Plant heat-resistant gene HSF1 and application thereof

A plant, plant cell technology, applied in the fields of biotechnology and botany

Inactive Publication Date: 2012-01-11
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, as the genome sequencing of Chinese cabbage has not been completed and the study of functional genomics is in the ascendant, genetic breeders need to find a fast, sensitive, and efficient technique for qualitative analysis of various traits and DNA at the plant level, as well as expression at the plant level. Quantitative analysis technique of phenotype and gene expression changes for heat resistance breeding of cabbage vegetables

Method used

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  • Plant heat-resistant gene HSF1 and application thereof
  • Plant heat-resistant gene HSF1 and application thereof
  • Plant heat-resistant gene HSF1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0153] Preparation and Transformation of Agrobacterium Competent Cells by Freeze-Thawing Method

[0154] a) Pick a single colony of GV3101 from a fresh plate cultured at 28°C for 48 hours, transfer it to 20ml LB liquid medium (rif 50mg / l, GM 5050mg / l), and culture overnight at 28°C 250rpm with shaking (not too thick) . (All operations below are carried out under sterile conditions).

[0155] b) After ice bathing for 20 minutes, divide the bacteria solution into 5 ml centrifuge tubes (each tube 4 ml), and ice bath for 10 minutes.

[0156] c) Centrifuge at 4000rpm (5-10°C) for 10 minutes, and discard the bacterial solution.

[0157] d) Add 1ml of fully pre-cooled 20mM CaCl to each tube 2 To resuspend the bacteria. Ice bath for 10 minutes.

[0158] e) Centrifuge at 4000 rpm (5-10° C.) for 10 minutes, and discard the supernatant.

[0159] f) Add 300μl 20mM CaCl to each tube 2 (Depending on the concentration of the bacteria), after merging, divide into 1.5ml centrifuge tubes...

Embodiment 1

[0184] Embodiment 1, the acquisition of gene

[0185] Gene expression in plants has spatiotemporal specificity, that is, different functional gene expression occurs under different time and space conditions. The present inventors measured the expression of functional genes in Chinese cabbage samples under different heat treatment conditions by extracting mRNA from different heat-treated Chinese cabbage plants and hybridizing with a chip containing all functional genes of Chinese cabbage. Conventional methods for detecting gene expression require large-scale sequencing, which can only detect a small amount of gene expression at a time and have low detection sensitivity. Using gene chip technology can not only detect gene expression levels with high sensitivity, but also quantitatively and qualitatively detect gene expression levels, and can simultaneously study thousands of genes in the same sample. expression of thousands of genes. The use of gene chip technology can not only...

Embodiment 2

[0189] Embodiment 2, RT-PCR detects the expression of candidate heat resistance gene after heat treatment

[0190] In order to identify the function of heat-resistant genes, the inventors constructed a 35S-driven overexpression plant vector 35S::BcpHSF1 (pCAMBIA1300-HSF1), and transformed it into Arabidopsis thaliana to obtain transgenic plants OE-line 1 and OE-line2.

[0191] In order to detect the expression pattern of heat resistance genes under heat treatment conditions, the inventors used RT-PCR to measure gene expression levels. See figure 1 , it was found that the downstream target gene HSP70 of BcpHSF1 in transgenic plants 35S::BcpHSF1 was strongly expressed after heat treatment.

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Abstract

The invention relates to a plant heat-resistant gene HSF1 and application thereof. A new plant heat-resistant gene is separated from brassica plants for the first time and can improve the heat-tolerant capacity of plants obviously. The invention also provides proteins of gene codes and a preparation method thereof, a carrier and a host cell containing the gene and a method for preparing transgenic plants carrying the gene.

Description

technical field [0001] The invention belongs to the fields of biotechnology and botany; more specifically, the invention relates to a plant heat-resistant gene HSF1 and its application. Background technique [0002] Cabbage vegetables mainly include Chinese cabbage (Brassica campestris L. ssp. pekinensis) and Chinese cabbage (Brassica campestris L. ssp. chinensis). Chinese cabbage is called green cabbage for short, and it is called rapeseed in the north. It has strong adaptability, fast growth, high yield, good nutrition, and its consumption ranks first among all kinds of vegetables. There are many types and varieties of pakchoi, short growth period, wide adaptability, high yield, labor saving, easy planting, and annual production and supply. The products are tender and nutritious, and are loved by consumers. Its annual output accounts for 30-40% of the total vegetable output, making a great contribution to supplementing the off-season vegetables and achieving a balanced s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12N15/11C12P21/02C12Q1/68A01H5/00
CPCC07K14/415C12N15/8271
Inventor 何玉科孙传宝
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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