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Application of rubber tree glutaredoxin HbGRX to field of selection of rubber tree tapping panel dryness resistant varieties

A rubber tree and protein technology, applied in the field of plant genetic engineering, can solve the problems of uneven control effect, unstable control effect, increase the workload of rubber trees, etc., and achieve the effect of removing a large amount of accumulation

Inactive Publication Date: 2019-03-15
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

[0006] In the prior art, rubber tree dead skin is often prevented and treated by adding rehabilitation nutrients. On the one hand, it increases the workload of rubber tree planting, and the effect of prevention and treatment is also uneven, and the effect of prevention and control is not stable. There are few reports on the prevention and control technology of rubber tree dead skin
In addition, if the selection of rubber tree species with anti-dead skin performance can be carried out from the molecular level, and the rubber tree species with better anti-dead skin performance can be selected and cultivated, it will have positive significance for the development of the rubber industry. There are few reports on the molecular breeding technology of dead bark rubber tree varieties

Method used

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  • Application of rubber tree glutaredoxin HbGRX to field of selection of rubber tree tapping panel dryness resistant varieties
  • Application of rubber tree glutaredoxin HbGRX to field of selection of rubber tree tapping panel dryness resistant varieties
  • Application of rubber tree glutaredoxin HbGRX to field of selection of rubber tree tapping panel dryness resistant varieties

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Total RNA Extraction and cDNA First Strand Synthesis

[0045] The method (Tang C., Qi J., Li H., Zhang C., Wang Y., 2007.A convenient and efficient protocol for isolating high- quality RNA from latex of Heveabrasiliensis (para rubbertree). J. Biochem. Biophys. Methods 70(5), 749-754.).

[0046] For the extraction of total RNA from materials other than latex, the plant total RNA extraction kit from TIANGEN Company was used, and the operation method was referred to the kit instruction manual.

[0047] The first strand of cDNA was synthesized using the Fementas Reverse Transcription Kit (RevertAid First Strand cDNA Synthesis Kit) according to the company's operating instructions.

Embodiment 2

[0048] Bioinformatics analysis of embodiment 2HbGRX gene

[0049] In previous studies, a hevea glutaredoxin HbGRX (accession number ABZ88803.1 / EU295478.1) whose expression was significantly down-regulated in dead bark trees (compared with healthy trees) has been identified from the proteomic level. The amino acid sequence is shown in SEQ ID No.2, and the gene sequence encoding the Hevea glutaredoxin HbGRX is shown in SEQ ID No.1.

[0050] Use the ExPasy online tool ProParam (http: / / www.expasy.org / tools) to analyze the various amino acid content, theoretical molecular weight and isoelectric point of HbGRX protein and other physical and chemical parameters;

[0051] Use NCBI's CDD (Conserved Domain Database) database (http: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi) to predict the conserved domain of the protein;

[0052] Use SignalP 4.1Server (http: / / www.cbs.dtu.dk / services / SignalP / ) to predict the signal peptide of the protein;

[0053] Use TMHMM Server v.2.0 (http: / / www...

Embodiment 3

[0065] The subcellular localization of embodiment 3HbGRX protein

[0066] Primers 1302-HbGRX-F and 1302-HbGRX-R (shown in Table 1) for subcellular localization were designed according to the sequence of the target gene HbGRX, and normal latex cDNA was used as a template for PCR amplification, and the PCR product and the pCAMBIA1302 vector were respectively Carry out double digestion with restriction endonuclease NcoI / SpeI, recover the digested product, and then use T4-DNA ligase to connect the target fragment and the vector fragment to obtain the plant expression vector pCAMBIA1302-HbGRX-GFP, and transform the large intestine according to the conventional method After the Bacillus BL21 competent cells were selected, single clones were picked for expansion and cultured. After verification by PCR and plasmid double enzyme digestion, sequencing analysis was performed, and the sequenced correct positive clone plasmid (pCAMBIA1302-HbGRX-GFP) and empty vector (pCAMBIA1302-GFP) Trans...

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Abstract

The invention belongs to the technical field of plant gene engineering, and particularly relates to rubber tree glutaredoxin HbGRX, and further discloses application of the rubber tree glutaredoxin HbGRX to the field of selection of rubber tree tapping panel dryness resistant varieties. According to the method, the fact that the expression of the rubber tree glutaredoxin HbGRX in a rubber tree with tapping panel dryness shows up an obvious reducing trend is verified through gene cloning performed on the rubber tree glutaredoxin HbGRX and analysis and research on biological information and expression performance, visibly GRX as antioxidant enzyme can effectively eliminate a large amount of accumulated laticiferous cell active oxygen, has close correlation with rubber tree tapping panel dryness, can be effectively applied to the field of prevention and control of rubber tree tapping panel dryness, lays a powerful theoretical foundation for illuminating a rubber tree tapping panel drynessoccurrence mechanism, and further provides a scientific basis for the research and development of a rubber tree tapping panel dryness prevention and control technology and breeding of tapping panel dryness resistant rubber tree new variety through molecular selection.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, specifically relates to a rubber tree glutaredoxin HbGRX, and further discloses its application in the fields of rubber tree dead skin resistance breeding and rubber tree dead skin prevention and control. Background technique [0002] During the growth and development of plants, they will be subjected to various biotic and abiotic stresses, such as pathogen attack, photooxidation, heavy metals, drought, extreme temperature, etc. These stresses will stimulate the production of reactive oxygen species (ROS) in plants to a certain extent. ), and excessive accumulation of reactive oxygen species will cause oxidative damage to proteins, DNA and lipids, and then lead to cell death (Apel and Hirt, 2004). In order to cope with this kind of oxidative stress, plants will evolve a complex antioxidant system to eliminate the damage caused by ROS and realize their own protection and repair. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N15/53C12N15/82A01H5/04A01H6/38C12Q1/6895
CPCC12N9/0004C12N15/8261C12Q1/6895C12Q2600/13C12Q2600/158C12Y120/04001
Inventor 袁坤罗萍张华林郭秀丽冯成天胡义钰王真辉
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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