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Application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom

A technology of glycosyltransferase and lignin, which is applied in the field of genetic engineering to achieve the effect of increasing lignin content and satisfying industrial and agricultural production

Inactive Publication Date: 2012-07-04
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After retrieval, the gene of poplar glycosyltransferase family 1 PtGT1 There is no report on the application of increasing plant lignin content and promoting flowering

Method used

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  • Application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom
  • Application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom
  • Application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom

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Experimental program
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Effect test

Embodiment 1

[0018] 1. Poplar Glycosyltransferase Gene PtGT1 clone

[0019] Take poplar test-tube seedlings grown on MS medium for 2-3 weeks, use TRIzol kit to extract RNA, and RT-PCR method to amplify the full length PtGT1 Gene, to obtain its cDNA sequence (1446bp), the amplification primers used are:

[0020] GT1-a: 5'-ATAGGATCCATGCAAAACACAAAACCTCA-3';

[0021] GT1-b: 5'-ATACCCGGGCTAGGCACCTTGAGCCTTTG-3';

[0022] The amplified product is recovered for later use.

[0023] 2. Poplar Glycosyltransferase Gene PtGT1 Sequence information and characteristic analysis of

[0024] PtGT1 The cDNA of the coding region of the gene is 1446bp, encoding a protein consisting of 481 amino acids, and the molecular weight of the protein is 66.4kDa. The gene has no introns and is a continuous coding gene. Sequence analysis found that there is a PSPG conserved sequence consisting of 44 amino acids near the C-terminus of the protein, which is a common feature of plant secondary metabolite glycosyltrans...

Embodiment 2

[0030] 1. contains PtGT1 Construction of cDNA expression vector for coding region

[0031] with plant expression vector restriction sites PtGT1 blunt end ligation Eco The pBluescriptSK plasmid was single-cut by RV to form the intermediate vector pSKGT1.

[0032] will sequence the correct PtGT1 gene pass Bam HI and Sma I was excised from the intermediate vector pSKGT1. then with Sac I digestion, fill-in and Bam The HI digested plant expression vector pBI121 was ligated to form a gene containing the target gene PtGT1 The plant expression vector pBIGT1.

[0033] 2. Agrobacterium-mediated plant transformation

[0034] Pick a single colony of Agrobacterium containing vector pBIGT1 in YEB medium containing antibiotics: 50mg / L Kanamycin (Kan), 50mg / L Rifampicin (Rif), culture in a shaker at 28°C, 200rpm for 48h Left and right; After the bacteria are shaken up, transfer 2ml of the bacteria to 50ml of YEB medium, culture in a shaker at 28°C at 200rpm. When the cell OD ...

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Abstract

The invention discloses application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom. A nucleotide sequence of the glycosyl transferase gene PtGT1 is shown in an SEQIDNo.1 and is cloned from Chinese white poplar through RT-PCR technology. The application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom utilizes gene PtGT1 to construct plant over expression vector to perform plant transgene operation to obtain transgene plant. Detection shows that the lignin content of transgene plants is remarkably increased, and simultaneously plants are promoted to blossom earlier. Thus, by means of the application of poplar glycosyl transferase gene PtGT1 in improving plant lignin content and promoting blossom, novel plants can be created, and requirements of industrial and agricultural production can be well met.

Description

[0001] technical field [0002] The present invention relates to the application of a glycosyltransferase gene, in particular to a poplar glycosyltransferase gene PtGT1 The application in increasing plant lignin content and promoting flowering belongs to the field of genetic engineering. Background technique [0003] Glycosyltransferase is an enzyme specifically responsible for catalyzing the glycosylation modification of plant molecules. It transfers active sugars from nucleoside sugars, usually from uridine nucleoside diphosphate-glucose, to hormones, secondary metabolites, On the receptors of a series of plant small molecule compounds such as pathogen infectants and exogenous toxic substances in plants. Glycosylation will change the biological activity, water solubility, stability, transport characteristics in cells and in the whole plant, subcellular localization, and mutual recognition and binding characteristics with receptors of plant small molecule compounds, and ca...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 侯丙凯王艳文
Owner SHANDONG UNIV
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