Application of rice genes OsVHA1 in delaying senility of plant leaves and improving plant salt tolerance

A plant leaf and gene technology, applied in angiosperms/flowering plants, botanical equipment and methods, applications, etc., can solve the problems of female gametophyte death, more leaf division, and so on, so as to delay the aging of plant leaves, The effect of improving salt tolerance

Inactive Publication Date: 2015-07-01
ZHEJIANG UNIV
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Problems solved by technology

However, existing studies have shown that it is only involved in the regulation of some traits. For example, the loss of function of subunit A in Arabidopsis can lead to complete male sterility and lethality of some female gametophytes (Dettmer J, Schubert D, Calvo-Weimar O, Stierhof YD, Schmidt R, Schumacher K. Essential role of the V-ATPase in male gametophyte development. Plant J. 2005, 41: 117-122), while antisense inhibition of carrot A subunit resulted in smaller cells and more leaf division Multiple (Gogarten J P, Fichmann J, Braun Y, et al. The use of antisense mRNA to inhibit the tonoplast H+-ATPase in carrot. The Plant Cell, 1992, 4: 851-864), antisense inhibits the A1 subunit of rice then only lead to increased leaf stomatal density and sensitivity to salt and drought (Zhang H, Niu X, Liu J, Xiao F, Cao S, Liu Y. RNAi-directed downregulation of vacuolar H(+)-ATPase subunit a results in enhanced stomatal aperture and density in rice.PLOS ONE.2013,8:e69046), and overexpression of apple A subunit in tobacco can improve the salt tolerance of transgenes (Dong QL, Wang CR, Liu DD, Hu DG, Fang MJ , You CX, Yao YX, Hao YJ. MdVHA-A encodes an apple subunit A of vacuolar H(+)-ATPase and enhances drought tolerance in transgenic tobacco seedlings. J Plant Physiol. 2013, 170: 601-609)
[0006] So far, although many existing studies have shown that the A subunit plays a very important role in plant growth and development, there has been no report on its regulation of plant leaf senescence

Method used

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  • Application of rice genes OsVHA1 in delaying senility of plant leaves and improving plant salt tolerance
  • Application of rice genes OsVHA1 in delaying senility of plant leaves and improving plant salt tolerance
  • Application of rice genes OsVHA1 in delaying senility of plant leaves and improving plant salt tolerance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Isolation and genetic analysis of mutants

[0029] pass 60Co radiation mutagenesis long-lasting high blast resistance material self-selection 1 (from the Rice Research Institute of Enshi Academy of Agricultural Sciences), screened out the premature leaf senescence and salt-sensitive mutant osvha1 from the offspring, and obtained stable mutant traits through multiple generations of backcrossing and selfing strains. The premature senescence trait starts at the tillering stage of osvha1 mutant plants ( figure 1 A and 1B), by the time of heading and flowering stage, all leaves showed different degrees of premature senescence symptoms ( figure 1 C). Using osvha1 as the female parent, the F 1 , F 1 The plants behaved normally; F 1 F obtained by selfing 2 The offspring showed wild type and mutant phenotypes, and the segregation ratio of wild type:mutant=3:1, thus indicating that the senescence traits were controlled by a single recessive gene.

Embodiment 2

[0030] Example 2 Salt Sensitivity Analysis of Mutant osvha1

[0031] at 100mmol L -1 After being treated for 7 days under NaCl stress, the leaves of the osvha1 mutant were curled, and the tips of the leaves were partially dead ( figure 2 A). Simultaneously, plant height and biological yield are compared with embodiment 1 described wild-type control self-selected No. 1 and decline respectively 10.6% and 29.0% ( figure 2 A), while the 7d plant net growth height, root net growth length and net biological yield were respectively 13.7%, 9.9% and 37.5% lower than the control. After returning to growth for 5 days, 12.4% of the mutants withered and died, and the growth rate of the treated mutants was significantly slower than that of the treated wild-type control ( figure 2 B), the mutant plant height and biological yield are respectively 43.5% and 53.8% less than the control, while the net growth height of the 5d plant, the net growth length of the root and the net biological y...

Embodiment 3

[0032] Example 3 Fine mapping of aging gene OsVHA1

[0033] 500 pairs of SSR markers and 50 pairs of InDel markers evenly distributed on 12 chromosomes of rice were selected, and polymorphism screening was performed on the osvha1 mutant and 02428 one by one. The results showed that 178 pairs of primers were polymorphic between the two parents. Sex ratio was 32.4%. Then these 178 pairs of markers were used to conduct BSA analysis on the two constructed gene pools, and it was found that three SSR markers RM20361, RM20491 and RM3430 on the long arm of rice chromosome 6, and one InDel marker R6M44 were in the mutant pool and There is a clear partial separation between normal pools. Subsequently, the 50 F 2 Genotype analysis was performed on individual plants with mutant phenotypes, and the physical arrangement position of the four markers and the OsVHA1 gene on chromosome 6 was determined in combination with the genotype of each molecular marker as RM20361-R6M44-RM20491-OsVHA1-R...

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Abstract

The invention discloses application of rice genes OsVHA1 in delaying senility of plant leaves and improving plant salt tolerance. A nucleotide sequence of the rice genes OsVHA1 is shown in SEQ ID NO.1; the application comprises the processes that the rice genes OsVHA1 are joined with an expression carrier to construct and obtain a recombinant expression carrier; the recombinant expression carrier is converted into a receiver plant. Base on a tula cloning technology, the cloning from a rice premature senescence mutant osvha1 is carried out to obtain genes OsVHA1; a functional complementary experiment proves that the genes OsVHA1 can delay the senility of plant leaves while a salt processing experiment proves that the genes OsVHA1 can improve the salt tolerance capability of the plant; the genes OsVHA1 can be applied to the field of plant breeding and are significant in screening and selective breeding of plant varieties with senescence resistance and salt tolerance.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering and rice molecular breeding, in particular to the application of a rice gene OsVHA1 in delaying plant leaf senescence and improving plant salt tolerance. Background technique [0002] Rice is one of the most important food crops, and more than half of the world's population depends on rice as a staple food. In our country, with the continuous increase of population and the decrease of land area, the demand for increasing rice production is becoming more and more urgent, so the national food security strategy is particularly important. Therefore, cultivating high-yield and high-quality rice varieties and continuously increasing rice yields are the most important tasks to ensure my country's food security. Leaf is the most important photosynthetic organ of rice. According to theoretical calculations, 60% to 80% of the nutrients in rice grain filling come from the photosynthesis of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C12N1/21A01H5/00
Inventor 潘刚龚盼黎坤瑜杨茜程方民
Owner ZHEJIANG UNIV
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