Chinese cabbage est‑ssr marker and its primers and applications

A Chinese cabbage, labeling technology, applied in the field of molecular biology, can solve the problem that the Chinese cabbage EST-SSR primer is not saturated

Active Publication Date: 2017-09-15
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the development of Chinese cabbage EST-SSR primers has not yet reached saturation

Method used

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  • Chinese cabbage est‑ssr marker and its primers and applications
  • Chinese cabbage est‑ssr marker and its primers and applications
  • Chinese cabbage est‑ssr marker and its primers and applications

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1. Development of EST-SSR primers

[0039] The EST sequences used to develop EST-SSR primers came from the high-throughput transcriptome sequencing of Chinese cabbage (B.rapassp.pekinensis) rosette leaves and ball leaves by our research group (Wang et al., 2012). Through transcriptome sequencing, a total of 53.47 million high-quality EST sequences were obtained. Trinity software was used for these EST sequences ( http: / / trinityrnaseq.sourceforge.net / ) were assembled to obtain the corresponding contig and unigene, and then the TGICL software package (TIGR Gene Indices Clustering) was used to remove redundancy, and a total of 52,000 unigenes were obtained. In this study, the MISA software (http: / / www.pgrc.ipk-gatersleben.de / misa) was used to analyze and identify the EST-SSR of these 52,000 Unigenes (standards refer to Zhang et al., 2004), and a total of 10,400 EST-SSRs. Through the Chinese cabbage SSR database ( http: / / oilcrops.info / SSRdb ), a total of 27...

Embodiment 2

[0043] Application of embodiment 2.EST-SSR labeled primers

[0044] 2.1 Extraction of plant material and template DNA

[0045] Select 24 varieties (lines) of Chinese cabbage: 682, Guangdong Zao, Zaohuangbai, Z61-8, Fushan Baotou, Li-3, 212-7, Tianjin Qingmaye, Kuaicai No. 6, Golden Cabbage J, Siji Chinese cabbage, Siji Huangyang Xiaobaicai, Pinzao No. 1, Hanyu's special selection of yellow heart, Almighty Four Seasons Express, Jingyou Xiaobaicai Express, Korean Golden Baby, Keyi Xia Baby, Jinnuo Chunqiu Baby Cai, Four Seasons Green Stem small bitter vegetables, Oily Green 157, Tuyao rape, Degao Youliang green stems, Qingxiu F1 green stems. Marked as 1-24 groups respectively.

[0046] The total DNA was extracted by the modified CTAB method, and the specific steps were as follows:

[0047] 1. Take 1-10g of fresh Chinese cabbage leaves and quickly grind them into powder in liquid nitrogen;

[0048] 2. Transfer the frozen powder into a pre-cooled centrifuge tube, add an equal ...

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Abstract

The invention relates to Chinese cabbage EST-SSR (expressed sequence tag-simple sequence repeat) markers as well as primers and applications thereof. The invention discloses a Chinese cabbage EST-SSR marker BR-es10 and a primer thereof, a Chinese cabbage EST-SSR marker BR-es13 and a primer thereof, and a Chinese cabbage EST-SSR marker BR-es15 and a primer thereof. By carrying out amplification on the DNA of 24 Chinese cabbage varieties, and carrying out detection by using 6% denaturing polyacrylamide gel and / or 12% non-denaturing polyacrylamide gel electrophoresis, a situation that the amplified products of the three sets of EST-SSR primers are obvious in banding pattern, and good diversity is achieved among varieties is found, so that different varieties can be distinguished well, therefore, the markers and primers thereof can be applied to the identification of Chinese cabbage varieties and the construction of variety fingerprints.

Description

technical field [0001] The patent of the invention relates to Chinese cabbage EST-SSR markers and their primers and applications, in particular to three groups of Chinese cabbage EST-SSR markers and their primers and their application in the construction of variety fingerprints, belonging to the technical field of molecular biology. Background technique [0002] The emergence of molecular markers has transformed the selection in breeding work from traditional phenotypic selection to genotype selection, greatly improving the accuracy and efficiency of selection. Therefore, molecular markers have become an important and indispensable part of breeding work and genetics research since the emergence of molecular markers. Tool of. Especially the emergence of PCR-based molecular markers such as RAPD (Williams et al., 1990), SSR (Akkaya et al., 1992), AFLP (Vos et al., 1995), etc., because of their simplicity, rapidity and high efficiency. , so that the development and utilization ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李景娟高建伟王凤德李化银张一卉丁谦
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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