Momordica grosvenori gender gene SgACS2 and amplification method thereof
A Luo Han Guo and gender technology, applied in genetic engineering, plant genetic improvement, chemical instruments and methods, etc., can solve the problems of unidentifiable sex at the seedling stage, loose sex linkage, and affect the sex identification at the seedling stage, so as to accelerate the process of genetic improvement , saving land and improving breeding efficiency
Inactive Publication Date: 2018-10-09
广西作物遗传改良生物技术重点开放实验室
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Problems solved by technology
Although the successful popularization and application of tissue culture seedlings has rapidly expanded the scale of industrial planting, the problem of unidentified sex at the seedling stage has been seriously affecting the genetic improvement of Luo Han Guo varieties.
[0003] Existing random polymorphic DNA markers (Tao Li et al., 2005; Qin Xinmin et al., 2007; Wei Suling et al., 2008; Zhang Huixin et al., 2013) and isozyme markers (Li Huimin et al., 2007) is a non-sex-determining gene marker, which is not closely linked with sex and the marker is lost (Huang Zimei, 2007), which seriously affects its sex identification at seedling stage
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[0022] It should be noted that the experimental methods described in the following embodiments, unless otherwise specified, are conventional methods, and the reagents and materials, unless otherwise specified, can be obtained from commercial sources.
[0023] A method for amplifying the sex gene SgACS2 of Luo Han Guo, comprising the following steps:
[0024] Step 1. Collect the budding stems of Luo Han Guo, and extract the total RNA of Luo Han Guo with the plant total RNA extraction kit (DP432) of Tiangen;
[0025] Step 2. Using Clontech's The MMLV reverse transcriptase provided by RACE 5' / 3' Kit User Manual provided the total RNA obtained in reverse transcription step 1, and adapter primers were added during reverse transcription to construct 3'-RACE and 5'-RACE cDNA template libraries; Clontech's SeqAmp DNAPolymerase (Cat.No.638504) for amplification efficiency PCR amplification, 5'-RACE adapter primer and specific primer GST-F paired to obtain 5'-RACE-Ready cDNA, 3'-RACE...
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The invention discloses a momordica grosvenori gender gene SgACS2. A male strain has the gene sequence shown as SEQ ID No.1, a female strain has the gene sequence shown as SEQ ID No.2. The invention provides a method for PCR amplification detection on the gender of momordica grosvenori at a seedling stage, and also provides an important genetic sequence information basis for the genetic engineering breeding of momordica grosvenori and the development of a new method for identifying the gender at a seedling stage by specific polymorphic sites, thereby accelerating the process of genetic improvement of momordica grosvenori varieties, and the method has the beneficial effects of saving a lot of land, manpower and financial resources.
Description
technical field [0001] The invention relates to the field of plant molecular biology. More specifically, the present invention relates to a Luo Han Guo sex gene SgACS2 and its amplification method. Background technique [0002] Luo Han Guo (Siraitia grosvenorii) is a unique medicinal plant of Cucurbitaceae in my country. ), anti-cancer (Takasaki et al, 2003), etc. It is the first batch of Chinese medicine varieties listed by the Ministry of Health as both a drug and a food list, and it is also the most famous authentic medicinal material in Guangxi. The active ingredient of Luo Han Guo, glucoside V (Liu Ting et al., 2007), is one of the strongest non-sugar sweeteners in the world, with a sweetness 300-400 times that of sucrose (Matsumoto et al, 1990), and it is low in calories and non-toxic (Jin et al, 2007; Qin et al, 2006), is a natural sweetener suitable for people with diabetes and obesity. Since its successful entry into the US market (Rahul et al, 2013), the market d...
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IPC IPC(8): C12N15/29C12N15/10
CPCC07K14/415
Inventor 莫长明郭文锋李忠张玉
Owner 广西作物遗传改良生物技术重点开放实验室