Formation of primary pit connection during conchocelis phase of Porphyra yezoensis （Bangiophyceae, Rhodophyta）

1 INTRODUCTION Pit connection is an important feature in Rhodophyta and significant in algal phylogeny. Various documents have illustrated the existence of pit-connection in red alga previously (Migita, 1967; Ramus 1969a, b; Bourne et al., 1970; Lee and F…     

Gamma-rays induction of mutation in conchocelis ofPorphyra yezoensis

Free-living conchocelis ofPorphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with⁶⁰Co-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.     

Effect of cytokinins and auxins on the growth of free-living conchocelis ofPorphyra yezoensis

IAA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis ofPorphyra yezoensis which showed that its growth in medium with cytokinins, IAA and NAA was more rapid than that in medium with non—phytohormones; that the optimal concentrations for promoting growth were 10 μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, IAA and cytokinins, NAA/ZA 1–1000/1 μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. IAA/ZA 0.1-1/0.1-1 μg/L; IAA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000 μg/L also promoted growth. Contribution No. 2335 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 94 from the Experimental Marine Biology Lab Qingdao, China. The research was supported by a grant from the Chinese Academy of Sciences.     

Studies on intercropping ofPorphyra haitanensis T. J. Chang et B. F. Zheng with Bay ScallopArgopecten irradians Lamarck

Results of the experiment of northward transplantation ofPorphyra haitanensis showed that intercrop ofP. haitanensis with Bay Scallop is feasible because the growth duration of both of them is about the same in August–December. Full use of cultivating area, lowering its cost and increasing its product per unit area as well as improving the ecological environment are realized during intercrop. Contribution No. 2152 from the Institute of Oceanology, Academia Sinica.     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta）

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.     

Subunit composition and chromophore content of R-phycoerythrin fromPorphyra haitanensis

R-phycoerythrin fromPorphyra haitanensis exists in two aggregation states with different molecular weights. A more highly aggregated form, RPE I, was chromatographed on Bio-Rex 70 column with urea solution (pH 3.0) as eluent, and the molecular weights of the 3 subunits (α, β, γ) obtained were determined on SDS-PAGE at 18000, 19200 and 30000, respectively. α subunit carried two phycoerythrobilin (PEB); β subunit, three PEB and one phycourobilin (PUB); γ subunit, one PEB and three PUB chromophores. The molar ratio of α, β, and γ subunits of RPE I was 6: 6: 1, and their subunit composition was confired to be (αβ)₆γ on account of the molecular weight of RPE I, 232000. A lower aggregated form, RPE II, contained α and β subunits similar to those of RPE I, but its subunit composition was the (αβ) monomer of RPE. Contribution No. 2108 from the Institute of Oceanology, Chinese Academy of Sciences     

Comparative studies on structural feature of agar polysaccharides fromPorphyra haitanensis grown in south and north China

The structural feature of agar polysaccharides fromPorphyra haitanensis grown in south China and transplanted to the north was investigated by fractionation on DEAE—Sephadex A 50, chemical analysis, and infrared and¹³C-NMR spectroscopy. The agars composed mainly of charged molecules were eluted from DEAE—Sephadex A 50 with 1.0 mol/L NaCl solution from the southernP. haitanensis and with 0.5 mol/L NaCl from the northern one. The¹³C-NMR spectra showed that agarobiose [(1→3)-β-D-galactopyranosyl-(1→4)-3,6-anhydro-α-L-galactopyranose] and the biological precursor of agarobiose [(1→3)-β-D-galactopyra nosyl-(1→4)-6-sulfate-α-L-galactopyranose] were the major disaccharide repeating units in the charged fractions. The content of the biological precursor in the agar polysaccharides from southernP. haitanensis was higher than that in the northern one, the content of the biological precursor extracted from cold water was higher than that from hot water, and the content of 6-OMe-D-galactose in the southernP. haitanensis polysaccharides was higher than in the northern one. This distinct difference will be of significance for further study of the physiological characters ofP. haitanensis. Contribution No. 1849 from the Institute of Oceanology, Academia Sinica.     

Study on cryopreservation of Porphyra yezoensis conchocelis

Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol （EG）, propylene glycol （PEG）, sorbitol and sucrose were developed. The effect of prefreezing at -40℃ or -20℃ for different time durations was compared and the thawing methods were screened. It was shown that the cryoprotectant including 10% DMSO with 0.5 molL^-1 sorbitol exhibited the optimal effect. The ideal pretreatment was that conchocelis segments were stayed for 20min at -40℃ before stored in liquid nitrogen, and 40℃ water bath was proper for quick thawing. The highest recovery rate of cryopreserved P. yezoensis conchocelis reached 89.41%.     

Use of rapd for detecting and identifyingPorphyra (Bangiales, Rhodophyta)

Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1−F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture. Contribution No. 100 from the Experimental Marine Biology Laboratory, IOCAS. Contribution No. 3289 from the Institute of Oceanology, Chinese Academy of Sciences. The research supported by the Foundation of the Chinese Academy of Sciences.     

Early stage differentiation of thallus cells ofPorphyra haitanensis (Rhodophyta)

The early stage differentiation of thallus cells ofPorphyra haitanensis T. J. Chang et B. F. Zheng was studied. Protoplasts or single cells were isolated from the blades using enzyme mixture comprising 2% sea snail gut enzyme and 1% cellulase. The isolated protoplasts or single cells were incubated in the MES medium. The cell differentiations were examined under the microscope at intervals after incubation. Four types of cell differentiation, namely, normal, abnormal, carposporangial and spermatorangial, and rhizoidal types, were observed. Since normal cell differentiations occur mostly in small thalli 50 mm in length and middle portions of big thalli 200 mm in length, it is essential to select tissues from these two kinds of thalli essential for commercial production.     

DEVELOPMENT OF SUSPENDED CONCHOCELIS OF PORPHYRA HAITANENSIS (BANGIALES, RHODOPHYTA)

INTRODUCTIONAferthelifehistoryofPorphrmwasdescrihed(Drew,l949;KUIDgi,1953;TsengandChang,l954),researchwasfocusedonthegrowthanddevelopmentofPorphpecon-chocelis(TsengandChang,1955;IwasakiandMatsudaira,l963;Waalandetal-,1987).PorphrmhaitanensisisaChinesespeciesveryimPOrtanforcultivationforcom-mercialproductioninsouthemChina.ConchocelisinshellsisthemainsoureeOfcon-chosPoresusedtoseedtheculturenets.However,ourknOWedgeofPoIPllyTahaitanensisisstilllithetedandtheinformationconcerningsee…     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda (Bangiales, Rhodophyta)

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensiss,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 exous 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.     

Studies on color type variants from mutagenized protoplasts ofPorphyra haitanensis Chang et Zheng &P. Yezoensis ueda (rhodophycease)

Isolated protoplasts from thalli ofPorphyra haitanensis andPorphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV). Several types of color variants were observed among the protoplast offspring. After treatment with colchicine: (1) 0.04–0.09% of red type variants inP. haitanensis were obtained; (2) The rate of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were 0.31–1.11% inP. yezoensis. After irradiation with UV: (1) 3.5–10.5% of red type variants inP. yezoensis were obtained: (2) 0.5–2.0% of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were obtained inP. haitanensis. Colchicine and UV’s mutangenic effects onP. yezoensis protoplasts were stronger than those onP. haitanensis protoplasts. The most efficient concentration of colchicine was 0.05%. The optimal length of UV-radiation was 1/2 min (radiation distance 5 cm). The red type variants induced by colchicine treatment grew faster than the wild type thalli. The clones of vegetative propagation from protoplasts of red type variants were still red type thalli. The red type variants will be good materials for genetic studies and improvement ofPorphyra strains.     

An improved PCR method for direct identification of Porphyra （Bangiales, Rhodophyta） using conchocelis based on a RUBISCO intergenic spacer

An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra(Bangiales,Rhodophyta),including Porphyra yezoensis(Jiangsu,China),P.haitanensis(Fujian,China),P.oligospermatangia(Qingdao,China),P.katadai(Qingdao,China),P.tenera(Qingdao,China),P.suborboculata(Fujian,China),P.pseudolinearis(Kogendo,Korea),P.linearis(Devon,England),and P.fallax(Seattle,USA)....     

Formation and early development of tetraspores of Polysiphonia urceolata （Rhodomelaceae, Rhodophyta）

Polysiphonia urceolata is one type of potential commercial red seaweeds used for breeding and cultivation, because of its significant biochemical and biomedical application. However, the information of breeding and seedling incubation for cultivation is limited, especially the early development. In this study, tetrasporohyte and gametophyte of P. urceolata were taken as the study materials in Huiquan Bay, Qingdao, China. The cleaned and sterilized tetrasporophytes and gametophytes were pre-cultured in sterilized seawater, then nurtured at 18°C, 25 μmol photons m⁻² s⁻¹ in 12:12 h (light:dark) photoperiod. Continuous observation under microscope showed that the early development consists of bipolar division stage and seedling stage. In the division stage, tetraspores germinate into bipolar sporelings that further differentiate into a colorless rhizoidal portion and a lightly pigmented upright shoot. The lightly pigmented rhizoidal cell develops to a rhizoid and the larger pigmented cell transforms to an erect axis. In the seedling stage, several quasi-protuberances appear on the erect axis and form juvenile seedlings. The results demonstrate the culture of P. urceolata from tetraspores under laboratory conditions. Supported by National Key Technology Support Program, Development Program of China (No.2006AA09Z21), National Natural Science Foundation of China (No. 40618001 and N_CUHK438/06) and Shandong Agricultural Seed Stock Breeding Project     

Axenic culture of free-living conchocelis of<Emphasis Type="Italic">Porphyra yezoensis</Emphasis> and<Emphasis Type="Italic">Porphyra haitanensis</Emphasis>

After discarding marine microorganisms from conchocelis of Porphyra yezoensis and Porphyra ha/tanens/s, their axenic cultures were obtained through treatment with antibiotics. Antibiotic disc tests were carried out to determine the effectiveness of each antibiotic in eliminating contaminating microorganisms. Five of 12 antibiotics tested were selected and used to produce the axenic cultures in this study, which showed that 200 μg/mL streptomycin, 250 μg/mL penicillin, 252 μg/mL kanamycin, 30 μg/mL neomycin, 200 μg/mL chloramphenicol were effective concentrations for eliminating microorganisms from conchocelis when antibiotics were added singly step by step; whereas simultaneous combination of 150 μg/mL streptomycin, 250 (or 350) μg/mL penicillin, 150 (or 250) μg/mL kanamycin, 70 μg/mL neomycin and 200 μg/mL chloramphenicol was also effective for producing the axenic cultures. However,it seemed that the treatments with antibiotics applied individually were more feasible than those with all antibiotics added at the same time. This may be due to the combined inhibiting effect of antibiotics on the growth and development of conchocelis.     

Hypoglossum fujianensis sp. nov. (Delesseriaceae, Rhodophyta) from Fujian Coast, China

Hypoglossum fujianensis sp. nov. is an epiphytic alga in the intertidal zone. Plants are light red, 0.9–2 cm high. Margin of branches gives rise to uniseriate hair-like rhizoids, formed outward from the fusion of the second-and third-order cells. Blades are single layered (except the midrib) and uncorticated. Tetrasporangial sori are formed on the middle part of blades. The globular tetrasporangia are developed from lateral pericentral cells. Project 39391800 supported by the NSFC.     

Three new species of Batrachospermum Roth （Batrachospermaceae, Rhodophyta） in China

Three new species of Batrachospermum Roth (Batrachospermales, Rhodophyta) from China are described in this paper. B. yunnanense sp. nov. has long-cylindrical trichogynes with long stalks and is diagnostic of section Virescentia. Within this section, B. yunnanense is similar to B. helminthosum Bory emend. Sheath et al., but it is dioecious and has curved carpogonial branches, while the latter is monoecious and has straight carpogonial branches. It is also similar to B. transtaganum Reis, but it differs from the latter in long carpogonia,big carposporophytes and carposporangia. It is considered that B. nothocladoideum sp. nov. is assigned to section Contorta, subsection Kushiroense, because its carpogonial branches are twisted and gonimoblast filaments are loosely agglomerated. This new species similar to B. iriomotense Kumano, but with short fascicles, long-ovoid or subpyriform cells, numerous terminal hairs, long-ellipsoid trichogynes, big carposporophytes and small carposporangia. The plant is quite tough and cartilaginous and similar to Nothocladus in gross morphology, but its carposporophytes are compact instead of diffuse. This shows that it may be a transitional species between section Contorta and genus Nothocladus. So, B. transitorium sp. nov. should belong to section Contorta, subsection Kushiroense, because of its curved or twisted carpogonial branches and loosely agglomerated gonimoblast filaments, with globose or subglobose cells in fascicles similar to B. spermatiophorum Vis et Sheath, but no colourless spermatiophores. In terms of small and numerous carposporophytes, B. transitorium sp. nov. is similar to some species of section Batrachospermum. However, their other features are unique, indicating its transitional nature between section Contorta and Batrachospermum.     

Effects of salinity, light and temperature on growth rates of two species of Gracilaria （Rhodophyta）

Effects of temperature, salinity and light intensity on growth rates of Gracilaria lichenoides and G. tenuistipitata var. liui Zhang et Xia were tested. Eight to ten levels of each factor were first tested separately. The best growth rate was obtained under the conditions of 32°C, 30 and 240 μmol/(m²·s) for G. lichenoides, and 24°C, 20 and 200 μmol/(m²·s) for G. tenuistipitata, respectively. Then a uniform design was used to evaluate the optimal combinations of the three factors. The best conditions for the highest daily specific growth rates (% increase in wet weight) are determined to be 31.30°C, 32.10, and 287.23 μmol/(m²·s) for G. lichenoides (16.26%/d), and 25.38°C, 21.10, and 229.07 μmol/(m²·s) for G. tenuistipitata (14.83%/d), respectively. Supported by the 908 Special Program (908-02-04-07), the National Basic Research Program of China (973 Program, No. 2006CB400608), and K. C. Wong Magna Fund in Ningbo University     

Effects of elevated CO2 on the photosynthesis and nitrate reductase activity of Pyropia haitanensis (Bangiales,Rhodophyta) grown at different nutrient levels

Pyropia haitanensis,a commercially important species,was cultured at two CO2 concentrations(390×10- 6 and 700×10-6(parts per million)) and at low and high nutrient levels,to explore the effect of elevated CO2 on the species under nutrient enrichment. Results show that in CO2-enriched thalli,relative growth rate(RGR) was enhanced under nutrient enrichment. Elevated CO2 decreased phycobiliprotein(PB) contents,but increased the contents of soluble carbohydrates. Nutrient enrichment increased the contents of chlorophyll a(Chl a) and PB,while soluble carbohydrate content decreased. CO2 enrichment enhanced the relative maximum electronic transport rate and light saturation point. In nutrient-enriched thalli the activity of nitrate reductase(NRA) increased under elevated CO2. An instantaneous pH change in seawater(from 8.1 to 9.6) resulted in reduction of NRA,and the thalli grown under both elevated CO2 and nutrient enrichment exhibited less pronounced reduction than in algae grown at the ambient CO2. The thermal optima of NRA under elevated CO2 and/or nutrient enrichment shifted to a lower temperature(10–15°C) compared to that in ambient conditions(20°C). We propose that accelerated photosynthesis could result in growth increment. Nassimilation remained high in acidified seawater and refl ected increased temperature sensitivity in response to elevated CO2 and eutrophication.