Study On The Agrobacterium Tumefaciens-mediated Transformation Of Bt Gene Into Sea Island Cotton(gossypium Barbadense L.)

In this study, the hypocotyls of six Sea Island cottons was used as the explant to study the regeneration system, i. e. DJ-1(373)、Tianchang 2(376)、5917(380)、Achang-599(381)、Ta 07-152(383)、S0717(408).The embryogenic callus of XinHai 14 and XinHai 33 was used to study the agrobacterium tumefaciens-mediated transformation system of Bt gene into Sea Island cottons, PCR and GUS histochemical examination was used to the test graft-transgenic plants, the preliminary study suggests exogenous Bt gene was already transformed into the genome of the two species of sea island cotton. The followings are the main results of this study:Calli were induced successfully on MSB medium applied with two different plant growth regulator(PGR) combinations selected with the method of double factors randomized block design. The best PGRs combinations to induce callus for 380、383、381 was 0.1 mg/L 2,4-D and 0.1 mg/L KT, while that to induce callus for 373、376、408 was 0.3 mg/L NAA and 0.1 mg/L KT. Somatic embryogenesis were observed on MS5 medium with 0.1 mg/L KT and 0.05 mg/L IBA. The culture of deformity seedlings can get more somatic embryogenesis and cotyledon embryos, the regeneration plant of the 6 varieties can be induced in six to eight months, so that a stable and efficient regeneration system that without genotype limits could be developed to produce new cotton varieties for cultivation in Xinjiang and lay the foundation to broaden the sea island cotton regeneration genes and transgenic research.The embryogenic callus of XinHai 14 and Xin Hai 33 as the receptors materials to optimize agrobacterium tumefaciens-mediated transformation system of Bt gene genetic transformation system. The conversion rate is improved if embryogenic calli was pre-cultured for ten days, and then infect for 15 minutes and co-culture for 24 hours,then was transferred to the selection medium with 50mg/L Kan and 500 mg/L cef. The incidence of somatic embryo is proved when 3.8 g/L KNO3 and 0.55 g/L NH4NO3 was added into the differentiation medium. 1/2MS in seedlings medium is beneficial to the development of normal cotyledons embryo.PCR and GUS histochemical detection was used to the test the ten graft-transgenic plants, the preliminary study suggests exogenous Bt gene was already transformed into the genome of the two species of sea island cotton.