Analysis Of Differentially Expressed Genes At Maturation Stages Of High And Low Oil Content Soybean Seed

Soybean [Glycine max (L.)] is one of the most important sources for plant oil . Soybean originates form china, the breeding resources are abundant, the quality and heredity multiplicity are high. Now, the fatty acid metabolism mechanism aspect of soybean already had some research, but most of these research about fatty acid pathways just from the single genes, Few of them research the fatty acid synthesis metabolism network from the soybean seed growth. How to find the key factors from complex biochemistry system, expounds its biology function and transformation it according to human' needs were very important, It's very useful to improve its fatty acid composition. "zhongdou 32" and "youchun 02-6" were the soybean lines bred by China, the oil content is 22.48% vs 17.72%,the protein content is 37.07% vs 44.07% , The difference was obvious . Suppression subtractive hybridization (SSH) is a matured and high efficient tool to find the differentially expressed genes. Using SSH to compared with two soybean variety growth process in gene expression differences can search the key factors arosing oil content difference , builds the foundation for the later period research . Research results is as follows:Forward- and reward- cDNA libraries were constructed which "zhongdou 32" and "youchun02-6" DAF36 seed as tester by SSH . The subtraction efficiency of the forward and reverse subtracted libraries all above 2~5. The subtracted libraries were verified by PCR, the lengths of the inserted fragments were between 100bp and 900bp, average 434bp, The construction is successful.The 1337 clones were picked out by dot blot screening and PCR. Afer the combination and the classification to the sequence information by biological information, Total 32 genes were higher expressed in "zhongdou 32", and 30 genes in "youchun 02-6". Many genes were relative to the synthesis, degradation and transport of oil and protein, energy metabolism and reflection of defend and stress.Analysed the expression of 62 differentially expressed genes in soybean tissues, 6 differentially expressed genes were got which just appearance in soybean seed. These 6 genes respectively are Sucrose binding protein, p24 oleosin isoform B,Glycine max seed maturation protein PM34, beta-l,4-mannan synthase (ManS), lipoxygenase 1, GDSL-motif lipase. In addition, the differences of these genes were studied by real-time quantitative PCR which in two soybean lines and different seed developmental stages.