CAJ | 학술논문

大豆氧乙酰丝氨酸(硫醇)裂解酶(OAS-TL)催化半胱氨酸合成,是硫素代谢的关键酶.试验以东农46(高油型)和黑农35(高蛋白型)为材料,检测鼓粒期后大豆籽粒OAS-TL在3个不同硫水平(0、0.02、0.08 g·kg~(-1)土)下的生理特性,鉴定克隆了该酶基因并分析了其调控表达情况.结果表明,①OAS-TL活性随生育进程推进持续下降,2个品种籽粒OAS-TL活性表现为S_(20)高于S_0和S_(80)处理;鼓粒前期施硫效果大于后期;②通过同源克隆获得一个编码OAS-TL基因的cDNA序列,全长1 059 bp,ORF长855 bp,编码34.2 ku的蛋白质;③半定量RT-PCR显示OAS-TL稳定态mRNA在籽粒发育早期表达较强,且处理间差异明显,发育后期表达水平逐渐降低.不同硫素处理间OAS-TL的表达有差异,S_(20)上调表达高于S_0和S_(80).
대두양을선사안산(류순)렬해매(OAS-TL)최화반광안산합성,시류소대사적관건매.시험이동농46(고유형)화흑농35(고단백형)위재료,검측고립기후대두자립OAS-TL재3개불동류수평(0、0.02、0.08 g·kg~(-1)토)하적생리특성,감정극륭료해매기인병분석료기조공표체정황.결과표명,①OAS-TL활성수생육진정추진지속하강,2개품충자립OAS-TL활성표현위S_(20)고우S_0화S_(80)처리;고립전기시류효과대우후기;②통과동원극륭획득일개편마OAS-TL기인적cDNA서렬,전장1 059 bp,ORF장855 bp,편마34.2 ku적단백질;③반정량RT-PCR현시OAS-TL은정태mRNA재자립발육조기표체교강,차처리간차이명현,발육후기표체수평축점강저.불동류소처리간OAS-TL적표체유차이,S_(20)상조표체고우S_0화S_(80).
The objective of this study was to study and characterize O-acetyl-L-serine(thiol)-lyase (OAS-TL) which is a key enzyme in the pathway of sulfur metabolism that catalyzes the last step in the production of cysteine.Two cultivars, Dongnong46 (high-oil), Heinong35 (high-protein) were tested in this research by means of pots experiments with three sulfur application levels(0,0.02, 0.08 g·kg~(-1) soil) to study the effects of sulfur on the seeds of soybeans at the stage of drum particle. And than, cloned and characterized the expression OAS-TL. The results showed that:① The activity OAS-TL were higher at the early stage of seeds development and than declined along with the seeds maturity process. The treatment of S_(20) showed higher activity than those of S_0 and S_(80); The effect of each sulfur treatment showed more different at the early stage than the late stage;② A full-length cDNA clone encoding OAS-TL was obtained by homology cloning. The nucleotide sequenced covers 1 059 bp and nucleotides analysis of the cDNA revealed a single region of ORF of 855 bp encoding a 34.2 ku protein; ③RT-PCR analysis revealed that the OAS-TL mRNA was abundant and showed significant difference under each treatment at the early stage of seed development and than declined along with the seeds matudty process. The OAS-TL mRNA were different at each of the treatment, totally, the mRNA level of the treatment of S_(20) was more higher than those of S_0 and S_(80).