转Cry1Ac+CpTI基因棉对棉田害虫及其天敌种群动态的影响

002年在河北省南皮县对转Cry1Ac+CpTI基因棉（SGK321）棉田害虫及其天敌种群动态的研究结果表明,SGK321棉田及其亲本对照棉（石远321）棉田的害虫和捕食性天敌的种类基本相同,但数量差异较大。但在5月23日至9月16日的24次调查中,SGK321棉田的5 种主要害虫棉铃虫、棉蚜、绿盲蝽、棉粉虱、小绿叶蝉的总数量分别较其亲本石远321棉田降低89.5%、64.5%、21.8%、15.6%和33.7%。SGK321棉田龟纹瓢虫和中华草蛉的种群总数量分别比石远321棉田增高34.0%和9.1%,但异色瓢虫、小花蝽、异须盲蝽、蚜茧蜂和蜘蛛类的种群数量分别降低28.6%、6.5%、43.1%、44.7%和14.0%。主要害虫和天敌种群动态的监测表明,棉蚜、小绿叶蝉和棉粉虱的发生高峰期分别为7月中下旬,8月下旬至9月中旬,8月下旬至9月上、中旬。在三者的高峰期内,SGK321棉田的种群数量基本上低于对照田。龟纹瓢虫的发生高峰期为7月上旬到8月上、中旬,且SGK321棉田的种群数量高于对照田。研究表明, SGK321在对棉铃虫具有很好抗性的同时,对棉蚜、棉粉虱、绿盲蝽、小绿叶蝉等非靶标害虫的发生也有一定的抑制作用;SGK321棉田龟纹瓢虫和中华草蛉的种群数量增加,其他主要天敌的数量则有所降低,表明SGK321对某些天敌种类的种群动态存在不利影响。     

Effect of cotton aphids feeding on transgenic cotton varieties on the behavior of Propylaea japonica

本文以GK12、33B和SGK3213种转基因棉和对应的3种常规棉泗棉3号、33及石远321为材料,通过室内测定,研究了取食转基因棉的棉蚜Aphis gossypii Glover对龟纹瓢虫Propylaea japonica Thunberg1～4龄幼虫功能反应的影响.结果指出,龟纹瓢虫对来自转基因棉田蚜虫的捕食量大多高于常规棉田,且捕食量随猎物密度的增加而增大.在1、2龄幼虫期,龟纹瓢虫取食转双价基因棉田蚜虫的数量多高于取食转单价基因棉田的蚜虫数量,但是在3、4龄幼虫时,龟纹瓢虫取食转双价棉田蚜虫的数量多低于取食转单价棉田的蚜虫数量.龟纹瓢虫对来自转基因棉田棉蚜的捕食功能反应符合HollingⅡ型.龟纹瓢虫1龄幼虫取食GK12和SGK321 2个转基因棉田蚜虫的最大日捕食量大于对照,处理时间短于对照;龟纹瓢虫P.japonica 2、3龄幼虫取食33B和SGK321 2个转基因棉田蚜虫A.gossypii的日最大捕食量大于对照,处理时间短于对照;4龄时取食转基因棉田蚜虫的数量都大于对照,处理时间均短于对照,其中取食转双价基因棉SGK321棉田蚜虫的日最大捕食量小于取食单价棉田蚜虫的日最大捕食量.     

取食不同品种棉花的棉蚜对龟纹瓢虫功能反应的影响

本文以GK12、33B和SGK3213种转基因棉和对应的3种常规棉泗棉3号、33及石远321为材料,通过室内测定,研究了取食转基因棉的棉蚜却h／sgossypiiGlover对龟纹瓢虫Propylaea japonica Thunberg1—4龄幼虫功能反应的影响。结果指出,龟纹瓢虫对来自转基因棉田蚜虫的捕食量大多高于常规棉田,且捕食量随猎物密度的增加而增大。在1、2龄幼虫期,龟纹瓢虫取食转双价基因棉田蚜虫的数量多高于取食转单价基因棉田的蚜虫数量,但是在3、4龄幼虫时,龟纹瓢虫取食转双价棉田蚜虫的数量多低于取食转单价棉田的蚜虫数量。龟纹瓢虫对来自转基因棉田棉蚜的捕食功能反应符合HollingII型。龟纹瓢虫1龄幼虫取食GK12和SGK3212个转基因棉田蚜虫的最大日捕食量大于对照,处理时间短于对照;龟纹瓢虫P．japonica2、3龄幼虫取食33B和SGK3212个转基因棉田蚜虫A．gossypii的日最大捕食量大于对照,处理时间短于对照;4龄时取食转基因棉田蚜虫的数量都大于对照,处理时间均短于对照,其中取食转双价基因棉SGK321棉田蚜虫的日最大捕食量小于取食单伊棉田蚜虫的日最大捕食量。     

转Bt基因抗虫棉(Gossyposium)伤流中Bt毒蛋白的运输

本文以常规棉品种‘石远321'和抗虫棉品种‘SGK321'与‘99B'为材料,通过考马斯亮蓝和ELISA方法分析了伤流中可溶性蛋白和Bt(苏云金芽孢杆菌)毒蛋白含量.结果发现,转基因抗虫棉伤流中可溶性蛋白含量显著高于常规棉,并在抗虫棉伤流中检测到了Bt毒蛋白,而常规棉伤流中没有Bt毒蛋白的存在.通过嫁接实验研究发现,将常规棉嫁接到转Bt基因抗虫棉砧木上,叶片中也有Bt毒蛋白的积累.这些结果说明,Bt毒蛋白可以通过木质部伤流液向地上部运输,并在叶片中积累,Bt毒蛋白由根系向地上部的运输对棉花的抗虫性是有益的.     

转双价基因抗虫棉对土壤微生物群落多样性的影响

采用Biolog技术,研究了不同生育期(30、60、90和120d)转双价(Bt+CpTI)基因抗虫棉SGK321及其亲本非转基因常规棉石远321根际土壤微生物群落多样性变化。结果表明,反映土壤微生物活性的平均颜色变化率(AWCD)呈现出以下变化规律:与亲本常规棉相比,整个培育过程中转双价基因抗虫棉AWCD值除30d显著高于亲本常规棉(P<0.05),其他3个时期均显著低于亲本常规棉(P<0.05)。两种棉花根际土壤微生物群落丰富度指数和优势度指数随棉花生育期的不同而有所不同,4个时期转双价基因抗虫棉均匀度指数与亲本常规棉无显著差异。主成分分析结果表明,转双价基因抗虫棉30d、亲本常规棉60、120d土壤微生物群落碳源利用方式相似,转双价基因抗虫棉60、90、120d与亲本常规棉30d土壤微生物群落碳源利用方式相似。土壤微生物利用的主要碳源为糖类、氨基酸类、羧酸类和聚合物。     

取食转Bt+CpTI基因棉花的棉蚜对普通草蛉生长发育及繁殖力的影响

【目的】研究取食转Bt+CpTI基因棉花的棉蚜Aphis gossypii Glover对普通草蛉Chrysopa carnea Stephens生长发育及繁殖力的影响,为转Bt基因抗虫棉花的推广和应用提供理论支持。【方法】普通草蛉取食转Bt+CpTI棉花SKG321和对照品种石远321棉花上的棉蚜后,对普通草蛉生长发育情况进行研究,并对其幼虫、蛹的存活率和成虫的羽化率及产卵量进行分析。【结果】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的各龄幼虫发育历期、蛹期和产卵前期无不利影响;处理与对照间各龄幼虫存活率、蛹的存活率和成虫羽化率之间没有显著差异;取食转Bt+CpTI棉花上棉蚜的普通草蛉单雌日产卵量与对照相比没有减少,而25 d内的单雌总产卵量与对照相比差异亦不显著。【结论】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的生长发育及繁殖力无负面影响。     

取食转Bt基因棉花上的棉蚜对丽草蛉发育和繁殖的影响

为明确转Bt基因抗虫棉对捕食性天敌的生态效应 ,比较了 2种转Bt基因抗虫棉 (GK-1 2和新棉99B)和 1种常规棉上的棉蚜Chrysopaformosa对丽草蛉Aphisgossypii发育和繁殖的影响。结果表明 ,(1 )与取食常规棉泗棉 3号上棉蚜的对照相比 ,取食转Bt基因抗虫棉GK- 1 2上棉蚜的第 1代和第 2代丽草蛉 ,其幼虫期和茧期的死亡率、幼虫和茧的发育历期、茧重及成虫性比等均与对照无显著差异 ;但第 1代成虫的产卵量减少了 2 88 0粒 ,与对照差异显著 (P <0 . 0 1 ) ;第 2代成虫所产卵的孵化率为 64. 0 % ,显著低于对照的 77. 7% (P <0 0 1 )。 (2 ) 2个品种的Bt棉对丽草蛉发育和繁殖的影响也略有差异。与取食GK 1 2上棉蚜的个体相比 ,取食新棉 99B上蚜虫的第 1代丽草蛉 ,其幼虫发育历期缩短了 0 . 6d(P <0 0 1 ) ,茧期缩短了 0 . 7d(P <0 . 0 1 ) ,茧重降低了 1 . 2mg(P <0 .0 1 ) ,幼虫期和茧期的死亡率、茧重以及成虫性比等则无显著差异 ;雌成虫产卵前期、产卵期、产卵量和寿命等繁殖学特性也无显著差异 ,但其成虫所产卵的孵化率为 65. 0 % ,显著低于取食GK- 1 2上棉蚜的处理 (72 . 7% ,P <0. 0 1 )。分别取食 2种Bt棉上的棉蚜 ,对第 2代丽草蛉的发育和繁殖的影响差异则较小。     

转Bt 基因抗虫棉(Gossyposium)伤流中Bt 毒蛋白的运输

本文以常规棉品种‘石远321’和抗虫棉品种‘SGK321’与‘99B’为材料, 通过考马斯亮蓝和ELISA方法分析了伤流中可溶性蛋白和Bt(苏云金芽孢杆菌)毒蛋白含量。结果发现, 转基因抗虫棉伤流中可溶性蛋白含量显著高于常规棉, 并在抗虫棉伤流中检测到了Bt毒蛋白, 而常规棉伤流中没有Bt毒蛋白的存在。通过嫁接实验研究发现, 将常规棉嫁接到转Bt基因抗虫棉砧木上, 叶片中也有Bt毒蛋白的积累。这些结果说明, Bt毒蛋白可以通过木质部伤流液向地上部运输, 并在叶片中积累, Bt毒蛋白由根系向地上部的运输对棉花的抗虫性是有益的。     

湖北棉区转Bt基因棉对棉铃虫的控制作用

2000-2001年通过田间系统调查,表明转Bt基因棉（品种为GK19）在湖北江汉平原棉区对棉铃虫抗性稳定。试验设3个处理：转Bt基因棉化防田（使用化学农药控制害虫）、转Bt基因棉自控田（依靠天敌控制害虫）及常规棉对照田（利用综合防治措施控制害虫）。从棉铃虫的第2代到第5代整个发生期内,即使在不进行化学防治的情况下,棉铃虫在Bt棉田的发生量也保持在极低的水平(最高百株虫量为12头)。室内饲养结果表明,转Bt基因棉对棉铃虫的生长发育（幼虫体重、蛹重）有较为明显的影响,使6龄幼虫体重减少25.6%,蛹重减少18.2%。棉铃虫幼虫取食转Bt基因棉组织后,发育迟缓,相对于常规棉喂养的整个发育历期延长17 d,使棉铃虫在田间的危害减少至少一个世代。另外,接虫试验表明,棉铃虫幼虫在常规棉上的取食时间是转Bt基因棉株上的6.1倍,极大地减轻了棉铃虫的危害程度。     

转Bt基因棉花对棉铃虫不同龄期幼虫的杀虫活性和抑制生长作用

以我国培育的转Bt基因棉花(简称Bt棉)为材料,系统测定了Bt棉叶片对棉铃虫 Helicoverpa armigera(Hubner)不同龄期幼虫的杀虫活性和抑制生长的作用。结果表明,Bt棉的杀虫活性随着龄期的增大而趋于降低,其中处理3d只对1龄幼虫有较高效果;从1～4龄开始连续取食Bt棉后均不能化蛹,5龄幼虫则能正常化蛹。3龄幼虫取食Bt棉叶3d后的体重与初始体重之比为0.94,而取食常规棉叶的相应比值为5.48,对幼虫生长的抑制作用明显。在28℃条件下用Bt棉饲养棉铃虫1～3龄幼虫3d,对幼虫的致死率显著高于25℃处理,对3龄幼虫抑制生长的作用也显著提高。研究结果可为确定棉铃虫对Bt棉的抗性监测与治理技术以及Bt棉的田间应用技术提供依据。     

Feeding behaviour of Helicoverpa armigera larvae on insect-resistant transgenic cotton and non-transgenic cotton

Abstract: Feeding behaviour of Helicoverpa armigera Hübner (Lep.; Noctuidae) larvae on non‐transgenic Bacillus thuringiensis (Bt) cotton (Gossypium hirsutum L.), Zhong 30, and transgenic cowpea trypsin inhibitor (CpTI)‐Bt cotton, SGK 321, and non‐transgenic cotton, Shiyuan 321, was investigated in both choice tests and no‐choice tests. The results of choice tests suggested that neonates have the ability to detect and avoid transgenic cotton. In the choice tests of neonates with both transgenic and non‐transgenic cotton leaves, a significantly greater proportion of larvae and higher consumption were observed on non‐transgenic cotton than on the transgenic Bt or CpTI‐Bt cotton. In the choice tests with leaves of two transgenic cotton lines, the proportion of neonates on leaf discs of the two lines was not significantly different, but there was significantly higher consumption on CpTI‐Bt transgenic cotton than that on Bt transgenic cotton. In addition, significantly more neonates were found away from the leaf discs, lower consumption and higher mortality were achieved in the choice test with two transgenic cotton leaves than in the choice tests containing non‐transgenic cotton leaves. Leaves and buds were examined in choice tests of fourth instars. It appeared that fourth instars were found in equal numbers on transgenic and non‐transgenic cotton, except when larvae were exposed to leaves for 3 h. However, the total consumption on transgenic cotton was lower than that of the non‐transgenic cotton, so fourth instars may still have the capacity to detect transgenic cotton and reduce feeding on it, although they showed no preference on either transgenic or non‐transgenic cotton. More larvae were found off diet in the treatments with leaves than that of buds, and the number of injured leaf discs by per fourth instar was significantly higher than that of buds in choice tests, suggesting that leaf is a less preferred organ for H. armigera larvae, elicited more larval movements. Similarly, in no‐choice tests of fifth instars, significantly fewer feeding time and more moving time occurred on leaf than that of bud, boll and petal. When cotton line was considered, compared with non‐transgenic cotton, significantly lower feeding time and higher resting time occurred on the two transgenic cottons. Overall, H. armigera larvae have the ability to detect the transgenic Bt and CpTI‐Bt cottons or the less preferred organs and selectively feed more on the non‐transgenic cotton or the preferred organs, especially the neonates, which have a high capacity for avoiding transgenic cotton.     

Effects of repeated cultivation of transgenic Bt cotton on functional bacterial populations in rhizosphere soil

The impact of multiple-year (0–5 years) cultivation of transgenic Bacillus thuringiensis (Bt) cotton on the functional bacterial populations in rhizosphere soil was investigated. The transgenic Bt + CpTI cotton line SGK321 and a non-Bt cotton line Shiyuan321 were planted in four fields in which Bt cotton had been continuously cultivated for 0, 1, 3, and 5 years. Rhizosphere soil samples were collected at the seedling, squaring, flower and boll, and boll-opening stages of cotton. Numbers of bacteria involved in nitrogen-fixing, organic phosphate-dissolving, inorganic phosphate-dissolving, and potassium-dissolving were measured with cultivation-dependent approaches. The data presented here showed no consistent statistically significant differences in the numbers of different groups of functional bacteria between rhizosphere soil of Bt and non-Bt cotton in the same field, and no obvious trends in the numbers of the various group of functional bacteria with the increasing duration of Bt cotton cultivation. These studies suggest that multiple-year cultivation of transgenic Bt cotton may not affect the functional bacterial populations in rhizosphere soil.     

Impact of transgenic Bacillus thuringiensis cotton on a non-target pest Tetranychus spp. in northern China

Field studies were carried out to evaluate the effect of two transgenic cotton varieties (SGK321 carrying Cry1A+CpTI and DP99B carrying Cry1Ac) and the conventional variety (shiyuan321‐parental line of SGK321) on spider mites, Tetranychus spp. from 2002 to 2004. In 2002, this experiment included three treatments: Bt cotton field (SGK321) treated with acaricides against spider mites, untreated non‐Bt cotton field (Shiyuan321), and untreated Bt cotton field (SGK321). In 2003–2004, there are four types of treatments after a new transgenic Bt cotton variety, DP99B (non‐chemical control), was added into the experiments. The results showed that there were no significant difference in densities of spider mites among Bt without acaricides and non‐Bt without acaricides cotton fields, nor was there a significant difference in damage of spider mites to cotton among these treatments (P > 0.1). However, there are significant differences (P < 0.05) in densities of spider mites and damage caused by spider mites between cotton fields with and without acaricides. Acaricide significantly reduced the densities of spider mites in Bt cotton (P < 0.05). These results suggest that Bt cotton has no effect on spider mites populations. However, spider mites have the potential for severe damage in Bt cotton fields. Acaricides are essential tools in controlling cotton spider mites in northern China.     

转Bt基因抗虫棉对棉铃虫拒食作用及其机理研究

转Bt基因抗虫棉对 3 ,4,5龄棉铃虫幼虫的抗性表现形式为拒食作用 ,且随幼虫龄期的增加拒食作用明显降低 ,其中对 5龄幼虫的拒食作用很低。取食Bt棉后 ,3 ,4,5龄棉铃虫幼虫中肠消化酶比活力均较对照有所减退 ,且随幼虫龄期的增加减退率明显降低 ,其中对 5龄幼虫减退率最低。由此 ,解释了Bt棉对 3龄及 3龄以上棉铃虫幼虫抗性表现形式、抗性随不同幼虫龄期的差异性 ,及其抗性差异性的消化机理。     

转基因抗棉铃虫抗草甘膦棉花对棉铃虫的抗虫性及对二种地老虎幼虫生长发育的影响

为了明确转Bt Cry1A+CP4EPSPS基因抗棉铃虫抗草甘膦棉花对棉铃虫Helicoverpa armigera(Hübner)的抗性水平及对八字地老虎Amathes c-nigrum和黄地老虎Agrotis segetum的影响,作者用转基因棉花和非转基因棉花叶片对棉铃虫、八字地老虎和黄地老虎幼虫进行了室内饲养观察。结果表明,转基因抗棉铃虫抗草甘膦棉花对新疆南部第2代棉铃虫表现为中抗,幼虫校正死亡率为70.4%;棉花叶片在整个生育期均能表达Bt毒蛋白,整个生育期Bt毒蛋白含量呈逐渐降低的趋势。转基因抗棉铃虫抗草甘膦棉花对八字地老虎幼虫有强烈的毒杀作用,可以引起幼虫大量死亡,对存活幼虫的生长发育也有明显的抑制作用,幼虫发育迟缓,不能顺利化蛹;但对黄地老虎幼虫没有明显的影响,幼虫可以正常生长。     

转基因棉花对棉铃虫天敌寄生率的影响

采用分期接虫、分期回收法研究常规棉与转基因棉花品种 (系 )上棉铃虫寄生性天敌在不同棉花品种上的寄生作用。结果表明 ,无论是第 2代棉铃虫发生期间还是第 3 ,4代棉铃虫发生期间 ,无论是棉铃虫卵期还是幼虫期 ,转基因棉花品种 (系 )的棉铃虫卵、幼虫寄生率均显著低于常规棉花品种上的寄生率。棉铃虫卵期寄生蜂主要是拟澳洲赤眼蜂 ,幼虫寄生蜂为棉铃虫齿唇姬蜂等。由此看出转基因棉对棉铃虫寄生性天敌存在非亲和性。     

转基因棉Bt毒蛋白含量与抗棉大卷叶螟效果间的关系分析

棉大卷叶螟Sylepta derogata Fabricius为近年来长江流域棉花中后期的一种重要害虫。在其危害盛期测定了8种转基因棉叶片Bt毒蛋白的含量与受害程度,在此基础上就转基因棉对棉大卷叶螟的抗虫性采用不同抗性指标相结合的方法进行了综合评估,同时对转基因棉对棉大卷叶螟的抗虫效果与毒蛋白含量的相关性进行分析。结果表明:各转基因棉品种的不同部位叶片的毒蛋白含量呈现顶叶最高,功能叶次之,老叶含量最低的趋势。抗蚜8017和SGK321 2个转基因棉品种的棉叶毒蛋白表达量相对较低,平均值在120μg/g以下,对棉大卷叶螟的抗性级别均为中抗。其它供试转基因棉棉叶毒蛋白表达量均值在150μg/g以上,抗性级别也均为高抗,而非转基因棉泗棉3号与石远321对棉大卷叶螟不具抗性。转基因棉对棉大卷叶螟的抗性水平与毒蛋白含量呈正相关。     

新疆棉区转Bt基因棉对棉铃虫生物学的影响

棉铃虫Helicoverpaarmigera高龄幼虫取食转Bt基因棉花组织后 ,化蛹率、羽化率、蛹重、体长均有显著下降 ,在转Bt基因棉花上棉铃虫的取食行为也受到较大的影响 ,表现为取食次数明显减少、吐丝下垂次数明显增加 ;无论是转Bt基因棉花还是常规棉花 ,棉铃虫 3龄幼虫主要分布于繁殖器官上 ,在转Bt基因棉花各繁殖器官上的分布概率为 :花 >棉铃 >棉蕾 ,常规棉花上分布概率为 :棉蕾 >棉铃 >花 ;棉铃虫高龄幼虫取食转Bt基因棉花各组织 ,成虫羽化后产卵量、卵孵化率均有明显下降 ;在较低的棉铃虫虫口密度下 ,转Bt基因棉花对棉铃虫有一定的产卵排趋性。     

外源基因导入对棉花烟粉虱种群的影响

以转基因棉花和对应的常规棉花亲本为材料,探讨外源基因导入对棉花烟粉虱Bemisia tabaci(Gennadius)种群的影响。结果表明,在转基因棉花GK12、33B、SGK上,烟粉虱从卵发育到成虫羽化的历期分别为15.56、15.35和15.25 d,较对应的常规棉亲本SM3(19.38 d)、33(20.81 d)、SY321(18.76 d)分别短24.55%、26.23%、18.71%;GK12和33B棉花上烟粉虱的存活率(69.16%)分别较对应的常规棉亲本SM3(54.76%)和33(64.91%)高26.29%和12.81%,而SGK(63.21%)上烟粉虱的存活率与对应的常规棉亲本SY321(62.61%)之间差异不显著;烟粉虱在GK12(84.00)和33B(77.25)上的产卵量分别较SM3(62.25)、33(70.00)高34.93%和10.35%,但SGK和SY321之间差异不显著;在转基因棉花GK12、33B、SGK上,烟粉虱的雌雄性比分别比对应的常规棉亲本高26.71%、46.23%和19.17%。结果表明,外源基因导入后,有助于烟粉虱的发育,提高了烟粉虱的产卵量和雌雄性比,从而促进了烟粉虱种群的上升。