Sweet Sorghum SSADH Gene Clone Identify And Salt Tolerance Sorghum Species Screening

Succinic semialdehyde dehydrogenase(SSADH) was a key enzyme of GABAshunt;GABA shunt was a metabolism bypass of TCA cycle.GABA shunt play a important role in the central neurotransmission system.In plant,this shunt associated with various biotic and abiotic stress responsses. SSADH play a key role with plant developing,whereas the relationship of SSADH and plant stress tolerance remain unclear.This study get the Sweet sorghum SSADH gene CDS sequence by clone, Prokaryotic Expression the SSADH protein and analyzed the enzyme enzymatic dynamics analysis;using the method of q RT-PCR detect the gene expression levels with difference Sweet sorghum species.Utilized agrobacterium-mediated transformation of sorghum callus and obtain overexpression transgenic plants.The main results as follows:1.c DNA sequence of SSADH gene was clones in Sweet sorghum.This sequence was 1584 bp,encoding 527 amino acids,and the molecular is 52.45KD;Gene structure and gene funcation analysis suggest the gene have Succinic semialdehyde dehydrogenase activity domain and a signal peptide of 24 amino acids.2. Construct a prokaryotic expression vector p ET-28a-SSADH(1386),obtain SSADH protein and pure this protein.Analyzed enzyme activity with crude protein and pure protein,the results display the relative enzyme activity of pure protein further enhance,the enzyme activity is 0.042 units/mg protein.when add AMP and ATP as the inhibitor of SSADH,the enzyme activity has been suppressing.3.Utilize the subcellar location confirm the SSADH gene of Sweet sorghum location at mitochondrial, this experiment results Coincide with the gene funcation.The q RT-PCR analysis 6 Sweet sorghum species young leaf SSADH expression levels,and find the expression levels not obvious.4. Agrobacterium-mediated transformation of sorghum callus and Screening with gradient Hygromycin of 3 cycles, obtain overexpression transgenic plants and culture in greenhouse;and identify those transgenic plants.5.Pass through 0m M、100m M、200m M Na Cl treatment 11 sorghum species seeds, statistical the seeding after 10 days and get the root weight、root length、seeding weight and seeding length. Screening and get one salt tolerance species Tais and one salt sensitive species 10-80.Above study results for us know the Sweet sorghum SSADH gene funcation and as the basic for Sweet sorghum breeding.