转基因抗虫耐除草剂玉米瑞丰125 Cry1Ab/Cry2Aj杀虫蛋白的时空表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2022-0339

摘要/Abstract

摘要：

玉米是我国最重要的粮食作物之一，虫害会造成严重的品质和产量损失，转Bacillus thuringiensis（Bt）基因抗虫玉米为玉米害虫的防治提供了新途径。转Bt基因抗虫玉米的抗虫性与外源杀虫蛋白表达量具有密切的关系，明确Bt杀虫蛋白在不同生育期和不同器官中的表达量，对害虫的综合防治和农业转基因生物安全管理具有重要意义。连续两年采用酶联免疫吸附法（ELISA）测定了9个地点种植的瑞丰125不同器官中Cry1Ab/Cry2Aj杀虫蛋白表达量，对比分析了玉米拔节期（V6-V8）叶片、抽雄期（VT）雄穗、吐丝期（R1）叶片和花丝、成熟期（R4）叶片和籽粒的Bt杀虫蛋白含量，结果表明Bt杀虫蛋白表达量在不同玉米器官中差异较大。2年9个地点的数据整体上呈现出叶片中Cry1Ab/Cry2Aj杀虫蛋白表达量较高，而籽粒中Cry1Ab/Cry2Aj杀虫蛋白表达量较低的规律。9个地点种植的瑞丰125的Cry1Ab/Cry2Aj杀虫蛋白表达量有所差异，但差异整体较小。

关键词: 转基因抗虫玉米, 瑞丰125, Cry1Ab/Cry2Aj杀虫蛋白, 时空表达

Abstract:

Maize is one of the most important food crops in China, but pests reduce the quality and yield of maize production. Genetically modified（GM）insect resistant maize expressing Bacillus thuringiensis（Bt）toxins are new tools for integrated pest management. The insecticidal effect of Bt-transgenic maize is closely related to the expression of exogenous insecticidal protein. Evaluating the expression of Bt insecticidal protein in different growth stages and tissues is of significance to the integrated pest management and biosafety management of agricultural GM organisms. In this paper, the expression of Cry1Ab/Cry2Aj insecticidal protein in different tissues of Ruifeng 125 planted in 9 places in two years was determined by ELISA. The Bt insecticidal protein expressions in the leaves at jointing stage（V6-V8）, tassels at tasseling stage（VT）, leaves and filaments at silking stage（R1）, leaves and grains at maturity stage（R4）were comparatively analyzed. It was found that the expressions of Bt insecticidal protein in different tissues of maize varied largely. The results showed that the expression of Cry1Ab/Cry2Aj insecticidal protein in the leaves was higher than that in the kernels. The expressions of Cry1Ab/Cry2Aj insecticidal protein of Ruifeng 125 from 9 different places were different, but the overall difference was slight.

Key words: genetically modified insect-resistant Bt maize, Ruifeng 125, Cry1Ab/Cry2Aj insecticidal protein, spatio-temporal expression

李东阳, 肖冰, 王晨尧, 杨现明, 梁晋刚, 吴孔明. 转基因抗虫耐除草剂玉米瑞丰125 Cry1Ab/Cry2Aj杀虫蛋白的时空表达分析[J]. 生物技术通报, 2023, 39(1): 31-39.

LI Dong-yang, XIAO Bing, WANG Chen-yao, YANG Xian-ming, LIANG Jin-gang, WU Kong-ming. Spatio-temporal Expression of Cry1Ab/Cry2Aj Insecticidal Protein in Genetically Modified Maize Ruifeng 125 with Stacked Insect and Herbicide Resistance Traits[J]. Biotechnology Bulletin, 2023, 39(1): 31-39.

图/表 4

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地点 Location	V6-V8叶片 V6-V8 leaf	VT雄穗 VT tassel	R1叶片 R1 leaf	R1花丝 R1 silk	R4叶片 R4 leaf	R4籽粒 R4 kernel
哈尔滨Harbin	5.659±1.090 a(ab)	1.645±0.374 b(b)	0.956±0.124 b(cd)	2.425±1.032 b(bcd)	2.001±0.229 b(e)	3.035±0.713 b(a)
长春Changchun	3.323±0.699 bc(cd)	2.110±0.336 cd(ab)	5.823±0.957 a(a)	1.608±0.239 d(cd)	4.038±0.119 b(bcde)	1.802±0.239 cd(bc)
沈阳Shenyang	4.084±0.189 bc(c)	2.731±0.157 c(a)	5.506±0.949 a(a)	0.800±0.164 d(d)	5.435±0.222 ab(abc)	1.213±0.348 d(bcd)
廊坊Langfang	2.099±0.249 cd(de)	2.874±0.682 bcd(a)	5.626±0.313 ab(a)	3.582±1.483 bc(bc)	6.415±1.394 a(a)	0.590±0.213 d(d)
烟台Yantai	4.660±0.351 a(bc)	1.926±0.088 b(ab)	5.182±0.500 a(a)	3.655±0.672 ab(bc)	3.226±1.480 ab(cde)	1.909±0.364 b(b)
新乡Xinxiang	1.716±0.269 c(e)	1.944±0.589 c(ab)	3.486±0.151 b(b)	3.184±0.321 b(bcd)	6.068±0.499 a(ab)	1.777±0.208 c(bc)
武汉Wuhan	3.995±0.510 a(c)	2.277±0.139 bc(ab)	2.729±0.316 b(b)	4.738±0.521 a(ab)	4.833±0.520 a(abc)	1.062±0.293 c(bcd)
贵阳Guiyang	6.577±0.165 a(a)	1.804±0.204 cd(ab)	2.510±0.416 c(bc)	6.716±0.933 a(a)	4.347±0.086 b(abcd)	0.674±0.175 d(d)
库尔勒Korla	1.137±0.250 b(e)	1.235±0.118 b(b)	0.841±0.186 b(d)	3.629±1.623 a(bc)	2.331±0.476 ab(de)	0.811±0.267 b(cd)

地点 Location	V6-V8叶片 V6-V8 leaf	VT雄穗 VT tassel	R1叶片 R1 leaf	R1花丝 R1 silk	R4叶片 R4 leaf	R4籽粒 R4 kernel
哈尔滨Harbin	5.659±1.090 a(ab)	1.645±0.374 b(b)	0.956±0.124 b(cd)	2.425±1.032 b(bcd)	2.001±0.229 b(e)	3.035±0.713 b(a)
长春Changchun	3.323±0.699 bc(cd)	2.110±0.336 cd(ab)	5.823±0.957 a(a)	1.608±0.239 d(cd)	4.038±0.119 b(bcde)	1.802±0.239 cd(bc)
沈阳Shenyang	4.084±0.189 bc(c)	2.731±0.157 c(a)	5.506±0.949 a(a)	0.800±0.164 d(d)	5.435±0.222 ab(abc)	1.213±0.348 d(bcd)
廊坊Langfang	2.099±0.249 cd(de)	2.874±0.682 bcd(a)	5.626±0.313 ab(a)	3.582±1.483 bc(bc)	6.415±1.394 a(a)	0.590±0.213 d(d)
烟台Yantai	4.660±0.351 a(bc)	1.926±0.088 b(ab)	5.182±0.500 a(a)	3.655±0.672 ab(bc)	3.226±1.480 ab(cde)	1.909±0.364 b(b)
新乡Xinxiang	1.716±0.269 c(e)	1.944±0.589 c(ab)	3.486±0.151 b(b)	3.184±0.321 b(bcd)	6.068±0.499 a(ab)	1.777±0.208 c(bc)
武汉Wuhan	3.995±0.510 a(c)	2.277±0.139 bc(ab)	2.729±0.316 b(b)	4.738±0.521 a(ab)	4.833±0.520 a(abc)	1.062±0.293 c(bcd)
贵阳Guiyang	6.577±0.165 a(a)	1.804±0.204 cd(ab)	2.510±0.416 c(bc)	6.716±0.933 a(a)	4.347±0.086 b(abcd)	0.674±0.175 d(d)
库尔勒Korla	1.137±0.250 b(e)	1.235±0.118 b(b)	0.841±0.186 b(d)	3.629±1.623 a(bc)	2.331±0.476 ab(de)	0.811±0.267 b(cd)

地点 Location	V6-V8叶片 V6-V8 leaf	VT雄穗 VT tassel	R1叶片 R1 leaf	R1花丝 R1 silk	R4叶片 R4 leaf	R4籽粒 R4 kernel
哈尔滨Harbin	4.222±0.540 ab(c)	1.888±0.046 b(ab)	3.182±0.297 b(b)	2.968±0.129 b(bc)	6.172±1.843 a(ab)	1.714±0.501 b(a)
长春Changchun	4.226±0.358 a(c)	1.626±0.243 d(b)	2.876±0.659 bc(b)	2.041±0.220 cd(d)	3.443±0.545 ab(c)	0.326±0.133 e(b)
沈阳Shenyang	11.133±0.420 a(a)	1.970±0.499 d(ab)	7.164±0.498 b(a)	1.575±0.163 d(de)	3.815±0.550 c(bc)	1.974±0.589 d(a)
廊坊Langfang	2.245±0.887 c(d)	1.404±0.285 c(b)	4.411±0.586 b(b)	2.315±0.224 c(cd)	6.310±0.657 a(a)	1.025±0.075 c(ab)
烟台Yantai	2.494±0.351 bc(d)	1.808±0.131 c(ab)	4.495±1.580 ab(b)	2.285±0.349 bc(cd)	4.883±0.667 a(abc)	1.488±0.450 c(a)
新乡Xinxiang	6.319±0.317 a(b)	1.978±0.184 cd(ab)	3.772±0.142 b(b)	3.212±0.473 bc(b)	6.330±0.916 a(a)	1.311±0.484 d(ab)
武汉Wuhan	3.045±0.620 b(cd)	2.112±0.359 bc(ab)	4.802±0.126 a(b)	1.661±0.251 cd(d)	2.583±0.309 bc(c)	1.040±0.094 d(ab)
贵阳Guiyang	2.483±0.045 b(d)	2.423±0.101 b(a)	4.760±1.132 a(b)	0.762±0.269 c(e)	4.858±0.456 a(abc)	1.034±0.113 bc(ab)
库尔勒Korla	2.538±0.341 b(d)	1.804±0.050 bc(ab)	2.802±0.522 b(b)	4.486±0.285 a(a)	4.214±0.425 a(abc)	0.983±0.277 c(ab)

地点 Location	V6-V8叶片 V6-V8 leaf	VT雄穗 VT tassel	R1叶片 R1 leaf	R1花丝 R1 silk	R4叶片 R4 leaf	R4籽粒 R4 kernel
哈尔滨Harbin	4.222±0.540 ab(c)	1.888±0.046 b(ab)	3.182±0.297 b(b)	2.968±0.129 b(bc)	6.172±1.843 a(ab)	1.714±0.501 b(a)
长春Changchun	4.226±0.358 a(c)	1.626±0.243 d(b)	2.876±0.659 bc(b)	2.041±0.220 cd(d)	3.443±0.545 ab(c)	0.326±0.133 e(b)
沈阳Shenyang	11.133±0.420 a(a)	1.970±0.499 d(ab)	7.164±0.498 b(a)	1.575±0.163 d(de)	3.815±0.550 c(bc)	1.974±0.589 d(a)
廊坊Langfang	2.245±0.887 c(d)	1.404±0.285 c(b)	4.411±0.586 b(b)	2.315±0.224 c(cd)	6.310±0.657 a(a)	1.025±0.075 c(ab)
烟台Yantai	2.494±0.351 bc(d)	1.808±0.131 c(ab)	4.495±1.580 ab(b)	2.285±0.349 bc(cd)	4.883±0.667 a(abc)	1.488±0.450 c(a)
新乡Xinxiang	6.319±0.317 a(b)	1.978±0.184 cd(ab)	3.772±0.142 b(b)	3.212±0.473 bc(b)	6.330±0.916 a(a)	1.311±0.484 d(ab)
武汉Wuhan	3.045±0.620 b(cd)	2.112±0.359 bc(ab)	4.802±0.126 a(b)	1.661±0.251 cd(d)	2.583±0.309 bc(c)	1.040±0.094 d(ab)
贵阳Guiyang	2.483±0.045 b(d)	2.423±0.101 b(a)	4.760±1.132 a(b)	0.762±0.269 c(e)	4.858±0.456 a(abc)	1.034±0.113 bc(ab)
库尔勒Korla	2.538±0.341 b(d)	1.804±0.050 bc(ab)	2.802±0.522 b(b)	4.486±0.285 a(a)	4.214±0.425 a(abc)	0.983±0.277 c(ab)