PCR primer set for identification or screening of cabbage hybrid lethal parent type 2 and its application
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A primer set and a technology for cabbage, applied in the fields of molecular biology and plant genetics and breeding, can solve problems such as production loss of cabbage, and achieve the effects of improving breeding efficiency, good stability and high stability
Active Publication Date: 2022-04-15
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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In recent years, there have been more and more studies on the lethality of plant hybrids, but in cruciferous crops, most of them are caused by intergeneric hybrids caused by distant hybridization, and the cases of intraspecific hybrid lethality are only in Arabidopsis thaliana. and Kale have been studied on
Cabbage hybrid lethality is an extreme abnormal phenomenon in the growth and development of cabbage, which will bring losses to the production of cabbage
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Embodiment 1
[0041] The inventors compared and analyzed the resequencing results of the cabbage hybrid lethal parent type 1 inbred line 09-211 and the cabbage hybrid lethal parent type 2 inbred line 09-222, and detected the cabbage hybrid lethal parent type 2 inbred line 09-222 There is a SNP variation at positions 46, 333, and 244 on chromosome C04, which was confirmed by first-generation sequencing. C04: position 46,333,244 represents the physical position 46,333,244 on the chromosome of Brassica oleracea C04, which is determined based on the whole genome sequence of Brassica oleracea "TO1000", which is recorded in the Ensembl Plants database (URL http: / / plants.ensembl.org / index.html).
[0042] In order to verify the correlation between this variation and the hybrid lethal parent type 2, intercept its upstream and downstream 50bp sequence (SEQ ID NO.1), according to the KASP design principle, design KASP primers on the Ploy Marker website, and design based on the variation site to get ...
Embodiment 2
[0047] Example 2. Correlation verification between PCR markers developed by the present invention and hybrid lethal parent type 2
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Abstract
The invention belongs to the technical field of molecular assisted breeding, in particular to a PCR primer set for identifying or screening cabbage hybrid lethal parent type 2 and its application. The PCR primer set includes primers KBoHL2-X, KBoHL2-Y, and KBoHL2-Common. The present invention detects whether the genotype at positions 46, 333, and 244 of the C04 chromosome of the cabbage to be tested is T / T or C / C, and determines whether the cabbage has the characteristics of hybrid lethal parent type 2 according to the genotype of the cabbage to be tested: T / T genotype of the cabbage to be tested It is the hybrid lethal parent type 2, and the tested cabbage of the C / C genotype is not the hybrid lethal parent type 2. Using the PCR primers, kit or method of the present invention for assisted breeding has the advantages of simple operation, strong specificity, good stability, and early breeding can be realized, and the coincidence rate of identification results and field phenotypes reaches 100%. It has great application prospect.
Description
technical field [0001] The invention belongs to the technical fields of molecular biology and plant genetics and breeding, and relates to a PCR primer set for identifying or screening dead parent type 2 of cabbage hybrids and an application thereof. Background technique [0002] Heading cabbage, referred to as cabbage, belongs to the Brassica cabbage species of the Cruciferae (Cruciferae). Cabbage has strong adaptability and stress resistance, and has high nutritional and health value, so it is widely cultivated in China and even all over the world. Hybrid lethality refers to the phenomenon that the parents grow normally, but the hybrids appear wilting, yellowing, dwarfed plants, or even die. Hybrid lethality is an important reproductive isolation method, which is very important for the formation of species and the maintenance of species purity. In recent years, there have been more and more studies on the lethality of plant hybrids, but in cruciferous crops, most of them a...
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