Whole-genome sequencing to detect mutations associated with resistance to insecticides and Bt proteins in Spodoptera frugiperda

The fall armyworm (FAW), Spodoptera frugiperda, is a major pest native to the Americas that has recently invaded the Old World. Point mutations in the target-site proteins acetylcholinesterase-1 (ace-1), voltage-gated sodium channel (VGSC) and ryanodine receptor (RyR) have been identified in S. frugiperda as major resistance mechanisms to organophosphate, pyrethroid and diamide insecticides respectively. Mutations in the adenosine triphosphate-binding cassette transporter C2 gene (ABCC2) have also been identified to confer resistance to Cry1F protein. In this study, we applied a whole-genome sequencing (WGS) approach to identify point mutations in the target-site genes in 150 FAW individuals collected from China, Malawi, Uganda and Brazil. This approach revealed three amino acid substitutions (A201S, G227A and F290V) of S. frugiperda ace-1, which are known to be associated with organophosphate resistance. The Brazilian population had all three ace-1 point mutations and the 227A allele (mean frequency = 0.54) was the most common. Populations from China, Malawi and Uganda harbored two of the three ace-1 point mutations (A201S and F290V) with the 290V allele (0.47–0.58) as the dominant allele. Point mutations in VGSC (T929I, L932F and L1014F) and RyR (I4790M and G4946E) were not detected in any of the 150 individuals. A novel 12-bp insertion mutation in exon 15 of the ABCC2 gene was identified in some of the Brazilian individuals but absent in the invasive populations. Our results not only demonstrate robustness of the WGS-based genomic approach for detection of resistance mutations, but also provide insights for improvement of resistance management tactics in S. frugiperda.     

转基因欧洲黑杨中Bt基因表达特性的研究

利用PCR检测了转基因欧洲黑杨当代和杂交后代群体 ,外源Bt基因在 7年生的转基因欧洲黑杨 1 4个无性系中稳定存在 ,Bt基因在杂交后代中分离比例为 1∶1。转基因欧洲黑杨作母本进行杂交时有落花、落果现象 ,可能是基因改造影响了它们的某些生理功能。     

加工产品中转基因玉米Bt11成分实时荧光PCR定量(性)检测

实验在玉米自身基因和外源基因的边界序列之间设计了具有品种和品系特异性的引物和探针 ,并以实时荧光PCR技术 ,建立了加工产品中转基因玉米Bt1 1成分品系鉴定检测和定量检测的方法。实验对加热条件和时间对检测转基因成分的影响作了探讨 ,并检测了部分市售食品和饲料。检测结果发现 ,加热时间温度越高、时间越长 ,对转基因成分定量检测的影响越大 ;在所检测的样品中可以检测出转基因玉米Bt1 1成分 ,有些样品还同时检出其他转基因成分。本研究实验建立的方法 ,可以用于加工产品中转基因成分的定量检测 ,也可以用于定性检测 ,或作为常规PCR定性检测后的确证实验方法。     

A decade of <Emphasis Type="Italic">Bt</Emphasis> cotton in Chinese fields: Assessing the direct effects and indirect externalities of Bt cotton adoption in China

The objective of this study is to examine whether or not the gains from reduced spraying for bollworms are being sustained more than one decade after the initial adoption in 2007. Based on farm-level data collected by the authors in 1999–2007 in 16 villages from 4 provinces, this study shows that insecticides applied for controlling bollworms have declined. This analysis supports Chinese policy makers’ decision to not require refuges of non-Bt cotton fields. It also suggests that past studies may have underestimated the benefits from adopting Bt technology.     

棉铃虫和甜菜夜蛾中肠丝氨酸蛋白酶活性测定以及活化、降解Cry1Ac毒素分析

昆虫中肠对Bt原毒素活化与对活化毒素降解的变化被认为是害虫对Bt产生的机制之一,研究比较棉铃虫Helicoverpa armigera(Hübner)与甜菜夜蛾Spodoptera exigua(Hübner)的中肠液、BBMV蛋白酶的活性,通过SDS-PAGE分析2种昆虫对原毒素的活化速度与对活化毒素的降解速度。2种昆虫的中肠液蛋白酶活性均显著高于BBMV蛋白酶活性,中肠液与BBMV均能迅速活化原毒素并继续降解活化后的毒素,与中肠液相比,BBMV对原毒素的活化与对活化毒素的降解均慢于中肠液,甜菜夜蛾对毒素的活化与降解又慢于棉铃虫。另外,还测定抑制剂对中肠液蛋白酶活性的抑制作用,结果表明,各抑制剂对棉铃虫和甜菜夜蛾相应酶活性的抑制表现出相同的趋势,TLCK对丝氨酶蛋白酶具较好的抑制作用,而PMSF对胰蛋白酶的抑制作用次之,TPCK对胰凝乳蛋白酶的抑制作用较弱。     

植物的抗螨机理

植食性螨类分布广泛,危害多种寄主植物.本文从3个方面介绍了植物的抗螨机理:植物形态如叶型、叶色主要影响螨类的取食产卵位点和营养物质的获取;植物组织结构如叶厚、蜡质、茸毛、气孔等决定螨能否顺利利用口针取食到植物的汁液,而虫菌穴则作用于捕食螨和植食性螨的相互关系,从而达到保护植物的目的;植物理化性质如叶鲜重、生长速度、生育...     

田间转Bt基因水稻上稻飞虱卵量、孵化率及天敌作用

为了评估转基因水稻释放后对非靶标害虫及其天敌的影响, 本研究在浙江大学实验农场和中国水稻研究所实验农场2008和2009年连续两年监测了转Bt基因水稻对田间稻飞虱着卵量、卵孵化率、卵被捕食率、卵被寄生率和卵死亡率的影响。结果表明: 两年2个调查地点3个水稻品系（克螟稻1、克螟稻2和秀水11）上的稻飞虱平均每分蘖着卵量、卵孵化率、卵被捕食率和卵被寄生率的趋势基本一致, 而且田间3个水稻品系上稻飞虱卵量的高峰期均持续2周左右; 但是两年2个调查地点3个水稻品系上稻飞虱卵死亡率则没有一致的趋势。此外, 总体上3个水稻品系上稻飞虱平均每分蘖着卵量、卵孵化率、卵被捕食率、卵被寄生率和卵死亡率差异均不显著（P>0.05）。因此, 稻田中转Bt基因水稻对稻飞虱卵没有显著的影响。     

Zero effect of Bt rice on expression of genes coding for digestion,detoxification and immune responses and developmental performances of Brown Planthopper Nilaparvata lugens (Stål)

Transgenic Cry1Ac, Cry2Aa and Cry1Ca (Bt toxins) rice lines are well developed to manage lepidopteron pests in China. The impact of transgenic Bt rice on the non-target Brown Planthopper (BPH) has become an essential part of environmental risk assessment, however, scanty evidence is found addressing on developmental and molecular responses of BPH to the ingestion of Bt protein from transgenic rice. The focus of the current study is to examine the developmental characteristics and the expression profiles of gene in relation to digestion, detoxification and immune responses were examined. Our study strongly revealed that the tested Bt rice strains have no unfavorable effect on fecundity, survival and growth of BPH. Furthermore, each of the tested genes did not exhibit distinct expression pattern responding to non Bt parental cultivar, thus, it could be concluded that Bt rice have no detrimental effects on the physiological processes of digestion, detoxification and immune responses of BPH.     

转基因棉种植对土壤水解酶活性的影响

转Bt基因棉和转Bt＋cpTI基因棉种植面积不断扩大,它们种植后杀虫晶体蛋白在土壤中的残留特性及对土壤水解酶活性的影响是环境风险评价的重要组成部分,本文采用盆栽实验的方法对此进行了初步研究。结果表明,转基因棉出苗后生长到30天时可向土壤中释放Bt杀虫晶体蛋白,而双抗棉种植时CpTI杀虫晶体蛋白的释放量与品种有关;转基因棉出苗后30天时,与等价基因系非转基因棉（各对照）相比,转Bt基因棉（“中30”）和双抗棉A（转Bt＋CpTI棉“中41”）的种植并未使脲酶、蛋白酶和磷酸单脂酶活性发生显著变化,而双抗棉B（转Bt＋CpTI棉“双抗321”）的种植使土壤磷酸单脂酶活性显著下降。从杀虫晶体蛋白的释放和对酶活性的影响来看,双抗棉A的种植对土壤的生物活性扰动更小。     

Climatic limits of pink bollworm in Arizona and California: effects of climate warming

The distribution and abundance of pink bollworm (Pectinophora gossypiella Saunders (PBW)) in cotton in Arizona and California was examined using a validated weather-driven, physiologically based demographic model of cotton and PBW integrated into a geographic information system (GIS). Survival of diapause larvae during winter as affected by low temperatures is a key factor determining the range of PBW. Winter survival was estimated using data from Gutierrez et al. (Can. Entomol. 109 (1977) 1457) and Venette et al. (Environ. Entomol. 29 (5) (2000) 1018). The model was run continuously over the period 1 January 1995 to 31 December 2003 using observed weather data from 121 locations. Three output variables were mapped as measures of PBW persistence: over-winter survival of diapause PBW larvae, cumulative daily PBW larval densities over the season, and the number of diapause larvae produced during the season. The distribution of pink bollworm is predicted to be restricted to the relatively frost-free cotton growing areas of Arizona and Southern California where it currently reaches pest status. The model predicts that extension of PBW's range into the Central Valley of California is unlikely. The analysis questions the efficacy of an ongoing area-wide effort to prevent the establishment of PBW in the Central Valley of California. Four global warming scenarios were examined to estimate the effects on the potential geographic range of PBW. Average observed daily temperatures were increased 1.0, 1.5, 2.0 or 2.5 °C, respectively, in the four scenarios. Scenarios with average increases of 1.5–2.5 °C predicted that the range of PBW would expand into the Central Valley of California and the severity of the pest would greatly increase in areas of current infestation.