对虾性别分化、性腺发育中若干关键问题的研究

利用扫描电镜和组织学研究相结合的方法研究了中国对虾早期的性别分化。扫描电镜观察的结果表明，中国对虾外部性征的性别分化是在变为仔是后第35天开始的，而内部性征则是在变为仔虾后的第76天观察到生殖腺原基向精巢和卵巢的分化。促雄腺(Androgenic gland)是控制甲壳动物性别分化的一个重要器官。到目前为止，已在软甲类的多数目中报道了促雄腺的存在。不同种类促雄腺的分布位置不同，外部形态也有很大差别。中国对虾的促雄腺位于第五对步足基部的肌肉间，于片状覆盖在精荚囊与射精管连接处。当用苏木精染色时，由于其嗜碱性比较强，整个腺体被染成深蓝色。扫描电镜观察可见，中国对虾的促雄腺具有葡萄串状的外形，它是由许多葡萄形的亚单位组成的，这些亚单位的大小、形态各异组织学观察的结果表明，中国对虾的促雄腺由两种类型的细胞组成。第一类细胞，数量少，核的嗜碱性很强，被苏木精染成深蓝色，核仁不易分辨；第二类细胞，数量大，核比较膨大，嗜碱性较弱，核质疏松，具有明显可见的核仁。不同大小的虾中，腺细胞的排列方式不同。随着对虾的发育成熟，腺细胞逐渐由索状排列变为成团排列，说明促雄腺的形成存在一个逐渐发育的过程。利用手术破坏促雄腺或用促雄腺匀浆液浸泡仔虾的方法研究了中国对虾促雄腺的功能，中国对虾促雄腺的存在与否对于雄性外部性征的发育和维持起着重要作用。破坏促雄腺的雄虾的交接器的发育明显受到抑制。但是，未观察到促雄腺对对虾内部性征发育的影响，这可能与进行手术时对虾所处的发育期有关。利用促雄腺的匀浆液浸泡仔虾，对雄虾交接器的发育有一定促进作用。高温处理对中国对虾受精卵孵化率的影响随处理温度及处理时间的改变而发生变化。用热处理方法成功地诱导了中国对虾三倍体，最高诱导率达75%，并对三倍体群体的生物学特性进行了研究。在中国对虾三倍体的群体中，雌体的巢存在两种类型：一种类型卵巢，生殖细胞停留在卵原细胞阶段，没有进一步发育；另一种类型的卵巢，生殖细胞由卵原细胞发育为初级卵母细胞，发育情况与对虾的类似。对卵巢处于发育的第II或第III期的中国对虾以及卵巢发育处在第I、II、III期的几纳对虾的血液和卵巢中法呢烯酸甲酯（MF）含量进行了研究，结果表明，中国对虾和凡纳对虾中均存在MF。凡纳对虾血液和卵巢发育具有明显的刺激作用，这种刺激作用是通过血液中法呢烯酸甲酯含量的提高来实现的。这一研究结果进一步说明了法呢烯酸甲酯与甲壳动物繁殖之间的关系。

Toxin-screening and identification of bacteria isolated from highly toxic marine gastropod Nassarius semiplicatus

Bacteria isolated from a highly toxic sample of gastropod Nassarius semiplicatus in Lianyungang, Jiangsu Province in July 2007, were studied to probe into the relationship between bacteria and toxicity of nassariid gastropod. The toxicity of the gastropod sample was 2 x 10(2) mouse unit (MU) Per gram Of tissue (wet weight). High concentration of tetrodotoxin (TTX) and its analogues (TTXs) were found in the digestive gland and muscle of the gastropod, using high performance liquid chromatography coupled with mass chromatography (LC-MS). Bacterial strains isolated from the digestive gland were cultured and screened for TTX with a competitive ELISA method. Tetrodotoxin was detected in a proportion of bacterial strains, but the toxin content was low. Partial 16S ribosomal DNA (rDNA) of the TTX-producing strains was then sequenced and compared with those published in the GenBank to tentatively identify the toxic strains. It was found that most of the toxic strains were closely affiliated with genus Vibrio, and the others were related to genus Shewanella, Marinomonas, Tenacibaculum and Aeromonas. These findings suggest that tetrodotoxin-producing bacteria might play an important role in tetrodotoxin accumulation/production in N. semiplicatus. (C) 2008 Elsevier Ltd. All rights reserved.

Identification of quantitative trait loci for growth-related traits in the Pacific abalone Haliotis discus hannai Ino

The locations and effects of quantitative trait loci (QTL) were estimated for nine characters for growth-related traits in the Pacific abalone (Haliotis discus hannai Ino) using a randomly amplified polymorphic DNA (RAPD), amplification fragment length polymorphism (AFLP) and SSR genetic linkage map. Twenty-eight putatively significant QTLs (LOD > 2.4) were detected for nine traits (shell length, shell width, total weight, shell weight, weight of soft part, muscle weight, gonad and digestive gland weight, mantle weight and gill weight). The percentage of phenotypic variation explained by a single QTL ranged from 8.0% to 35.9%. The significant correlations (P < 0.001) were found among all the growth-related traits, and Pearson's correlation coefficients were more than 0.81. For the female map, the QTL for growth were concentrated on groups 1 and 4 linkage maps. On the male map, the QTL that influenced growth-related traits gathered on the groups 1 and 9 linkage maps. Genetic linkage map construction and QTL analysis for growth-related traits are the basis for the marker-assisted selection and will eventually improve production and quality of the Pacific abalone.

A preliminary analysis of the phylogeny of the Swertiinae (Gentianaceae) based on ITS data

To better understand the floral diversity and the phylogeny of the Swertiinae (Gentianaceae), the internal transcribed spacer (ITS) region of nrDNA for 17 species and one outgroup was sequenced. Our data suggest that corolla type, gland shape, and corolla appendage are poorly correlated with the ITS phylogeny. The genus Swertia s.l. and the rotate group previously recognized based on the corolla types and gland shapes are polyphyletic. Four genera with simple protruding glands and three taxa with corolla appendages are not clustered as the monophyletic groups. Four separate clades, corresponding to the four sections, were identified in Swertia s.1. Lomatogoniopsis with the simple protruding gland type of the tubular group closely related to Lomatogonium of the rotate group. The deeply lobing corolla and concave foveae may be ancestral in the Swertiinae, while the tubular corolla and the protruding glands may have undergone convergent evolution.

生物钟基因mPer2调控节律性免疫功能的机制研究

This project was carried out with the aims to investigate the mechanism of circadian immune regulation by one of the core Clock gene, mPer2. To achieve this, we selected mPer2 knock out (mPer2-/-) mice as the optimal animal model. Two different approaches were performed. In the first approach, we injected WT or mPer2-/- mice with an equal dosage of lipopolysaccharide (LPS), and systematically measured serum corticosterone induction, expression of core Clock genes, as well as a key enzyme for corticosterone metabolism (mStAR) in adrenal gland. We found that the acute induction of corticosterone and mStAR were closely associated with the circadian immune response to LPS. Besides, real time quantitative PCR (q-PCR) and luciferase assay consistently showed that mStAR is a Clock controlled gene in adrenal gland, where its expression is negatively influenced by mPer2. In the second approach, expression level and circadian manner of 11 cytotoxicity regulation genes in WT or mPer2-/- mice bone marrow were measured by q-PCR in order to explore the candidate genes which could mediate the circadian immune regulation by mPer2. We found that expression level of Ly49C, Ly49I, and Nkg2d was significant down-regulated in mPer2-/- mice. Further, we found that daily expression of Ly49C and Nkg2d fluctuated in a circadian manner in WT mice, where these rhythms were disrupted in mPer2-/- mice. Thus, it was suggested that these two cytotoxic genes were two clock controlled genes whose circadian expression were regulated by mPer2. Taken together, our results suggested that corticosterone, mStAR, Ly49C, and Nkg2d were four candidate molecules that may mediate the circadian immune response regulation by mPer2.