Transformation Of Soybean Cryptochrome Pigment Gene Into Potato And Its Functional Analysis

Cryptochrome is a kind of blue light receptor.It is mainly responsible for the external light environment in plants.It has many biological functions and plays an important regulatory role in plant growth and development,such as response to stress,photomorphogenesis,Directional growth,stomatal opening and closing,flowering time,root growth,and shading response.The cryptochrome-related gene has great potential for development as a genetic resource for molecularly improving the agronomic traits and stress resistance of crops.At present,the research on the function of cryptochrome pigment gene is only concentrated in a few plants such as Arabidopsis,and the research and utilization in potato and other economic plants are very limited.Therefore,genetic engineering technology is used to transform cryptochrome-related genes into potato,which provides an important research method for creating excellent germplasm materials for potato.Based on the high-efficiency genetic transformation system of potato established by Ningxia Agricultural Biotechnology Key Laboratory,the soybean cryptochrome gene was transformed into potato,and the molecular identification and phenotypic identification were used to screen out the sensitivity to photoperiod change and drought resistance and tolerance.The salt-enhanced transgenic lines can reveal the function of cryptochrome gene after transforming potato,and can show more sensitivity to light reaction and strong drought resistance and salt tolerance.The main findings are as follows:1.Agrobacterium-mediated method,using the potato variety "Atlantic" as a receptor,five related genes of soybean cryptochrome(pearlygate104-Glyma.10G180600,pearlygate104-Glyma.04 G101500,pearlygate 104-Glyma.06G 103200,pearlygate 104-Glyma.02G005700,peary gate 104-Glyma.20 G209900)were transformed,and 25 positive strains were obtained by PCR and RT-PCR.2.The artificial simulated lighting conditions,long daylight(light 16 h/dark 8 h),middle sunshine(light 12 h/dark 12 h)and short sunshine(dark 16 h/light 8 h)three kinds of photoperiod treatment The effect of transforming the growth of soybean cryptochromin gene positive strains.The results showed that with the increase of illumination time,the plant height of the transgenic lines decreased significantly,and the number of leaves,leaf area and stem diameter increased significantly.Under short-day conditions,the plant height,leaf number,stem diameter and leaf area of the transgenic lines increased by 1-5.5 cm,2-16,0.26-0.79 mm and 0.15-0.46 cm2 compared with the control;in the case of medium-day sunshine,transgenic The plant height,leaf number,stem diameter and leaf area of the lines increased by 1.5-4.5cm,2-18 pieces,0.26-0.8mm and 0.4-2.46cm2 compared with the control;under long daylight conditions,the plant height of the transgenic lines decreased compared with the control.0.5-4.5cm,the number of leaves,stem diameter and leaf area of transgenic lines increased by 6-27 pieces,0.31-2.38mm and 0.88-3.41cm2 respectively compared with the control.It is indicated that the conversion of soybean cryptochrome pigment gene significantly improves the ability of potato to respond to light signals.3.Using 120 mmoL/L NaCl and 10%PEG to simulate salt and drought stress respectively,the plant height and root length of transgenic lines were determined by physiological indexes related to drought resistance and salt tolerance.The results showed that:drought stress conditions Under the condition of salt stress,the plant height and root length of the transgenic plants increased by 0.5-4.0 cm and 1.8-4.1 cm,respectively.The content of proline under drought stress increased by 123.8?g/g and 285.2?g/g,respectively.The content of malondialdehyde under salt and drought stress decreased by 14.8nmol/ml and 34.2 nmol/ml,respectively.The test results further confirmed that the transfer of cryptochrome gene can significantly improve the drought and salt tolerance of the potato.4.A full-length cDNA of Cryptochrome 1 was cloned from the leaves of potato cultivar "Atlantic" by RT-PCR.The expression analysis of the gene tissue by qPCR showed that StCRYl gene was expressed in potato roots,stems,leaves,flowers and tubers,and the expression of this gene was highest in leaves.In summary,this study used Agrobacterium-mediated transformation of soybean cryptochrome pigment-related genes into potato to obtain transgenic positive strains;transgenic positive strains were lighted by different photoperiod and drought and salt stress treatments.Reaction,drought resistance and salt tolerance studies.In addition,the cryptic pigment gene StCRY1 was isolated from the potato cultivar"Atlantic" by RT-PCR method,and the expression pattern of the gene was analyzed.The biological function of potato StCRYl was preliminarily clarified,and the function of plant CRY1 gene was enriched.Provide support for the cultivation of new varieties of resistant potato.