Brassinosteroid synthesizing rate-limiting gene for adjusting and controlling poplar wood formation and application of brassinosteroid synthesizing rate-limiting gene

A Brassinolide, gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as accelerating forest tree breeding, and achieve the effect of increasing the number of cells

Active Publication Date: 2019-07-30
ZHEJIANG UNIV
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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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  • Brassinosteroid synthesizing rate-limiting gene for adjusting and controlling poplar wood formation and application of brassinosteroid synthesizing rate-limiting gene
  • Brassinosteroid synthesizing rate-limiting gene for adjusting and controlling poplar wood formation and application of brassinosteroid synthesizing rate-limiting gene
  • Brassinosteroid synthesizing rate-limiting gene for adjusting and controlling poplar wood formation and application of brassinosteroid synthesizing rate-limiting gene

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Embodiment 1

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

[0035] Using 84K (P.alba X P.glandulosa) poplar as material, RNeasy Plant Mini kit and RNase-free DNase I kit (Qiagen, Hilden, Germany) were used to extract the total RNA of tissue culture seedlings 84K; each sample About 2.0 μg of RNA was taken to synthesize the first strand of cDNA by using the SuperScript III first-strand synthesis system (Life Technologies, Carlsbad, CA, USA); primers were designed using Primer3 software with reference to the published P. codons and stop codons), perform full-length gene amplification (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 PCR reaction system is as follows: TaKaRa high-fidelity amp...

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Abstract

The invention relates to a brassinosteroid synthesizing rate-limiting gene for adjusting and controlling poplar wood formation and application of the brassinosteroid synthesizing rate-limiting gene, and belongs to the technical field of plant gene engineering. The nucleotide sequence of the brassinosteroid synthesizing rate-limiting gene is shown in a sequence 1 in a sequence table, and the aminoacid sequence of the brassinosteroid synthesizing rate-limiting gene is shown in a sequence 2 in the sequence table. The PagDET2 gene is transferred into a poplar tree, compared with wild-type polar tree, the transgenic polar tree with the PagDET2 of overexpression significantly grows tall and becomes thick, in addition, the cell number of the xylem of the transgenic polar tree is significantly increased, it is proved that the PagDET gene is a key adjusting gene for adjusting and controlling poplar wood formation, and the brassinosteroid synthesizing rate-limiting gene has important application value in the fields of forest-tree gene engineering and clonal forestry.

Description

technical field [0001] The invention relates to a brassinolide synthesis gene that regulates the formation of poplar wood and its application, in particular to a Brassinolide synthesis rate-limiting gene PagDET2 that regulates the formation of poplar wood and its application, belonging 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 genetic improvement of the existing main afforestation tree species to increase the wood yield. Poplar itself is an economic tree species widely distributed in the world. It has the characteristics of early growth, strong adaptability, wide distribution, many varieties, easy hybridization, easy improvement of heredity, and easy reproduction. Therefore, it is widely used in intensive cultivation. With its excellent characteristics, poplar has always been an...

Claims

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Application Information

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Patent Type & Authority Applications(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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