Studies On The Biosafety Of Transgenic Insecticidal Cotton (Gossypium Hirsutum L.) To Itself And Soil Ecology

Cotton cultivar SGK321, SHIYUAN321, 99B and ZHONG29 as materials, ELISA as the main method, this paper studied on how exogenous genes affect the resistance of the Insect-Resistant cotton and the soil ecosystem. Results are as follows:1 CpTI was purified by G-75 column and affinity chromatography from 901 QINGPI cowpea which was selected from several varieties by trypsin inhibitor activity. The polyclonal antibody against CpTI was prepared and a enzyme immunoassay was established. CpTI content in different parts of the transgenic cotton were determined. The results show that the expression levels of the exogenous genes are various in different organs. The results showed that the expression of different organs of the same cultivar was different.2 Insecticidal crystal protein were checked by ELISA. The content in rhizospere system is higher than in periphery,and transgenic cotton than general cotton. In prophase Insecticidal crystal protein near cotton is more than distant soil to the same cultivar, but the same during anaphase. CpTI is fewer, but positive appearently. Bt and CpTI both arrive at perk respectively when plant expressed Insecticidal crystal protein most.3 Content of Bt-toxin and CpTI were checked with the method of ELISA during all the growth phase. Conclusion shows the both toxins accumulate in rhizospere of Insecticidal cotton. But CpTI is lower than Bt-toxin. Treated with plant growth regulator DPC, the exudation of Bt and CpTI increased.4 Compared with regular cotton, content of gossypol and the activity of P -1, 3-glucanase and chitinases is high in the late growth phase; but condensed tannin and proline is low. The latter can be the cause of weakness of insecticidal-cotton's resistence, which can induced the former. Treated with DPC, all the substances increased.5 The sum of four kinds of bacteria in rhizospere of Insecticidal cotton is less than regular cotton at the whole growth phase. Next year before planting cotton the four bacteria restore to the level of regular cotton. No discipline were found after treated with the plant growth regulator DPC.6 Bt insecticidal protein in different Organs of transgenic Insect-Resistant Cotton was localized by immuno-histochemical method. The results showed that organs of Transgenic Insect-Resistant Cotton was obviously positive, the ordinary cotton was negtive. From these pictures, the expression of Bt Insecticidal Protein in different insect-resistent varieties, different organs of the same variety and different tissues of the same organs of transgenic insect-resistant Cotton is critically different, especially the young boll. Because the sample consists of several tissues, We consider the quantification of Bt Insecticidal Protein by ELITSA or bioassay is not enough.