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Malic enzyme gene SgME1 as well as application thereof

A malic enzyme gene and malic enzyme technology, applied in the field of plant biology, can solve problems such as aluminum stress

Inactive Publication Date: 2014-01-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]Through the early screening of aluminum tolerance, we found that the phosphorus-inefficient Stylo genotype Fine-stem is sensitive to aluminum stress, and the phosphorus-efficient Stylo genotype TPRC2001-1 is sensitive to aluminum stress. Aluminum toxicity has good tolerance (Du et al., 2009), and the aluminum tolerance of TPRC2001-1 is comparable to that of rice, which is an aluminum-tolerant crop.

Method used

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  • Malic enzyme gene SgME1 as well as application thereof
  • Malic enzyme gene SgME1 as well as application thereof
  • Malic enzyme gene SgME1 as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] SgME1 gene cloning

[0052] 1. Isolation and identification of SgME1 protein

[0053] Refer to the method of Chen Zhijian et al. (2011) to extract the root protein of Stylophyllum, and perform two-dimensional electrophoresis. Three replicates were set up for each experimental treatment, and the protein spots with significant differential expression between the control without aluminum and the treatment with aluminum were screened, and ABI 4700 TOF-TOF (Applied Biosystems, USA) was used for MALDI-TOF / TOF MS mass spectrometry analysis.

[0054] The result is as Figure 4 As shown, it can be seen from the results that: compared with the control, the accumulation of SgME1 protein in the roots of Stylophyllum 24 hours after aluminum stress has no significant change. However, after 72 hours of Al stress, the accumulation of SgME1 protein was significantly increased in roots, indicating that the expression of SgME1 protein was enhanced by long-term Al stress.

[0055] 2. ...

Embodiment 2

[0087] Research on genetically modified materials

[0088] 1. Acquisition of genetically modified material

[0089] (1) Acquisition of yeast transgenic strains

[0090] Referring to SC easy comp transformation kit (Invitrogen, USA), pYES2- SgME1 Yeast INVSC1 was transformed with pYES2 empty vector, respectively.

[0091] (2) Obtaining the hairy root of transgenic kidney bean

[0092] The constructed expression vector was transformed into Agrobacterium rhizogenes K599, and the transgenic hairy roots were obtained by Agrobacterium-mediated induction of kidney bean hairy roots in vitro.

[0093] (3) Obtaining transgenic Arabidopsis plants

[0094] The constructed expression vector plasmid was transformed into Agrobacterium tumefaciens Gv3101, and transgenic Arabidopsis plants were obtained by Agrobacterium-mediated Arabidopsis inflorescence transfection method.

[0095] 2. Detection of genetically modified materials

[0096] (1) Detection of yeast transgenic strains

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PUM

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Abstract

The invention discloses a gene SgME1 which codes malic enzyme as well as an application of the gene SgME1. The nucleotide sequence of the malic enzyme gene SgME1 is shown as SEQIDNO:1, and the amino acid sequence of coded proteins is shown as SEQIDNO:2. The SgME1 gene has the functions of regulating and controlling synthesis of malic acid of stylosanthes guianensis, and over-expressing the SgME1 to accelerate synthesis of malic acid of yeasts, hairy roots of beans and Arabidopsis and increasing the aluminum tolerance of the SgME1.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to malic enzyme gene SgME1 and its application. Background technique [0002] Aluminum toxicity is one of the major obstacles limiting crop growth on acidic soils (Kochian et al., 2004). When the pH is less than 5, aluminum in the soil exists in an active form, which can be quickly absorbed by plant roots (Kinraide, 1991), and has a toxic effect on the growth of root tips, which eventually leads to a decrease in crop yield. [0003] Aluminum ions can interact with cell walls and symplasts, thereby disturbing the normal physiological and biochemical reactions of plant cells (Kochian et al., 2005). Studies have shown that aluminum easily binds to the negatively charged pectin of the cell wall, thereby reducing the extensibility of the cell wall and limiting cell elongation (Yang et al., 2011). In addition to combining with the cell wall, aluminum can also accumulate in the sympla...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/04C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/82A01H5/06A01H5/00
Inventor 田江廖红孙丽莉
Owner SOUTH CHINA AGRI UNIV
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