About 1 kb fragment of rbcS (ribulose 1,5 bisphosphate carboxylase small subunit) gene in wild soybean (Glycine soja, Ji 50017) was amplified from total DNA by PCR assay. Sequence analysis of the fragment indicated th...About 1 kb fragment of rbcS (ribulose 1,5 bisphosphate carboxylase small subunit) gene in wild soybean (Glycine soja, Ji 50017) was amplified from total DNA by PCR assay. Sequence analysis of the fragment indicated that 1089 bp sequenced included the whole coding region for Rubisco small subunit. The rbcS gene in wild soybean encoded a precursor composed of a transit peptide of 55 amino acids and a mature protein of 123 amino acids. There were two introns found in the rbcS gene as other dicotyledonous species previously sequenced.Comparison of DNA sequences showed high degree homology of rbcS genes between wild soybean and cultivated soybean (Glycine max var. wayne). Some changes of amino acids emerged from the diverse nucleotides did not affect the function of the small subunit. These results may contribute some basic data in molecular biology to study the origin and evolution of soybean.展开更多
为明确Rubisco酶小亚基(Rubisco small subunit,RbcS)在烟草抵抗主要病毒侵染胁迫中的作用,以本氏烟为研究材料,利用qRT-PCR、Western blot和瞬时表达体系对烟草RbcS基因在TMV、CMV和PVY侵染中的调控作用进行系统分析。结果表明,3种病...为明确Rubisco酶小亚基(Rubisco small subunit,RbcS)在烟草抵抗主要病毒侵染胁迫中的作用,以本氏烟为研究材料,利用qRT-PCR、Western blot和瞬时表达体系对烟草RbcS基因在TMV、CMV和PVY侵染中的调控作用进行系统分析。结果表明,3种病毒侵染本氏烟均引起NbRbcS在mRNA水平和蛋白水平上下调,瞬时过表达NbRbcS,病毒拷贝数均显著降低。而沉默NbRbcS,TMV、CMV和PVY拷贝数分别达到阴性对照的11.12、9.57和17.76倍。对PVY引起的斑驳叶片中深绿色和浅绿色区域分别进行qRT-PCR分析发现,在浅叶区NbRbcS mRNA的表达量只有深叶区的49.10%,但PVY的拷贝数却达到了深叶区的5.18倍。以上研究表明,RbcS大量表达可增强烟草对病毒的防御能力,烟草RbcS基因可作为潜在的抗病毒基因利用。展开更多
Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments,and consequently may have highly effi cient ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)activity for carbo...Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments,and consequently may have highly effi cient ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)activity for carbon assimilation.In our study,we cloned the full-length Rubisco gene from S.japonica(SJ-rbc).It contained an open reading frame for a large subunit gene(SJ-rbcL)of 1 467 bp,a small subunit gene(SJ-rbcS)of 420 bp,and a SJ-rbcL /S intergenic spacer of 269 bp.The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa,5.81 and 15.84 kDa,4.71,respectively.After induction with 1 mmol/L isopropyl-β-Dthiogalactopyranoside for 5 h and purifi cation by Ni 2+ affi nity chromatography,electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein.Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night.This observation implied that Rubisco played a role in normal gametophytic growth and development.In juvenile sporophytes,mRNA levels of SJ-rbcL,carbonic anhydrase,Calvin-BensonBassham cycle-related enzyme,and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance.Similarly,expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m 2·s).Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements.展开更多
文摘About 1 kb fragment of rbcS (ribulose 1,5 bisphosphate carboxylase small subunit) gene in wild soybean (Glycine soja, Ji 50017) was amplified from total DNA by PCR assay. Sequence analysis of the fragment indicated that 1089 bp sequenced included the whole coding region for Rubisco small subunit. The rbcS gene in wild soybean encoded a precursor composed of a transit peptide of 55 amino acids and a mature protein of 123 amino acids. There were two introns found in the rbcS gene as other dicotyledonous species previously sequenced.Comparison of DNA sequences showed high degree homology of rbcS genes between wild soybean and cultivated soybean (Glycine max var. wayne). Some changes of amino acids emerged from the diverse nucleotides did not affect the function of the small subunit. These results may contribute some basic data in molecular biology to study the origin and evolution of soybean.
文摘为明确Rubisco酶小亚基(Rubisco small subunit,RbcS)在烟草抵抗主要病毒侵染胁迫中的作用,以本氏烟为研究材料,利用qRT-PCR、Western blot和瞬时表达体系对烟草RbcS基因在TMV、CMV和PVY侵染中的调控作用进行系统分析。结果表明,3种病毒侵染本氏烟均引起NbRbcS在mRNA水平和蛋白水平上下调,瞬时过表达NbRbcS,病毒拷贝数均显著降低。而沉默NbRbcS,TMV、CMV和PVY拷贝数分别达到阴性对照的11.12、9.57和17.76倍。对PVY引起的斑驳叶片中深绿色和浅绿色区域分别进行qRT-PCR分析发现,在浅叶区NbRbcS mRNA的表达量只有深叶区的49.10%,但PVY的拷贝数却达到了深叶区的5.18倍。以上研究表明,RbcS大量表达可增强烟草对病毒的防御能力,烟草RbcS基因可作为潜在的抗病毒基因利用。
基金Supported by the Agriculture Science&Technology Achievements Transformation Fund(No.2011GB24910005)the Modern Agricultural-Industry Technology Research Project(No.200903030)+2 种基金the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A406)the Shandong Agriculture Breeding Engineering Biological Resources Innovation of Research Projectthe National"Twelfth Five-Year"Plan for Science&Technology Support(No.2013BAB01B01)
文摘Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments,and consequently may have highly effi cient ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)activity for carbon assimilation.In our study,we cloned the full-length Rubisco gene from S.japonica(SJ-rbc).It contained an open reading frame for a large subunit gene(SJ-rbcL)of 1 467 bp,a small subunit gene(SJ-rbcS)of 420 bp,and a SJ-rbcL /S intergenic spacer of 269 bp.The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa,5.81 and 15.84 kDa,4.71,respectively.After induction with 1 mmol/L isopropyl-β-Dthiogalactopyranoside for 5 h and purifi cation by Ni 2+ affi nity chromatography,electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein.Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night.This observation implied that Rubisco played a role in normal gametophytic growth and development.In juvenile sporophytes,mRNA levels of SJ-rbcL,carbonic anhydrase,Calvin-BensonBassham cycle-related enzyme,and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance.Similarly,expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m 2·s).Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements.
