| 球孢白僵菌羧基转运蛋白基因RNA干扰的沉默效应 |
| Silencing effect of BbJEN1 gene of the entomogenous fungus Beauveria bassiana caused by RNA interference |
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| 中文关键词:马尾松毛虫，羧基转运蛋白，基因沉默，孢子发芽，毒力 |
| 英文关键词:Masson’s pine caterpillar, carboxylic transport protein, gene silence, conidium germination, virulence |
| 基金项目:“863”项目（No. 2006AA10A212）；教育部高等学校博士点专项科研基金（No. 200503644001） |
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| 中文摘要: |
| 羧基转运蛋白基因是近年来分离出的新基因，在虫生真菌中可能与穿透昆虫体壁时的能量代谢有关。构建了针对该基因的双链RNA干扰载体，采用芽生孢子转化法将载体质粒转入球孢白僵菌，并通过RT-PCR检测转化前后BbJEN1的表达情况。RT-PCR检测结果显示，球孢白僵菌转化子BbJEN1基因的表达水平显著下降，表明干扰载体具有明显的沉默效应。在两种培养基上，转化子与原始菌株的生长速度和产孢量差异均不显著，而分生孢子萌发则显著滞后于原始菌株。使用马尾松毛虫幼虫对转化子和原始菌株进行生物测定，原始菌株的半致死浓度、半致死剂量和半致死时间分别为2.79×106个孢子/mL、84.12个孢子/mm2和6.49d，而转化子的半致死浓度、半致死剂量和半致死时间则分别增加到1.27×107个孢子/mL、382.92个孢子/mm2和8.09d，毒力显著下降。由此表明BbJEN1基因与球孢白僵菌分生孢子发芽以及毒力均有关。 |
| 英文摘要: |
| The newly isolated carboxylic transport protein (CTP) gene of insect pathogenic fungi is implicated in energy metabolism during fungal penetration of insect integument. In the present study, a dsRNA interference vector was constructed, transformed into the blastospores of a Beauveria bassiana strain isolated from the Masson’s pine caterpillar, Dendrolimus punctatus, and the expression change of B. bassiana CTP gene BbJEN1 was examined by RT-PCR. The results indicated that the expression level of BbJEN1 was down-regulated substantially in the transformants. On two media, growth rate and the ability of sporulation between the transformant and the wild type strain were not significantly different, while the germination of the transformant was significantly reduced. Insect bioassay was conducted against the Masson’s pine caterpillar. The result showed that the median lethal concentration, median lethal dose and median lethal time of the wild type strain were 2.79×106conidia/mL, 84.12conidia/mm2 and 6.49 days, respectively, while those of the transformant were 1.27×107conidia/mL, 382.92conidia/mm2 and 8.09 days, respectively, indicating a significantly declined effect on virulence caused by RNA interference. The data of this study suggests that BbJEN1 gene is related to conidium germination and virulence. |
| 陈珊珊 郝蕾蕾 周娜 李增智. 球孢白僵菌羧基转运蛋白基因RNA干扰的沉默效应[J]. 菌物学报, 2010, 29(2): 273-278 |
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