Application of important gene GmSWEET6 of soybean sucrose transporter

A sucrose transporter, soybean technology, applied in the field of plant genetic engineering and biology, can solve the problem of no SWEET transporter research

Active Publication Date: 2018-08-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, in mycorrhizal plants, no mycorrhizal-induced SWEET transporter responsible for efflux of sucrose to the symbiotic interface has been found, and there is no related study on mycorrhizal-induced SWEET transporter in soybean.

Method used

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  • Application of important gene GmSWEET6 of soybean sucrose transporter
  • Application of important gene GmSWEET6 of soybean sucrose transporter
  • Application of important gene GmSWEET6 of soybean sucrose transporter

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Embodiment 1

[0039] Analysis of the expression pattern of the GmSWEETs gene family in soybean roots: The soybean SWEET gene family has a total of 53 members (Patil et al., 2015). According to the evolutionary homologous relationship, the third branch related to sucrose transport and microbial symbiosis was screened28 Then, quantitative PCR technique was used to determine the expression of GmSWEETs family member genes under different soybean varieties, different inoculation treatments and different phosphorus levels.

[0040] Using two soybean varieties, phosphorus-efficient Brazilian 10 (BX10) and phosphorus-inefficient Native 2 (BD2), two AMF inoculation treatments were set up: no inoculation (NM) and inoculation with AM fungus Rhizophagus irregularis (Ri). Set up two phosphorus treatments, 50 μM KH 2 PO 4 As low phosphorus (LP) and 500 μM KH 2 PO 4 K as High Phosphorus (HP), Low Phosphorus Treatment + with K 2 SO 4 make up. Use 1 / 2 Hoagland nutrient solution for sand cultivation. ...

Embodiment 2

[0055] 1. Cloning of GmSWEET6 gene: using the soybean YC03-3 leaf cDNA as a template, using the upstream specific primer 5′-ATCG CCCGGG ATGTCGTCCCACAGTCATTCTAA-3' (SEQ ID NO: 5) and downstream specific primer 5'-ATCG CCCCGGG TCAAACTTCGCAACTGATCACCC-3' (SEQ ID NO: 6) amplified the full-length ORF sequence of 864bp of the GmSWEET6 gene, and sequenced and compared the obtained GmSWEET6 coding sequence, see Glyma.04G198600, and the corresponding protein sequence, see Glyma.04G198600.

[0056] 2. GmSWEET6 gene promoter cloning, vector construction and tissue expression localization analysis: Promoter analysis and expression vector construction: according to conventional methods, extract genomic DNA from leaves of soybean YC03-3 genotype, use soybean leaf genomic DNA as a template, and use Upstream specific primer 5′-CTATGACATGATTAC GAATTC CCACCTTGTTATACCTCATT-3' (SEQ IDNO: 7) and downstream specific primer 5'-GACTGACCTACCCGG GGATCC GGAATTTCTCTCTCTCCTCT-3′(SEQ IDNO:8) amplifie...

Embodiment 3

[0059] Construction of subcellular localization experiment vector: using soybean YC03-3 leaf cDNA as template, using upstream specific primer 5'-GGGGacaagtttgtacaaaaaagcaggcttcATGTCGTCCCACAGTCATTCTAA-3' (SEQ ID NO: 9) and downstream specific primer 5'-GGGGaccactttgtacaagaaagctgggtcTCAAACTTCGCAACTGATCACCC-3' (SEQ ID NO: 10) Amplify the full-length ORF sequence of 864bp of the GmSWEET6 gene, recover the fragment by PCR, and connect the GmSWEET6 gene to the destination vector pMDC43 by Gatway technology. Escherichia coli DH10B was transformed, and Agrobacterium GV3101 was transformed after the sequence was correct for subcellular localization experiments.

[0060] For the subcellular localization analysis of GmSWEET6, the method of transient transformation of tobacco epidermal cells was used. Shake the GV3101 bacterial liquid fused with the GmSWEET6 gene and the GV3101 bacterial liquid transformed into the membrane Marker plasmid 1008 overnight, then centrifuge, and use 2 And th...

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Abstract

The invention discloses application of an important gene GmSWEET6 of a soybean sucrose transporter and relates to the fields of plant genetic engineering and biotechnologies. According to a real-timefluorescent quantitative PCR (Polymerase Chain Reaction) method, the sucrose transporter gene GmSWEET6 with mycorrhiza induced expression is identified in soybeans. The expression of the gene is regulated by mycorrhiza fungus infection. Under mycorrhiza fungus inoculation conditions, over-expressed or interference expressed GmSWEET6 in transgenic soybean plants has an effect of regulating plant growth and phosphorus absorption and has a significant influence on beneficial symbiosis of the soybeans and arbuscular mycorrhiza fungi, which has significance for explaining biological functions of the SWEET genes in symbiosis of leguminous crops and the mycorrhiza fungi so as to regulate beneficial symbiosis between plants and the mycorrhiza fungi.

Description

technical field [0001] The invention relates to the fields of plant genetic engineering and biotechnology, in particular to the application of an important gene GmSWEET6 of soybean sucrose transporter. Background technique [0002] Higher plants are autotrophic organisms composed of autotrophic and heterotrophic organs. Heterotrophic organs require photosynthetic products from autotrophic organs for growth and development. 80% of photosynthetic compounds synthesized in "source" organs such as mature leaves of plants are transported to "sink" organs such as roots, flowers and fruits through plant vascular system. Sucrose is one of the photosynthetic compounds of green plants, and it is also the main form of transport and distribution of photosynthetic products in most higher plants (Yang Shuang et al., 2005; Bai Xuemei et al., 2006). The directional transport and distribution of sucrose in plants not only regulates the entire growth and development process of plants, but al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/54
CPCC07K14/415C12N15/8261
Inventor 王秀荣陈阿赵少鹏陈康
Owner SOUTH CHINA AGRI UNIV
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