Application of purple acid phosphatase gene GmPAP35 of soybean

A technology of acid phosphatase and soybean, which is applied in the field of genetic engineering to achieve the effect of improving soybean phosphorus efficiency, promoting carbon metabolism ability, and improving plant phosphorus efficiency

Active Publication Date: 2018-09-28
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the difference is that the expression of AtPAP2 is not induced by low phosphorus, while the expression of GmPAP35 is induced by low

Method used

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  • Application of purple acid phosphatase gene GmPAP35 of soybean
  • Application of purple acid phosphatase gene GmPAP35 of soybean
  • Application of purple acid phosphatase gene GmPAP35 of soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 GmPAP35 gene expression pattern analysis

[0025] By quantitative PCR technology, it was confirmed that GmPAP35 gene was strongly expressed by low phosphorus. The length of the open reading frame of the GmPAP35 gene is 1989bp, encoding a protein of 662 amino acids.

[0026] The plant materials are planted in greenhouse nutrient solution hydroponic cultivation. First, seeds with full grains and complete seed coats are selected, and after the seeds are sterilized by chlorine gas fumigation (100mL sodium hypochlorite + 4mL concentrated HCl) for 13.5hrs, the seeds are germinated by the rolling paper tube method. After the cotyledons are fully unfolded, the healthy and consistent seedlings are moved to the soybean nutrient solution with a pH of about 5.8. Two weeks later, when the seedlings grow the first complete three compound leaves, the phosphorus level treatment is carried out. Phosphorus treatments included low phosphorus (5 μM) (LP) and high phosphorus (...

Embodiment 2

[0036] Cloning of embodiment 2GmPAP35 gene

[0037] Using the soybean leaf cDNA as a template, the upstream specific primer 5′- CTCGAG ATGATTCCCGATCTACCCCTC-3'(SEQ ID NO:5) and downstream specific primer 5'- ACTAGT TCAAGTTTCCTCAGTCTTCAC-3' (SEQ ID NO: 6) amplified the full-length ORF sequence of 1989bp of the GmPAP35 gene, and sequenced and compared the obtained GmPAP35 coding sequence, see Glyma20g03260.1, and the corresponding protein sequence, see Glyma20g03260.1.

Embodiment 3

[0038] Example 3 Overexpression GmPAP35 vector construction

[0039] Construction of overexpression vector: using soybean leaf cDNA as template, using upstream specific primer 5'- CTCGAG ATGATTCCCGATCTACCCCTC-3'(SEQ ID NO:5) and downstream specific primer 5'- ACTAGT TCAAGTTTCCTCAGTCTTCAC-3′(SEQ ID NO:6) amplifies the full-length ORF sequence of GmPAP35 gene 1989bp, after the PCR fragment is recovered and sequenced correctly, the fragment and the target vector are digested with Xho I and Spe I, and the GmPAP35 gene is connected to the destination vector pTF101.1.

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Abstract

The invention discloses the application of a purple acid phosphatase gene GmPAP35 of a soybean, and belongs to the field of gene engineering. Through functional analysis of the GmPAP35 in a whole conversion material of the soybean, a function of the GmPAP35 participating in carbon metabolism in a plant is specified. Overexpression of the GmPAP35 can increase the height of a soybean plant and the contents of glucose and fructose in the plant body, so that the soybean biomass and the phosphorus uptake are increased, and the soybean phosphorus efficiency is improved; a theoretical basis is laid for culturing a new variety of phosphorus efficient-transgenic crops. The purple acid phosphatase gene GmPAP35 has the capacity of promoting the carbon metabolism of the crops through a transgenic technology, so that the plant phosphorus efficiency is improved, and new purposes of fertilizer use amount reduction and high yield are achieved. The purple acid phosphatase gene GmPAP35 has an importanttheoretical and practical significance on developing environmentally-friendly sustainable agriculture.

Description

technical field [0001] The present invention relates to new functions and new applications of known genes. Specifically, the invention relates to the application of the purple acid phosphatase gene GmPAP35 in promoting soybean carbon metabolism, cultivating phosphorus-efficient transgenic legumes, and improving crop phosphorus efficiency, which belongs to the field of genetic engineering. Background technique [0002] Phosphorus is one of the essential nutrients in the process of plant growth and plays a very important role in the whole metabolic process of plants. However, phosphorus is easily fixed by some metal ions in the soil to form insoluble compounds that are difficult to be directly absorbed and utilized by plants, often resulting in a lack of available phosphorus in the soil, which severely limits the growth of crops (Raghothama, 1999; Vance et al., 2003). During the long-term evolutionary process, plants have developed many adaptive mechanisms to improve phosphor...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/54
CPCC12N15/8245C12N15/8261
Inventor 王秀荣张海燕李成晨杨韬
Owner SOUTH CHINA AGRI UNIV
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