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Yongbin Qi Lei Chen Xiuling He Qingsheng Jin Xiaoming Zhang Zuhua He 《Pest management science》2013,69(1):135-141
BACKGROUND: Rice is the major food resource for nearly half of the global population; however, insect infestation could severely affect the production of this staple food. To improve rice insect resistance and reduce the levels of Bt toxin released into the environment, the Cry1Ab gene was conjugated to the rice rbcS promoter to express Bt toxin in specific tissues of transgenic plants. RESULTS: Eight marker‐free, T2 lines were separated from the T0 cotransformants. Using RT‐PCR, high levels of Cry1Ab expression were detected in the leaf but not in the seed. The Cry1Ab protein level ranged from 1.66 to 3.31 µg g?1 in the leaves of four transgenic lines, but was barely detectable in their seeds by ELISA. Bioassays showed that the mortality rate of silkworm larvae feeding on mulberry leaves dipped in transgenic rice flour and pollen was less than that of the positive control (KMD), and that their average weight was higher than that of KMD, suggesting that the Cry1Ab protein was not expressed in the seed and pollen. CONCLUSION: The transgene conferred a high level of resistance to insects and biosafety to the rice plants, which could be directly used in rice breeding. Copyright © 2012 Society of Chemical Industry 相似文献
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Bt蛋白在水稻不同遗传背景下的含量及对抗虫性的影响 总被引：1，自引：1，他引：0
为明确Bt基因在水稻杂交转育过程中抗虫作用的稳定性,以分别转入cry1C和cry2A基因的抗虫水稻品系T1C-19和T2A-1为供体亲本,与10个恢复系杂交并回交,筛选BC1F3代转基因纯合株系,用ELISA法测定水稻叶片和茎秆中Bt蛋白含量。结果表明,4个转cry1C株系和5个转cry2A株系叶片和茎秆的蛋白含量显著低于亲本,且降低了16.62%~62.03%;相同遗传背景下,叶片和茎秆中的Bt蛋白含量之间呈显著正相关;在室内转cry1C株系和转cry2A株系上二化螟幼虫的校正死亡率分别为62.50%~95.83%和37.50%~83.33%,其中在cry2A株系上的校正死亡率与茎秆中的Cry2A蛋白含量呈极显著正相关;在田间自然发虫条件下,纯合株系的单株卷叶数和枯心率最高仅为3.97和9.37%,受害程度与相应组织中Bt蛋白含量之间存在相关性。表明Bt蛋白含量会受到水稻遗传背景的影响,但杂交后代株系在田间仍能表现出很好的抗性。 相似文献
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Development and characterisation of transgenic rice expressing two Bacillus thuringiensis genes 总被引：1，自引：0，他引：1
BACKGROUND: Transgenic crops that produce insecticidal toxins from the bacterium Bacillus thuringiensis Berliner (Bt) were first commercialised in 1996. The risk that pests have the potential to evolve resistance to Bt toxins is one of the most serious challenges to this technology. Gene stacking, pyramiding two Bt genes into one variety, is considered to be an effective insect resistance management (IRM) strategy. In this study, insect‐resistant rice expressing two Bt genes was developed by sexual crossing, and then characterised. RESULTS: Homozygous rice lines of two pyramided Bt genes were obtained in the F3 generation. Quantification of Bt toxin showed that protein concentrations of Cry1Ab, Cry1Ac and Cry2A in the two‐gene lines were comparable with their single‐gene parents, while the expression of cry1C gene decreased after gene stacking. Four two‐gene lines showed higher activity to striped stem borer (Chilo suppressalis Walker) than parental lines in the laboratory bioassay. All pyramided lines and their hybrids exhibited excellent efficacy against stemborers and leaffolders in field evaluation, while most pyramided lines had no significant differences from original variety in yield under spraying of insecticide. CONCLUSION: These results demonstrate that the two‐gene lines have commercial potential and could serve as a valuable IRM strategy. Copyright © 2011 Society of Chemical Industry 相似文献
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为评价转Cry2Aa基因水稻T2A-1对二化螟盘绒茧蜂Cotesia chilonis的安全性,采用饲喂法和微注射法综合评价转Cry2Aa基因水稻T2A-1对二化螟盘绒茧蜂的影响。结果表明,寄生饲喂转Cry2Aa基因水稻T2A-1的二化螟Chilo suppressalis后,二化螟盘绒茧蜂体内未检出Cry2Aa蛋白,其每茧块茧数、茧长、雄成虫寿命和雌雄比分别为20.00个、2.58 mm、2.34 d和3.10,均显著小于对照,而卵及幼虫的总历期、蛹期、雌成虫寿命、茧质量和成虫质量均与对照差异不显著;寄生微注射Cry2Aa蛋白的二化螟后,除卵及幼虫的总历期和蛹期外,二化螟盘绒茧蜂其他生命表参数和体内过氧化物酶（peroxidase,POD）、超氧化物歧化酶（superoxide dismutase,SOD）和谷胱甘肽还原酶（glutathione reductase,GR）活力均与阳性对照之间差异显著;寄生微注射Cry2Aa蛋白的二化螟后,二化螟盘绒茧蜂所有生命表参数及体内POD、SOD和GR活力均与阴性对照之间无显著差异。表明饲喂法中转Cry2Aa基因水稻T2A-1对二化螟盘绒茧蜂产生的影响是由寄主介导效应引起的,而非Cry2Aa蛋白本身,因此转Cry2Aa基因水稻T2A-1对二化螟盘绒茧蜂安全。 相似文献
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采用人工饲料喂毒法和浸叶喂毒法测定了二化螟、大螟和稻纵卷叶螟初孵幼虫对4种Bt杀虫蛋白的敏感性。二化螟对Cry1Ab、Cry1Ac、Cry1C和Cry2A的LC50分别为1.739、2.6452、.794 ng/mg和15.965 ng/mg;大螟依次为2.455、10.046、11.803 ng/mg和35.359 ng/mg;稻纵卷叶螟依次为4.000、1.611、69.031μg/mL和12.708μg/mL。作者就不同水稻螟虫对不同Cry蛋白的敏感性差异及其在转基因抗虫作物中的应用作了讨论。 相似文献
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二化螟[Chilo suppressalis(Walker)]是一种钻蛀性害虫,具有食性杂、越冬场所多、转株为害等特点,其种群数量近年来呈回升趋势,危害逐年加重。二化螟对寄主存在明显的选择行为,其主要寄主之一水稻由于品种更替、耕作制度变更及气候变化等因素对二化螟种群发生规律产生了一些新的影响,给害虫防治工作带来了许多新问题。研究和了解二化螟的寄主选择规律及其机制,以及影响其种群动态的因素是有效调控二化螟种群的重要前提。本文就二化螟寄主选择行为与机制及其在水稻上种群消长影响因素等的研究概况进行综述报道,旨在更好地为二化螟的综合防治提供指导。 相似文献
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He YP Gao CF Chen WM Huang LQ Zhou WJ Liu XG Shen JL Zhu YC 《Pest management science》2008,64(3):308-315
BACKGROUND: Chemical control is a major strategy for suppressing the rice stem borer, Chilo suppressalis (Walker). Owing to their high toxicity and increasing resistance development in the target insect, many insecticides will be phased out entirely in 2007 in China. Alternatives with relatively low toxicity are urgently needed to replace traditional chemicals for rice stem borer control. In this study, the authors examined four field populations of C. suppressalis for their toxicological responses to more than 20 insecticides, including a few low-toxicity organophosphates and many novel pesticides. Interpopulation resistance levels to 12 conventional insecticides were also compared. RESULTS: Based on LD(50) values, the rice stem borer was most sensitive to avermectins and fipronil (LD(50) < 1 ng larva(-1)). The stem borers exhibited the least sensitivity to endosulfan (LD(50) > 100 ng larva(-1)) and monosultap (LD(50) > 1000 ng larva(-1)). Insect growth regulators and chitin synthase inhibitors showed great efficacy against C. suppressalis, especially against populations that had developed resistance to conventional insecticides. Four field populations showed variable tolerance levels to many insecticides. LYG05 was the most susceptible population, only with a low level of resistance to monosultap (RR = 6.6). NC05 and GL05 populations exhibited intermediate tolerance levels with RR values up to 20.4 and 52.8 respectively. RA05 was the most resistant population to many insecticides, with resistance ratios up to 76.2. CONCLUSION: The results from this study provide valuable information for selection and adoption of new alternative insecticides and for resistance management of the rice stem borer. 相似文献
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研究了cryLA(b)基因及其介导的抗虫性在转基因水稻自交或回交后代中的遗传规律和遗传稳定性。结果表明：cryLA(b)基因及其连锁的bar基因作为一个显性遗传位点遵循孟德尔遗传分离规律；在T1代群体中遵循3：1的分离规律，T2代稳定纯合；在BC11～BC4代群体中遵循1：1的分离规律，BC群体自交的BCF9群体中遵循3：1分离规律。cryLA(b)基因及其介导抗性已稳定地遗传至T7代和BC4代。室内及田间抗虫性鉴定显示，不同世代的转基因植株均保持稳定的高度抗虫性。 相似文献
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为明确转cry1Ab+cry1C双价抗虫水稻对二化螟的抗性及其杀虫蛋白的时空表达,本研究采用离体水稻组织生测法系统评价了cry1Ab、cry1C、正交cry1Ab+cry1C和反交cry1C+cry1Ab 4种抗虫水稻品系对二化螟的杀虫效果,并用ELISA方法测定了Cry1Ab、Cry1C蛋白在各个抗虫水稻品系中的表达情况。结果显示,二化螟在不同生育期的4种转基因抗虫水稻上取食的叶面积显著低于非转基因对照亲本明恢63。4个转基因抗虫水稻品系在生长前期（苗期、分蘖和拔节期）对二化螟表现极高的杀虫效果,生长后期（孕穗期和成熟期）杀虫效果有所下降。两双价抗虫水稻杀虫效果最好,其次为转cry1Ab和cry1C水稻。Bt蛋白表达水平随着抗虫水稻生育期的变化而变化,且差异显著;Cry1Ab的蛋白表达量在水稻整个生长期均显著高于Cry1C。Cry1Ab蛋白在单价抗虫水稻叶片和茎杆中均表现出随生育期先升高后下降的趋势,但在双价抗虫水稻中表现出随生育期逐渐下降的趋势。Cry1C蛋白在单、双价抗虫水稻的叶片组织中均表现出逐渐升高的趋势。与单价抗虫水稻相比,双价抗虫水稻中的Cry1Ab、Cry1C的蛋白表达水平并没有表现出显著的降低。因此,双价抗虫水稻不仅能高效防治害虫,还能延缓害虫抗性,在生产上表现出良好的应用前景。该研究结果可望为转Bt基因抗虫作物的环境安全评价提供科学数据和理论依据。 相似文献
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BACKGROUND: Extensive adoption of transgenic Bt corn in recent years for stalk borer control has increased risk of resistance evolution in the target pest populations. A Bt‐resistant strain of the sugarcane borer, Diatraea saccharalis, was approximately 100‐fold more tolerant to Cry1Ab toxin than the susceptible counterpart. To gain a better understanding of the molecular mechanisms of Bt resistance, the Cry1Ab‐susceptible (Cry1Ab‐SS) and Cry1Ab‐resistant (Cry1Ab‐RR) strains of D. saccharalis were subjected to a microarray analysis. RESULTS: Results showed that the expression levels of many genes were significantly different between the Cry1Ab‐RR and Cry1Ab‐SS strains. Microarray analysis of 7145 cDNAs revealed 384 differentially expressed genes. A total of 273 genes were significantly upregulated 2–51.6‐fold, and 111 genes were significantly downregulated 2–22.6‐fold in the Cry1Ab‐RR strain. The upregulation of three potential resistance‐related genes, coding for a glutathione S‐transferase (GST), a chymotrypsin‐like protease (CHY) and a lipase (LP), was confirmed using real‐time PCR, indicating a reproducibility of the microarray data. Ontology analysis revealed that more than twice the number of metabolic‐related genes were upregulated compared with downregulated genes with the same biological function. Up to 35.2% of the upregulated genes in the resistant strain were associated with catalytic activity, while only 9.5% of the downregulated genes were related to the same catalytic molecular function. CONCLUSION: The large portion of metabolic‐ or catalytic‐related genes with significant upregulations indicated a potential large increase in metabolic or catalytic activities in the Cry1Ab‐RR strain. This cDNA microarray gene expression data could be used to characterize and identify new genes that may be associated with Bt resistance in D. saccharalis. Copyright © 2012 Society of Chemical Industry 相似文献
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水稻害虫上三种病原镰刀菌的研究 总被引：1，自引：0，他引：1
1979～1982年间,作者在对湖南水稻主要害虫病原微生物资源调查中,发现鳞翅目、同翅目和双翅目中的许多种害虫被镰刀菌属的串珠镰刀菌(Fusari-um moniliforme)、燕麦镰刀菌(F.avenaceum)及本色镰刀菌(F.Concolor)所寄生。尤其是在每年3～4月间对越冬螟虫尸体上的病原物分离中,镰刀菌占90%以上。用这些菌种对二化螟、大螟越冬幼虫进行生物测验,其寄生死亡率在50～100%之间;对褐稻虱和黑尾叶蝉的寄生率为50～80%。三种镰刀菌发酵物的浸提液对害虫都有一定的毒力,其中以燕麦镰刀菌的最强。用该菌在玉米粒上培养一年的发酵物,以湿重5克加水150毫升的浸提液饲养中华大蟾蜍的蝌蚪,3～4小时内全部死亡。用此液处理稻苗饲养二化螟幼虫,死亡率为64%左右;用处理的甘蓝叶饲喂菜青虫,死亡率为70%左右。将燕麦镰刀菌发酵物浸提液在-16℃下冷冻5～6天和在100℃时煮沸5～10分钟仍不丧失其毒力。 相似文献
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水稻螟虫是我国水稻生产过程中的重要害虫种类之一, 其为害范围广, 所造成的经济损失巨大, 且在我国主要稻区呈现逐年加重的发生趋势。本文重点以二化螟为例, 概述了近年来水稻螟虫发生与为害现状、抗药性演化、抗螟水稻种质资源挖掘与品种创制、水稻螟虫生理生化与毒理学研究及新型杀螟药剂研发、水稻螟虫寄生蜂资源及其利用、水稻螟虫生态调控技术等方面的主要研究成果, 以期为深入了解我国水稻螟虫发生为害及其治理研究现状提供依据, 并为水稻螟虫的可持续控制与水稻绿色生产的实践提供帮助。 相似文献
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Field‐evolved resistance to Bt maize in sugarcane borer (Diatraea saccharalis) in Argentina 
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下载免费PDF全文 Damián A Grimi Betiana Parody María Laura Ramos Marcos Machado Federico Ocampo Alan Willse Samuel Martinelli Graham Head 《Pest management science》2018,74(4):905-913
BACKGROUND
Maize technologies expressing Bacillus thuringiensis (Bt) insecticidal proteins are widely used in Argentina to control sugarcane borer (Diatraea saccharalis Fabricius). Unexpected D. saccharalis damage was observed to Bt maize events TC1507 (expressing Cry1F) and MON 89034 × MON 88017 (expressing Cry1A.105 and Cry2Ab2) in an isolated area of San Luis Province. Diatraea saccharalis larvae were sampled from MON 89034 × MON 88017 fields in the area to generate a resistant strain (RR), which was subsequently characterized in plant and diet bioassays.RESULTS
Survivorship of the RR strain was high on TC1507 leaf tissue, intermediate on MON 89034 × MON 88017, and low on MON 810 (expressing Cry1Ab). The RR strain had high resistance to Cry1A.105 (186.74‐fold) and no resistance to Cry2Ab2 in diet bioassays. These results indicate resistance to Cry1F and Cry1A.105 (and likely cross‐resistance between them) but not to Cry1Ab or Cry2Ab2. Resistance to MON 89034 × MON 88017 was functionally recessive. Reviews of grower records suggest that resistance initially evolved to Cry1F, conferring cross‐resistance to Cry1A.105, with low refuge compliance as the primary cause. A mitigation plan was implemented in San Luis that included technology rotation, field monitoring, and grower education on best management practices (BMPs) including refuges.CONCLUSION
In the affected area, the resistance to Cry1F and Cry1A.105 is being managed effectively through use of MON 89034 × MON 88017 and MON 810 in combination with BMPs, and no spread of resistance to other regions has been observed. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. 相似文献15.
Characterization of field‐evolved resistance to Bacillus thuringiensis‐derived Cry1F δ‐endotoxin in Spodoptera frugiperda populations from Argentina 下载免费PDF全文
下载免费PDF全文 Desmi I Chandrasena Ana M Signorini Gustavo Abratti Nicholas P Storer Magdalena L Olaciregui Analiza P Alves Clinton D Pilcher 《Pest management science》2018,74(3):746-754
BACKGROUND
Transgenic maize (Zea mays L.) event TC1507 (Herculex® I insect protection), expressing Cry1F δ‐endotoxin derived from Bacillus thuringiensis var. aizawai, was commercialized in 2003 in the Americas. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) susceptibility to Cry1F was monitored annually across several regions in Argentina using diagnostic concentration bioassays. Reduced performance of TC1507 maize against S. frugiperda was reported in 2013. A resistant population was established in the laboratory and the dominance of Cry1F resistance was characterized.RESULTS
During 2012–2015, high‐survivorship of several populations was observed in the resistance monitoring program. Reciprocal crosses of a Cry1F‐resistant population with a Cry1F‐susceptible population were evaluated to calculate effective dominance (DML) based on mortality levels observed at 100 µg/ml Cry1F. Two additional dominance levels (DLC and DEC) were calculated using lethal (LC50) or effective concentration (EC50) derived from concentration–response bioassays. Estimates indicated that Cry1F resistance in S. frugiperda in Argentina was either highly recessive (DML = 0.005) or incompletely recessive (DLC < 0.26 and DEC < 0.19).CONCLUSION
This study is the first documented confirmation and characterization of S. frugiperda Cry1F field‐evolved resistance in Argentina. The resistance to Cry1F in S. frugiperda populations collected in Argentina, is autosomal and incompletely recessive similar to the resistance reported in Brazil. © 2017 The Authors. Pest Management Science published by John Wiley © Sons Ltd on behalf of Society of Chemical Industry. 相似文献16.
玉米是全球播种面积最大的粮食作物,事关世界粮食安全。鳞翅目和鞘翅目害虫是影响玉米产量和品质的重要因素,长期以来多数国家采取以喷施化学农药为主的防控策略,但存在较高的使用成本和环境风险等问题。作为新一代的害虫防控技术,转基因抗虫玉米于1996年开始在美国商业化种植,并迅速推广到巴西等主要玉米生产国家,成为防控草地贪夜蛾Spodoptera frugiperda和欧洲玉米螟Ostrinia nubilalis等重大害虫的核心技术。该文综述了全球转基因抗虫玉米商业化的历史和防控害虫的作用,分析了美国对欧洲玉米螟和草地贪夜蛾等靶标害虫抗性治理的成功经验以及巴西等南美国家草地贪夜蛾对多种转基因抗虫玉米产生抗性的成因与教训。基于中国转基因抗虫玉米转化事件的研发现状、玉米生产模式、玉米害虫的区域发生特点和迁飞生物学等特性,提出在南方和西南山地丘陵玉米区种植包含Vip3A的多基因叠加抗虫玉米从源头防控草地贪夜蛾,在黄淮海夏玉米区种植包含Cry2A的多基因叠加抗虫玉米从源头防控棉铃虫Helicoverpa armigera,在北方春玉米区种植包含Cry1A的多基因叠加抗虫玉米从源头防控亚洲玉米螟Ostr... 相似文献
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转cry1Ac玉米对亚洲玉米螟的抗性评价 总被引：1，自引：1，他引：0
目标性状的有效性评价是转Bt基因抗虫玉米研发和安全性评价研究的重要内容之一。采用酶联免疫吸附剂测定（ELISA）、室内生测和田间人工接虫鉴定研究了转cry1Ac玉米纯合群体BT-X和分离群体BT-38、BT-181、BT-43、BT-105等转化事件Cry1Ac蛋白的表达量以及对亚洲玉米螟Ostrinia furnacalis（Guenée）的室内杀虫和田间抗虫效果。转cry1Ac玉米鲜组织中Cry1Ac蛋白含量在44.07～438.00 ng/g之间,不同转化事件及不同组织之间差异显著,其中BT-X鲜花丝中Bt蛋白表达量显著低于未展开心叶。室内生测试验中除BT-43与对照没有显著差异外,取食其它转基因玉米心叶的亚洲玉米螟初孵幼虫6或7天的存活率为0～15.3%,显著低于非转基因对照;取食夏播BT-X鲜花丝6天的幼虫存活率为62.5%,显著低于Bt11及非转基因对照。田间心叶期接虫鉴定显示BT-X高抗亚洲玉米螟。说明转cry1Ac玉米BT-X、BT-181、BT-38和BT-105对亚洲玉米螟有很高的杀虫作用,BT-X田间抗虫效果良好。 相似文献
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BACKGROUND: Studies of hybrid fitness, of which agronomic performance may be an indicator, can help in evaluating the potential for introgression of a transgene from a transgenic crop to wild relatives. The objective of this study was to assess the agronomic performance of reciprocal hybrids between two transgenic glufosinate‐resistant rice lines, Y0003 and 99‐t, and two weedy rice accessions, WR1 and WR2, in the greenhouse. RESULTS: F1 hybrids displayed heterosis in height, flag leaf area and number of spikelets per panicle. The agronomic performance of F1 between WR1 and Y0003 was not affected by crossing direction. The tiller and panicle numbers of F1 individuals were higher than their F2 counterparts. However, these traits did not change significantly from the F2 to the F3 generation or in hybrids with weedy rice as maternal or paternal plants. For all hybrids, the in vitro germination rates of fresh pollen were similar and significantly lower than those of their parents, seed sets were similar to or of lower value than those of weedy rice parents and seed shattering characteristics were partially suppressed, but the survival of hybrids over winter in the field was similar to that of weedy rice parents. All F1, F2 and F3 hybrids had similar composite agronomic performance to weedy rice parents. CONCLUSION: There was no significant decrease in the composite agronomic performance of any of the hybrids compared with weedy rice. This implies that gene flow from transgenic cultivated rice to weedy rice could occur under natural conditions. Copyright © 2011 Society of Chemical Industry 相似文献
19.
为获得新型抗虫转基因玉米,将通过群体筛选获得的具有抗虫性的转cry2Ah-vp基因玉米VP1-5采用PCR、Southern blot、实时荧光定量PCR(qPCR)、酶联免疫吸附测定(ELISA)等方法进行阳性植株鉴定、拷贝数分析、转录水平和翻译水平分析,同时通过室内和田间生物活性测定鉴定转基因玉米VP1-5对东方黏虫Mythimna separata和亚洲玉米螟Ostrinia furnacalis的抗性。结果表明,在转基因玉米VP1-5中cry2Ah-vp基因已整合到玉米基因组,以单拷贝的形式插入;cry2Ah-vp基因在转基因玉米VP1-5不同部位组织中均可以正常转录,在灌浆期叶片中的mRNA表达量最高,相对表达量为32.67,在灌浆期穗轴中的mRNA表达量最低,相对表达量为3.74;Cry2Ah-vp蛋白在转基因玉米VP1-5的6叶期各组织中表达量均较高,其中在叶片中的表达量达到2 155.18 ng/g FW,在抽雄期花丝中的表达量最高,达到2 165.86 ng/g FW;且转基因玉米VP1-5对东方黏虫有很高的杀虫活性,接虫3 d后幼虫死亡率达到100.00%;对亚洲玉米螟幼虫也有明显的生长抑制作用。表明转基因玉米VP1-5可作为玉米抗虫育种和害虫防治的种质资源。 相似文献
20.
玉米自交系对亚洲玉米螟抗性评价初报 总被引：1，自引：0，他引：1
在相同田间条件下,用接种方法评价了一些玉米自交系对第一代亚洲玉米螟[Ostrinia furnacalis(Guenee)]的抗性。经鉴定抗螟的有L105、75—242、B75、W59E、404、黄早4、A662、C131A和4004等。它们与感虫自交系比分别减少螟虫幼虫57.8、64.4、55.6、37.8、50.0、71.4、64.3、0和20%,茎蛀孔减少68.3、61.9、55.6、38.1、63.8、57.4、72.3、66.0和47.6%,隧道长度分别减少70.1、77.5、53.2、34.6、71.3、76.6、40.7、77.0和32.9%。茎折、穗和穗柄被害比感虫自交系也明显减轻。 相似文献