SEC12-like protein gene CPU1 and application thereof in improving soybean phosphorus efficiency

A sec12-like, CPU1-H2 technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of not being cloned into soybean phosphorus efficiency main gene

Pending Publication Date: 2022-01-14
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are few reports on the analysis of the genetic basis of the natural variation of soybean phosphorus efficie

Method used

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  • SEC12-like protein gene CPU1 and application thereof in improving soybean phosphorus efficiency
  • SEC12-like protein gene CPU1 and application thereof in improving soybean phosphorus efficiency
  • SEC12-like protein gene CPU1 and application thereof in improving soybean phosphorus efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1, Genetic Mapping and Candidate Gene Identification of Phosphorus Uptake Efficiency

[0028] The present invention utilizes a set of core populations of soybean phosphorus efficiency germplasm resources (including 274 soybean materials) collected in the early stage to carry out field experiments in the experimental field of Boluo, Guangdong (East longitude 113°50′, North latitude 23°07′). Block design (1.5 m per plot 2 ), 4 blocks were set up, and the phenotype identification was carried out for the indicators related to phosphorus efficiency.

[0029] Determination of phosphorus content: phosphorus content (mg / plant) = phosphorus concentration (mg / g) × plant dry weight (g / plant), where phosphorus concentration is measured by colorimetry (Murphy and Riley, 1963).

[0030]Determination of total root length: In order to obtain a complete plant root system, use tools such as shovel to dig down in a square area of ​​40 cm × 40 cm (with the plant as the center), re...

Embodiment 2

[0037] Embodiment 2, CPU1 Cloning and functional verification of

[0038] according to CPU1 The cDNA sequence of the gene (as shown in SEQ ID NO.2) designed specific primers F1 / R1, using the cDNA sample of the wild-type soybean variety YC04-5 as a template to amplify a 147 bp fragment; using Swa Ⅰ+ Asc Ⅰ Digest the above 147bp fragment to obtain the forward fragment, and then clone the forward fragment into the pFGC5941 vector Swa I and Asc between Ⅰ; use SM a I+ Bam H I digested the above 147 bp fragment to obtain the reverse fragment, and then cloned the reverse fragment into the pFGC5941 vector that already contained the forward fragment SM a I and Bam The recombinant vector was obtained between HI, and the recombinant vector was transformed into Agrobacterium EHA105 competent cells, and the bacteria were shaken for subsequent use. Agrobacterium tumefaciens-mediated cotyledon node transformation method (Wang et al . 2009) to obtain CPU1 whole plant interv...

Embodiment 3

[0049] Embodiment 3, CPU1 Variations in amino acid sequence and expression

[0050] identified by genome-wide association analysis CPU1 ,illustrate CPU1 There is sequence variation in itself, which leads to phenotypic variation in the phosphorus uptake efficiency of soybean populations. Therefore, finding this functional variation site will provide valuable information for later gene editing breeding and precision molecular marker-assisted selection breeding.

[0051] Based on the resequencing results and genome-wide association analysis results in Example 1, the inventors found that there are mainly two CPU1 alleles of CPU1 - H1 (The nucleotide sequence is shown in SEQ ID NO.5) and CPU1- H2 (The nucleotide sequence is shown in SEQ ID NO.1), and the variation site significantly associated with phosphorus uptake efficiency exists in the promoter region and 5'UTR, and no associated signal is found in the coding region, implying the variation of phosphorus uptake efficienc...

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Abstract

The invention discloses an SEC12-like protein gene CPU1 and application thereof in improving soybean phosphorus efficiency, and belongs to the field of biotechnology. Through a forward genetics method of whole genome association analysis, a major genetic locus influencing soybean phosphorus absorption efficiency is identified, and a candidate gene CPU1 therein is identified. The gene CPU1 has natural variation in soybeans, and comprises two alleles, namely a phosphorus low-efficiency allele CPU1-H1 and a phosphorus high-efficiency allele CPU1-H2. The research based on the whole transformed plant of the soybean shows that the phosphorus absorption efficiency of the soybean is remarkably reduced by inhibiting the expression of the allele CPU1-H2, and the biomass and yield of the transgenic plant are finally reduced. According to the invention, a new scientific insight is provided for genetic basic research of natural variation of crops including soybeans, and meanwhile, an excellent allele resource is provided for phosphorus high-efficiency molecular breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to SEC12-like protein gene CPU1 and its application in improving phosphorus efficiency. Background technique [0002] Soybean, as an important grain, oil and feed crop in my country, provides abundant protein and oil. Although my country is the origin of soybeans, it has been the world's largest producer, consumer and exporter of soybeans for a long time; but since 1996, my country has become a net importer of soybeans and needs to import a large amount of soybeans from the Americas every year. Soybeans have serious hidden dangers in food security (Shi Hui et al., 2018). At the same time, soybean, as the leguminous crop with the largest amount of biological nitrogen fixation, has outstanding performance in reducing weight and increasing efficiency and reducing environmental pollution (Xinxin Li et al., 2016). Therefore, improving my country's soybean production capacity is of great si...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/06A01H6/54
CPCC07K14/415C12N15/8261C12N15/8242C12N15/8205Y02A40/146C12N15/52A01H5/10A01H6/542C12N15/8273C12N15/8271
Inventor 郭子龙陈志长廖红
Owner FUJIAN AGRI & FORESTRY UNIV
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