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Molecular marker closely linked with cabbage type rape seed coat color and application of molecular marker

A Brassica napus and molecular marker technology, applied in the field of plant molecular genetics, can solve problems such as the lack of versatility of linkage markers, the lack of a clear genetic model, and the complex genetic mechanism of yellow seed traits in Brassica napus

Active Publication Date: 2015-06-24
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a lot of research has been carried out on the mapping of yellow seed traits in Brassica napus, and some molecular markers closely linked to yellow seed traits have been detected and converted into SCAR markers, due to the different sources of yellow seed genes in Brassica napus and the existence of polygenic inheritance , allotetraploidy, maternal effect, easily affected by many factors such as environmental conditions (Deynze et al., 1995; Shirzadegan, 1986), so far no QTL for related traits has been cloned in Brassica napus
Therefore, the genetic mechanism of the yellow seed trait in Brassica napus is relatively complicated, and there is no clear genetic model so far, and its linkage markers also lack certain versatility, which limits the assisted selection in breeding.

Method used

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  • Molecular marker closely linked with cabbage type rape seed coat color and application of molecular marker
  • Molecular marker closely linked with cabbage type rape seed coat color and application of molecular marker
  • Molecular marker closely linked with cabbage type rape seed coat color and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Construction and Character Measurement of Brassica napus Recombinant Inbred Line Mapping Population

[0034] The mapping population used in this example is a recombinant inbred line population constructed by continuous selfing of the brassica napus yellow seed line GH06 and black seed line ZY821 by continuous selfing. The phenotypic data of seven different environments of this group were obtained by using a near-infrared analyzer, and the analysis of the data by the analysis software SPSS 13.0 showed that the color of the testa in different environments showed a continuous approximately normal distribution, with a wide range of variation. It is proved that the seed coat color of Brassica napus is a typical quantitative trait ( figure 1 ).

Embodiment 2

[0035] Example 2 Extracting Genomic DNA

[0036] According to the CTAB method of Doyle (1990) et al., the CTAB method was slightly improved, and the genomic DNA of the leaves of the two parents of Brassica napus GH06 and ZY821 and the recombinant inbred line population were extracted. The specific operation steps are as follows:

[0037] A) Put about 0.1g of fresh leaves into a 2ml centrifuge tube, and put 4 steel balls to crush the leaf tissue.

[0038] B) To prevent oxidation during pulverization, 100 μL of 2×CTAB extraction buffer containing 2% β-ME (mercaptoethanol) was added.

[0039] C) Put the 2ml centrifuge tube filled with materials into SPEX GENO 2010 in the United States A high-throughput tissue grinder pulverizes leaf tissue at a speed of 1250 rpm for 55 s.

[0040] D) After the pulverization is completed, add 700 μL of CTAB extraction buffer preheated at 65° C., shake vigorously to make it mix quickly, and then keep it at 65° C. for 45 minutes, shaking 3-4 time...

Embodiment 3

[0045] Example 3 Construction of Molecular Marker-Linked Genetic Map of Brassica napus and Mapping of QTL for Seed Coat Color

[0046] Four different markers, SSR, SRAP, RAPD and IBP, were used to construct the genetic linkage map of molecular markers in Brassica napus. The SSR markers mainly include: Na-, Ol- published by BrassicaDB (http: / / www.ukcrop.net) , Ra-, Ni-primers, a total of 284 pairs of primers; Piquemal et al. (2005) published a total of 157 pairs of CB-, BRAS-, MR- and MD primers; a total of BRMS-primed 130 pairs (Fu et al., 2007); 97 pairs of FITO-primers published by Osborn Laboratory; 109 pairs of sR, sN, sO, nga-, BN-primers published by NAFF in Canada; Dr. Beom-Seok Park of Korea A total of 240 pairs of KS-, H-, B- and S-primers donated by Dr. Soo-Jin Kwon (http: / / brassicadb.org / brad); BnGMS-primers published by Cheng et al. (2009) 135 pairs; a total of 124 pairs of EJU, ENA, GOL, niab- and cnu-primers published by Choi et al. (2007); and SWU-specific prim...

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Abstract

The invention belongs to the technical field of plant molecular genetics, and in particular relates to a molecular marker closely linked with a cabbage type rape seed coat color and an application of the molecular marker. The invention aims to provide a new choice for fine positioning and cloning of color main effect QTL in the future. The invention discloses a molecular marker SWUA09-355 closely linked with the cabbage type rape seed coat color, and has nucleotide sequences shown by SEQ ID NO.1 and SEQ ID NO.2. The invention also provides a primer for amplifying the molecular marker and a method for obtaining the molecular marker. The invention localizes an SSR molecular marker SWUA09-355 closely linked with seed coat color properties, and the main effect QTL can explain 41.38-64.17% of phenotypic variations in different environments.

Description

technical field [0001] The invention belongs to the technical field of plant molecular genetics, and specifically relates to a molecular marker closely linked with the color of the brassica napus seed coat and its application. Background technique [0002] Brassica napus belongs to the cruciferous family and occupies an important position in the world's oil crops. It is the third largest source of vegetable oil after soybean oil and palm oil, and is also one of the important sources of plant protein feed. In rapeseed genetics and breeding, the reason why the yellow seed trait has attracted great attention from breeders at home and abroad is mainly because under the same genetic background, compared with black seeds, yellow seeded rapeseed lines have thinner seed coats, oily It has a series of advantages such as high protein content, low hull rate, lignin, cellulose and polyphenol content, and good quality (Rahman et al., 2001; Theander et al., 1977). The results of the stud...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 曲存民李加纳卢坤付福友王瑞徐新福张凯
Owner SOUTHWEST UNIVERSITY
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