The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte cl...The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte clone of L. japonica and a male gametophyte clone of L. longissima were hybridized, generating Dongfang No.2 hybrid Laminaria. The parentage of this hybrid Laminaria was determined using AFLP of total DNA, SNP of the ITS region of ribosomal RNA transcription unit and microsatellite DNA variation at two loci. In addition to 167 AFLP bands shared by Dongfang No.2, L. japonica and L. longissima, Dongfang No.2 hybrid Laminaria shared another 70 and 55 bands with L. japonica and L. longissima, respectively, which were obviously more than 11 bands shared by L. japonica and L. longissima. Dongfang No.2 held both ‘T’ and ‘C’ at position 847 of the ITS region, while ‘T’ at this position was specific for L. japonica and ‘C’ for L. longissima, respectively. Dongfang No.2 also held the microsatellite DNA alleles of two parents together at two microsatellite DNA marker loci. These observations clearly proved that Dongfang No.2 is a true hybrid of L. japonica and L. longissiuma. Unfortunately, the origin of the chloroplast of Dongfang No.2 was not determined based on the variation of RuBisCo spacer. More sequence variants of both ITS region and RuBisCo spacer were identified in Dongfang No.2 and most of them did not exist in either L. japonica or L. longissima. The unexpected variants may be due to the mutation of ga- metophyte clones occurring during their vegetative amplification.展开更多
We collected the diseased blades of Larninaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identifie...We collected the diseased blades of Larninaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identified as A lteromonas espejiana. This bacterium was applied as the causative pathogen to infect the blades of L. japonica under laboratory conditions. The aim of the present study was to identify the effects of the bacterium on the growth of L. japonica, and to find the possibly effective mechanism. Results showed that： （1）The blades of L.japonica exhibited symptoms of lesion,bleaching and deteriortion when infected by the bacterium. and their growth and photosynthesis were dramatically suppressed. At the same time, the reactive oxygen species （ROS） generation enhanced obviously, and the relative membrane permeability increased significantly. The contents of malonaldehyde （MDA） and free fatty acid in the microsomol membrane greatly elevated, but the phospholipid content decreased. Result suggested an obvious peroxidation and deesterrification in the blades of L. japonica when infected by the bacterium. （2） The simultaneous assay on the antioxidant enzyme activities demonstrated that superoxide dismutase （SOD） and catalase （CAT） increased greatly when infected by the bacterium, but glutathione peroxidase （Gpx） and ascorbate peroxidase （APX） did not exhibit active responses to the bacterium throughout the experiment. （3） The histomorphological observations gave a distinctive evidence of the severity of the lesions as well as the relative abundance in the bacterial population on the blades after infection. The bacterium firstly invaded into the endodermis of L. japonica and gathered around there, and then resulted in the membrane damage, cells corruption and ultimately, the death of L. japonica.展开更多
For the population genetics analysis of the naturally grown brown seaweed Laminaria japonica （Laminariales,Phaeophyta） sampled from Dalian,Yantai,Weihai,Rongcheng and Qingdao in China,ten primers were employed to pr...For the population genetics analysis of the naturally grown brown seaweed Laminaria japonica （Laminariales,Phaeophyta） sampled from Dalian,Yantai,Weihai,Rongcheng and Qingdao in China,ten primers were employed to produce 88 bands as revealed by randomly amplified polymorphic DNA （RAPD） markers,and all these bands were polymorphic.According to these band patterns,there were 94 distinct phenotypes occurred in 100 samples indicating the high heterozygosity of this kelp.Dalian population samples showed the highest percentage of polymorphism （71.67%）,and also the higher diversity estimated on the basis of the Shannon’s index （8.498）,suggesting that this population could be chosen as the best resource for genetic breeding.The highest diversity of Yantai population possibly resulted from the introduction of L.longissima used for interspecific cross breeding with L.japonica cultivated in China.From Dalian southwards to Qingdao,the genetic variation of the five populations became less with a decrease in latitude,possibly due to the natural selection especially of high temperature.The genetic distance （Φ ST values） of the five populations was a little significantly correlated with the geographical distance （r=0.496） at P =0.05 by Mantel’s test.Weihai,Rongcheng and Yantai populations were closely grouped genetically together by Neighbor-joining cluster analysis probably in that the dispersal of the kelp by propagules more easily occurring in the range of relatively short distance.The analysis of molecular variance （AMOVA） also demonstrated that the relatively higher variation occurred among populations （71.49%） at an extremely significant level （P 0.000 1）.All these evidence showed that there was a relatively distinct genetic differentiation among the sampled kelp populations,and L.japonica grown in China was also rather heterozygous in heredity.展开更多
Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for fina...Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.展开更多
Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-...Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molLl sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for 〉24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃ seawater and re-hydrated in 0.05 molL^-1 citrate sodium prepared using 30‰ NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.展开更多
The parthenogenesis and natural doubling of chromosomes in wild type female gametophytes of Laminaria japonica were studied. The results indicate that not all the female gametophytes from the wild type hybrid parent c...The parthenogenesis and natural doubling of chromosomes in wild type female gametophytes of Laminaria japonica were studied. The results indicate that not all the female gametophytes from the wild type hybrid parent can propagate through parthenogenesis. Most parthenosporophytes can mature, their spores germinate into gametophytes, the latter then developed into female sporophytes. To form these parthenosporophytes the natural doubling of chromosomes occurred mainly at the first and second cell divisions of the spores. It is thus considered that the parthenogenesis of L. japonica is inheritable and the relative genes link closely with the genes controlling the natural doubling of chromosomes and the female determination.展开更多
Laminaria （Saccharina） is one of the economic important seaweeds. Through sequence analysis of 4 099 ESTs from Laminaria （971 were generated from our own cDNA laboratories and 3 128 were downloaded from updated EST...Laminaria （Saccharina） is one of the economic important seaweeds. Through sequence analysis of 4 099 ESTs from Laminaria （971 were generated from our own cDNA laboratories and 3 128 were downloaded from updated EST databases） with SSRIT software, two hundred and fifty-four SSRs in 206 ESTs were found. From the 254 SSRs, sixty-three SSR primer-pairs were designed. In order to test their practicability, the 63 primer-pairs were tested in commonly used SSR reaction conditions using 12 Laminara DNA samples as templates. The results show that 23 SSR primer-pairs gave good amplification patterns on most （more than 80%） of the 12 Laminara DNA templates. Genetic diversity study of 12 Laminaria lines, which were widely used in breeding and economic cultivation in China, was performed based on the obtained SSR data.展开更多
This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time...This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time, the average length of filamentous female gametophytes was shortened.2. Ultrasonic treatment had emptying effect on cells. The number of empty cells increased with timeof treatment. Ultrasonic treatment had harmful effect on cells.3. Ultrasonic treatment could break down cell walls. The combination of frequency of 20 kHz, out-put of 15 W, 40 s and 60 s of treatment was best for this purpose. After ultrasonic treatment, the regner-ation of female gametophytes into sporophytes was effected. Female gametophytes could not recover aftertoo long period of treatment.展开更多
Possible mediatory roles of heat acclimation and salicylic acid in protecting the sporophyte of marine macroalga Laminaria japonica(Phaeophyceae) from heat stress were studied.Heat stress resulted in oxidative injury ...Possible mediatory roles of heat acclimation and salicylic acid in protecting the sporophyte of marine macroalga Laminaria japonica(Phaeophyceae) from heat stress were studied.Heat stress resulted in oxidative injury in the kelp blades.Under heat stress significant accumulation of hydrogen peroxide(H_2O_2) and malonaldehyde(MDA),a membrane lipid peroxidation product,and a drastic decrease in chlorophyll a content were recorded.Activity of the enzymatic antioxidant system was drastically affected by heat stress.The activity of superoxide dismutase(SOD) was significantly increased while peroxidase(POD),catalase(CAT) and glutathione peroxidase(GPX) were greatly inhibited and,simultaneously,phenylalanine ammonia-lyase was activated while polyphenol oxidase(PPO) was inhibited.Both heat acclimation pretreatment and exogenous application of salicylic acid alleviated oxidative damage in kelp blades.Blades receiving heat acclimation pretreatment and exogenous salicylic acid prior to heat stress exhibited a reduced increase in H_2O_2 and MDA content,and a lower reduction in chlorophyll a content.Pretreatment with heat acclimation and salicylic acid elevated activities of SOD,POD,CAT,GPX and PPO.Considering these results collectively,we speculate that the inhibition of antioxidant enzymes is a possible cause of the heat-stress-induced oxidative stress in L.japonica,and enhanced thermotolerance may be associated,at least in part,with the elevated activity of the enzymatic antioxidant system.展开更多
After continuous breeding gametophyte of Laminaria, then breaking protonema, heterosis seedling is got by bilineal hybridization. The result shows that the weight-increasing rate of female and male gametophyte clone c...After continuous breeding gametophyte of Laminaria, then breaking protonema, heterosis seedling is got by bilineal hybridization. The result shows that the weight-increasing rate of female and male gametophyte clone cells are 14.03% and 13.87%, after 16 d crossbreeding, the rate of ovulation is up to100%; the length of juvenile sporophyte after 40 d breeding is 2-3 cm. By farming on the sea, a hybrid combination is primarily screened.展开更多
Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Lamin...Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Laminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing inten- sity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the repro- ductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL1 (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that the Laminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development of Laminaria gametophytes.展开更多
In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and...In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and half months. In such a case, sometimes the sporelings died within a few days due to destructive diseases. In Order to overcome the mentioned problems, a new sporeling culture technique, the clone technique, is developed. The method includes three stead: (1) Gametophyte clone culture. The spores and the gametophytes are cloned in flasks under favorable environments. (2) Sporeling cul ture. Male and female clones are crushed and spread onto a frame to allow the gametophytes to attach to the substrata. The frames are cultured in tanks, and the sporophytes reach 1 cm in length within one and a half months. (3) Outgrowing of the plant. The frames are put in the open sea when seawater temperature decreased to 20℃. After one month, the sporelings are large enough to be transplanted. It is concluded that the clone technique has the following advantages: (1)Large amount of clones can be produced in a short period of time. (2) Clone seeding method makes it free from the biological rhythm, one can seed the plant anytime all the year round. (3) It takes only one and a half months to complete the process of sporeling cultivation in the greenhouse. At present, this technique is used in the breeding of new strains of Laminaria.展开更多
In this study, molecular weight controllable degradation of algal Laminaria japonica polysaccharides(LPS) was investigated by ultrasound combined with hydrogen peroxide. Three main factors, i.e., ultrasonic power(A), ...In this study, molecular weight controllable degradation of algal Laminaria japonica polysaccharides(LPS) was investigated by ultrasound combined with hydrogen peroxide. Three main factors, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C) were chosen for optimizing parameters by employing three-factors, three-levels BBD. The influence of degradation on structure change and antioxidant activities was also investigated. A second-order polynomial equation including molecular weight(Y) of Laminaria japonica polysaccharides and each variable parameter, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C), was established: Y=20718.67-4273.13A-4000.38B-1438.75C+2333.25AB+1511.00AC+873.00BC+2838.29A^2 + 2490.79B^2+873.04C^2. The equation regression coefficient value(R^2 = 0.969) indicated that this equation was valid. The value of the adjusted determination coefficient(adjusted R^2 = 0.914) also confirmed that the model was highly significant. The results of selected experimental degradation conditions matched with the predicted value. FT-IR spectra revealed that the structures of LPS before and after degradation were not significantly changed. Antioxidant activities of LPS revealed that low Mws possessed stronger inhibitory than the original polysaccharides. The scavenging effects on superoxide radicals was the highest when IC50 of crude LPS was 4.92 mg mL^(-1) and IC50 of Mw 18.576 KDa was 1.02 mg mL^(-1), which was fourfold higher than initial polysaccharide.展开更多
Effects of fucoidan from Laminaria japonica on 2,4 dinitrochlorobenzene induced delayed type hypersensitivity (DTH) reaction and the serum levels of IgG, IgM, complement C3 and C4 were investigated in the present stud...Effects of fucoidan from Laminaria japonica on 2,4 dinitrochlorobenzene induced delayed type hypersensitivity (DTH) reaction and the serum levels of IgG, IgM, complement C3 and C4 were investigated in the present study. Results showed that oral administration of fucoidan at dose of 150 and 300 mg/(kg·d) for 9 days before the hapten challenge significantly inhibited 2,4 dinitrochlorobenzene induced delayed type hypersensitivity reaction; and also inhibited the humoral immunity. Serum C3 and C4 levels were markedly reduced by fucoidan at dose of 150 and 300 mg/kg; and serum IgG and IgM levels were reduced by fucoidan at dose of 300 mg/kg. The inhibitory effects of fucoidan on delayed type hypersensitivity suggested that it may be potential medication for chronic inflammatory diseases in the future.展开更多
FGS was isolated and purified from laminaria japonica by enzymolysis, chromatography methods and so on. The high blood fat model was established by feeding mouse on high fat feedstuff. Compared with control group, the...FGS was isolated and purified from laminaria japonica by enzymolysis, chromatography methods and so on. The high blood fat model was established by feeding mouse on high fat feedstuff. Compared with control group, the concentration of HDL-C, the ratio of HDL-C/TC increased;while the concentration of TC, LDL-C and TG decreased in experimental groups. Conclusion: FGS was an effective serum lipid regulator and can be developed as medicine and health food.展开更多
The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments w...The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments was determined spectroscopically. The results show that: (1) among the six organic solvents, acetone was the most effective one for the extraction; (2) the optimum extraction conditions were as follows: the ratio of S/M (solvent volume/material weight) was 30 ml/g; fresh seaweed was extracted 2 times in 2h; (3) the average total content of pigments was 1.85 mg/g (calculated with dry L. japonica).展开更多
文摘The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte clone of L. japonica and a male gametophyte clone of L. longissima were hybridized, generating Dongfang No.2 hybrid Laminaria. The parentage of this hybrid Laminaria was determined using AFLP of total DNA, SNP of the ITS region of ribosomal RNA transcription unit and microsatellite DNA variation at two loci. In addition to 167 AFLP bands shared by Dongfang No.2, L. japonica and L. longissima, Dongfang No.2 hybrid Laminaria shared another 70 and 55 bands with L. japonica and L. longissima, respectively, which were obviously more than 11 bands shared by L. japonica and L. longissima. Dongfang No.2 held both ‘T’ and ‘C’ at position 847 of the ITS region, while ‘T’ at this position was specific for L. japonica and ‘C’ for L. longissima, respectively. Dongfang No.2 also held the microsatellite DNA alleles of two parents together at two microsatellite DNA marker loci. These observations clearly proved that Dongfang No.2 is a true hybrid of L. japonica and L. longissiuma. Unfortunately, the origin of the chloroplast of Dongfang No.2 was not determined based on the variation of RuBisCo spacer. More sequence variants of both ITS region and RuBisCo spacer were identified in Dongfang No.2 and most of them did not exist in either L. japonica or L. longissima. The unexpected variants may be due to the mutation of ga- metophyte clones occurring during their vegetative amplification.
基金The Hi-Tech Research and Development Program (863) of China (Youth Program) (No. 2004AA639770) and the Chinese NationalBasic Research Priorities Program (No. G1999012004)
文摘We collected the diseased blades of Larninaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identified as A lteromonas espejiana. This bacterium was applied as the causative pathogen to infect the blades of L. japonica under laboratory conditions. The aim of the present study was to identify the effects of the bacterium on the growth of L. japonica, and to find the possibly effective mechanism. Results showed that： （1）The blades of L.japonica exhibited symptoms of lesion,bleaching and deteriortion when infected by the bacterium. and their growth and photosynthesis were dramatically suppressed. At the same time, the reactive oxygen species （ROS） generation enhanced obviously, and the relative membrane permeability increased significantly. The contents of malonaldehyde （MDA） and free fatty acid in the microsomol membrane greatly elevated, but the phospholipid content decreased. Result suggested an obvious peroxidation and deesterrification in the blades of L. japonica when infected by the bacterium. （2） The simultaneous assay on the antioxidant enzyme activities demonstrated that superoxide dismutase （SOD） and catalase （CAT） increased greatly when infected by the bacterium, but glutathione peroxidase （Gpx） and ascorbate peroxidase （APX） did not exhibit active responses to the bacterium throughout the experiment. （3） The histomorphological observations gave a distinctive evidence of the severity of the lesions as well as the relative abundance in the bacterial population on the blades after infection. The bacterium firstly invaded into the endodermis of L. japonica and gathered around there, and then resulted in the membrane damage, cells corruption and ultimately, the death of L. japonica.
基金The National Natural Science Foundation of China under grant Nos 39800105 and 30471328the Shanghai Development Foundation of Education under grant No. 98-SG-32Key Discipline of Shanghai Municipal Education Commission (J50701)
文摘For the population genetics analysis of the naturally grown brown seaweed Laminaria japonica （Laminariales,Phaeophyta） sampled from Dalian,Yantai,Weihai,Rongcheng and Qingdao in China,ten primers were employed to produce 88 bands as revealed by randomly amplified polymorphic DNA （RAPD） markers,and all these bands were polymorphic.According to these band patterns,there were 94 distinct phenotypes occurred in 100 samples indicating the high heterozygosity of this kelp.Dalian population samples showed the highest percentage of polymorphism （71.67%）,and also the higher diversity estimated on the basis of the Shannon’s index （8.498）,suggesting that this population could be chosen as the best resource for genetic breeding.The highest diversity of Yantai population possibly resulted from the introduction of L.longissima used for interspecific cross breeding with L.japonica cultivated in China.From Dalian southwards to Qingdao,the genetic variation of the five populations became less with a decrease in latitude,possibly due to the natural selection especially of high temperature.The genetic distance （Φ ST values） of the five populations was a little significantly correlated with the geographical distance （r=0.496） at P =0.05 by Mantel’s test.Weihai,Rongcheng and Yantai populations were closely grouped genetically together by Neighbor-joining cluster analysis probably in that the dispersal of the kelp by propagules more easily occurring in the range of relatively short distance.The analysis of molecular variance （AMOVA） also demonstrated that the relatively higher variation occurred among populations （71.49%） at an extremely significant level （P 0.000 1）.All these evidence showed that there was a relatively distinct genetic differentiation among the sampled kelp populations,and L.japonica grown in China was also rather heterozygous in heredity.
文摘Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.
文摘Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molLl sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for 〉24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃ seawater and re-hydrated in 0.05 molL^-1 citrate sodium prepared using 30‰ NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.
文摘The parthenogenesis and natural doubling of chromosomes in wild type female gametophytes of Laminaria japonica were studied. The results indicate that not all the female gametophytes from the wild type hybrid parent can propagate through parthenogenesis. Most parthenosporophytes can mature, their spores germinate into gametophytes, the latter then developed into female sporophytes. To form these parthenosporophytes the natural doubling of chromosomes occurred mainly at the first and second cell divisions of the spores. It is thus considered that the parthenogenesis of L. japonica is inheritable and the relative genes link closely with the genes controlling the natural doubling of chromosomes and the female determination.
基金The National Natural Science Foundation of China under contract No.40776072 and 40606035the National High Technology Research and Development Program(863 Program)of China under contract Nos 2006AA10A412 and 2010AA10A401the Key Projects in the National Science & Technology Pillar Program under contract No.2006BAD01A13
文摘Laminaria （Saccharina） is one of the economic important seaweeds. Through sequence analysis of 4 099 ESTs from Laminaria （971 were generated from our own cDNA laboratories and 3 128 were downloaded from updated EST databases） with SSRIT software, two hundred and fifty-four SSRs in 206 ESTs were found. From the 254 SSRs, sixty-three SSR primer-pairs were designed. In order to test their practicability, the 63 primer-pairs were tested in commonly used SSR reaction conditions using 12 Laminara DNA samples as templates. The results show that 23 SSR primer-pairs gave good amplification patterns on most （more than 80%） of the 12 Laminara DNA templates. Genetic diversity study of 12 Laminaria lines, which were widely used in breeding and economic cultivation in China, was performed based on the obtained SSR data.
文摘This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time, the average length of filamentous female gametophytes was shortened.2. Ultrasonic treatment had emptying effect on cells. The number of empty cells increased with timeof treatment. Ultrasonic treatment had harmful effect on cells.3. Ultrasonic treatment could break down cell walls. The combination of frequency of 20 kHz, out-put of 15 W, 40 s and 60 s of treatment was best for this purpose. After ultrasonic treatment, the regner-ation of female gametophytes into sporophytes was effected. Female gametophytes could not recover aftertoo long period of treatment.
基金Supported by the Open Funds of the Key Oceanic Science and Engineering Laboratory of National Bureau of Oceanography(MESE-2008-01)the Specialized Research Program for Marine Public Welfare Industry,SOA(No.200805066)
文摘Possible mediatory roles of heat acclimation and salicylic acid in protecting the sporophyte of marine macroalga Laminaria japonica(Phaeophyceae) from heat stress were studied.Heat stress resulted in oxidative injury in the kelp blades.Under heat stress significant accumulation of hydrogen peroxide(H_2O_2) and malonaldehyde(MDA),a membrane lipid peroxidation product,and a drastic decrease in chlorophyll a content were recorded.Activity of the enzymatic antioxidant system was drastically affected by heat stress.The activity of superoxide dismutase(SOD) was significantly increased while peroxidase(POD),catalase(CAT) and glutathione peroxidase(GPX) were greatly inhibited and,simultaneously,phenylalanine ammonia-lyase was activated while polyphenol oxidase(PPO) was inhibited.Both heat acclimation pretreatment and exogenous application of salicylic acid alleviated oxidative damage in kelp blades.Blades receiving heat acclimation pretreatment and exogenous salicylic acid prior to heat stress exhibited a reduced increase in H_2O_2 and MDA content,and a lower reduction in chlorophyll a content.Pretreatment with heat acclimation and salicylic acid elevated activities of SOD,POD,CAT,GPX and PPO.Considering these results collectively,we speculate that the inhibition of antioxidant enzymes is a possible cause of the heat-stress-induced oxidative stress in L.japonica,and enhanced thermotolerance may be associated,at least in part,with the elevated activity of the enzymatic antioxidant system.
文摘After continuous breeding gametophyte of Laminaria, then breaking protonema, heterosis seedling is got by bilineal hybridization. The result shows that the weight-increasing rate of female and male gametophyte clone cells are 14.03% and 13.87%, after 16 d crossbreeding, the rate of ovulation is up to100%; the length of juvenile sporophyte after 40 d breeding is 2-3 cm. By farming on the sea, a hybrid combination is primarily screened.
基金This research was supported by the NSFC (No.40376049) and Japan Society for the Promotion of Science (The Invitation Fellowship for Research in Japan 2002)
文摘Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Laminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing inten- sity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the repro- ductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL1 (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that the Laminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development of Laminaria gametophytes.
基金Key Project, the Ninth Five-Year Plan of China! 96-008-01-02-06Project of Bio-Engineering Center of China! SSTC 96-C01-05-01
文摘In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and half months. In such a case, sometimes the sporelings died within a few days due to destructive diseases. In Order to overcome the mentioned problems, a new sporeling culture technique, the clone technique, is developed. The method includes three stead: (1) Gametophyte clone culture. The spores and the gametophytes are cloned in flasks under favorable environments. (2) Sporeling cul ture. Male and female clones are crushed and spread onto a frame to allow the gametophytes to attach to the substrata. The frames are cultured in tanks, and the sporophytes reach 1 cm in length within one and a half months. (3) Outgrowing of the plant. The frames are put in the open sea when seawater temperature decreased to 20℃. After one month, the sporelings are large enough to be transplanted. It is concluded that the clone technique has the following advantages: (1)Large amount of clones can be produced in a short period of time. (2) Clone seeding method makes it free from the biological rhythm, one can seed the plant anytime all the year round. (3) It takes only one and a half months to complete the process of sporeling cultivation in the greenhouse. At present, this technique is used in the breeding of new strains of Laminaria.
基金the financial support from the National Natural Science Foundation of China (No.21506220)
文摘In this study, molecular weight controllable degradation of algal Laminaria japonica polysaccharides(LPS) was investigated by ultrasound combined with hydrogen peroxide. Three main factors, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C) were chosen for optimizing parameters by employing three-factors, three-levels BBD. The influence of degradation on structure change and antioxidant activities was also investigated. A second-order polynomial equation including molecular weight(Y) of Laminaria japonica polysaccharides and each variable parameter, i.e., ultrasonic power(A), ultrasonic time(B), and H_2O_2 concentration(C), was established: Y=20718.67-4273.13A-4000.38B-1438.75C+2333.25AB+1511.00AC+873.00BC+2838.29A^2 + 2490.79B^2+873.04C^2. The equation regression coefficient value(R^2 = 0.969) indicated that this equation was valid. The value of the adjusted determination coefficient(adjusted R^2 = 0.914) also confirmed that the model was highly significant. The results of selected experimental degradation conditions matched with the predicted value. FT-IR spectra revealed that the structures of LPS before and after degradation were not significantly changed. Antioxidant activities of LPS revealed that low Mws possessed stronger inhibitory than the original polysaccharides. The scavenging effects on superoxide radicals was the highest when IC50 of crude LPS was 4.92 mg mL^(-1) and IC50 of Mw 18.576 KDa was 1.02 mg mL^(-1), which was fourfold higher than initial polysaccharide.
文摘Effects of fucoidan from Laminaria japonica on 2,4 dinitrochlorobenzene induced delayed type hypersensitivity (DTH) reaction and the serum levels of IgG, IgM, complement C3 and C4 were investigated in the present study. Results showed that oral administration of fucoidan at dose of 150 and 300 mg/(kg·d) for 9 days before the hapten challenge significantly inhibited 2,4 dinitrochlorobenzene induced delayed type hypersensitivity reaction; and also inhibited the humoral immunity. Serum C3 and C4 levels were markedly reduced by fucoidan at dose of 150 and 300 mg/kg; and serum IgG and IgM levels were reduced by fucoidan at dose of 300 mg/kg. The inhibitory effects of fucoidan on delayed type hypersensitivity suggested that it may be potential medication for chronic inflammatory diseases in the future.
文摘FGS was isolated and purified from laminaria japonica by enzymolysis, chromatography methods and so on. The high blood fat model was established by feeding mouse on high fat feedstuff. Compared with control group, the concentration of HDL-C, the ratio of HDL-C/TC increased;while the concentration of TC, LDL-C and TG decreased in experimental groups. Conclusion: FGS was an effective serum lipid regulator and can be developed as medicine and health food.
基金the Technical Program of Jiangsu Province (BE 2006383)the Talent Program of Chinese Academy of Sciences (Dongbei Zhi Chun)
文摘The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments was determined spectroscopically. The results show that: (1) among the six organic solvents, acetone was the most effective one for the extraction; (2) the optimum extraction conditions were as follows: the ratio of S/M (solvent volume/material weight) was 30 ml/g; fresh seaweed was extracted 2 times in 2h; (3) the average total content of pigments was 1.85 mg/g (calculated with dry L. japonica).
