Application of chilli DN (dominant negative) mutant to bacterial wilt resistance gene engineering of tobacco

A technology of genetic engineering and mutants, applied in the fields of genetic engineering, application, plant gene improvement, etc., can solve the problems of ROP functional redundancy, ROP with few phenotypic changes, etc., and achieve the effect of improving bacterial wilt resistance

Inactive Publication Date: 2012-11-14
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

[0005]Since the discovery of ROP in plants, more and more studies have found that ROP plays a key role in various signaling pathways in plants, and each member may act on different Several different pathways may be integrated into the same ROP (Jones et al., 2005; Yalovsky et al., 2008), and there is also functional redundancy of ROP in plants, resulting in the lack of a certain ROP. There are few obvious phenotypic changes, so gene function knockout alone is rarely used in the study of ROP
There is no literature report on the use of DN mutants to study plant resistance to bacterial wilt

Method used

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  • Application of chilli DN (dominant negative) mutant to bacterial wilt resistance gene engineering of tobacco
  • Application of chilli DN (dominant negative) mutant to bacterial wilt resistance gene engineering of tobacco
  • Application of chilli DN (dominant negative) mutant to bacterial wilt resistance gene engineering of tobacco

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Embodiment Construction

[0020] 1 Vector construction process

[0021] 1.1 Materials

[0022] The pepper variety is the local pepper variety 120 in Lianhua County, Jiangxi Province # , preserved by the laboratory of the College of Life Sciences, Fujian Agriculture and Forestry University; the flue-cured tobacco variety K326 was donated by the Virus Institute of Fujian Agriculture and Forestry University; the cDNA library of pepper leaves treated with UV-B (UV-B ultraviolet treatment: put the plants under a UV lamp (UV -B) 2h, 6h, 12h, 24h and 48h were irradiated at the lower 25cm, and all plants treated in different treatment periods were quick-frozen with liquid nitrogen and then stored in a -80°C refrigerator) for the laboratory of the School of Life Sciences, Fujian Agriculture and Forestry University Constructed (operated according to the instructions on the Invitrogen kit), the titer was as high as 1×10 7 ; CloneMiner TM Both cDNA Library Construction Kit and Gateway Vector Construction Kit w...

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Abstract

The invention provides application of a chilli DN (dominant negative) mutant to the bacterial wilt gene engineering of tobacco. The chilli DN mutant is a DN-CaROP1 mutant. A gene of the mutant at least comprises a nucleotide sequence shown as SEQ ID No.1. A construction method comprises a step of fusing the chilli DN-CaROP1 mutant with a core sequence of a CaMV35S promoter to construct a DN-CaROP1 over-expression carrier. Compared with wild tobacco, by over-expression of the chilli DN-CaROP1 mutant in tobacco, the bacterial wilt resistance capability of the transgenic tobacco is obviously improved. The mutant has very important application value in the bacterial wilt gene engineering of plants.

Description

technical field [0001] The present invention relates to the construction of a capsicum DN-CaROP1 mutant and its overexpression vector and its application in genetic engineering, and more specifically relates to the application of the capsicum gene in the genetic engineering of tobacco bacterial wilt resistance in Solanaceae plants. The invention belongs to the technical field of plant genetic engineering. Background technique [0002] Plant growth, development and environmental response are realized through complex physiological and biochemical processes under the interaction of its specific genetic system and environmental conditions, and are always subject to relatively strict regulation so that it can produce enough photosynthetic products and reasonably The distribution and use of these photosynthetic products ultimately enable plants to obtain the greatest chance of survival and reproduction in a specific environment. Signal transduction composed of calcium messenger s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82
Inventor 何水林邱爱连蔡汉阳刘林林官德义刘志钦陈彦生牟少亮
Owner FUJIAN AGRI & FORESTRY UNIV
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