清初苏州园林涧上草堂钩沉

涧上草堂是明末清初苏州地区由明代遗民徐枋所构建的古典园林。明清易代之际,很多明朝文人士大夫不愿出仕新朝并自愿成为明遗民,成为清初一大特殊社会现象,对这一特殊群体所构建的园林进行研究,可以在一定程度上丰富中国的古典园林史。通过分析画作与历史文献,对涧上草堂的原址、各园林要素以及园林后期的发展3个方面进行研究,得出草堂的平面想象图并概括涧上草堂的历史价值,力图推进在明末清初这一过渡时间段古典园林研究与发展的深度。     

彩鲫与建鲤杂交F1肠道黏膜显微观察

通过对全红体色彩鲫(Carassius auratuas)与建鲤(Cyprinus carpio var.Jian)的正反交杂交后代进行消化道生长发育的比较研究,以期为建鲤与彩鲫的远缘杂交生产和研究提供理论依据。本实验选择建鲤亲本、彩鲫亲本、鲤鲫F1(建鲤♀×彩鲫♂)和鲫鲤F1(彩鲫♀×建鲤♂)各50条,每组实验鱼日龄相近。每组选5条体重相近的鱼测量肠长和肠重,并制作肠切片,计算肠重指数、肠长指数、肠道皱襞高度、绒毛长、绒毛宽、肌层厚度数据。数据结果采用SPSS(17.0)软件中的LSD法进行显著性检验。结果表明,以彩鲫作为父本的F1的消化水平高于以彩鲫作为母本的F1,且彩鲫作为父本的F1即鲤鲫F1的消化水平高于两亲本,体现出了杂种优势。     

鲤和鲫线粒体(mtDNA)全基因组分析

为研究鲤(Cyprinus carpio)和鲫(Carassius auratus) mtDNA上的基因分布特征,丰富鱼类mtDNA数据库。本研究通过PCR扩增和测序,获得了鲤和鲫mtDNA基因组序列,经比对分析表明,鲤和鲫的mtDNA序列全长均为16 596 bp,共有13个蛋白质编码基因、22个tRNA基因、2个rRNA基因和1个D-Loop区。鲤和鲫的mtDNA序列中(A+T)百分含量分别为57.2%和57.1%,其碱基组成均具有一定的A/T碱基偏向性。N-J系统发育树分析表明,鲤和鲫与琵琶湖鮈亲缘关系最近,与达氏深水尾魟亲缘关系最远。本研究揭示了鲤和鲫mtDNA遗传变异特征及基因分布规律,为鱼类遗传资源保护及开发利用提供了参考依据。     

草鱼EST-SSR标记及5个不同地域群体的遗传结构分析

以草鱼(Ctenopharyngodon idella)脑、肌肉、肝等组织构建cDNA文库,经测序获得unigenes序列45 318个,从中筛选微卫星序列共5 556个,据此设计EST-SSR引物118对,其中19对引物能够扩增出带型清晰且多态性较高的谱带。用这19个EST-SSR标记研究3个长江水系群体(石首、监利和长沙)和2个珠江水系群体(清远和肇庆)草鱼的遗传结构。结果表明,5个群体的平均多态信息含量(PIC)为0.415 4～0.460 4,显示草鱼群体的遗传多样性偏低;平均观测杂合度(Ho)为0.415 8～0.501 3,平均期望杂合度为(He)0.450 6～0.502 8,其中,长沙群体平均期望杂合度最高,为0.502 8,监利群体的平均观察杂合度和平均期望杂合度均最低,分别为0.415 8和0.450 6,即长沙群体的遗传多样性最高,监利群体的遗传多样性最低;对数据进行F-检验,结果表明,群体间的遗传分化程度低。Hardy-Weinberg平衡的卡方检验结果表明,5个群体均一定程度上偏离了平衡;聚类分析显示长沙群体、石首群体与监利群体聚成一支;肇庆群体与清远群体聚成一支,这与草鱼群体的流域分布一致。     

洞庭湖水系沅水和澧水野鲫的染色体组型及资源保护

鲫(Carassius auratus)是洞庭湖水系一种重要的经济鱼类。为了解洞庭湖水系野鲫的细胞遗传背景,采用PHA和秋水仙素活体注射法,对沅水和澧水采集的野鲫样本逐一进行肾细胞染色体制片及组型分析。结果发现,在两条河流的野鲫群体中均检测出染色体数为100和基本染色体数为150的两种不同倍性个体,其中,两条河流染色体数为100的二倍体鲫组型公式为2N=28M+22SM+28ST+22T,NF=150;基本染色体数为150的三倍体鲫组型公式为3N=42M+33SM+42ST+33T,NF=225。在沅水和澧水不同采样点随机采集的共100尾野鲫中,检测出的三倍体比例(85%)远高于二倍体(15%),且二倍体与三倍体鲫个体在形态特征上不存在明显差异(P0.05)。两种不同倍性鲫在同一水体的共存对于鲫的遗传进化与选育具有一定的理论和实践意义,而二倍体鲫种群的大量减少,则提示我们应该从染色体组遗传多样性角度加强对洞庭湖水系二倍体野鲫资源的保护。     

Stimulation of antioxidant enzymes levels in carp (Cyprinus carpio L.) infected by Ptychobothrium sp. (Cestoda)

Increased antioxidant enzymatic activities were observed in carp parasitised by Ptychobothrium sp. when compared with healthy fish. This antioxidant response could contribute to neutralise the oxidative stress normally induced by parasitism.     

Autoradiographic distribution of neuropeptide Y binding sites in the brain of the carp Cyprinus carpio L. (Cyprinidae,Teleostei)

The present study reports the distribution of neuropeptide Y (NPY)-binding sites in the brain of the adult carp Cyprinus carpio L. Radioiodinated NPY was used as tracer in the autoradiographic procedure. The NPY-binding sites (NPY-bs) were widely distributed in the carp brain. Generally, a good match was observed between the distribution of NPY-bs and the distribution of NPY-immunoreactive (NPY-ir) elements previously reported in the forebrain of the carp. Low to moderate concentration of NPY-bs were found in the telencephalon, this finding indicates that NPY may play a role in the processing of olfactory inputs and in more complex behaviours like spatial learning acquisition and retention, whose importance could correlated with similar results obtained in mammals. Moreover, in the rhombencephalon, the presence of NPY-bs at level of lobus vagus and the lobus facialis suggests that NPY may be implicated in food-seeking behaviour and swallowing reflex.     

Glycosylation is essential for translocation of carp retinol-binding protein across the endoplasmic reticulum membrane

Retinoid transport is well characterized in many vertebrates, while it is still largely unexplored in fish. To study the transport and utilization of vitamin A in these organisms, we have isolated from a carp liver cDNA library retinol-binding protein, its plasma carrier. The primary structure of carp retinol-binding protein is very conserved, but presents unique features compared to those of the correspondent proteins isolated and characterized so far in other species: it has an uncleavable signal peptide and two N-glycosylation sites in the NH(2)-terminal region of the protein that are glycosylated in vivo. In this paper, we have investigated the function of the carbohydrate chains, by constructing three mutants deprived of the first, the second or both carbohydrates. The results of transient transfection of wild type and mutant retinol-binding protein in Cos cells followed by Western blotting and immunofluorescence analysis have shown that the absence of both carbohydrate moieties blocks secretion, while the presence of one carbohydrate group leads to an inefficient secretion. Experiments of carp RBP mRNA in vitro translation in a reticulocyte cell-free system in the presence of microsomes have demonstrated that N-glycosylation is necessary for efficient translocation across the endoplasmic reticulum membranes. Moreover, when Cos cells were transiently transfected with wild type and mutant retinol-binding protein (aa 1-67)-green fluorescent protein fusion constructs and semi-permeabilized with streptolysin O, immunofluorescence analysis with anti-green fluorescent protein antibody revealed that the double mutant is exposed to the cytosol, thus confirming the importance of glycan moieties in the translocation process.     

Gene structure of the carp fish ribosomal protein L41: seasonally regulated expression

The seasonal acclimatization of the carp fish demands physiological compensatory responses. The process involves profound nucleolar adjustments and remarkable changes in rRNA synthesis, which affect ribosomal biogenesis. We have documented that protein kinase CK2, whose activity is related to ribosomal protein L41 and the regulation of rRNA synthesis, was expressed in notably higher amounts in summer-acclimatized carp compared to the cold-season adapted fish. Thus, we approached the study of the functional genomics of carp L41 protein. We report the first cloning of a fish L41 gene encoding the highly conserved 25 amino acids, including approximately 1700 bp regulatory upstream region and the 3(') polyadenylation signal, plus the isolation and characterization of two different L41 cDNAs. We found a clear differential expression of L41, which follows the same pattern as protein kinase CK2beta that transcribes at higher levels in the summer-acclimatized carp than it does in the winter-adapted fish.