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Salt resistance gene StNHX1 of wild eggplant and herbicide resistance plant expression vector thereof

A plant expression vector, wild eggplant technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., to achieve the effect of improving salt tolerance and broad application prospects

Inactive Publication Date: 2011-02-16
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of improving the salt tolerance of crops and provide a new wild eggplant salt tolerance gene StNHX1
[0005] Another object of the present invention is to provide the herbicide-resistant plant expression vector and construction method of the salt-tolerant gene StNHX1 in order to simultaneously solve the problems of crop salt tolerance and herbicide resistance

Method used

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  • Salt resistance gene StNHX1 of wild eggplant and herbicide resistance plant expression vector thereof
  • Salt resistance gene StNHX1 of wild eggplant and herbicide resistance plant expression vector thereof
  • Salt resistance gene StNHX1 of wild eggplant and herbicide resistance plant expression vector thereof

Examples

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Effect test

Embodiment 1

[0039] Example 1 Cloning of wild eggplant salt tolerance gene StNHX1

[0040] Wild eggplant 'Torvum Vigor' (Solanum torvum Swartz; Japanese rootstock variety, purchased from Japan Takii Seedling Co., Ltd.) was selected as the test material. When the seedlings grew to eight true leaves, 100 mmol L -1 After NaCl stress treatment for 3 days, young leaves were taken, and total RNA was extracted according to the instructions of the Total RNA Extraction Kit (TaKaRa), and 2 μg of total RNA was reverse-transcribed into cDNA (ReverTra Ace-a- TM cDNA synthesis kit, Toyobo (Shanghai) Biotechnology Co., Ltd.); according to the sequence information of tomato NHX1 published on NCBI (Access NO.: AJ306630.1; Venema K, Belver A, Marin-Manzano MC, Rodriguez-Rosales MP ,Donaire JP.A novel intracellular K + / H + Antiporter related to Na + / H + Antiporters are important for K + ionhomeostasis in plants.J Biol Chem.2003,278:22453-22459), with Oligo6.0 primer design software (Germany Merck comp...

Embodiment 2

[0044] 1) The cloning of the wild eggplant salt tolerance gene StNHX1 is the same as in Example 1.

[0045] 2) Construction of intermediate vector pCAMBIA3301-121

[0046] pBI121 (Promega, Promega (Beijing) Biotechnology Co., Ltd.) and pCAMBIA3301 (CAMBIA, Australia) were digested with Hind III / EcoR I respectively, and the small fragment (3005bp) in pBI121 and the large fragment (11251bp) in pCAMBIA3301 were recovered ), according to the ratio of 4:1 with T 4 DNA ligase (TaKaRa) ligation, named pCAMBIA3301-121, the ligation product was transformed into DH5α competent cells, and the plasmid was extracted for Hind III / EcoR I double digestion verification. The specific steps are as follows:

[0047] ①Take 10 μL each of pBI121 and pCAMBIA3301 plasmids, and digest them with Hind III / EcoR I respectively. Reaction system for enzyme digestion (50 μL): ddH 2 O 31.5 μL, 10×Buffer E 5 μL, BSA 0.5 μL, pBI121 or pCAMBIA3301 plasma 10 μL, Hind III (Promega) 1.5 μL, EcoR I (Promega) 1.5 μL...

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Abstract

The invention belongs to the field of molecular biology, disclosing a salt resistance gene StNHX1 of a wild eggplant and an herbicide resistance plant expression vector thereof. The sequence of the salt resistance gene StNHX1 of the wild eggplant is SEQ ID NO.1. The herbicide resistance plant expression vector of the gene is prepared by inserting the StNHX1 genes among BamHI / Sac I enzyme cutting sites of an intermediate vector pCAMBIA3301-121. The plant expression vector promotes crops to have the characteristic of herbicide resistance and enhance salt resistance properties after transforming the crops, and has important theoretical and practical significance on enhancing the technical level of the molecular breeding of the crops in China.

Description

technical field [0001] The invention belongs to the field of molecular biology and biotechnology, and relates to a wild eggplant 'TorvumVigor' (Solanum torvum Swartz) salt-tolerant gene StNHX1 gene and its herbicide-resistant plant expression vector. Background technique [0002] The threat of soil salinization to agricultural production is a global hot issue, and it is also one of the ecological crises facing the development of my country's agricultural economy. The toxic effect of salt on plants is mainly due to osmotic stress caused by water deficit and excessive Na + Toxic to important biochemical processes. The mechanism by which plants overcome salt stress is Na + efflux and vacuolar compartmentalization. Plasma membrane Na + / H + Antiporters utilize plasma membrane H + The transmembrane proton gradient generated by -ATPase converts Na + Evacuate cells, reduce Na + Long-distance transport into leaves to protect photosynthetic tissue from salt toxicity. Tonopla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/66C12N15/82C12N15/29
Inventor 朱月林盖钧镒陈国户杨立飞
Owner NANJING AGRICULTURAL UNIVERSITY