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A gene for regulating the formation of tea leaves and its application

A technology of tea hair and gene, which is applied in the field of tea tree MYB transcription factor gene and its encoded protein, can solve the problems of unclear molecular mechanism, hinder the in-depth understanding of tea tree development biology and tea quality formation mechanism, and achieve rich understanding know the effect

Active Publication Date: 2020-11-03
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tea hair has an important impact on the physiological growth of tea trees and the quality of tea, but the molecular mechanism of its formation is still unclear, which seriously hinders our in-depth understanding of the developmental biology of tea trees and the formation mechanism of tea quality

Method used

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  • A gene for regulating the formation of tea leaves and its application
  • A gene for regulating the formation of tea leaves and its application
  • A gene for regulating the formation of tea leaves and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1. Cloning and sequence structure analysis of CsMYB184 gene

[0018] Shucha, a national-level fine variety of tea tree, was planted in the Agricultural Industrial Park of Anhui Agricultural University, Hefei, Luyang District, Anhui Province, and the young leaves were used for RNA extraction. The total RNA was extracted with Trizol reagent (Invitrogen, USA) according to the instructions, and the RNA content and quality were detected with a spectrophotometer. Reverse transcription to generate the first strand: take 1 μg of RNA as a template, and synthesize cDNA according to the instructions of Promega M-MLV reverse transcriptase supporting reagent. After optimization, take an appropriate amount of reverse transcription product for subsequent PCR. The CsMYB184 gene was amplified by conventional PCR using cDNA as RT-PCR template. Upstream primer: (5'-ATGGCTCCGAAGAGCAGTGA-3'), downstream primer: (5'-TTACCATTTATCGGTAAGTGCC-3'). The 25 μL PCR reaction system was: 10...

Embodiment 2

[0023] Example 2. The correlation between the expression difference of CsMYB184 gene and the difference of tea millet distribution:

[0024] 1. The distribution and CsMYB184 gene expression in different tissues of tea plant

[0025] The national-level tea tree variety, Fuding Dabai, was planted in the Agricultural Industrial Park of Anhui Agricultural University, Hefei, Luyang District, Anhui Province. 13 tissues and organs were used to analyze the distribution and gene expression of tea leaves. The 13 tissues and organs include young shoots, one leaf, two leaves, three leaves, four leaves, old leaves, the first stem segment, the second stem segment, the third stem segment, the fourth stem segment, flowers, fruits, and roots. The distribution of tea hair of 13 varieties was photographed and recorded by stereo microscope or digital camera. These samples were also used for total RNA extraction and cDNA first-strand synthesis. The reverse transcription product (the first strand...

Embodiment 3

[0030] Example 3. Functional verification of CsMYB184 gene in Arabidopsis thaliana

[0031] 1. Construction of CsMYB184-pB2GW7 vector

[0032] Using the pMDTM 19-T Simple::CsMYB184 plasmid as a template, PCR amplification was performed with the following primers:

[0033] Upstream primer: (5'-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGGCTCCGAAGAGCAGTGA-3'),

[0034] Downstream primer: (5'-GGGGACCACTTTGTACAAGAAAGCTGGGTTTACCATTTATCGGTAAGTGCC-3'),

[0035] PCR products were recovered with 1% agarose gel electrophoresis bands. Using Gateway cloning technology, 1 μl of PCR recovery product was added, pDONR221 intermediate vector of equivalent quality was added, and finally 1 μL of BP Clonase Mix was added, and after overnight at room temperature, DH5α was transformed and sent for sequencing. The plasmids of the correct positive clones were sequenced, 1 μL of plasmid was taken and an equal amount of pB2GW7 overexpression vector was added. Finally, 1 μL of LR Clonase Mix was added. After o...

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Abstract

The invention discloses a gene for adjusting and controlling development of epidermal hair of camellia sinensis and quality formation of tea leaves and an application. The nucleotide sequence of a CsMYB184 gene is as shown in SEQID NO.1 in a sequence table, and the amino acid sequence of protein coded by the CsMYB184 gene is as shown in SEQID NO.2 in the sequence table. The expression pattern of the CsMYB184 is highly correlated to distribution of tea fine long hair; when the gene is used for overexpressing and transforming plants, the development defect phenotype of the epidermal hair of transgenic plants can completely return to normal; and through an antisense oligonucleotide technique, the expression of the CsMYB184 is restrained, the content of characteristic compounds in camellia sinensis leaves can be notably reduced. Cloning of the gene not only facilitates understanding a molecular mechanism for establishing the epidermal hair shape of the camellia sinensis, but also facilitates recognizing a molecular mechanism for forming important appearance and quality agronomic character namely tea fine long hair, a theory basis is provided for quality breeding of camellia sinensis, and the breeding process of specific camellia sinensis varieties is accelerated. The gene disclosed by the invention has great application value.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a tea tree MYB transcription factor gene and its encoded protein and application. Background technique [0002] Tea tree (Camellia sinensis (L.) O. Kuntze) is an important economic crop in my country. Tea is rich in amino acids, alkaloids, tea polysaccharides, polyphenols and volatile substances. The content and composition of these contents are different, which together endow tea with quality characteristics such as color, aroma and taste. Tea centimeter is an important agronomic trait of tea plants, and its rich and diverse secondary metabolites are not only the key to the formation of tea quality, but also play an important role in the response of tea plants to stress. Chahao is a single-cell type of epidermal hairs of tea, mainly distributed on the back of young bud leaves, the length varies according to the phyllotaxis, and falls off by itself as the young leave...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/20C12Q1/6895
CPCC07K14/415C12N15/8261C12Q1/6895C12Q2600/13C12Q2600/158
Inventor 赵剑李鹏辉徐玉婕张延瑞
Owner ANHUI AGRICULTURAL UNIVERSITY
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