A Chinese cabbage burr indel marker and its detection primer and application
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A technology for detecting primers and Chinese cabbage, which is applied in the field of molecular biology and can solve problems such as lack of markers and hindering breeding.
Active Publication Date: 2022-03-18
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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But so far, the lack of markers closely linked to the burr trait has seriously hindered the breeding process, which means that the development of molecular markers closely linked to the burr trait of Chinese cabbage will be of great help to the rapid breeding of new varieties of hairless and disease-resistant Chinese cabbage. Has important guiding and practical significance
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Embodiment 1
[0039] Example 1. Development of InDel labeled primers
[0040] Taking Chinese cabbage burr formation-related genes as candidate genes, hairy (G291) and hairless (ZHB) Chinese cabbage genome-wide resequencing was performed using resequencing technology. According to the resequencing results, the candidate genes in hairy and hairless Chinese cabbage were compared, and primers were designed for one of the InDel marker TR-1. The sequence comparison of the Chinese cabbage InDel marker TR-1 is shown in figure 1 shown. The area marked by a line in the figure is the InDel marker TR-1.
[0041] Hairy Chinese cabbage (G291) has a 12 bp DNA insertion compared to hairless Chinese cabbage (ZHB). The detection primers for Chinese cabbage InDel-labeled TR-1 were designed, and the primer sequences are shown in SEQ ID NO.1 and SEQ ID NO.2.
Embodiment 2
[0042]Application of embodiment 2.InDel labeled primers
[0043] 2.1 Extraction of plant material and template DNA
[0044] Select the first generation of Chinese cabbage hybrid commercial varieties Qingyan Chunbai 2, Degao Baihe, Xibai 45, Xibai 8, Zhonghua 1, Qingyan CR20, Qingyan Xiabai 2, Zhonghua 1, Gaokang 2 , Xibai 5, Xiayou 2, Jintongyunv, Gongguan 2, Fengkang 60, Huayangbai, Degaoxikang 65 and Chinese cabbage inbred lines Guangdong Zao, Y2, L41, L46, AM4-1, AM3 -1, AM5-2, AM9-1, AM94, AM30, AM54, AM72-1, AM10-1, AM56-1, and hybrid F2 leaves of ZHB and G291 were used as materials.
[0045] The total DNA was extracted by the modified CTAB method, and the specific steps were as follows:
[0046] 1. Take 1-10g of fresh Chinese cabbage leaves and quickly grind them into powder in liquid nitrogen;
[0047] 2. Transfer the frozen powder into a pre-cooled centrifuge tube, add an equal volume (w / v, g / ml) of 2×CTAB extraction buffer, and incubate at 65°C for 20 minutes;
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Abstract
The patent of the present invention relates to the detection primer labeled with InDel and its application in the breeding of burr-free Chinese cabbage. The detection primer of Chinese cabbage InDel-labeled TR-1, the detection primer is a pair, the nucleotide sequence of the upstream primer is shown in SEQ ID NO.1, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.2. The invention discloses for the first time the InDel-marked TR-1 related to the burr of Chinese cabbage and its detection primers. PCR amplification is carried out on the DNA of hairy and hairless Chinese cabbage inbred lines and the F2 generation isolated population material generated by the hybridization of the two. It was detected by 8% denaturing polyacrylamide gel electrophoresis. It was found that the InDel marker TR-1 related to the burr of Chinese cabbage can clearly distinguish between hairy and hairless Chinese cabbage varieties, and can be used for the identification of new hairless Chinese cabbage varieties. breeding.
Description
technical field [0001] The invention relates to an InDel label of Chinese cabbage burrs, detection primers and application thereof, and belongs to the technical field of molecular biology. Background technique [0002] Molecular marker-assisted selection (marker-assisted selection, MAS) is not limited by environmental conditions, and can be carried out at the level of DNA molecules at any growth stage of plant development (Tanksley, 1989). It overcomes the shortcomings of traditional plant breeding such as long time, slow speed, uncertainty, and environmental conditions. At present, a variety of plant molecular marker technology systems have been established in the world and applied to the genetic improvement of crops, which has opened up a new way for crop breeding. The use of molecular marker technology for rapid screening, especially the emergence of molecular markers, has transformed the selection in breeding from traditional phenotype selection to genotype selection, g...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王红霞李景娟杨晓刚丁谦李化银王凤德张一卉高建伟
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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