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CN-122012783-A - KASP molecular marker and primer of balsam pear and application of KASP molecular marker and primer

CN122012783ACN 122012783 ACN122012783 ACN 122012783ACN-122012783-A

Abstract

The invention relates to a KASP molecular marker and a primer of balsam pear and application thereof, belonging to the technical field of molecular biology. The invention provides a KASP molecular marker of balsam pear, which comprises one of MC07-0177 molecular marker and MC07-1544 molecular marker. According to the invention, two KASP molecular markers closely related to the bitter gourd fusarium wilt resistance genes are obtained through screening, experiments show that the two KASP molecular markers can distinguish the bitter gourd fusarium wilt resistance single plants from the fusarium wilt susceptible single plants, the consistency of detection results and actual investigation results is more than 90%, and the KASP molecular markers are used for screening the bitter gourd fusarium wilt resistance, so that the method is simple, convenient, rapid and efficient, and lays a foundation for bitter gourd fusarium wilt resistance molecular breeding.

Inventors

  • JU QIANQIAN
  • FENG CHENGCHENG
  • HUANG RUKUI
  • HUANG XIONGJUAN
  • CHEN XIAOFENG
  • HUANG YUHUI
  • LIANG JIAZUO
  • LIU XINGLIAN

Assignees

  • 广西壮族自治区农业科学院

Dates

Publication Date
20260512
Application Date
20260306

Claims (10)

  1. 1. A KASP molecular marker of balsam pear, characterized in that the KASP molecular marker comprises one of MC07-0177 molecular marker and MC07-1544 molecular marker; The MC07-0177 molecular marker is positioned at 13770177 th chromosome of Momordica Charantia No. 7, and the mutation type is A/G; The MC07-1544 molecular marker is positioned at 14121544 th chromosome of Momordica Charantia No. 7, and the mutation type is C/T.
  2. 2. The KASP molecular marker according to claim 1, wherein when the genotype at the MC07-0177 molecular marker is AA, the balsam pear plant is a homozygous wilt-resistant plant, when the genotype is GG, the balsam pear is a homozygous wilt-resistant plant, and when the genotype is GA, the balsam pear is a heterozygous plant; And when the genotype of the MC07-1544 molecular marker is CC, the balsam pear plant is a homozygous anti-wilt plant, when the genotype is TT, the balsam pear plant is a homozygous anti-wilt plant, and when the genotype is CT, the balsam pear plant is a heterozygous plant.
  3. 3. A primer set for detecting the KASP molecular marker according to claim 1, wherein the primer set comprises a primer set for detecting the MC07-0177 molecular marker and a primer set for detecting the MC07-1544 molecular marker.
  4. 4. The primer group according to claim 3, wherein the primer group for detecting MC07-0177 molecular marker comprises a forward primer 1, a forward primer 2 and a universal reverse primer 1; the nucleotide sequence of the forward primer 1 is shown as SEQ ID NO. 1; The nucleotide sequence of the forward primer 2 is shown as SEQ ID NO. 2; the nucleotide sequence of the universal reverse primer 1 is shown as SEQ ID NO. 3.
  5. 5. The primer group of claim 4, wherein the 5' end of the forward primer 1 is connected with a FAM fluorescent tag sequence, and the nucleotide sequence of the FAM fluorescent tag sequence is shown as SEQ ID NO. 7; and the 5' end of the forward primer 2 is connected with a HEX fluorescent tag sequence, and the HEX fluorescent tag sequence is shown as SEQ ID NO. 8.
  6. 6. The primer set of claim 3, wherein the primer set for detecting the MC07-1544 molecular marker comprises a forward primer 3, a forward primer 4 and a universal reverse primer 2; The nucleotide sequence of the forward primer 3 is shown as SEQ ID NO. 4; the nucleotide sequence of the forward primer 4 is shown as SEQ ID NO. 5; the nucleotide sequence of the universal reverse primer 2 is shown as SEQ ID NO. 6.
  7. 7. The primer group of claim 6, wherein the 5' end of the forward primer 3 is connected with a FAM fluorescent tag sequence, and the nucleotide sequence of the FAM fluorescent tag sequence is shown as SEQ ID NO. 7; And the 5' end of the forward primer 4 is connected with a HEX fluorescent tag sequence, and the HEX fluorescent tag sequence is shown as SEQ ID NO. 8.
  8. 8. Use of a KASP molecular marker according to claim 1 or 2 or a primer set according to any one of claims 3 to 7 in breeding of balsam pear anti-wilt varieties.
  9. 9. Use of a KASP molecular marker according to claim 1 or 2 or a primer set according to any one of claims 3 to 7 for the preparation of a product for screening homozygous plants of balsam pear against wilt.
  10. 10. A method for detecting a homozygous plant of momordica charantia against wilt, the method comprising the steps of: Extracting DNA of the balsam pear to be tested, using the DNA as a template, adopting a primer group to amplify to obtain an amplified product, distinguishing the genotype of the amplified product, and detecting the homozygous plant of the balsam pear with resistance to the fusarium wilt according to the genotype; the primer set is the primer set according to any one of claims 3 to 7.

Description

KASP molecular marker and primer of balsam pear and application of KASP molecular marker and primer Technical Field The invention relates to the technical field of molecular biology, in particular to a KASP molecular marker, a primer and application of the KASP molecular marker. Background Balsam pear (Momordica charantia L.) is an important health-care vegetable of Cucurbitaceae, and its pulp is rich in vitamin C, carotenoid and mineral substances, and chemical components such as saponin, and has anticancer, blood sugar lowering, antiinflammatory, antibacterial and immunity enhancing effects. In recent years, due to the fact that people have a deep understanding of the medicinal and health-care effects of balsam pears, the consumer population of balsam pears is gradually expanded to the whole Asia and other parts of the world, and the balsam pears become global vegetables. The planting area of balsam pear in Guangxi every year reaches more than 60 ten thousand mu, and the serious occurrence of fusarium wilt is caused by the intensive planting in summer at high temperature and high humidity and in various places, and the incidence rate of serious disease areas reaches 85 percent. Wilt is one of the most destructive diseases in the major areas of balsam pear. Fusarium wilt (Fusarium wilt) is an important destructive soil-borne disease caused by the specialization of fusarium oxysporum balsam pear. After the balsam pear is infected with fusarium wilt, the leaves gradually become yellow and fusarium wilt from bottom to top due to the blockage of the catheter and the action of toxin until the whole plant dies. According to the investigation and research of the former, the average incidence rate of the fusarium wilt of the bitter gourd in the continuous cropping area of the facility reaches 13% -51% and 40% -80% respectively in the full bloom period and the full fruit period, so that serious yield reduction and even harvest failure can be caused, and huge economic loss is brought to the production of the bitter gourd. The pathogenic bacteria of the fusarium wilt of the bitter melon have strong viability in soil and host specialization, are difficult to control, and are the most effective and environment-friendly measures for breeding disease-resistant varieties. The comprehensive prevention and control of the fusarium wilt of the bitter melon still faces the technical bottleneck at present that the conventional chemical prevention and control is difficult to realize effective control after disease outbreak, and the grafting technology has application limitations of time and labor consumption, high economic cost and the like although having prevention and control effects. In contrast, breeding of disease-resistant varieties is still considered as the most economic-benefit prevention and control strategy. With the rapid development of molecular biology technology, molecular breeding technology greatly accelerates the progress of disease-resistant breeding and obviously shortens the period of breeding disease-resistant varieties. The DNA molecular marker technology plays an extremely important role in crop disease-resistant genetic breeding due to the characteristics of high polymorphism, convenient detection, rapidness, accuracy and the like. At present, researches on the fine positioning of major QTL of cucumber fusarium wilt and related molecular markers thereof are not perfect. Based on this, the present invention has been proposed. Disclosure of Invention The invention aims to provide a KASP molecular marker, a primer and application thereof for balsam pear, which are used for solving the problem of imperfect research on molecular markers related to fusarium wilt in balsam pear in the prior art. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides a KASP molecular marker of balsam pear, which comprises one of MC07-0177 molecular marker and MC07-1544 molecular marker; The MC07-0177 molecular marker is positioned at 13770177 th chromosome of Momordica Charantia No. 7, and the mutation type is A/G; The MC07-1544 molecular marker is positioned at 14121544 th chromosome of Momordica Charantia No. 7, and the mutation type is C/T. Preferably, when the genotype of the MC07-0177 molecular marker is AA, the balsam pear plant is a homozygous wilt-resistant plant, when the genotype is GG, the balsam pear is a homozygous wilt-resistant plant, and when the genotype is GA, the balsam pear is a heterozygous plant; And when the genotype of the MC07-1544 molecular marker is CC, the balsam pear plant is a homozygous anti-wilt plant, when the genotype is TT, the balsam pear plant is a homozygous anti-wilt plant, and when the genotype is CT, the balsam pear plant is a heterozygous plant. The invention provides a primer group for detecting KASP molecular markers, which comprises a primer group for detecting MC07-0177 molecular markers and a primer group for d