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1.
Cry1Ab杀虫蛋白在水稻-褐飞虱-拟水狼蛛食物链中转移与富集   总被引:17,自引:0,他引:17  
采用ELISA方法检测了2个转cry1Ab基因水稻(Bt水稻)品系KMD1和KMD2不同生育期叶鞘内Cry1Ab杀虫蛋白的含量及其通过褐飞虱和拟水狼蛛的转移和富集情况。结果表明,这2个品系中抽穗期和黄熟期叶鞘内Cry1Ab的含量均显著低于苗期、分蘖期和孕穗期,KMD1和KMD2中Cry1Ab杀虫蛋白可以通过食物链转移到Bt水稻非靶标害虫褐飞虱及其天敌拟水狼蛛体内。褐飞虱在KMD1或KMD2上取食2 d后,体内均含有Cry1Ab杀虫蛋白,但连续取食2、4、6、8和10 d后,其体内含量并未因取食时间的延长而呈现明显增加的趋势。当拟水狼蛛捕食以KMD1或KMD2为食的褐飞虱时,在捕食2、4、6、8和10 d后,其体内均可检测到Cry1Ab杀虫蛋白,其含量并未随捕食时间的延长而明显上升,但均显著高于相应时间褐飞虱体内的含量。可见,该蛋白可通过水稻转移至褐飞虱,再转移至拟水狼蛛,并存在明显的富集现象,而这种富集并不随蜘蛛捕食时间的延长而加强。拟水狼蛛捕食以KMD1或KMD2为食的褐飞虱时,其捕食量未受到显著影响,其中肠酶粗提物对Cry1Ab杀虫蛋白具有明显的降解作用。  相似文献   

2.
王世贵  叶恭银  胡萃 《昆虫学报》2006,49(2):200-205
研究了转cry1Ab基因水稻“克螟稻1号”对二化螟Chilo suppressalis幼虫细胞免疫系统的影响。结果表明,转cry1Ab基因水稻对二化螟幼虫的血细胞影响明显,取食转cry1Ab基因水稻后,二化螟幼虫各类血细胞都明显低于取食非转基因水稻“秀水11”的对照组(原血细胞和囊血细胞在取食初期例外),随取食时间延长,各类血细胞数量及血细胞总数均呈递减的趋势。从各类血细胞所占血细胞总数的百分比来看,原血细胞在取食36 h后锐减,而浆血细胞和粒血细胞则比例增加,其余珠血细胞、囊血细胞的变化不明显。另外,血细胞还出现空泡化、肿胀等病态变化,致使血细胞快速破裂。由此推测转cry1Ab基因水稻自身表达的毒蛋白能严重干扰靶标昆虫二化螟幼虫的细胞免疫系统。  相似文献   

3.
为明确转基因水稻表达的Bt蛋白沿捕食性食物链的传递及其对生态系统中较高营养级生物的影响,在实验条件下(28±0.5 ℃,R.H=80%,L∶D=12h∶12h)以田间优势自然天敌拟环纹豹蛛为对象,以取食转Cry1Ab/Cry1Ac基因水稻汕优63(简称Bt水稻)叶片24 h的稻纵卷叶螟幼虫为猎物饲养拟环纹豹蛛,并测定了...  相似文献   

4.
Cry1Ab蛋白对稻纵卷叶螟幼虫体内三种保护酶活性的影响   总被引:1,自引:0,他引:1  
室内采用酶活力测定方法研究转B t基因水稻表达的Cry1Ab蛋白对稻纵卷叶螟幼虫体内3种保护酶(AChE、SOD和CAT)活性的影响。结果表明,用转B t基因水稻处理稻纵卷叶螟幼虫,饲喂4 h、36 h后,其体内AChE活力分别比对照增加了52.09%、128.51%,并且都极显著(P<0.01)高于对照组;取食转基因水稻叶片4 h后,幼虫体内SOD酶活性比对照提高了91.5%,与对照有显著差异;36 h后活性达到最大值,但与对照差异不显著。取食转基因水稻叶片24 h后,幼虫体内CAT酶活性达到最大值且高于对照,但与对照差异不显著;48 h后酶活性受到显著抑制,与对照差异显著。同时,测定了幼虫体内及其粪便中Cry1Ab蛋白含量的变化,结果表明,取食转基因水稻叶片12 h后,进入体内的Cry1Ab蛋白随粪便排出,幼虫体内的Cry1Ab蛋白含量一直低于粪便中的含量,且在24 h时两者差异达到最大。由于Cry1Ab蛋白在幼虫体内的积累,扰乱了稻纵卷叶螟幼虫体内AChE、SOD和C AT保护酶的动态平衡,使虫体内自由基的清除遇到障碍,从而对其产生毒害作用。  相似文献   

5.
转Bt基因水稻对二化螟绒茧蜂生物学特性的影响   总被引:10,自引:4,他引:6       下载免费PDF全文
用转Bt基因 (cry1Ab) 水稻KMD1饲喂3龄、4龄和5龄二化螟Chilo suppressalis一定时间后, 作为二化螟绒茧蜂Apanteles chilonis的寄主,研究了转基因水稻经寄主对绒茧蜂生物学特性的影响。结果发现:KMD1处理后,三个龄期幼虫的寄生率都显著下降,其中4龄和5龄达极显著水平;3龄和4龄上的结茧率显著低于对照;蜂蛹历期均短于对照,但仅3龄差异显著;从5龄幼虫所羽化的雄蜂寿命显著短于对照;蜂茧长显著短于对照;而对卵+幼虫期、茧块茧数、蜂羽化率及性比均无显著影响。  相似文献   

6.
在实验室研究了转sck+cry1Ac基因水稻(MSB)对二化螟Chilo suppressalis (Walker)生长、存活以及经寄主对二化螟绒茧蜂Apanteles chilonis (Munakata)生物学特性的影响。连续取食转sck+cry1Ac基因水稻的二化螟, 体重下降、死亡率上升,从第2天开始,其体重显著低于取食明恢86的对照组;从第6天开始,死亡率极显著高于对照组。二化螟取食MSB 36 h后移至对照水稻上继续取食3、6、9、12天后,死亡率与对照差异都不显著;但体重均低于对照,其中第3天的体重差异达显著水平。二化螟绒茧蜂分别以取食MSB一定时间的3、4、5龄二化螟幼虫为寄主时,寄生率均低于以对照组,其中对4龄幼虫的寄生率差异显著;结茧率与对照差异均不显著;寄生在取食MSB的5龄二化螟幼虫体内的蜂、蛹期显著长于对照,而所结茧的茧长显著短于对照;但卵 幼虫历期、每茧块茧数、羽化率、雌性率、成蜂寿命和前翅长与对照无显著差异。结果表明转sck+cry1Ac基因水稻不仅对二化螟生长和存活有显著影响, 而且可经寄主二化螟影响到二化螟绒茧蜂的一些生物学特性。  相似文献   

7.
张巍  张志罡  付秀芹  刘立军  颜亨梅 《昆虫学报》2008,51(10):1022-1027
用转Cry1Ab/Cry1Ac基因水稻的叶片饲喂稻纵卷叶螟Cnaphalocrocis medinalis (Guenée)幼虫,采用酶活力测定方法研究了转基因水稻对稻纵卷叶螟幼虫体内3种保护酶(SOD,CAT和POD)活性的影响。结果表明:取食转基因水稻叶片4 h后,幼虫体内SOD酶活性比对照提高了91.5%,与对照有显著差异;36 h后活性达到最大值,但与对照差异不显著。取食转基因水稻叶片24 h后,幼虫体内CAT酶活性达到最大值且高于对照,但与对照差异不显著;48 h后酶活性受到显著抑制,与对照差异显著。取食转基因水稻叶片12 h后,幼虫体内POD酶活性达到最大值,与对照差异不显著;48 h后酶活性逐渐下降达到最小值,比对照下降68.05%。实验同时测定了幼虫体内及其粪便中Bt毒蛋白含量的变化,结果表明取食转基因水稻叶片12 h后,随着大量取食进入体内的毒蛋白随粪便排出,幼虫体内的毒蛋白含量一直低于粪便中的含量,且在24 h时两者差异达到最大。由于Bt毒蛋白在幼虫体内的积累,扰乱了稻纵卷叶螟幼虫体内SOD,CAT和POD 3种保护酶的动态平衡,使虫体内自由基的清除遇到障碍,从而对其产生毒害作用。  相似文献   

8.
转cry1Ab基因水稻对非靶标昆虫白背飞虱种群增长的影响   总被引:1,自引:0,他引:1  
周霞  程家安  娄永根 《昆虫学报》2006,49(5):786-791
通过室内饲养实验及水稻氨基酸和碳、氮含量测定,比较研究了转cry1Ab水稻克螟稻 (KMD1和KMD2)与其亲本秀水11对非靶标害虫白背飞虱种群增长的影响。 结果表明,以克螟稻为食可对白背飞虱的产卵期和每雌产卵量产生一定影响。 以KMD2为食的白背飞虱的产卵期为7.6天,每雌产卵量为95.0粒,均显著低于以母本秀水11为食的白背飞虱的12.7天和167.5粒。但是,以KMD1为食的白背飞虱仅产卵期(8.6天)显著变短, 其每雌产卵量与以其母本秀水11为食的白背飞虱无显著差异。稻苗氨基酸含量分析结果表明KMD2引起白背飞虱种群生殖力显著下降可能与其游离氨基酸的总量和丙氨酸含量的显著下降以及谷氨酸含量的显著上升有关。因此,转cry1Ab水稻可对白背飞虱种群增长产生影响,但其影响因转cry1Ab水稻品种而异。  相似文献   

9.
Bt水稻Cry1Ab杀虫蛋白表达的时间动态及其在水稻土中的降解   总被引:11,自引:0,他引:11  
白耀宇  蒋明星  程家安 《生态学报》2005,25(7):1583-1590
采用ELISA技术检测了Bt水稻克螟稻1号(KMD1)和克螟稻2号(KMD2)不同生育期茎和叶片中cry1Ab基因的表达水平,以及植株组织(茎和叶片)室内埋于不同稻田土壤中其Cry1Ab杀虫蛋白的降解动态。结果表明,不论是KMD1还是KMD2,茎和叶片中cry1Ab基因的表达量为3.7~9.1μg/g鲜重,均以拔节期和孕穗期相对较低,灌浆初期显著升高,黄熟期又明显下降(但仍然维持在较高的表达量)。KMD1茎和叶片中Cry1Ab蛋白在3种水稻土即青紫泥田、黄松田和黄筋泥田中的降解,均以前期(处理后12d内)较快,但中后期明显较慢;处理后42d,这两种组织中的Cry1Ab残留量均稳定在μg级水平,分别仅有0.20~0.85μg/g干重和0.35~1.81μg/g干重。KMD1茎和叶片中Cry1Ab蛋白降解动态与土壤类型密切相关,青紫泥田中降解最快,黄筋泥田中次之,黄松田中最慢。淹水可加快土壤中茎和叶的腐解,从而促进其中Cry1Ab蛋白的降解。KMD2粉碎叶中Cry1Ab蛋白在有菌水稻土中的降解快于无菌土中,表明土壤微生物存在可加快Cry1Ab蛋白的降解。在各种土壤淹水与非淹水、灭菌与非灭菌处理条件下的Cry1Ab蛋白降解过程均可用指数方程进行拟合,算得其消减半衰期为2.2~11.6d不等。讨论了土壤类型、淹水等影响Cry1Ab蛋白降解的可能机制,认为土壤微生物是其中起关键作用的一个因子。  相似文献   

10.
分离和鉴定二化螟Chilo suppresalis幼虫中肠刷状缘膜囊泡(BBMV)中Cry1A毒素的受体蛋白,对于阐明Cry1A毒素作用机理和二化螟抗性机理具有十分重要的意义。为此,本文就Cry1A毒素对二化螟杀虫活性及Cry1Ac与二化螟中肠受体的配基结合进行了研究。结果表明: Cry1Ab对二化螟室内品系(CN)的毒力高于Cry1Ac,而Cry1Ac高于Cry1Aa。配基结合分析表明二化螟CN品系幼虫中肠BBMV中有6个Cry1Ac结合蛋白(分子量分别为50,70,90,120,160和180 kDa), 其中180,160和90 kDa结合蛋白的条带颜色明显深于其他结合蛋白的条带,表明这3个受体蛋白具有较高的结合浓度。同源竞争结合研究表明,180和90 kDa结合蛋白为Cry1Ac的低亲合性结合蛋白,其他4个为高亲合性结合蛋白。为了研究Cry1Ac和Cry1Ab受体结合部位的相互作用,进行了异源竞争结合研究。Cry1Ab可以与Cry1Ac所有的6个结合蛋白进行竞争性结合,与180,120,70和50 kDa结合蛋白具有高亲合性,而与160和90 kDa结合蛋白具有低亲合性。结果显示,Cry1Ac与Cry1Ab在二化螟幼虫中肠BBMV上拥有多个共享的结合位点,但对每个结合位点的亲合性有差异。基于毒素结合部位的相似性,Cry1Ac和Cry1Ab不宜同时用于转基因Bt水稻来控制二化螟。  相似文献   

11.
通过开顶式气室研究了CO2浓度倍增(750vs.375μL/L)对取食转Bt水稻及其对照亲本秀水11的二化螟Chilo suppressalis(Walker)幼虫体内代谢酶乙酰胆碱酯酶(AChE)和保护酶(超氧化物歧化酶SOD、过氧化物酶POD和过氧化氢酶CAT)活力的影响,以期明确该类害虫对大气CO2浓度升高及转Bt水稻的生理响应。研究结果表明:(1)AChE受CO2浓度水平影响显著,与对照CO2处理相比,高CO2条件下该酶活力显著降低;(2)高CO2水平及水稻中Bt毒素蛋白的存在均会导致二化螟幼虫体内POD活力显著降低,SOD初期活力显著升高,但不能维持较长时间,CAT活力表现为先升后降的特点;(3)当受到水稻品种、CO2浓度水平和取食时间的综合影响时,以上代谢酶和保护酶活力会发生显著变化。短时间内二化螟幼虫可通过体内以上生理代谢调节减缓受CO2浓度升高和水稻中Bt毒素蛋白的不利影响,但长时间环境胁迫下这种应对不利环境的响应机制将会失去作用,最终导致种群适合度的降低。  相似文献   

12.
One major concern regarding the release of Bt rice is its potential impact through tritrophic interactions on nontarget arthropods, especially natural enemies. We studied the effects of two Bt transgenic rice varieties, TT9- 3 and KMD1, expressing Cry1Ab/Cry1Ac and Cry1Ab, respectively, on a predatory ground spider [Ummeliata insecticeps (B?senberg et Strand)] supplied with Bt rice-fed brown planthopper [Nilaparvata lugens (St?l)] nymphs. Although immunoassays confirmed that U. insecticeps ingested Bt insecticidal protein when supplied with Bt rice-fed N. lugens, no negative effects were found on its survival and development. Furthermore, the fecundity of U. insecticeps fed prey reared on Bt rice was not significantly different from that of those fed prey reared on non-Bt rice. A 3-yr field trial indicated that Bt rice did not significantly affect the population density of U. insecticeps in comparison with non-Bt rice. In conclusion, the Bt rice lines tested in this study had no adverse effects on the survival, developmental time, or fecundity of U. insecticeps in the laboratory or on population dynamics in the field.  相似文献   

13.
The receptor binding step in the molecular mode of action of five delta-endotoxins (Cry1Ab, Cry1Ac, Cry1C, Cry2A, and Cry9C) from Bacillus thuringiensis was examined to find toxins with different receptor sites in the midgut of the striped stem borer (SSB) Chilo suppressalis (Walker) and yellow stem borer (YSB) Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae). Homologous competition assays were used to estimate binding affinities (K(com)) of (125)I-labelled toxins to brush border membrane vesicles (BBMV). The SSB BBMV affinities in decreasing order was: Cry1Ab = Cry1Ac > Cry9C > Cry2A > Cry1C. In YSB, the order of decreasing affinities was: Cry1Ac > Cry1Ab > Cry9C = Cry2A > Cry1C. The number of binding sites (B(max)) estimated by homologous competition binding among the Cry toxins did not affect toxin binding affinity (K(com)) to both insect midgut BBMVs. Results of the heterologous competition binding assays suggest that Cry1Ab and Cry1Ac compete for the same binding sites in SSB and YSB. Other toxins bind with weak (Cry1C, Cry2A) or no affinity (Cry9C) to Cry1Ab and Cry1Ac binding sites in both species. Cry2A had the lowest toxicity to 10-day-old SSB and Cry1Ab and Cry1Ac were the most toxic. Taken together, the results of this study show that Cry1Ab or Cry1Ac could be combined with either Cry1C, Cry2A, or Cry9C for more durable resistance in transgenic rice. Cry1Ab should not be used together with Cry1Ac because a mutation in one receptor site could diminish binding of both toxins.  相似文献   

14.
Zhang Y  Li Y  Zhang Y  Chen Y  Wu K  Peng Y  Guo Y 《Environmental entomology》2011,40(5):1323-1330
Laboratory bioassays and field surveys were carried out to compare the resistance of three transgenic rice (Oryza sativa L.) lines including Bt-DL expressing a single gene cry1Ab, Bt-KF6 expressing stacked genes cry1Ac and CpTI genes and Bt-SY63 expressing a fusion gene cry1Ab/cry1Ac, respectively, to an important rice pest Chilo suppressalis (Walker). In addition, enzyme-linked immunosorbent assays (ELISA) were conducted to monitor the Bt protein expressions in rice leaves and stems at different rice growth stages. Results showed that all the transgenic rice lines exhibited significantly high resistance to the pest compared with their corresponding nontransformed isolines. Among the transgenic rice lines, Bt-SY63 and Bt-KF6 had higher resistance to C. suppressalis at early growth stage, but lower resistance at late stages, while the pest resistance of Bt-DL was relatively stable throughout the growing season. The results were consistent with ELISA results showing that Bt protein levels in Bt-SY63 or Bt-KF6 leaves decreased in late growth stages, but were relatively stable in Bt-DL at all growth stages. This demonstrates that the resistance to a pest by Bt plants is positively correlated with Cry protein expression levels in plant tissues. Compared with Bt-SY63 and Bt-KF6, the Bt protein expression levels were significantly lower in Bt-DL, while its resistance to C. suppressalis was the highest. This may suggest that C. suppressalis is more susceptible to Cry1Ab than to Cry1Ac. The data from the current study are valuable for decision-making for commercial use of Bt rice lines and development of appropriate pest control and resistance management strategies for the transgenic rice lines.  相似文献   

15.
Heritability of Chilo suppressalis (Walker) tolerance to the Cry1Ab toxin of Bacillus thuringiensis Berliner was estimated using a half-sibling design. Artificial diet with and without Cry1Ab was infested with progenies of 20 males, each mated with 2 females, and mortality was scored 5 d after infestation. The progeny of each female was reared and scored separately. Mean mortality of the 20 families on the Cry1Ab diet was 46.5%. The effects of both male parent and of female parent within male parent were significant. Heritability was estimated to be 0.52, suggesting that a high proportion of phenotypic variation was because of genetic differences. Mortality on the Cry1Ab diet was not correlated with mortality on control diet, indicating that differences among families in tolerance to Cry1Ab were not attributable to differences in general fitness. Our results indicate that "high dose" Bt rice plants may be particularly important for Cry1Ab resistance management in C. suppressalis populations.  相似文献   

16.
在实验室中以转cry1Ab基因水稻 克螟稻 1号为材料 ,研究了不同温度下Bt水稻对二化螟Chilosuppressalis (Walker) 3龄和 5龄幼虫取食、生长及其存活的影响。结果表明 ,不同温度下 3龄和 5龄幼虫取食Bt水稻后 ,其取食量、体重增长和存活率均极显著低于对照。温度对 3龄幼虫取食、生长和存活无显著影响 ,但对 5龄幼虫的取食和体重增长则有显著影响。幼虫死亡率与其取食Bt水稻的累积食量间存在着正相关关系。  相似文献   

17.
转Bt基因水稻对两种弹尾虫及尖钩宽黾蝽捕食作用的影响   总被引:6,自引:0,他引:6  
转Bt基因水稻KMD1、KMD2和对照水稻XS11稻田主要有两种弹尾虫:灰橄榄长角跳虫 Entomobryagriseoolivata (Packard) 和钩圆跳虫 Bourletiella christianseni Banks。两种Bt稻田中灰橄榄长角跳虫种群密度均显著高于对照XS11稻田;在以KMD1和KMD2腐烂 茎叶为食的灰橄榄长角跳虫成虫中检测到微量Cry1Ab杀虫蛋白。室内测定结果表明,不管是单 头捕食还是多头协同捕食,尖钩宽黾蝽Microvelia horvathi Lundblad 成虫对用3种供试 水稻残体饲养的灰橄榄长角跳虫的捕食量和功能反应均符合HollingⅡ型方程,其日捕食量、瞬 时攻击率(a)和处理时间(Th)均无显著差异。  相似文献   

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