不同发育阶段的黄癣蜂巢珊瑚共生虫黄藻多样性

【背景】黄癣蜂巢珊瑚是我国南海主要的造礁石珊瑚之一,但至今未见对其共生虫黄藻的相关研究报道。【目的】研究黄癣蜂巢珊瑚不同发育阶段的共生虫黄藻多样性,探讨其获取途径。【方法】用荧光显微镜观察黄癣蜂巢珊瑚卵子、浮浪幼虫以及珊瑚成体中的共生虫黄藻。基于ITS2序列的克隆文库技术分析珊瑚卵子、浮浪幼虫、成体中共生虫黄藻的多样性。【结果】荧光显微镜观察结果显示,黄癣蜂巢卵子及发育至第4天的浮浪幼虫中均无共生虫黄藻。没有在黄癣蜂巢珊瑚卵子及4 d浮浪幼虫中检测到虫黄藻ITS2序列,但是在黄癣蜂巢珊瑚成体中检测到共生虫黄藻序列,系统发育分析提示为C1型虫黄藻。【结论】成体黄癣蜂巢珊瑚中发现了共生虫黄藻,而卵子、浮浪幼虫阶段均未发现虫黄藻,提示黄癣蜂巢珊瑚中的虫黄藻可能是后期从海水中水平转移而来,而不是由母体垂直遗传的。     

磷酸盐胁迫对造礁石珊瑚共生虫黄藻光合作用的影响

以佳丽鹿角珊瑚(Acropora pulchra)和多孔鹿角珊瑚(Acropora millepora)两种造礁石珊瑚为材料研究磷酸盐浓度对共生虫黄藻光合作用的影响.研究表明,佳丽鹿角珊瑚(A. Pulchra)和多孔鹿角珊瑚(A. Millepora)在对照组磷酸盐浓度下,共生虫黄藻的叶绿素荧光参数Fv/Fm值处于稳定状态,在15μmol/L和30μmol/L磷酸盐浓度下,虫黄藻的Fv/Fm值很快受到抑制,分别经过2d和5d开始慢慢恢复,但是最终只能恢复到比对照组较低的水平.同时两种珊瑚的共生虫黄藻密度也有所降低,尤其佳丽鹿角珊瑚(A. Pulchra)降低更加明显,15μmol/L和30μmol/L浓度下共生虫黄藻密度分别比对照组下降低5.59%和14.69%.因此, 佳丽鹿角珊瑚(A. Pulchra)和多孔鹿角珊瑚(A. Millepora)最大可以忍受30μmol/L的磷酸盐浓度,但是在30μmol/L浓度的耐受范围内,随着磷酸盐浓度的不断提高,珊瑚共生虫黄藻Fv/Fm值显著下降,珊瑚共生虫黄藻密度显著降低.     

珊瑚共生体中“细菌-虫黄藻-宿主”三角关系的通讯交流

“细菌-虫黄藻-珊瑚”是生态系统中一对经典的三角关系,其中包含着复杂的物质流、信息流和能量流,三者的平衡与稳定是维护珊瑚礁生态系统健康的重要保障。过去20年里针对共生体交互关系进行了大量研究,并取得了一些重要成果,明确了“细菌-虫黄藻-宿主”三者之间的物质代谢、营养交换以及与环境的交互关系。然而,基于共生系统的复杂性,一些现象背后的机制仍然未被充分揭示,尤其是共生体之间的通讯交流。信号分子介导的相互作用是珊瑚共生体稳态维持和高效运转的内在驱动力。本文以珊瑚共生体系中化学信号为重点,尝试梳理最新的研究进展,包括细菌与细菌、细菌与珊瑚、细菌与虫黄藻以及虫黄藻与珊瑚之间的通讯方式,重点关注了群体感应信号(QS)、二甲基巯基丙酸盐(DMSP)、糖类信号、脂类信号以及非编码RNA。选择性例举了QS信号介导的微生物协作和竞争、DMSP调节下的细菌和宿主的相互作用,以及环境胁迫下珊瑚和虫黄藻对非编码RNA的响应过程,强调了它们在共生体中的作用模式和生态意义。并对今后的研究重点和可能方向进行了提炼,包括研究维度的扩充、新技术-新方法的应用以及生态模型的构建等,旨在提升对三角关系互作方式的认识,增进对珊瑚共生体的理解,探索基于通讯语言的操纵方式为珊瑚礁生态系统的恢复和保护提供新思路。     

悬浮物对造礁石珊瑚营养方式的影响及其适应性研究进展

营养方式是造礁石珊瑚获取能量与营养物质的基础,影响其生长与分布。近年来珊瑚礁区悬浮物含量与组分结构发生显著变化,其对造礁石珊瑚营养方式的诸多影响正成为当前研究热点。研究系统综述了珊瑚礁区悬浮物变化特征、悬浮物对造礁石珊瑚营养方式的影响及其适应性研究现状。发现近年来人类活动加剧与强降雨事件频发是驱动珊瑚礁,尤其是近岸珊瑚礁区悬浮物含量递增、组分改变与变频加剧的主因;悬浮物变化对造礁石珊瑚光合自养与异养营养的影响存在显著的种间差异,这主要与悬浮物消光效应、生物可利用性及造礁石珊瑚种类密切相关。虽然少数种类造礁石珊瑚具光合可塑性或异养可塑性,能在高含量悬浮物存在的弱光环境中较好生长。然而对绝大多数造礁石珊瑚而言,其营养方式适应性较差,无法在悬浮物影响下较好地获取生命活动所需的能量与营养物质,进而难以生存。总体来说,悬浮物被认为是近年来影响造礁石珊瑚生长与分布的重要环境因子之一,而关于造礁石珊瑚营养方式对悬浮物变化的响应及其适应机制,当前研究仍较薄弱,需要进一步加强相关研究。     

珊瑚礁白化研究进展

珊瑚礁白化是由于珊瑚失去体内共生的虫黄藻和（或）共生的虫黄藻失去体内色素而导致五彩缤纷的珊瑚礁变白的生态现象。近年来，频繁发生的珊瑚礁白化导致了珊瑚礁生态系统严重退化，并已经影响到全球珊瑚礁生态系统的平衡，受到了人们的高度重视。研究认为：（1）大范围珊瑚礁白化主要是全球环境变化引起的，尤其是全球变暖和紫外辐射增强；（2）导致珊瑚礁白化的机制主要在于细胞机制和光抑制机制；（3）珊瑚礁白化后的恢复与白化程度有关，大范围白化的珊瑚礁完全恢复需要几年到几十年；（4）珊瑚礁白化的后果在于降低珊瑚繁殖能力、减缓珊瑚礁生长、改变礁栖生物的群落结构，导致大面积珊瑚死亡和改变珊瑚礁生态类型，如变为海藻型等；（5）与珊瑚共生的D系群虫黄藻更适应高温环境，珊瑚礁有可能通过D系群逐渐取代C系群的方式适应全球环境变化。     

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...     

虫黄藻在建造珊瑚礁中的作用

珊瑚虫是珊瑚礁的主要缔造者。各种不同类型的珊瑚虫都能分泌石灰质(CaCO_3) 外骨骼。由于个体大量繁殖,石灰质珊瑚骨越积越多,经过千百万年而形成珊瑚礁、珊瑚岛。然而不为一般人们所知的是:一种比珊瑚虫更小的虫黄藻在珊瑚虫建造珊瑚礁的过程中起着非同小可的作用。虫黄藻(Zooxanthella)是一种与珊瑚虫共生的单细胞植物。据估计,每 mm~3的珊瑚组织内有3万个虫黄藻,它们与珊瑚虫互惠共存。     

荧光标记珊瑚组织来源细菌及其与虫黄藻相互作用的显微观察

【背景】研究珊瑚-细菌、虫黄藻-细菌的相互作用是解析珊瑚健康机理的关键。对珊瑚共附生细菌进行稳定荧光标记有助于原位观察细菌与虫黄藻或珊瑚的相互作用。当前,对于野生型珊瑚共附生细菌遗传操作体系的研究有限,限制了对细菌与珊瑚、虫黄藻原位互作模式的揭示。【目的】建立一种适合专性海洋细菌的遗传操作体系,利用其对珊瑚组织来源细菌进行绿色荧光蛋白标记,用于研究标记菌株与虫黄藻的相互作用。【方法】通过电穿孔的方式将构建好的广宿主重组质粒转入供体菌(Escherichia coli WM3064),然后将供体菌与添加海水才可以生长的受体菌SCSIO 12696(港口球菌科,Porticoccaceae;分离自鹿角杯形珊瑚组织)按供、受体菌细胞数比分别为4:1、2:1、1:1比例混合,在25℃和30℃下于改良LB培养基上接合转移。显微观察标记细菌与虫黄藻相互作用。【结果】改良的LB培养基适用于需海水才可生长的专性海洋细菌的接合转移实验。接合转移的效率与供、受体菌的比例及温度有关。确定优化的接合转移条件为:供、受体菌的比例为1:1,温度为30℃。利用建立的接合转移体系,构建了增强型绿色荧光蛋白标记菌株SCSIO 12696。在激光扫描共聚焦显微镜下能清晰观察到标记菌株SCSIO 12696和虫黄藻在共培养时的相互作用。【结论】建立了适合专性海洋细菌的遗传操作体系,利用其构建荧光蛋白标记菌株可用于虫黄藻-细菌、珊瑚-细菌相互作用的研究,有助于揭示珊瑚共附生细菌的生态作用机制。     

卫星遥感珊瑚礁白化概述

珊瑚礁白化是由于珊瑚失去体内共生的虫黄藻或者共生的虫黄藻失去体内色素而导致五彩缤纷的珊瑚礁变白的现象,严重的白化可以带来珊瑚礁的死亡.国内外研究表明海水温度升高和珊瑚礁白化关系最为紧密.卫星遥感能够提供大范围、同步与连续的海洋数据,如海水表层温度和海色数据,从而能够及时监测和预测珊瑚礁的白化.基于AVHRR (Advanced Very High Resolution Radiometer),NOAA(National Oceanic and Atmospheric Administration,US)开发了全球监测珊瑚礁白化的方法,热点(HotSpot)和周热度(DHW)两种主要指数.目前,我国珊瑚礁白化现象的监测和研究明显滞后于国际动态,迫切需要发展和利用卫星遥感的方法监测南海珊瑚礁白化状况.     

甲藻的异养营养型

综述了甲藻的异养类型。目前已知异养营养型在甲藻中广泛存在,只有很少几种甲藻营严格自养营养方式。有近一半的甲藻物种是没有色素体的,还有很多甲藻即使具有色素体也会有异养营养需求,称为兼养营养类型。这些兼养类群不一定主要以有机物作为其获取碳的来源,而仅仅是补充一些生长必需的有机物如维生素、生物素等。兼养类群以渗透营养和腐食营养方式进行,同时也可以寄生方式和共生方式进行兼养生活。无色素体的甲藻以有机物作为碳的唯一来源,仅仅依靠异养方式生存,属于严格异养营养方式,又称有机营养型。它们是甲藻异养营养型的主体,其主要类型有寄生、渗透营养和吞噬营养。由于吞噬营养是甲藻异养的主要类型,因此论述了3种吞噬营养型:吞噬营养方式、捕食茎营养方式和捕食笼营养方式。吞噬营养方式在无甲类和具甲类甲藻中都有存在,主要通过甲藻细胞的纵沟或底部对猎物进行吞噬,也有研究发现吞噬部位为顶孔或片间带。捕食茎营养方式是通过捕食茎刺穿猎物细胞膜并吸食其细胞质来获取营养,在异养甲藻中也较常见。捕食笼营养方式只在原多甲藻属(Protoperidinium)和翼藻属(Diplopsalis)里发现,是甲藻通过鞭毛孔分泌细胞质到胞外形成捕食笼将猎物包裹并进行消化来摄食的。甲藻摄食对象尺寸范围变化较大,小至几微米,大至几百微米。有些甲藻具有摄食选择性,通过感应猎物释放的化学物质来判断猎物的位置并进行摄食,摄食完成后由于体积的增加经常会发生细胞分裂和蜕鞘。对于甲藻异养的其他形式如拦截摄食营养方式、伪足摄食营养方式、口足摄食营养方式、触手摄食营养方式等只作简单介绍。还就甲藻异养的研究方法、其生态学意义和进化学意义进行简要论述,并对相关研究进行展望。     

Heterotrophy in Tropical Scleractinian Corals

The dual character of corals, that they are both auto- and heterotrophs, was recognized early in the twentieth Century. It is generally accepted that the symbiotic association between corals and their endosymbiotic algae (called zooxanthellae) is fundamental to the development of coral reefs in oligotrophic tropical oceans because zooxanthellae transfer the major part of their photosynthates to the coral host (autotrophic nutrition). However, numerous studies have confirmed that many species of corals are also active heterotrophs, ingesting organisms ranging from bacteria to mesozooplankton. Heterotrophy accounts for between 0 and 66% of the fixed carbon incorporated into coral skeletons and can meet from 15 to 35% of daily metabolic requirements in healthy corals and up to 100% in bleached corals. Apart from this carbon input, feeding is likely to be important to most scleractinian corals, since nitrogen, phosphorus, and other nutrients that cannot be supplied from photosynthesis by the coral's symbiotic algae must come from zooplankton capture, particulate matter or dissolved compounds. A recent study showed that during bleaching events some coral species, by increasing their feeding rates, are able to maintain and restore energy reserves.
This review assesses the importance and effects of heterotrophy in tropical scleractinian corals. We first provide background information on the different food sources (from dissolved organic matter to meso- and macrozooplankton). We then consider the nutritional inputs of feeding. Finally, we review feeding effects on the different physiological parameters of corals (tissue composition, photosynthesis and skeletal growth).     

Heterotrophy mitigates the response of the temperate coral Oculina arbuscula to temperature stress

Anthropogenic increases in atmospheric carbon dioxide concentration have caused global average sea surface temperature (SST) to increase by approximately 0.11°C per decade between 1971 and 2010 – a trend that is projected to continue through the 21st century. A multitude of research studies have demonstrated that increased SSTs compromise the coral holobiont (cnidarian host and its symbiotic algae) by reducing both host calcification and symbiont density, among other variables. However, we still do not fully understand the role of heterotrophy in the response of the coral holobiont to elevated temperature, particularly for temperate corals. Here, we conducted a pair of independent experiments to investigate the influence of heterotrophy on the response of the temperate scleractinian coral Oculina arbuscula to thermal stress. Colonies of O. arbuscula from Radio Island, North Carolina, were exposed to four feeding treatments (zero, low, moderate, and high concentrations of newly hatched Artemia sp. nauplii) across two independent temperature experiments (average annual SST (20°C) and average summer temperature (28°C) for the interval 2005–2012) to quantify the effects of heterotrophy on coral skeletal growth and symbiont density. Results suggest that heterotrophy mitigated both reduced skeletal growth and decreased symbiont density observed for unfed corals reared at 28°C. This study highlights the importance of heterotrophy in maintaining coral holobiont fitness under thermal stress and has important implications for the interpretation of coral response to climate change.     

Coral bleaching is linked to the capacity of the animal host to supply essential metals to the symbionts

Massive coral bleaching events result in extensive coral loss throughout the world. These events are mainly caused by seawater warming, but are exacerbated by the subsequent decrease in nutrient availability in surface waters. It has therefore been shown that nitrogen, phosphorus or iron limitation contribute to the underlying conditions by which thermal stress induces coral bleaching. Generally, information on the trophic ecology of trace elements (micronutrients) in corals, and on how they modulate the coral response to thermal stress is lacking. Here, we demonstrate for the first time that heterotrophic feeding (i.e. the capture of zooplankton prey by the coral host) and thermal stress induce significant changes in micro element concentrations and isotopic signatures of the scleractinian coral Stylophora pistillata. The results obtained first reveal that coral symbionts are the major sink for the heterotrophically acquired micronutrients and accumulate manganese, magnesium and iron from the food. These metals are involved in photosynthesis and antioxidant protection. In addition, we show that fed corals can maintain high micronutrient concentrations in the host tissue during thermal stress and do not bleach, whereas unfed corals experience a significant decrease in copper, zinc, boron, calcium and magnesium in the host tissue and bleach. In addition, the significant increase in δ⁶⁵Cu and δ⁶⁶Zn signature of symbionts and host tissue at high temperature suggests that these isotopic compositions are good proxy for stress in corals. Overall, present findings highlight a new way in which coral heterotrophy and micronutrient availability contribute to coral resistance to global warming and bleaching.     

Oxygen and Heterotrophy Affect Calcification of the Scleractinian Coral Galaxea fascicularis

Heterotrophy is known to stimulate calcification of scleractinian corals, possibly through enhanced organic matrix synthesis and photosynthesis, and increased supply of metabolic DIC. In contrast to the positive long-term effects of heterotrophy, inhibition of calcification has been observed during feeding, which may be explained by a temporal oxygen limitation in coral tissue. To test this hypothesis, we measured the short-term effects of zooplankton feeding on light and dark calcification rates of the scleractinian coral Galaxea fascicularis (n = 4) at oxygen saturation levels ranging from 13 to 280%. Significant main and interactive effects of oxygen, heterotrophy and light on calcification rates were found (three-way factorial repeated measures ANOVA, p<0.05). Light and dark calcification rates of unfed corals were severely affected by hypoxia and hyperoxia, with optimal rates at 110% saturation. Light calcification rates of fed corals exhibited a similar trend, with highest rates at 150% saturation. In contrast, dark calcification rates of fed corals were close to zero under all oxygen saturations. We conclude that oxygen exerts a strong control over light and dark calcification rates of corals, and propose that in situ calcification rates are highly dynamic. Nevertheless, the inhibitory effect of heterotrophy on dark calcification appears to be oxygen-independent. We hypothesize that dark calcification is impaired during zooplankton feeding by a temporal decrease of the pH and aragonite saturation state of the calcifying medium, caused by increased respiration rates. This may invoke a transient reallocation of metabolic energy to soft tissue growth and organic matrix synthesis. These insights enhance our understanding of how oxygen and heterotrophy affect coral calcification, both in situ as well as in aquaculture.     

The importance of zooplankton to the daily metabolic carbon requirements of healthy and bleached corals at two depths

Bleached and non-bleached fragments of three species of Hawaiian corals were exposed to enhanced and ambient concentrations of zooplankton at 1 and 6 m depth to determine the contribution of zooplankton to the coral's daily carbon budget. The size and taxonomic grouping were recorded for every zooplankton captured and the relative input of zooplankton of different size classes was determined. The contribution of heterotrophy to animal respiration (CHAR) was calculated using an improved method that included the proportionate contribution of zooplankton from all size classes. Results show that the proportionate effects of species, depth and bleaching treatments on coral feeding rates were not significantly different between ambient and enhanced zooplankton concentrations. Corals captured the same size and assemblage of zooplankton under all evaluated conditions, and preferentially captured plankters smaller than 400 µm. Feeding rates of Porites lobata increased with depth regardless of bleaching status. Feeding rates of Porites compressa increased with depth in non-bleached corals, but not in bleached corals. Within depth, feeding rates of bleached Montipora capitata increased, P. compressa decreased and P. lobata remained unchanged relative to non-bleached fragments. Therefore, the feeding response of corals to the same disturbance may vary considerably. Calculated CHAR values show that heterotrophic carbon from zooplankton plays a much larger role in the daily carbon budget of corals than previously estimated, accounting for 46% of some coral species' daily metabolic carbon requirements when healthy and 147% when bleached. Thus, heterotrophically acquired carbon made an important contribution to the daily carbon budget of corals under all experimental conditions. These results suggest that the relative importance of autotrophic and heterotrophic carbon to a coral's energetic needs is mediated by a coral's bleaching status and environment, and should be considered on a continuum, from 100% photoautotrophy to 100% heterotrophy.     

Calcification by juvenile corals under heterotrophy and elevated CO2

Ocean acidification (OA) threatens the existence of coral reefs by slowing the rate of calcium carbonate (CaCO₃) production of framework-building corals thus reducing the amount of CaCO₃ the reef can produce to counteract natural dissolution. Some evidence exists to suggest that elevated levels of dissolved inorganic nutrients can reduce the impact of OA on coral calcification. Here, we investigated the potential for enhanced energetic status of juvenile corals, achieved via heterotrophic feeding, to modulate the negative impact of OA on calcification. Larvae of the common Atlantic golf ball coral, Favia fragum, were collected and reared for 3 weeks under ambient (421 μatm) or significantly elevated (1,311 μatm) CO₂ conditions. The metamorphosed, zooxanthellate spat were either fed brine shrimp (i.e., received nutrition from photosynthesis plus heterotrophy) or not fed (i.e., primarily autotrophic). Regardless of CO₂ condition, the skeletons of fed corals exhibited accelerated development of septal cycles and were larger than those of unfed corals. At each CO₂ level, fed corals accreted more CaCO₃ than unfed corals, and fed corals reared under 1,311 μatm CO₂ accreted as much CaCO₃ as unfed corals reared under ambient CO₂. However, feeding did not alter the sensitivity of calcification to increased CO₂; ? calcification/?Ω was comparable for fed and unfed corals. Our results suggest that calcification rates of nutritionally replete juvenile corals will decline as OA intensifies over the course of this century. Critically, however, such corals could maintain higher rates of skeletal growth and CaCO₃ production under OA than those in nutritionally limited environments.     

Nitrogen and photosynthetic function of hermatypic corals. Oxygen exchange of Stylophora pistillata coral under artificial feeding

The change of Stylophora pistillata coral photosynthetic function (oxygen exchange and biomass of symbionts) under starvation and food enrichment was studied to understand the role of heterotrophy in nitrogen supplements of zooxanthellae. The starvation caused the decrease of frequency of zooxanthellae cells division in 7-10 times. The number of degraded algae cells increased in same proportion and, as a result, the density of zooxanthellae in corals decreased about two times during one-two weeks. Under starvation corals kept their photosynthetic capacity at the level of corals in situ by means of enhancing the zooxanthellae gross photosynthesis. The respiration rate of coral had tendency to increase and the dry mass of polyp tissue to decrease. Under artificial feeding which was following starvation the zooxanthellae density increased in 1.5-2 times, and particular food caused more intensive accumulation of zooxanthellae comparing to dissolved inorganic ammonium. The feeding regime did not affect dry mass of polyp tissue and chlorophyll content as well as respiration and gross productivity of the corals. The conclusion about high effectiveness of particular feeding for supplying symbiotic algae with nitrogen was made and trophic status of zooxanthellae in hospite was determined as unlimited by nitrogen.