[Objectives]To study the differences of growth rate,morphology,ultrastructure,pigment content and antioxidant enzyme activity of free-living conchocelis of cultivated type of Neopyropia yezoensis under different light...[Objectives]To study the differences of growth rate,morphology,ultrastructure,pigment content and antioxidant enzyme activity of free-living conchocelis of cultivated type of Neopyropia yezoensis under different light qualities(white,red,blue,and green light).[Methods]The study was carried out through light quality design and culture,growth rate determination,microstructure and ultrastructure observation,chlorophyll a content and carotenoid content determination,phycobiliprotein content determination,malondialdehyde(MDA)content determination,superoxide dismutase(SOD)activity determination.[Results]After 21 d of culture,the specific growth rate(SGR)and chlorophyll a content of free-living conchocelis of N.yezoensis were significantly increased by white light(WL),followed by red light(RL)and green light(GL),and they were the lowest under blue light(BL).Compared with the WL group,the BL group had the highest content of phycoerythrin(PE),and the RL and GL groups had the highest content of phycocyanin(PC).The algal body of WL group was normal black brown,and the cell wall was the thickest.In RL and GL groups,the algal bodies were green,and their diameters and cell wall thicknesses were similar to those in WL group.In BL group,the algal body was bright red,the diameter was the smallest,the cell wall was the thinnest,and the ultrastructure showed that the number of plastoglobulus on the thylakoid was the largest.After BL irradiation,the highest MDA content and the lowest SOD activity were observed.The results revealed that WL is the most beneficial to the growth of free-living conchocelis,followed by RL and GL,while BL has adverse effects.[Conclusions]This study explored the most suitable light quality conditions for the propagation of free-living conchocelis.It is expected to provide germplasm guarantee for the production and seedling of N.yezoensis.展开更多
The contents of 16 kinds of free amino acids （FAAs） in 7 Pyropia yezoensis cultivars were determined by pre-column derivatization high performance liquid chromatographyin early, middle and late harvest period.The re...The contents of 16 kinds of free amino acids （FAAs） in 7 Pyropia yezoensis cultivars were determined by pre-column derivatization high performance liquid chromatographyin early, middle and late harvest period.The results showed that the total FAA content ranged from 48.71 to 62.05 mg/g among the 7 test ma- terials. Among the 16 kinds of FAAs, Ala, Arg, Glu and Asp, four kinds of flavor amino acids, showed the highest contents, and their total content ranged from 36.05 to 53.96 mg/g, accounting for 69.11%-86.47% of the total FAA content. The total contents of flavor amino acids and FAAs all reached the peaks in all the test ma- terials, except the control cultivar, at middle harvest period. The essential amino acid content was trended to be increased in all the test materials with the extended harvest period, and it reached the peak at the late harvest period. Sutong showed the highest flavor amino acid content and FAAs content, followed by Y-H002, Suyan and Sulian, while the control cultivar showed the lowest contents.展开更多
Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of ex...Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of expressed proteins also varied amongst the samples. The fewest soluble proteins were present in filamentous sporophytes. There were more types and amounts of soluble protein in conchospores than in filamentous sporophytes, but fewer than in bulgy sporophytes. More types of protein were detected in filamentous sporophytes cultivated in high temperatures than in those growing in normal situations. The most types and amounts of protein were found in blade gametophytes in all samples. Blade gametophytes harvested last year and stored at -20 ℃ showed only minor differences in expression of proteins when compared with those harvested in different seasons.展开更多
[Objective] The aim was to explore the extraction technique and purification method of lectin from Porpya yezoensis,as well as to investigate the properties of lectin. [Method] The effects of four factors such as buff...[Objective] The aim was to explore the extraction technique and purification method of lectin from Porpya yezoensis,as well as to investigate the properties of lectin. [Method] The effects of four factors such as buffer system,solid-liquid ratio,temperature,and extracting time on the lectin extraction result from P. yezoensis were investigated by the use of specific activity as the indicator. Then the lectin was purified by DEAE-Sepharose FF chromatography. The hemagglutination activity was used as the indicator,and the properties of the lectin such as carbohydrate specificity,divalent cations dependence,temperature,acidic and alkaline stability and others were detected. [Result] The optimal extraction conditions were：extraction ageat Tris-HCl buffer （25 mmol/L,pH=7.5 ）,solid-liquid ratio of 1：5,extracting temperature of 40 ℃,and extracting time of 16 h. The activity of the lectin of P. yezoensis didn＇t change after 30 min of being heated at 50 ℃,which showed a certain extent of thermal stability,and the suitable pH value was 7-9. The lectin of P. yezoensis could combine with maltose,D-galactose,D-xylose and L-Arabinose,in which the binding force with the maltose was the strongest. The lectin activity was depended on the addition of Ca2＋ and Mg2＋. [Conclusion] The research had provided theoretical basis for the clarification of immune mechanism of P. yezoensis,as well as its high effective utilization.展开更多
The genomic resources of Porphyra yezoensis expressed sequence tags (ESTs) were utilized to identify simple sequence repeats (SSRs), or microsatellites. This method took the advantage of using ESTs and microsatellites...The genomic resources of Porphyra yezoensis expressed sequence tags (ESTs) were utilized to identify simple sequence repeats (SSRs), or microsatellites. This method took the advantage of using ESTs and microsatellites either for the establishment of gene identities or for the acquisition of high polymorphism. The microsatellites can be used as gene markers when microsatellites are tagged to genes. Revealed by bioinformatics analysis, 1 162 out of 21 954 ESTs contained microsatellites and cluster analysis indi- cated that 984 of these ESTs fell into 112 contigs, while the other 178 ESTs were singletons. A total of 290 unique SSR-containing genes were identified. The AAC SSRs were the most populous type of microsatellites. GC-rich microsatellites were predominant among all the microsatellites.展开更多
This study analyzes the community structure, the quantity changes of the algae and the effect of important environmental factors and estimates the total biomass of the attached green algae in the survey areas. The stu...This study analyzes the community structure, the quantity changes of the algae and the effect of important environmental factors and estimates the total biomass of the attached green algae in the survey areas. The study uses data from surveys of the attached green algae on the Porphyra yezoensis aquaculture rafts and data regarding the environmental factors from October 2010 to April 2011 in the Subei Shoal. The attached green algae on the rafts included Ulva prolifera, Capsosiphon groenlandicus, U. linza, U. intestinalis, U. clathrata, and U. cornpressa. The biomass changes of the attached green algae exhibited an inverted parabola： the biomass was the highest （14 898 t） in April, and was the second highest （2 034 t） in November; it was lowest in February （only 729 t） and increased sharply from March to April. The species diversity differed significantly among the seasons. In September and October, when the P. yezoensis aquaculture rafts were initially set up, the attached green algae had a high biodiversity, while from December to the next February, a variety of green algae species coexisted on the rafts, although the biomass was low, and from March to April, as the biomass increased sharply, the species diversity dropped to the minimum. During this time, C. groenlandicus was apparently dominant with the maximum biomass proportion up to 80%, while the U. prolifera proportion increased exponentially to 20% to 40%. The water temperature had a direct regulating effect on the biomass and the species succession of the attached green algae. The estimation of the community dynamics and the biomass of the green algae provided the evidence needed to track the origin of the large-scale green tide in the southern Yellow Sea.展开更多
Free living conchocelis of Porphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with 6 0Co γ rays of different doses (ranging from 100 Gy to 1000 Gy) to induce mutation. Most of the conchocelis maintained the...Free living conchocelis of Porphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with 6 0Co γ rays of different doses (ranging from 100 Gy to 1000 Gy) to induce mutation. Most of the conchocelis maintained the capability of penetrating into shells, growing and forming colonies in shells, but their vitality was seriously impaired by the irradiation of γ rays. A few conchocelis pigments were mutagenized directly into different color pigment mutants whose progeny conchospores and foliose thalli had the same colors. However, some irradiated conchocelis did not show the change in color at the conchocelis stage. The pigment mutation could be observed only after the conchospores of these conchocelis had germinated into young foliose thalli. Irradiation of low dose (100 Gy) promoted the growth of thallus and many with altered morphology were observed. Conchospores of the irradiated conchocelis attached to the culture nets were cultured in the sea, and growth of these progenies was observed and measured.展开更多
The taste and odor analysis of the first,second,fourth and sixth crops of laver(Porphyra yezoensis)was obtained by electronic tongue,taste-related compound,electronic nose and gas chromatography-ion mobility spectrome...The taste and odor analysis of the first,second,fourth and sixth crops of laver(Porphyra yezoensis)was obtained by electronic tongue,taste-related compound,electronic nose and gas chromatography-ion mobility spectrometer(GC-IMS).Through the detection of the electronic tongue,the pleasant tastes such as umami and richness are more intense in the early crops,which is consistent with the analysis results of free amino acids and flavored nucleotides.The electronic nose can clearly distinguish between different laver crops.GC-IMS separated and identified 36 volatile components from lavers,mainly including aldehydes,ketones,alcohols,esters,acids and aromatic substances.For the early harvested crops,nonanal,octanal,hexanal and benzaldehyde have a significant contribution to odor because of the high concentration and low threshold of these compounds.For the later harvested crops,the concentration of most ketones and some aldehydes increased.1-Octen-3-ol,malondialdehyde and heptanal constituted the main odor characteristics.展开更多
Pyropia yezoensis(formerly Porphyra yezoensis)is an economically important red alga that is cultured extensively in China.The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses....Pyropia yezoensis(formerly Porphyra yezoensis)is an economically important red alga that is cultured extensively in China.The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses.An incidence of red rot disease was found in a P.yezoensis farm from mid-November to mid-December 2015 at Lianyungang,Jiangsu Province,China.Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen,leading to red rot symptoms.The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox 1).In artifi cial infection experiments on the P.yezoensis blades,the P.chondricola isolate was able to cause the same characteristic histopathology seen in natural infections.P.chondricola grew well at a wide range of temperatures in the range 8-31℃,salinities at 0-45 and pH 5-9.In an orthogonal test used to determine the effects of environmental factors(temperature,salinity,and zoospore concentration)on infection,the data revealed that temperature was the most important factor to affect red rot disease development,with the optimal conditions for disease expansion being 20℃,35 salinity,and a zoospore concentration of 10^6 zoospores/mL.The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia,which will provide support to maintain the healthy development of the Pyropia industry in China.展开更多
Vegetative cells of Porphyra yezoensis are isolated with sea snail enzyme and cultured on the solidified agar medium. The results of experiments show that the isolated cells can survive,divide and regenerate well on t...Vegetative cells of Porphyra yezoensis are isolated with sea snail enzyme and cultured on the solidified agar medium. The results of experiments show that the isolated cells can survive,divide and regenerate well on the medium solidified with agar. The first division on the solid medium starts after 7 days' culture, 4 days later than the liquid culture. The survival rate of isolated cells is 71.3% on the solid medium, lower than the 86.2% of that in seawater.Thalli, thalloids,conchocelis, spermatangia and multicellular masses are developed on the solid/medium in the first month, slowly but normally. Spermatangia sacs disappear within 4 weeks. Without adding nutrient liquid onto the surface of solid medium or injecting seawater under the agar layer in order to keep moisture, the thalli and cell groups release monospores to form new thalli instead of enlarging their areas after 5 weeks' culturing. Some monospores regenerate new thalli. Other monospores lose their pigments and minimize their volume and divide quickly to form light pink calli. After 16 weeks, numerous calli can be seen on the solid medium and after 24 weeks' culturing, almost only calli and conchocelis can be seen. If the calli are immersed in seawater, the monospores are released and may develop into young thallus.展开更多
Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter...Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter. The receptors included the prooplasts, tissues and free-liv-ing conchocelis filaments of P. yezoensis. Several factors, for example, the voltage, capacitance and bi-valent cations, etc., were studied. Results show that these four methods are all efficient for gene transfor-mation in P. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4 ×10-5.GUS activity was detected 26 days after transfation by using PEG method.展开更多
To screen the stable expression genes related to the stress （strong light, dehydration and temperature shock） we applied Absolute real-time PCR technology to determine the transcription numbers of the selected test ...To screen the stable expression genes related to the stress （strong light, dehydration and temperature shock） we applied Absolute real-time PCR technology to determine the transcription numbers of the selected test genes in Porphyra yezoensis, which has been regarded as a potential model species responding the stress conditions in the intertidal. Absolute real-time PCR technology was applied to determine the transcription numbers of the selected test genes in Porphyra yezoensis, which has been regarded as a potential model species in stress responding. According to the results of photosynthesis parameters, we observed that Y（II） and Fv/Fm were significantly affected when stress was imposed on the thalli of Porphyra yezoensis, but underwent almost completely recovered under normal conditions, which were collected for the following experiments. Then three samples, which were treated with different grade stresses combined with salinity, irradiation and temperature, were collected. The transcription numbers of seven constitutive expression genes in above samples were determined after RNA extraction and cDNA synthesis. Finally, a general insight into the selection of internal control genes during stress response was obtained. We found that there were no obvious effects in terms of salinity stress （at salinity 90） on transcription of most genes used in the study. The 18S ribosomal RNA gene had the highest expression level, varying remarkably among different tested groups. RPS8 expression showed a high irregular variance between samples. GAPDH presented comparatively stable expression and could thus be selected as the internal control. EF-la showed stable expression during the series of multiple-stress tests. Our research provided available references for the selection of internal control genes for transcripts determination of P. yezoensis.展开更多
Pyropia yezoensis, an intertidal seaweed, experiences regular dehydration and rehydration with the tides. In this study, the responses of P. yezoensis to dehydration and rehydration under high and low CO2 concentratio...Pyropia yezoensis, an intertidal seaweed, experiences regular dehydration and rehydration with the tides. In this study, the responses of P. yezoensis to dehydration and rehydration under high and low CO2 concentrations （（600-700）×10^-6 and （40-80）×10^-6, named Group I and Group II respectively） were investigated. The thalli of Group I had a significantly higher effective photosystem II quantum yield than the thalli of Group II at 71% absolute water content （AWC）. There was little difference between thalli morphology, total Rubisco activity and total protein content at 100% and 71% AWC, which might be the basis for the normal performance of photosynthesis during moderate dehydration. A higher effective photosystem I quantum yield was observed in the thalli subjected to a low CO2 concentration during moderate dehydration, which might be caused by the enhancement of cyclic electron flow. These results suggested that P. yezoensis can directly utilize COz in ambient air during moderate dehydration.展开更多
In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chai...In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.展开更多
The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure lin...The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences.展开更多
Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodoph...Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were con-structed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5’- and 3’-flanking regions (Tub5’ and Tub3’) up- and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT?3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3’. These con-structs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5’-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when com-pared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.展开更多
Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50...Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50%) increased the purity(A564:A280) of phycoerythrin to 1.49, 3.92 fold of the raw extract(0.38) and the purity(A615:A280) of phycocyanin to 0.70, 3.33 fold of the raw extract(0.21). Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phycocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21% and 0.09% of dried P. yezoensis blade, respectively. The photodynamic cytotoxic experiment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80% and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54% and 32.54% of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer(phycoerythrin and phycocyanin) producer.展开更多
Fe2+ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe2+ requirement in nori had not been seen. Porphyra yezoensis cells were separat...Fe2+ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe2+ requirement in nori had not been seen. Porphyra yezoensis cells were separated enzymatically and cultured in a series of sterilized seawater media containing various concentra- tions of Fe2+. The growth development and cell were investigated in this work. Through this experiment, two biologically-meant concentration scales were found, one is low concentrations, 12.1–102.1 μg/L, 10–100 times than that in seawater, favoring the development of isolated cells of Porphyra and the other was high concentra- tions, more than 10mg/L inhibiting the cell growth, leading to the deformity and shrinkage of the cells. At the concentration of 50 mg/L, the cells stopped growing and died eventually.展开更多
Genetic variation of 11 lines of Porphyra yezoensis from the coastline of Kagoshima of Japan, Qingdao, Nantong, Putuo and Nanji Islands of China were studied by using amplified fragment length polymorphism (AFLP). 778...Genetic variation of 11 lines of Porphyra yezoensis from the coastline of Kagoshima of Japan, Qingdao, Nantong, Putuo and Nanji Islands of China were studied by using amplified fragment length polymorphism (AFLP). 778 bands were obtained with AFLP analysis of 16 primer combinations, among which 15 were unique, about 98.07% were polymorphic. The AFLP data showed that the closest genetic distance was 0 - 180 between two Kagoshima samples, and the farthest one was 0.397 between Kagoshima No. 1 and Nantong No - 9 line. The genetic distance showed that the variation was within the inner species scope. Neighbor-joining cluster and UPGMA cluster indicated that samples from Kagoshima and Qingdao were with high similarity and either with the samples of Nantong, Putuo and Nanji Islands. P. yezoensis in China shared high genetic diversity, and the genetic distance showed positive correlation with the geographic distance.展开更多
To explore ef fective physiological indexes to distinguish the resistant lines and susceptible lines of Pyt hium porphyrae,the causal agent of red rot disease of Pyropia yezoensis,and establish the disease-resistance ...To explore ef fective physiological indexes to distinguish the resistant lines and susceptible lines of Pyt hium porphyrae,the causal agent of red rot disease of Pyropia yezoensis,and establish the disease-resistance breeding strategy,we obtained and analyzed the candidate resistant and susceptible lines by population selection.Gametophytes of the candidate lines were cultured in seawater containing Pyt.porphyrae zoospores.Antioxidase activities,including superoxide dismutase(SOD),peroxidase(POD)and polyphenol oxidase(PPO),were measured and compared between the two lines before and after infection.In the resistant lines,SOD and POD activities increased and then decreased,but PPO activity rose with the prolongation of the infection time.The phenylalanine ammonia lyase(PAL)activities also increased and then decreased after infection,but it had significantly different expression in the two lines without pathogen attack.The synthesis rates ofβ-1,3-glucanase,and cell-wall degrading enzyme were different from each other between the two lines after infection of P yt.porphyrae.Comparison in the contents of malondialdehyde(MDA)and reactive oxygen species(ROS)in the two lines showed that,the two contents varied synchronously in response to the pathogen attack.Changes of these enzymes activities or contents demonstrated that Pyr.yezoensis could resist against the pathogen of P yt.porphyrae with the antioxidant defense capacity.In addition,β-1.3-glucanase content showed extremely significant dif ference between the two lines,and the PAL had consistent expression dif ference.Therefore,phenylalanine ammonia lyase(PAL)andβ-1,3-glucanase can be considered as an ef fective index to distinguish susceptible line and resistant line,with which the workload of the resistant breeding could be reduced in the future.展开更多
基金Supported by National Algae System(CARS-50)Modern Agricultural(Laver)Industrial Technology System of Jiangsu Province(JATS[2023]381)Research Project of Nantong City(MS22022065).
文摘[Objectives]To study the differences of growth rate,morphology,ultrastructure,pigment content and antioxidant enzyme activity of free-living conchocelis of cultivated type of Neopyropia yezoensis under different light qualities(white,red,blue,and green light).[Methods]The study was carried out through light quality design and culture,growth rate determination,microstructure and ultrastructure observation,chlorophyll a content and carotenoid content determination,phycobiliprotein content determination,malondialdehyde(MDA)content determination,superoxide dismutase(SOD)activity determination.[Results]After 21 d of culture,the specific growth rate(SGR)and chlorophyll a content of free-living conchocelis of N.yezoensis were significantly increased by white light(WL),followed by red light(RL)and green light(GL),and they were the lowest under blue light(BL).Compared with the WL group,the BL group had the highest content of phycoerythrin(PE),and the RL and GL groups had the highest content of phycocyanin(PC).The algal body of WL group was normal black brown,and the cell wall was the thickest.In RL and GL groups,the algal bodies were green,and their diameters and cell wall thicknesses were similar to those in WL group.In BL group,the algal body was bright red,the diameter was the smallest,the cell wall was the thinnest,and the ultrastructure showed that the number of plastoglobulus on the thylakoid was the largest.After BL irradiation,the highest MDA content and the lowest SOD activity were observed.The results revealed that WL is the most beneficial to the growth of free-living conchocelis,followed by RL and GL,while BL has adverse effects.[Conclusions]This study explored the most suitable light quality conditions for the propagation of free-living conchocelis.It is expected to provide germplasm guarantee for the production and seedling of N.yezoensis.
基金Supported by National High-tech R&D Program of China(2012AA10A406-5)National Natural Science Foundation of China(31272664)+2 种基金Natural Science Foundation of Jiangsu Province(BK20140439)Jiangsu Province Science and Technology Support Program,China(BE2012420)Independent Innovation Foundation of Agricultural Science and Technology of Jiangsu Province(CX(14)2081)~~
文摘The contents of 16 kinds of free amino acids （FAAs） in 7 Pyropia yezoensis cultivars were determined by pre-column derivatization high performance liquid chromatographyin early, middle and late harvest period.The results showed that the total FAA content ranged from 48.71 to 62.05 mg/g among the 7 test ma- terials. Among the 16 kinds of FAAs, Ala, Arg, Glu and Asp, four kinds of flavor amino acids, showed the highest contents, and their total content ranged from 36.05 to 53.96 mg/g, accounting for 69.11%-86.47% of the total FAA content. The total contents of flavor amino acids and FAAs all reached the peaks in all the test ma- terials, except the control cultivar, at middle harvest period. The essential amino acid content was trended to be increased in all the test materials with the extended harvest period, and it reached the peak at the late harvest period. Sutong showed the highest flavor amino acid content and FAAs content, followed by Y-H002, Suyan and Sulian, while the control cultivar showed the lowest contents.
基金supported by the National Natural Science Foundation of China (No. 40706050, 40706048 and 30700619)the National Science & Technology Pillar Program (No. 2006BAD01A13 and 2008BAC49B04)+2 种基金National special fund for transgenic project (No. 2009ZX08009-019B)Natural Science Foundation of Shandong Province (No. 2009ZRA02075)and Qingdao Municipal Science and Technology plan project (No. 09-2-5-8-hy)
文摘Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of expressed proteins also varied amongst the samples. The fewest soluble proteins were present in filamentous sporophytes. There were more types and amounts of soluble protein in conchospores than in filamentous sporophytes, but fewer than in bulgy sporophytes. More types of protein were detected in filamentous sporophytes cultivated in high temperatures than in those growing in normal situations. The most types and amounts of protein were found in blade gametophytes in all samples. Blade gametophytes harvested last year and stored at -20 ℃ showed only minor differences in expression of proteins when compared with those harvested in different seasons.
基金Supported by Natural Science Founding of Huaihai Institute of Technology (KX07034)~~
文摘[Objective] The aim was to explore the extraction technique and purification method of lectin from Porpya yezoensis,as well as to investigate the properties of lectin. [Method] The effects of four factors such as buffer system,solid-liquid ratio,temperature,and extracting time on the lectin extraction result from P. yezoensis were investigated by the use of specific activity as the indicator. Then the lectin was purified by DEAE-Sepharose FF chromatography. The hemagglutination activity was used as the indicator,and the properties of the lectin such as carbohydrate specificity,divalent cations dependence,temperature,acidic and alkaline stability and others were detected. [Result] The optimal extraction conditions were：extraction ageat Tris-HCl buffer （25 mmol/L,pH=7.5 ）,solid-liquid ratio of 1：5,extracting temperature of 40 ℃,and extracting time of 16 h. The activity of the lectin of P. yezoensis didn＇t change after 30 min of being heated at 50 ℃,which showed a certain extent of thermal stability,and the suitable pH value was 7-9. The lectin of P. yezoensis could combine with maltose,D-galactose,D-xylose and L-Arabinose,in which the binding force with the maltose was the strongest. The lectin activity was depended on the addition of Ca2＋ and Mg2＋. [Conclusion] The research had provided theoretical basis for the clarification of immune mechanism of P. yezoensis,as well as its high effective utilization.
文摘The genomic resources of Porphyra yezoensis expressed sequence tags (ESTs) were utilized to identify simple sequence repeats (SSRs), or microsatellites. This method took the advantage of using ESTs and microsatellites either for the establishment of gene identities or for the acquisition of high polymorphism. The microsatellites can be used as gene markers when microsatellites are tagged to genes. Revealed by bioinformatics analysis, 1 162 out of 21 954 ESTs contained microsatellites and cluster analysis indi- cated that 984 of these ESTs fell into 112 contigs, while the other 178 ESTs were singletons. A total of 290 unique SSR-containing genes were identified. The AAC SSRs were the most populous type of microsatellites. GC-rich microsatellites were predominant among all the microsatellites.
基金The National Basic Research Program (973 Program) of China under contract No.2010CB428703the National Natural Science Foundation of China under contract No.41306171+3 种基金the National Research Foundation-Shandong Province United Fund under contract No.U1406403the Qingdao Public Domain to Support Science and Technology Project under contract No.13–4–1–68–hythe Marine and Basic Research Funds of the First Institute of Oceanography,SOA under contract No.2015G09MOST International S&T Cooperation Program under contract No.2010DFA24340
文摘This study analyzes the community structure, the quantity changes of the algae and the effect of important environmental factors and estimates the total biomass of the attached green algae in the survey areas. The study uses data from surveys of the attached green algae on the Porphyra yezoensis aquaculture rafts and data regarding the environmental factors from October 2010 to April 2011 in the Subei Shoal. The attached green algae on the rafts included Ulva prolifera, Capsosiphon groenlandicus, U. linza, U. intestinalis, U. clathrata, and U. cornpressa. The biomass changes of the attached green algae exhibited an inverted parabola： the biomass was the highest （14 898 t） in April, and was the second highest （2 034 t） in November; it was lowest in February （only 729 t） and increased sharply from March to April. The species diversity differed significantly among the seasons. In September and October, when the P. yezoensis aquaculture rafts were initially set up, the attached green algae had a high biodiversity, while from December to the next February, a variety of green algae species coexisted on the rafts, although the biomass was low, and from March to April, as the biomass increased sharply, the species diversity dropped to the minimum. During this time, C. groenlandicus was apparently dominant with the maximum biomass proportion up to 80%, while the U. prolifera proportion increased exponentially to 20% to 40%. The water temperature had a direct regulating effect on the biomass and the species succession of the attached green algae. The estimation of the community dynamics and the biomass of the green algae provided the evidence needed to track the origin of the large-scale green tide in the southern Yellow Sea.
文摘Free living conchocelis of Porphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with 6 0Co γ rays of different doses (ranging from 100 Gy to 1000 Gy) to induce mutation. Most of the conchocelis maintained the capability of penetrating into shells, growing and forming colonies in shells, but their vitality was seriously impaired by the irradiation of γ rays. A few conchocelis pigments were mutagenized directly into different color pigment mutants whose progeny conchospores and foliose thalli had the same colors. However, some irradiated conchocelis did not show the change in color at the conchocelis stage. The pigment mutation could be observed only after the conchospores of these conchocelis had germinated into young foliose thalli. Irradiation of low dose (100 Gy) promoted the growth of thallus and many with altered morphology were observed. Conchospores of the irradiated conchocelis attached to the culture nets were cultured in the sea, and growth of these progenies was observed and measured.
基金supported by the National Key R&D Program of China(No.2018YFD0901004)the Special Fun-ds for the Technology System of Modern Agricultural Industry(No.CARS-50)the Central Publicinterest Scientific Institution Basal Research Fund,YSFRI,CAFS(No.20603022020013)。
文摘The taste and odor analysis of the first,second,fourth and sixth crops of laver(Porphyra yezoensis)was obtained by electronic tongue,taste-related compound,electronic nose and gas chromatography-ion mobility spectrometer(GC-IMS).Through the detection of the electronic tongue,the pleasant tastes such as umami and richness are more intense in the early crops,which is consistent with the analysis results of free amino acids and flavored nucleotides.The electronic nose can clearly distinguish between different laver crops.GC-IMS separated and identified 36 volatile components from lavers,mainly including aldehydes,ketones,alcohols,esters,acids and aromatic substances.For the early harvested crops,nonanal,octanal,hexanal and benzaldehyde have a significant contribution to odor because of the high concentration and low threshold of these compounds.For the later harvested crops,the concentration of most ketones and some aldehydes increased.1-Octen-3-ol,malondialdehyde and heptanal constituted the main odor characteristics.
基金Supported by the China Agriculture Research System(No.CARS-50)the National Natural Sciences Foundation of China(No.31372517)+2 种基金the Fundamental Research Funds for the Central Universities(No.201562018)the National Infrastructure of Fishery Germplasm Resources(No.2017DKA30470)the Project of Aoshan Scientific and Technological Innovation Program of Qingdao National Laboratory for Marine Science and Technology(No.2015ASKJ02)
文摘Pyropia yezoensis(formerly Porphyra yezoensis)is an economically important red alga that is cultured extensively in China.The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses.An incidence of red rot disease was found in a P.yezoensis farm from mid-November to mid-December 2015 at Lianyungang,Jiangsu Province,China.Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen,leading to red rot symptoms.The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox 1).In artifi cial infection experiments on the P.yezoensis blades,the P.chondricola isolate was able to cause the same characteristic histopathology seen in natural infections.P.chondricola grew well at a wide range of temperatures in the range 8-31℃,salinities at 0-45 and pH 5-9.In an orthogonal test used to determine the effects of environmental factors(temperature,salinity,and zoospore concentration)on infection,the data revealed that temperature was the most important factor to affect red rot disease development,with the optimal conditions for disease expansion being 20℃,35 salinity,and a zoospore concentration of 10^6 zoospores/mL.The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia,which will provide support to maintain the healthy development of the Pyropia industry in China.
基金This work was supported by a fund for doctorate candidate scientific research from the Ministry of Education (98042303)doctor start fund of Suzhou University (Q4114928)
文摘Vegetative cells of Porphyra yezoensis are isolated with sea snail enzyme and cultured on the solidified agar medium. The results of experiments show that the isolated cells can survive,divide and regenerate well on the medium solidified with agar. The first division on the solid medium starts after 7 days' culture, 4 days later than the liquid culture. The survival rate of isolated cells is 71.3% on the solid medium, lower than the 86.2% of that in seawater.Thalli, thalloids,conchocelis, spermatangia and multicellular masses are developed on the solid/medium in the first month, slowly but normally. Spermatangia sacs disappear within 4 weeks. Without adding nutrient liquid onto the surface of solid medium or injecting seawater under the agar layer in order to keep moisture, the thalli and cell groups release monospores to form new thalli instead of enlarging their areas after 5 weeks' culturing. Some monospores regenerate new thalli. Other monospores lose their pigments and minimize their volume and divide quickly to form light pink calli. After 16 weeks, numerous calli can be seen on the solid medium and after 24 weeks' culturing, almost only calli and conchocelis can be seen. If the calli are immersed in seawater, the monospores are released and may develop into young thallus.
文摘Electroporation, PEG, PEG plus electroporation and Biolistics methods were tested ingene transformation of P. yezoensis. The exogenous gus was from plasmid of pBI121 and pCAMBI-A1301, both contain the CaMV35S promoter. The receptors included the prooplasts, tissues and free-liv-ing conchocelis filaments of P. yezoensis. Several factors, for example, the voltage, capacitance and bi-valent cations, etc., were studied. Results show that these four methods are all efficient for gene transfor-mation in P. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4 ×10-5.GUS activity was detected 26 days after transfation by using PEG method.
基金Supported by the National Natural Science Foundation of China(Nos.41176134,30830015)the Prospective Joint Research Project of Jiangsu Province(No.BY2011188)+1 种基金the National Marine Public Welfare Research Project(Nos.201105008-2,201105023-7)the National Basic Research Program of China(973 Program)(No.2011CB411908)
文摘To screen the stable expression genes related to the stress （strong light, dehydration and temperature shock） we applied Absolute real-time PCR technology to determine the transcription numbers of the selected test genes in Porphyra yezoensis, which has been regarded as a potential model species responding the stress conditions in the intertidal. Absolute real-time PCR technology was applied to determine the transcription numbers of the selected test genes in Porphyra yezoensis, which has been regarded as a potential model species in stress responding. According to the results of photosynthesis parameters, we observed that Y（II） and Fv/Fm were significantly affected when stress was imposed on the thalli of Porphyra yezoensis, but underwent almost completely recovered under normal conditions, which were collected for the following experiments. Then three samples, which were treated with different grade stresses combined with salinity, irradiation and temperature, were collected. The transcription numbers of seven constitutive expression genes in above samples were determined after RNA extraction and cDNA synthesis. Finally, a general insight into the selection of internal control genes during stress response was obtained. We found that there were no obvious effects in terms of salinity stress （at salinity 90） on transcription of most genes used in the study. The 18S ribosomal RNA gene had the highest expression level, varying remarkably among different tested groups. RPS8 expression showed a high irregular variance between samples. GAPDH presented comparatively stable expression and could thus be selected as the internal control. EF-la showed stable expression during the series of multiple-stress tests. Our research provided available references for the selection of internal control genes for transcripts determination of P. yezoensis.
基金Supported by the Science and Technology Strategic Pilot Program of Chinese Academy of Sciences(No.XDA11020404)the National High Technology Research and Development Program of China(863 Program)(No.2012AA100806)+1 种基金the Tianjin Natural Science Foundation(No.12JCZDJC22200)the Project for Developing Marine Economy by Science and Technology in Tianjin(No.KX2010-0005)
文摘Pyropia yezoensis, an intertidal seaweed, experiences regular dehydration and rehydration with the tides. In this study, the responses of P. yezoensis to dehydration and rehydration under high and low CO2 concentrations （（600-700）×10^-6 and （40-80）×10^-6, named Group I and Group II respectively） were investigated. The thalli of Group I had a significantly higher effective photosystem II quantum yield than the thalli of Group II at 71% absolute water content （AWC）. There was little difference between thalli morphology, total Rubisco activity and total protein content at 100% and 71% AWC, which might be the basis for the normal performance of photosynthesis during moderate dehydration. A higher effective photosystem I quantum yield was observed in the thalli subjected to a low CO2 concentration during moderate dehydration, which might be caused by the enhancement of cyclic electron flow. These results suggested that P. yezoensis can directly utilize COz in ambient air during moderate dehydration.
基金supported by the 863 Project (Nos. 2002AA628120, 2004AA628090, 20060110A4013 and 2006AA10A413)
文摘In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.
文摘The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences.
文摘Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were con-structed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5’- and 3’-flanking regions (Tub5’ and Tub3’) up- and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT?3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3’. These con-structs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5’-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when com-pared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.
基金financially supported by the National Key Technology R&D Program (2012BAC07B03)Shanghai Universities First-class Disciplines Project,Discipline name: Marine Science and Shanghai Municipal Education Commission (Preponderant Subject Program #S30701)Key Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture, P. R. China, Shanghai Engineering Research Center of Aquaculture, Shanghai University Knowledge Service Platform, Shanghai Ocean University Aquatic Animal Breeding Center (ZF1206)
文摘Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50%) increased the purity(A564:A280) of phycoerythrin to 1.49, 3.92 fold of the raw extract(0.38) and the purity(A615:A280) of phycocyanin to 0.70, 3.33 fold of the raw extract(0.21). Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phycocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21% and 0.09% of dried P. yezoensis blade, respectively. The photodynamic cytotoxic experiment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80% and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54% and 32.54% of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer(phycoerythrin and phycocyanin) producer.
基金This work was supported by the NSFC (No. 40206019).
文摘Fe2+ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe2+ requirement in nori had not been seen. Porphyra yezoensis cells were separated enzymatically and cultured in a series of sterilized seawater media containing various concentra- tions of Fe2+. The growth development and cell were investigated in this work. Through this experiment, two biologically-meant concentration scales were found, one is low concentrations, 12.1–102.1 μg/L, 10–100 times than that in seawater, favoring the development of isolated cells of Porphyra and the other was high concentra- tions, more than 10mg/L inhibiting the cell growth, leading to the deformity and shrinkage of the cells. At the concentration of 50 mg/L, the cells stopped growing and died eventually.
基金National Natural Science Foundation of China under contract No. 49976030Key Ph D Foundation of Ningbo City under contract No. 02J20110-15 and No. 01N40100-59
文摘Genetic variation of 11 lines of Porphyra yezoensis from the coastline of Kagoshima of Japan, Qingdao, Nantong, Putuo and Nanji Islands of China were studied by using amplified fragment length polymorphism (AFLP). 778 bands were obtained with AFLP analysis of 16 primer combinations, among which 15 were unique, about 98.07% were polymorphic. The AFLP data showed that the closest genetic distance was 0 - 180 between two Kagoshima samples, and the farthest one was 0.397 between Kagoshima No. 1 and Nantong No - 9 line. The genetic distance showed that the variation was within the inner species scope. Neighbor-joining cluster and UPGMA cluster indicated that samples from Kagoshima and Qingdao were with high similarity and either with the samples of Nantong, Putuo and Nanji Islands. P. yezoensis in China shared high genetic diversity, and the genetic distance showed positive correlation with the geographic distance.
基金Supported by the National Natural Science Foundation of China(Nos.31372517,41006082)
文摘To explore ef fective physiological indexes to distinguish the resistant lines and susceptible lines of Pyt hium porphyrae,the causal agent of red rot disease of Pyropia yezoensis,and establish the disease-resistance breeding strategy,we obtained and analyzed the candidate resistant and susceptible lines by population selection.Gametophytes of the candidate lines were cultured in seawater containing Pyt.porphyrae zoospores.Antioxidase activities,including superoxide dismutase(SOD),peroxidase(POD)and polyphenol oxidase(PPO),were measured and compared between the two lines before and after infection.In the resistant lines,SOD and POD activities increased and then decreased,but PPO activity rose with the prolongation of the infection time.The phenylalanine ammonia lyase(PAL)activities also increased and then decreased after infection,but it had significantly different expression in the two lines without pathogen attack.The synthesis rates ofβ-1,3-glucanase,and cell-wall degrading enzyme were different from each other between the two lines after infection of P yt.porphyrae.Comparison in the contents of malondialdehyde(MDA)and reactive oxygen species(ROS)in the two lines showed that,the two contents varied synchronously in response to the pathogen attack.Changes of these enzymes activities or contents demonstrated that Pyr.yezoensis could resist against the pathogen of P yt.porphyrae with the antioxidant defense capacity.In addition,β-1.3-glucanase content showed extremely significant dif ference between the two lines,and the PAL had consistent expression dif ference.Therefore,phenylalanine ammonia lyase(PAL)andβ-1,3-glucanase can be considered as an ef fective index to distinguish susceptible line and resistant line,with which the workload of the resistant breeding could be reduced in the future.
