Torch pear haloduric gene PpGST and application thereof

A salt-tolerant gene, torch pear technology, applied in the fields of molecular biology and genetic engineering, can solve the problems affecting crop yield and quality, agricultural production loss, long cycle, etc., achieve broad market application prospects, reduce yield reduction, and short breeding cycle Effect

Inactive Publication Date: 2010-06-23
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The salinization of soil seriously affects the yield and quality of crops, causing significant losses to agricultural production
At present...

Method used

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  • Torch pear haloduric gene PpGST and application thereof
  • Torch pear haloduric gene PpGST and application thereof
  • Torch pear haloduric gene PpGST and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0020] 1) Cloning and sequence analysis of the full-length PpGST gene

[0021] Grind the red peel of the torched pear into powder with liquid nitrogen, transfer it to a centrifuge tube, and extract total RNA by the guanidine isothiocyanate method. Use reverse transcriptase M-MLV (Tiangen) to synthesize the first strand of cDNA using total RNA as a template. The reaction system and operation process are as follows: Take 5 μg Total RNA, add 50ng oligo(dT)15, 2μL dNTP (2.5mM each) in sequence ,ddH 2 O (RNase-free) to a reaction volume of 13.5 μL; after mixing, heat denaturation at 70°C for 5 minutes, then rapidly cool on ice for 5 minutes, then add 4 μL 5×First-stand buffer, 0.5 μL RNasin (200 U), 1 μL M - MLV (200U), 1 μL DTT (0.1M), mix well and centrifuge briefly, incubate at 42°C for 1.5h, take it out and heat at 95°C for 5min to terminate the reaction. After the first strand of cDNA was synthesized, it was stored at -20°C for future use.

[0022] Using the synthesized fir...

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Abstract

The invention relates to a torch pear haloduric gene PpGST and an application thereof. The PpGST gene is provided with base sequence as SEQ ID indicates and encoded glutathione S-transferring enzyme. In the invention, the PpGST gene is proven to have the function of improving plant resisting salt stress by the technology related to functional genomics; and when the haloduric gene PpGST is structured on a plant expression vector and shifted to over expression in tobacco, the obtained transgenic tobacco enjoys very strong haloduric activity.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic engineering, in particular to a new salt-tolerant gene PpGST obtained from Yunnan torched pear (Pyrus pyrifolia Nakai) and its application. Background technique [0002] With the rapid development of industrial and agricultural production and the continuous increase of population, the society's demand for food is increasing day by day, but the shortage of cultivated land resources is becoming more and more serious. Soil erosion, salinization, desertification, degradation and pollution have always been the five major soil problems plaguing human beings. According to data from the Food and Agriculture Organization of the United Nations, the total area of ​​salinized land in the world is about 9.5×10 8 hectares, accounting for 7.26% of the earth's land area. The area of ​​salinized land in my country is also very large, about 1.0×10 8 hectares, of which modern salinized land accounts ...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/84
Inventor 刘迪秋王继磊葛锋方松刚李文娴田荣欢王光勇
Owner KUNMING UNIV OF SCI & TECH
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