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Transgenic Indica Rice Harboring Two Insect-resistance Genes Obtained by Agrobacterium Tumefaciens-mediated Transformation and the Preliminary Research of Their Descendants

Author: DiHongLi
Tutor: XieHuaAn；WangFeng
School: Fujian Agriculture and Forestry University
Course: Crop Genetics and Breeding
Keywords: Indica Transgenic Bivalent genes Insect-resistant
CLC: S511.21
Type: Master's thesis
Year: 2002
Downloads: 111
Quote: 1
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Abstract


Transgenic Bt plants in recent years has been widely used in the production. However, with the large-scale cultivation of insect-resistant transgenic plants, the insect-resistant spectrum is relatively narrow and pests easy resistance and other issues more and more attention. Therefore, in order to turn bivalent or polyvalent insect-resistant GM plants as representative of the second generation of insect-resistant transgenic plants to become one of the hot spots in the study of plant genetic engineering. Rice is the most important food crop in the world, with the increase in population and the reduction of arable land, its significance to further improve rice yields. The pest is one of the main limiting rice yield, the use of transgenic technology to cultivate insect-resistant rice rice pest problems can be an effective solution. In this study, carried out the following work: 1 to insecticidal spectrum obtained with a wider and longer application period for the purpose of transgenic insect-resistant rice, two insecticidal genes that sck gene and Bt gene and marker gene tide neomycin gene (hpt) build-Hyg the pCUBAC in the same expression vector. Expression vector into Agrobacterium tumefaciens LBA4404 by electroporation. Agrobacterium-mediated transformation excellent indica Minghui 86 immature embryos induced embryogenic callus after six batch conversion, hygromycin resistant callus 70 conversion rate of 3.22% . 53 clones differentiate into seedlings, transplant survival of 45 clones (a total of 162 regenerated plants). Detected by PCR and the PCR-Southern hybridization analysis showed that in the regeneration of plants, the 137 transformed plants having two insecticidal genes at the same time, 16 contains only sck gene 9 as false positive. 2, the detection of the protein expression of Bt transgenic rice obtained results show that transgenic plants expressing Bt protein content of 0.05% -0.12% of the total soluble protein in rice leaves. T 1 -generation T 2 progeny PCR assay and ELISA found some transgenic plants genome the Bt gene failed to express the Bt protein transgenic silencing. Field insect resistance of transgenic plants obtained the survey results show that most of the transgenic plants were resistant lepidopteran pests such as stem borer. T 2 generation 13 homozygous lines, provides excellent material for parental breeding for insect-resistant transgenic rice. In addition, the separation ratio measurement erratic phenomenon were discussed.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Rice > Meters of sticky points > Indica
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