Double-dominant marker for accurately authenticating S/N cytoplasm genotype of onions and application of double dominant marker

A cytoplasmic and genotypic technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems affecting the selection efficiency, lack and other problems, so as to improve the selection efficiency, save time and cost, and ensure accuracy. The effect of reliability

Active Publication Date: 2018-04-06
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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Problems solved by technology

Although this marker can effectively avoid misclassification of cytoplasmic types due to experimental manipulation errors and ineffective PCR amplification, due to the lack of stable N-type cytoplasmic specific markers, it is impossible t

Method used

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  • Double-dominant marker for accurately authenticating S/N cytoplasm genotype of onions and application of double dominant marker
  • Double-dominant marker for accurately authenticating S/N cytoplasm genotype of onions and application of double dominant marker
  • Double-dominant marker for accurately authenticating S/N cytoplasm genotype of onions and application of double dominant marker

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Embodiment 1

[0038] (1) Materials and methods

[0039] 1. DNA and RNA extraction and detection

[0040] For the extraction method of onion cytoplasmic male sterile line and its corresponding maintainer line genomic DNA, refer to the instructions of the quick plant genome extraction kit produced by Beijing Tiangen Biochemical Technology Co., Ltd., and for the extraction of total RNA, refer to the kit produced by Beijing Tiangen Biochemical Technology Co., Ltd. RNAprep Pure plant total RNA extraction kit instructions, agarose gel electrophoresis and spectrophotometer to detect the quality of DNA and RNA.

[0041] 2. Acquisition of differentially expressed fragments

[0042] In the published complete mitochondrial genome sequence of onion CMS-S (GenBank accession: KU318712), 58 new ORFs were found. The inventors used the cDNA of the onion sterile line S118 and the maintainer line N218 as templates to analyze the 58 ORFs Differential expression analysis was carried out, and a differentially ...

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Abstract

The invention discloses a double-dominant marker for accurately authenticating an S/N cytoplasm genotype of onions and application of the double-dominant marker. The length of a specific fragment of an S-form cytoplasm in the double-dominant marker is 232bp, and the length of a specific fragment of an N-form cytoplasm is 196bp. The invention further discloses SCAR marking primers SEQ ID NO.5 and SEQ ID NO.6. A prepared SCAR molecular marker is utilized for rapidly and accurately judging cytoplasm types of onion materials, so that the misclassification of cytoplasm genotypes caused due to experimental operate miss and invalid PCR amplification can be avoided; and furthermore, DNA can be extracted from mixed group samples when a maintainer line is selected in a large group and can be used for primarily screening whether a maintainer line single plant exists, so that the selection efficiency is greatly improved, the time and the cost are saved, and the double-dominant marker has importantsignificances to the matching of a breeding sterile line and a maintainer line of the onions and the breeding of hybrid varieties of the onions.

Description

technical field [0001] The invention belongs to the technical field of plant breeding, relates to a molecular marker for onion breeding and its application, in particular to a double-dominant molecular marker for identifying the S / N cytoplasmic genotype of onion and its application, which can be applied to molecular marker-assisted breeding Accurate selection of onion male sterile lines and matching maintainer lines. Background technique [0002] Onion (Allium cepa.L), also known as bulb onion, round onion, jade onion, onion, Dutch onion, Piyazi, etc., is a biennial diploid plant belonging to the Liliaceae Allium genus. According to records, it has a cultivation history of more than 5,000 years , onion was introduced into my country in modern times, and it has only been cultivated in my country for more than 100 years. However, due to its strong adaptability and resistance to storage and transportation, although the history of onion planting in my country is short, its develo...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 杨妍妍吴雄刘冰江霍雨猛曹德高
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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