Saccharina japonica gametophytes can survive a long period under unfavorable environmental conditions,while they also delay in growth and/or reproduction.Although the reproduction in delayed gametophyte of S.japonica ...Saccharina japonica gametophytes can survive a long period under unfavorable environmental conditions,while they also delay in growth and/or reproduction.Although the reproduction in delayed gametophyte of S.japonica was known to be strongly influenced by light intensity,light quality,and photoperiod,no previous studies have evaluated their interactive effects on gametogenesis.To evaluate these effects,we used an orthogonal experiment to expose delayed gametophytes of S.japonica to different light intensities,light qualities,and photoperiods for 12 days.The results showed that changes in light intensity rather than light quality and photoperiod significantly affected the relative growth rates of the delayed gametophytes.Blue light had the greatest promotion on reproduction rate.The optimal light conditions in the early vegetative growth phase in gametogenesis induction for the delayed gametophytes were at 60–80μmol photons m^(−2) s^(−1) with daylength of 12 or 16 hours under white or blue light.When the delayed gametophytes were maintained in a constant light condition from delayed state to gametogenesis,the beneficial photoperiods for vegetative growth and reproductive rate were both 16L(16 hours of light):8D(8 hours of dark).However,when the delayed S.japonica gametophytes achieve the optimal growth state during the first 6 days and then they were cultured at different light conditions for the following 6 days,the reproduction rate increased as the daylength decreased and attained a peak value in group of 8L:16D photoperiod,indicating that photoperiod adjustment at the transition period is crucial in the gametogenesis induction process of delayed gametophyte of S.japonica.展开更多
Long non-coding RNAs(lncRNAs)regulate a variety of biological processes,including sexual reproduction and differentiation.Saccharina japonica,a commercially important brown alga in China,shows remarkable sexual dimorp...Long non-coding RNAs(lncRNAs)regulate a variety of biological processes,including sexual reproduction and differentiation.Saccharina japonica,a commercially important brown alga in China,shows remarkable sexual dimorphism in haploid gametophytes.The sex of Saccharina japonica gametophytes is determined by UV sexual system.However,no results have been reported on the lncRNAs involved in the sex-related gene regulation of S.japonica.This study identified a number of lncRNAs and assessed their expression levels in male and female gametophytes.Among them,a total of 405 lncRNAs and 211 mRNAs showed differential expressions.Furthermore,the functions of target genes of differentially expressed lncRNAs(DELs)differed from those of differentially expressed genes(DEGs),suggesting that lncRNA may interact with other functional proteins,in addition to DEGs,to involve sex regulation in S.japonica.There were 32 and 90 potential cis-regulatory and trans-regulatory interactions between DELsDEGs,respectively.Five of these lncRNAs(LNC_002974,LNC_021059,LNC_038466,LNC_051584,and LNC_027400)interacted with putative male sex determination region(SDR)genes,suggesting that they act as regulators in gametophytes'sex regulation potentially.Findings from this study contribute to our understanding of the roles of lncRNAs in sex differentiation and lay the foundation for functional studies of candidate lncRNAs in the future.展开更多
Saccharina is one of the most important cold-water living marine brown algal genera. In this study we ana-lyzed the transcriptome of S. japonica, which belongs to the 1 000 Plants （OneKP） Project, by using a next-ge...Saccharina is one of the most important cold-water living marine brown algal genera. In this study we ana-lyzed the transcriptome of S. japonica, which belongs to the 1 000 Plants （OneKP） Project, by using a next-generation high-throughput DNA sequencing technique. About 5.16 GB of raw data were generated, and 65 536 scaffolds with an average length of 454 bp were assembled with SOAP de novo assembly method. In total, 19 040 unigenes were identified by BLAST;25 734 scaffolds were clustered into 37 Gene ontology functional groups;6 760 scaffolds were classified into 25 COG categories, as well as 2 665 scaffolds that were assigned to 306 KEGG pathways. Majority of the unigenes exhibited more similarities to algae including brown algae and diatom than other cyanobacteria, marine diatom, and plant. Saccharina japonica has the outstanding capability to accumulate halogen such as Br and I via halogenation processes from seawater. We acquired 42 different vanadium-dependent haloperoxidases （vHPO） in S. japonica transcriptome data, including 5 segments of vanadium-dependent iodoperoxidase （vIPO） and 37 segments of vanadium-de-pendent bromoperoxidase （vBPO）. Complicated analyses of identified fulllength S. japonica vBPO1 and S. japonica vBPO2 revealed the importance of vBPO among species of brown algae and the strong relationship between marine algal vBPOs and vIPOs. This study will enhance our understanding of the biological charac-teristics and economic values of S. japonica species.展开更多
Ocean acidification and eutrophication are two important environmental stressors.They inevitably impact marine macroalgae,and hence the coastal ecosystem of China.Saccharina japonica,as the main culture species in Chi...Ocean acidification and eutrophication are two important environmental stressors.They inevitably impact marine macroalgae,and hence the coastal ecosystem of China.Saccharina japonica,as the main culture species in China,is suffering the harmful golden tide caused by Sargassum horneri.However,it remains unclear whether the detrimental effects of S.horneri on S.japonica cultivation become more severe in future acidified and eutrophic scenario.In this study,we respectively investigated the effects of pCO_(2)(400μatm and 1000μatm)and nutrients(non-enriched and enriched seawater)on the growth,photosynthesis,respiration,chlorophyll contents,and tissue nitrogen of S.japonica and S.horneri.Results indicated that enrichment of nutrients contributed S.horneri to utilize HCO_(3)^(−).The carbon acquisition pathway shifted from HCO_(3)^(−)to CO_(2) in S.japonica,while S.horneri re-mained using HCO_(3)^(−)regulated by nutrient enrichment.S.horneri exhibited better photosynthetic traits than S.japonica,with a higher level of net photosynthetic rate and chlorophyll contents at elevated pCO_(2) and enriched nutrients.Tissue nitrogen also accumulated richly in the thalli of S.horneri under higher pCO_(2) and nutrients.Significant enhancement in growth was only detected in S.horneri under synergistic stress.Together,S.horneri showed competitive dominance in current study.These findings suggest that increasing risk of golden tide in acidified and eutrophic ocean can most likely result in great damage to S.japonica cultivation.展开更多
Environmental changes,such as ocean acidification and eutrophication,have created threats to kelp mariculture.In this study,the growth,photosynthesis,respiration and nutrient composition of Saccharina japonica were ev...Environmental changes,such as ocean acidification and eutrophication,have created threats to kelp mariculture.In this study,the growth,photosynthesis,respiration and nutrient composition of Saccharina japonica were evaluated at different levels of pCO2(400 and 800μL L−1)and nutrients(nutrient-enriched and non-enriched seawater).Elevated pCO2 decreased the relative growth rate(RGR),net photosynthetic rate and contents of tissue carbon and tissue nitrogen under non-enriched nutrient conditions,but it had no significant effect on these parameters under nutrient-enriched conditions.The dark respiration rate was positively affected by elevated pCO2 regardless of the nutrient conditions.However,the C:N was unaffected by elevated pCO2 at both nutrient levels.These results implied that ocean acidification could reduce the production and nutrient contents in the tissues of S.japonica,which was associated with nutrient conditions.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
Saccharina japonica is an economically important cold water brown alga extensively cultivated in China. It is cultivated upside down under a floating rope net with its holdfast and meristematic area facing sunlight an...Saccharina japonica is an economically important cold water brown alga extensively cultivated in China. It is cultivated upside down under a floating rope net with its holdfast and meristematic area facing sunlight and UV irradiation and its blade tip toward dark, and other worse cultivation environmental factors also make S . japonic a face more stresses. In this study, S . japonica transcriptomes corresponding to its four developmental stages were analyzed. In total, 7 800 genes predicted in the genome were transcribed. We found that 1 208 of the 7 800 expressed and 2 697 annotated were virus associating genes. Of 778 diff erentially expressed genes (DEGs), 372 were annotatable and 209 were virus associating. Such portion of virus associating genes indicated that the S . japonica genome contained a large portion of active virus originating genes. It was found that the transcripts abundance associated with sugar biosynthesis was about 2.13 folds of all the expressed, indicating that the biosyntheses of structural and storage sugars were very important cellular processes. The total abundance of genes involved in the biosynthesis of alginate and laminarin were similar among all developmental stages, however, that of genes involved in the biosynthesis of mannitol increased about 2-folds from mushroom and adult stages to mature and aging stages. This trend explained our observation that the content of alginate was almost constant at diff erent development stages, while that of mannitol increased sharply. In addition, we found that a set of defense and cell recurring genes highly expressed and many of them expressed diff erentially among stages. On average, the sum abundance of the transcripts of these genes at four stages were 3.40-and 4.96-folds of all the annotated and all the expressed, respectively. This indicated that S . japonica sporophytes persistently respond possible pathogen and environment stresses. The findings are important for timing S . japonica harvest and amending the current cultivation mode.展开更多
IGS is abundant in polymorphism,which is widely used in the analysis of intraspecific genetic diversity and phylogenetic relationships among geographical populations.In this study,the 45S rDNA repeat unit of Saccharin...IGS is abundant in polymorphism,which is widely used in the analysis of intraspecific genetic diversity and phylogenetic relationships among geographical populations.In this study,the 45S rDNA repeat unit of Saccharina japonica was obtained for the first time by BAC clone sequencing.The total length of the 45S rDNA repeat unit of S.japonica was 8995 bp,including 5420 bp of 18S-5.8S-25S rDNA,and 3575 bp of IGS(Intergenic Spacer),with the GC content of 51.4%.The IGS was composed of a 465 bp of 3′-outer transcribed spacer(ETS),an 874 bp 5′-ETS,and a 2236 bp non-transcribed spacer(NTS),with the GC content of 50.1%.Fiber-FISH(fiber-fluorescence in situ hybridization)analysis of the distribution of 45S rDNA repeat units on the bacterial artificial chromosome illustrated that each fiber had at least five continuously moniliform hybridization signal points.This study provided a new candidate molecular marker for detecting intraspecific polymorphisms of S.japonica.In addition,the successful fiber-FISH analysis of the 45S rDNA on BAC molecule would contribute to the construction of the physical map and map-based cloning of this kelp.展开更多
The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that ...The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.展开更多
As a carbon-storage compound and osmoprotectant in brown algae,mannitol is synthesized and then accumulated at high levels in Saccharina japonica(Sja);however,the underlying control mechanisms have not been studied.Ou...As a carbon-storage compound and osmoprotectant in brown algae,mannitol is synthesized and then accumulated at high levels in Saccharina japonica(Sja);however,the underlying control mechanisms have not been studied.Our analysis of genomic and transcriptomic data from Sja shows that mannitol metabolism is a cyclic pathway composed of four distinct steps.A mannitol-1-phosphate dehydrogenase(M1 PDH2)and two mannitol-1-phosphatases(M1 Pase1 and MIPase2)work together or in combination to exhibit full enzymatic properties.Based on comprehensive transcriptomic data from different tissues,generations,and sexes as well as under different stress conditions,coupled with droplet digital PCR(dd PCR)and proteomic confirmation,we suggest that Sja M1 Pase1 plays a major role in mannitol biosynthesis and that the basic mannitol anabolism and the carbohydrate pool dynamics are responsible for carbon storage and anti-stress mechanism.Our proteomic data indicate that mannitol metabolism remains constant during diurnal cycle in Sja.In addition,we discover that mannitol-metabolism-associated(MMA)genes show differential expression between the multicellular filamentous(gametophyte)and large parenchymal thallus(sporophyte)generations and respond differentially to environmental stresses,such as hyposaline and hyperthermia conditions.Our results indicate that the ecophysiological significance of such differentially expressed genes may be attributable to the evolution of heteromorphic generations(filamentous and thallus)and environmental adaptation of Laminariales.展开更多
Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known...Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.展开更多
基金financially supported by the National Key R&D Program of China(Nos.2018YFD0900305,2018YFD 0901500)the China Agriculture Research System of MOF and MARA,Central Public-interest Scientific Institution Basal Research Fund CAFS(No.2020TD27)the‘Young Talent of Fishery Sciences’project from Laboratory for Marine Fisheries and Aquaculture(No.2018-MFS-T12).
文摘Saccharina japonica gametophytes can survive a long period under unfavorable environmental conditions,while they also delay in growth and/or reproduction.Although the reproduction in delayed gametophyte of S.japonica was known to be strongly influenced by light intensity,light quality,and photoperiod,no previous studies have evaluated their interactive effects on gametogenesis.To evaluate these effects,we used an orthogonal experiment to expose delayed gametophytes of S.japonica to different light intensities,light qualities,and photoperiods for 12 days.The results showed that changes in light intensity rather than light quality and photoperiod significantly affected the relative growth rates of the delayed gametophytes.Blue light had the greatest promotion on reproduction rate.The optimal light conditions in the early vegetative growth phase in gametogenesis induction for the delayed gametophytes were at 60–80μmol photons m^(−2) s^(−1) with daylength of 12 or 16 hours under white or blue light.When the delayed gametophytes were maintained in a constant light condition from delayed state to gametogenesis,the beneficial photoperiods for vegetative growth and reproductive rate were both 16L(16 hours of light):8D(8 hours of dark).However,when the delayed S.japonica gametophytes achieve the optimal growth state during the first 6 days and then they were cultured at different light conditions for the following 6 days,the reproduction rate increased as the daylength decreased and attained a peak value in group of 8L:16D photoperiod,indicating that photoperiod adjustment at the transition period is crucial in the gametogenesis induction process of delayed gametophyte of S.japonica.
基金supported by the National Natural Science Foundation of China(No.132072962)the Agricultural Industrial Technology System in Shandong Province(No.SDAIT-26)+3 种基金the‘First Class Fishery Discipline’Programme[(2020)3]in Shandong Provincethe Special Talent Programme‘Yishi Yiyi’in Shandong Province,Chinathe National Key R&D Program of China(No.2018YFD0901500)the Shandong Province Agriculture Seed Project(No.2021LZ GC004)。
文摘Long non-coding RNAs(lncRNAs)regulate a variety of biological processes,including sexual reproduction and differentiation.Saccharina japonica,a commercially important brown alga in China,shows remarkable sexual dimorphism in haploid gametophytes.The sex of Saccharina japonica gametophytes is determined by UV sexual system.However,no results have been reported on the lncRNAs involved in the sex-related gene regulation of S.japonica.This study identified a number of lncRNAs and assessed their expression levels in male and female gametophytes.Among them,a total of 405 lncRNAs and 211 mRNAs showed differential expressions.Furthermore,the functions of target genes of differentially expressed lncRNAs(DELs)differed from those of differentially expressed genes(DEGs),suggesting that lncRNA may interact with other functional proteins,in addition to DEGs,to involve sex regulation in S.japonica.There were 32 and 90 potential cis-regulatory and trans-regulatory interactions between DELsDEGs,respectively.Five of these lncRNAs(LNC_002974,LNC_021059,LNC_038466,LNC_051584,and LNC_027400)interacted with putative male sex determination region(SDR)genes,suggesting that they act as regulators in gametophytes'sex regulation potentially.Findings from this study contribute to our understanding of the roles of lncRNAs in sex differentiation and lay the foundation for functional studies of candidate lncRNAs in the future.
基金The National Natural Science Foundation of China under contract Nos 41206116,31140070 and 31271397Technology Project of Ocean and Fisheries of Guangdong Province under contract No.A201201E03+1 种基金the Fundamental Research Funds for the Central Universities under contract No.201262003the algal transcriptome sequencing was supported by OneKP Project(www.onekp.com)
文摘Saccharina is one of the most important cold-water living marine brown algal genera. In this study we ana-lyzed the transcriptome of S. japonica, which belongs to the 1 000 Plants （OneKP） Project, by using a next-generation high-throughput DNA sequencing technique. About 5.16 GB of raw data were generated, and 65 536 scaffolds with an average length of 454 bp were assembled with SOAP de novo assembly method. In total, 19 040 unigenes were identified by BLAST;25 734 scaffolds were clustered into 37 Gene ontology functional groups;6 760 scaffolds were classified into 25 COG categories, as well as 2 665 scaffolds that were assigned to 306 KEGG pathways. Majority of the unigenes exhibited more similarities to algae including brown algae and diatom than other cyanobacteria, marine diatom, and plant. Saccharina japonica has the outstanding capability to accumulate halogen such as Br and I via halogenation processes from seawater. We acquired 42 different vanadium-dependent haloperoxidases （vHPO） in S. japonica transcriptome data, including 5 segments of vanadium-dependent iodoperoxidase （vIPO） and 37 segments of vanadium-de-pendent bromoperoxidase （vBPO）. Complicated analyses of identified fulllength S. japonica vBPO1 and S. japonica vBPO2 revealed the importance of vBPO among species of brown algae and the strong relationship between marine algal vBPOs and vIPOs. This study will enhance our understanding of the biological charac-teristics and economic values of S. japonica species.
基金This work is funded by the Major Scientific and Technological Innovation Project of Shandong Provincial Key Research and Development Program(No.2019JZZY020708).
文摘Ocean acidification and eutrophication are two important environmental stressors.They inevitably impact marine macroalgae,and hence the coastal ecosystem of China.Saccharina japonica,as the main culture species in China,is suffering the harmful golden tide caused by Sargassum horneri.However,it remains unclear whether the detrimental effects of S.horneri on S.japonica cultivation become more severe in future acidified and eutrophic scenario.In this study,we respectively investigated the effects of pCO_(2)(400μatm and 1000μatm)and nutrients(non-enriched and enriched seawater)on the growth,photosynthesis,respiration,chlorophyll contents,and tissue nitrogen of S.japonica and S.horneri.Results indicated that enrichment of nutrients contributed S.horneri to utilize HCO_(3)^(−).The carbon acquisition pathway shifted from HCO_(3)^(−)to CO_(2) in S.japonica,while S.horneri re-mained using HCO_(3)^(−)regulated by nutrient enrichment.S.horneri exhibited better photosynthetic traits than S.japonica,with a higher level of net photosynthetic rate and chlorophyll contents at elevated pCO_(2) and enriched nutrients.Tissue nitrogen also accumulated richly in the thalli of S.horneri under higher pCO_(2) and nutrients.Significant enhancement in growth was only detected in S.horneri under synergistic stress.Together,S.horneri showed competitive dominance in current study.These findings suggest that increasing risk of golden tide in acidified and eutrophic ocean can most likely result in great damage to S.japonica cultivation.
基金Ocean University of China in 2018 and the Major Scientific and Technological Innovation Project of Shandong Provincial Key Research and Development Program(No.2019JZZY020708).
文摘Environmental changes,such as ocean acidification and eutrophication,have created threats to kelp mariculture.In this study,the growth,photosynthesis,respiration and nutrient composition of Saccharina japonica were evaluated at different levels of pCO2(400 and 800μL L−1)and nutrients(nutrient-enriched and non-enriched seawater).Elevated pCO2 decreased the relative growth rate(RGR),net photosynthetic rate and contents of tissue carbon and tissue nitrogen under non-enriched nutrient conditions,but it had no significant effect on these parameters under nutrient-enriched conditions.The dark respiration rate was positively affected by elevated pCO2 regardless of the nutrient conditions.However,the C:N was unaffected by elevated pCO2 at both nutrient levels.These results implied that ocean acidification could reduce the production and nutrient contents in the tissues of S.japonica,which was associated with nutrient conditions.
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.
基金Supported by the Fundamental Research Funds for the Central Universities(No.201762017)
文摘Saccharina japonica is an economically important cold water brown alga extensively cultivated in China. It is cultivated upside down under a floating rope net with its holdfast and meristematic area facing sunlight and UV irradiation and its blade tip toward dark, and other worse cultivation environmental factors also make S . japonic a face more stresses. In this study, S . japonica transcriptomes corresponding to its four developmental stages were analyzed. In total, 7 800 genes predicted in the genome were transcribed. We found that 1 208 of the 7 800 expressed and 2 697 annotated were virus associating genes. Of 778 diff erentially expressed genes (DEGs), 372 were annotatable and 209 were virus associating. Such portion of virus associating genes indicated that the S . japonica genome contained a large portion of active virus originating genes. It was found that the transcripts abundance associated with sugar biosynthesis was about 2.13 folds of all the expressed, indicating that the biosyntheses of structural and storage sugars were very important cellular processes. The total abundance of genes involved in the biosynthesis of alginate and laminarin were similar among all developmental stages, however, that of genes involved in the biosynthesis of mannitol increased about 2-folds from mushroom and adult stages to mature and aging stages. This trend explained our observation that the content of alginate was almost constant at diff erent development stages, while that of mannitol increased sharply. In addition, we found that a set of defense and cell recurring genes highly expressed and many of them expressed diff erentially among stages. On average, the sum abundance of the transcripts of these genes at four stages were 3.40-and 4.96-folds of all the annotated and all the expressed, respectively. This indicated that S . japonica sporophytes persistently respond possible pathogen and environment stresses. The findings are important for timing S . japonica harvest and amending the current cultivation mode.
基金supported by the National Natural Science Foundation of China(Grant No.41376136,32172963 to Z-G Z)the National Key R&D Program of China(Grant No.2018YFD0901500 to Y-H B)the World Class Discipline Project of Aquaculture(to Z-G Z).
文摘IGS is abundant in polymorphism,which is widely used in the analysis of intraspecific genetic diversity and phylogenetic relationships among geographical populations.In this study,the 45S rDNA repeat unit of Saccharina japonica was obtained for the first time by BAC clone sequencing.The total length of the 45S rDNA repeat unit of S.japonica was 8995 bp,including 5420 bp of 18S-5.8S-25S rDNA,and 3575 bp of IGS(Intergenic Spacer),with the GC content of 51.4%.The IGS was composed of a 465 bp of 3′-outer transcribed spacer(ETS),an 874 bp 5′-ETS,and a 2236 bp non-transcribed spacer(NTS),with the GC content of 50.1%.Fiber-FISH(fiber-fluorescence in situ hybridization)analysis of the distribution of 45S rDNA repeat units on the bacterial artificial chromosome illustrated that each fiber had at least five continuously moniliform hybridization signal points.This study provided a new candidate molecular marker for detecting intraspecific polymorphisms of S.japonica.In addition,the successful fiber-FISH analysis of the 45S rDNA on BAC molecule would contribute to the construction of the physical map and map-based cloning of this kelp.
基金This research was supported by Shanghai Universities Peak Discipline Project of Aquaculture and“Double First-Class Initiative”program for its First-Class Discipline of aquaculture.
文摘The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.
基金supported by the National Natural Science Foundation of China (Grant No. 41376143)the Leading Talents Program in Taishan Industry of Shandong Province+3 种基金the Leading Talents Program in Entrepreneurship and Innovation of Qingdaothe China-ASEAN Maritime Cooperation Fund “China-ASEAN Center for Joint Research and Promotion of Marine Aquaculture Technology”the China Agriculture Research System (Grant No. CARS-50)the Seed Industry Innovation and Industrialization Project of Fujian Province, China
文摘As a carbon-storage compound and osmoprotectant in brown algae,mannitol is synthesized and then accumulated at high levels in Saccharina japonica(Sja);however,the underlying control mechanisms have not been studied.Our analysis of genomic and transcriptomic data from Sja shows that mannitol metabolism is a cyclic pathway composed of four distinct steps.A mannitol-1-phosphate dehydrogenase(M1 PDH2)and two mannitol-1-phosphatases(M1 Pase1 and MIPase2)work together or in combination to exhibit full enzymatic properties.Based on comprehensive transcriptomic data from different tissues,generations,and sexes as well as under different stress conditions,coupled with droplet digital PCR(dd PCR)and proteomic confirmation,we suggest that Sja M1 Pase1 plays a major role in mannitol biosynthesis and that the basic mannitol anabolism and the carbohydrate pool dynamics are responsible for carbon storage and anti-stress mechanism.Our proteomic data indicate that mannitol metabolism remains constant during diurnal cycle in Sja.In addition,we discover that mannitol-metabolism-associated(MMA)genes show differential expression between the multicellular filamentous(gametophyte)and large parenchymal thallus(sporophyte)generations and respond differentially to environmental stresses,such as hyposaline and hyperthermia conditions.Our results indicate that the ecophysiological significance of such differentially expressed genes may be attributable to the evolution of heteromorphic generations(filamentous and thallus)and environmental adaptation of Laminariales.
基金This research was supported by the National Key R&D Program of China(Grant No.2018YFD0901500)the National Natural Science Foundation of China(Grant No.41376136)the Double First-Class Discipline of Fisheries Science of China.
文摘Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.
