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A Sensitive Bioassay for the Detection of Cry1A Toxin Expression in Transgenic Cotton

Cry1Ac公司 生物测定 生物 龄期 苏云金杆菌 幼虫 毒素 毒理 棉属 转基因 转基因作物 植物 基因 微生物学 生态学 生物化学 遗传学 细菌
作者
K. R. Kranthi,S. Kranthi
出处
期刊:Biocontrol Science and Technology [Taylor & Francis]
卷期号:10 (5): 669-675 被引量:7
标识
DOI:10.1080/095831500750016460
摘要

In the process of development of insect resistant transgenic plants and also to evaluate the consistency in expression of the toxin under greenhouse and field conditions, immunological and bioassays are commonly used. The assay being described in this report, is based on the high levels of sensitivity of a cotton leaf feeding insect, the semilooper, Anomis flava (Fabricius) to Cry toxins (Cry1Aa, Cry1Ab and Cry1Ac). The assay is sensitive, quick and reproducible. Cry1Ac was the most toxic followed by Cry1Ab and Cry1Aa. LC 50 s of the three toxins on first instar larvae ranged from 0.79-6.08 ng cm -2 of leaf. LC 50 s of Cry1Ac for the fourth instar larvae ranged from 12.91-21.14 ng cm -2 while LC 50 s for Cry1Aa and Cry1Ab were in the range 53.0-138 ng cm -2 . The fiducial limits (at 95% probability) of the probit assay data indicated that there was no difference in response between the three different populations to each of the three toxins. The data from all assays were pooled for each of the three toxins separately and subjected to regression analysis to obtain a cumulative log dose response for first and fourth instar larvae. These can be used as standard curves to quantify toxin expression in plants based on mortality response of either first or fourth instar A. flava larvae. Apart from being used to detect expression in putative Bt cotton transgenic plants, the assay can also be used to follow the activity of Cry toxins in transgenic cotton plants in the field during the growing season.

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