RAPD (random amplified polymorphic DNA) markers were generated from filaments of 15 Porphyra lines representing four important groups, P. yezoensis, P. haitanensis, P. katadai var. hemiphylla and P. olig...RAPD (random amplified polymorphic DNA) markers were generated from filaments of 15 Porphyra lines representing four important groups, P. yezoensis, P. haitanensis, P. katadai var. hemiphylla and P. oligospermatangia . Among the total 69 fragments generated by 6 selected primers (among 50 primers), 67 appeared to be polymorphic (97.1%). Cluster analysis based on the RAPD results was performed. The 15 Porphyra lines were divided into 3 groups. This result was consistent with that from taxonomy analysis. A DNA fingerprinting based on 8 bands amplified with OPN_02 and OPJ_18 was constructed and might be used in Porphyra variety identification. Five specific RAPD fragments of 5 Porphyra lines were isolated and cloned into pGEM_T easy vector. These five RAPD fragments may be useful in germplasm identification and property protection of Porphyra .展开更多
Vertical polyamide gel electrophoresis was used to investigate isozyme polymorphisms among different isolates （including wild and cultivated） of Porphyra katadai, Porphyra oligospermatangia, Porphyra yezoensis, Porp...Vertical polyamide gel electrophoresis was used to investigate isozyme polymorphisms among different isolates （including wild and cultivated） of Porphyra katadai, Porphyra oligospermatangia, Porphyra yezoensis, Porphyra haitanensis, and a hybridize species （Porphyra yezoensis x Porphyra haitanensis） sampled from China. Whereafter, the analyses of probable minimum loci numbers, observed alleles sum, genetic diversity, and unweighted pair-group arithmetic average （UPGMA） cluster were carded out. After initial activity and resolution testing of bands of 23 enzymes, 6 of them （MDH, ME, LDH, GDH, IDH and G-6-PDH） were proved to be appropriate for analysis of the full sample set. The probable minimum numbers of loci and alleles analyses showed that the five species of Porphyra had an extraordinary consistent result in ME loci and alleles. However, P. katadai and P. oligospermatangia differed from other three species of Porphyra in LDH and GDH loci and alleles. P. katadai was independent in the analyses of MDH and P. oligosperTnatangia and P. haitanensis differed from other three species in IDH analyses. Moreover, P. yezoensis and P. haitanensis were apart from other three species in G-6-PDH analysis. Taking one with another, P. katadai was relatively separated in the probable minimum numbers of loci and alleles analyses. The results indicated that the genetic variation among the five Porphyra species was limited with a genetic identity of 0.7550. The hybridize species （P. yezoensis x P. haitanensis） seemed to be high homologue with P. oligospermatangia, unexpectedly got relatively lower average genetic identities with both P. yezoensis and P. haitanensis. The 4 strains of P. yezoensis were relatively divergent with an average genetic identity of 0.7428, and P. katadai presented the most differentiated, compared with other species, which consistented with the result summarized in the probable minimum numbers of loci and alleles analyses.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
Several papers have reported that part or whole leafy thallus seemingly consisting of zygotospores can give rise to both blades and conchocelis in the same culture of Porphyra. Study on samples of wild and cul- tivate...Several papers have reported that part or whole leafy thallus seemingly consisting of zygotospores can give rise to both blades and conchocelis in the same culture of Porphyra. Study on samples of wild and cul- tivated Porphyra yezoensis and P. oligospermatangia were conducted to clarify the origination of the young blades in the culture. It is confirmed that single cells on the blade of both species, which normally intermixed with zygotospores, germinated into young blades. TEM and SEM observation has shown that the single cells of Porphyra yezoensis had typical features of female gamete (carpogonia) but archeospore. Therefore, the female gametes are responsible in developing leafy thalli.展开更多
Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV ). Several types of color variants were observed among the protoplast offsp...Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra 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 in P. 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 6.31- 1.11% in P. yezoensis. After irradiation with UV: (1) 3.5- 10.5% of red type variants in P. 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 in P. haitanensis. Colchicine and UV’s mutangenic effects on P. yezoensis protoplasts were stronger than those on P. 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
文摘RAPD (random amplified polymorphic DNA) markers were generated from filaments of 15 Porphyra lines representing four important groups, P. yezoensis, P. haitanensis, P. katadai var. hemiphylla and P. oligospermatangia . Among the total 69 fragments generated by 6 selected primers (among 50 primers), 67 appeared to be polymorphic (97.1%). Cluster analysis based on the RAPD results was performed. The 15 Porphyra lines were divided into 3 groups. This result was consistent with that from taxonomy analysis. A DNA fingerprinting based on 8 bands amplified with OPN_02 and OPJ_18 was constructed and might be used in Porphyra variety identification. Five specific RAPD fragments of 5 Porphyra lines were isolated and cloned into pGEM_T easy vector. These five RAPD fragments may be useful in germplasm identification and property protection of Porphyra .
基金sponsored jointly by the Natural Science Foundation of China (No. 40206019)Agricultural Innovation Project of Nantong (No. L4017)
文摘Vertical polyamide gel electrophoresis was used to investigate isozyme polymorphisms among different isolates （including wild and cultivated） of Porphyra katadai, Porphyra oligospermatangia, Porphyra yezoensis, Porphyra haitanensis, and a hybridize species （Porphyra yezoensis x Porphyra haitanensis） sampled from China. Whereafter, the analyses of probable minimum loci numbers, observed alleles sum, genetic diversity, and unweighted pair-group arithmetic average （UPGMA） cluster were carded out. After initial activity and resolution testing of bands of 23 enzymes, 6 of them （MDH, ME, LDH, GDH, IDH and G-6-PDH） were proved to be appropriate for analysis of the full sample set. The probable minimum numbers of loci and alleles analyses showed that the five species of Porphyra had an extraordinary consistent result in ME loci and alleles. However, P. katadai and P. oligospermatangia differed from other three species of Porphyra in LDH and GDH loci and alleles. P. katadai was independent in the analyses of MDH and P. oligosperTnatangia and P. haitanensis differed from other three species in IDH analyses. Moreover, P. yezoensis and P. haitanensis were apart from other three species in G-6-PDH analysis. Taking one with another, P. katadai was relatively separated in the probable minimum numbers of loci and alleles analyses. The results indicated that the genetic variation among the five Porphyra species was limited with a genetic identity of 0.7550. The hybridize species （P. yezoensis x P. haitanensis） seemed to be high homologue with P. oligospermatangia, unexpectedly got relatively lower average genetic identities with both P. yezoensis and P. haitanensis. The 4 strains of P. yezoensis were relatively divergent with an average genetic identity of 0.7428, and P. katadai presented the most differentiated, compared with other species, which consistented with the result summarized in the probable minimum numbers of loci and alleles analyses.
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.
基金This project was sponsored by the NSFC (No. C-0205-05-39770593)
文摘Several papers have reported that part or whole leafy thallus seemingly consisting of zygotospores can give rise to both blades and conchocelis in the same culture of Porphyra. Study on samples of wild and cul- tivated Porphyra yezoensis and P. oligospermatangia were conducted to clarify the origination of the young blades in the culture. It is confirmed that single cells on the blade of both species, which normally intermixed with zygotospores, germinated into young blades. TEM and SEM observation has shown that the single cells of Porphyra yezoensis had typical features of female gamete (carpogonia) but archeospore. Therefore, the female gametes are responsible in developing leafy thalli.
文摘Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra 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 in P. 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 6.31- 1.11% in P. yezoensis. After irradiation with UV: (1) 3.5- 10.5% of red type variants in P. 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 in P. haitanensis. Colchicine and UV’s mutangenic effects on P. yezoensis protoplasts were stronger than those on P. 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
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.
