The Impact Of High Temperature Stress On Physiological-biochemical Characteristic Of Capparis Spinosa L. Seedlings And Screening Of The Differentially Expressed Genes

Global warming not only causes serious problems such as high temperature stress but also couldthreaten to agricultural production on our planet, It’s an important branch of modern plant breedingprogram on stress resistance to enhance or improve the capability of high temperature tolerance in plant.Capparis spinosa (Capparis spinosa L.) is a typical arid area of desert plants, it is extremely resistant todrought and high temperature, is the study of plant resistance rare material, also can be used as a donor ofhigh temperature resistant gene, and provide excellent resistance gene resources for breeding. But theCapparis spinosa seed is hard to sprout, so we firstly examined the Capparis spinosa seed germination; wenext sought to determine whether effected physiological and biochemical indexes of seedlings the hightemperature stress; at last, construct the high quality RNA extraction and the optimal cDNA-SRAP reactionsystem, were normal and high temperature stress of Capparis spinosa differential expression gene bybioinformatics method, functional analysis and classification of the differentially expressed genes, in orderto more clearly understand the causes of Capparis spinosa high temperature tolerance, to further tap theCapparis spinosa lay the foundation of high temperature resistant gene, and laid the theoretical foundationfor improving the stress resistance of plants. The results are as follows:（1）Capparis spinosa seed germination time was20d, cut the seed treatment germination rate was44, notthrough the seed germination rate was only5, the difference was significant (p<0.01). With the camerapointing broken seed treatment can significantly improve the germination rate (p<0.01), which pierced theseed coat and200mg/L GA3combination treatment germination rate reached70. The same physical andchemical treatment, that treatment can significantly improve the germination rate (p<0.01), and does notaffect the root length, shoot length and leaf length(p<0.05).（2）Capparis spinosa seedlings compared with the control under high temperature stress, the prolinecontent, MDA content, electrical conductivity were increased with the prolonging of stress time andtemperature increased; the soluble protein content decreased with increase of the extension of the stresstime and temperature; peroxidase (POD) activity, superoxide dismutase enzyme (SOD) activity with theincrease of stress time and temperature increased first and then decreased; the membrane system after along time of high temperature stress of Capparis spinosa seedlings were damaged, the level of membranelipid peroxidation increased, the ability of osmotic adjustment and antioxidant system of protective enzymeactivity also decreased significantly, especially the temperature at more than40℃temperature and thestress time more than5days, Capparis spinosa seedlings from heat harm is more serious.（3）RNAprep Pure Plant Kit method is the best method of acupuncture effect Capparis spinosa extracttotal RNA, using the method of orthogonal design and single factor combination of Capparis spinosa,eventually established the optimal SRAP-PCR reaction system: the total volume of25μL, template cDNA40ng, dNTPs0.2mmol/L,TaqDNA polymerase0.75U, primers0.2μmol/L, Mg2+1.35mmol/L.（4）In this study, we performed PCR using30pairs of primers and amplified334differential bands, theselection of the103stable expression of differential bands were sequenced by NCBI Blast, comparison,homologous fragments of47annotated genes, accounting for about45.6of the measured number, andanother56fragments for unknown gene, may be functional genes yet to be discovered. shorter or yet to be discovered. The sequence of gene function prediction, its function can be divided into five categories, thefirst is related to transport protein (10,9.7%), such as grid protein, binding protein and membrane protein;The second category in adversity stress related protein (13,12.6%), such as protein phosphatase,cytoplasmic ascorbic acid salt, peroxidase and so on; The third class is related to signal transduction protein(15,14.6%), such as serine/threonine protein kinase, F-box protein, etc.; The fourth class is assumedprotein (9,8.7%); Class5no homologous sequences (56,54.4%), protein may be a potential function.