农药对转Bt基因水稻生理生化及褐飞虱种群增长的影响
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摘要
随着转Bt基因水稻的研究的不断深入,所带来的环境安全性和食品安全性问题越来越受到学术界和社会公众的广泛关注。本文从农药胁迫的角度出发,研究农药对转Bt基因水稻的生理生化影响,以及农药和Bt毒蛋白对非靶标害虫褐飞虱的双重影响。结果如下：
1.农药对转Bt基因水稻和常规水稻受害级别的影响
本文研究了四种农药亚致死剂量对常规水稻和转Bt基因抗虫水稻受害级别的影响。结果表明农药处理并接虫后7天,水稻受害级别在农药浓度间有显著差异,但品种间差异不显著。三唑磷10ppm、20ppm、40ppm、80ppm处理后,MH63受害级别分别比对照增加了84.6%、61.5%、76.9%、76.9%；T2A-1受害级别分别比对照增加了26.7%、66.7%、40%、53.3%。康宽20ppm、40ppm、80ppm、160ppm处理后,MH63受害级别分别比对照增加了123%、92.3%、69.2%、92.3%；T2A-1受害级别分别比对照增加了93.3%、107%、66.7%、80%。接虫后14天,水稻受害级别随施药浓度和品种变化呈显著变化。除井冈霉素外,吡蚜酮、三唑磷和康宽处理后,农药浓度显著影响水稻受害级别,且在水稻品种间也有显著差异,另外农药浓度与水稻品种间存在显著的交互作用,例如,康宽40ppm处理的T2A-1受害级别高达8.2,比对照增加了64%。
2.农药对转Bt基因水稻和常规水稻生理生化的影响
转Bt基因作物的Bt毒蛋白对靶标害虫具有非常有效的控制作用,特别是鳞翅目害虫。然而,在Bt毒蛋白高选择压下,靶标害虫抗性的产生以及非靶标害虫种群数量增加使化学防治依然成为必要的手段。本研究的目的是比较农药对转Bt基因水稻和常规水稻生理生化的影响。结果显示,井冈霉素、吡蚜酮、三唑磷和康宽处理7天和14天后,农药浓度和水稻品种对水稻可溶性糖、草酸以及类黄酮含量均有显著影响。生理生化的动态变化趋势因农药类型和水稻品种而异,如井冈霉素、吡蚜酮、三唑磷和康宽处理14天后,MH63和T2A-1草酸含量随井冈霉素和康宽浓度的增加而逐渐下降,但吡蚜酮和三唑磷处理后,草酸含量均出现先降低后升高。井冈霉素、吡蚜酮和康宽处理后,T2A-1植株可溶性糖和类黄酮的含量均显著高于MH63,但草酸含量显著低于MH63。
3.农药对转Bt基因抗虫水稻和常规水稻上褐飞虱种群的影响
对非靶标害虫的影响是转Bt基因水稻安全性评价的重要内容。目前评价转Bt基因水稻对稻飞虱的影响主要侧重于水稻对稻飞虱的直接效应,转基因水稻上农药对褐飞虱的影响还没有被研究。本文参照庞雄飞(1992)方法构建褐飞虱种群生命表。结果表明,在无农药处理时(CK),转基因水稻T2A-1上褐飞虱产卵量显著高于MH63。农药处理后,两种水稻都出现了褐飞虱若虫羽化率和产卵量等参数不同程度的增加,其中褐飞虱产卵量随三唑磷浓度增加而增加,显著刺激了褐飞虱产卵,另外三唑磷10ppm处理的T2A-1褐飞虱下代初孵若虫数和种群趋势指数显著高于MH63。而井冈霉素和康宽处理的MH63褐飞虱卵孵化率显著高于T2A-1。上述结果表明,有些农药在转基因水稻上使用可能增加褐飞虱发生再猖獗的风险。
4.农药对转Bt基因水稻毒蛋白含量的影响及机理
对转Bt基因作物进行安全性评价,其中很重要的一个方面是评价Bt毒蛋白对生态环境的影响。而水稻Bt毒蛋白的表达和合成受诸多外在因素的影响。本研究主要是研究农药对水稻Bt毒蛋白的含量及影响机制进行探讨。结果显示,苗期井冈霉素、吡蚜酮、三唑磷和康宽处理,转Bt基因水稻毒蛋白含量均显著升高,特别是低浓度处理增加量最为显著。分蘖期和孕穗期,农药处理显著降低了水稻Bt毒蛋白含量,但对抽穗期水稻Bt毒蛋白无显著影响。为探讨农药影响Bt毒蛋白表达量的生理生化机制,我们研究了农药对Bt毒蛋白合成途径相关的酶类活性和生化物质的影响。结果发现,农药处理后GOT、GPT活性及游离氨基酸、可溶性蛋白含量均显著降低,回归分析表明,GOT、GPT活性及游离氨基酸、可溶性蛋白的变化与同时期的Bt毒蛋白含量呈正相关。由此我们认为,农药影响了转Bt基因水稻GOT、GPT活性及游离氨基酸、可溶性蛋白含量,从而干扰了Bt毒蛋白的合成。
Environmental and food safety derived from Transgenic Bt rice have caused extensive concern from academia and public because of its commercialized and large scale of plantation. The present paper selected pesticides as a stress factor, investigated the effects of pesticides on physiology and biochemistry of trans-Bt rice and the combined effects of pesticides and Bt toxicity protein on BPH. The main results are as follows:
1Effects of pesticides on injury level of conventional and transgenic Bt rice.
The present part examined effect of four pesticides on injury level of conventional (MH63) and transgenic Bt rice (T2A-1). Result indicated that injury level increased with an increase of pesticides concentration for both rice varieties at7days after insect release (7DAI), but no significant difference on the level was found between conventional and transgenic rice. The other three pesticides significantly influenced Injury indexes of MH63and T2A-1except for jingangmycin. For example, the injury index of MH63treated with10,20,40and80ppm triazophos increased by84.6,61.5,76.9, and6.9%and that of T2A-1increased by26.7,66.7,40and53.3%compared to untreated control, respectively; Injury indexes of MH63treated with20,40,80and160ppm chlorantraniliprole increased by123,92.3,69.2and92.3%and that of T2A-1increased by93.3,107,66.7and80%, respectively. For14DAI, pymetrozine, triazophos and chlorantraniliprole significantly influenced injury indexes of MH63and T2A-1, there was a significant difference between MH63and T2A-1. In addition, there were significant interactions between pesticide concentration and rice variety, with a maximum injury index (8.2scale) of T2A-1treated with40ppm chlorantraniliprole, increasing by64%compared to control.
2Effects of pesticides on biochemical substances of MH63and T2A-1
Bt toxicity protein has high control efficacy for target inset pests, especially for Lepidopteran since the release of transgenic crops. However, resistance of target pest and increase of non-target pests under high selective pressure still need to apply chemical pesticides to control non-target pests. Therefore, this study examined effects of pesticides on biochemical substances of conventional and transgenic rice and approached biochemical changes of T2A-1following pesticide applications. Results indicated that there were significant differences on soluble sugar, oxalic acid and flavonoid contents at7and14days after pesticide applications (7and14DAA) among four pesticides and between MH63and T2A-1. Changes of the substances varied with pesticide concentration and rice variety. For example, oxalic acid content of MH63and T2A-1decreased with increases of jinggangmycin and chlorantraniliprole concentrations at14DAA, but oxalic acid content of rice plants treated with pymetrozine and triazophos decreased and then increased. Soluble sugar and flavonoid contents of T2A-1treated with jinggangmycin, pymetrozine and chlorantraniliprole were significantly higher than those of MH63, but oxalic acid content was significantly lower than that of MH63.
3Effects of pesticides on Nilaparvata lugens Stal (Homptera:Delphacidae) feeding on MH63and T2A-1
Effect of transgenic Bt rice on non-target pests are an important aspect for the evaluation of safety. However, the evaluation of effect of transgenic Bt rice on planthoppers concentrates on direct effect of the rice to planthoppers. Effect of pesticides on planthoppers via the rice has not been understood until date. The present experiment constructed life table of laboratory population of N. lugens according to Pang (1992). Results showed that the number of laying-egg of females on T2A-1was significantly higher than that on MH63under without pesticide applications. Adult emergence rate and fecundity of adult females on both MH63and T2A-1treated with pesticides increased compared untreated control, of which triazophos significantly stimulated fecundity of N. lugens. Nymph numbers of next generation and population trend index on T2A-1treated with10ppm triazophos were significantly higher than those on MH63. Hatchability of eggs on MH63treated with jinggangmycin and chlorantraniliprole was significantly higher than that on T2A-1. These findings indicated that applications of some of pesticides on T2A-1will induce resurgence of N. lugens.
4Effects of pesticides on Bt toxicity protein content of transgenic rice and its mechanisms
Effect of Bt toxicity protein on ecological environment is another aspect of safety evaluation. Synthesis and expression level of Bt toxicity protein in plants are influenced by many factors. Results showed that toxicity protein content (TPC) at the seedling stage following the four pesticide applications significantly increased, especially with maximum increase for low concentration treatments. In contrast, TPC at the tillering and booting stages significantly decreased for pesticide applications. However, no impact of pesticides on TPC at the heading stage was found. In addition, the present experiment investigated effect of pesticides on the enzymes related to the synthesis of Bt toxicity protein. Results demonstrated that there was a positive correlation between and TPC and GOT or GPT activity and soluble protein. GOT and GPT activity, free ammonia acids and soluble protein of insecticide-treated T2A-1decreased significantly, indicating that pesticides interfered the synthesis of Bt toxicity protein through effects on GOT, GPT, free ammonia acids and soluble protein.
1.农药对转Bt基因水稻和常规水稻受害级别的影响
本文研究了四种农药亚致死剂量对常规水稻和转Bt基因抗虫水稻受害级别的影响。结果表明农药处理并接虫后7天,水稻受害级别在农药浓度间有显著差异,但品种间差异不显著。三唑磷10ppm、20ppm、40ppm、80ppm处理后,MH63受害级别分别比对照增加了84.6%、61.5%、76.9%、76.9%；T2A-1受害级别分别比对照增加了26.7%、66.7%、40%、53.3%。康宽20ppm、40ppm、80ppm、160ppm处理后,MH63受害级别分别比对照增加了123%、92.3%、69.2%、92.3%；T2A-1受害级别分别比对照增加了93.3%、107%、66.7%、80%。接虫后14天,水稻受害级别随施药浓度和品种变化呈显著变化。除井冈霉素外,吡蚜酮、三唑磷和康宽处理后,农药浓度显著影响水稻受害级别,且在水稻品种间也有显著差异,另外农药浓度与水稻品种间存在显著的交互作用,例如,康宽40ppm处理的T2A-1受害级别高达8.2,比对照增加了64%。
2.农药对转Bt基因水稻和常规水稻生理生化的影响
转Bt基因作物的Bt毒蛋白对靶标害虫具有非常有效的控制作用,特别是鳞翅目害虫。然而,在Bt毒蛋白高选择压下,靶标害虫抗性的产生以及非靶标害虫种群数量增加使化学防治依然成为必要的手段。本研究的目的是比较农药对转Bt基因水稻和常规水稻生理生化的影响。结果显示,井冈霉素、吡蚜酮、三唑磷和康宽处理7天和14天后,农药浓度和水稻品种对水稻可溶性糖、草酸以及类黄酮含量均有显著影响。生理生化的动态变化趋势因农药类型和水稻品种而异,如井冈霉素、吡蚜酮、三唑磷和康宽处理14天后,MH63和T2A-1草酸含量随井冈霉素和康宽浓度的增加而逐渐下降,但吡蚜酮和三唑磷处理后,草酸含量均出现先降低后升高。井冈霉素、吡蚜酮和康宽处理后,T2A-1植株可溶性糖和类黄酮的含量均显著高于MH63,但草酸含量显著低于MH63。
3.农药对转Bt基因抗虫水稻和常规水稻上褐飞虱种群的影响
对非靶标害虫的影响是转Bt基因水稻安全性评价的重要内容。目前评价转Bt基因水稻对稻飞虱的影响主要侧重于水稻对稻飞虱的直接效应,转基因水稻上农药对褐飞虱的影响还没有被研究。本文参照庞雄飞(1992)方法构建褐飞虱种群生命表。结果表明,在无农药处理时(CK),转基因水稻T2A-1上褐飞虱产卵量显著高于MH63。农药处理后,两种水稻都出现了褐飞虱若虫羽化率和产卵量等参数不同程度的增加,其中褐飞虱产卵量随三唑磷浓度增加而增加,显著刺激了褐飞虱产卵,另外三唑磷10ppm处理的T2A-1褐飞虱下代初孵若虫数和种群趋势指数显著高于MH63。而井冈霉素和康宽处理的MH63褐飞虱卵孵化率显著高于T2A-1。上述结果表明,有些农药在转基因水稻上使用可能增加褐飞虱发生再猖獗的风险。
4.农药对转Bt基因水稻毒蛋白含量的影响及机理
对转Bt基因作物进行安全性评价,其中很重要的一个方面是评价Bt毒蛋白对生态环境的影响。而水稻Bt毒蛋白的表达和合成受诸多外在因素的影响。本研究主要是研究农药对水稻Bt毒蛋白的含量及影响机制进行探讨。结果显示,苗期井冈霉素、吡蚜酮、三唑磷和康宽处理,转Bt基因水稻毒蛋白含量均显著升高,特别是低浓度处理增加量最为显著。分蘖期和孕穗期,农药处理显著降低了水稻Bt毒蛋白含量,但对抽穗期水稻Bt毒蛋白无显著影响。为探讨农药影响Bt毒蛋白表达量的生理生化机制,我们研究了农药对Bt毒蛋白合成途径相关的酶类活性和生化物质的影响。结果发现,农药处理后GOT、GPT活性及游离氨基酸、可溶性蛋白含量均显著降低,回归分析表明,GOT、GPT活性及游离氨基酸、可溶性蛋白的变化与同时期的Bt毒蛋白含量呈正相关。由此我们认为,农药影响了转Bt基因水稻GOT、GPT活性及游离氨基酸、可溶性蛋白含量,从而干扰了Bt毒蛋白的合成。
Environmental and food safety derived from Transgenic Bt rice have caused extensive concern from academia and public because of its commercialized and large scale of plantation. The present paper selected pesticides as a stress factor, investigated the effects of pesticides on physiology and biochemistry of trans-Bt rice and the combined effects of pesticides and Bt toxicity protein on BPH. The main results are as follows:
1Effects of pesticides on injury level of conventional and transgenic Bt rice.
The present part examined effect of four pesticides on injury level of conventional (MH63) and transgenic Bt rice (T2A-1). Result indicated that injury level increased with an increase of pesticides concentration for both rice varieties at7days after insect release (7DAI), but no significant difference on the level was found between conventional and transgenic rice. The other three pesticides significantly influenced Injury indexes of MH63and T2A-1except for jingangmycin. For example, the injury index of MH63treated with10,20,40and80ppm triazophos increased by84.6,61.5,76.9, and6.9%and that of T2A-1increased by26.7,66.7,40and53.3%compared to untreated control, respectively; Injury indexes of MH63treated with20,40,80and160ppm chlorantraniliprole increased by123,92.3,69.2and92.3%and that of T2A-1increased by93.3,107,66.7and80%, respectively. For14DAI, pymetrozine, triazophos and chlorantraniliprole significantly influenced injury indexes of MH63and T2A-1, there was a significant difference between MH63and T2A-1. In addition, there were significant interactions between pesticide concentration and rice variety, with a maximum injury index (8.2scale) of T2A-1treated with40ppm chlorantraniliprole, increasing by64%compared to control.
2Effects of pesticides on biochemical substances of MH63and T2A-1
Bt toxicity protein has high control efficacy for target inset pests, especially for Lepidopteran since the release of transgenic crops. However, resistance of target pest and increase of non-target pests under high selective pressure still need to apply chemical pesticides to control non-target pests. Therefore, this study examined effects of pesticides on biochemical substances of conventional and transgenic rice and approached biochemical changes of T2A-1following pesticide applications. Results indicated that there were significant differences on soluble sugar, oxalic acid and flavonoid contents at7and14days after pesticide applications (7and14DAA) among four pesticides and between MH63and T2A-1. Changes of the substances varied with pesticide concentration and rice variety. For example, oxalic acid content of MH63and T2A-1decreased with increases of jinggangmycin and chlorantraniliprole concentrations at14DAA, but oxalic acid content of rice plants treated with pymetrozine and triazophos decreased and then increased. Soluble sugar and flavonoid contents of T2A-1treated with jinggangmycin, pymetrozine and chlorantraniliprole were significantly higher than those of MH63, but oxalic acid content was significantly lower than that of MH63.
3Effects of pesticides on Nilaparvata lugens Stal (Homptera:Delphacidae) feeding on MH63and T2A-1
Effect of transgenic Bt rice on non-target pests are an important aspect for the evaluation of safety. However, the evaluation of effect of transgenic Bt rice on planthoppers concentrates on direct effect of the rice to planthoppers. Effect of pesticides on planthoppers via the rice has not been understood until date. The present experiment constructed life table of laboratory population of N. lugens according to Pang (1992). Results showed that the number of laying-egg of females on T2A-1was significantly higher than that on MH63under without pesticide applications. Adult emergence rate and fecundity of adult females on both MH63and T2A-1treated with pesticides increased compared untreated control, of which triazophos significantly stimulated fecundity of N. lugens. Nymph numbers of next generation and population trend index on T2A-1treated with10ppm triazophos were significantly higher than those on MH63. Hatchability of eggs on MH63treated with jinggangmycin and chlorantraniliprole was significantly higher than that on T2A-1. These findings indicated that applications of some of pesticides on T2A-1will induce resurgence of N. lugens.
4Effects of pesticides on Bt toxicity protein content of transgenic rice and its mechanisms
Effect of Bt toxicity protein on ecological environment is another aspect of safety evaluation. Synthesis and expression level of Bt toxicity protein in plants are influenced by many factors. Results showed that toxicity protein content (TPC) at the seedling stage following the four pesticide applications significantly increased, especially with maximum increase for low concentration treatments. In contrast, TPC at the tillering and booting stages significantly decreased for pesticide applications. However, no impact of pesticides on TPC at the heading stage was found. In addition, the present experiment investigated effect of pesticides on the enzymes related to the synthesis of Bt toxicity protein. Results demonstrated that there was a positive correlation between and TPC and GOT or GPT activity and soluble protein. GOT and GPT activity, free ammonia acids and soluble protein of insecticide-treated T2A-1decreased significantly, indicating that pesticides interfered the synthesis of Bt toxicity protein through effects on GOT, GPT, free ammonia acids and soluble protein.
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