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A rate-limiting gene for brassinolide synthesis regulating poplar wood formation and its application

A technology of brassinosteroids and genes, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as accelerating forest tree breeding, and achieve the effect of increasing the number of cells

Active Publication Date: 2021-01-26
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

[0005] At present, the role of brassinosteroids in woody plants has not been reported. Therefore, using woody plants as research objects, combined with molecular biology and genetic engineering techniques, to analyze the molecular mechanism of brassinosteroids regulating the development of poplar wood. It is of great significance to understand the molecular basis of root development of woody plants, as well as improve difficult-to-root forest trees through molecular breeding and accelerate forest tree reproduction

Method used

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  • A rate-limiting gene for brassinolide synthesis regulating poplar wood formation and its application
  • A rate-limiting gene for brassinolide synthesis regulating poplar wood formation and its application
  • A rate-limiting gene for brassinolide synthesis regulating poplar wood formation and its application

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Embodiment 1

[0034] Step (1): Cloning the PagDET2 gene

[0035] Using 84K (P.alba X P.glandulosa) Populus alba as material, use RNeasy Plant Mini kit and RNase-free DNase I kit (Qiagen, Hilden, Germany) to extract the total RNA of tissue culture plantlet 84K; each sample About 2.0 μg of RNA was synthesized by using SuperScript III first-strand synthesis system (LifeTechnologies, Carlsbad, CA, USA) to synthesize the first strand of cDNA; refer to the published Populus trichocarpa genome sequence, and use Primer3 software to design primers (the amplicon contains the initial codon and stop codon) for full-length amplification of the gene (GATEWAY linker is introduced into the primer);

[0036] Wherein, the PagDET2 CDS forward primer is (such as sequence 4 in the sequence listing), see Table 1 below;

[0037] Table 1

[0038]

[0039] The PagDET2 CDS reverse primer is (such as sequence 5 in the sequence listing) see Table 2 below;

[0040] Table 2

[0041]

[0042] The high-fidelity PC...

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Abstract

The invention relates to a rate-limiting gene for brassinosteroid synthesis that regulates poplar wood formation and its application, and belongs to the technical field of plant genetic engineering; its nucleotide sequence is shown in sequence 1 in the sequence table, and its amino acid sequence is shown in sequence Shown in sequence 2 in the list. The present invention transfers the PagDET2 gene into poplar, and the transgenic poplar overexpressing PagDET2 is significantly taller and thicker than the wild type, and the number of xylem cells is significantly increased, indicating that the PagDET2 gene is a key regulatory gene regulating the formation of poplar wood. It has important application value in the field of genetic engineering of forest trees and the field of clonal forestry.

Description

technical field [0001] The invention relates to a brassinosteroid synthesis gene for regulating the formation of poplar wood and its application, in particular to PagDET2, a rate-limiting gene for the synthesis of brassinosteroid for regulating the formation of poplar wood, and its application, which belongs to the technical field of biological genetic engineering. Background technique [0002] Wood is the most important renewable resource on earth and the main raw material for pulp and paper. It is an important way to meet the market demand by genetically improving the existing main afforestation tree species to increase wood production. Poplar itself is an economic tree species widely distributed in the world. It has the characteristics of fast-growing early, strong adaptability, wide distribution, many varieties, easy hybridization, easy improvement of heredity, and easy reproduction, so it is widely used in intensive cultivation. Due to its excellent characteristics, po...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/82C12N15/64A01H6/00
CPCC12N9/0004C12N15/64C12N15/8261C12N15/8298
Inventor 杜娟李泽华刘颖丽卢孟柱
Owner ZHEJIANG UNIV
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