褐藻裙带菜色素—蛋白质复合物的分离与命名

以非离子去污剂癸基－Ｎ－甲基葡萄糖胺为增溶剂,采用聚丙烯酰胺凝胶电泳技术从褐藻裙带菜（Ｕｎｄａｒｉａ ｐｉｎｎａｔｉｆｉｄａ Ｈａｒｖ．）的类囊体膜上分离到８种色素－蛋白质复合物。根据其表观分子量、光谱特性和多肽分析结果,并以高等植物菠菜（Ｓｐｉｎａｃｉａ ｏｌｅｒａｃｅａ Ｌ．）为对照,按照Ａｎｄｅｒｓｏｎ命名系统,８种色素－蛋白质复合物分别命名为ＣＰⅠ ａ、ＣＰⅠ、ＣＰａ、ＬＨＣ１、ＬＨＣ２、     

褐藻裙带菜中PSII复合物及其亚复合物的分离和特性研究

用不连续梯度蔗糖密度超离心,从经TritonX-100增溶的褐藻裙带菜类囊体膜中分离到3种色素蛋白复合物条带,分别是捕光复合物、具有光氧化活性的PSII复合物颗粒(区带II)以及PSI(区带III)。PSII颗粒经毛地黄皂苷增溶后,再次超离心分离得到3条PSII的亚复合物条带。吸收和荧光激发谱显示其中的区带II-1为墨角藻黄素-Chla/c-蛋白复合物,区带II-2为Chla/c-蛋白复合物,两者都只含20kDa多肽;而鲜绿色的区带II-3为不含捕光复合物的活性PSII核心。     

光同化产物在裙带菜（Undaria pinnatifida）中的运转

本文报告了用１４Ｃ标记的光同化产物在裙带菜［Ｕｎｄａｒｉａｐｉｎｎａｔｉｆｉｄａ（Ｈａｒｙ．）Ｓｕｒ］孢子体中的运转现象。观察到光同化产物约需２０ｍｉｎ才能从叶片表皮进入中肋的髓部。在自然条件下,光同化产物主要自叶片梢部经中肋向生长部运转,在叶片梢部和生长部之间存在明显的源一库关系。运转速度约为４２～４８ｃｍ／ｈ。用组织放射自显影的方法证实了髓部的喇叭丝是主要的运转组织。向下运转的溶于酒精的光同化产物中,甘露醇占５０％以上。还在光同化产物中观察到游离的谷氨酸、天门冬氨酸和丙氨酸。     

裙带菜假根中褐藻糖胶的提取

研究了温度、pH值、提取时间和水藻比及添加果胶酶和纤维素酶对采用乙醇沉淀法提取裙带菜假根中褐藻糖胶的影响。单因素和正交实验结果表明,80℃p、H 5、提取时间5 h下的褐藻糖胶得率最高,水藻比影响不明显。添加500 u/g果胶酶和纤维素酶可分别增加57%和46%。     

褐藻裙带菜孢子叶粗提物体外抗单纯疱疹病毒Ⅱ型活性研究

从药物对细胞的保护、对HSV-2增殖的影响及对HSV-2感染细胞的综合作用三个方面研究不同稀释度的裙带菜孢子叶粗提物抑制单纯疱疹病毒Ⅱ型对Vero细胞的感染作用,细胞病变效应法(Cytopathogenic effect,CPE)观察和MTT法测定裙带菜多糖抗HSV-2活性,结果表明裙带菜多糖能明显抑制HSV-2对Vero细胞的致病变作用,使细胞存活率升高,其水提醇沉法所得裙带菜多糖的IC50为6.49μg/mL,并初步推测其抗HSV-2活性是作用在HSV-2和受体结合,侵入Vero细胞阶段,为筛选新型抗病毒药物、研究海藻多糖抗HSV-2活性机理及优化裙带菜孢子叶的提取工艺提供参考依据。     

Temperature requirements for the growth of young sporophytes of Undaria pinnatifida and Undaria undarioides (Laminariales, Phaeophyceae)

The relative growth rate of young sporophytes of Undaria pinnatifida (Harvey) Suringar and Undaria undarioides (Yendo) Okamura was examined in order to understand the difference in distribution of these two species around the coast of Japan. The optimal temperature for growth of both species was similar at 20°C and the upper critical temperature for growth was also similar, at 27°C for U. pinnatifida and 26°C for U. undarioides. Therefore, the optimal and upper critical temperatures for growth of the young sporophytes are not the main factors determining the distribution of each species. Next, the lower critical temperatures for growth were examined. For the young sporophytes of U. pinnatifida, the lower limit was less than 5°C while for those of U. undarioides it was 15°C. Thus, the difference in the lower critical temperature for growth between the two species was approximately 10°C. During the period of young sporophyte growth in the field, the temperature at the mouth of Ise Bay, Japan, where U. pinnatifida occurs, ranges from 12.7°C in December to 13.1°C in April, with a minimum of 7.9°C in February. Our experiments indicate that young sporophytes are able to grow throughout this period. The temperature off Hamajima, Japan, where U. undarioides occurs, ranges from 19.1°C to 14.8°C during the same time period. Again, young sporophytes are able to growth throughout this period, although minimum winter temperatures are only just high enough for growth. These natural temperature ranges during the growth season of the sporophytes agree well with the experimentally determined temperature requirements for growth of each species. Therefore, the difference between the two species in the critical temperature required for growth of the young sporophytes, especially in the low temperature range, is one of the major factors determining the distribution pattern of each species.     

两种电泳系统分离裙带菜色素-蛋白复合物的比较

裙带菜的类囊体膜经过去污剂癸基-N-甲基匍萄糖胺增溶,采用SDS-PAGE分离技术,在Tris-Gly和Tris-硼酸两种电泳系统中分离其色素-蛋白质复合物,并比较其复合物的光谱特性。结果表明:采用Tris-Gly电泳分离系统从裙带菜中分离到8种色食-蛋白质复合物,分别是CP Ia、CPI、LHC₁、CPa、LHC₂、LHC₃、LHC₄和LHC₅。在Tris-硼酸电泳分离系统中共分离到5种色素-蛋白质复合物,分别是CPI、CPa、LHC₁、LHC₂、LHC₃。吸收光谱和荧光光谱的测定结果表明,两种电泳系统中分离的相对应条带的光谱特性基本相近。     

The effect of fertilizer application on farming of the seaweed Undaria pinnatifida (Laminariales, Phaeophyta)

A slow-release ammonium phosphate fertilizer coated with porous plastic was tested on Undaria pinnatifida (Harvey) Suringar as a possible solution to the nutrient deficiency in seawater that causes quality and yield deterioration in seaweed farming. The yield of U. pinnatifida within the fertilized area was 17–40% greater than that of the control area (unfertilized area). In addition, two harvests were possible per season and the quality of harvested U. pinnatifida was also better than that outside the fertilizer diffusion area. The released NH₄-N did not increase the concentration of NH₄-N outside the farming area. Therefore, this fertilizer increased yield and improved quality without causing water pollution.     

Responses of vegetative gametophytes of Undaria pinnatifida to high irradiance in the process of gametogenesis

The population of Undaria pinnatifida in its ecologic niche sustains itself in high temperature summer in the form of vegetative gametophytes, the haploid stage in its heteromorphic life cycle. Gametogenesis initiates when seawater temperature drops below the threshold levels in autumn in the northern hemisphere. Given that the temperature may fall into the appropriate range for gametogenesis, the level of irradiance determines the final destiny of a gametophytic cell, either undergoing vegetative cell division or initiating gametogenesis. In elucidating how vegetatively propagated gametophytes cope with changes of irradiance in gametogenesis, we carried out a series of culture experiments and found that a direct exposure to irradiance as high as 270 μmol photons m^?2 s^?1 was lethal to dim‐light (7–10 μmol photons m^?2 s^?1) adapted male and female gametophytes. This lethal effect was linearly corelated with the exposure time. However, dim‐light adapted vegetative gametophytes were shown to be able tolerate as high as 420 μmol photons m^?2 s^?1 if the irradiance was steadily increased from dim light levels (7–10 μmol photons m^?2 s^?1) to 90, 180 and finally 420 μmol photons m^?2 s^?1, respectively, at a minimum of 1–3 h intervals. Percentage of female gametophytic cells that turned into oogonia and were eventually fertilized was significantly higher if cultured at higher but not lethal irradiances. Findings of this investigation help to understand the dynamic changes of population size of sporophytic plants under different light climates at different site‐specific ecologic niches. It may help to establish specific technical details of manipulation of light during mass production of seedlings by use of vegetatively propagated gametophytes.     

Temperature requirements for the growth and maturation of the gametophytes of Undaria pinnatifida and U. undarioides (Laminariales, Phaeophyceae)

Gametophytes of two Undaria species, U. pinnatifida and U. undarioides (Laminariales, Phaeophyceae), were studied to determine their water temperature requirements in order to understand their different distributions in Mie Prefecture, Japan. The optimal temperature for growth was 20°C for gametophytes of both species, and the upper critical temperature for growth was also the same for both species at 28°C. Therefore, the optimal and critical temperatures for growth of the gametophytes are not the main factors determining distribution. The optimal temperature for maturation of U. pinnatifida was approximately 10–15°C, whereas it was closer to 20–21°C for U. undarioides, a difference between these species of at least 5°C. In autumn and early winter, the seawater temperature at the mouth of Ise Bay, where U. pinnatifida is distributed, ranges from 21.6°C (October) to 12.7°C (December), and off Hamajima, where U. undarioides is found, the range is from 22.7°C (October) to 19.1°C (December). The seawater temperatures from October to December, which is the maturation season for the gametophytes, agreed well with the optimal temperature requirements for maturation of the gametophytes of both species. Thus the difference in the maturation temperature range of the gametophytes is a major factor determining distribution of these Undaria species along the Japanese coast.     

裙带菜褐藻糖胶的分离纯化及其结构分析

裙带菜经水提法提取得到褐藻糖胶粗多糖,经DEAE-Sepharose FF离子交换层析和 Sepharose 4B层析后,得到Sl、S2两个单一组分,相对分子质量分别为550 808、38 335.基本结构及单糖组成分析表明,二者均含有岩藻糖、糖醛酸、硫酸基、半乳糖、甘露糖、葡萄糖、鼠李糖、木糖、阿拉伯糖,但含量差别较大,推测与二者的生理活性的差异有关.     

Novel means for variety breeding and sporeling production in the brown seaweed Undaria pinnatifida (Phaeophyceae): Crossing female gametophytes from parthenosporophytes with male gametophyte clones

The unialgal haploid gametophyte clones are frequently used for variety breeding and sporeling production in the brown seaweed Undaria pinnatifida because a single crossing of a pair of the selected male and female gametophyte clones can generate sporophytic offspring with identical genotype and phenotype. As the seeds to be sprayed on the collectors, the detachment rate of the filamentous gametophytes is high in comparison to the seeded spores. In this investigation, we report the use of parthenogenesis to achieve the same goal in variety selection and sporeling production but with higher efficiency. The selected female unialgal gametophyte clone (Code: 06‐8‐1F) was induced to produce parthenosporophytes. These sporophytes were grown up in a controlled system and used to release zoospores. All zoospores generated into female gametophytes. These female gametophytes were allowed to go through parthenogenesis for the second year to confirm the applicability of this means. In the third year, the zoospores released from the parthenosporophytes were seeded on collectors over summer in female gametophyte form. In the early autumn, a selected male unialgal gametophyte clone (code: 5#F1‐2‐5M) was used to cross the seeded female gametophytes on the collectors. When the sporelings reached a mean length of 780 μm, they were transplanted to open sea on longlines for growing up. At harvest, the average length, width and wet weight of the adult sporophytes were 211 cm, 48.8 cm and 373 g, respectively. The sporophytic blades were uniformly smooth without wrinkles on both sides of the midrib, indicating top quality of the products. Amplified fragment length polymorphism analyses confirmed the identical genotypes of sporophytic offspring. These results suggested that this novel variety breeding and sporeling production method could serve as an efficient alternative to the traditional breeding technique for U. pinnatifida and possibly other commercial kelps that have identical life cycles.     

Sex‐linked microsatellite marker detected in the female gametophytes of Undaria pinnatifida (Phaeophyta)

In our microsatellite analysis of three male and three female gametophytes of Undaria pinnatifida (Harv.) Suringar, a microsatellite marker (part of the locus Up‐AC‐2A8, GenBank accession no. AY738602.1) was only polymerase chain reaction‐amplified in three female gametophytes. This putative female‐specific marker was further tested by the use of 32 male and 21 female gametophytes maintained in the Marine Biological Culture Collection Centre, China. In addition, three sporophytes were included for confirmation. Results showed that the marker was present in all of the female gametophytes and sporophyte cultures, but absent in all of the male gametophytes. To our knowledge, this is the first sex‐related marker ever reported in U. pinnatifida. The discovery of this marker will accelerate gender identification and shed light on our understanding of the mechanisms of sex determination at a molecular level in this commercially important seaweed.     

Variation in metal concentrations in the brown alga Undaria pinnatifida in Osaka Bay, Japan

In order to evaluate the usefulness of a biomonitoring system using seaweeds for assessing the geographic distribution of metal ions in coastal seawaters, the metal concentrations in the sporophytes of an annual kelp, Undaria pinnatifida (Harvey) Suringar, were collected at 15 localities in Osaka Bay, Japan and compared. About 160 cm² of the blade was cut out from the central part of clean sporophytes, rinsed in filtered seawater using an ultrasonic cleaning bath, and freeze‐dried. After digestion with 12% HNO₃ in a microwave apparatus, metal concentrations in the samples were determined by inductively coupled plasma mass spectrometry. The concentrations (dry weight basis) of most examined elements were in the parts per million range; Cr, 0.48–3.18; Ni, 0.77–5.94; Cu, 3.20–43.8; Zn, 11.3–86.8; Pb, 0.14–3.53. Comparisons of metal compositions of the U. pinnatifida samples from the northeastern area of the bay, which has a large urban population and highly developed industries inland, showed high concentrations of Cd, Pb and Cu compared with the samples from the southwestern area of the bay where the population and industries are much smaller. This suggests that U. pinnatifida metal loads can be used as a marker to track the geographic distributions of the metal concentrations in coastal seawaters, reflecting inland human activities such as shipbuilding and repairing in port areas, and can be used as a useful biomonitoring system of coastal environments for long‐term trend.     

Functional restoration of replicative senescent mesenchymal stem cells by the brown alga Undaria pinnatifida

The brown alga Undaria pinnatifida, which is called Mi-Yoek in Korea, has been traditionally consumed as a health food in East Asian countries. Recent studies have reported that U. pinnatifida has beneficial effects on arteriosclerosis, inflammation, fat metabolism, and tumors. In this study, we examined the anti-senescence effects of ethanol extracts of U. pinnatifida (UP-Ex) in human bone marrow mesenchymal stem cells (hBM-MSCs). UP-Ex protected hBM-MSCs against oxidative injury, as determined by MTT assays. This effect was confirmed by immunoblot analysis of the oxidation-sensitive protein p53 and the apoptotic protein cleaved caspase-3. Excessive intracellular reactive oxygen species (ROS) accumulation induced by oxidative stress was moderated in UP-Ex-treated hBM-MSCs (UP-Ex-MSCs). Similarly, expression of the ROS-scavenging enzymes superoxide dismutase 1 (SOD1), SOD2, and catalase was recovered in UP-Ex-MSCs. Excessive ROS induced by long-term cell expansion (passage 17) was significantly decreased along with restoration of the senescence proteins p53, p21, and p16 in UP-Ex-MSCs. UP-Ex treatment also improved the ability of these replicative, senescent hBM-MSCs (passage 17) to differentiate into osteocytes or adipocytes, suggesting that UP-Ex ameliorates the functional decline of senescent stem cells and may provide better therapeutic efficacy in stem cell therapy.

Abbreviations: hBM-MSCs: human bone marrow mesenchymal stem cells; DCF: 2′,7′-dichlorodihydrofluorescein; DCFH-DA: 2′,7′-dichlorofluorescein diacetate; MTT: 3-(4,5-dimethylthiazol-2-yl-)2,5-diphenyltetrazolium bromide; PBS: phosphate-buffered saline; PFA: paraformaldehyde; RIPA: radioimmunoprecipitation assay; ROS: reactive oxygen species; SOD1: superoxide dismutase 1; SOD2: superoxide dismutase 2.     

Distribution of introduced Japanese macroalgae Undaria pinnatifida,Sargassum muticum (Phaeophyta) and Antithamnion Pectinatum (Rhodophyta) in the Lagoon of Venice

The present distribution of invasive seaweeds Undaria pinnatifida (Phaeophyta, Laminariales) and Sargassum muticum (Phaeophyta, Fucales) in the Lagoon of Venice is updated in this work. The distribution of Antithamnion pectinatum (Rhodophyta, Ceramiales) is presented for the first time. All these species have quickly colonized the hard substrata competing with indigenous species. Competition acts differently in the two species, U. pinnatifida competing for the substratum and S. muticum for light. On tidal flats composed of silt and sand, the growth of two brown seaweeds is restricted owing to the lack of a solid substrata. While Undaria pinnatifida and Sargassum muticum have colonized only some areas of the lagoon in a short vertical range (from 0 to -3 m), Antithamnion pectinatum is present on the three portmouths, along the inner shores of the city of Venice, and in the lagoon islands with extensive vertical distribution (from +0.2 to -8 m). This revised version was published online in September 2006 with corrections to the Cover Date.     

The enhancing effect of fucoidan derived from Undaria pinnatifida on immunoglobulin production by mouse spleen lymphocytes

In this study, we revealed that a Mekabu (Udaria pinnantifida) extract enhanced immunoglobulin (Ig) production of mouse spleen lymphocytes. Furthermore, it was suggested that water-soluble and high molecular weight ingredients in the Mekabu extract have significant enhancing effect on Ig production. Therefore, fucoidan was estimated as the active component.     

Assessing genetic identity of sporophytic offspring of the brown alga Undaria pinnatifida derived from mono‐crossing of gametophyte clones by use of amplified fragment length polymorphism and microsatellite markers

The haploid stage of gametophytes of the subtidal brown alga Undaria pinnatifida can be vegetatively propagated under favorable conditions. This unique characteristic makes it possible to establish independent gametophyte cell lines that are zoospore‐derived. Sporophytic offspring can be generated through hybridizing the male and female gametophytes, which are derived from different cell lines. Accumulated experiences in this and other species in Laminariales demonstrated the applicability of this novel way to breed desired strains for open‐sea cultivation. Sporophytic offspring originated from mono‐crossing of male and female gametophyte clones were shown to have similar morphological characteristics under identical ambient conditions. However, there has been no report to relate this similarity on molecular levels. In this report, amplified fragment length polymorphism (AFLP) and microsatellite markers were used to analyze the genetic identity of sporophytic offspring of U. pinnatifida originated from two mono‐crossing lines (M1 and M2), two self‐breeding lines (S1 and S2) and one wild population (W). Totally 318 AFLP loci were revealed by use of 11 primer sets, of which 4.7%, 0.3%, 17.9%, 16.4% and 36.5% were polymorphic in M1, M2, S1, S2 and W, respectively. The pairwise genetic identity among the individuals of the same line was assessed. It was shown that offspring from mono‐crossing lines had a higher degree of identity (95.6–100%) than self‐breeding lines (87.7–98.4%) and the wild population (81.5–92.1%). Analysis by use of six microsatellite loci also revealed a higher genetic identity among individuals of the mono‐crossing line, further confirming the results of AFLP analysis. Results from this investigation support, on molecular levels, the novel way to produce and maintain strains in U. pinnatifida by use of different gametophyte cell lines.     

Cultivation and utilization of Undaria pinnatifida (wakame) as food

A brief review is presented concerning wakame, Undaria pinnatifida, one of the most popular seaweeds used for food in Japan. Although it has been cultivated since about 1940, full-scale cultivation occurred after 1955. As methods for providing ‘seed stock’ and of processing the harvested sporophytes progressed, the yield increased rapidly. The main areas of cultivation are in Japan (e.g. Sanriku, Naruto), Korea and China, while ‘wild’ U. pinnatifida has been introduced into France, New Zealand and Australia. The total world yield of wakame exceeds 500 000 t fresh weight. Cultivated and harvested Undaria is boiled and salted (thus becoming green) and refrigerated; in the factory, it is removed its foreign matter and salt and dried. After checking for quality, the product is packaged in forms convenient for cooking and eating.     

Seasonal growth,erosion rates,and nitrogen and photosynthetic ecophysiology of Undaria pinnatifida (Heterokontophyta) in southern New Zealand1

The goal of this study was to examine growth, erosion rates, and the photosynthetic and nitrogen ecophysiology of the invasive seaweed Undaria pinnatifida (Harv.) Suringar. Sporophytes of U. pinnatifida that appeared in Otago Harbour, southeastern New Zealand, in late autumn (May) 1996 were tagged, and their growth rates followed until the onset of senescence in early summer (November 1996). Blade growth rates were maximal between May and August when they ranged from 0.77 ± 0.05 to 0.93 ± 0.05 cm · d^?1 and declined from September onward. In laboratory experiments, U. pinnatifida took up nitrate and ammonium simultaneously at rates ranging from 21.3 ± 2.1 to 179.3 ± 65.1 μmol · g^?1 dry weight (dwt) · h^?1. When monthly patterns of growth rate, seawater inorganic nitrogen, nitrogen uptake kinetics, soluble tissue nitrate, % tissue carbon (C) and nitrogen (N), and C:N ratio were considered together, there was no evidence that N limited the growth of U. pinnatifida. Furthermore, the photosynthetic parameters P_max, α, E_k, E_c, and R_d derived from P versus E curves indicated that the growth of U. pinnatifida was not light limited, and that the population could potentially grow deeper than its observed location at 4 m depth. Nitrogen and light ecophysiological parameters of U. pinnatifida more closely resemble those of small, ephemeral seaweeds, such as Ulva, than other members of the Laminariales. We suggest that a “plastic” physiology may allow U. pinnatifida sporophytes to match their physiology to a range of environments, which is one reason for its success as an invasive seaweed.