Font Size: a A A

Bioassay Of Vegetative Insecticidal Proteins Against The Forests Insects And The Construction Of The Plant Expression Vectors

Posted on:2009-08-11Degree:MasterType:Thesis
Country:ChinaCandidate:L L LiuFull Text:PDF
GTID:2143360242987458Subject:Tree genetics and breeding
Abstract/Summary:PDF Full Text Request
Bacillus thuringiensis (BT) have been developed as one of the most successful animalcule in agriculture to control insects. The thesis is to investigate bioassay of vip3 A against the forest insects as well as plants espression vector construction and genetictransformation.1 .Research on bioassay of vip3A against the forest insectsInsecticidal Crystal Proteins, ICPs are toxic to agriculture importance insects.TheVip3A are newly discovered proteins which differ from ICPs in function mechanism. It is not repcrted thatVip3A proteins are toxic against Hyphantria cunea and Clostera anachoreta.Recombinant vectors of pPV113 and pPV116 were constructed separately by inserting Vip3A-LS1 gene and Vip3A-LS8 gene into pET21-b vector. 88kD protein of Vip3A was acaquired by SDS-PAGE. The Bioassay showed that the proteins showed a high toxicity against the 1st larvae of Hyphantria cunea and Clostera anachoreta. the corrected mortality rates against that two insects larvar were (53.02±0.43)% and (68.00±0.60)% respectively with 80μg/m Vip3A-LS1 protein. The weighteinhibitory rate against H. cunea 1st larvae was 80.60%when it is 80μg/mL Vip3A-LS8protein. the corrected mortality rates against that two pests were (52.12±0.53)% and (70.0±0.53)% respectively. The weight inhibitory rate against H. cunea 1st larvae was 78.90%.It is firstly discovered that Vip3 A are highly toxic against forests insects.2. Research on plants espression vector construction and genetictransformation.As plant biotechnology grows into the mature stage and large scale application of insect resistant plants expressing only kind of BT crystal protein would probably force other target insects to develop tolerance against this type of plants just like the case happened in the large scale application of Bt sprays in the field ,a strategy of using transgenic plants expressing more than one insecticidal proteins with different mechanisms of action against the target insects has been proposed to avoid or delay the development of this kind of tolerance.The Vip3A gene in a size of 2.37kb amplified from the recombinant plasmids pPV113 and pPV116 by PCR was cloned into expression vector pBI121. The expression vector p1302PV113 and p1302PV116 was constructed by inserting Vip3A.By Agrobacterium tumefaciens-mediated transformation ,The two expression vectors were respectively transferred into two kinds one is male Populus tomentosa and the other conctains BTgene. The transformed buds were selected on the medium containing kan and rootage . The PCR analysis and GFP histochemical assays showed that Vip3A gene had integrated into the genome of the populus tomentosa plants.
Keywords/Search Tags:Bacillus thuringiensis, vegetative insecticidal proteins, Hyphantria cunea, Clostera anachoreta, plants expression vector, male Populus tomentosa, transformation
PDF Full Text Request
Related items