CN-121992139-A - KASP molecular marker primer group and kit for detecting genetic locus related to key character of secondary metabolite of tea tree and application of kit

CN121992139ACN 121992139 ACN121992139 ACN 121992139ACN-121992139-A

Abstract

The invention belongs to the field of molecular markers, and particularly relates to a KASP molecular marker primer group for detecting genetic loci related to key traits of secondary metabolites of tea trees, a kit and application of the kit. The invention provides 2 SNP loci which are identified based on whole genome association analysis (GWAS) and are obviously related to key agronomic traits and characteristic flavors of tea trees, wherein the loci relate to the content characteristics of secondary metabolites of the tea trees. Based on the sites, the invention develops a corresponding KASP primer group for detecting the genotyping of the key characters of the secondary metabolites of the tea trees and assisting in selective breeding. The system has the advantages of high throughput, high accuracy, low cost and the like, not only enriches molecular marker resources of tea trees, but also can be used for early selection and excellent single plant screening in molecular breeding of the tea trees, and remarkably improves breeding efficiency.

Inventors

HAO XINYUAN
TANG JUNWEI
HUANG CHAO
WANG XINCHAO

Assignees

中国农业科学院茶叶研究所

Dates

Publication Date: 20260508
Application Date: 20251205

Claims (5)

1. A primer set for detecting a KASP molecular marker for a genetic locus associated with a key trait of a secondary metabolite of camellia sinensis, wherein the key trait of environmental adaptation of camellia sinensis is a high methylated catechin content trait of camellia sinensis and a high theanine glucoside content trait of camellia sinensis, the primer set comprising: SNP4 primer pair of 19-21 SEQ ID NO, and SNP5 primer pair, SEQ ID NO. 22-24.
2. A KASP molecular marker primer set for detecting a genetic locus associated with a key trait of tea tree secondary metabolites as claimed in claim 1 wherein two allele-specific forward primers of said KASP molecular marker primer set are respectively linked to FAM fluorescent markers or HEX fluorescent markers.
3. A kit comprising a set of KASP molecular marker primers for detecting a genetic locus associated with a key trait of tea tree secondary metabolites as defined in claim 1.
4. Use of a KASP molecular marker primer set according to claim 1 or a kit according to claim 3 for screening tea tree germplasm with secondary metabolite key trait characteristics, in particular: a) Screening tea plant germplasm with high methylated catechin content, and/or B) Tea tree germplasm with high theanine glucoside content is screened.
5. A method for detecting genotyping of a key trait of a secondary metabolite of camellia sinensis using the KASP molecular marker primer set of claim 1 or the kit of claim 3, comprising the steps of: s.1, extracting genome DNA of tea tree germplasm resources to be detected; s.2, carrying out PCR amplification on the genomic DNA of the tea tree germplasm resource to be detected by using the KASP molecular marker primer group of claim 1 or the kit of claim 3 in a KSAP genotyping instrument; And S.3, identifying and analyzing according to FAM and HEX fluorescent signal results after PCR amplification, and determining the genotyping of the key characters of the secondary metabolites of the tea tree to be detected, wherein the key characters of the secondary metabolites of the tea tree are the high-methylation catechin content characteristic and the high-theanine glucoside content characteristic of the tea tree.

Description

KASP molecular marker primer group and kit for detecting genetic locus related to key character of secondary metabolite of tea tree and application of kit The application relates to a division application, the original application number is CN2025118212953, the application date is 2025, 12 and 05 days, and the application name is a KASP molecular marker primer group for detecting the related genetic locus of key characters of tea trees, a kit and application thereof. Technical Field The invention belongs to the field of molecular markers, and particularly relates to a KASP molecular marker primer group for detecting genetic loci related to key traits of secondary metabolites of tea trees, a kit and application of the kit. Background Tea tree (CAMELLIA SINENSIS) is an important commercial crop whose production is a highly global agricultural industry with annual average yields of over 2000 billion dollars and over 663 tons. Tea is not only widely consumed in the world, but also has remarkable health care functions including antioxidation, anti-inflammatory, lipid-lowering, chronic disease prevention and the like due to various bioactive components (such as tea polyphenol, theanine, caffeine and the like) contained in the tea. With the increase of global consumer health consciousness and the diversification of market demands, the demands for tea quality and special flavor are increasingly raised, and higher demands are put on improvement of tea tree varieties. The key agronomic characters of tea trees, such as high yield and stable yield, disease resistance and stress resistance (including cold resistance, drought resistance, disease and insect resistance and the like) and environmental adaptability, are directly related to the yield stability of tea and the economic benefit of planting. Meanwhile, secondary metabolites of tea, such as catechin (particularly epigallocatechin gallate (EGCG)), theanine, caffeine, aroma substances, etc., are core factors for determining tea quality, flavor characteristics and health functions. Most of these traits belong to quantitative traits, are controlled by multiple genes, and are susceptible to environmental factors (such as climate, soil, cultivation measures, etc.), exhibiting complex genetic mechanisms. Traditional tea tree breeding mainly depends on phenotype selection, and individuals with excellent properties are screened through methods of hybridization, mutation breeding, systematic breeding and the like in combination with long-term field observation and evaluation. However, this approach has significant limitations such as lengthy periods, inefficiency, low accuracy in phenotypic selection, and ambiguous targets. These factors severely restrict the progress of tea tree breeding work and the breeding efficiency of new varieties. In recent years, with the accumulation of genomic data of tea trees, molecular marker-based assisted selective breeding has become an effective means for improving breeding efficiency. However, the number of markers commonly used in the prior art, such as SSR, RFLP and the like, is limited, the flux is low, the cost is high, and the relevance of most markers and traits is not verified by multiple environments, so that the large-scale application of the markers is limited. The KASP (Kompetitive ALLELE SPECIFIC PCR) technique is a PCR-based SNP typing technique that enables high throughput, low cost and automated large scale sample typing by allele-specific primer and fluorescent signal detection. The development of a molecular marker system based on the KASP technology is helpful for breaking through the bottleneck of tea tree breeding and pushing the tea tree breeding to develop accurately and efficiently. Disclosure of Invention Aiming at the problems of limited number of molecular markers of tea trees, low detection efficiency, poor universality and the like in the prior art, the invention aims to provide a set of molecular marker system developed based on GWAS identification and KASP technology, which can be used for rapid and accurate typing of key characters of secondary metabolites of tea trees, and can be applied to rapid screening of excellent varieties of high-content substances in molecular breeding of tea trees. The invention is realized by the following technical scheme: The first aspect of the invention provides a KASP molecular marker primer group for detecting genetic loci related to key traits of secondary metabolites of tea trees, wherein the key traits of environmental adaptation of the tea trees are high-methylation catechin content characteristics and high-theanine glucoside content characteristics of the tea trees, and the primer group comprises: SNP4 primer pair of 19-21 SEQ ID NO, and SNP5 primer pair, SEQ ID NO. 22-24. Further, FAM fluorescent markers or HEX fluorescent markers are respectively connected to two allele-specific forward primers in the KASP molecular marker primer group. In a second aspect, the inventi