 (en) | 2012-03-14 | 2015-01-21 | Bayer Intellectual Property GmbH | Pesticidal arylpyrrolidines |
| WO2013139949A1 (en) | 2012-03-23 | 2013-09-26 | Bayer Intellectual Property Gmbh | Compositions comprising a strigolactame compound for enhanced plant growth and yield |
| WO2013153143A1 (en) | 2012-04-12 | 2013-10-17 | Bayer Cropscience Ag | N-acyl- 2 - (cyclo) alkylpyrrolidines and piperidines useful as fungicides |
| CN104428294B (zh) | 2012-04-20 | 2017-07-14 | 拜尔农科股份公司 | N‑环烷基‑n‑[(杂环基苯基)亚甲基]‑(硫代)羧酰胺衍生物 |
| US20150080337A1 (en) | 2012-04-20 | 2015-03-19 | Bayer Cropscience | N-cycloalkyl-n-[(trisubstitutedsilylphenyl)methylene]-(thio)carboxamide derivatives |
| EP2841581B2 (en) | 2012-04-23 | 2023-03-08 | BASF Agricultural Solutions Seed US LLC | Targeted genome engineering in plants |
| EP2662361A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | Pyrazol indanyl carboxamides |
| EP2847170B1 (en) | 2012-05-09 | 2017-11-08 | Bayer CropScience AG | Pyrazole indanyl carboxamides |
| WO2013167544A1 (en) | 2012-05-09 | 2013-11-14 | Bayer Cropscience Ag | 5-halogenopyrazole indanyl carboxamides |
| EP2662364A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | Pyrazole tetrahydronaphthyl carboxamides |
| EP2662360A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | 5-Halogenopyrazole indanyl carboxamides |
| EP2662362A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | Pyrazole indanyl carboxamides |
| EP2662370A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | 5-Halogenopyrazole benzofuranyl carboxamides |
| EP2662363A1 (en) | 2012-05-09 | 2013-11-13 | Bayer CropScience AG | 5-Halogenopyrazole biphenylcarboxamides |
| AR091104A1 (es) | 2012-05-22 | 2015-01-14 | Bayer Cropscience Ag | Combinaciones de compuestos activos que comprenden un derivado lipo-quitooligosacarido y un compuesto nematicida, insecticida o fungicida |
| EP3318128A3 (en) | 2012-05-30 | 2018-06-27 | Bayer CropScience Aktiengesellschaft | Composition comprising a biological control agent and a fungicide |
| CN104837350B (zh) | 2012-05-30 | 2018-08-03 | 拜尔农作物科学股份公司 | 包含生物防治剂和杀真菌剂的组合物作为杀真菌剂的用途 |
| EP3292764A3 (en) | 2012-05-30 | 2018-04-25 | Bayer CropScience Aktiengesellschaft | Composition comprising a biological control agent and a fungicide selected from inhibitors of the respiratory chain at complex iii |
| US9386773B2 (en) | 2012-05-30 | 2016-07-12 | Bayer Cropscience Ag | Compositions comprising a biological control agent and a fungicide from the group consisting of inhibitors of the respiratory chain at complex I or II |
| EP3360418A1 (en) | 2012-05-30 | 2018-08-15 | Bayer CropScience Aktiengesellschaft | Composition comprising a biological control agent and a fungicide |
| AU2013269723B2 (en) | 2012-05-30 | 2016-12-15 | Bayer Cropscience Ag | Composition comprising a biological control agent and a fungicide |
| NZ701724A (en) | 2012-05-30 | 2016-11-25 | Bayer Cropscience Ag | Compositions comprising a biological control agent and an insecticide |
| ES2698951T3 (es) | 2012-05-30 | 2019-02-06 | Bayer Cropscience Ag | Composiciones que comprenden un agente de control biológico y un insecticida |
| EP2871958A1 (en) | 2012-07-11 | 2015-05-20 | Bayer CropScience AG | Use of fungicidal combinations for increasing the tolerance of a plant towards abiotic stress |
| CN104602520A (zh) | 2012-07-31 | 2015-05-06 | 拜尔农作物科学股份公司 | 包括杀虫萜烯混合物和杀虫剂的组合物 |
| CA2883574A1 (en) | 2012-09-05 | 2014-03-13 | Bayer Cropscience Ag | Use of substituted 2-amidobenzimidazoles, 2-amidobenzoxazoles and 2-amidobenzothiazoles or salts thereof as active substances against abiotic plant stress |
| JP6285937B2 (ja) | 2012-09-25 | 2018-02-28 | バイエル・クロップサイエンス・アクチェンゲゼルシャフト | 除草性および殺真菌性の５−オキシ置換３−フェニルイソオキサゾリン−５−カルボキサミドおよび５−オキシ置換３−フェニルイソオキサゾリン−５−チオアミド |
| DE102012219029A1 (de) | 2012-10-18 | 2014-04-24 | Bayer Cropscience Ag | Verwendung von Dithiin-tetracarboximiden zum Bekämpfen von neuer Blattfallkrankheit Marssonia coronaria |
| US9668480B2 (en) | 2012-10-19 | 2017-06-06 | Bayer Cropscience Ag | Method for treating plants against fungi resistant to fungicides using carboxamide or thiocarboxamide derivatives |
| PL2908642T3 (pl) | 2012-10-19 | 2022-06-13 | Bayer Cropscience Ag | Sposób wzmacniania tolerancji roślin na stres abiotyczny z zastosowaniem pochodnych karboksyamidowych lub tiokarboksyamidowych |
| CN105357967B (zh) | 2012-10-19 | 2019-02-19 | 拜尔农科股份公司 | 使用羧酰胺衍生物促进植物生长的方法 |
| US9801374B2 (en) | 2012-10-19 | 2017-10-31 | Bayer Cropscience Ag | Active compound combinations comprising carboxamide derivatives |
| MX355153B (es) | 2012-10-19 | 2018-04-06 | Bayer Cropscience Ag | Combinaciones de compuestos activos que comprenden derivados de carboxamida y un agente de control biologico. |
| CN102993282B (zh) * | 2012-11-19 | 2014-10-15 | 北京大北农科技集团股份有限公司 | 杀虫蛋白质、其编码基因及用途 |
| CN102981001B (zh) * | 2012-11-22 | 2014-11-12 | 中国农业科学院植物保护研究所 | 一种评价棉铃虫抗性治理效果的方法 |
| WO2014079957A1 (de) | 2012-11-23 | 2014-05-30 | Bayer Cropscience Ag | Selektive inhibition der ethylensignaltransduktion |
| EP2735231A1 (en) | 2012-11-23 | 2014-05-28 | Bayer CropScience AG | Active compound combinations |
| BR112015012519A2 (pt) | 2012-11-30 | 2017-07-11 | Bayer Cropscience Ag | misturas ternárias fungicidas e pesticidas |
| WO2014082950A1 (en) | 2012-11-30 | 2014-06-05 | Bayer Cropscience Ag | Ternary fungicidal mixtures |
| UA117820C2 (uk) | 2012-11-30 | 2018-10-10 | Байєр Кропсайєнс Акцієнгезелльшафт | Подвійна фунгіцидна або пестицидна суміш |
| BR112015012473A2 (pt) | 2012-11-30 | 2017-07-11 | Bayer Cropscience Ag | misturas binárias pesticidas e fungicidas |
| WO2014083088A2 (en) | 2012-11-30 | 2014-06-05 | Bayer Cropscience Ag | Binary fungicidal mixtures |
| WO2014086753A2 (en) | 2012-12-03 | 2014-06-12 | Bayer Cropscience Ag | Composition comprising biological control agents |
| MX2015006500A (es) | 2012-12-03 | 2015-08-14 | Bayer Cropsciense Ag | Composicion que comprende un agente de control biologico y un fungicida. |
| US9867377B2 (en) | 2012-12-03 | 2018-01-16 | Bayer Cropscience Ag | Composition comprising a biological control agent and an insecticide |
| CN105163589A (zh) | 2012-12-03 | 2015-12-16 | 拜耳作物科学股份公司 | 包括生物防治剂及杀真菌剂的组合物 |
| CN105451562A (zh) | 2012-12-03 | 2016-03-30 | 拜耳作物科学股份公司 | 包含生物防治剂和杀真菌剂的组合物 |
| BR112015012781A2 (pt) | 2012-12-03 | 2018-06-26 | Bayer Cropscience Ag | composição compreendendo agentes de controle biológico |
| BR112015012763B1 (pt) | 2012-12-03 | 2020-05-12 | Bayer Cropscience Ag | Composição, semente revestida com uma composição, uso da composição, kit de componentes e método para reduzir danos globais em plantas e controlar nematodes e insetos |
| US20150289518A1 (en) | 2012-12-03 | 2015-10-15 | Bayer Cropscience Ag | Composition comprising a biological control agent and an insecticide |
| EP2740356A1 (de) | 2012-12-05 | 2014-06-11 | Bayer CropScience AG | Substituierte (2Z)-5(1-Hydroxycyclohexyl)pent-2-en-4-insäure-Derivate |
| EP2740720A1 (de) | 2012-12-05 | 2014-06-11 | Bayer CropScience AG | Substituierte bicyclische- und tricyclische Pent-2-en-4-insäure -Derivate und ihre Verwendung zur Steigerung der Stresstoleranz in Pflanzen |
| EP2928296A1 (de) | 2012-12-05 | 2015-10-14 | Bayer CropScience AG | Verwendung substituierter 1-(arylethinyl)-, 1-(heteroarylethinyl)-, 1-(heterocyclylethinyl)- und 1-(cyloalkenylethinyl)-cyclohexanole als wirkstoffe gegen abiotischen pflanzenstress |
| WO2014090765A1 (en) | 2012-12-12 | 2014-06-19 | Bayer Cropscience Ag | Use of 1-[2-fluoro-4-methyl-5-(2,2,2-trifluoroethylsulfinyl)phenyl]-5-amino-3-trifluoromethyl)-1 h-1,2,4 tfia zole for controlling nematodes in nematode-resistant crops |
| AR093996A1 (es) | 2012-12-18 | 2015-07-01 | Bayer Cropscience Ag | Combinaciones bactericidas y fungicidas binarias |
| WO2014095677A1 (en) | 2012-12-19 | 2014-06-26 | Bayer Cropscience Ag | Difluoromethyl-nicotinic- tetrahydronaphtyl carboxamides |
| MY172725A (en) | 2013-02-06 | 2019-12-11 | Vetoquinol Sa | Halogen-substituted pyrazol derivatives as pest-control agents |
| BR112015018419A2 (pt) | 2013-02-11 | 2017-07-18 | Bayer Cropscience Lp | composições compreendendo um agente de controle biológico à base de streptomyces e um outro agente de controle biológico |
| BR112015018113A2 (pt) | 2013-02-11 | 2017-07-18 | Bayer Cropscience Lp | composições compreendendo um agente de controle biológico baseado em streptomyces e um fungicida |
| US20150366199A1 (en) | 2013-02-11 | 2015-12-24 | Bayer Cropscience Lp | Compositions comprising gougerotin and an insecticide |
| JP2016515100A (ja) | 2013-03-07 | 2016-05-26 | バイエル・クロップサイエンス・アクチェンゲゼルシャフト | 殺菌性３−｛フェニル［（ヘテロシクリルメトキシ）イミノ］メチル｝−ヘテロ環誘導体 |
| US10316327B2 (en) | 2013-03-15 | 2019-06-11 | Yi Li | Genetically modified plants that are insect-resistant and/or rot resistant |
| CA2908403A1 (en) | 2013-04-02 | 2014-10-09 | Bayer Cropscience Nv | Targeted genome engineering in eukaryotes |
| US9550752B2 (en) | 2013-04-12 | 2017-01-24 | Bayer Cropscience Aktiengesellschaft | Triazolinthione derivatives |
| MX2015014365A (es) | 2013-04-12 | 2015-12-07 | Bayer Cropscience Ag | Derivados de triazol novedosos. |
| US9554573B2 (en) | 2013-04-19 | 2017-01-31 | Bayer Cropscience Aktiengesellschaft | Binary insecticidal or pesticidal mixture |
| US10526378B2 (en) | 2013-04-19 | 2020-01-07 | Agresearch Limited | Methods and materials for encapsulating proteins |
| CA2909725A1 (en) | 2013-04-19 | 2014-10-23 | Bayer Cropscience Aktiengesellschaft | Method for improved utilization of the production potential of transgenic plants |
| WO2014177514A1 (en) | 2013-04-30 | 2014-11-06 | Bayer Cropscience Ag | Nematicidal n-substituted phenethylcarboxamides |
| TW201507722A (zh) | 2013-04-30 | 2015-03-01 | Bayer Cropscience Ag | 做為殺線蟲劑及殺體內寄生蟲劑的ｎ－（２－鹵素－２－苯乙基）－羧醯胺類 |
| EP2801575A1 (en) | 2013-05-07 | 2014-11-12 | Bayer CropScience AG | Heteroaryldihydropyridine derivatives as fungicides |
| EP3296403A1 (en) | 2013-06-14 | 2018-03-21 | Monsanto Technology LLC | Soybean transgenic event mon87751 and methods for detection and use thereof |
| CN105636939B (zh) | 2013-06-26 | 2018-08-31 | 拜耳作物科学股份公司 | N-环烷基-n-[(二环基苯基)亚甲基]-(硫代)甲酰胺衍生物 |
| AR096827A1 (es) | 2013-07-09 | 2016-02-03 | Bayer Cropscience Ag | Uso de piridoncarboxamidas seleccionadas o sus sales como ingredientes activos contra estrés abiótico en plantas |
| MX2016003405A (es) | 2013-09-24 | 2016-10-28 | Bayer Cropscience Nv | Hetero-transglicosilasa novedosa y usos de la misma. |
| CN103718895B (zh) * | 2013-11-18 | 2016-05-18 | 北京大北农科技集团股份有限公司 | 控制害虫的方法 |
| CN103718896B (zh) * | 2013-11-18 | 2016-02-10 | 北京大北农科技集团股份有限公司 | 控制害虫的方法 |
| US9970926B1 (en) | 2013-11-22 | 2018-05-15 | Monsanto Technology Llc | Bacillus thuringiensis toxin receptors and uses thereof |
| CN105873907B (zh) | 2013-12-05 | 2019-03-12 | 拜耳作物科学股份公司 | N-环烷基-n-{[2-(1-取代的环烷基)苯基]亚甲基}-(硫代)甲酰胺衍生物 |
| AU2014359208B2 (en) | 2013-12-05 | 2018-10-04 | Bayer Cropscience Aktiengesellschaft | N-cycloalkyl-N-{[2-(1-substitutedcycloalkyl)phenyl]methylene}-(thio)carboxamide derivatives |
| EP3719129A1 (en) * | 2014-03-12 | 2020-10-07 | The University of Sydney | Rna production in higher plants |
| WO2015160619A1 (en) | 2014-04-16 | 2015-10-22 | Bayer Cropscience Lp | Compositions comprising ningnanmycin and a fungicide |
| WO2015160620A1 (en) | 2014-04-16 | 2015-10-22 | Bayer Cropscience Lp | Compositions comprising ningnanmycin and an insecticide |
| WO2015160618A1 (en) | 2014-04-16 | 2015-10-22 | Bayer Cropscience Lp | Compositions comprising ningnanmycin and a biological control agent |
| AR101214A1 (es) | 2014-07-22 | 2016-11-30 | Bayer Cropscience Ag | Ciano-cicloalquilpenta-2,4-dienos, ciano-cicloalquilpent-2-en-4-inas, ciano-heterociclilpenta-2,4-dienos y ciano-heterociclilpent-2-en-4-inas sustituidos como principios activos contra el estrés abiótico de plantas |
| WO2016061392A1 (en) | 2014-10-16 | 2016-04-21 | Monsanto Technology Llc | Proteins toxic or inhibitory to lepidopteran insects |
| UA123036C2 (uk) | 2014-10-16 | 2021-02-03 | Монсанто Текнолоджі Ллс | Сконструйований інсектицидний білок, який має активність проти лускокрилих |
| US10487123B2 (en) | 2014-10-16 | 2019-11-26 | Monsanto Technology Llc | Chimeric insecticidal proteins toxic or inhibitory to lepidopteran pests |
| BR122020006739B1 (pt) | 2014-11-20 | 2024-01-23 | Monsanto Technology Llc | Cassete de expressão, célula hospedeira bacteriana, composição inibidora de inseto, método para controlar uma praga de espécie coleoptera, e vetor recombinante |
| CN107109418B (zh) * | 2014-12-12 | 2021-07-16 | 先正达参股股份有限公司 | 用于控制植物有害生物的组合物和方法 |
| AR103024A1 (es) | 2014-12-18 | 2017-04-12 | Bayer Cropscience Ag | Piridoncarboxamidas seleccionadas o sus sales como sustancias activas contra estrés abiótico de las plantas |
| CN104522033B (zh) * | 2014-12-22 | 2016-09-14 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| CN104628835B (zh) * | 2015-01-29 | 2017-11-21 | 中国农业科学院油料作物研究所 | 一种与植物耐虫性相关蛋白GmSqm及其编码基因与应用 |
| CA2977026A1 (en) * | 2015-03-11 | 2016-09-15 | E.I. Du Pont De Nemours And Company | Insecticidal combinations of pip-72 and methods of use |
| BR112017022000A2 (pt) | 2015-04-13 | 2018-07-03 | Bayer Cropscience Ag | derivados de n-cicloalquil-n-(biheterocicliletileno)-(tio)carboxamida. |
| CN104855410B (zh) * | 2015-05-20 | 2018-06-19 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| CN104920425B (zh) * | 2015-05-20 | 2018-06-19 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| AU2016297195A1 (en) * | 2015-07-23 | 2018-02-01 | Monsanto Technology Llc | Multi functional toxins |
| US10155960B2 (en) | 2015-08-27 | 2018-12-18 | Monsanto Technology Llc | Insect inhibitory proteins |
| US11841366B2 (en) | 2015-09-09 | 2023-12-12 | Syngenta Participations Ag | Compositions and methods for detection of wild type cry protein and a hybrid cry protein |
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| EP3960863A1 (en) * | 2016-05-04 | 2022-03-02 | Pioneer Hi-Bred International, Inc. | Insecticidal proteins and methods for their use |
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| US20190225974A1 (en) | 2016-09-23 | 2019-07-25 | BASF Agricultural Solutions Seed US LLC | Targeted genome optimization in plants |
| US20190261630A1 (en) | 2016-10-26 | 2019-08-29 | Bayer Cropscience Aktiengesellschaft | Use of pyraziflumid for controlling sclerotinia spp in seed treatment applications |
| AR109844A1 (es) * | 2016-10-27 | 2019-01-30 | Syngenta Participations Ag | Proteínas insecticidas |
| US11129906B1 (en) | 2016-12-07 | 2021-09-28 | David Gordon Bermudes | Chimeric protein toxins for expression by therapeutic bacteria |
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| ES2894278T3 (es) | 2017-06-13 | 2022-02-14 | Bayer Ag | 3-Fenilisoxazolin-5-carboxamidas de amidas de ácido tetrahidro- y dihidrofuranocarboxílico con efecto herbicida |
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| BR112020005311B1 (pt) | 2017-09-19 | 2024-02-27 | Bayer Aktiengesellschaft | Usos de isotianila, e método para controle do mal-do-panamá em plantas da família musaceae |
| JP2021505660A (ja) | 2017-11-30 | 2021-02-18 | ボラゲン，インコーポレーテッド | ベンゾキサボロール化合物およびその配合物 |
| CA3085324A1 (en) | 2017-12-15 | 2019-06-20 | Syngenta Participations Ag | Non-antibody ligands for detecting target proteins |
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| US11825850B2 (en) | 2020-12-21 | 2023-11-28 | Monsanto Technology Llc | Insect inhibitory proteins |
| UY39585A (es) | 2020-12-23 | 2022-07-29 | Monsanto Technology Llc | Proteínas que exhiben actividad inhibidora de insectos frente a plagas con importancia agrícola de plantas de cultivo y semillas |
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| EP4026833A1 (de) | 2021-01-12 | 2022-07-13 | Bayer Aktiengesellschaft | Herbizid wirksame 2-(het)arylmethylpyrimidine |
| AR126252A1 (es) | 2021-07-08 | 2023-10-04 | Bayer Ag | Amidas de ácido benzoico sustituidas |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5689052A (en) * | 1993-12-22 | 1997-11-18 | Monsanto Company | Synthetic DNA sequences having enhanced expression in monocotyledonous plants and method for preparation thereof |
| EP1103616A2 (en) * | 1989-02-24 | 2001-05-30 | Monsanto Company | Synthetic plant genes and method for preparation |
| WO2002014517A1 (en) * | 2000-08-11 | 2002-02-21 | Monsanto Technology Llc | BROAD-SPECTRUM δ-ENDOTOXINS |
| WO2003093484A1 (en) * | 2002-05-03 | 2003-11-13 | Bayer Bioscience N.V. | Insect resistant plants and methods for making the same |
| US20040093637A1 (en) * | 1996-11-20 | 2004-05-13 | Monsanto Technology Llc. | Hybrid Bacillus thuringiensis delta-endotoxins with novel broad-spectrum insecticidal activity |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5128130A (en) | 1988-01-22 | 1992-07-07 | Mycogen Corporation | Hybrid Bacillus thuringiensis gene, plasmid and transformed Pseudomonas fluorescens |
| US5055294A (en) | 1988-03-03 | 1991-10-08 | Mycogen Corporation | Chimeric bacillus thuringiensis crystal protein gene comprising hd-73 and berliner 1715 toxin genes, transformed and expressed in pseudomonas fluorescens |
| GB9318207D0 (en) | 1993-09-02 | 1993-10-20 | Sandoz Ltd | Improvements in or relating to organic compounds |
| US5545818A (en) | 1994-03-11 | 1996-08-13 | Calgene Inc. | Expression of Bacillus thuringiensis cry proteins in plant plastids |
| US5527883A (en) | 1994-05-06 | 1996-06-18 | Mycogen Corporation | Delta-endotoxin expression in pseudomonas fluorescens |
| US5593881A (en) | 1994-05-06 | 1997-01-14 | Mycogen Corporation | Bacillus thuringiensis delta-endotoxin |
| BR9713373A (pt) | 1996-11-20 | 2001-06-19 | Ecogen Inc | Delta-endotoxinas de amplo espectro |
| AU1241100A (en) | 1998-10-30 | 2000-05-22 | University Of Nebraska-Lincoln | Trans-species transfer of apoptotic genes and transgenic plants developed thereby |
| US6489542B1 (en) * | 1998-11-04 | 2002-12-03 | Monsanto Technology Llc | Methods for transforming plants to express Cry2Ab δ-endotoxins targeted to the plastids |
| KR100581163B1 (ko) | 2000-08-25 | 2006-05-22 | 신젠타 파티서페이션즈 아게 | 하이브리드 바실러스 튜린지엔시스 독소, 이를 암호화하는 핵산 및 이를 사용한 해충의 방제방법 |
| CN1134540C (zh) | 2001-06-22 | 2004-01-14 | 中国科学院微生物研究所 | 一种可使其产物分泌到细胞外的嵌合杀虫蛋白基因 |
-
2006
- 2006-08-30 KR KR1020087007734A patent/KR101156893B1/ko active IP Right Grant
- 2006-08-30 EP EP06813951A patent/EP1919935B1/en active Active
- 2006-08-30 CA CA2617803A patent/CA2617803C/en active Active
- 2006-08-30 NZ NZ566028A patent/NZ566028A/en unknown
- 2006-08-30 MX MX2008002802A patent/MX2008002802A/es active IP Right Grant
- 2006-08-30 AU AU2006284857A patent/AU2006284857B2/en active Active
- 2006-08-30 CN CN2006800316065A patent/CN101268094B/zh active Active
- 2006-08-30 CA CA2771677A patent/CA2771677A1/en not_active Abandoned
- 2006-08-30 ES ES06813951T patent/ES2400809T3/es active Active
- 2006-08-30 UA UAA200802497A patent/UA96421C2/ru unknown
- 2006-08-30 EA EA200800448A patent/EA015908B1/ru unknown
- 2006-08-30 PL PL06813951T patent/PL1919935T3/pl unknown
- 2006-08-30 WO PCT/US2006/033868 patent/WO2007027777A2/en active Application Filing
- 2006-08-30 AP AP2008004319A patent/AP2519A/xx active
- 2006-08-30 PT PT68139518T patent/PT1919935E/pt unknown
- 2006-08-30 US US12/064,875 patent/US8034997B2/en active Active
- 2006-08-30 BR BRPI0615657A patent/BRPI0615657B8/pt active IP Right Grant
- 2006-08-30 JP JP2008529233A patent/JP4975747B2/ja active Active
- 2006-08-30 CN CN201210257577.1A patent/CN102766652B/zh active Active
- 2006-08-31 AR ARP060103813A patent/AR055406A1/es active IP Right Grant
-
2008
- 2008-01-29 ZA ZA200800916A patent/ZA200800916B/xx unknown
- 2008-02-20 CR CR9747A patent/CR9747A/es unknown
- 2008-02-24 IL IL189718A patent/IL189718A/en active IP Right Grant
- 2008-02-27 EC EC2008008228A patent/ECSP088228A/es unknown
- 2008-02-28 HN HN2008000346A patent/HN2008000346A/es unknown
-
2011
- 2011-05-23 US US13/114,026 patent/US8344207B2/en active Active
-
2012
- 2012-11-27 US US13/686,320 patent/US20130095488A1/en not_active Abandoned
-
2017
- 2017-07-12 AR ARP170101939A patent/AR109260A2/es unknown
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1103616A2 (en) * | 1989-02-24 | 2001-05-30 | Monsanto Company | Synthetic plant genes and method for preparation |
| US5689052A (en) * | 1993-12-22 | 1997-11-18 | Monsanto Company | Synthetic DNA sequences having enhanced expression in monocotyledonous plants and method for preparation thereof |
| US6180774B1 (en) * | 1993-12-22 | 2001-01-30 | Monsato Company | Synthetic DNA sequences having enhanced expression in monocotyledonous plants and method for preparation thereof |
| US20040093637A1 (en) * | 1996-11-20 | 2004-05-13 | Monsanto Technology Llc. | Hybrid Bacillus thuringiensis delta-endotoxins with novel broad-spectrum insecticidal activity |
| WO2002014517A1 (en) * | 2000-08-11 | 2002-02-21 | Monsanto Technology Llc | BROAD-SPECTRUM δ-ENDOTOXINS |
| WO2003093484A1 (en) * | 2002-05-03 | 2003-11-13 | Bayer Bioscience N.V. | Insect resistant plants and methods for making the same |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112175093A (zh) * | 2014-10-16 | 2021-01-05 | 孟山都技术有限公司 | 对鳞翅目害虫具有毒性或抑制性的新型嵌合杀昆虫蛋白质 |
| CN104621171A (zh) * | 2015-01-30 | 2015-05-20 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| CN104621171B (zh) * | 2015-01-30 | 2018-10-30 | 北京大北农科技集团股份有限公司 | 杀虫蛋白的用途 |
| CN110214189A (zh) * | 2016-09-06 | 2019-09-06 | 农业生物群落股份有限公司 | 杀虫基因和使用方法 |
| CN110177464A (zh) * | 2016-12-20 | 2019-08-27 | 孟山都技术公司 | 新型昆虫抑制性蛋白 |
| CN110177464B (zh) * | 2016-12-20 | 2022-01-11 | 孟山都技术公司 | 新型昆虫抑制性蛋白 |
| CN114277037A (zh) * | 2016-12-20 | 2022-04-05 | 孟山都技术公司 | 新型昆虫抑制性蛋白 |
| CN110622998A (zh) * | 2019-10-14 | 2019-12-31 | 中国农业科学院植物保护研究所 | 一种蛋白在防治草地贪夜蛾和/或斜纹夜蛾中的应用 |
| CN110622998B (zh) * | 2019-10-14 | 2020-11-10 | 中国农业科学院植物保护研究所 | 一种蛋白在防治草地贪夜蛾和/或斜纹夜蛾中的应用 |
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