CN-122012790-A - KASP molecular marker primer group and kit for detecting genetic locus related to environmental adaptation key characters of tea trees and application of kit

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 environmental adaptation key traits of tea trees, a kit and application thereof. The invention provides 2 InDel sites 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 InDel sites relate to frost resistance (Frost Damage Index, FDI) 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 environment adaptation of the tea tree 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

20260512

Application Date

20251205

Claims (5)

1. 1. A KASP molecular marker primer set for detecting genetic loci associated with environmental adaptation key traits of tea trees, wherein the environmental adaptation key traits of tea trees are strong freezing resistance characteristics of tea trees and high photosynthetic efficiency characteristics of tea trees, the primer set comprising: InDel2 primer pair with SEQ ID NO of 10-12, and InDel3 primer pair is 16-18 SEQ ID NO.
2. 2. A KASP molecular marker primer set for detecting a genetic locus associated with an environmental adaptation key trait of tea tree 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. 3. A kit comprising a set of KASP molecular marker primers for detecting a genetic locus associated with an environmental adaptation key trait of camellia sinensis as defined in claim 1.
4. 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 environmentally adapted key trait characteristics, in particular: a) Screening tea tree germplasm with strong freezing resistance, or/and B) Screening tea tree germplasm with high photosynthetic efficiency.
5. 5. A method for detecting environmental adaptation key trait genotyping of tea tree 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, performing identification analysis according to FAM and HEX fluorescent signal results after PCR amplification, and determining the genotyping of the environmental adaptation key characters of the tea tree to be detected, wherein the environmental adaptation key characters of the tea tree are the strong freezing resistance characteristic and the high photosynthetic efficiency characteristic of the tea tree.

Description

KASP molecular marker primer group and kit for detecting genetic locus related to environmental adaptation key characters of tea trees 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 environmental adaptation key traits of tea trees, a kit and application thereof. 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 tea trees and can be applied to rapid screening of high-resistance good varieties 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 environmental adaptation key traits of tea trees, wherein the environmental adaptation key traits of tea trees are strong freezing resistance characteristics and high photosynthetic efficiency characteristics of tea trees, and the primer group comprises: InDel2 primer pair with SEQ ID NO of 10-12, and InDel3 primer pair is 16-18 SEQ ID NO. 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 invention provides a kit containing the KASP molecular mar