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Molecular markers for simultaneously identifying restorability and authenticity of pepper male sterile three-line hybrid and identification method thereof

A male sterility and molecular marker technology, applied in the field of breeding, can solve the problems of short identification cycle, large number of markers, and high labor cost, and achieve the effect of reducing operation difficulty and identification cost.

Pending Publication Date: 2021-11-12
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the accuracy of field identification is high, the required identification cycle is long and the labor cost is high; while the identification at the biochemical level is likely to be unable to fully show the differences between varieties due to the low polymorphism of the protein; in contrast, using Molecular marker identification can directly identify the differences at the genetic level of varieties, and the identification cycle is short, which is the main development trend of future variety identification
[0005] Although there are many capsicum molecular markers that have been published, the results of molecular markers in practical applications are not ideal.
For example, although some molecular markers are closely related to the Rf map of the restorer gene, the identification results shown in practical applications are inconsistent with the phenotype, and even the target band may not be amplified.
There are also some molecular markers that are only developed for a single attribute of the authenticity or fertility of pepper varieties, and the single identification results of molecular markers cannot fully evaluate the usability of seeds
There are also molecular markers developed based on the whole genome of peppers. The number of markers provided is large, and there is no species specificity. In actual operation, multiple markers are required to be used together, and the workload of identification is heavy.
There are also some molecular markers that are only dominant markers, and cannot accurately distinguish between homozygous genotypes and heterozygous genotypes, and are easily interfered by the environment such as external temperature when applied.

Method used

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  • Molecular markers for simultaneously identifying restorability and authenticity of pepper male sterile three-line hybrid and identification method thereof
  • Molecular markers for simultaneously identifying restorability and authenticity of pepper male sterile three-line hybrid and identification method thereof
  • Molecular markers for simultaneously identifying restorability and authenticity of pepper male sterile three-line hybrid and identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Pepper prepare three-line hybrid male sterility and related materials 1. to be identified

[0040] Pepper male sterility three-line hybrid seeds ready to be identified 1.1

[0041] Pepper F1 random sample from the hybrid to be identified, for a total of not less than 100 filled seeds, were stored in refrigerator at 4 ℃.

[0042] Three male sterile line hybrid pepper 1.2 to be identified, the parent material preparation

[0043] 1.2.1 Seeds

[0044]Three-line male sterile hybrid pepper to be identified from the male, female in a random sample, from 20 to 40 grains were filled seeds, were stored in refrigerator at 4 ℃.

[0045] 1.2.2 blades

[0046] In the case of pepper seeds of the parent material is not available, the hybrid system may be farming, using the random number sampling method to extract plant material from the parent, coring blade 2-5 per sheet, self-sealing plastic to marker number bags, were stored at -20 ℃ refrigerator.

[0047] 2. The pepper seeds to be iden...

Embodiment 2

[0088] A rapid identification method for recovery and authenticity of pepper-male infertile three-strand hybrids simultaneously using molecular markers, including the following steps:

[0089] (1) Extract the pepper male infertile three-series hybrids and their parent, mother-in-the-art genomic DNA;

[0090] (2) Using molecular marking CR-S-01 or CR-S-02, PCR amplification, PCR product is performed, and the PCR product is subjected to polyacrylamide gel electrophoresis, observing the expansion strip;

[0091] (3) Compare the amplification strip of the hybrid seed with its parent, the maternal amplification strip, if a common dominant mark occurs in the hybrid, that is, in the hybrid seed, the mother is displayed and the parent The hierarchical strips are judged that the hybrids are the true hybrid of the parent and contain male infertility recovery genes (true toy and hybrid); if only the maternal amplification strip is displayed in the hybrid, it is judged that the hybrid The fak...

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Abstract

The invention discloses molecular markers for simultaneously identifying the restorability and authenticity of a pepper male sterile three-line hybrid and an identification method thereof. According to the invention, two molecular markers CR-S-01 and CR-S-02 capable of simultaneously identifying restorability and authenticity of the pepper male sterile three-line hybrid are designed by utilizing an SSR site in a pepper genome sequence, so that a simple, rapid and accurate method capable of simultaneously identifying authenticity, purity and restorability of the pigment processing type pepper male sterile three-line hybrid is established; and a basis can be provided for accelerating the improved variety inspection of the pigment processing type pepper male sterile three-line hybrid and the breeding of a male parent restorer. According to the invention, the two molecular markers are utilized to clearly distinguish true and false hybrids in the filial generation and meanwhile, identify whether the filial generation contains a male sterility restoring gene or not, so that the operation difficulty and the identification cost for identifying authenticity, purity and restorability of the hybrid can be greatly reduced.

Description

Technical field [0001] The present invention belongs to the technical field of breeding, particularly, to a hybrid simultaneous identification and recovery method of identifying molecular markers and three-line authenticity Male Sterility. Background technique [0002] Pepper, Capsicum genus Solanaceae, is an important crop and vegetable seasonings, because of its adaptability, unique flavor and rich nutritional value, high-value industries, widely grown in the country, is my country's traditional export products one. The type of chili pepper planting can be divided into fresh and processed Pepper, compared to the fresh pepper, chili and processing of high economic efficiency, long industrial chain, value-added products and great potential, such as the main source of pepper from the pepper coloring and processing red pigment, is widely used in processed foods as well as animal feed, cosmetics, and pharmaceuticals in. Although the prospects for industrial development and processin...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125Y02A50/30
Inventor 林多刘潇蔚杨延杰王辉朱文莹代佳汐
Owner QINGDAO AGRI UNIV