造礁石珊瑚共生虫黄藻离体培养方法的优化

【目的】开发一种高效地从造礁石珊瑚中分离、培养共生虫黄藻的技术方法,为珊瑚共生虫黄藻藻种资源储备和生理功能研究积累基础。【方法】首先采用微孔滤网过滤法和密度梯度离心法从造礁石珊瑚组织中直接分离或富集共生虫黄藻细胞,然后用改良的L1培养基在96孔板上对所得细胞进行离体培养,最后进行单细胞分离、培养和(或)平板划线培养获得单克隆虫黄藻细胞系。对所得虫黄藻单克隆藻株进行聚合酶链式反应-限制性内切酶片段长度多态性(polymerase chainreaction-restrictionfragmentlengthpolymorphism,PCR-RFLP)分析,结合内转录间隔区2(internal transcribed spacer2,ITS2)和大亚基(large subunit,LSU)测序进行物种鉴定及系统发育分析。【结果】采用上述方法从涠洲岛的霜鹿角珊瑚(Acropora pruinose)和西沙群岛的丛生盔形珊瑚(Galaxea fascicularis)及柔枝鹿角珊瑚(Acropora tenuis)中分离、培养得到3个虫黄藻株系,编号分别为AP21C1、GF21D1和AT21A...     

Effects of the multiple stressors high temperature and reduced salinity on the photosynthesis of the hermatypic coral Galaxea fascicularis

This study investigates the physiological responses in the hermatypic coral Galaxea fascicularis exposed to salinity stress (from 37 ppt to 15 ppt) for 12 h, combined effects of reduced salinity (from 37 ppt to 20 ppt) and two temperatures (26 °C and 32 °C) for 12 h and combined effects of reduced salinity (from 37 ppt to 25 ppt) and two temperatures (26 °C and 29.5 °C) for 10 d. The results demonstrate that the coral is tolerant to 12 h exposure to extremely low salinity (15 ppt). The study also shows that combined effects of temperature and low salinity aggravate the damage on the photosynthesis of the symbiotic dinoflagellates in 12 h exposure to 20 ppt sea water. This study suggests that high temperature (29.5 °C) aggravates the damage of trivially low salinity (30 ppt) on the holobiont (the coral and its symbiotic dinoflagellates) in 10 d exposure. However, high temperature (29.5 °C) may have an antagonistic effect between temperature and low salinity (25 ppt) on metabolism of the holobiont. Based on the above results, we suggest that (1) the true mechanism of corals exposed to combined effects of low salinity and high temperature is complicated. This calls for more studies on different corals. Future studies should aim at investigating long-term low-level stress in order to simulate in situ conditions more accurately; (2) when corals exposed to extremely severe combined stressors for short-term or trivially severe stressors for relative long-term, the combined effects of two stressors (such as low salinity and high temperature) may be negative, otherwise, the effects may be additive.     

珊瑚及共生藻在白化过程中的适应机制研究进展

珊瑚礁生态系统具有非常重要的生态学功能。但是随着全球气候变暖和CO2浓度的升高,珊瑚白化事件越来越频繁,珊瑚礁生态系统面临严重的危机。影响珊瑚白化的重要因子主要有海水温度的异常（过高或过低）,太阳辐射与紫外线辐射,海水盐度的偏离,珊瑚疾病,海洋污染,长棘海星的爆发,人类的过度捕鱼和全球CO2浓度升高等。其中,海洋表面水体温度（SST）的异常升高为珊瑚白化的主要因素。珊瑚主要是通过珊瑚与共生藻的生理适应机制以及更换共生藻基因型机制两种方式来适应环境胁迫的。生理适应机制主要通过叶黄素循环、珊瑚色素荧光（热）、活性氧清除系统（自由基）、分泌紫外线吸收物质MAAs（紫外光）、产生热休克蛋白HspS（热）来实现的。珊瑚共生藻基因型更换适应机制是指珊瑚的适应性白化假说。珊瑚的适应性白化假说还有很多争议,还需要更多的实验证据提供支持。未来的研究重点将在珊瑚白化过程中共生藻-珊瑚共生功能体作为整体性的研究,尤其是珊瑚宿主在白化过程中对共生功能体作出贡献的研究。     

Diversity of symbiotic algae of the genus Symbiodinium in scleractinian corals of the Xisha Islands in the South China Sea

Symbiotic algae （Symbiodinium sp.） in scleractinian corals are important in understanding how coral reefs will respond to global climate change. The present paper reports on the diversity of Symbiodinium sp. in 48 scleractinian coral species from 25 genera and 10 families sampled from the Xisha Islands in the South China Sea, which were identified with the use of restriction fragment length polymorphism （RFLP） of the nuclear ribosomal DNA large subunit gene （rDNA）. The results showed that： （i） Symbiodinium Clade C was the dominant zooxanthellae in scleractinian corals in the Xisha Islands; （ii） Symbiodinium Clade D was found in the corals Montipora aequituberculata, Galaxea fascicularis, and Plerogyra sinuosa; and （iii） both Symbiodinium Clades C and D were found simultaneously in Montipora digitata, Psammocora contigua, and Galaxeafascicularis. A poor capacity for symbiosis polymorphism, as uncovered by RFLP, in the Xisha Islands indicates that the scleractinian corals have low adaptability to environmental changes. Further studies are needed to investigate zooxanthellae diversity using other molecular markers.     

Effects of winter cold stress on photosynthesis and photochemical efficiency of PSII of the Mediterranean <Emphasis Type="Italic">Cistus albidus</Emphasis> L. and <Emphasis Type="Italic">Quercus ilex</Emphasis> L.

This study examined the photosynthetic and growth performances of potted plants of Cistus albidus L. and Quercus ilex L. submitted either to natural Mediterranean winter conditions or to mild greenhouse conditions. Plants grown outdoors exhibited lower light and CO₂-saturated CO₂ assimilation rates (A_sat) and apparent quantum yield (_i) than those indoors. Until mid-winter, C. albidus had higher A_sat than Q. ilex, but differences disappeared after a period of severe cold. Maximal photochemical efficiency of PSII (Fv/Fm) measured predawn was higher in C. albidus than in Q. ilex, and decreased throughout the season in outdoor plants. Fv/Fm also decreased at light saturation (A_sat) in both species. Fv/Fm was correlated with photosynthetic capacity and efficiency (quantum yield), but the resulting regression slopes were different between the two species. At the physiological level, C. albidus seemed to cope better with cold stress than Q. ilex. However, winter stress induced reduction of leaf absorptance, increased leaf mass per area, extensive leaf damage and high plant mortality in C. albidus. This suggests that the high performance of C. albidus leaves is not likely to be maintained for long periods of cold stress, and may therefore depend on continuous leaf replacement. Quercus ilex showed a conservative behaviour, with low net assimilation rates but greater leaf and plant survival than C. albidus.     

干旱对白皮松幼苗针叶生理特性的影响

试验目的是研究栽植前和栽植后干旱致使白皮松（Pinus bungeana Zucc.）针叶的生理特性的改变,了解白皮松造林前后能够适应的干旱范围,为白皮松科学造林提供理论依据。试验中模拟白皮松容器苗造林前后可能遇到的干旱胁迫,研究栽植前容器苗分别为B1：75%～80%（正常浇水）、B2：55%～60%（轻度干旱）、B3：35%～40%（严重干旱）和栽植后土壤相对含水量分别为A1：75%～80%、A2：55%～60%、A3：35%～40%、A4：15%～20%（极严重干旱）对白皮松针叶的生理指标的影响。结果表明：栽植前白皮松容器苗经B3处理后,针叶电解质渗透率增加,叶绿素含量和PSII最大光化学效率（Fv/Fm）显著降低,表明针叶受到一定伤害。栽植后如果正常浇水,这些影响将消除,如果遭遇栽植后干旱,B3处理的受害程度不比B1和B2处理深。栽植后白皮松经A3和A4处理4周后,针叶的电解质渗透率增加,Fv/Fm降低,表明白皮松针叶受到伤害。白皮松容器苗造林在栽植前遭到适当干旱胁迫时不会影响造林后的表现。但应避免栽植后初期4周以上严重干旱胁迫。     

不同培养基对兰科药用植物手参原球茎共生真菌的分离效果

兰科植物手参Gymnadenia conopsea作为国家二级重点保护野生植物具有重要的药用价值。目前手参还未实现人工栽培,但其种子的真菌共生萌发已获成功。为明确除促萌发真菌外,还有哪些土著真菌参与了手参种子的萌发过程,本研究在自然条件下采用促萌发真菌伴播手参种子,获得了种子萌发形成的原球茎,进而对比了6种常见的培养基PDA (马铃薯葡萄糖琼脂培养基)、MMN (改良Melin-Norkrans培养基)、FIM (真菌分离培养基)、MEA (麦芽浸膏琼脂培养基)、CAM (胡萝卜葡萄糖琼脂培养基)和CMA (玉米粉琼脂培养基)对手参原球茎共生真菌分离效果的影响。共从6种培养基上分离获得了75个菌株,其中MMN、CAM、PDA、FIM、MEA、CMA培养基依次分离得到20株、16株、15株、11株、8株、5株菌。此外,真菌的多样性分析结果表明,MMN培养基的Chao 1、Shannon-Wiener和Simpson多样性指数最高,CAM和PDA培养基次之,CMA培养基最低。综上所述,真菌分离效果最好的是MMN培养基,其次是CAM和PDA培养基,而FIM和MEA培养基对真菌的分离效果影响不大,CMA培养基的分离效果最差。本研究结果可为其他兰科植物原球茎共生真菌的分离提供借鉴,所获得的菌株也有望进一步应用于功能菌剂的研发。     

Rhythmical changes in the division and degradation of symbiotic algae in hermatypic corals

Three types of rhythmical changes in division and degradation (digestion) have been found and described in the symbiotic zooxanthellae of the scleractinian coral Seriatopora caliendrum and in the colonial hydroid Myllepora intricata collected at the reef of Sesoko Island (Okinawa, Japan). The first type of changes reported for S. caliendrum is daily variation of the intensity of these two processes peaking at night. The second type of rhythm changes described in S. caliendrum and M. intricata is opposing variations of cell division and degradation levels with a period of three days. The rhythm is disturbed by sudden weather changes or by variations in light. Rhythmical alterations of zooxanthella proliferation and degradation of the third type found in S. caliendrum have a period of 5–6 days and are connected with oppositely directed changes as well as with alterations of the second type. The rhythm of such alterations is not disturbed by abrupt weather changes. All three types of rhythmical changes in zooxanthellae division and digestion observed in hermatypic coral are supposed to be directed to the regulation of the population density of zooxanthellae in organism tissues.     

Variation in responses to temperature treatments ex situ of the moss Pleurozium schreberi (Willd. ex Brid.) Mitt. originating from eight altitude sites in Hokkaido,Japan

Thermal acclimatisations are important for the survival and growth of individuals and populations but seldom studied for different populations of bryophytes. The aims of this study were to (I) investigate if responses to temperature treatments were independent of the site sampled or if the intra- and inter-population variation in responses were larger than the responses to the temperature treatments (control, press, and pulse), and to (II) examine if experimental responses varied, depending on the sampled sites. We collected samples of the circumpolar bryophyte species, Pleurozium schreberi (Willd. ex Brid.) Mitt., originating from eight altitude sites on Mt. Oakan in Hokkaido, Japan, and exposed them to three different temperature treatments ex situ for four weeks. Thermal acclimatisation was estimated by measuring responses in growth length increase, biomass increase, number of branches, and the maximum quantum yield of PS II (Fv/Fm). We found that responses to temperature treatments were dependent on the site sampled, and that differences were most pronounced in the length increase. Results also shows that the responses to experimental treatments may differ between sites. Our results therefore raise important concerns regarding the general validity of both ex situ and in situ experiments when performed on a single or a limited number of sites.     

光照和营养盐磷对微型及微微型浮游植物生长的影响

2004年9月,在长江口及邻近水域通过在培养水体中添加不同量的磷酸盐和改变光照强度进行现场受控培养实验,对光照和营养盐磷耦合培养作用下浮游植物生长及对磷营养盐的吸收变化进行了研究,结果表明：高光照条件下（100﹪自然光照）,磷酸盐浓度在高磷水平（0.60μmol/L）培养水体中下降速率明显比中磷（0.41μmol/L）、低磷水平（0.25μmol/L）快,浮游植物生长存在着显著的磷限制性,微型浮游植物（nanophytoplankton,简称Nano,2～20μm）在高磷水平下的生长明显得到促进,聚球藻（Synechococcus sp.,简称Syn,＜2μm）密度在培养初期有小幅度增加,而微微型真核浮游植物（picoeukaryote,简称Euk,＜2μm）在低磷水平下生长较快;在低光照条件下（50﹪自然光照）,磷酸盐浓度在高磷水平培养水体中的下降是受到抑制的,Nano和Syn也都更宜在中磷水平培养水体中生长,Euk在高磷水平下的生长也是受到抑制的,且在中磷水平培养水体中,三类浮游植物的生长周期都得到延长;无光照暗环境培养条件下磷酸盐浓度在不同磷水平下始终保持着增加趋势,三类浮游植物也都无法生长,磷酸盐浓度随培养时间呈线性增加趋势,浮游植物细胞密度则呈指数下降趋势,且磷酸盐的添加对其本身的释放速率和浮游植物衰减速率都没有影响.     

Effects of di-n-butyl phthalate on growth and photosynthesis in algae and on isolated organelles from higher plants

Di- n -butyl phthalate (DBF) is widely used as a plasticizer and has been found in all types of ecosystems. It inhibits growth and photosynthesis of green algae ( Chlorella emersonii CCAP strain 211/8 h and Selenastrum capricornutum CCAP strain 278/4) at concentrations higher than 10-⁵ M . The IC₅₀ value for CO₂-dependent oxygen evolution in algae was 3 × 10^-4M. The CO₂-reduction in isolated protoplasts prepared from barley ( Hordeum vulgare L. cv. Simba) was also inhibited by phthalate. The IC₅₀ value was 2 × 10^-4 M . The electron transport in isolated thylakoids prepared from spinach was inhibited with an IC₅₀ value of 3 × 10^-4 M . The IC₅₀ value for uncoupled electron transport extrapolated to zero chlorophyll concentration was 2.5 × 10^-5 M . The effect of di-n-butyl phthalate was localized to reactions in photosystem II. Di-n-butyl phthalate could thus be a pollutant which affects growth and photosynthesis of plants. The reported IC₅₀ values may be underestimated since di- n -butyl phthalate can attach to surfaces. The results are discussed in relation to observed effects of di- n -butyl phthalate on other organisms.