Co-segregation SSR (Simple Sequence Repeat) codominant molecular marker of onion male sterility gene Ms site and application thereof

Abstract

The invention discloses a co-segregation SSR (Simple Sequence Repeat) codominant molecular marker WH-SSR-1 of an onion male sterility gene Ms site. Nucleotide sequences of primers used for developing the marker are shown as SEQ ID NO. 3 and SEQ ID NO. 4, and nucleotide sequences of primers used for detecting an onion sample by applying the marker are shown as SEQ ID NO. 1 and SEQ ID NO. 2. The co-segregation SSR codominant molecular marker of the onion male sterility gene Ms site and the primers can be used for identifying the genotype of the onion male sterility gene Ms site by the followingsteps of: extracting leaf total DNA of the onion sample to be detected, amplifying with the primers through PCR, and detecting the size of the amplified product. By adopting the molecular marker provided by the invention, defects of the traditional molecular marker are overcome, a fussy screening process in the traditional method is avoided, and 4-6 years for crossing and test crossing as well asa great deal of manpower and material resource consumption in the process can be saved, therefore, the operation process is simpler and more efficient.

Description

technical field [0001] The invention relates to an SSR co-dominant molecular marker for co-separation of onion male sterility gene Ms sites. The development of the marker can be directly applied to onion molecular marker assisted breeding and belongs to the field of biotechnology. Background technique [0002] Onion (Allium cepa.L), also known as onion, onion, Liliaceae, Allium, biennial plant, has a cultivation history of more than 5,000 years. According to the 2009 FAO statistics, at least 175 countries cultivate onions. The world's onion planting area is 3.7 million hectares, with an output of 72.3 million tons. Among all vegetable crops, its planting area ranks second, and its output ranks third. Among them, my country has the world's largest onion production area and output, with a production area of ​​more than 1 million hectares, with a total output of 21.07 million tons, accounting for about 30% of the world's output. Onion has the reputation of health food. It can ...

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Problems solved by technology

and Havey in two open-pollinated onion populations[ AF, J McCallum, Y Sato and MJ Havey. Molecular tagging of the Ms locus in onion. Journal of the American Society for Horticultural Science, 2002, 127, 576-582.], using AOB272 (0.9cM) as an auxiliary tool, Tried to select the maintainer line, but found that the marker has little association with the maintainer line. The author thinks that it may be because in the long-term open pollination environment, sufficient chromosome exchange between the AOB272 marker and the Ms locus has been carried out, so only this molecule is used markers cannot accurately select the maintainer line

In our previous study, a cDNA molecular marker (WHR240) related to the male sterility restorer gene was developed, and this marker was verified in many onion materials, but because the marker was reverse transcribed The cDNA of a specific period and a specific tissue is used as a template, so it has certain limitations in operation and specific utilization (prerequisites such as RNA extraction, reverse transcription, and single dominant marker), and it is still difficult to identify large-scale breeding materials. there are certain limitations

In addition, the Onion, Ginger, and Garlic Research Center of Vegetable Research Institute of Shandong Academy of Agricultural Sciences (hereinafter referred to as: the research center) has made a breakthrough in the molecular markers of onion male sterility gene loci, and has developed the onion male sterility gene ms and The fertility restorer gene Ms is closely linked co-segregated DNA molecular markers, which can accurately determine whether Ms or ms genes are pure, but because they are not co-dominant markers, they cannot be amplified once to complete the genotype judgment, and must be separated After amplifying with the sterility gene linkage marker and the fertility restoration gene marker, the genotype is judged comprehensively based on the results obtained twice, and there is also the possibility of misjudgment of the amplification failure

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