淡水有核珍珠养殖技术（二）——淡水有核珍珠插植方法

在人们尚未揭示珍珠形成的奥秘前．流行珍珠是“露滴成珠”、“日月的精华”和“鲛人的眼泪”等的说法。后来有人切开珍珠发现核心部分是砂粒等异物．如是便有“异物刺激成珠”的推测。13世纪左右．我国劳动人民发现贝类的外套膜分泌珍珠．并由此而培育了举世闻名的佛像珍珠。1806年．日本“珍珠之父”御木本幸吉氏．把外套膜成珠的机制和“异物刺激成珠”的推测相结合，在贝类软组织上先插植1颗异物(珍珠核)，再往核上移植1块细胞小片，终于从15万只试验贝中．收获了1颗游离的有核珍珠．从而奠定了今日人工插核育珠的技术基础。     

微胶囊饲料对马氏珠母贝育珠性能、生长基因和矿化基因表达的影响

本研究分析了自主研发的微胶囊饲料替代微藻对马氏珠母贝(Pinctada fucata martensii)育珠性能、生长相关基因和矿化相关基因表达的影响。共设置了3个实验组,其中,EG1组投喂亚心形扁藻(Platymonas subcordiformis),EG2组投喂微胶囊饲料+亚心形扁藻,EG3组投喂微胶囊饲料。养殖170 d后,比较马氏珠母贝的育珠性能、闭壳肌生长相关基因EGFR、FGF18、GHITM和TβRI以及外套膜的中央膜和边缘膜矿化相关基因pearlin、DPT、pif177和N19的表达。结果表明:(1)各组间马氏珠母贝育珠贝的存活率、留核率不存在显著性差异(P0.05),所采收珍珠的珍珠层厚度及平均质量也不存在显著性差异(P0.05);(2)各实验组间马氏珠母贝闭壳肌中EGFR、FGF18、GHITM和TβR I的相对表达量不存在显著性差异(P0.05);(3)各实验组间马氏珠母贝中央膜中DPT和N19的相对表达量差异不显著(P0.05),EG2和EG3组pearlin的表达量显著性高于EG1组(P0.05),pif177在EG3组中的表达量显著性高于EG1和EG2组(P0.05);(4)各实验组间马氏珠母贝边缘膜中pearlin、DPT和N19的相对表达量差异不显著(P0.05),EG2和EG3组pif177的表达量显著高于EG1组(P0.05)。研究结果说明,该微胶囊饲料可以替代部分微藻进行育珠生产,为进一步研发马氏珠母贝的人工配合饲料及开展工厂化育珠积累了参考资料。     

三角帆蚌产珠性能与生长性状和插片部位的关系

2003年5月，获得鄱阳湖群体三角帆蚌（Hyriopsis cumingii）同一母蚌的后代，同年10月插种无核珍珠，然后在同一口池塘培育3年，于2006年11月研究其产珠性能与生长性状和插片部位的关系。结果表明，珍珠单颗质量、粒径、直径差百分比、产珠量和圆珠率5个指标的极差以及变异系数非常大，说明同一亲蚌子代个体间在育珠性能方面出现较大分化，可通过选育进行改良。各指标与单蚌产珠量的相关性由高到低依次为，壳质量、体质量、壳高、壳宽、壳长；各指标与珍珠粒径的相关性由高到低依次为，壳质量=体质量、壳宽、壳高；各指标与直径差百分比的相关性由高到低依次为，壳宽、壳质量=体质量；各指标与圆珠率的相关性由高到低依次为，壳宽、壳质量、体质量。结合生产可操作性，体质量和壳宽应作为两个最主要选育性状；位于外套膜外侧的珍珠质量显著高于内侧（P〈0．05），但未发现左壳与右壳间珍珠质量具有显著性差异（P≥，0．05）。[中国水产科学，2008，15（3）；493-499]     

三角帆蚌与池蝶蚌育珠性能比较

三角帆蚌,隶属于瓣鳃纲,真瓣鳃目,蚌科,帆蚌属,是中国特有的优质淡水育珠蚌。三角帆所产的珍珠产量佳,珠质细腻、光滑、色泽鲜艳、形状较圆,但珍珠生长比较缓慢。池蝶蚌与三角帆蚌同属帆蚌属(日本称为池蝶蚌属),是中国大陆近年来从日本、台湾引进的优良珠贝类。与三角帆蚌相比,池蝶蚌具有壳、珍珠层、外套膜较厚,珍珠质分泌能力强等优点。     

影响养殖珍珠质量的主要因子

珍珠被誉为“宝石皇后”,养殖珍珠质量的提升是珍珠产业关注的焦点,也是珍珠研究领域的重要课题.本研究着重介绍了评价养殖珍珠质量的6个方面内容,包括颜色、大小、形状、光泽、光洁度、有核珍珠珠层厚度等.详细陈述了各主要育珠贝种类及其产珠特点,它们所培育的淡水无核珍珠和海水有核珍珠的质量情况,论述了不同规格、不同壳色育珠贝对所产珍珠质量的影响.重点介绍了作为制作小片供体的供片贝不同种类,以及供片贝不同年龄、不同壳色对育珠贝所产珍珠质量的影响.同时,阐述了插片手术过程中,化学药物因素、小片分离方式、插片个数等插片手术工艺对珍珠质量的影响.此外,还从水质、水体微量元素、养殖深度、养殖方式和养殖周期等几个方面系统探讨了外部条件对养殖珍珠质量的影响.     

淡水无核珍珠手术新工艺及操作规程

自从１９１３年奥富德斯和濑辰平、西川藤吉等研究成功采用贝类外套膜上皮组织、移植到育珠贝体内以培育珍珠的方法，至今一直是人工培育珍珠的基本手术工艺。近１００年来，通过许多科学家的不懈努力，使今天的珍珠有了大规模生产的可能，而新的工艺、新的技术不断出现，从而推进全世界育珠业继续向前发展。 中国是世界珍珠生产大国，近几年大陆淡水育珠业又掀起了生产执潮。由于生产的快速发展，影响珍珠质量的诸多问题也随之产生，其中手术操作技术人员紧缺，技术水平低下及管理不善等。目前，国内手术操作普遍存在未经正规培训，操作技…     

企鹅珍珠贝附壳珠规模化培育技术研究

为探索企鹅珍珠贝附壳珠的规模化培育技术,采用"钻孔系线植核法"对企鹅珍珠贝母贝进行植核.植核贝数量为1437834个,育珠期8-12个月,共收获育珠贝1095683个,植核手术育珠期成活率为70.1%-88.3%,平均成活率为84.1%,留核率为100%.共收获附壳珍珠2191366颗,其中商品珠1821025颗,商品...     

河蚌育珠技术讲座

珍球是指由珍珠贝等一些贝类外套膜细胞所分泌的壳角蛋白和碳酸钙结晶围绕一个共同的核心交替沉积而形成的圆形或其它形状的物体。目前我国淡水珍珠的产量据世界首位。产珠地区在逐步扩大,养殖技术在不断改进,销售市场有所发展。但在珍珠生产技术和销售过程中确也存在不少问题,因此     

声音

“育珠贝养殖不走出内湾只有死路一条．如果不是加工厂还有钱赚。从资金上给养殖户一些扶持,内湾育珠这个产业早就岌岌可危了。” ——湛江荣辉珍珠有限公司董事长尹国荣表示．他连续6年无偿提供珠核给养殖户,所用资金达数百万元。内湾育珠死亡率太高了,拿10吨珠核给养殖户插,最终能收获的珍珠连两吨都没有。     

淡水无核珍珠手术新工艺和操作规程

自从 1913年奥富德斯和濑辰平、西川藤吉等研究成功采用贝类外套膜上皮组织，移植到育珠贝体内以培育珍珠的方法，至今一直是人工培育珍珠的基本手术工艺。近 100年来，通过许多科学家的不懈努力，使今天的珍珠有了大规模生产的可能，并且新的工艺、新的技术不断出现，从而推进全世界育珠业继续向前发展。　　经多年努力，我们将撕膜法工艺和系统化消毒技术科学结合，形成了一套新的手术操作工艺、流程，以及手术操作质量管理措施，通过对手术操作工的培训和养殖生产实践表明，该工艺及其管理措施，不仅使“大、光、圆、艳”的优质珠比率…     

Expression profiles of nine biomineralization genes and their relationship with pearl nacre thickness in the pearl oyster,Pinctada fucata martensii Dunker

Pinctada fucata martensii is an ideal animal for study of biomineralization. Although dozens of genes have been identified, the molecular mechanism of biomineralization remains still unclear. The purpose here was to discover the expression profiles of nine biomineralization genes in related tissues: mantle edge (ME), mantle centre (MC) and pearl sac (PS), and to explore the relationships between expression level and nacre thickness. The expression levels of seven genes (ACCBP, aspein, CaM, EFCBP, KRMP, nacrein and prismalin‐14) showed no significant difference between ME and MC, but were significantly higher than those in PS. The expression level of N19 was the same in ME and PS, significantly higher than that in MC. However, msi60 in ME, MC and PS showed no significant differences in expression level. In addition, the correlations of expression levels between ME, MC and PS were highly significant, indicating that there are some similar function and mechanism among the three tissues. Also the nine genes had significant correlation of expression levels between one another. Furthermore, no other genes from oysters which produced pearls of different grades showed significant difference in the expression level, except CaM in MC, which had significantly higher expression level in the medium pearl grade than in the plain nucleus grade. Moreover, the expression level of msi60 in MC of host oyster was significantly correlated with produced pearl layer thickness, suggesting that msi60 might play an important role in pearl formation and host oyster may also participate in the development of pearl.     

合浦珠母贝全同胞家系贝壳珍珠质颜色分析

合浦珠母贝（Pinctada fucata）小片贝的珍珠质颜色直接影响育珠贝所产珍珠的颜色。每个合浦珠母贝的珍珠质颜色都有自己的颜色特征值,即三刺激值和颜色参数。试验选择16个合浦珠母贝全同胞家系,用CSE成像分析色度系统测量并比较各家系左壳内侧外缘珍珠质颜色参数Lab和三刺激值,以筛选适合做插核小片贝的家系。珍珠质的颜色参数明度（L）为98.586～105.234,其中F16家系的明度最大,F3家系的明度最小;a（红绿色品）为-0.967～-6.577,b（黄蓝色品）为5.915～11.237,F14家系色品最偏向于绿色（-a）,F16家系色品最偏向于黄色（b）。F3和F14家系贝壳外缘珍珠质的色差（ΔEab）最大（7.885）,肉眼能明显觉察。16个家系的合浦珠母贝都属于白色系列,其中离白光中心最近（a2＋b2最小）且个体颜色集中的F1家系最适合用作小片贝来选育培养。     

Donor effect on cultured pearl nacre development and shell matrix gene expression in Pinctada margaritifera reared in different field sites

Understanding the respective roles played by donor and recipient pearl oysters in pearl quality determination in relation to the environment is a challenge for the pearl industry. In most Pinctada species, pearl size is mainly related to recipient oyster growth performance but also relies to some extent on the biomineralisation activity of the pearl sac, a tissue that originates from the donor oyster mantle. We examined donor effect on pearl size in response to culture in the lagoons on Arutua and Apataki atolls. Overall, nacre weight and thickness were greater in Arutua than in Apataki, but sensitivity to the environment differed between donors. Some donors were associated with significantly heavier and thicker nacre in Arutua (I group), while others had similar results at the two sites (NI group). On average, up to 20% of the pearl size could be attributed to the donor but, in group I, donor effect was responsible for up to 36% of nacre weight determination. Additionally, a real‐time PCR expression study of eight matrix protein genes related to biomineralisation in the pearl sac showed that MSI60, pearlin and pif177 were significantly and positively correlated with nacre weight and thickness, with the latter two genes explaining the larger pearl size observed in Arutua. Donor oysters in P. margaritifera therefore play a key role in pearl size improvement, related to the role of the shell matrix protein genes. Understanding such contributions could help in the design of genetic selection plans for specific and adapted donor oyster lines.     

Age‐dependence of cultured pearl grade and colour in the black‐lipped pearl oyster Pinctada margaritifera

Pinctada margaritifera is an economically important marine bivalve species for cultured pearl production in French Polynesian aquaculture. In order to evaluate the influence of donor oyster age on pearl quality traits, experiments were conducted over 6 years using both grafts and surgreffe operations. At harvest, six pearl quality traits were recorded and compared: surface defects, lustre, grade, darkness level and visual colour. Analysing the quality traits of pearls harvested in the initial graft process and those of pearls obtained from surgreffe experiments allowed a comparison of the influence of pearl sac cells originating from the initial mantle graft, which aged together with their recipient oysters. The results demonstrated a significant decrease between these successive grafts in lustre, grade (A‐B‐C), darkness level, and green colour – traits that are of major importance in the pearl market. The duplicated graft experiment allowed the comparison of donor oyster families at 2 and 5 years old, where a mantle graft was inserted into recipient oysters aged 2.5 years. The results showed the same tendencies to a lesser extent, with (i) an improved pearl grade, predominantly through a most important rate of 0 surface defect category, and (ii) a green/grey ratio in favour of the younger donor. A comparison between the graft‐surgreffe and the duplicated graft experiments also highlighted: (i) the indirect role played by the younger recipient oysters, which must be optimized for optimal pearl quality realization, and (ii) the complex interplay between the donor and recipient oysters.     

长牡蛎MITF基因表达及与壳色的关联

为了解长牡蛎MITF基因的表达及其与壳色的关联,验证了长牡蛎中的4个MITF基因,对长牡蛎MITF氨基酸序列进行了序列分析和多序列比对,分析了长牡蛎幼体发育各时期的转录组,采用荧光定量PCR的方法研究长牡蛎各组织及黑壳、白壳牡蛎特定部位mRNA表达情况。4个MITF基因中有3个基因可能为假基因,有表达的长牡蛎MITF基因共编码448个氨基酸,为亲水性不稳定蛋白,含有N端结构域(MITFTFEBC3N superfamily)和高度保守的功能性结构域HLH结构域。转录组分析发现MITF在长牡蛎个体发育的各个时期均有表达,在稚贝期达到最高。组织表达结果显示MITF在外套膜中的表达水平显著高于其他组织。MITF在黑壳牡蛎闭壳肌中的表达量显著低于白壳牡蛎,而在黑壳牡蛎外套膜中的表达量高于白壳牡蛎,但不显著。在黑壳、白壳牡蛎外套膜边缘的表达量都显著高于内侧。研究表明,长牡蛎MITF基因可能在牡蛎壳形成早期就参与了黑色素的生成调控和个体的生长发育,可调控牡蛎酪氨酸酶Tyr2基因,参与外套膜和贝壳中黑色素的形成。本研究为进一步研究牡蛎壳色形成机制奠定了基础。     

合浦珠母贝热休克蛋白hsp70基因的克隆与表达分析

采用同源克隆和RT-PCR技术对合浦珠母贝(Pinctada fucata)热休克蛋白hsp70基因进行了克隆和表达分析。获得cDNA全长序列2 365 bp,其中3’非编码区域(UTR)为318 bp,5’UTR为88 bp,开放阅读框(ORF)为1 959 bp,编码652个氨基酸,分子量约为71.39 kD,理论等电点为5.22,并含有3个HSP70家族的签名序列IDLGTTYS、DLGGGTFD和EEVD。同源性分析表明,合浦珠母贝HSP70的氨基酸序列与太平洋牡蛎(Crassostrea gigas)等双壳贝类的相似性高达86%以上,基于氨基酸序列的聚类分析表明,合浦珠母贝与牡蛎属种类亲缘关系最近。高温、高盐刺激后,半定量RT-PCR检测发现hsp70基因的表达明显增加,高温刺激的表达量高于高盐刺激,高温刺激组不同组织的表达量由大到小依次为鳃、消化腺、外套膜、肌肉、性腺,高盐刺激组不同组织的表达量由大到小依次为鳃、外套膜、肌肉、消化腺、性腺,表明HSP70参与了机体对刺激的应答过程。该基因的克隆为进一步深入研究合浦珠母贝的抗逆机理及其遗传改良奠定了重要基础。     

马氏珠母贝转录组数据的重新组装

对本实验室和其他学者已经发表的马氏珠母贝外套膜和珍珠囊的454焦磷酸测序转录组数据以及NCBI中的EST数据进行了重组装、同源序列比对和GO注释分析。重组装获得了30266个contig,平均长度522 bp,其中最长的contig为4144 bp;NR数据库BLAST分析获取了2310个相似性contig(E≤10?5),其中1902个在模式动物(人、小鼠、海胆、线虫、斑马鱼、果蝇或者太平洋牡蛎和珠母贝)中可以查找到同源序列。基因注释(GO)结果表明,注释到生物学过程的3846个contig可分为23个亚类,其中代谢过程蛋白、细胞过程蛋白和生物学调控蛋白为contig数量最多的3个亚类;注释到分子功能的4601个contig共分为11个亚类,其中结合蛋白的contig数量最多,其次是具有催化活性的蛋白和结构分子活性蛋白;注释到细胞成分的2992个contig分为17个亚类,其中细胞和细胞组成部分的contig数量最多,其次分别是细胞器和大分子复合体contig数量。本研究结果有助于开展外套膜和珍珠囊特异性表达基因的大规模筛选。