›› 2005, Vol. 48 ›› Issue (2): 208-213.doi:

• 研究论文 • 上一篇    下一篇


陈  茂1,叶恭银1*,卢新民1,胡  萃1,彭于发2,舒庆尧3, Illimar ALTOSAAR4   

  1. 浙江大学应用昆虫学研究所/水稻生物学国家重点实验室
  • 出版日期:2006-06-06 发布日期:2005-11-20
  • 通讯作者: 叶恭银

Biotransfer and bioaccumulation of Cry1Ab insecticidal protein in rice plant-brown planthopper-wolf spider food chain

CHEN Mao1, YE Gong-Yin1*, LU Xin-Min1, HU Cui1, PENG Yu-Fa2, SHU Qing-Yao3, Illimar ALTOSAAR4   

  1. StateKey Laboratory of Rice Biology/Institute of Applied Entomology, Zhejiang University
  • Online:2006-06-06 Published:2005-11-20
  • Contact: YE Gong-Yin

摘要: 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。

关键词: Bt水稻, Cry1Ab杀虫蛋白, 褐飞虱, 拟水狼蛛, 食物链, 生物富集, 解毒

Abstract: The concentration of Cry1Ab insecticidal protein expressed in two homozygous transgenic Bacillus thuringiensis (Bt) rice lines, KMD1 and KMD2, were determined by enzyme immunosorbent assay (ELISA) with the PathoScreen kit for Bt cry1Ab/Ac protein (Agdia, USA). The results showed that the concentration of Cry1Ab expressed both in KMD1 and KMD2 at the heading and maturing stage were significantly lower than that at the seedling, tillering and booting stage. CrylAb protein could be transferred from   transgenic rice plants to its nontarget pest brown planthopper (BPH), Nilaparvata lugens (Stal), and from N. lugens to its natural enemy wolf spider (WS),Pirata subpiraticus (Boesenberg et Strand) in the food chain by preying. The quantitative cycle of Cry1Ab insecticidal protein in the food chain was analyzed using the kit. After being fed on KMD1 or KMD2 rice plants for 2 days, Cry1Ab protein could be detected in the N. lugens  body. However, after continuous feeding on Bt rice plants for 2, 4, 6, 8 and 10 days, the content of Cry1Ab remaining in the N. lugens body did not show the expected tendency of increase. Also, Cry1Ab protein could be detected in the P. subpiraticus body by preying on the N. lugens reared on KMD1 or KMD2 rice plants for 2, 4, 6, 8 or 10 days, while their contents were significantly higher than those of N. lugens  fed on KMD1 or KMD2 for the same time. Although the inoculation time was increased from 2 to 4, 6, 8 and 10 days, the level of Cry1Ab detected in P. subpiraticus did not show any increase.  Additionally, there were no significant differences among the predatory consumptions of P. subpiraticus preying on N. lugens reared on KMD1, KMD2 or on the non-transgenic parental rice line Xiushui11. No clear bioaccumulation pattern of Cry1Ab protein was observed in N. lugens and P. subpiraticus. More than 50 percent of Cry1Ab was degraded by the crude protease extract from the midgut of P. subpiraticus at 37℃ without light for 2 hours. However, as the exposure time increased from 2 to 4, 8, 12 and 24 hours, the detoxification function of the crude protease extract did not show a tendency to increase.

Key words: Bt rice, Cry1Ab insecticidal protein, Nilaparvata lugens;Pirata subpiraticus, food chain, bioaccumulation, detoxification