Breeding practice for Undaria pinnatifida （Harvey） Suringar requires the screening of a large number of offspring from gametophyte crossings to obtain an elite variety for large-scale cultivation. To better understa...Breeding practice for Undaria pinnatifida （Harvey） Suringar requires the screening of a large number of offspring from gametophyte crossings to obtain an elite variety for large-scale cultivation. To better understand the genetic relationships of different gametophyte cultures isolated from different sources, 20 microsatellite loci were screened and 53 gametophyte clone cultures analyzed for U. pinnatijqda isolated from wild sporophytes in Vladivostok, Russia and from cultivated sporophytes from Dalian and Qingdao, China. One locus was abandoned because of poor amplification. At the sex-linked locus of Up-AC-2AS, 3 alleles were detected in 25 female gametophyte clones, with sizes ranging from 307 to 316 bp. At other loci, 3 to 7 alleles were detected with an average of 4.5 alleles per locus. The average number of alleles at each locus was 1.3 and 3.7 for Russian and Chinese gametophyte clones, respectively. The average gene diversity for Russian, Chinese, and for the combined total of gametophyte clones was 0.1, 0.4, and 0.5, respectively. Russian gametophyte clones had unique alleles at 7 out of the 19 loci. In cluster analysis, Russian and Chinese gametophyte clones were separated into two different groups according to genetic distance. Overall, high genetic diversity was detected in gametophyte clones isolated from the two countries. These gametophyte cultures were believed to be appropriate parental materials for conducting breeding programs in the future.展开更多
In our trials, from 2007 to 2008, of mass production of seedlings of Hizikiafusiformis using synchronization techniques, problems of a ＂dark thalli＂ phenomenon and epiphytes contamination severely threatened the hea...In our trials, from 2007 to 2008, of mass production of seedlings of Hizikiafusiformis using synchronization techniques, problems of a ＂dark thalli＂ phenomenon and epiphytes contamination severely threatened the health of juvenile seedlings. In this investigation, we optimized conditions for improving the growth of juvenile seedlings. Seven string collectors were seeded with zygotes and a series of experiments were conducted including direct exposure to solar irradiance, co-culture with Ulva spp. and treatment with sodium hypochlorite. It was found that direct exposure to solar irradiance （maximum： 1 740 μmol photons/（m2.s）） for 2 h per day could efficiently enhance the growth of young seedlings and simultaneously inhibit the growth of epiphytic algae. In this treatment, 50-day old seedlings could reach an average of 0.44 cm in length and possess up to nine leaflets. However, a single treatment with 18-mmol/L sodium hypochlorite for 10 rain severely harmed 15-day old seedlings. In comparison, weekly treatment with 2.2-mmol/L sodium hypochlorite for 10 rain brought no apparent harm to seedlings and eliminated epiphytic algae efficiently. However, this treatment significantly increased the detachment rate of seedlings, Inoculating Ulva spp. onto the collector caused a dramatic decrease in the number of seedlings. However, the growth of the remaining seedlings appeared unhampered. All collectors except the control were daily sprayed with a high pressure water jet from the 84 day post fertilization. From the first day to 50th day, no ＂dark thallus＂ was observed on any of the seven collectors. We believe that well-timed daily exposure to solar irradiance would favor H. fusiformis in its early growing stages.展开更多
The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived mono...The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.展开更多
Sargassum muticum is one of the most abundant and conspicuous native macroalgae species on the northern coasts of China. It often forms large-scale seaweed beds in subtidal zones. This investigation was designed to un...Sargassum muticum is one of the most abundant and conspicuous native macroalgae species on the northern coasts of China. It often forms large-scale seaweed beds in subtidal zones. This investigation was designed to understand the intraspecific genetic relationships of this alga based on samples collected from four northern coastal sites of China, and to evaluate gamete release and growth capacity in laboratory conditions. The nuclear internal transcribed spacer 2 （ITS2） sequences of 16 samples from four locations were identical. Based on cox3 gene and partial rbcLS operon sequences, intraspecific genetic variability was detected with three and two ribotypes, respectively. Temperature, not irradiance, was shown to significantly affect gamete release and fertilization. Elevated temperature and irradiance enhanced the growth of germlings and vegetative branchlets. Maximum growth rate of germlings was detected at 18-24℃and an irradiance of 60-100 μmol photons/（m^2.s）. Under ambient conditions （12-25℃ and 60-125 gmol photons/ （m2.s））, relative growth rate of young branchlets could reach 7.5%/d.展开更多
During sexual reproduction of seaweeds, spermatozoid (sperm) discharge is triggered by chemical messengers (pheromones) released by the female gametes. The chemotactic ability of the sperm ensures fertilization succes...During sexual reproduction of seaweeds, spermatozoid (sperm) discharge is triggered by chemical messengers (pheromones) released by the female gametes. The chemotactic ability of the sperm ensures fertilization success. Using unialgal male and female gametophyte material under designated standard gametogenesis testing (SGT) conditions, the potential life-span of the sperm of two seaweeds, Saccharina japonica and Undaria pinnatifida, was assessed by their ability to fertilize eggs. Results show that within 20-30 min after being discharged, sperm of both species could complete fertilization without an apparent decline in fertilization rate. Although fertilization rate 60-120 min after sperm discharge dropped significantly in both species, some sperm were viable enough to fertilize the eggs. In S.japonica, at 12°C, some sperm were able to fertilize eggs up to 12 h after discharge. In both species, egg discharge rates (EDR) in the male and female mixed positive controls were significantly higher than those of all the sperm-testing groups. Doubling the seeded male gametophytes of S. japonica in the SGT tests significantly increased the EDR, further confirming the effect of the presence of the male on the female in terms of facilitating egg discharge from oogonia.展开更多
During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic ...During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic （18S rDNA and 16S rDNA sequences） analyses revealed a close affinity with Eutreptiella gymnastica, a bloom-forming species. Different culture conditions were monitored to understand optimal E. gymnastica growth characteristics. The optimal growth conditions in a batch culture of this isolate were 20C, 160 gmol photons/（mE.s） of white light, and a salinity of 10-31. Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 gmol/L. Understanding the effect of culture conditions on E. gymnastica may help understanding the blooming mechanism of this alga in its natural environment.展开更多
基金Supported by the National Special Research Fund for Non-Profit Sector(Agriculture)(No.200903030)the Agriculture Science Technology Achievement Transformation Fund(No.2010GB24910699)the Knowledge Innovation Programs of Chinese Academy of Sciences(No.KSCX2-EW-B-3)
文摘Breeding practice for Undaria pinnatifida （Harvey） Suringar requires the screening of a large number of offspring from gametophyte crossings to obtain an elite variety for large-scale cultivation. To better understand the genetic relationships of different gametophyte cultures isolated from different sources, 20 microsatellite loci were screened and 53 gametophyte clone cultures analyzed for U. pinnatijqda isolated from wild sporophytes in Vladivostok, Russia and from cultivated sporophytes from Dalian and Qingdao, China. One locus was abandoned because of poor amplification. At the sex-linked locus of Up-AC-2AS, 3 alleles were detected in 25 female gametophyte clones, with sizes ranging from 307 to 316 bp. At other loci, 3 to 7 alleles were detected with an average of 4.5 alleles per locus. The average number of alleles at each locus was 1.3 and 3.7 for Russian and Chinese gametophyte clones, respectively. The average gene diversity for Russian, Chinese, and for the combined total of gametophyte clones was 0.1, 0.4, and 0.5, respectively. Russian gametophyte clones had unique alleles at 7 out of the 19 loci. In cluster analysis, Russian and Chinese gametophyte clones were separated into two different groups according to genetic distance. Overall, high genetic diversity was detected in gametophyte clones isolated from the two countries. These gametophyte cultures were believed to be appropriate parental materials for conducting breeding programs in the future.
基金Supported by the National High Technology Research and Development Program of China (863 Program) (Nos. 2006AA10A412 2006AA10A416)+1 种基金Main Program of National Science Infrastructure Platform, a project from the Ministry of Science and Technology of China (No. 2006DKA30470-017)a non-profit program from the Ministry of Agriculture of China (No. 200903030)
文摘In our trials, from 2007 to 2008, of mass production of seedlings of Hizikiafusiformis using synchronization techniques, problems of a ＂dark thalli＂ phenomenon and epiphytes contamination severely threatened the health of juvenile seedlings. In this investigation, we optimized conditions for improving the growth of juvenile seedlings. Seven string collectors were seeded with zygotes and a series of experiments were conducted including direct exposure to solar irradiance, co-culture with Ulva spp. and treatment with sodium hypochlorite. It was found that direct exposure to solar irradiance （maximum： 1 740 μmol photons/（m2.s）） for 2 h per day could efficiently enhance the growth of young seedlings and simultaneously inhibit the growth of epiphytic algae. In this treatment, 50-day old seedlings could reach an average of 0.44 cm in length and possess up to nine leaflets. However, a single treatment with 18-mmol/L sodium hypochlorite for 10 rain severely harmed 15-day old seedlings. In comparison, weekly treatment with 2.2-mmol/L sodium hypochlorite for 10 rain brought no apparent harm to seedlings and eliminated epiphytic algae efficiently. However, this treatment significantly increased the detachment rate of seedlings, Inoculating Ulva spp. onto the collector caused a dramatic decrease in the number of seedlings. However, the growth of the remaining seedlings appeared unhampered. All collectors except the control were daily sprayed with a high pressure water jet from the 84 day post fertilization. From the first day to 50th day, no ＂dark thallus＂ was observed on any of the seven collectors. We believe that well-timed daily exposure to solar irradiance would favor H. fusiformis in its early growing stages.
基金Supported by the National Natural Science Foundation of China(Nos.41176135,41206142)the Innovation Program of Chinese Academy of Sciences(No.KSCX2-EW-B-3)the Ministry of Agriculture of China(No.200903030)
文摘The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A413)the National Natural Science Foundation of China(Nos.41206146,41176135)the Leading Academic Discipline Project of Shanghai Municipal Education Commission(No.J50701)
文摘Sargassum muticum is one of the most abundant and conspicuous native macroalgae species on the northern coasts of China. It often forms large-scale seaweed beds in subtidal zones. This investigation was designed to understand the intraspecific genetic relationships of this alga based on samples collected from four northern coastal sites of China, and to evaluate gamete release and growth capacity in laboratory conditions. The nuclear internal transcribed spacer 2 （ITS2） sequences of 16 samples from four locations were identical. Based on cox3 gene and partial rbcLS operon sequences, intraspecific genetic variability was detected with three and two ribotypes, respectively. Temperature, not irradiance, was shown to significantly affect gamete release and fertilization. Elevated temperature and irradiance enhanced the growth of germlings and vegetative branchlets. Maximum growth rate of germlings was detected at 18-24℃and an irradiance of 60-100 μmol photons/（m^2.s）. Under ambient conditions （12-25℃ and 60-125 gmol photons/ （m2.s））, relative growth rate of young branchlets could reach 7.5%/d.
基金Supported by National High Technology Research and Development Program of China (863 Program)(No.2012AA10A413)the National Natural Science Foundation of China (Nos.41206146,41176135,41206142)+3 种基金the National Special Research Fund for Non-Profit Sector (Agriculture) from the Ministry of Agriculture (No.200903030)the Knowledge Innovation Program of Chinese Academy of Sciences (No.KSCX2-EW-B-3)the National Key Basic Program of Science and Technology-Platforms of Aquaculture Stock Resourcesthe Leading Academic Discipline Project of the Shanghai Municipal Education Commission (No.J50701)
文摘During sexual reproduction of seaweeds, spermatozoid (sperm) discharge is triggered by chemical messengers (pheromones) released by the female gametes. The chemotactic ability of the sperm ensures fertilization success. Using unialgal male and female gametophyte material under designated standard gametogenesis testing (SGT) conditions, the potential life-span of the sperm of two seaweeds, Saccharina japonica and Undaria pinnatifida, was assessed by their ability to fertilize eggs. Results show that within 20-30 min after being discharged, sperm of both species could complete fertilization without an apparent decline in fertilization rate. Although fertilization rate 60-120 min after sperm discharge dropped significantly in both species, some sperm were viable enough to fertilize the eggs. In S.japonica, at 12°C, some sperm were able to fertilize eggs up to 12 h after discharge. In both species, egg discharge rates (EDR) in the male and female mixed positive controls were significantly higher than those of all the sperm-testing groups. Doubling the seeded male gametophytes of S. japonica in the SGT tests significantly increased the EDR, further confirming the effect of the presence of the male on the female in terms of facilitating egg discharge from oogonia.
基金Supported by the National Special Research Fund for Non-Profit Sector(Agriculture) (No. 200903030)the National Key Basic Program of Science & Technology-Platforms of Aquaculture Stock Resourcesa Project from Chinese Academy of Sciences
文摘During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic （18S rDNA and 16S rDNA sequences） analyses revealed a close affinity with Eutreptiella gymnastica, a bloom-forming species. Different culture conditions were monitored to understand optimal E. gymnastica growth characteristics. The optimal growth conditions in a batch culture of this isolate were 20C, 160 gmol photons/（mE.s） of white light, and a salinity of 10-31. Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 gmol/L. Understanding the effect of culture conditions on E. gymnastica may help understanding the blooming mechanism of this alga in its natural environment.
