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Malic acid transporter gene GmALMT1 and application thereof

A technology of malic acid and transgenic plants, applied in the field of plant biology, can solve the problems that key genes have not been reported yet

Active Publication Date: 2013-02-13
嘉兴卓十生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have found that soybean genotypes adapted to southern acidic soils have higher phosphorus efficiency and aluminum toxicity tolerance, but the key genes controlling the process have not been reported

Method used

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  • Malic acid transporter gene GmALMT1 and application thereof
  • Malic acid transporter gene GmALMT1 and application thereof
  • Malic acid transporter gene GmALMT1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] GmALMT1 gene cloning

[0053] 1. Gene ( GmALMT1 ) clone

[0054] According to the homology comparison of the reported amino acid sequences of wheat TaALMT1-1 (AB081803) and TaALMT1-2 (AB081804), Arabidopsis AtALMT1 (At1g08430) and rapeseed BnAlMT1 (AB194300) BnAlMT2 (AB194301) genes, the highly conserved regions Design the upstream degenerate primer 5'-TGGGCIRTIHTIACIGTIGT-3'(SEQ ID NO:3) and the downstream degenerate primer 5'-CCIGCCCAIAYIGGRMA-3'(SEQ ID NO:4), and extract the phosphorus efficient genotype according to the TRIzol one-step method HN98 soybean root tip total RNA, and then the cDNA obtained by reverse transcription was used as a template for PCR amplification. The PCR reaction system was 50 μL, including 1 μL of 10 μM forward and reverse primers, 5 μL of 10×PCR buffer, and 2.5 mM dNTP 4 μL, Taq enzyme 0.5 μL, cDNA template volume 2.5 μL, then sterilized ddH 2 O to make up 50 μL. The PCR reaction program was: 1 minute at 94°C; 30 cycles of amplifi...

Embodiment 2

[0098] Example 2 Research on Transgenic Compound Plants

[0099] 1. Obtaining genetically modified materials

[0100] 1) Obtaining transgenic soybean compound plants

[0101] The constructed expression vector and interference vector were transformed into Agrobacterium rhizogenes K599, and the transgenic composite plants (transgenic hairy roots in the root and non-transgenic in the shoot) were obtained by Agrobacterium-mediated soybean hypocotyl injection. All phenotypes were identified using this strain. Empty vector control: According to the above method, the expression vector pYLRNAi was transformed into soybean in the same way to obtain a pYLRNAi-transformed empty vector control line (CK).

[0102] 2) Obtaining transgenic Arabidopsis plants

[0103] The constructed expression vector plasmid was transformed into Agrobacterium tumefaciens Gv3101, and transgenic Arabidopsis plants were obtained by Agrobacterium-mediated Arabidopsis inflorescence transfection method. Empt...

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Abstract

The invention discloses a malic acid transporter gene GmALMT1 and an application thereof. The nucleotide sequence of the malic acid transporter gene GmALMT1 is shown as SEQ ID NO.1. An amino acid sequence of an encoded protein is shown as SEQ ID NO.2. The GmALMT1 gene provided by the invention can be used for regulating soybean root tip malic acid secretion.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to a malate transporter gene GmALMT1 and its application. Background technique [0002] Acidic soils are widely distributed in the world. The total area of ​​acidic soils in the world accounts for 30-40% of the world's cultivated land, and more than 50% of the potential cultivated land is acidic soils, mainly distributed in tropical, subtropical and temperate regions (Kochian et al. , 2004). The effect of acidic soil on crop growth inhibition is not only manifested in H + Excessive concentrations are harmful to the crop itself, and often appear as a synergistic inhibition of plant growth and yield by the lack of available phosphorus and aluminum toxicity. [0003] During the long-term evolutionary process of natural and artificial selection, plants have formed a series of morphological, physiological and molecular cooperative adaptive mechanisms to overcome obstacles such as lo...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C07K14/415A01H5/00
Inventor 梁翠月田江廖红
Owner 嘉兴卓十生物科技有限公司
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