三角帆蚌生长性状和内壳色与所产无核珍珠质量的相关性分析

为了研究供片蚌和育珠蚌对无核珍珠质量的影响,以三角帆蚌紫色选育系F₅为材料,在插植无核珍珠前,测量了供片蚌和育珠蚌的壳长、壳高、壳宽、体质量等生长性状,检测了供片蚌内壳色颜色参数L、a、b、dE。经过18个月育珠,测量了育珠蚌的壳长、壳高、壳宽、体质量,计算其特定生长率,检测了育珠蚌内壳色颜色参数L、a、b、dE,测量了育珠蚌所产无核珍珠的颜色、大小、圆度、光泽和产珠量。结果发现,供片蚌生长性状与所产无核珍珠大小、圆度和产珠量相关性不显著(P>0.05)。供片蚌内壳色与无核珍珠颜色相关性极显著(r=-0.400~0.376,P<0.01),供片蚌dE值越大所产紫色珍珠比例越高、dE值越小所产白色珍珠比例越高。育珠蚌特定生长率与所产无核珍珠大小、光泽和产珠量相关性极显著(r=0.237~0.516,P<0.01),相关程度为体重> 壳长> 壳宽> 壳高,育珠蚌特定生长率与所产无核珍珠圆度相关性极显著(r=-0.284~-0.256,P<0.01),相关程度为壳长>体质量>壳宽>壳高。育珠蚌内壳色与所产无核珍珠颜色、大小、圆度、光泽和产珠量相关性不显著(P>0.05)。综合各性状相关性分析,改良供片蚌内壳色可以改良珍珠颜色,改良育珠蚌的生长性状可以改良珍珠大小、光泽、圆度和产珠量。     

三角帆蚌EFCB1基因cDNA序列克隆及表达研究

为了发掘更多三角帆蚌具有EF-hand结构域的功能基因及其蛋白质,本研究运用RACE-PCR技术,克隆得到了三角帆蚌包含EF-hand结构域钙结合蛋白1基因(EF-hand calcium-binding domain-containing protein 1,EFCB1)的cDNA全长并进行了生物信息学分析;通过real-time Q-PCR技术,分析了EFCB1基因在三角帆蚌10个组织,以及内脏团、外套膜插核后不同时间点的时空表达特点。结果表明三角帆蚌EFCB1基因cDNA序列全长981 bp,ORF为531 bp,编码176个氨基酸残基,5'-UTR 239 bp和3'-UTR 211 bp。EFCB1分子式为C₈₇₇H₁₃₄₈N₂₃₈O₂₇₀S₁₀,分子量约19.9 ku,等电点为4.70,不稳定系数为62.65,属亲水蛋白。其序列无信号肽序列,存在1个跨膜区域和2个EF-hand结构域,EF-hand模块分别为DLNDDKLISPEE(98-109)和DTNGDDKLDGEE(129-140)。荧光定量结果显示三角帆蚌EFCB1基因在各组织中均有表达,其中在肠和鳃中表达量最高(P<0.05),外套膜中表达量显著高于内脏团(P<0.05)。EFCB1基因在插核后不同时期的外套膜和内脏团育珠部位组织中表达具有显著差异(P<0.05),在外套膜中的表达量均显著高于内脏团(P<0.05),在插核后第20 天时表达量显著高于各时期(P<0.05)。研究表明,EFCB1在三角帆蚌Ca²⁺的吸收过程中发挥调节作用,在珍珠囊形成过程中以及珍珠形成初期具有重要功能。     

三角帆蚌HcCUBDC基因cDNA的全长克隆与表达分析

利用cDNA末端快速扩增（Rapid Amplification of cDNA Ends,RACE）方法,获得了三角帆蚌HcCUBDC基因的全长cDNA序列,共5158bp,开放阅读框（ORF）1920bp,编码639个氨基酸,5′端非编码区576bp,3′端非编码区2662bp,GeneBank登陆号为KP067952。生物信息学分析表明,三角帆蚌HcCUBDC蛋白含有一个由19个氨基酸组成的信号肽和4个CUB结构域,但未能比对上已知的蛋白。经荧光定量PCR检测,HcCUBDC基因在紫色和白色蚌前端缘膜、后端缘膜、中央膜、鳃、斧足、肝胰腺、肠和肾8个组织中均有表达,并且都是肝胰腺表达量最低。在紫色蚌中,后端缘膜表达量极显著高于前端缘膜;在白色蚌中,前端和后端缘膜之间表达差异不显著。HcCUBDC基因在紫色蚌后端缘膜中的表达量极显著高于白色蚌,在紫色和白色蚌前端缘膜中的表达量差异不显著。外套膜原位杂交结果显示,HcCUBDC基因主要在缘膜外上皮细胞中表达。研究表明,HcCUBDC基因在三角帆蚌内壳色形成中发挥作用,可为进一步深入研究该基因在珍珠颜色形成过程中的功能及其调控机理提供基础资料。     

三角帆蚌种质资源研究进展

三角帆蚌是我国优良的淡水育珠蚌,具有重要的经济价值。本文从我国三角帆蚌种质资源调查、搜集与保护出发,介绍了三角帆蚌RAPD、SSR及线粒体基因片段等不同分子标记开发状况,重点讨论了这些分子标记分析三角帆蚌遗传多样性的情况。比较了我国五大淡水湖泊三角帆蚌种质及3个优秀种质9个F1后代的生长性能,比较了三角帆蚌雌雄生长差异。简述了三角帆蚌珍珠质形成相关基因、免疫相关基因、贝壳及珍珠颜色相关基因的研究现状。从三角帆蚌与其他蚌类种间杂交、三角帆蚌种内杂交,介绍了培育康乐蚌新品种的过程及三角帆蚌家系选育的进展情况。对今后进一步开展三角帆蚌种质资源研究提出了意见和建议,为三角帆蚌遗传改良提供了基础资料。     

Freshwater nucleated pearl quality is influenced by host mussel growth traits in Hyriopsis cumingii

Hyriopsis cumingii is one of the most important freshwater pearl mussels in China. Recently, this species can produce freshwater nucleated pearls of high quality. Here, we investigated whether nucleated pearl quality is influenced by the growth traits of the host mussel or other factors like cultivation period. We implanted host mussels with a spherical nucleus consisting of a small piece of mantle tissue from donor mussels. After 24 and 36 months of culture, host mussel growth traits including body weight and shell length, height, width and weight were recorded. These factors were then correlated with the quality traits of the pearls they produced, such as nacre thickness, size, weight, lustre and colour. Results indicated pearls obtained at 36 months after seeding were significantly larger in terms of nacre thickness, size and weight compared to those harvested at 24 months. In particular, nacre thickness (r = 0.33–0.48, P = 0.00), pearl size (r = 0.39–0.43, P = 0.00) and pearl weight (r = 0.35–0.47, P = 0.00) were showed to be significantly correlated with host mussels shell length, body weight and shell weight at 24 or 36 months. Larger and heavier host mussels tended to produce bigger pearls. In contrast, host mussels did not affect pearl colour. Cumulatively, our results suggest that longer culturing times and a larger host mussel may help produce better quality nucleated pearl. This information can help guide selective breeding programs designed to improve pearl quality produced by H. cumingii.     

Comparison of growth and pearl production in males and females of the freshwater mussel, Hyriopsis cumingii, in China

Growth and pearl production were compared for males and females in the freshwater mussel, Hyriopis cumingii, from a full-sib family. The results indicated that there was no significant difference (p > 0.05) on individual weight between male and female mussels of 1- or 2-year-old, while significant difference (p < 0.01) lay among 3- or 4-year-old mussels with male greater than female. The average shell width of the male mussels was less than that of the female individual (p < 0.05). 1- and 2-year-old males and females did not differ significantly with respect to the total weights, grain weights, or grain sizes of the pearls they produced, but these three parameters were all significantly greater in 3- and 4-year-old males (p < 0.05). The round pearl percent were similar between male and females at ages 1, 2, 3 and 4 years. Male and female mussels were separately and mixed cultured in enclosures, respectively. The rates of growth in shell width and body weight of females separated from males were 3.42 and 4.16 %, respectively, higher than in females mixed with males (p < 0.05). The total pearl weight per mussel, the average weight per pearl, and the average pearl size of females separated from males were 6.61, 7.10, and 3.59 %, respectively, greater than in females mixed with males (p < 0.05). There was no significant difference in the growth rate or pearl yield of males cultured with or without females (p > 0.05). Under traditional culture methods, male mussels have a better pearl performance, and artificial separation of females from males can improve the growth and pearl production of female mussels.     

缺氧和有毒微囊藻胁迫下三角帆蚌鳃和主要消化器官以及晶杆体的扫描电镜观察

为了阐明缺氧和有毒铜绿做囊藻对三角帆蚌鳃和主要消化器官的毒理效应和组织病理,实验利用扫描电镜研究了暴露于缺氧和有毒铜绿做囊藻下三角帆蚌的鳃、胃、肠以及晶杆体等器官组织病理学变化。结果显示,从第7天开始缺氧和有毒藻类复合胁迫组三角帆蚌的鳃丝及柱状细胞出现大量坏死和脱落,胃肠腔面纤毛脱落、上史细胞破裂坏死,晶杆体在第5天已经完全消失;第7天单独缺氧组和单独有毒藻类组的鳃和消化道出现少量的病变,单独缺氧组晶杆体在第7天完全消失,单独有毒藻类组晶杆体则一直存在。胁迫消失后,3个实验暴露组的晶杆体都未恢复正常对照组水平,因此缺氧和有毒铜绿做囊藻对三角帆蚌鳃和消化系统产生不可恢复性损伤,双重胁迫组影响最为严重,缺氧胁迫组相对有毒铜绿做囊藻胁迫组影响更为显著。本研究为三角帆蚌在缺氧和有毒藻类胁迫下的生理适应机制提供了组织病理学上的参考,并为其作为富营养化水体治理工具种的可行性提供了理论依据。     

5种壳色长牡蛎不同组织中金属元素的分析与评价

为评估不同壳色长牡蛎金属元素价值,实采用电感耦合等离子体原子发射光谱仪(ICP-OES)和近红外(NIR)分析模型对5种壳色(壳黑、壳紫、壳橙、壳金和壳白)长牡蛎4个组织(外套膜、鳃、闭壳肌和性腺—内脏团)中的Mg、Fe、Zn、Cu、Mn和Se元素含量进行检测分析,并比较与对照群体金属元素含量的差异以及5种壳色长牡蛎各组织间含量的差异。结果发现,在外套膜中,壳紫、壳橙和壳黑长牡蛎中Zn含量显著高于对照群体,壳紫长牡蛎Cu含量显著高于壳金和对照群体,壳金长牡蛎Mn含量显著高于壳紫和壳黑长牡蛎,其他元素在各群体中含量无显著差异;在鳃中,壳金长牡蛎Mn含量显著高于壳黑和对照群体,其他组份在6个群体无显著性差异;在闭壳肌中,壳橙和壳紫长牡蛎Zn的含量显著高于对照群体,其他组份在各群体中差异不显著;在性腺—内脏团中,壳橙长牡蛎Zn和Mn含量均显著高于对照群体,且Cu的含量显著高于壳金长牡蛎,其他元素在6个群体中含量无显著性差异。4个组织中各金属元素含量差异显著,其中闭壳肌内各金属元素含量显著低于鳃、外套膜和性腺—内脏团。研究表明,不同壳色长牡蛎之间以及不同壳色与对照群体之间在部分金属元素含量上已表现出分化,这为长牡蛎壳色新品系选育提供了重要的基础资料。     

插片后珍珠蚌生长性状及其无核珍珠成珠率和大小

为了筛选优质的珍珠蚌插片组合,探究不同种类珍珠蚌相互插片后生长性状间的差异和相关性,以及各组合无核珍珠成珠率和大小的差异,利用三角帆蚌(S)、池蝶蚌(C)和褶纹冠蚌(Z)分别作为供片蚌和育珠蚌,获得9个育珠蚌组合,测量各育珠组合的体质量、壳长、壳高和壳宽,比较9个育珠蚌组合及3个未插片的三角帆蚌、池蝶蚌和褶纹冠蚌对照组在1年后生长性状的差异,对各生长性状相关性进行分析。测量每个育珠蚌所产无核珍珠的数量和大小,计算成珠率和圆度,分析不同组合之间无核珍珠的差异。结果显示,无核育珠手术影响珍珠蚌的生长性能,以三角帆蚌和池蝶蚌为育珠蚌的组合生长性状均优于对照组,S-S育珠组合在以三角帆蚌为育珠蚌组合内生长最优,SC和C-C育珠组合在以池蝶蚌为育珠蚌组合中生长最优,而以褶纹冠蚌为育珠蚌的组合插片后生长性状指标均小于对照组。其中除了池蝶蚌同种插片组合(C-C育珠组合)外,其余各育珠组合壳长与体质量间相关系数均高于其他生长性状间相关系数,C-C育珠组合壳宽与体质量相关系数最大;三角帆蚌和池蝶蚌之间的插片组合(S-S、C-S、S-C和C-C育珠组合)及褶纹冠蚌同种插片组合(Z-Z育珠组合)成珠率较高(...     

Evaluation of laser labeling on growth and survival of the freshwater pearl mussel, Hyriopsis cumingii

Laser labeling techniques have been developed for breeding programs in aquatic organisms through this study. This study assessed their use in Hyriopsis cumingii, an important Chinese freshwater pearl mussel. In preliminary studies, H. cumingii juveniles between 55 and 95?days old were labeled by a laser engraving machine. The mortality rate decreased in an age-dependent manner, no differences in growth between treated and unlabeled juveniles at 85 and 95?days of age. Hundred-day-old juveniles were labeled with four different sized labels. The mortality rate and growth inhibition were significantly higher with the largest label size (2.55?cm) compared to the smaller size and control. Four different types of laser intensity were applied to 100-day-old juveniles, but mortality rates and growth inhibition after high current treatments were significantly higher than the lower currents and control. The wing part, center, lower edge, left edge and right edge were labeled in different 100-day-old juveniles. Labeling of the left edge caused higher mortality and growth inhibition than the other sites. No significant differences were found after either side of the shell was labeled compared to the control. All labels remained clear without any deformation after several months. In summary, the optimal strategy for the laser labeling of H. cumingii would be in juveniles >85?days. The label size, laser intensity and positions could affect their growth and survival. There was no difference labeling either side of the shell. This study showed laser labeling is a relatively simple and practical marking method for mussels such as H. cumingii.     

Effects of integrated combination and quicklime supplementation on growth and pearl yield of freshwater pearl mussel, Hyriopsis cumingii (Lea, 1852)

The effects of integrated combination and quicklime supplementation on growth and pearl yield of a freshwater mussel, Hyriopsis cumingii (Lea, 1852), were examined through a 137-day growout in land-based enclosures. The integrated combinations examined were either mussel, bighead carp and gibel carp or mussel and bighead carp. Each combination was treated either with or without quicklime supplementation. One half of the mussels in each enclosure were grafted with pieces of the mantle epithelium while the other half were not. During the experiment, gibel carp were fed formulated feed while the mussel and bighead carp were fed natural live food. Quicklime was regularly provided in the enclosures as calcium replenishment. The species composition in the integrated system significantly affected growth in shell size and wet weight of the mussels regardless of the graft and pearl yield, while no significant effects of quicklime supplementation were detected. Growth rates in shell size and wet weight of both grafted and non-grafted mussels and pearl yield were slightly higher in the enclosures with mussel, bighead carp and gibel carp than those with mussel and bighead carp, although these differences were not statistically significant. The non-grafted mussel exhibited faster growth in shell size and wet weight than the grafted mussel within the same treatment. Results of the present study indicate that species combination in an integrated system can affect growth and pearl yield of H. cumingii . The species combination of mussel, bighead carp and gibel carp is recommended for commercial H. cumingii farming.     

Freshwater Pearl Culture Technology Development in India

ABSTRACT

Development of science and technology of cultured pearl production in a freshwater environment in India is described. Distribution of Indian pearl mussels, pond mussel Lamellidens marginalis, paddy field mussel L. corrianus, and riverine mussel Parreysia corrugata in relation to environmental variables such as type of soil, nature of sediment substratum, presence or absence of macrophytes (Eichhornia sp., Nechamandra sp., andNymphaea sp.) is described. Food and feeding of the mussels, together with density aspects in culture conditions are discussed. Mussel breeding including specificity in fish hosts, and mussel larval parasitic relationships are discussed. Basic steps involved in indigenous freshwater pearl culture technology are summarized. Pearl culture grafting procedures such as mantle cavity, mantle tissue and gonadal implantation along with different pearl products are also described.     

企鹅珍珠贝组织蛋白酶D的cDNA克隆、序列特征分析和应激表达研究

笔者初步研究了企鹅珍珠贝（Pteria penguin）组织蛋白酶D（cathepsin D,CTSD）的基因克隆和功能,通过同源克隆方法和cDNA末端快速扩增技术（RACE）获得了企鹅珍珠贝组织蛋白酶D基因（命名为pgCTSD）。该基因cDNA全长1 767 bp,其中5＇UTR为38 bp,3＇UTR为553 bp,ORF为1 176 bp,编码392个氨基酸,包括信号肽（Met1-Ala18）、前体域（Leu19-Lys47）和成熟域（Tyr48-Ser392）三部分,分子量为42.3 kDa,等电点为8.04。pgCTSD氨基酸序列与大珠母贝（Pinctada maxima）pmCTSD的相似性最高（79%）,与其他物种的相似性为59%～75%。荧光定量分析表明,空白对照组中pgCTSD mRNA在闭壳肌、性腺、肝胰脏、外套膜和鳃组织中都有表达,且在闭壳肌中表达量最少,肝胰脏中最高。与试验对照组相比,脂多糖（LPS）刺激6 h后性腺和肝胰脏显著下降,闭壳肌的表达量虽不大但增加显著,外套膜和鳃组织变化不显著;哈维弧菌（Vibrio harveyi）刺激6 h后肝胰腺和外套膜显著下降,闭壳肌和鳃显著上升,性腺无显著变化。肝胰腺中pgCTSD对LPS和弧菌刺激的应答反应表明pgCTSD可能参与了免疫反应。     

Variation of Color and Ray Pattern in Juvenile Shells in Hatchery‐produced Freshwater Triangle Pearl Mussels,Hyriopsis cumingii,in China

The variation and inheritance of juvenile shell color and pigmentation pattern in the freshwater mussel, Hyriopsis cumingii, were reported in 1‐yr‐old progeny of eight families. There were three distinctive phenotypes of shell color and pigmentation patterns observed, including a greenish‐brown shell with radial rays, yellowish‐brown shell with radial rays, and yellowish‐brown shell without radial rays. There were no greenish‐brown individuals without radial rays. The shell color phenotypes showed variation with the growth in juvenile Stage I (1–3 cm in shell length), and the percentage of individuals with radial rays increased once they reached a shell length of 11 mm and then stabilized after reaching 20 mm in shell length. Shell color differentiation became more apparent at a shell length of 26 mm. Results of chi‐square tests of the segregation ratio of shell color or ray phenotypes obtained from eight families at juvenile Stage II (6–9 cm in shell length) suggested that greenish‐brown is controlled by a dominant allele (G) and yellowish‐brown‐shell phenotype is by a recessive allele (y); the ray pattern phenotype is controlled by a recessive (r) and a dominant allele (R) at a single locus. Shell color phenotypes may be a useful genetic marker for future selective breeding of triangle pearl mussels.     

背角无齿蚌抗菌肽theromacin基因cDNA全序列克隆及表达分析

采用RACE方法,克隆了背角无齿蚌抗菌肽theromacin基因的cDNA全序列。结果显示,该cDNA序列全长为870 bp,5'端非翻译区104 bp,3'端非翻译区460 bp,开放阅读框长为306 bp,共编码101个氨基酸,相对分子量为11 166.9 u。同源性分析显示,该基因编码的蛋白与三角帆蚌theromacin基因氨基酸序列的相似度最高,为73%;与贻贝中发现的抗菌肽defensins、mytilins、myticins和mytimycins等4种类型相似度较低,属于Macin家族(登录号:KJ598604)。实时荧光定量PCR分析结果显示,该基因在血液、肝脏、外套膜、鳃、斧足和肠等组织中均有表达,在外套膜中的表达量最高,在其他组织中的表达量较低;经嗜水气单胞菌诱导后,各个组织的表达量出现明显上调的趋势且与对照组显著差异,推测theromacin基因可能在背角无齿蚌的免疫反应中起重要作用。     

An evaluation of different methods for transportation of the freshwater mussel <Emphasis Type="Italic">Lamellidens corrianus</Emphasis> (Lea 1834)

The freshwater mussel Lamellidens corrianus (Lea 1834) is widely distributed in ponds and large bodies of perennial waters in the Indian subcontinent. It is one of the important species for producing freshwater pearls in India. As the freshwater pearl aquaculture activity may expand on wide scale in future, it may require collecting L. corrianus from the distant places and transport to pearl producing centres. Hence, in order to develop suitable method for transportation of L. corrianus to obtain high survival with minimum physiological stress, the present study was conducted. L. corrianus were transported by two different methods i.e. in air (in wet gunny bags) and in water (in plastic polyethylene bags containing water) to find out the best method for transportation. Glucose was chosen as the stress factor as the heightened circulating glucose levels may reflect the bivalve’s ability to respond to weather extremes, handling stress or any recent environmental stress perceived to require extra energy. Experiments were conducted both under field as well as laboratory conditions, during the start of winter season, where regional temperature falls to 4–5°C below the average. Under the field experiment, L. corrianus (total length = 8.26 ± 0.34 cm; wet weight = 52.62 ± 3.18 g) were transported by road from Khopoli (Dist. Raigad, Maharashtra State) to Ratnagiri (Dist. Ratnagiri, Maharashtra State) for 12 h in air and water (reservoir water temperature = 26.4°C). Under laboratory conditions, simulating air transportation was done in wet gunny bags (42.5 cm × 62.5 cm), each bag containing 50 mussels (total length = 8.34 ± 0.46 cm; wet weight = 53.2 ± 3.24 g). Simulating water transportation was carried out using plastic polyethylene bags (45 cm × 60 cm), with 50 mussels in five litres of water (water temperature = 26.5°C). At periodical intervals, glucose concentrations from mantle, gill and posterior adductor muscle were estimated. Under both field and laboratory conditions, glucose levels were significantly higher in mantle, gill and posterior adductor muscle tissue of L. corrianus transported in air when compared to that of water-transported mussels. Results showed that air transportation was comparatively more stressful than water transportation. However, cent per cent survival was obtained in the both methods just after transportation and after 7 days of transportation. Considering the absence of mortality in both methods, it can be concluded that the choice of the mussel transportation method is more economical dependent than biological dependent.     

三角帆蚌hcSRCR1基因的克隆及在不同壳色选育系中的表达模式

清道夫受体(SR)是一类对化学修饰的脂蛋白具有很强结合活性的糖蛋白家族。本研究通过RACE方法克隆得到三角帆蚌hcSRCR1基因cDNA序列,该序列全长1 000 bp,其中开放阅读框819 bp,编码272个氨基酸,预测分子量为28.16 ku,理论等电点为5.55;预测含有2个SRCR结构域和6个保守的半胱氨酸残基。qRT-PCR和Western blot结果显示,hcSRCR1 mRNA和蛋白表达模式基本相同,均在三角帆蚌外套膜中表达量最高,在其他组织中的表达量普遍较低,且在紫色选育系外套膜组织中的表达量显著高于白色选育系。外套膜原位杂交结果显示,hcSRCR1基因主要在外套膜外褶的内、外上皮细胞层以及腹膜处的上皮细胞层中表达。研究表明,三角帆蚌hcSRCR1基因与贝壳珍珠质颜色形成具有一定相关性,可为进一步研究该基因在珍珠颜色形成过程中的调控机理提供基础资料。     

不同施肥方法对鱼蚌综合养殖池塘浮游植物群落结构的影响

通过155 d围隔实验检验了不同施肥条件下鱼蚌综合养殖水体中的浮游植物群落结构。实验设3个处理:施鸭粪、施化肥、兼施鸭粪和化肥。放养种类为三角帆蚌、草鱼、鲫、鲢和鳙,放养量分别为每围隔20、15、5、5和5个。结果发现,围隔内浮游植物生物量平均值为(2.1~6.0)×108个/L。不同施肥方法对浮游植物种类组成和优势种、叶绿素a(Chl.a)、生物量以及蓝藻在浮游植物生物量中的比例无显著影响。浮游植物群落变化表现出较明显的季节性特点,影响围隔浮游植物群落的理化因子为TN、NH3-N和DO。研究表明,采用不同施肥方法的围隔内浮游植物群落结构未表现出显著差异,难以从浮游植物角度解释兼施鸭粪和化肥的围隔珍珠产量高于施鸭粪或施化肥的围隔的事实,也难以确定珍珠产量与浮游植物群落结构之间存在必然的联系。     

转录组测序研究三角帆蚌珍珠颜色相关基因

为筛选影响三角帆蚌珍珠颜色的候选基因,采用转录组测序平台Illumina Hi Seq TM2500对蚌壳珍珠层颜色为紫色和白色的三角帆蚌中央膜组织进行高通量测序。所得序列经质控、组装后比对到NR、Swiss-Prot、COG、KOG、KEGG、GO和Pfam数据库中注释,并进行差异基因聚类分析。结果显示,获得干净数据共72 800 581 bp,拼接获得了89 529个Unigene,差异表达基因2644个,其中上调表达基因1323个,下调表达基因1321个。根据GO功能分类可分为分子功能、细胞组分、生物过程等3大类50分支;根据KEGG代谢通路分析可以分为187类。对差异表达基因分析发现,部分基因与色素合成(眼黄素、黑色素、喋啶色素)及金属离子转运相关,还有6个细胞色素P450,3个细胞色素b561和1个细胞色素b560。研究表明,这些基因可能在珍珠颜色形成中发挥作用。     

体外培养条件下三角帆蚌胚胎早期发育观察及胚胎发育生物学零度和有效积温分析

为了探寻三角帆蚌胚胎体外培养方法,本实验以解剖获得的雌雄配子为材料,使用与三角帆蚌体液等渗的平衡盐溶液(BSS)进行人工体外受精和体外培养,观察各阶段胚胎的形态特征,并分析计算不同胚胎发育阶段的生物学零度和有效积温。结果显示,在温度为(25±1)℃的BSS培养条件下,受精卵发育至第二极体排放、二细胞期、四细胞期、八细胞期、十六细胞期和桑葚期的时间分别为1.8、2.8、5.6、10.5、13.9和17.3 h,胚胎最长发育至桑葚期后停止发育。另外,池塘平均水温分别为26.5、28.1和29.2℃时,三角帆蚌胚胎发育至成熟钩介幼虫分别需要10、9和8 d,计算得出三角帆蚌胚胎发育的生物学零度为14.81℃,根据生物学零度计算胚胎发育至卵裂期、囊胚期、原肠期、膜内钩介幼虫期和成熟钩介幼虫的有效积温分别为12.95、25.99、42.27、69.21和118.14℃×d。研究结果初步尝试对三角帆蚌受精卵进行体外培养,并根据胚胎发育水温和时间获得三角帆蚌早期胚胎发育的生物学零度和有效积温,可为突破三角帆蚌全人工繁育技术奠定基础,对淡水贝类现代生物育种技术研发和种质资源保护具有重要意义。