CN-119432839-B - Primer pair for detecting seed setting character of broccoli and related application of primer pair in breeding

Abstract

The invention discloses a primer pair for detecting the seed setting character of broccoli and related application thereof in breeding, relating to the field of plant breeding, wherein the primer pair comprises nucleotide sequences as shown in SEQ ID NO: 1-2, which can detect molecular markers closely linked with the seed setting character of broccoli and can be rapidly and effectively applied to the screening of broccoli breeding materials.

Inventors

• WU TING
• ZHAO HUI
• MA CUNFA
• YU YANG
• Zhong Liufeng
• XIAO JIANCHENG
• CHEN HUIZHONG
• YU YONGHUI

Assignees

• 浙江美之奥种业股份有限公司

Dates

Publication Date

20260508

Application Date

20241226

Claims (7)

1. 1. A primer pair is characterized by comprising an upstream primer and a downstream primer; The nucleotide sequence of the upstream primer comprises a sequence shown as SEQ ID NO. 1; The nucleotide sequence of the downstream primer comprises a sequence shown as SEQ ID NO. 2.
2. 2. A kit is characterized in that, comprising the primer pair of claim 1.
3. 3. The kit of claim 2, further comprising any one or more of PCR detection reagents, DNA polymerase and dNTPs.
4. 4. A method for detecting the setting character of broccoli is characterized by comprising the steps of carrying out PCR amplification on DNA of a sample to be detected by adopting the primer pair according to claim 1; The detection method further comprises the steps of judging a detection result according to a screening standard; the screening standard comprises the steps of judging that a sample is of a normal compact type if an amplification product is a single band, judging that the sample is of an unclamped type if the amplification product is a double band, wherein the single band is a single band of 200bp, and the double band is a double band of 200bp and 214 bp.
5. 5. The use of the primer pair according to claim 1 for the preparation of a product for detecting the set trait of broccoli.
6. 6. The use according to claim 5, wherein the product comprises a reagent combination or a kit.
7. 7. Use of the primer pair of claim 1 or the kit of claim 2 or 3 in the screening of broccoli breeding material or the breeding of broccoli.

Description

Primer pair for detecting seed setting character of broccoli and related application of primer pair in breeding Technical Field The invention relates to the field of plant breeding, in particular to a primer pair for detecting the setting character of broccoli and related application thereof in breeding. Background Broccoli (Brassica oleracea L.var.itica) is also known as broccoli, belongs to the brassica cabbage species of Brassicaceae, and has a nutritional value and a health care effect which are favored by consumers worldwide. At present, broccoli is mainly prepared by adopting a male sterile line, when female parent is fertility cytoplasm in the seed preparation process, self-bred seed setting or hybrid seed setting is normal, the expected seed quantity can be obtained, when female parent is transformed into sterile cytoplasm, seed yield is low due to abnormal seed collection or hybrid seed setting, so that the yield of broccoli varieties is difficult to meet market demands, and good new varieties are limited in popularization and application. The existing molecular regulation mechanism and gene positioning research aiming at the agronomic characters of broccoli is less. The research of the gene related to the fruit setting of the broccoli is basically blank except for the self-incompatible gene. In view of this, the present invention has been made. Disclosure of Invention The invention aims to provide a primer pair for detecting the setting character of broccoli and related application thereof in breeding. The invention is realized in the following way: In a first aspect, an embodiment of the invention provides a primer pair, which comprises an upstream primer and a downstream primer, wherein the nucleotide sequence of the upstream primer comprises a sequence shown as SEQ ID NO. 1 or is at least 80% identical to the sequence shown as SEQ ID NO. 1, and the nucleotide sequence of the downstream primer comprises a sequence shown as SEQ ID NO. 2 or is at least 80% identical to the sequence shown as SEQ ID NO. 2. In a second aspect, embodiments of the present invention provide a kit comprising the primer pair described in the previous embodiments. In a third aspect, the embodiment of the invention provides a method for detecting the setting property of broccoli, which comprises the step of carrying out PCR amplification on DNA of a sample to be detected by adopting the primer pair in the previous embodiment. In a fourth aspect, the present invention provides an application of the primer pair described in the previous embodiment or the kit described in the previous embodiment in preparing a product for detecting the fruit setting property of broccoli. In a fifth aspect, the present invention provides an application of the primer pair described in the previous embodiment or the kit described in the previous embodiment in screening broccoli breeding materials or breeding of broccoli. The invention has the following beneficial effects: The primer pair can detect the molecular marker closely linked with the seed setting character of the broccoli, and can be rapidly and effectively applied to screening of broccoli breeding materials. Drawings In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. FIG. 1 is a hybrid seed setting condition on a 7168 female parent (CMS) plant, wherein A is a plant setting diagram, B is a plant unset diagram, and C-D are seed pod setting and unset comparison diagrams; FIG. 2 is a breeding scheme for the broccoli combination 7168 as the female parent material; FIG. 3 is a BoseedF/BoseedR primer amplification 7168 female parent (CMS) N pool and Y Chi Duotai band pattern; FIG. 4 is a BoseedF/BoseedR primer amplification 7168 female (CMS) N-pool and Y-pool strip sequencing alignment; FIG. 5 is a plot of the running results of BoseedF/BoseedR primer amplification 7168 for 130 individuals of maternal (CMS) material; FIG. 6 is a gel running chart of BoseedF/BoseedR primer amplification stock shed 7168 female parent (fertility cytoplasm) for 152 individuals, wherein M is 100: 100bp DNALadder; FIG. 7 is a gel running chart of a total of 310 individual plants of BoseedF/BoseedR primer amplification stock shed 7168 female parent (CMS), wherein M is 100bp DNA Ladder. Detailed Description In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples a