CN-119842953-B - SSR molecular marker for resisting lesser leafhoppers of tea trees and application thereof

CN119842953BCN 119842953 BCN119842953 BCN 119842953BCN-119842953-B

Abstract

The invention relates to the technical field of molecular markers, and discloses an SSR molecular marker for tea tree leafhopper resistance and application thereof, wherein an SSR molecular marker primer amplifies a simple repeated sequence of a gene comprising one or more of CsTIFY1, csTIFY2, csTIFY3, csTIFY4, csTIFY5 and CsTIFY 6. According to the invention, the expression quantity of 6 CsTIFY genes is obviously up-regulated in the anti-lesser leafhoppers response process, further SSR locus retrieval is carried out on the 6 CsTIFY genes, 132 primers are designed, 5 SSR strips with obvious polymorphism and clear strips are screened through capillary electrophoresis, and the parting success rate of SSR molecular markers is found to be from 58.3 percent (CsTIFY 2-7) to 83.3 percent (CsTIFY-33) by combining the tea lesser leafhoppers resistance phenotype of 24 tea plant germplasm resources, so that a theoretical basis can be provided for early identification and molecular genetic breeding of tea tree lesser leafhoppers.

Inventors

LIU LIPING
LI DA
LI QINGSHENG
LU QINHUA
WANG YINGQI
Ren pi

Assignees

湖州市农业科学研究院(湖州市农业科技发展中心)

Dates

Publication Date: 20260508
Application Date: 20241224

Claims (2)

1. Application of SSR molecular marker primer of tea plant anti-lesser leafhopper in early identification of tea plant anti-lesser leafhopper and/or breeding of tea plant anti-lesser leafhopper, characterized in that the SSR molecular marker primer comprises one or more of CsTIFY-7 primer, csTIFY-11 primer, csTIFY3-6 primer, csTIFY-15 primer and CsTIFY-33 primer; the forward primer of the CsTIFY2-7 primer has a nucleotide sequence as shown in Seq ID No.1, and the reverse primer of the CsTIFY-7 primer has a nucleotide sequence as shown in Seq ID No. 2; The forward primer of the CsTIFY2-11 primer has a nucleotide sequence as shown in Seq ID No.3, and the reverse primer of the CsTIFY2-11 primer has a nucleotide sequence as shown in Seq ID No. 4; The forward primer of the CsTIFY-6 primer has a nucleotide sequence as shown in Seq ID No.5, and the reverse primer of the CsTIFY-6 primer has a nucleotide sequence as shown in Seq ID No. 6; The forward primer of the CsTIFY-15 primer has a nucleotide sequence as shown in Seq ID No.7, and the reverse primer of the CsTIFY-15 primer has a nucleotide sequence as shown in Seq ID No. 8; the forward primer of the CsTIFY4-33 primer has the nucleotide sequence shown as Seq ID No.9 and the reverse primer of the CsTIFY-33 primer has the nucleotide sequence shown as Seq ID No. 10.
2. The application of SSR molecular marker primers for resisting lesser leafhoppers of tea trees in the aspects of identification of plant germplasm resources of tea trees, research of genetic diversity, construction of genetic maps, relatedness of populations and evolution or genetic breeding is characterized in that the SSR molecular marker primers comprise one or more of CsTIFY-7 primers, csTIFY-11 primers, csTIFY3-6 primers, csTIFY3-15 primers and CsTIFY-33 primers; the forward primer of the CsTIFY2-7 primer has a nucleotide sequence as shown in Seq ID No.1, and the reverse primer of the CsTIFY-7 primer has a nucleotide sequence as shown in Seq ID No. 2; The forward primer of the CsTIFY2-11 primer has a nucleotide sequence as shown in Seq ID No.3, and the reverse primer of the CsTIFY2-11 primer has a nucleotide sequence as shown in Seq ID No. 4; The forward primer of the CsTIFY-6 primer has a nucleotide sequence as shown in Seq ID No.5, and the reverse primer of the CsTIFY-6 primer has a nucleotide sequence as shown in Seq ID No. 6; The forward primer of the CsTIFY-15 primer has a nucleotide sequence as shown in Seq ID No.7, and the reverse primer of the CsTIFY-15 primer has a nucleotide sequence as shown in Seq ID No. 8; the forward primer of the CsTIFY4-33 primer has the nucleotide sequence shown as Seq ID No.9 and the reverse primer of the CsTIFY-33 primer has the nucleotide sequence shown as Seq ID No. 10.

Description

SSR molecular marker for resisting lesser leafhoppers of tea trees and application thereof Technical Field The invention relates to the technical field of molecular markers, in particular to an SSR molecular marker for tea tree anti-leafhoppers and application thereof. Background Tea tree (CAMELLIA SINENSIS) is a worldwide important commercial crop that faces a threat from a variety of pests, with tea leafhoppers being of great concern due to their high degree of attack on tea tree shoots and young leaves. The rapid propagation and strong selective invasiveness of the tea leafhoppers seriously threaten the growth and yield of the tea. The TIFY transcription factor has a wide and important function in plant growth and development. Studies have shown that TIFY family members affect stress resistance and hormone response mechanisms in plants by modulating the Jasmonic Acid (JA) signaling pathway. The TIFY family can be divided into 4 subfamilies according to the type of domain it contains, TIFY, ZML (ZIM & ZIM-like), JAZ (jasmonate-ZIM-domain) and PPD (PEAPOD). In recent years, the role of the TIFY transcription factor family in plant pest control mechanisms has attracted considerable attention. JAZ proteins of TIFY family are key inhibitors in the JA signaling pathway, which can modulate the JA signaling pathway to increase the insect-resistant response of plants. In alfalfa (Medicago sativa l.), the expression of MsTIFY in anti-thrips and easy alfalfa varieties was significantly up-regulated 14 days after thrips infestation. A simple repeat sequence (SSR) is taken as a repeat sequence in a genome, and has the characteristics of 1-6 nucleotide units, such as (CA) n, (AT) n, (GGC) n and the like. Because of the advantages of abundant quantity, good polymorphism, low use cost, simple operation and the like, SSR molecular markers are widely applied to the fields of plant genetic map construction, genetic diversity analysis, gene positioning, cloning and the like. Disclosure of Invention In order to solve the existing problems, the invention discloses an SSR molecular marker for resisting leafhoppers of tea trees and application thereof. According to the invention, the expression quantity of 6 CsTIFY genes is obviously up-regulated in the anti-lesser leafhoppers response process, further SSR locus retrieval is carried out on the 6 CsTIFY genes, 132 primers are designed, 5 SSR strips with obvious polymorphism and clear strips are screened through capillary electrophoresis, and the parting success rate of SSR molecular markers is found to be from 58.3 percent (CsTIFY 2-7) to 83.3 percent (CsTIFY-33) by combining the tea lesser leafhoppers resistance phenotype of 24 tea plant germplasm resources, so that a theoretical basis can be provided for early identification and molecular genetic breeding of tea tree lesser leafhoppers. In one aspect, the invention provides an SSR molecular marker primer for tea tree anti-leafhoppers, which amplifies a simple repeated sequence of a gene comprising one or more of CsTIFY, csTIFY, csTIFY3, csTIFY4, csTIFY5 and CsTIFY 6. The CsTIFY, csTIFY2, csTIFY, csTIFY4, csTIFY5 and CsTIFY6 belong to TIFY family member genes, wherein CsTIFY1 has the gene number GWHGACFB 004081), csTIFY2 has the gene number GWHGACFB028923, csTIFY3 has the gene number GWHGACFB013026, csTIFY4 has the gene number GWHGACFB021202, csTIFY5 has the gene number GWHGACFB022136, csTIFY6 has the gene number GWHGACFB005241, and the genes can be inquired from a tea plant genome database http:// tpia teaplans. The invention re-annotates on-line published tea plant anti-lesser leafhopper transcriptome data (PRJNA 553681) to 'Longjing 43' genome, adopts DESeq2 software package to carry out differential expression gene screening analysis (q value <0.005&|log2 (Fold change) > 1|) to obtain 3795 Differential Expression Genes (DEGs), carries out KEGG pathway enrichment analysis to screen out anti-lesser leafhopper key genes, and finds out that 6 tea plant TIFY family member genes CsTIFY1, csTIFY2, csTIFY3, csTIFY4, csTIFY5 and CsTIFY are all obviously up-regulated, which indicates that CsTIFYs genes have the same expression variation trend in the anti-lesser leafhopper response process, and speculates that CsTIFY family members regulate the anti-lesser leafhopper mechanism of tea plants through JA signal transmission. The subsequent test carries out SSR locus retrieval and primer design aiming at CsTIFY family members, and verifies that the primer for amplifying CsTIFY family member genes can be used as an SSR molecular marker primer for resisting the leafhoppers of tea trees, and can be used for typing whether the leafhoppers of tea trees are resisting. Therefore, based on the effect of the 6 tea tree TIFY family member genes found by the invention in the tea tree anti-lesser leafhopper pathway, the SSR molecular marker primers can be designed for the 6 tea tree TIFY family member genes, while the SSR sites in the 6 tea tree TIFY