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Bollgard-II抗虫棉基因表达稳定性评估

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Bollgard-II抗虫棉基因表达稳定性评估

发布日期:2018-12-26 作者: 点击:

遗传稳定性是评估转基因作物开发价值和安全性的重要指标。遗传稳定性的评价通常是从目的基因整合、目的基因表达定量和目标性状的稳定性3个维度展开。其中,目的基因表达的稳定性评估需要考虑受体的背景基因型、转化体种植时间、种植地点、发育时期、组织器官等各方面因素对基因表达的影响。

孟山都公司开发的第二代转基因抗虫棉Bollgard-II表达Cry1Ac和Cry2Ab2两个Bt蛋白。由于Cry1Ac和Cry2Ab的氨基酸同源性很低且结合在昆虫中肠的不同位点,从而使得昆虫对它们产生交互抗性的概率很低,这就有助于延缓害虫抗药性的产生,延长转基因作物的市场寿命。Bollgard-II抗虫棉2006年起在印度开始种植,并已导入很多当地的棉花杂交种的亲本种质资源中。

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近期,研究者将印度北部、中部和南部棉花种植区12个种植季的2000多份杂交棉中,目的基因在新叶、开花前的棉蕾、棉铃表皮3个不同组织中的蛋白表达量进行了检测,并利用线性混合效应模型,评估影响蛋白表达量的因素,为综合评估Bollgard-II抗虫棉的基因表达调控稳定性提供了丰富的数据。结果表明,Cry1Ac蛋白在叶片和棉铃中的表达受基因型、种植季、发育时期共同影响,而在棉蕾中的表达主要受种植季的影响;Cry2Ab在3个组织中的表达都受到种植季的影响。两个蛋白在棉蕾中的表达变异性很强烈。另外,Bt蛋白在234份陆地棉和海岛棉的杂交种中虽然表达量有所不同,但是却呈现出了相似的表达模式。


Transgenic Research, 30 October 2018

Stability of expression of Cry1Ac and Cry2Ab2 proteins in Bollgard-II hybrids at different stages of crop growth in different genotypes across cropping seasons and multiple geographies


Author

Anik Luke Dhanaraj*, Alan Ray Willse,S. P. Kamath

*: Monsanto Research Centre, India


Abstract

Bollgard-II cotton expressing Cry1Ac and Cry2Ab2 insecticidal proteins has been commercially cultivated in India since 2006 to control bollworms. These genes were introgressed into parental germplasm of numerous hybrids. Therefore, it is imperative that these insecticidal proteins are expressed in sufficient quantities in different tissues, throughout the season irrespective of genetic background or environmental conditions for effective performance. Here, we document results of a comprehensive study on pattern of expression of Bt proteins across different stages of crop growth in > 2000 cotton hybrids (Gossypium hirsutum), across 12 cropping seasons tested in the Northern, Southern or Central zones in India, interminal leaf, pre-candle square and boll epicarp tissues. Statistical analysis of variability using Linear mixed effect model was used to estimate factors contributing to variability in expression of Bt proteins. For Cry1Ac, variability was maximally contributed by genotype × season × plant growth stage effect in terminal leaves and boll epicarp, while season effect drove variability in pre-candle square. In Cry2Ab2, season effect drove variability in three tissue types. Pre-candle square tissue had most variability in expression of both proteins followed by terminal leaf and boll epicarp. Further, expression of Bt proteins in 234 G. hirsutum × G. barbadense hybrids showed similar expression patterns as intra specific hybrids though there was a significant difference in expression levels. Cry2Ab2 was expressed in significantly higher amounts when genes were in homozygous state. Bt proteins were also found to be expressed in varied amounts in different tissues and were expressed even when hybrids were grown at sub-optimal temperatures.


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关键词:基因表达定量,基因表达调控,基因表达

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