Method for screening real-time fluorescent quantitative PCR internal reference molecules of desert plant Eremosparton songoricum (Litv.) Vass.

Abstract

The invention relates to a method for screening real-time fluorescence quantitative PCR internal reference molecules of a desert plant Eremosparton songoricum (Litv.) Vass.. The invention is characterized in that homologous clone is used for cloning 8 internal reference molecules in Eremosparton songoricum (Litv.) Vass., and a real-time fluorescent quantitative PCR internal reference gene specific primer is designed, 10 types of seedling grown for two weeks with abiotic stress and non stress (contrast) are taken as the experimental materials for carrying out a fluorescent quantitative PCR experiment to verify, analysis of fluorescent quantitative data is carried out by using geNorm software, so that the optimum and the stablest internal reference molecules screened for fluorescent quantitative researches under various abiotic stresses developed by the seedling of Eremosparton songoricum (Litv.) Vass. are EF and alpha-tubulin. The method of the invention is capable of avoiding the errors on RNA quality, output, inverse transcription efficiency and the like of the seeding material of Eremosparton songoricum (Litv.) Vass. under different abiotic stresses, better correcting and standardizing the data obtained by the real-time fluorescent quantitative determination and enhancing the accuracy and reliability of researches.

Description

technical field [0001] The invention relates to a method for screening internal reference molecules suitable for the real-time fluorescent quantitative PCR research of desert plant Junggar leafless bean seedlings under various abiotic stresses Background technique [0002] With global warming and the intensification of the greenhouse effect, agricultural production in arid areas is facing severe challenges. Facing the grim reality of my country's growing population, dwindling arable land and water and fertilizer resources, and increasingly sharp contradictions between food supply and demand, how to quickly increase crop yields while saving resources and improving the environment has become a major demand for the sustainable and efficient development of my country's agriculture. The drought-resistant gene resources accumulated in the long-term evolution process in arid areas are the source of germplasm innovation! [0003] The harsh natural environment in the arid region has...

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Problems solved by technology

[0008] In recent years, some stress-resistant genes have been cloned from Junggar leafless bean, such as the dehydration response element binding protein (DREB) transcription factor gene (NCBI accession number: HQ687367). The characteristics and functions of these genes need to be studied urgently, but about the The sequence data of the internal reference gene has not been reported, and the research on the most suitable internal reference molecule is even blank

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