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A method for increasing plant tolerance to NaCl by downregulating Pab2 and Pab4

A plant and salt tolerance technology, applied in the fields of botanical equipment and methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as limited effect of salt tolerance, and achieve the effect of improving genetic characteristics

Inactive Publication Date: 2019-02-05
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Salt tolerance in plants is a complex trait controlled by polygenic quantitative traits and influenced by the environment; therefore, traditional breeding methods have limited effect on improving crop salt tolerance

Method used

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  • A method for increasing plant tolerance to NaCl by downregulating Pab2 and Pab4
  • A method for increasing plant tolerance to NaCl by downregulating Pab2 and Pab4
  • A method for increasing plant tolerance to NaCl by downregulating Pab2 and Pab4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Identification and expression analysis of pab2-1 mutant, pab4-1 mutant and pab2-1pab4-1 double mutant

[0058] 1. Identification of pab2-1 mutant, pab4-1 mutant and pab2-1pab4-1 double mutant

[0059] according to figure 1 The primer combination in the pab2-1pab4-1 double mutant homozygous plants were identified by PCR technology; in addition, the pab2-1 mutant and pab4-1 mutant homozygous plants were identified by the same method.

[0060] The primer sequences used are as follows:

[0061] pab2-1LP: 5'-ATTCGAAAGTGTCAAACACGC-3'(Left primer, LP)

[0062] pab2-1RP: 5'-TAATAAAAATGTTGCCAGCGC-3'(Right primer, RP)

[0063] pab4-1LP: 5'-GAGAAGGCAATGCAGAAGTTG-3'(Left primer, LP)

[0064] pab4-1RP: 5'-ACTACCTGCAGCTTAGCCCTC-3'(Right primer, RP)

[0065] LBb1: 5'-GCGTGGACCGCTTGCTGCAACT-3'(Left border primer, LB)

[0066] 2. Analysis of corresponding gene expression in pab2-1 mutant, pab4-1 mutant and pab2-1pab4-1 double mutation

[0067] Total RNA was extracted f...

Embodiment 2

[0088] Example 2, analysis test of pab2-1pab4-1 double mutant tolerance to NaCl

[0089] NaCl can inhibit the growth of plant seedlings. With the increase of exogenous NaCl concentration, the inhibition of Arabidopsis wild type (Col-0 ecotype) was enhanced, and the growth of its main root and leaves were affected to varying degrees. The experiment was repeated three times.

[0090] Arabidopsis wild type (Col-0 ecotype) and pab2-1pab4-1 double mutant plants were used as experimental materials. Seeds of each experimental material were sown on MS plates without NaCl, stratified at 4°C for 3 days, then placed in a light incubator for 72 hours, and then the newly germinated seedlings of each experimental material were transferred to MS plates containing different concentrations of NaCl. On the medium (0mM, 50mM, 100mM, 150mM and 200mM), observe the growth of Arabidopsis seedlings after vertical growth for 9 days and take pictures.

[0091] The result is as Figure 5 As shown, on t...

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Abstract

The invention discloses a method for improving tolerance of plants for NaCl by reducing the content of PAB2 and PAB4. The method specifically provides an application of protein 1 and protein 2 serving as targets to any one of the following conditionsthat a1, salt tolerance of the plants is improved; a2, plant varieties with improved salt resistance are bred. Protein 1 is PAB2, and protein 2 is PAB4. It is found through the research that tolerance of PAB2 and PAB4 gene double knockout mutants obtained in the arabidopsis thaliana bioresearch center to NaCl is enhanced; thus, the tolerance of the plants to NaCl can be adjusted through PAB2 and PAB4; in addition, PAB2 and PAB4 low-expression and salt-resistant transgenic crops can be obtained through the gene engineering means (PNAi technology). The method meets the sustainable agriculture development demand, and high practical value and market prospects are achieved on the aspects of researching the molecular mechanism for plants to tolerant NaCl, improving hereditary characters, cultivating efficient salt-resistant new varieties and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method for improving plant tolerance to NaCl by down-regulating PAB2 and PAB4. Background technique [0002] As the world's population continues to grow, the human demand for food is increasing. Salt stress is one of the main factors affecting crop yields worldwide. Salt stress mainly causes damage to plants through sodium chloride as a stress. The high concentration of sodium ions in the soil will directly affect the protein synthesis, photosynthesis and energy metabolism of plants, etc., resulting in stunted plant growth, reduced germination rate, inhibited growth and differentiation of organs, slowed metabolism, reduced protein synthesis, etc., ultimately affecting crop yield. So far, the total area of ​​saline-alkali land in the world has reached 840 million hm 2 , accounting for about 6% of the world's land area, and 100-400 million hm per year 2 The speed increases; the are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 张大鹏卢艳芬于泳涛王晓静王小芳
Owner TSINGHUA UNIV
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