Lycoris chinensis LcCLA gene VIGS silencing vector, silencing system and construction method and application thereof

A construction method, silent technology, applied in application, horticultural methods, genetic engineering, etc., can solve the problems of unestablished and applied

Pending Publication Date: 2022-04-19
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has been applied to a variety of plants for gene function verification, but this system has not been established and applied in Lycoris in China

Method used

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  • Lycoris chinensis LcCLA gene VIGS silencing vector, silencing system and construction method and application thereof
  • Lycoris chinensis LcCLA gene VIGS silencing vector, silencing system and construction method and application thereof
  • Lycoris chinensis LcCLA gene VIGS silencing vector, silencing system and construction method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Cloning of the specific fragment of LcCLA gene

[0044] According to the sequence assembly, annotation and comparison analysis of the Chinese Lycoris transcriptome, it was found that LcCLA has a complete ORF (full length 1980bp). The total RNA of Amaryllis lycoides leaves was extracted with an RNA extraction kit (purchased from Beijing Huayueyang Biotechnology Co., Ltd., catalog number 0416-50gk), and after reverse transcription with oligo(dT)18 (purchased from Thermo Fisher, catalog number SO131), Design full-length primers CLA1-F (denoted as SEQ ID NO.2) according to the Chinese Lycoris transcriptome: 5'-TTGACGGGCAGGAGGGAC-3' and CLA1-R (denoted as SEQ ID NO.3): 5'-GCGGAGGAAGCAGTTTAGGC-3 ', LcCLA-specific bands were obtained by PCR amplification, agarose gel electrophoresis, gel cutting purification, cloning and sequencing were performed to obtain the final sequence.

[0045] (2) Vector construction

[0046] According to the final sequence design of step (1), pr...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a lycoris chinensis LcCLA gene VIGS silence vector, a silence system and a construction method and application of the silence system. According to the invention, a Chinese lycoris radiata LcCLA gene VIGS silencing vector and a silencing system are constructed, the silencing system is introduced into Chinese lycoris radiata leaves by adopting a needle injection mode to induce silencing of Chinese lycoris radiata endogenous LcCLA, the expression level of the Chinese lycoris radiata LcCLA gene is effectively reduced, a virus-induced gene silencing plant is obtained, and the function of the Chinese lycoris radiata LcCLA gene can be verified; the application value is realized in the functional gene research of the lycoris chinensis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Chinese Lycoris LcCLA gene VIGS silencing vector, a silencing system, a construction method and application thereof. Background technique [0002] Lycoris chinensis Traub is an important ornamental plant of the Lycoris genus. It has ornamental characteristics such as bright colors, beautiful flower shapes, tall and straight flower stems, etc., and has low environmental requirements and strong adaptability. It is a good choice for landscaping . Molecular breeding is the trend and common method of flower breeding research in the future. It can create excellent conditions for improving ornamental traits, improving flowering period, and cultivating new germplasm. However, the genetic transformation system of Lycoris plants is not yet mature, and it is difficult to study gene functions. [0003] Virus-induced gene silencing (VIGS) is a technique based on post-transcriptiona...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/83C12N15/84C12N15/66C12N15/29A01H6/04A01H5/12A01G7/06
CPCC12N15/8218C12N15/8205C07K14/415C12N15/66A01G7/06
Inventor 张凤姣程光昊王忠王芝权束晓春王宁王涛
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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