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...     

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.     

蜂巢珊瑚类横板珊瑚的体壁构成及有关问题

一、前言本文所指蜂巢珊瑚类,包括所有具联接构造的块状复体横板珊瑚,即蜂巢珊瑚亚目(含蜂巢珊瑚超科、厚孔珊瑚超科)和槽珊瑚亚目(含槽珊瑚超科、小槽珊瑚超科)。多年来,在研究珊瑚体显微构造时,最先为人们注意的是体壁的构成。本文以“构成”一词代表构成体壁的部分,而以“微细构造”一词代表每一部分的构造细节。微细构造又分显微构造和超微构造两类,分别代表光学显微镜下和电子显微镜下所见的微细构造。由于光学显微镜放大倍数超过40倍后,视野昏暗,形象模糊,难以分辨。而横板珊瑚的基     

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S...     

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。     

深圳大鹏湾六种珊瑚共附生真菌多样性研究

本研究使用6种不同培养基对深圳大鹏湾6种珊瑚上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究深圳大鹏湾海域珊瑚共附生真菌的多样性。共分离培养出457株真菌菌株,分属于7个属,分别为红酵母属Rhodotorula（180株）、曲霉属Aspergillus（170株）、木霉属Trichoderma（50株）、青霉属Penicillium（34株）、枝孢属Cladosporium（21株）、透孢黑团壳属Massarina（1株）和弯颈霉属Tolypocladium（1株）,其中优势属为红酵母属Rhodotorula和曲霉属Aspergillus,占菌株分离总数的76.59%。单独鹿角珊瑚上分离到的共附生真菌数量和种类最多,为160株9种;在该种珊瑚上还分离出弯颈霉属Tolypocladium sp.,该属真菌此前从未在珊瑚共附生真菌的研究中分离出来。本研究还发现SDA培养基分离出的共附生真菌数量最多,CDA培养基的真菌数量最少,表明不同培养基分离的共附生真菌多样性存在差异。     

福建东山石珊瑚伴生物种多样性

本文记录了福建东山锯齿刺星珊瑚(Cyphastrea serrailia)、标准菊花珊瑚(Favia speciosa)和盾形陀螺珊瑚(Turbinaria peltata)等6种造礁石珊瑚。石珊瑚的立体生境中,栖息着埋栖、穴栖、缝栖、附着和游动等5种栖息习性的154种伴生物种。埋栖的连贵藤壶和穴栖的羽膜石蛏、珊瑚绒贻贝等是造礁石珊瑚伴生物种的表征。     

丛生盔形珊瑚共附生可培养真菌多样性分析

旨在研究珊瑚共附生可培养真菌多样性。运用稀释平板法和基于ITS-rDNA基因序列的系统发育分析对珊瑚共附生可培养真菌多样性进行研究, 将所得到的基因序列与NCBI数据库GenBank中的序列进行相似性比较并构建系统发育树。实验中从丛生盔形珊瑚表面和内部共分离得到19株菌落形态各异的真菌。ITS-rDNA序列分析及形态学鉴定表明, 丛生盔形珊瑚共附生真菌主要包括曲霉菌Aspergillus sp.、枝孢霉菌Cladosporium sp.、炭角菌Xylariales sp.、青霉菌Penicillium sp.、葡萄穗霉菌Stachybotrys sp.、赤霉菌Gibberella moniliformis、镰刀霉菌Fusarium sp.等。分离得到的菌株中, 4-13与GenBank中已报道的基因序列的相似性仅为89%。结果表明, 与丛生盔形珊瑚共附生的可培养真菌较为丰富, 是潜在的新的微生物菌种资源, 具有进一步研究的价值。     

鹿回头造礁石珊瑚群落多样性的现状及动态

通过对鹿回头岸礁（１０９°２８＇Ｅ,１８°１３＇Ｎ）造礁石珊瑚群落进行系统取样调查,分析了现有群落整体的成分组成,α－多样性和β－多样性及其在环境梯度上的变化;根据现有群落实际是处于不同演替时期地段的复合体的特点,把处于相同演替时期地段的综合特征作为分析群落整体在相应时期多样性特征的依据,探讨了群落演替过程中的多样性动态。研究发现：①中等水深（２．５￣５．０ｍ）地段的物种多样性平均值高于其他地段,     

鹿回头岸礁造礁石珊瑚物种多样性的研究

定量分析鹿回头岸礁造礁石珊瑚群落的种类组成,选择α－多样性和β－多样性的度量方法,分析了该群落造礁石珊瑚种、属和科的多样性及其与所处生境的关系。结果表明：鹿回头岸礁造礁石珊瑚群落的物种比较丰富,多样性和均匀度指数都较高,但在不同地段之间存在着明显的差异。水深中等的地段（２．４￣５．０ｍ）的多样性指数高于较浅（１．５￣２．０ｍ）和较深（５．５￣６．５ｍ）地段;潮间带与潮下带之间和礁平台与向海斜坡之间     

Diversity and dynamics of bacterial communities associated with Haematococcus pluvialis at different life stages

Journal of Applied Phycology - The physiological performance and productivity of microalgae are closely associated with their bacterial communities. Haematococcus pluvialis is an important...     

Patterns of association between Symbiodinium and members of the Montastraea annularis species complex on spatial scales ranging from within colonies to between geographic regions

Patterns of associations between coral colonies and the major clades of zooxanthellae can vary across scales ranging from individual colonies to widely separated geographic regions. This is exemplified in this study of the Montastraea annularis species complex from six sites on the Mesoamerican Reef, Belize and nine sites in the Bocas del Toro archipelago, Panama. Restriction fragment length polymorphism (RFLP) analysis of small subunit ribosomal DNA (SSU rDNA) was used to identify the zooxanthellae. In Belize (M. annularis), Symbiodinium B (79% of the colonies), Symbiodinium A, and Symbiodinium C were observed. In Panama (primarily M. franksi, but also M. annularis and M. faveolata), there was greater diversity and evenness with Symbiodinium A, B, C, C′ (a new symbiont) and D all being common in at least some host/habitat combinations. Non-metric multidimensional scaling ordinations showed that distribution patterns of symbionts across sites are best explained by enclosure (relative influence of open ocean vs. coastal water) and total suspended solids. Because members of clade D are known to be temperature resistant and Symbiodinium C′ was found in environments characterized by high sedimentation, these Panamanian reefs may have importance from a management perspective as reservoirs of corals better able to tolerate human impacts.     

Diversity of endophytic bacteria associated with nodules of two indigenous legumes at different altitudes of the Qilian Mountains in China

A total of 201 endophytic root nodule-associated bacteria collected from two legumes indigenous to different Qilian Mountain altitudes (Hexi Corridor) were characterized through 16S rDNA polymerase chain reaction (PCR)-restriction fragment length polymorphism, 16S rRNA gene sequence analysis, and enterobacterial repetitive intergenic consensus-PCR clustering. The isolates phylogenetically belonged to 35 species in the Phyllobacterium, Ensifer, Rhizobium, Microvirga, Sphingomonas, Paracoccus, Mycobacterium, Paenibacillus, Cohnella, Sporosarcina, Bacillus, Staphylococcus, Brevibacterium, Xenophilus, Erwinia, Leclercia, Acinetobacter, and Pseudomonas genera. Phylogenetic nodA sequence analysis showed higher similarity to Sinorhizobium meliloti with strains related to the Rhizobium, Sinorhizobium, and Acinetobacter genera. Sequence analysis of the nifH gene revealed that the strains belonging to Xenophilus, Acinetobacter, Phyllobacterium, and Rhizobium had genes similar to those of Mesorhizobium and Sinorhizobium. The results indicated that horizontal gene transfer could have occurred between rhizobia and non-rhizobial endophytes. Canonical correspondence analysis revealed that altitude and host plant species contributed more to the bacterial endosymbiont separation than other ecological factors. This study provided valuable information on the interactions between symbiotic bacteria, non-symbiotic bacteria and their habitats, and thus provided knowledge on their genetic diversity and ecology.     

岩溶型铝土矿尾矿堆不同自然演替阶段苔藓植物多样性特征

按空间代替时间序列的方法,对贵州省修文县小山坝岩溶型铝土矿区尾矿堆5个不同自然演替阶段苔藓植物多样性进行了研究。结果显示：（1）苔藓植物共有11科18属31种,其中藓类9科16属29种,苔类2科2属2种。在演替过程中,苔藓植物生活型单一,仅矮丛集型（占67.7%）和交织型（占32.3%）,矮丛集型占优势地位。苔藓植物物种组成简单,纯群落较多,混合群落随演替阶段逐渐增多。（2）不同自然演替阶段苔藓植物群落Shannon-Wiener指数、Pielou均匀度指数和丰富度指数有所不同,裸岩阶段最低（0.196、0.283和-1.930）,草灌丛阶段最高（3.470、1.281和2.342）,乔灌木阶段逐渐下降（2.128、0.887和1.267）。苔藓植物种的替代速率与相似性变化趋势相反。小山坝岩溶型铝土矿区尾矿堆苔藓植物多样性在5个不同自然演替阶段呈先上升后下降的变化规律,多样性水平总体呈上升趋势,研究结果可为岩溶型铝土矿区生态恢复和生物多样性保护提供基础资料。     

黄瓜初花期与结瓜期叶片可培养内生细菌多样性研究

【目的】研究黄瓜初花和结瓜两个生长期叶片可培养内生细菌的多样性。【方法】采用叶片表面消毒、菌种分离、16S rDNA序列扩增和系统发育分析进行了系统研究。【结果】两时期黄瓜内生细菌的种类、数量及优势菌的种类都有明显差异。初花期叶片含菌量为(2.6±0.18)106CFU/g鲜重,分离出的38株内生细菌分别属于短小杆菌属(Curtobacterium sp.)、节杆菌属(Arthrobacter sp.)、微杆菌属(Microbacterium sp.)等14个已知属,其中优势种类为短小杆菌属菌株;结瓜期叶片含菌量为(5.2±0.42)105CFU/g鲜重,分离出的43株内生菌分别属于泛菌属(Pantoea sp.)、假单胞菌属(Pseudomonas sp.)、芽孢杆菌属(Bacillus sp.)等11个已知属,其中优势种类为泛菌属菌株。【结论】初花期内生菌含量是结瓜期的5倍,两时期内生菌的种类表现出很强的差异,体现出黄瓜不同生育期可培养内生菌数量和种类的多样性,相关研究为黄瓜促生内生细菌的理论探索和生产应用提供一定的研究基础。     

Interaction of filipin with junctional membrane at different stages of the junction's life history

The relationship of filipin-sterol complexes to tight and gap junctions during their formation, maturation, internalization, and degradation was studied in separate cell lines. Filipin-sterol complexes tended to be excluded from mature junctions in tight junction forming COLO 316 cells and gap junction forming SW-13 cells. Once internalized, unlabeled junctional membrane appeared to fuse with heavily labeled vesicles, presumably lysosomes. Although the absence of filipin-sterol complexes from junctional membrane does not necessarily reflect the absolute sterol content of this membrane, the fact that filipin-sterol complexes are largely excluded from these areas indicates that this membrane is different from surrounding membrane. The absence of filipin-sterol complexes also permits the visualization of 'mixing' of this specialized unlabeled membrane domain with other filipin labeled membrane systems.     

根结线虫放线菌及其生物防治活性研究

从感染植物根部的根结线虫卵和雌虫中,分离得到放线菌20株。形态、细胞壁氨基酸组分和16S rRNA基因序列分析表明:分离菌株分属于链霉菌属(Streptomyces)、诺卡氏菌属(Nocardia)和假诺卡氏菌属(Pseudonocardia),其中链霉菌占80.0%。分离菌株对根结线虫Meloidogynespp.卵的平均寄生率、卵的孵化率、幼虫死亡率分别为54.1%、40.4%和26.2%。根据体外测试的结果,选择具有高致病力的3株链霉菌(1-17,2-6,9-47)和1株诺卡氏菌(5-1)进行温室番茄防效测试,其生防效率分别为31.4%、37.7%、56.4%和42.4%。     

Diversity of Vibrios associated with reared clams in Galicia (NW Spain)

The aim of the present study was to characterize and identify vibrios isolated from cultured clams in Galicia (NW Spain). A total of 759 isolates were obtained, phenotypically characterized, grouped and assigned to the genus Vibrio. Subsequently, the genomic diversity of 145 representative strains was analyzed by means of amplified fragment length polymorphism (AFLP), which revealed a high genetic diversity amongst these isolates. Only 57 out of 145 strains could be identified to the species level, and they were distributed in 13 AFLP clusters. V. cyclitrophicus, V. splendidus and V. alginolyticus were the most abundantly represented species. Eighty-eight isolates remained unidentified, 59 were distributed over 16 clusters, while 29 were unclustered. Sequencing of the 16S rRNA and two house-keeping genes (rpoA and recA) from representative strains belonging to eight unidentified clusters with the highest number of isolates confirmed their assignation to the Vibrionaceae family, and some of these probably represent new species within the genus. The present study confirmed that the phenotypic characterization of vibrios is not sufficient to identify them at the species level. A wide diversity of vibrios was found in cultured clams from all four geographic locations analyzed. In total, more than 12 Vibrio species and at least three potential new species in this genus were identified.