【作者】 姜永厚；

【导师】 程家安； 张志涛；

【作者基本信息】 浙江大学 ， 农业昆虫与害虫防治， 2004， 博士

【摘要】 本文在室内以转Bt基因水稻(KMD1)和转Bt／Sck双价水稻(MSB)两种转基因抗虫水稻——两种水稻主要害虫(二化螟Chilo suppressalis、褐飞虱Nilaparvata lugens)和一种偶发性害虫(稻眼蝶Mycalesis gotama)——三种主要天敌(二化螟绒茧蜂Apanteles chilonis、尖钩宽黾蝽Microvrlia horvathi和拟水狼蛛Pirata subpiraticus)为研究系统，研究了转基因抗虫水稻经害虫对天敌生物学特性的影响及可能的作用机理，实验结果如下： 一、转Bt基因水稻对二化螟的存活和生长以及二化螟绒茧蜂生物学特性的影响 连续取食KMD1的三、四、五龄二化螟的校正死亡率都在2～4天内迅速上升，分别达90.1％、80.3％、45.7％：至第6天和第7天三、四龄二化螟的校正死亡率达100％；至第8天五龄二化螟达96％。三、四、五龄二化螟在KMD1取食不同时间后至对照品种继续取食，其校正死亡率随在KMD1取食时间的延长而上升。 五龄二化螟取食KMD1不同时间后至对照品种继续取食一段时间，体重均低于对照，其中处理后当天和3天差异达极显著水平，处理后9天差异仍达显著水平。 二化螟绒茧蜂寄生取食过KMD1的二化螟后，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、茧块大小、蜂羽化率及性比均无显著影响。 二、转cry1Ac+sck双价抗虫水稻对二化螟存活和生长以及二化螟绒茧蜂生物学特性的影响 连续取食MSB后二化螟的死亡率有所上升，从第6天开始，取食MSB的二化螟的死亡事极显著高于取食对照明恢86的死亡率。从第2天开始，取食MSB的二化螟幼虫体重显著低于取食明恢86对照的。 取食MSB 36h后至明恢86对照上饲养0h、24h、48h、72h后，二化螟的死亡率均高于相应的对照，但差异都不显著，但其体重均极显著低于相应的对照。 取食MSB 36h后至明恢86对照上饲养3天、6天、9天、12天后，二化螟的死亡率均高于相应的对照，但差异都不显著；而二化螟体重均低于对照，其中3天和6天差异分别达极显著和显著水平。 二化螟绒茧蜂寄生取食过MSB的三、四、五龄二化螟，寄生率都低于对照，其中四龄差异达显著水平；三龄、四龄和五龄二化螟的结茧率与对照差异都不显著；寄生在取食MSB的五龄二化螟幼虫体内的二化螟绒茧蜂蛹期显著长于对照，而所结茧的茧长显著短于对照。但是，卵+幼虫历期、茧块大小、羽化率、雌性比、雌雄蜂寿命和前翅长与对照无明显差异。浙江大学博士学位论文三、取食转基因抗虫水稻后二化螟幼虫血淋巴中游离氨基酸含量的变化 五龄二化螟幼虫取食KMDI或双价基因水稻36h后，血淋巴中氨基酸总量与对照相比，都发生一些变化。其中，取食KMI)1后O天和3天氨基酸总量均显著高于对照;取食MSB后O天、1天和3天氨基酸总量与对照无显著差异。 取食转基因水稻后血淋巴氨基酸种类与对照相比虽未发生变化，但在数量上发生不同程度的变化。取食KMDI 36h后O天，20种氨基酸中除天门冬酸胺、丝氨酸、苏氨酸和甲硫氨酸外，其它16种氨基酸的含量都高于对照，其中组氨酸、甘氨酸、苯丙氨酸、‘异亮氨酸、赖氨酸差异达显著水平，谷氨酞胺、脯氨酸差异达极显著水平，与氨基酸总量表现一致的变化趋势;取食KMDI后l天，除天门冬酸胺、谷氨酞胺和赖氨酸高于对照外，其它17种氨基酸均较对照低，但仅丝氨酸、苏氨酸和色氨酸与对照差异显著;取食K州Dl后3天，绝大多数氨基酸含量的变化与取食K加IDI后O天的趋势一致，除丙氨酸和色氨酸比对照稍低，其它18种氨基酸均高于对照，其中精氨酸、撷氨酸差异显著，丝氨酸差异达极显著。 取食MSB后0天，天门冬氨酸、天冬酞胺、谷氨酞胺和酪氨酸含量高于对照，丝氨酸、丙氨酸和精氨酸含量低于对照，但均差异不显著;取食MSB后1天，天门冬氨酸、谷氨酸、谷氨酞胺、组氨酸、甘氨酸、苏氨酸、丙氨酸、精氨酸、苯丙氨酸和脯氨酸的含量低于对照，与氨基酸总量变化趋势一致，但仅甘氨酸、苏氨酸、苯丙氨酸和脯氨酸差异显著或极显著;取食MSB后3天，仅天门冬氨酸和苯丙氨酸分别高于和低于对照，但差异都不显著。四、取食转基因抗虫水稻后二化螟幼虫血淋巴糖代谢和蛋白质浓度的变化 五龄二化螟幼虫取食KMDI后血淋巴总糖含量显著低于取食秀水11对照的，海藻糖酶活性显著高于对照，而海藻糖含量和葡萄糖含量与对照组相比都无显著差异;取食MSB后二化螟血淋巴总糖含量、海藻糖含量、葡萄糖含量以及海藻糖酶活性与取食明恢86的对照相比，均差异不显著。 取食KMDI后二化螟幼虫血淋巴蛋白质浓度显著低于取食秀水11的，其中处理后48h差异最大，但处理后不同时间差异不显著。二化螟幼虫取食MSB后血淋巴蛋白质浓度与取食明恢86的无明显差异;处理后不同时间差异也不显著。五、取食转基因抗虫水稻后二化螟幼虫中肠蛋白酶活性的变化 五龄二化螟取食KMDI后12h、24h、48h、60h和72h中肠蛋白质浓度与对照无显著差异;取食MSB后二化螟中?更多还原

【Abstract】 Two transgenic rice lines with high resistance to stem borer, [Bt rice (KMD1) and crylAb + sck rice (MSB)], three herbivores including two serious pests [stripped stem borer (SSB) Chilo suppressalis and brown planthopper (BPH) Nilaparvata lugens], one occasional pest [Mycalesis gotama] and three important natural enemies [Apanteles chilonis, Microvrlia horvathi, Pirata subpiraticus] were selected to study effects of transgenic rice lines on biological characteristics of the natural enemies through food chain and the relevant possible mechanisms in the laboratory. The main results were summarized as follows.1. Effects of transgenic Bt rice on survival and growth of SSB and biological characteristics of Apanteles chilonis via SSBThe corrected mortalities of 3rd, 4th and 5th instar larva of SSB continuously fed on KMD1 increased to 90.1%% 80.3% and 45.7% on the 4th day, respectively. The corrected mortalities of 3rd and 4th instar larva were 100% on the 6* and 7th day respectively, while that of 5th instar larva reached 96% on 8th day. Corrected mortality of SSB fed on control rice after fed on KMD1 for different times increased with feeding time on KMD1.The body weights of SSB fed on control rice after feeding on KMD1 for 36 h were very significantly lower on day 0 and day 3, and significantly lower on day 9, compared with those of the control.Parasitism rates of host larvae fed on KMD1 by Apanteles chilonis were significantly (3rd instar) or very significantly (4th and 5th instar) lower than those of the control host. Percentages of cocoon formation for 3rd and 4th instar hosts fed on KMD1 significantly declined. Pupa period of the parasitoid from the 3rd instar larva fed with KMD1 was significantly shorter, compared with the control. Longevity of the male wasps from the fifth instar larva fed on KMD1 was significantly shorter. Length of wasp cocoon of KMD1 treatment was significantly shorter. But no significant differences were detected in the development periods of egg to larva, cocoon mass size, wasp emergence rate and sex ratio between KMD1 and the control.2. Effects of transgenic cry 1 Ac + sck rice on survival and growth of SSB and biological characteristics of Apanteles chilonis via SSBMortality of SSB continuously feeding on MSB increased steadily, and was very significantly higher than that of MH 86 control on day 6 and afterwards. The weight of SSB continuously feeding on MSB was also significantly less than that of control on day 2 and afterwards.Mortalitis of SSB feeding on the control for 0 h 24 h 48 h 72 h after feeding on MSB for 36 h were all higher than those of the control, but the differences were not significantly. However, the weights of SSB were all very significantly less than those of the control.Mortalitis of SSB feeding on the control for 3 6 9 12 days after feeding on MSB for 36 h were higher than those of the control without significant differences; But the weights of the SSB were all lessthan those of the control, very significantly or significantly on day 3 and 6, respectively.Parasitism rates of host larvae fed on MSB were lower than those of the control, but only significantly for 4th instar. There were no significant differences for percentage of pupa formation between treatment and control. Development period of the parasitoid pupa from the 5th instar larva fed on MSB was significantly longer. Length of cocoon of MSB treatment was significantly shorter than the control. However, no obvious differences were detected in the development periods of egg to larva, cocoon mass size, wasp emergence rate, sex ratio, longevity and forewing length of wasp between KMD1 and the control.3. Amino acid content in the haemolymph of SSB after fed with transgenic riceAlthough there were no changes on the components of amino acids in the haemolymph of SSB after fed with two transgenic rice lines, there were some changes on relative contents of amino acids. Contents of total amino acids as well as His, Gly, Phe, He, Lys, Gin and Pro in haemolymph of SSB on更多还原