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Molecular markers, primer pairs, and molecular marker methods and applications for identifying early and late flowering of head cabbage

A technology of molecular markers and cabbage, applied in the field of molecular biology, can solve the problems of heavy workload, influence by genetic factors, cumbersome operating procedures, etc., and achieve the effect of overcoming the long time period

Active Publication Date: 2020-11-24
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are mainly two methods for screening breeding materials. One is the traditional breeding method. This method requires the selection and identification of bolting resistance traits across the year. The cycle is long, the operation procedures are cumbersome, and the workload is heavy. In addition to being affected by genetic factors, it is also closely related to environmental factors, so relying on phenotypic identification has greater inaccuracy; another method is the molecular marker-assisted method developed in recent years, which combines molecular marker technology with traditional breeding. Combining, using stable and reliable DNA molecular markers to replace environmental sensitive phenotypic markers to screen breeding materials

Method used

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  • Molecular markers, primer pairs, and molecular marker methods and applications for identifying early and late flowering of head cabbage
  • Molecular markers, primer pairs, and molecular marker methods and applications for identifying early and late flowering of head cabbage
  • Molecular markers, primer pairs, and molecular marker methods and applications for identifying early and late flowering of head cabbage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The statistics of embodiment 1 early flowering head cabbage and late flowering head cabbage

[0049] Field flowering time investigation of different cabbage, 26 early-flowering cabbage and 24 late-flowering cabbage were screened out. The flowering time of early-flowering and late-flowering cabbage was counted for three consecutive years. The results are as follows: the average flowering time of early-flowering cabbage in 2015 was 193.23±8.10 days, and the average flowering time of late-flowering cabbage was 203.5±9.57 days. The flowering time of cabbage flowering was about 10 days later than that of early flowering cabbage; the average flowering time of early flowering cabbage in 2016 was 199.77±8.32 days, and the average flowering time of late flowering cabbage was 215.96±7.77 days, and the average flowering time of late flowering cabbage was 215.96±7.77 days. Heading cabbage flowered earlier The delayed flowering time of cabbage was about 16 days; the average flowerin...

Embodiment 2

[0050] The extraction of embodiment 2 cabbage genomic DNA

[0051] (1) Get 50 parts of young leaves of head cabbage screened in Example 1, store them in 1.5mL centrifuge tubes respectively, and grind them into pulverized shapes in liquid nitrogen;

[0052] (2) Add a mixture prepared by mixing 700 μl CTAB extract and 14 μl β-mercaptoethanol at 65°C to each centrifuge tube containing the sample, bathe in 65°C water for 45 minutes, and mix 3-4 times in between;

[0053] (3) Add an equal volume of the mixed solution obtained in step (2) to each centrifuge tube with the sample in the chloroform / isoamyl alcohol mixed solution with a volume ratio of 24:1, shake gently, mix well, emulsify for 10min, and centrifuge at 10000rpm 10min, take the supernatant in a new centrifuge tube;

[0054] (4) in the new centrifuge tube that step (3) gained supernatant is housed, add the chloroform / isoamyl alcohol mixed solution that is 24:1 with the volume ratio of the supernatant equal volume of step...

Embodiment 3

[0058] Example 3 Anchoring of BoFLC2 Gene Differential Fragments

[0059] (1) According to the BoFLC2 reference genome sequence, use Primer3 online primer design software to design primer F11 (Forward primer: 5'-ATTCGCCCACCGATTCGTAT-3', Reversed primer 5'-GCCGGCGGAGAAGTTGTATA-3'), 20μl reaction system (ddH 2 O: 4.2μl; 2×Phanta Max buffer: 10μl; dNTPMix (10mM each): 0.4μl; Primer F (10μM): 1μl; Primer R (10μM): 1μl; Phanta maxsuper-fidelity DNAPolymerase (1U / μl): 0.4 μl; DNA (100ng / μl): 3μl), using PCR reaction program (98°C 30s, {98°C 15s, 62°C 15s, 72°C 3min}×7cycles, {98°C 15s, 55°C 15s, 72°C 3min} ×22cycles, 10min at 72°C, stored at 16°C), amplified the BoFLC2 gene promoter region, and imaged by 1% agarose gel electrophoresis, the results are shown in figure 1 , the size of the amplified fragment in the promoter region of early-flowering and late-flowering cabbage was about 2.0 kb, and there was no difference in size;

[0060] (2) According to the BoFLC2 reference genome ...

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Abstract

The invention belongs to the field of molecular biology, and particularly relates to a molecular marker, a primer pair and a molecular marking method used for identifying the flowering time of brassica oleracea and application of the molecular marker, the primer pair and the molecular marking method. The molecular marker and the primer pair can be used for quick identification of a brassica oleracea breeding material and variety which are resistant to bolting and flower late and for the marker assisted selection of transfer breeding of the material which is resistant to bolting and flowers late, and are of great significance in brassica-oleracea-like vegetable breeding practice and theoretical research on bolting resistance.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to a molecular marker, a primer pair, a molecular marker method and an application for identifying the flowering time of head cabbage. Background technique [0002] Heading cabbage (Brassica oleracea var.capitata), hereinafter referred to as cabbage, is a biennial vegetable crop of the genus Brassica in the family Brassicaceae. The process of vernalization in winter, flowering and fruiting in spring and summer of the second year, complete reproductive growth. The edible organs of head cabbage are mainly leaf bulbs, but in the production of head cabbage, due to the poor bolting tolerance of cabbage varieties and the frequent coldness in spring, head cabbage experiences low temperature vernalization before the leaf bulbs mature. The immature bolting phenomenon, once bolting occurs, will prevent the growth of leaf bulbs and directly bolt and bloom, and enter the reproductiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 宋洪元李勤菲李樟萍任雪松司军
Owner SOUTHWEST UNIV