Studies On The Introduction Of Bt Gene Into Kenaf And The Inheritance Genetic Stability

In order to promote the development of kenaf breeding in insect-resistant transgene, six types of state-level spreading kenaf varieties with high yield high quality and disease-resistant been injected Bt gene by Pollen Tube Pathway , that is stigma addition and ovary injection. The insect-resistant effect were detected in Bt transgenic variety of Fuhong 952 for 4 generations by field investingating and insect feeding ,as well as by PCR and ISSR marker-assisted detection, Southern molecular hybridization confirmation, the result obtained showed that:1 The preparation and purification bt plasmid:preparing the plasmid which contains Bt gene, OD_260/280 between 1.9-2.0 was available after purified. 2 The genetic transfer efficiency :deal with 470 flowers through stigma addition and ovary injection, the progeny setting seed rate were 68.8% and 51.7% respectively, inherence less insect eating plants were 7.2% and 24.5% respectively, and the total transfer efficiency of the two procedures was 0.48% by PCR detecting, and obtained the varieties of fuhong 952 which has insect-resistant character. 3 Identification of insect feeding on the culture plate: put the insect-resistant and nontransgene kenaf leaves as comparation, feed the insect on the culture plate and the insect-resistant has insect-resistant character.4 PCR molecular detection of target gene: By detecting different generations kenaf leaves with PCR molecular detection, the result showed that they are all have the target banes5 ISSR molecular marker detection target gene: By detecting the insect-resistant kenaf and nontransgenic kenaf with ISSR molecular marker detection for the first time,the result showed that they have polymorphisms. The result showed that ISSR molecular marker detection can be used as a way of insect-resistant detection. 6 Southern molecular hybridization detection: Detecting four generations of transgenic kenaf with digoxin Southern molecular hybridization ,and there are three target gene copies in the four generations, which were 2500bp 1500bp 680bp copies. 7.. Breeding of the varieties fuhong 952 with insect-resistant gene: obtained three well-resistant insect strains with the detection of nature from 97 different individuals, and bred the new kenaf varieties with insect-resistant and high-production of...