温度、盐度对马氏珠母贝鳃Hsp70基因表达量的联合效应

采用中心复合设计和响应曲面分析法,在实验室条件下研究了温度(16 ～40℃)、盐度(10 ～50)对马氏珠母贝鳃热休克蛋白Hsp70基因表达量的联合效应.设定温度范围为16～40℃,盐度范围为10～50.结果表明:温度的一次效应、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著;盐度的一次效应对马氏珠母贝鳃Hsp70基因表达量无显著影响、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著,温度、盐度之间的互作效应对马氏珠母贝鳃Hsp70基因表达量无显著影响,且温度的效应大于盐度.建立了马氏珠母贝鳃Hsp70基因表达量模型方程,决定系数98.7％、矫正决定系数97.4％,预测决定系数89.2％,模型的拟合度极高,可用于预测马氏珠母贝鳃Hsp70基因的表达量.通过对模型方程优化,得到在温度26.78℃、盐度29.33时,马氏珠母贝鳃Hsp70基因表达量达到最小值0.5276,满意度达到98％.试验结果可为马氏珠母贝鳃Hsp70基因的上调表达、维持细胞内环境稳定以及提高马氏珠母贝抗逆性提供理论指导.     

温度、盐度对马氏珠母贝鳃Hsp70基因表达量的联合效应

采用中心复合设计和响应曲面分析法,在实验室条件下研究了温度(16～40 ℃)、盐度(10～50)对马氏珠母贝鳃热休克蛋白Hsp70基因表达量的联合效应.设定温度范围为16～40 ℃,盐度范围为10～50.结果表明: 温度的一次效应、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著;盐度的一次效应对马氏珠母贝鳃Hsp70基因表达量无显著影响、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著,温度、盐度之间的互作效应对马氏珠母贝鳃Hsp70基因表达量无显著影响,且温度的效应大于盐度.建立了马氏珠母贝鳃Hsp70基因表达量模型方程,决定系数98.7%、矫正决定系数97.4%,预测决定系数89.2%,模型的拟合度极高,可用于预测马氏珠母贝鳃Hsp70基因的表达量.通过对模型方程优化,得到在温度26.78 ℃、盐度29.33时,马氏珠母贝鳃Hsp70基因表达量达到最小值0.5276,满意度达到98%.试验结果可为马氏珠母贝鳃Hsp70基因的上调表达、维持细胞内环境稳定以及提高马氏珠母贝抗逆性提供理论指导.     

沉积再悬浮颗粒物对马氏珠母贝摄食生理影响的室内模拟

采用实验生态学方法室内模拟研究了不同浓度沉积再悬浮颗粒物对马氏珠母贝清滤率、摄食率、吸收率的影响。结果表明:(1)水体中总悬浮颗粒物对马氏珠母贝清滤率的影响极显著(P<0.01)。总悬浮颗粒物由低浓度(12.6 mg/L)趋高浓度(500 mg/L)时,马氏珠母贝的清滤率呈现峰值变化规律。与总悬浮颗粒物浓度50 mg/L时的最大清滤率(1.12 L.个体-1.h-1)比较,悬浮颗粒物浓度为500 mg/L时,清滤率达最小值(0.17 L.个体-1.h-1)),其清滤率降幅达85%。这表明在高浓度悬浮颗粒物的水环境下,贝类受到环境胁迫,其生理和自身摄食机制受到限制,引起摄食减少和机体损伤。马氏珠母贝类的清滤率(CR)与总悬浮颗粒物浓度(TPM)之间的关系可表达为:CR=-0.701+1.627×TPM-0.463×TPM2+0.036×TPM3(R2=0.928)。(2)水体中总悬浮颗粒物对马氏珠母贝摄食率的影响极显著(P<0.01)。马氏珠母贝的摄食率随着总悬浮颗粒物浓度的升高而增加,在50 mg/L时达最大值(38.28 mg/h),当总悬浮颗粒物浓度超过50 mg/L时,摄食率反而下降,在总悬浮颗粒物浓度为500 mg/L时,降为最小值(16.22 mg/h),摄食率降幅为58%。随着悬浮颗粒物浓度的增加,马氏珠母贝摄食率受到的影响小于清滤率受到的影响。马氏珠母贝类的摄食率(IR)与总悬浮颗粒物浓度(TPM)之间的关系可表达为:IR=-46.631+70.957×TPM-18.385×TPM2+1.367×TPM3(R2=0.907)。(3)水体中总悬浮颗粒物对马氏珠母贝吸收率影响极显著(P<0.01)。总悬浮颗粒物由低浓度(12.6 mg/L)趋高浓度(500 mg/L)时,马氏珠母贝的吸收率呈逐渐下降趋势,在总悬浮颗粒物12.6 mg/L时,马氏珠母贝的吸收率最大(48.57%),而总悬浮颗粒物500 mg/L时,马氏珠母贝的吸收率最小(8.56%)。马氏珠母贝的吸收率(AE)与总悬浮颗粒物浓度(TPM)之间的关系可表达为:AE=52.189+0.132×TPM-3.111×TPM2+0.316×TPM3(R2=0.976)。     

缺氧与饥饿对马氏珠母贝滤水率和耗氧率的影响

为了揭示缺氧和饥饿对马氏珠母贝的生理能量学的影响,设置对照组、缺氧组、饥饿组、缺氧且饥饿组,处理12 h后连续5 d分析滤水率与耗氧率,结果显示缺氧和饥饿都显著影响马氏珠母贝的滤水率(p0.05),但是缺氧和饥饿对马氏珠母贝的滤水率不存在显著交互作用(p0.05);无论是缺氧还是饥饿均对对马氏珠母贝的耗氧率没有显著影响(p0.05),而且缺氧和饥饿对马氏珠母贝的耗氧率也没有交互作用(p0.05)。     

香港巨牡蛎和长牡蛎幼虫及稚贝的表型性状

为了评估香港巨牡蛎和长牡蛎在北方沿海的早期表型性状,于2010年7月,以2009年6月在青岛繁育的两种牡蛎为材料,在大连研究了温度(Mt:(22±1.0)℃及Ht:(28±1.0)℃)、盐度(S₂₀:20±1.0及S₃₀:30±1.0)及中间育成环境(ID:室内及OD:室外)对两种牡蛎幼虫及稚贝表型性状的影响。结果表明:香港巨牡蛎壳宽显著大于长牡蛎(P<0.05),壳高及怀卵量显著小于长牡蛎(P<0.05),壳长、鲜重及壳重两者间无显著差异(P>0.05)。在温度和盐度相同情况下,长牡蛎卵径、受精率、孵化率及D形幼虫均大于香港巨牡蛎;香港巨牡蛎幼虫浮游前期生长较慢,而后快于长牡蛎。两种牡蛎幼虫存活能力在15日龄时高温组>中温组;相同温度下,香港巨牡蛎中盐组>高盐组,长牡蛎高盐组>中盐组。幼虫变态期间,较低的温度延迟了变态时间,降低了变态率,使得两种幼虫变态规格大型化。温度是影响幼虫生长、存活、变态的最主要因素,其次为盐度,交互作用几乎尚未起到作用。中间育成阶段,室外比室内培育效果更好,且香港巨牡蛎稚贝壳高在60日龄以后显著大于长牡蛎(P<0.05),环境是影响稚贝生长的最主要因素;无论室内还是室外两种牡蛎稚贝的存活率均在90%以上,且各实验组间无显著差异(P>0.05)。     

合浦珠母贝鳃的显微与超微结构

合浦珠母贝(Pinctada fucata)是典型的滤食性瓣鳃类动物,也是我国重要的海水珍珠养殖贝类。本研究用光学显微镜、扫描电镜和透射电镜观察了合浦珠母贝鳃的显微和超微结构。结果表明,合浦珠母贝鳃结构属于异丝鳃型,左右两侧各2个鳃瓣,每个鳃瓣由内鳃瓣和外鳃瓣组成。鳃瓣由主鳃丝和普通鳃丝构成,主鳃丝在鳃瓣中主要起支架作用,每2根主鳃丝之间的9～12根普通鳃丝由"簇内连接"(intrabunchial junction)相连成簇。普通鳃丝之间通过"丝间连接"(interfilament junction)相连,丝间连接的上皮细胞与普通鳃丝的扁平细胞结构一样,为鳃的呼吸上皮。丝间连接的存在扩大了鳃的表面积,这种结构有助于进行气体交换。主鳃丝和普通鳃丝表面有前纤毛和侧纤毛,与食物运送和气体交换有关。普通鳃丝表面的纤毛为典型的"9+2"型微管结构。     

不同盐度、pH条件下氨氮对管角螺稚贝毒性影响

在水温28.5℃,采用实验生态学方法研究了不同盐度、pH条件下氨氮对管角螺(Hemifusus tuba)稚贝[壳高(11.3±0.11)mm,n=30]毒性的影响。结果表明,(1)盐度对氨氮的毒性有较大影响,随着盐度的降低,氨氮在水体中的毒性增强;盐度为16、19、23和28,总氨氮对稚贝的96h半数致死浓度(96hLC50)分别为36.5、43.7、52.6和58.8mg/L,安全浓度(SC)为3.7、4.4、5.3和5.9mg/L。(2)氨氮在水体中的毒性随pH的升高而增强,pH为7.6、8.0、8.4和8.8时,总氨氮对稚贝的96hLC50依次为58.3、54.5、50.6和20.2mg/L,对应的SC依次为5.8、5.5、5.1和2.0mg/L。氨氮在pH8.8时对稚贝的96hLC50急剧下降,其毒性是在pH7.6时的2.9倍。     

温度和盐度对青蛤孵化及幼虫、稚贝存活与生长变态的影响

在9个温度梯度(10-34℃)和10个盐度梯度(盐度3‰-50‰)条件下,研究了温度和盐度对青蛤孵化及幼 虫、稚贝生存与生长变态的影响。结果表明,青蛤孵化和浮游幼虫生长的适温范围为24-32℃,最适温度均为26- 30℃,稚贝生长的适温范围为22-32℃,最适温度为24-30℃。在最适温度下,D形幼虫变态率达80．7％-88．2％, 浮游幼虫和稚贝的存活率分别为86．2％-88．7％和81．5％-84．0％;孵化及浮游幼虫的生长适宜盐度为15‰- 30‰,稚贝为10‰-35‰,最适盐度均为20‰-25‰。在最适盐度下,D形幼虫的成活率、变态率、生长速度皆最高, 分别达到86．9％、77．5％和9．38×11．0μm/d,匍匐幼虫经14-14．5d发育至双管期稚贝,至双管期稚贝的成活率 82．5％-85．0％,日平均生长达13．1μm以上。与大多数滩涂贝类一样,青蛤属于广温广盐性贝类,且稚贝对低盐的 适应能力强于对高盐的适应能力。     

马氏珠母贝Sox11基因的克隆及时序表达模式分析

为探究Sox(SRY-related HMG-box genes)基因家族在马氏珠母贝个体发育及性别分化中的作用, 研究首先利用兼并引物从马氏珠母贝基因组中克隆到一个HMG框(high mobility group box), 利用RACE-PCR技术从SMART cDNA文库中克隆到一个Sox基因的cDNA全长, 通过Clustal X和MEGA 4软件对该序列进行比对分析并构建系统进化树; 通过荧光定量PCR技术对该基因在不同组织及发育不同时期性腺中的表达情况进行分析。结果显示, 马氏珠母贝该Sox基因的cDNA全长为1579 bp, 其中开放阅读框(ORF)为1008 bp, 编码336个氨基酸, 5'非编码区为126 bp, 3'非编码区为445 bp。同源性分析表明, 马氏珠母贝Sox基因与太平洋牡蛎Sox11基因的同源性(Identity)最高, 为80%, 故命名为pmSox11; 系统进化树分析也显示pmSox11与太平洋牡蛎Sox11基因的亲缘关系最近。荧光定量PCR分析组织表达特异性显示, pmSox11在马氏珠母贝神经节分布较多的组织如外套膜、鳃、足、消化盲囊等大量表达, 在神经节相对较少的闭壳肌和卵巢中表达量较少; 时序表达图谱显示, pmSox11在3月龄幼贝性腺和1年龄发育早期精巢中表达量最高, 在2年龄成熟精巢和2年龄性转换性腺中表达量降低, 而在2年龄卵巢中表达量最低。研究表明, pmSox11基因可能在马氏珠母贝早期神经系统发育和性别发育的调控方面起到重要作用。     

马氏珠母贝前列腺素E合酶2基因PGES-2的克隆与表达分析

前列腺素E合酶2 (prostaglandin E synthase, PGES-2)是一种核周蛋白,是前列腺素E_2(prostaglandin E_2, PGE_2)合成的末段限速酶之一。研究发现,PGES-2通过与上游诱导酶偶合形成通路调节PGE_2产生方式参与有机体炎症反应、神经系统疾病、促进组织溃疡等各种病理生理事件。为探索马氏珠母贝前列腺素E合酶2 (Pinctada fucata martensii, Pm-PGES-2)的序列特征及其表达情况,本实验通过RACE技术克隆出马氏珠母贝PGES-2基因的cDNA全长序列(Pm-PGES-2),并通过生物信息学软件分析该序列的功能结构;运用实时荧光定量技术检测马氏珠母贝PGES-2基因在各组织中的相对表达量。实验结果显示马氏珠母贝PGES-2基因cDNA序列全长1 419 bp,其中开放阅读框993 bp,编码330个氨基酸,非翻译区5 UTR长170 bp,3 UTR长256 bp。荧光定量PCR检测结果显示该基因在肝胰腺中表达量最高,鳃、边缘膜次之,各组织表达量差异具统计学意义(p0.05)。本研究为进一步探究PGES-2在马氏珠母贝中的生物学功能提供研究基础。     

Plant-Parasitic Nematodes of South Viet Nam

Between 1974 and 1978, 2,842 identifications of plant-parasitic nematodes were made from more than 1,700 soil and plant samples collected in eight provinces of South Viet Nam. Species in nine genera—Helicotylenchus, Criconemoides, Meloidogyne, Pratylenchus, Tylenchorhynchus, Hoplolaimus, Hirschmanniella, Xiphinema, and Rotylenchulus—comprised 96.1% of the identifications; the remaining 3.9% were species of 11 genera. Fourteen genera were associated with rice which was grown on about 2,500,000 ha in 1970. Of these, Ditylenchus, Hirschmanniella, and Meloidogyne were most important. Ditylenchus angustus caused severe damage to about 50,000 ha of flooded rice in the Mekong Delta in 1976. Hirschmanniella spp. were found in all samples examined from flooded rice fields. Meloidogyne spp. were common in rice seedbeds, upland rice, and rice not kept flooded continuously. Meloidogyne and Pratylenchus spp. were found in roots of 22 of the 32 crop plants sampled. Little or no attempt was made in South Viet Nam to control nematodes.     

Study of types of some species of "Filaria" (Nematoda) parasites of small mammals described by Von Linstow and Molin

Parasitic nematodes from the Berlin (ZMB) and Vienna (NMW) Museum collections referred to the genus Filaria Mueller, 1787 by von Linstow or Molin were studied. Three samples were in good condition and the specimens redescribed. Litomosa hepatica (von Linstow, 1897) n. comb., sample ZMB Vermes Entozoa 3368, from the megachiropteran Pteropus neohibernicus, Bismarck Archipelago, resembles L. maki Tibayrenc, Bain & Ramanchandran, 1979, from Pteropus vampyrus, in Malaysia, but the buccal capsule differs. Both species display particular morphological characters which differ from species of Litomosa parasitic in microchiropterans. The remaining material originates from Brazil. The spicule morphology of Litomosoides circularis (von Linstow, 1899) Chandler, 1931, sample ZMB Vermes Entozoa 1059 from Hesperomys spec. (= Holochilus brasiliensis), Porto Alegre, confirms that it belongs to the sigmodontis group; the microfilaria presents characters of the genus Litomosoides, e.g. body attenuated at both extremities and salient cephalic hook. Taxonomic discussions by others confirm that species of Litomosoides belonging to the sigmodontis group and described subsequently are distinct from L. circularis. Litomosoides serpicula (Molin, 1858) Guerrero, Martin, Gardner & Bain, 2002, is redescribed, sample NMW 6323 from the bat Phyllostoma spiculatum (= Sturnira lilium), Ypanema. It is very close to L. brasiliensis Almeida, 1936, type host Moytis sp., but distinguished by a single ring in the buccal capsule, rather than two, supporting previous conclusions that the taxon L. brasiliensis, as generally regarded, may represent a complex of species. Samples NMW 6322 and NMW 6324, from other bats and also identified by Molin (1858) as Filaria serpicula, contain unidentifiable fragments of Litomosoides incertae sedis. Filaria hyalina von Linstow, 1890, sample ZMB Vermes Entozoa Q 3905 from Sorer vulgaris (= Sorex araneus), is incertae sedis because it contains two unidentifiable posterior parts of male, which might be an acuarid, Stammerinema sp. Filaria vesperuginis von Linstow, 1885, sample ZMB Vermes Entozoa Q 3929, from the bat Vesperugo serotinus (= Eptesicus serotinus), contains encysted nematode larvae and is a nomen dubium.     

Multilocus phylogenetic analysis of the genus Aeromonas

A broad multilocus phylogenetic analysis (MLPA) of the representative diversity of a genus offers the opportunity to incorporate concatenated inter-species phylogenies into bacterial systematics. Recent analyses based on single housekeeping genes have provided coherent phylogenies of Aeromonas. However, to date, a multi-gene phylogenetic analysis has never been tackled. In the present study, the intra- and inter-species phylogenetic relationships of 115 strains representing all Aeromonas species described to date were investigated by MLPA. The study included the independent analysis of seven single gene fragments (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD), and the tree resulting from the concatenated 4705 bp sequence. The phylogenies obtained were consistent with each other, and clustering agreed with the Aeromonas taxonomy recognized to date. The highest clustering robustness was found for the concatenated tree (i.e. all Aeromonas species split into 100% bootstrap clusters). Both possible chronometric distortions and poor resolution encountered when using single-gene analysis were buffered in the concatenated MLPA tree. However, reliable phylogenetic species delineation required an MLPA including several “bona fide” strains representing all described species.     

Fate map of the distal portion of Drosophila proboscis as inferred from the expression and mutations of basic patterning genes

The late-third-instar labial disc is comprised of two disc-proper cell layers, one representing mainly the ventral half of the anterior compartment (L-layer) and the other, the dorsal half of the anterior compartment and most, if not all, of the posterior compartment (M-layer). In the L-layer, Distal-less represses homothorax whereas no Distal-less-dependent homothorax repression occurs in the M-layer where Distal-less is coexpressed with homothorax. In wild-type labial discs, clawless, one of the two homeobox genes expressed in distal cells receiving maximum (Decapentaplegic+Wingless) signaling activity in leg and antennal discs, is specifically repressed by proboscipedia. A fate map, inferred from data on basic patterning gene expression in larval and pupal stages and mutant phenotypes, indicates the inner surface of the labial palpus, which includes the pseudotracheal region, to be a derivative of the distal portion of the M-layer expressing wingless, patched, Distal-less and homothorax. The outer surface of the labial palpus with more than 30 taste bristles derives from an L-layer area consisting of dorsal portions of the anterior and posterior compartments, each expressing Distal-less. Our analysis also indicates that, in adults and pupae, the anterior-posterior boundary, dividing roughly equally the outer surface of the distiproboscis, runs along the outer circumference of the inner surface of distiproboscis.     

Microbial transformation of zaluzanin-D

Microbial transformation of zaluzanin-D using different fungi gave 11,13-dihydrozaluzanin-C, zaluzanin-C, 4,16,11,13 - tetrahydro zaluzanin-C, estafiatone, dihydroestafiatol and dihydroestafiatone.     

Molecular phylogeny of Phoma and allied anamorph genera: Towards a reclassification of the Phoma complex

The present generic concept of Phoma is broadly defined, with nine sections being recognised based on morphological characters. Teleomorph states of Phoma have been described in the genera Didymella, Leptosphaeria, Pleospora and Mycosphaerella, indicating that Phoma anamorphs represent a polyphyletic group. In an attempt to delineate generic boundaries, representative strains of the various Phoma sections and allied coelomycetous genera were included for study. Sequence data of the 18S nrDNA (SSU) and the 28S nrDNA (LSU) regions of 18 Phoma strains included were compared with those of representative strains of 39 allied anamorph genera, including Ascochyta, Coniothyrium, Deuterophoma, Microsphaeropsis, Pleurophoma, Pyrenochaeta, and 11 teleomorph genera. The type species of the Phoma sections Phoma, Phyllostictoides, Sclerophomella, Macrospora and Peyronellaea grouped in a subclade in the Pleosporales with the type species of Ascochyta and Microsphaeropsis. The new family Didymellaceae is proposed to accommodate these Phoma sections and related anamorph genera. The present study demonstrated that Phoma radicina, the type species of Phoma sect. Paraphoma and Phoma heteromorphospora, the type species of Phoma sect. Heterospora can be assigned to the Phaeosphaeriaceae and Leptosphaeriaceae respectively.     

Molar microwear and dietary reconstructions of fossil cercopithecoidea from the Plio-Pleistocene deposits of South Africa

The South African Plio-Pleistocene cave deposits have yielded a diverse cercopithecoid fauna. In this study, the possible dietary proclivities of these extinct species are examined using details of molar microwear. Although sample sizes are often small, wear patterns suggest possible temporal changes in the diets of Parapapio jonesi from Makapansgat to Sterkfontein, of Papio robinsoni from Sterkfontein to Swartkrans, and Cercopithecoides williamsi from Makapansgat to Sterkfontein to Swartkrans. However, there does not appear to have been a significant change in the dietary habits of Parapapio broomi over time. The microwear patterns of the two temporally successive congeners, Theropithecus darti and T. oswaldi show no significant differences from one another. The sympatric congeners, Parapapio broomi and Pp. jonesi, have microwear signatures that differ significantly at Makapansgat (Members 3 and 4) but not at Sterkfontein (Member 4). Finally, the microwear analyses suggest that the extinct cercopithecoid species did not necessarily have diets similar to those of their closest living relatives.