VIGS silence system for identifying peony UDPglucose Flavonoid Glucosyl-Transferase gene

A flavonoid glycosyl and transferase technology, which is applied in genetic engineering, plant genetic improvement, and the introduction of foreign genetic material using vectors, etc., can solve the problems of peony regeneration and genetic transformation, and flowering-related gene phenotype identification difficulties, etc.

Active Publication Date: 2017-10-13
INST OF BOTANY CHINESE ACAD OF SCI
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Problems solved by technology

[0004] Peony regeneration and genetic transformation are very difficult, there is no report on its genetic transformation system, and there is a technical bottleneck in the verification of the function of peony genes using transgenic technology
In addition, peony takes about 3 years from seedling to flowering, and it is difficult to identify the phenotype of flowering-related genes

Method used

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  • VIGS silence system for identifying peony UDPglucose Flavonoid Glucosyl-Transferase gene
  • VIGS silence system for identifying peony UDPglucose Flavonoid Glucosyl-Transferase gene
  • VIGS silence system for identifying peony UDPglucose Flavonoid Glucosyl-Transferase gene

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Embodiment 1

[0043] 1. Gene cloning

[0044] The extraction of total RNA from petals was performed according to the kit instructions (RNAprep Pure Plant Kit, FastQuantcDNA Tiangen Biochemical Technology (Beijing) Co., Ltd.). According to the UF3GT and UF5GT sequences in public databases, the upstream and downstream primers are designed to clone the target fragment. Design the upstream and downstream primers for the PsUF3GT and PsUF5GT fragments for VIGS silencing and add BamH I and Xho I restriction sites. Amplification primers are shown in Table 1.

[0045] Table 1 Sequence related information of primers used in the present invention

[0046]

[0047]

[0048] Note: The underline indicates the base restriction site.

[0049] (1) Cloning sequence analysis of PsUF3GT target fragment

[0050] The length of the PsUF3GT target fragment amplified by the upstream and downstream primers is 413 bp, and its nucleotide sequence is shown in sequence 1 in the sequence table. The PCR reaction system and proc...

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Abstract

The invention discloses a VIGS silence system for identifying a peony UDPglucose Flavonoid Glucosyl-Transferase gene. The VIGS silence system for identifying a peony UDPglucose Flavonoid Glucosyl-Transferase gene comprises a recombinant vector which is obtained by adding desired genes to a VIGS silence vector. The desired genes are nucleotide sequences shown as a first sequence in a sequence table and / and nucleotide sequences shown as a third sequence in the sequence table. The UDPglucose Flavonoid Glucosyl-Transferase gene PsUF3GT and a PsUF5GT conserved segment are cloned by homology-based cloning technologies as target genes to construct TRV2 vector, so that the VIGS silence system is built successfully, and amount of gene expression of the UDPglucose Flavonoid Glucosyl-Transferase gene PsUF3GT and the PsUF5GT conserved segment and total anthocyanin content are reduced. The system has great significance for peony genes function study and lays foundation for analyzing an molecular mechanism formed by peony splash.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a VIGS silencing system for identifying peony flavonoid glycosyltransferase genes. Background technique [0002] The peony (Paeonia suffruticosa) is a deciduous subshrub of the Paeoniaceae (Paeoniaceae) and the Section Moutan of the peony group (Paeonia). It is a traditional famous flower in my country. The research on the chemical basis of flower color formation of peony is more in-depth. The petals contain 12 kinds of anthocyanins, namely pelargonidin 3-glucoside (Pg3G), pelargonidin 3,5-diglucoside (Pg3G5G), and cyanidin 3-glucoside (Cy3G), cyanidin 3,5-diglucoside (Cy3G5G), paeoniflorin 3-glucoside (Pn3G), paeoniflorin 3,5-diglucoside (Pn3G5G), cyanidin 3, 5-Dihexoside (Cy-3,5-di-hexoside), cyanidin 3-glucose-5-arabinoside (Cy3G5Ara), cyanidin 3-arabinoside (Cy3Ara), cyanidin 3 -Galloyl glucoside (Cy3galloylG); Paeoniflorin 3-arabinosides (Pn3Ara) and Paeoniflorin 5-galloyl ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/83C12N15/66C12N15/54
Inventor 舒庆艳王亮生朱瑾
Owner INST OF BOTANY CHINESE ACAD OF SCI
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