Hevea transcription factor hbmyb44 gene and its application

A transcription factor, rubber tree technology, applied in the field of genetic engineering, can solve problems such as water loss, rubber tree growth and production impact, loss, etc., to achieve the effect of small environmental impact, improved plant drought resistance, and improved tolerance.

Active Publication Date: 2018-11-06
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The rubber tree is a perennial tall tree growing in the tropics. It is the most important plant for the production of natural rubber. Its transpiration consumes a lot of water. At the same time, rubber tapping also makes it lose water. Soil moisture is the main source of water absorption. However, in recent years, my country's plantation There have been repeated droughts in Guangxi, Hainan, Yunnan and Guangdong in the rubber area, which have seriously affected and lost the growth and production of rubber trees.

Method used

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  • Hevea transcription factor hbmyb44 gene and its application
  • Hevea transcription factor hbmyb44 gene and its application
  • Hevea transcription factor hbmyb44 gene and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Cloning of embodiment 1 rubber tree transcription factor HbMYB44 gene

[0034] The rubber tree transcription factor HbMYB44 gene was cloned from the leaves of Hevea brasiliensis variety Reyan 7-33-97 by RT-PCR technology, and the specific method was as follows:

[0035] 1. Extract the total RNA in rubber tree leaves, and reverse transcribe to obtain the first strand of cDNA.

[0036] 2. Using the first strand of cDNA as a template and HbMYB44-F and HbMYB44-R as primers, the cDNA sequence of the transcription factor HbMYB44 gene was obtained by PCR amplification reaction.

[0037] The primer sequences are as follows:

[0038] HbMYB44-F: 5′-GCCAAGCTTCCATTTTATGATTACT-3′

[0039] HbMYB44-R: 5′-CTCTGTACAATGCCCAAAATTATTC-3′

[0040] Use 2×Taq PCR MasterMix (Tiangen Biochemical) for PCR amplification, the reaction system is: 2 μL of cDNA template, 1 μL of primers F and R (10 μmol / L), 25 μL of 2×PCR MasterMix, add ddH 2 0 to 50 μL.

[0041] The PCR amplification program wa...

Embodiment 2

[0044] Expression analysis of embodiment 2 HbMYB44 gene when subjected to stresses such as drought

[0045] Fluorescent quantitative PCR was used to detect the expression of HbMYB44 gene in rubber tree leaves, latex, bark and flower tissues after drought, mechanical injury, ethephon, abscisic acid and hydrogen peroxide treatments.

[0046] Drought treatment: first water the Reyan 7-33-97 budding seedlings to saturation, then put them in the light incubator for water cut-off treatment, and continuously collect 0-10d leaves as materials.

[0047] Mechanical injury treatment: the leaves of Reyan 7-33-97 were pinched with tweezers, and leaf samples were taken at 0, 0.5, 1, 2, 6 and 12 hours, and untreated plants were used as controls.

[0048] Chemical agent treatment: with brasiliensis clone CATAS7-33-97 budding seedlings as material, spraying 200 μ M abscisic acid (ABA), 1.0% ethephon (ET) and 2% hydrogen peroxide respectively at 0, 0.5, 2 , 6, 10, 24, 48 and 72 hours to take l...

Embodiment 3

[0056] Example 3 Obtaining and drought-resistant identification of transgenic Arabidopsis plants with HbMYB44

[0057] Using the HbMYB44-18T plasmid prepared in Example 1 as a template, HbMYB44-OE-F: 5'-CCC AAGCTT ATGGCTGTTACT-3' (the underline is the HindIII restriction site) and HbMYB44-OE-R:5'-CG GGATCC CTCAATCTTACT-3' (the underline is the BamHI restriction site) was used as a primer to amplify the ORF of the HbMYB44 gene, and the amplified product was digested by HindIII and BamHI and connected to the expression vector after the same digestion (the transformed pGreenⅡ vector, That is, the mGFP-6×His fragment was inserted between the SpeI and XbaI sites of the original vector backbone), the constructed recombinant plasmid was named HbMYB44-OE-GFP, and the recombinant plasmid HbMYB44-OE-GFP was transformed into Agrobacterium GV3101 strain by electric shock method ; The HbMYB44 gene was transformed into wild-type Arabidopsis plants (Col) by the Agrobacterium-mediated metho...

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Abstract

The invention relates to a rubber tree transcription factor HbMYB44 gene and an application thereof. The transcription factor gene HbMYB44 related to the drought resistance of the rubber tree provided by the present invention, through transforming the Arabidopsis thaliana with the HbMYB44 gene to carry out functional research on the gene, verifies that the drought stress tolerance of the transgenic Arabidopsis is improved. The gene can be used in drought-resistant transgenic research of rubber trees and other crops. The transcription factor gene HbMYB44 related to the drought resistance of the rubber tree in the invention is from the rubber tree, does not generate gene drift, has little impact on the environment, and provides new gene resources for improving the drought resistance of plants.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the rubber tree transcription factor HbMYB44 gene and its application. Background technique [0002] The rubber tree is a perennial tall tree growing in the tropics. It is the most important plant for the production of natural rubber. Its transpiration consumes a lot of water. At the same time, rubber tapping also makes it lose water. Soil moisture is the main source of water absorption. However, in recent years, my country's plantation There have been repeated droughts in Guangxi, Hainan, Yunnan and Guangdong in the rubber area, which have seriously affected and lost the growth and production of rubber trees. Therefore, in-depth research on the drought resistance of rubber trees and the development and breeding of drought-resistant strains are of great significance to the rational allocation of strains in my country's rubber planting areas, as well as to the utilizati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/20C12Q1/6895C12N15/11
CPCC07K14/415C12N15/8273C12Q1/6895
Inventor 王立丰王纪坤安锋谢贵水
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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