Preliminary Verification Of Cold Resistance Of GPAT Gene In Lilium Formolongi

In this study,the PCR primers and hybridization probes were designed with the full-length sequence of the Lilium formolongi GPAT gene.The expression pattern of the GPAT gene is study by qRT-PCR technique in time and space.To study the transcriptional level of GPAT gene in different developmental stages of Lilium formolongi(seedling stage,flowering stage and green fruit stage),and to analyze the relationship between GPAT gene transcription level and growth development.According to the data of fatty acid composition and content,and the change of temperature,analyze the relationship between the transcription level of GPAT gene and the fatty acid composition and content.To further verify its time expression at the protein level by Western Blot technique,to reveal the main period of its physiological role.The specific test results are as follows:1.For different time at different temperatures on the whole plant in seedling stage,flowering stage,green fruit stage of Lilium formolongi,determination of fatty acids in Lily leaves by gas chromatography(Palmitic acid,palmitoleic acid,stearic acid,oleic acid,linoleic acid,linolenic acid),analysis of changes in unsaturated fatty acid content.Among them,the content of palmitic acid in saturated fatty acids is about 20%-30%of the total content,and the content of linolenic acid in unsaturated fatty acids is about 30%-50%of the total content.It has a great influence on the growth and development of lily at 4?.Under low temperature,linoleic acid and linolenic acid changed greatly,so as to improve plant cold resistance.2.On the whole plant in seedling stage,flowering stage,green fruit stage of Lilium formolongi leaves,roots and flowers were treated at different temperature and different time,using real-time fluorescence quantitative PCR technology,expression quantity of GPAT gene from lily.Data analysis show that in the leaves,roots and petals of these three parts of the organization,the expression of GPAT gene in leaves was the highest,which indicate that the expression of GPAT gene in green tissue was high,which was consistent with the gene locus on chloroplast.At seedling stage flowering stage and green fruit stage three development stages of lily,effects of low temperature at 4? on seedling stage and green fruit stage greatly.3.The effect of low temperature at 4? on the expression of GPAT protein in leaves of Lilium formolongi is study by western blot.The results showe that the low temperature at 4?has a great effect on seedling and green fruit,and has little effect on flowering lily.The expression level of GPAT protein decrease at 4h after seedling stage,indicating that the leaf is affect by the persecution of low temperature.The expression level of GPAT protein reach the maximum at 8h,and reached the normal expression level after 24h.The protein expression in the green fruit is lower than the normal level,and the results are consistent with the real-time quantitative PCR.Indicating that the expression level of GPAT gene in green fruit increased so as to improve plant cold resistance.