Anti-obesity effects of fucoidan from Sargassum thunbergii in adipocytes and high fat diet induced obese mice through inhibiting adipogenic specific transcription factor

The prevalence of obesity has increased and is a health concern worldwide.Due to the concerns regarding synthetic anti-obesity treatments,nowadays natural products become a trend.Previous studies proved that there is a potential to use marine algae as anti-obesity agents.Therefore,in this study,the lipid inhibitory effect of crude polysaccharide of amyloglucosidase-assisted hydrolysate from Sargassum thunbergii(STAC)and its fucoidan fractions(STAFs)on 3T3-L1 cells and high-fat diet(HFD)-induced obese mice were investigated.According to the results,the STAF3,showed the highest xylose content and exhibited significant inhibitory effects on lipid accumulation by downregulating adipogenic and lipogenic proteins in 3T3-L1 cells.Furthermore,oral supplementation with STAC significantly declined gain in body weight and fat weight,and serum lipid contents in an HFD-induced obesity mouse model.Structural and chemical characterizations demonstrated that puritied STAF3 has consistent surface morphology and small particle size,with similar structural characteristics as commercial fucoidan.Together,these results indicate that STAC and purified STAF3 from Sargassum thunbergia is a potent source to develop as ananti-obesity agents or functional food products to counter obesity.

Effects of low molecular weight polysaccharide from Sargassum thunbergii against palmitic acid-induced intracellular lipid accumulation in 3T3-L1 adipocyte and HepG2 cells

Low molecular weight polysaccharides can be isolated from Sargassum thunbergii(LMPST)and in vitro experiments were conducted to evaluate the inhibitory effects on lipids.Two natures of LMPST were attained from S.thunbergii and appraised their LMPST on palmitic acid(PA)induced lipid accretion in Hep G2,and 3T3-L1 cells.LMPST treatment lessened lipid deposition and intracellular free fatty acid and triglyceride intensities in PA-treated above mentioned cells.The mechanistic study publicized that LMPST2 significantly suppressed adipogenesis and stimulated the PA-treated 3T3-L1 cells occupied in the lipolysis pathway.Furthermore,in PA-treated Hep G2 cells,the free fatty acid oxidation was significantly increased by LMPST2.Given these constructive properties of LMPST2 from S.thunbergii,is a potential candidate for diminishing the intracellular lipids,and for a therapeutic agent in those conditions.

Effects of Different Seed Stem Sizes on the Changes of Available Elements in Rhizosphere Soil of Fritillaria thunbergii Miq.

[Objectives]This study was conducted to screen suitable seed stems of Fritillaria thunbergii Miq.from different provenances and to provide a theoretical basis for the high-yielding and high-efficiency cultivation of F.thunbergii Miq.introduced to different places.[Methods]F.thunbergii Miq.from four different provenances including Zhejiang,Nantong and Chongqing were introduced and cultivated in Wanzhou of Chongqing.The contents of available Zn,Fe,Mn,Cu,Mo,N,P,K,Ca and Mg in rhizosphere soil of F.thunbergii Miq.during five growing stages were determined after selecting different stem sizes for field cultivation.[Results]Small stems of Pan an and Ningbo provenances(SSG3,121-160/kg)and middle stems of Nantong and Fengjie provenances(SSG2,81-120/kg)showed higher soil availability.[Conclusions]In the process of introduction and cultivation of F.thunbergii Miq.,high yield and high efficiency can be achieved by selecting smaller seed stems of F.thunbergii Miq.

Polyphenols from Vitis thunbergii Sieb. et Zucc.

Aim Isolation and structural elucidation of the constituents from the aerial part of Vitis thunbergii . Methods To isolate chemical constituents, column chromatography and HPLC were used. Physico chemical characterization and spectroscopic analysis were employed for structural identification. Results Eleven polyphenols were isolated and identified. Conclusion Compound 1 is a new compound and its structure was characterized to be 3,5 dimethoxyl 4 hydroxyl phenylpropanol 9 O β D glycopyranoside.

鼠尾藻(Sargassum thunbergii)有性生殖过程与育苗

采用室内培养和野外实地观测的方法,研究了鼠尾藻的生长发育和有性生殖过程,并在此基础上进行了有性生殖人工育苗实验。结果表明,鼠尾藻在海水温度高于22℃时,生殖托开始发育,海水温度超过23℃以后,生殖托的生长速度加快,海水温度超过24℃时,生殖托生长速度最快并迅速成熟,在海水温度达到26℃时,观察到了卵的集中排放挂托现象。鼠尾藻卵的排放较为齐整且有一定的先后顺序,生殖托基部的卵最先排出,中部次之,最后是顶部,雄托排精时间要稍晚于雌托排卵时间。受精卵大约在受精1h后进行第一次细胞分裂,分裂面与长轴垂直,大约2h后进行第二次分裂,下端细胞的分裂面平行于第一次分裂方向,而上端细胞的分裂面则垂直,随后大约每2—4h细胞分裂一次。从卵挂托到幼胚脱落一般需要24—48h。观察发现,生殖窝不仅存在于生殖托上,在部分生殖枝上也有分布。在鼠尾藻的人工育苗的实验中,胚苗在苗帘上的生长状态良好。

我国沿海鼠尾藻(Sargassum thunbergii)遗传资源RAPD分析

采用RAPD分子标记技术分析了我国沿海五个海区(大连、威海、青岛、温州、福州)鼠尾藻种群的遗传多样性。从60条随机引物中筛选出21条引物,对五个鼠尾藻种群共150份DNA样本进行了扩增,共检测到207个有效位点,多态位点达89.5%,Nei基因多样性指数(H)为0.2966,Shannon信息指数(I)为0.4508;种群间的遗传分化系数(Gst)为0.6131,基因流(Nm)为0.3155。AMOVA分析表明Fst为0.6185。我国沿海鼠尾藻种群遗传多样性较低,遗传分化水平较高。

桑沟湾筏式养殖鼠尾藻(Sargassum thunbergii)的生长特性

2014年4–6月在桑沟湾海区进行鼠尾藻(Sargassum thunbergii)海上筏式养殖实验,分析了鼠尾藻在桑沟湾的生长特性,调查了藻体上的附着生物。结果显示,(1)鼠尾藻在桑沟湾海域生长迅速,水温为10–17℃时特定生长率最高,可达6.10%/d;根据特定生长率与水温的关系,获得了鼠尾藻的最佳生长温度为14.9℃;(2)5月10日开始有生殖托形成,水温达到20.4℃时,鼠尾藻生殖托大量成熟,并有放散;(3)养殖期间,鼠尾藻最大长度达187.05 cm,均长可达112.31 cm;干湿比由0.147(4月)上升至0.189(6月);每公顷产量可达43.95 t(湿重),相当于干重为8.25 t;(4)藻体上有大型附着生物16种,主要优势种为尖嘴扁颌针鱼鱼卵、玻璃海鞘和海绵;附着生物的生物量随着水温升高而增加。研究表明,海区的附着生物对鼠尾藻的生长影响不大,在桑沟湾大规模养殖鼠尾藻是可行的。

Improvement of the zygote utilization and reduction of the seedling loss in the early stage of seedling production of Sargassum thunbergii (Fucales, Phaeophyta)

Artificial seedling production of Sargassum thunbergii is an effective way to relieve pressure on natural resources.In order to improve the utilization of zygotes and reduce the loss of seedlings,studies on the characteristic of the zygotes release,the development of rhizoids,the attachment of germlings,and the influence of jet washing were conducted.Results show that the percent of zygotes released was increased with time in the first 60 h.The capacity of germlings attached to the substratum was significantly increased,especially coincident with the time of the new rhizoids emerged and elongated.The detachment rate of germlings significantly decreased with the delay of starting time of jet washing or the reduction of jet washing velocity.However,the jet washing at any level applied in the experiment could cause considerable loss of germlings within the 20 days after the attachment.Our study provided some parameters to optimize the operation in the early stage of seedling production.

Effects of Ion Implantation on in Vitro Pollen Germination and Cellular Organization of Pollen Tube in Pinus thunbergii Parl. (Japanese Black Pine)

Low-energy ion implantation, as a new technology to produce mutation in plant breeding, has been widely applied in agriculture in China. But so far there is a little understanding of the underlying mechanisms responsible for its biological effects at the cellular level. Here we report the biological effects of a nitrogen ion beams of 30 keV on the pollen grains of Pinus thunbergii Parl. In general, ion implantation inhibited pollen germination. The dose-response curve presented a particular saddle-like pattern. Ion implantation also changed the dimension of the elongated tubes and significantly induced tip swelling. Confocal microscopy indicated that the pollen tube tips in P. thunbergii contained an enriched network of microtubules. Ion implantation led to the disruption of microtubules especially in swollen tips. Treatment with colchicine demonstrated that tip swelling was caused by the disruption of microtubules in the tip, indicating a unique role for microtubules in maintaining the tip integrality of the pollen tube in conifer. Our results suggest that ion implantation induce the disruption of microtubule organization in pollen and pollen tubes and subsequently cause morphological abnormalities in the pollen tubes. This study may provide a clue for further investigation on the interaction between low-energy ion beams and pollen tube growth.

Metabolites from Resistant and Susceptible <i>Pinus thunbergii</i>after Inoculation with Pine Wood Nematode

Pine wilt disease (PWD), which is caused by pine wood nematodes (PWN), is one of the most serious forest diseases worldwide. To clarify the mechanism of resistance to PWD, we compared metabolites from resistant and susceptible Japanese black pine (Pinus thunbergii) families after inoculation with PWN. After 2 weeks to 1 month post inoculation, the number of PWN dramatically increased in susceptible plants, but not in resistant plants. At this PWN-proliferation phase, ethyl acetate soluble fractions extracted from PWN-inoculated plants were analyzed by gas chromatogramphy-mass spectrometry (GC-MS). Although most compounds were qualitatively and quantitatively similar between resistant and susceptible plants, resistant plants accumulated 2.0-fold more linoleic acid (LA) than susceptible plants. On the other hand, benzoic acid (BA) was barely detected in resistant plants, but it accumulated in susceptible plants as the number of PWN increased. Susceptible plants contained greater levels of the nematicidal compounds pinosylvin and pinosylvin monomethyl ether, compared with resistant plants. These results suggested that LA is involved in the resistance reaction against PWN-proliferation, and that BA could be a good biomarker for PWD.

A Breakdown of Obligate Mutualism on a Small Island: An Interspecific Hybridization between Closely Related Fig Species (Ficus pumila and Ficus thunbergii) in Western Japan

Ficus (Moraceae) is a well-known group with specific pollination mutualisms, and hybridization is considered to be rare. Here, we report the presence of interspecific hybrids between Ficus pumila L. and F. thunbergii Maxim. on Okinoshima, a small island offshore of Shikoku, western Japan. AFLP (amplified fragment length polymorphism of genomic DNA) data suggested that more than one-fourth of individuals of morphological F. pumila were assigned as intermediate genotypes, suggesting hybrids. The hybridization between the two species was introgressive and unidirectional from F. thunbergii to F. pumila. The findings of this study, combined with other previous reports, suggest that the breakdown of mutualistic systems can occur in isolated populations such as those on islands.

Optimization of Ultrasonic Extraction Conditions of Polyphenols from the Needles of Black Pine (Pinus thunbergii) Using Response Surface Methodology (RSM)

Ultrasonic extraction conditions of polyphenols from the needles of black pine(Pinus thunbergii) were optimized by single factors analysis and Box-Behnken experiment design. The results showed that both of ethanol concentration and ultrasonic time had a significant effect on polyphenol extraction, and there was significant interaction between any two variables in the three parameters of ethanol concentration, ultrasonic temperature and ultrasonic time. The optimal parameters for polyphenol extraction were the ethanol concentration of 52.67%, ultrasonic temperature at 49.21 ℃and ultrasonic time of 30.76 min. Under these conditions, the actual content of polyphenols was close to the predicted value. Therefore, the process can be used to extract polyphenols from black pine needles.

鼠尾藻(Sargassum thunbergii)对邻苯二甲酸二甲酯(DMP)胁迫的响应

邻苯二甲酸二甲酯(dimethyl phthalate,DMP)是近海环境有机污染物之一,不仅对海洋生物造成毒害,且能在生物体内富集和放大,对海洋生态系统形成严重威胁。本文以鼠尾藻(Sargassum thunbergii)为研究对象,通过测定比生长速率、叶绿素a含量、光合和呼吸速率、光合基因rbcL转录水平上的相对表达量等指标,研究了鼠尾藻对不同浓度DMP的响应。结果表明,低浓度的DMP(0.lmg/L)短时间内(≤15d)能促进鼠尾藻的生长和叶绿素a的合成,并使藻体的光合作用增强,诱导光合基因rbcL的表达,而在高浓度DMP(≥0.3mg/L)的作用下,鼠尾藻的生长速率、光合和呼吸速率、rbcL基因的表达量均随着暴露时间的延长呈明显下降趋势,且DMP浓度越高,降幅越大。以上研究结果对了解DMP对鼠尾藻胁迫的影响机制奠定了基础,为保护海洋经济藻类的养殖环境提供了理论依据。

Comparative study of responses in the brown algae Sargassum thunbergii to zinc and cadmium stress

Heavy metal pollution in aquatic system is becoming a serious problem worldwide. In this study, responses of Sargassum thunbergii to dif ferent concentrations(0, 0.1, 0.5, 1.0 and 5.0 mg/L) of zinc(Zn) and cadmium(Cd) exposure separately were studied for 15 days in laboratory-controlled conditions. The results show that the specifi c growth rates increased slightly under the lower Zn concentration treatment(0.1 mg/L) at the first 5 d and then decreased gradually, which were significantly reduced with the exposure time in higher Zn concentrations and all Cd treatments compared to respective control, especially for 1.0 and 5.0 mg/L Cd. Chlorophyll a contents showed significant increase in 0.1 mg/L Zn treatment, whereas the gradually reduction were observed in the other three Zn treatments and all Cd treatments. The oxygen evolution rate and respiration rate presented distinct behavior in the Zn-treated samples, but both declined steadily with the exposure time in Cd treatments. The P/R value analyses showed similar variation patterns as chlorophyll a contents. Real-time PCR showed that lower Zn concentration(0.1 mg/L) increased mRNA expression of rbcL gene, whereas higher Zn concentrations and Cd reduced the rbcL expression. Taken together, these findings strongly indicate that Zn and Cd had different effects on S. thunbergii both at the physiological and gene transcription levels, the transcript level of photosynthesis-related gene rbcL can be used as an useful molecular marker of algal growth and environment impacts.

Performance of Weibull function as a diameter distribution model for Pinus thunbergii stands in the eastern coast of South Korea

This study was carried out to determine the performance of percentile-based Weibull diameter distribution model for Pinus thunbergii stands thriving along the eastern coast of South Korea. The parameter recovery technique was used to estimate the three parameters of the Weibull model. The analysis demonstrated satisfactory results based on the following test statistics for the principal percentile models： fit index （FI） range from 0.501（minimum diameter） to 0.932 （50th diameter percentiles） and root mean square error （RMSE） range from 0.112 （quadratic mean diameter） to 3.572 （minimum diameter）. The developed model was further evaluated by determining the mean bias （E） in trees per ha （TPH） for each diameter class, and the results showed highest over-prediction in the 20 cm, and under-prediction in the x6 cm and 24 cm diameter classes. The goodness of fit tested by Kolmogorov- Smirnov （KS） test showed no significant differences （P〉0.05） between the observed and predicted diameter distributions for almost all plots. Using site index and aboveground biomass （AGB） models developed for P. thunbergii in South Korea, a model to predict the AGB per ha for each diameter class and subsequently the total AGB of the stand was created. An application guide was also created, which will serve as a decision-support tool for forest managers in quantifying the future total AGB in P. thunbergii stands located in the eastern coast of South Korea and, subsequently, the quantification of potential carbon stocks aside from being a vital input in designing efficient management and protection strategies for these stands.

Modeling the Potential Distribution of <i>Machilus thunbergii</i>under the Climate Change Patterns in China

The potential geographic distribution and favorable climatic conditions of Machilus thunbergii under current and future predicted climates in China are predicted based on MaxEnt model and ArcGIS software. The results show that the AUC values in different time periods and emission seniors are more than 0.9, which indicates the prediction is excellent. Precipitation of the coldest quarter, precipitation of the driest month, annual precipitation, mean diurnal range, and temperature annual range are the most important environmental factors affecting the distribution of Machilus thunbergii. At present, the suitable areas of Machilus thunbergii are mainly concentrated in the eastern subtropics of China, with a total area of 118.47 × 104 km2. The medium-suitability area and the high-suitability area are concentrated in Wuyi Mountains, Luoxiao Mountains, Xuefeng Mountains, Nanling and east of Taiwan Mountains. With the change of climate, the suitable area increases, and the medium-suitability area and high-suitability area migrate and expand to the east, and the low-suitability area expands slightly to the west and north. So, the simulated distribution of Machilus thunbergii should be one of priorities, when instigating in-situ conservation. The research results can provide a theoretical reference for the popularization and planting of Machilus thunbergii.

Ion Implantation Hampers Pollen Tube Growth and Disrupts Actin Cytoskeleton Organization in Pollen Tubes of Pinus thunbergii

Pollen grains of Pinus thunbergii Parl. （Japanese black pine） were implanted with 30 keV nitrogen ion beams and the effects of nitrogen ion implantation on pollen tube growth in vitro and the organization of actin cytoskeleton in the pollen tube cell were investigated using a confocal laser scanning microscope after fluorescence labeling. Treatment with ion implantation significantly blocked pollen tube growth. Confocal microscopy showed that ion implantation disrupted actin filament cytoskeleton organization in the pollen tube. It was found that there was a distinct correlation between the inhibition of pollen tube growth and the disruption of actin cytoskeleton organization, indicating that an intact actin cytoskeleton is essential for continuous pollen tube elongation in Pinus thunbergii. Although the detailed mechanism for the ion-implantation-induced bioeffect still remains to be elucidated, the present study assumes that the cytoskeleton system in pollen grains may provide a key target in response to ion beam implantation and is involved in mediating certain subsequent cytological changes.

陌上菅(Carex thunbergii)对淹水弱光胁迫的形态生理响应

为研究陌上菅在淹水弱光条件下的响应机制,在室内模拟的水体中,采用人工遮荫的方法,设置了6个光强梯度,以株高、叶长、生物量、叶绿素a含量、抗氧化相关酶活性和丙二醛含量等为指标,探讨了淹水弱光环境对湿生植物陌上菅(Carex thunbergii)的形态生理影响。研究表明:(1)在持续淹水60d后,相对光强低至0.60%时陌上菅才开始出现死亡迹象,表明陌上菅具有很强的耐涝能力,在相对光强0.60%处理下,陌上菅仍能启动自身的抗性机制,表明相对光强0.60%左右水平可能是陌上菅淹水后受弱光环境胁迫的耐受下限;(2)淹水环境中,陌上菅抵御弱光胁迫的机制包括伸长叶长和株高、改变能量分配方式、提高叶绿素的含量和减缓新陈代谢等;(3)各弱光处理组MDA含量显著高于对照组,出现死亡迹象的处理组MDA含量高达对照组的40.17倍,同时抗氧化物酶活性与MDA含量呈一定的负相关。

Phenological study of Sargassum thunbergii (Fucales, Phaeophyta) in Lidao Bay, Rongcheng, China

We conducted a phenological evaluation of Sargassum thunbergii, a common algal species, along the coast of Lidao Bay, Rongcheng, China. The local S. thunbergii population entered a maturation period from early June to mid-August, when seawater temperature was 14.4 to 25. 1℃. Our results suggest an increase in temperatttre and day length during this period initiated thalli maturation. Inhabitants of the low tidal zone had higher thalli length and biomass compared with those from the mid- and high tidal zones. We observed a switch in the length frequency distribution and contribution of length classes to biomass between the small （〈20 cm） and long （20-79.9 cm） length classes at the time of thallus maturity. This suggests there is a trade-off between sexual reproduction and vegetative regeneration.

Impact of Environmental Factors on the Photosynthesis and Respiration of Young Seedlings of <i>Sargassum thunbergii</i>(Sargassaceae, Phaeophyta)

Sargassum thunbergii is of great economic and ecological value to sea cucumber cultures and seaweed beds. However, studies on photosynthesis and respiration of S. thunbergii are limited. In this study, a liquid-phase oxygen electrode system and a chlorophyll fluorescence spectrometer were used to determine the photosynthetic characteristics of S. thunbergii seedlings at various light intensities, temperatures, and salinities. The light-saturated net photosynthetic rates, light saturation points, and respiratory rates of germlings were investigated. Results showed that the increase in rate of light saturation point was slow in the first 4 d, rapidly increased from 5 d to 9 d, and then gently increased at the final few days of the 15 d indoor culture period. The photosynthetic rate or respiratory rate of the seedlings rapidly and significantly decreased when the temperature was °C or >28°C. Short-term high- or low-salinity shock had significant effect on the photosynthetic and respiratory rates of the seedlings, specifically at 10 and 50 psu. However, the photosynthetic and respiratory rates recovered to the normal levels after 24 h of recovery period, which demonstrated a powerful ion-transport system of the seedlings. These results provided reference for the artificial breeding of S. thunbergii.