Application of phospholipase gene PLDzeta1 in improving salt resistance of plant

A technology of salt tolerance and genetics, applied in the field of plant molecular breeding and biology, can solve problems such as complex molecular mechanisms

Active Publication Date: 2016-12-07
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, salt stress and drought are quantitative traits controlled by multiple genes (Denby et al., Eng ineering drought and salinity tolerance implants: lessons from genome-wide expression pro filing in Arabidopsis. Trends Biotechnol. 2005, 23:547-552; Sahi et al., Salt stress response in rice: genetics, molecular biology, and comparative genomics. Funct Integr Genomics. 2006, 6: 263-284), is regulated by multiple signaling or metabolic pathways, and each pathway has an interactive relationship, and its molecular mechanism is equivalent Complex, more challenging breeding for drought and salt tolerance

Method used

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  • Application of phospholipase gene PLDzeta1 in improving salt resistance of plant
  • Application of phospholipase gene PLDzeta1 in improving salt resistance of plant
  • Application of phospholipase gene PLDzeta1 in improving salt resistance of plant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Cloning of PLDζ1 Gene and Its Overexpression Enhances Plant Salt Tolerance

[0031] 1. Cloning of full-length cDNA of PLDζ1 gene and construction of plant overexpression vector:

[0032] Total RNA was extracted from the leaves of japonica rice Dongjin, and the first-strand cDNA was synthesized by reverse transcription RT-PCR using mRNA as a template. Then, using the cDNA as a template, the sequence-specific primers PLDζ1FZ: 5'-GGGGTACCATGCAAGAATATCTGAACC-3' and PLDζ1RZ were used: 5'-CGGGATCCATGGAAAACTTTGGGAG-3' was amplified by PCR, and the target fragment obtained by amplification included the sequence shown in SEQ ID NO.1. The amplified target fragment was inserted into the enzyme cleavage site of the plant overexpression vector pU1301D1 (modified by pCAMBIA1301, Hajdukiewicz et al., The small, versatile pPZPfamily of Agrobacterium binary vectors for plant transformation. Plant Mol. Biol. 1994, 25: 989-994) Between Kpn I and BamH I, the pU1301D1:PLDζ1 recombinant pla...

Embodiment 2

[0044] PLDζ1 overexpression enhances salt tolerance in plants

[0045] Two PLDζ1-OE independent lines, OE-14 and OE-15, of the PLDζ1-overexpressing rice transformed plants created in Example 1 were randomly selected for detailed analysis, and non-transformed regenerated plants (wild type, WT) that had also undergone the tissue culture process were selected. As a control, they were treated with different concentrations of NaCl solutions (0, 50, 75 mM) for one month during the tillering stage. The results showed that under normal conditions, there was no significant difference in plant height, leaf plasma membrane permeability and malondialdehyde content between PLDζ1 overexpressing plants and the corresponding wild type, but under salt stress conditions, the survival rate of PLDζ1 overexpressing plants significantly higher than the wild type, and its leaf plasma membrane permeability and malondialdehyde content were significantly lower than those of the non-transformed wild typ...

Embodiment 3

[0047] Isolation, Characterization and Gene Function Characterization of PLDζ1 Deletion Mutants

[0048] 1. Isolation, identification and phenotype observation of mutant pldζ1:

[0049] In order to further verify and analyze the function of PLDζ1 gene, the present invention isolated and obtained two independent mutant lines with T-DNA inserted into different sites of PLDζ1, named pldζ1-1 and pldζ1-2 respectively, wherein the T-DNA of pldζ1-1 was inserted is located in the 3'-UTR region of PLDζ1, while the T-DNA insertion of pldζ1-2 is located in the third intron of PLDζ1. The effect of T-DNA insertion on the expression of PLDζ1 was analyzed by semi-quantitative RT-PCR. The results showed that the expression of PLDζ1 in the homozygous mutant of pldζ1-1 was slightly lower than that of the wild type, while that in the homozygous mutant of pldζ1-2 PLDζ1mRNA could not be detected, which belonged to the complete deletion mutant.

[0050] The analysis showed that PLDζ1 was induced ...

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Abstract

The invention discloses application of a phospholipase gene PLDzeta1 in improving salt resistance of a plant. The gene PLDzeta1 is transferred to a plant through a molecular genetic transformation method, and the transformation plant with the overexpression gene PLDzeta1 is obtained. The overexpression gene PLDzeta1 remarkably improves the salt resistance of the plant. The membrane permeability and the malondialdehyde content of a leaf of the plant with overexpression gene PLDzeta1 are remarkably lower than those of a control wide group under the threat of salt, and the survival rate and the plant height are remarkably higher than those of the control wide group. The loss expression of the gene PLDzeta1 leads to remarkable reduction of the salt resistance of the plant and remarkable reduction of biological yield compared with the wild group, and the result further improves that the gene PLDzeta1 has a remarkable positive regulation effect on salt resistance. The application of the phospholipase gene PLDzeta1 in improving salt resistance of the plant provides a novel way for the seed breeding of salt-resistance molecules of the plant, and creates a novel seed material.

Description

technical field [0001] The invention belongs to the field of plant molecular breeding and biotechnology, in particular to the application of phospholipase PLDζ1 gene in improving plant salt tolerance and increasing crop yield under salt stress. Background technique [0002] With the changing climate and environment, and the increasing population, global food security is a concern. In recent years, the frequent occurrence of drought and the continuous expansion of the area of ​​drought and salinized cultivated land have made the problem of environmental stress in crop production more and more serious. Land salinization is one of the main problems faced by agricultural production. One-fifth of the world's arable land is salinized (Yamaguc hi et al., Developing salt-tolerant crop plants: challenges and opportunities. Trends Plant Sci. 2005, 10: 615-620.), and one-third of the farmland in my country is too salty This has led to the reduction of crop yields, and the occurrence o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N15/55A01H5/00
CPCC12N9/16C12N15/8205C12N15/8273C12Y301/04
Inventor 洪月云吕玮鑫孙林啸
Owner HUAZHONG AGRI UNIV
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