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Application of paddy OsPEX1 gene in regulating lignin metabolism

A technology of metabolic regulation and lignin, applied in the application field of rice extensin gene OsPEX1, can solve the problems of few research reports, and achieve the effect of broadening the genetic basis, improving the lodging resistance and enriching the molecular mechanism.

Active Publication Date: 2019-04-05
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there have been relatively in-depth studies on lignin synthesis and metabolism, the research objects are mainly concentrated on plants such as Arabidopsis thaliana, Nicotiana tabacum, Medicago Sativa, and Zeamays. There are still few research reports on the regulation of lignin synthesis metabolism

Method used

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  • Application of paddy OsPEX1 gene in regulating lignin metabolism
  • Application of paddy OsPEX1 gene in regulating lignin metabolism
  • Application of paddy OsPEX1 gene in regulating lignin metabolism

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1 Phenotype identification of OsPEX1 mutants

[0037] 1. Cytological observation of OsPEX1 mutants

[0038] In the present invention, the wild-type Zhonghua 11 and the third section of the stem node of the OsPEX1 mutant are cross-sectioned freehand, and the cytology is observed by a fluorescence microscope under an ultraviolet light environment. The results showed that compared with the wild type, pex1 heterozygous mutants and homozygous mutants had significantly enhanced lignin autofluorescence and thickened cell walls, and the trend was pex1 homozygous mutant > pex1 heterozygous mutant > wild type, In addition, the xylem cells of OsPEX1 mutants were also relatively smaller and more closely arranged ( figure 1 (a)). These results suggest that the increased lignin content in the cell wall of OsPEX1 mutants may cause cell wall thickening and limit the normal loose growth of cells, resulting in smaller cell morphology of OsPEX1 mutants. The above results indica...

Embodiment 2

[0111] Example 2 Antisense suppression vector transformation verification OsPEX1 gene function

[0112] In order to further verify the regulatory effect of the OsPEX1 gene on rice lignin metabolism, this example also constructed the antisense suppression (antisense suppression) vector pRNAi-OsPEX1 of the OsPEX1 gene, and introduced it into Zhonghua 11 (referred to as ZH11), and observe and analyze the phenotype of the transgenic plants. Specific steps are as follows:

[0113] 2.1 Construction of antisense suppression vector pRNAi-OsPEX1

[0114] According to the sequence structure of the OsPEX1 gene, a pair of RNAi primers OsPEX1-RNAiF / OsPEX1-RNAiR with restriction sites were designed. The primer sequences are as follows:

[0115] OsPEX1-RNAiF: 5′- GGATCC AGGAGTCACCTGAGGAACCA-3' (the underline is the BamHI restriction site);

[0116] OsPEX1-RNAiR: 5′- CTGCAG AAGCTTATTCTTCTGGAGTCGGTGGA-3' (the underline is the restriction site of PstI).

[0117] Extract the total RNA of...

Embodiment 3

[0127] Example 3 OsPEX1 gene expression is positively correlated with lignin content

[0128] The stem of wild-type rice ZH11 was divided into 5 segments, and lignin-specific staining and content determination were performed on each segment, and the relative expression of OsPEX1 gene in each segment was analyzed. Wherein, the specific steps of lignin staining and content determination and OsPEX1 gene expression analysis refer to Example 2, and the primer sequences used are: RT-OsPEX1F: 5′-GAGATGGGCTACCTGCAGAACA-3′; RT-OsPEX1R: 5′-GTAGGCGAAGCTGAAGTTGACG-3 '. The results showed that in different segments of the same stem node, as the expression of OsPEX1 gene increased, the lignin content also increased, and the expression of lignin synthesis-related genes was up-regulated ( image 3 ).

[0129] Based on the above results, the phenotype of high lignin content in OsPEX1 mutants is caused by the high expression of OsPEX1 gene, and down-regulating the expression of OsPEX1 will re...

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Abstract

The invention discloses application of a paddy OsPEX1 gene in regulating lignin metabolism and belongs to the field of plant genetic engineering. The OsPEX1 gene disclosed by the invention has a DNA sequence shown as SEQ ID NO:1; phenotypic analysis for mutants and analysis for antisense suppression of transgenic progeny strains prove that the OsPEX1 gene plays an important role in regulating lignin metabolism; the cloning of OsPEX1 gene is capable of enriching a molecular mechanism for regulating the lignin metabolism of paddy, supplying a new gene resource to lodging-resistant genetic improvement of paddy, and supplying a new material choice for molecular design and breeding of paddy; the paddy variety with increased lignin content and improved lodging resistance can be bred in the manner of increasing the OsPEX1 expression in paddy; operability is ultrahigh; a new path is established for broadening the genetic basis of present lodging-resistant variety.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to the application of a rice extensin gene OsPEX1. The invention studies and verifies the biological function of a rice OsPEX1 gene. The invention also relates to the application method of using the carrier to transform plant cells to change rice lignin content and obtain rice with strong lodging resistance. Background technique [0002] Lignin is one of the important components of plant cell walls, second only to cellulose, and is the main substance formed in the process of secondary cell wall thickening in vascular plants. Lignin is closely related to the growth and differentiation of plants and the development of cell walls. During the lignification and metabolism of plants, lignin is mainly accumulated in the fiber cells between vessel cell walls and vascular bundles, which can enhance the hardness of plant cell walls, mechanical support and resistance. Com...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46A01H1/02
CPCA01H1/02C07K14/415C12N15/8255C12N15/8261
Inventor 张向前柯善文栾鑫钟天秀陈曙解新明刘树春代航
Owner SOUTH CHINA AGRI UNIV
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