CN-122012797-A - KASP molecular marker related to soybean single plant pod number and grain number characters and application thereof

CN122012797ACN 122012797 ACN122012797 ACN 122012797ACN-122012797-A

Abstract

The invention discloses a KASP molecular marker related to soybean single plant pod number and grain number characters and application thereof, belonging to the technical fields of soybean agronomic character molecular marker development and molecular marker assisted breeding. The invention discloses a KASP molecular marker related to soybean single plant pod number and grain number character, which is positioned at 45649932 bp site of soybean genome Wm82.a4.v1 version 19 chromosome, and the genotype is A or T. The invention provides a novel and simple molecular marker and auxiliary selection method, which is suitable for rapid screening of the number of soybean single plants and the relative height of the number of grains and auxiliary breeding of the molecular marker. The molecular marker can detect genotypes of different single-plant pod numbers and grain number materials in natural populations, can also be used for molecular auxiliary selection breeding of different single-plant pod numbers and grain number soybean materials, replaces large-scale phenotype screening, reduces field workload, and remarkably improves screening accuracy and efficiency.

Inventors

LIU XIULIN
ZHANG SHIYAO
XIA XIAOYU
YUAN RONGQIANG
REN HONGLEI
ZHANG CHUNLEI
ZHANG BIXIAN
WANG XUEYANG
ZHANG FENGYI
ZHAO KEZHEN

Assignees

黑龙江省农业科学院大豆研究所(黑龙江农业科技杂志社)

Dates

Publication Date: 20260512
Application Date: 20260403

Claims (6)

1. A KASP molecular marker associated with soybean pod number and grain number traits characterized in that the KASP molecular marker genotype is a or T located at 45649932 bp of chromosome 19 of soybean genome wm82.a4.v1.
2. The KASP molecular marker related to soybean pod number and grain number characteristics according to claim 1, wherein when the KASP molecular marker genotype is A, the sequence of 50bp before and after is shown as SEQ ID NO.1, the corresponding phenotype is low pod number and grain number type, and when the KASP molecular marker genotype is T, the sequence of 50bp before and after is shown as SEQ ID NO.2, the corresponding phenotype is high pod number and grain number type.
3. The KASP molecular marker primer combination for identifying the pod number and grain number characters of soybean is characterized by comprising the following primer sequences: forward primer F-FAM: 5’-GAAGGTGACCAAGTTCATGCTCCTGTGTGCGTAGCCTCACAT-3’;SEQ ID NO.3; forward primer F-HEX: 5’-GAAGGTCGGAGTCAACGGATTCCTGTGTGCGTAGCCTCACAA-3’;SEQ ID NO.4; Reverse primer R: 5’-GTCTCTCTTCTGTGTCTGTTATTAGTGG-3’;SEQ ID NO.5。
4. a kit for identifying soybean pod number and grain number traits comprising the KASP molecular marker primer combination of claim 3.
5. Use of a KASP molecular marker according to any one of claims 1-2 or a KASP molecular marker primer combination according to claim 3 or a kit according to claim 4 for the identification or co-identification of soybean pod number and grain number traits.
6. A method for identifying the pod number and grain number characteristics of a soybean plant, comprising the following steps: (1) Extracting genomic DNA of a soybean sample to be detected, carrying out PCR amplification on the genomic DNA of the soybean sample on a ABI Stepone PCR instrument by adopting the KASP molecular marker primer combination of claim 3, detecting fluorescent signals by utilizing a ABI Stepone PCR instrument, and analyzing genotyping; (2) Judging according to genotype results, wherein when the genotyping result of the sample is consistent with the low-single-plant pod number and grain number soybean material genotype AA, the identified soybean sample is expressed as the low-single-plant pod number and grain number character, and when the genotyping result of the sample is consistent with the high-single-plant pod number and grain number soybean material genotype TT, the identified soybean sample is expressed as the high-single-plant pod number and grain number character.

Description

KASP molecular marker related to soybean single plant pod number and grain number characters and application thereof Technical Field The invention relates to the technical field of soybean agronomic trait molecular marker development and molecular marker assisted breeding, in particular to a KASP molecular marker related to soybean single plant pod number and grain number traits and application thereof. Background Soybeans are taken as important economic grain and oil crops in China, and are favored in the global scope by virtue of rich nutritional values and wide medicinal values. It contains basic nutrients of isoflavone, high-quality protein, several essential amino acids, unsaturated fatty acid, etc. and is rich in various bioactive matters, such as soybean isoflavone, soybean polypeptide, soybean phospholipid, soybean oligosaccharide, soybean saponin, etc. Modern medical research proves that the active ingredients have remarkable effects of resisting tumor, reducing blood fat, preventing cardiovascular diseases and the like, and have great development potential in the fields of functional foods and medicines. The soybean yield traits are genetically represented as complex traits commonly controlled by a plurality of Quantitative Trait Loci (QTL), and the traditional breeding method has the inherent defects of long period, low efficiency, limited yield increase amplitude and the like. With the breakthrough development of molecular biology technology, in particular to the increasingly mature high-throughput sequencing technology and gene editing technology, the gene fine regulation technology provides a brand new way for realizing the precise polymerization and the efficient utilization of excellent alleles. The technological breakthrough is regarded as a core key technology for cultivating breakthrough soybean varieties, and is also a necessary way for improving soybean breeding capacity in the future. To achieve this objective, first, the work of mining and accumulating excellent alleles needs to be systematically developed, while high-precision molecular markers are developed in a matched manner—these basic works constitute prerequisites for fine regulation of genes and multi-gene polymerization. Specifically, the complete genotype-phenotype association database is established by using the technical means of whole genome association analysis (GWAS), transcriptome and the like, a high-density SNP marker system covering the whole genome is developed, and a novel breeding system combining molecular Marker Assisted Selection (MAS) and Genome Selection (GS) is constructed. Through the systematic work, the mode transition from 'empirical breeding' to 'precise design breeding' is finally realized, and a powerful technological support is provided for the high-quality development of soybean industry in China. The number of single plants and the number of grains of soybean are taken as direct component characters for forming the yield, and the genetic analysis of the soybean has important significance for high-yield breeding. Research shows that there is a significant positive correlation between the pod number and grain number of soybean plants, and both show higher genetic transmission. It is documented in the Soybase database that more than 150 pod number related QTLs and more than 120 pod number related QTLs have been located, and these sites are widely distributed on all soybean chromosomes except chromosome 12, with densely distributed regions of multiple pod number QTLs on chromosomes 6,8, 13, 17 and 19. Zhang Lei et al (2018) identified 7 SNP sites in the cultivated soybean population that were significantly associated with the number of pods per plant by whole genome association analysis, located on chromosome 5, 9 and 16, respectively, and found that two of these sites were adjacent to known flowering genes, revealing the regulatory effect of the photoperiod pathway on pod number formation. Zhang Qingqi et al (2020) further utilized chromosome fragment substitution line population, finely positioned to a main effect QTL for controlling the number of single plant pods and the number of grains simultaneously in the 17 th chromosome qPod17.1 interval, and the synergistic allele of the main effect QTL can increase the number of single plant pods by 12.3%, increase the number of grains by 9.7%, so that an accurate target point is provided for molecular marker assisted selection. The total yield of soybean in 2021 is 1640 ten thousand tons, but the continuous improvement of the unit yield of soybean is still a core target of breeding work in face of huge population demand and economic development demand. The molecular Marker Assisted Selection (MAS) and Genome Selection (GS) technology can accurately position the number of single plants and the number of grains QTL, so that the breeding efficiency can be remarkably improved, and the cultivation process of high-yield soybean varieties can be accelerated. Mea