Application of Phospholipase pldζ1 Gene in Improving Salt Tolerance of Plants

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: 2019-12-06
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
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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 pldζ1 Gene in Improving Salt Tolerance of Plants
  • Application of Phospholipase pldζ1 Gene in Improving Salt Tolerance of Plants
  • Application of Phospholipase pldζ1 Gene in Improving Salt Tolerance of Plants

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Experimental program
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Effect test

Embodiment 1

[0030] PLDζ1 Gene Cloning and Overexpression Enhance 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, and then the cDNA was used as a template, using PLDζ1 sequence-specific primers PLDζ1FZ: 5'-GGGGTACCATGCAAGAATATCTGAACC-3' and PLDζ1RZ: 5'-CGGGATCCATGGAAAACTTGTGGAG-3' was amplified by PCR, and the amplified target fragment included the sequence shown in SEQ ID NO.1. The amplified target fragment was inserted into the restriction site of the plant overexpression vector pU1301D1 (modified by pCAMBIA1301, Hajdukiewicz et al., The small, versatile pPZPfamily of Agrobacterium binary vectors for plant transformation. PlantMol. Biol. 1994, 25: 989-994) Between Kpn I and BamH I, the pU1301D1:PLDζ1 recombinant plasmid was obtained. After ...

Embodiment 2

[0044] Overexpression of PLDζ1 enhances salt tolerance of plants

[0045] Two PLDζ1-OE independent strains OE-14 and OE-15 of the PLDζ1 overexpression rice transformed plants created in Example 1 were randomly selected for detailed analysis, and the non-transformed regenerated plants (wild type, WT) that had also undergone the tissue culture process were randomly selected. As a control, it was treated with different concentrations of NaCl solution (0, 50, 75mM) for one month at the tillering stage. The results found that under normal conditions, PLDζ1 overexpressed plants had no significant difference in plant height, leaf plasma membrane permeability, and malondialdehyde content between the corresponding wild-type plants. significantly higher than that of the wild type, and its leaf plasma membrane permeability and malondialdehyde content were significantly lower than those of the non-transformed wild type ( figure 1 ). After one month of 75mM NaCl treatment, the malondiald...

Embodiment 3

[0047] Isolation, Identification and Functional Identification of PLDζ1 Deletion Mutant

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

[0049] In order to further verify and analyze the function of the 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 into It 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 there was a small amount of expression of PLDζ1 in the pldζ1-1 homozygous mutant, but it was significantly lower than that of the wild type, while in the pldζ1-2 homozygous mutant PLDζ1mRNA could not be detected, which belonged to complete deletion mutant.

[0050] The analysis show...

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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, and specifically relates to the application of phospholipase PLDζ1 gene in improving plant salt tolerance and increasing crop yield under salt stress. Background technique [0002] With the change of climate and environment and the continuous increase of population, the problem of global food security is worrying. In recent years, the frequent occurrence of drought and the continuous expansion of drought and salinized cultivated land have made the environmental stress of crop production more and more serious. Land salinization is one of the main problems faced by agricultural production. 1 / 5 of the cultivated land in the world is salinized (Yamaguc hi et al., Developing salt-tolerant crop plants: challenges and opportunities. Trends Plant Sci. 2005, 10:615-620.), and 1 / 3 of the farmland in my country is too salty As a result, crop yields have decreased, and the occurrenc...

Claims

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Application Information

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Patent Type & Authority Patents(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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