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CN-117448486-B - Application of SSR primer in evaluation of cauliflower resources

CN117448486BCN 117448486 BCN117448486 BCN 117448486BCN-117448486-B

Abstract

The invention belongs to the technical field of biology, and provides an application of an SSR primer in cauliflower germplasm resource identification, wherein the application comprises the application of the SSR primer in the identification of the variety of cauliflower, the genetic relationship evaluation, the cultivation of new germplasm of the cauliflower and the purity identification of the hybrid of the cauliflower. The 29 pairs of SSR primers screened by the invention have clear amplified band type, good stability and higher polymorphism, and are very suitable for variety identification, genetic relationship evaluation and purity identification of broccoli hybrid seeds for cultivating new broccoli germplasm.

Inventors

  • MENG SHUCHUN
  • XIA YANG
  • WANG HONGYANG
  • ZHANG CHENXI
  • YANG YESHENG
  • XU TAO
  • YU PINGBIN
  • LIU FAN
  • SONG SHUNHUA
  • HAN SHUO
  • XU XIULAN
  • SONG XIAOYU
  • WANG DEXIN
  • QIU YANHONG
  • ZHANG HAIJUN

Assignees

  • 北京市农林科学院

Dates

Publication Date
20260505
Application Date
20231127

Claims (8)

  1. 1. A kit for identifying cauliflower varieties, which contains 29 pairs of SSR primers numbered 1-58 in a sequence table; wherein the broccoli variety is any one of 84 broccoli varieties listed in table 1.
  2. 2. The kit of claim 1, wherein: The kit also contains at least one of PCR amplification buffer solution, dNTP, taq enzyme and DNA chromogenic dye.
  3. 3. A method of identifying a broccoli variety, the method comprising: SSR molecular marker detection is carried out on the cauliflower varieties with known variety information by 29 pairs of SSR primers numbered 1-58 in the sequence table, so that SSR fingerprint information of the cauliflower varieties with known variety information is obtained; performing SSR molecular marker detection on the cauliflower varieties to be detected by 29 pairs of SSR primers numbered 1-58 in the sequence table to obtain SSR fingerprint information of the cauliflower varieties to be detected; Comparing the SSR fingerprint information of the cauliflower variety to be detected with the SSR fingerprint information of the cauliflower variety with the known variety information, and judging that the cauliflower variety to be detected is consistent with the corresponding cauliflower variety with the known variety information if the similarity of the SSR fingerprint information of the cauliflower variety to be detected and the SSR fingerprint information of the cauliflower variety with the known variety information reaches 100%; wherein the broccoli variety is any one of 84 broccoli varieties listed in table 1.
  4. 4. A method according to claim 3, characterized in that: The SSR molecular marker detection comprises the following steps: the following PCR reaction system was used: 6. mu.L of DNA template, 2. Mu.L of 10XPCR buffer, 2. Mu.L of 10 Xdye, 0.4. Mu.L of 10 mM dNTPs, 0.3. Mu.L of 2.5 UTaq enzyme, 2. Mu.L of forward and reverse primer, and 9.3. Mu.L of ddH 2 O; And a PCR reaction procedure of 94℃for 5min, 94℃for 15S, 55℃for 15S, 72℃for 30S, 35 cycles, 72℃for 4 min, and 16℃for preservation.
  5. 5. A kit for evaluating the genetic relationship of cauliflowers, which comprises 29 pairs of SSR primers numbered 1-58 in a sequence table; wherein the broccoli variety is any one of 84 broccoli varieties listed in table 1.
  6. 6. The kit of claim 5, wherein: The kit also contains at least one of PCR amplification buffer solution, dNTP, taq enzyme and DNA chromogenic dye.
  7. 7. A method of broccoli affinity evaluation, the method comprising: performing SSR molecular marker detection on a plurality of broccoli varieties by 29 pairs of SSR primers numbered 1-58 in a sequence table to obtain SSR fingerprint information of the plurality of broccoli varieties; Calculating the genetic distance among the plurality of broccoli varieties according to the obtained SSR fingerprint information of the plurality of broccoli varieties; Judging the genetic relationship among a plurality of broccoli varieties according to the calculated genetic distance; wherein the broccoli variety is any one of 84 broccoli varieties listed in table 1.
  8. 8. A method for cultivating a novel germplasm of broccoli, the method comprising an operation of judging the relatedness between a plurality of broccoli varieties by adopting the method for evaluating the relatedness of broccoli according to claim 7, and then an operation of selecting two broccoli varieties farthest in relatedness as parent materials; wherein the broccoli variety is any one of 84 broccoli varieties listed in table 1.

Description

Application of SSR primer in evaluation of cauliflower resources Technical Field The invention belongs to the technical field of biology, and particularly relates to application of an SSR primer in cauliflower resource assessment. Background Broccoli (Brassica oleracea L. Var. Botrytis L.) is called cauliflower or cauliflower, belongs to Brassica of Brassicaceae, is developed from wild cabbage, and has delicious taste and rich nutritive value. Research shows that the cauliflower has the effects of preventing diseases, resisting cancer and the like, and is popular with people. The cauliflower industry has become one of the important vegetable industries in China, and is widely planted in Fujian, taiwan, guangdong, zhejiang and other provinces, and the number of main cultivated varieties is more than 170. The broccoli is originally produced in Europe, the broccoli variety resources in China are limited, genetic research is relatively backward, the selection of parents in the breeding process is dependent and blindness is caused, so that the new variety breeding of the broccoli is more difficult, the variety mixing problem is caused in the seed production process, and the phenomena of homonymous foreign matters and homonymous foreign matters occur in the market, so that the identification and genetic diversity analysis of the broccoli variety resources are developed, and the preservation and utilization of the broccoli variety resources are enhanced. Disclosure of Invention In order to solve the problems in the prior art, the invention provides an application of an SSR primer in cauliflower resource assessment. The first aspect of the invention provides a kit for identifying cauliflower varieties, which contains 29 pairs of SSR primers numbered 1-58 in a sequence table. The second aspect of the invention provides a method for identifying a broccoli variety, the method comprising: SSR molecular marker detection is carried out on the cauliflower varieties with known variety information by 29 pairs of SSR primers numbered 1-58 in the sequence table, so that SSR fingerprint information of the cauliflower varieties with known variety information is obtained; performing SSR molecular marker detection on the cauliflower varieties to be detected by 29 pairs of SSR primers numbered 1-58 in the sequence table to obtain SSR fingerprint information of the cauliflower varieties to be detected; Comparing the SSR fingerprint information of the cauliflower variety to be detected with the SSR fingerprint information of the cauliflower variety with the known variety information, and judging that the cauliflower variety to be detected is consistent with the corresponding cauliflower variety with the known variety information if the similarity of the SSR fingerprint information of the cauliflower variety to be detected and the SSR fingerprint information of the cauliflower variety with the known variety information is more than 98%. The third aspect of the invention provides a kit for evaluating the genetic relationship of cauliflowers, wherein the kit contains 29 pairs of SSR primers numbered 1-58 in a sequence table. In a fourth aspect, the present invention provides a method for evaluating the genetic relationship of cauliflowers, the method comprising: performing SSR molecular marker detection on a plurality of broccoli varieties by 29 pairs of SSR primers numbered 1-58 in a sequence table to obtain SSR fingerprint information of the plurality of broccoli varieties; Calculating the genetic distance among the plurality of broccoli varieties according to the obtained SSR fingerprint information of the plurality of broccoli varieties; And judging the genetic relationship among the plurality of broccoli varieties according to the calculated genetic distance. In a fifth aspect, the present invention provides a method for cultivating a novel germplasm of a broccoli, the method comprising an operation of judging the relatedness between a plurality of broccoli varieties by using the method for evaluating the relatedness of a broccoli provided in the fourth aspect of the present invention, and then selecting two broccoli varieties having the farthest relatedness as parent materials. The sixth aspect of the invention provides a kit for identifying the purity of the broccoli hybrid of the "Beijing-research holly 110", wherein the kit contains at least one SSR primer pair with the primer pair numbers of 1, 3, 15, 24, 25, 28 and 29 in a sequence table. The seventh aspect of the invention provides a kit for identifying the purity of the broccoli hybrid of the Ji-ping holly 100, wherein the kit contains at least one SSR primer pair with the primer pair numbers of 3, 6, 15 and 24 in a sequence table. The 29 pairs of SSR primers screened by the invention have clear amplified band type, good stability and higher polymorphism, and are very suitable for variety identification, genetic relationship evaluation and cultivation of novel broccoli ger