球状棕囊藻的形态结构、生活史及生态意义研究进展

球状棕囊藻生活史独特,在两种完全不同的生活状态--游离单细胞和粘液质的囊体之间相互转换.生活史中共出现4 种不同类型的单细胞,存在有性和无性两种生殖方式.不同的细胞能够适应不同的环境,占据不同的生态位,有利于棕囊藻细胞的生长.球状棕囊藻的囊体直径最大可达几个厘米,外被坚韧、紧密并且具有弹性,为内部的单细胞提供了一个良好的栖息和保护场所,并且储存着能量可供细胞在营养盐限制条件下继续生长.由于囊体粒径较大,浮游动物难以摄食,并且有效的抵御了细菌和病毒的侵蚀,从而降低了棕囊藻的死亡率.与其它浮游植物相比,球状棕囊藻的竞争策略更加优越,提高了球状棕囊藻在海洋生态系统中的竞争能力.     

小鼠精囊自身抗原的原核表达和纯化

目的：在原核细胞中表达小鼠精囊自身抗原（SVA）,并对表达产物进行鉴定和纯化。方法：提取小鼠附睾组织总RNA,RT-PCR获得SVA的cDNA,设计并合成特异引物序列,进一步扩增出不含信号肽的SVA编码序列,连入原核表达载体pET28a中,经酶切和测序鉴定正确的重组质粒转化大肠杆菌Rosetta（DE3）感受态细胞,IPTG诱导表达,Western印迹分析表达产物His-SVA,采用Ni-NTA纯化融合蛋白His-SVA。结果：原核表达获得融合6个组氨酸的SVA,用抗His单克隆抗体进行Western印迹鉴定,检测到相对分子质量约18×10^3的目的蛋白,与理论值一致;经Ni-NTA纯化获得较高纯度的His-SVA融合蛋白。结论：获得了在大肠杆菌中表达的小鼠附睾蛋白SVA,为后续研究其对小鼠生殖的影响奠定了基础。     

格氏栲林优势种竞争关系及其预测动态的研究

通过野外调查数据,利用Hegyi单木竞争指数模型,定量地分析格氏栲天然林种内和种间竞争强度.结果表明格氏栲种内竞争强度随着胸径的增大而逐渐减少;种内与种间竞争强度的顺序为马尾松-格氏栲(Pinus masoniana-Castanopsiskawakamii)>格氏栲-格氏栲(C. kawakamii-C. kawakamii)>木荷-格氏栲(Schima superba- C.kawakamii)＞杜英-格氏栲(Elaeocarpus decipiens-C.kawakamii)>木姜子-格氏栲(Litsea mollifolia-C. kawakamii)>老鼠矢-格氏栲(Symplocos stellaris-C.kawakamii);种内、种间竞争强度与格氏栲胸径之间存在显著的双曲线非线性回归关系,并利用模型预测了格氏栲种内种间的竞争强度.种内与种间竞争关系的数量研究,不仅拓展格氏栲天然林物种竞争规律的探索,而且为格氏栲林经营管理、保护和合理开发利用提供依据.     

沙区植物种的一类种间竞争模型及应用

以沙坡头人工固沙植物油蒿和柠条为研究对象,针对水分是沙坡头植物生r长的限制因子的现实,研究了在水分条件限制下两种植物的种间关系,建立了混播油蒿、柠条种间竞争模型,验证了油蒿、柠条既竞争又稳定共存的关系,其目的为充分有效利用水资源、稳定人工生态系统提供科学依据。     

哺乳动物精子中的ZP3结合蛋白研究进展

哺乳动物卵透明带糖蛋白ZP3(zona pellucida3)是介导精卵初级结合、诱发精子发生顶体反应的关键分子。目前已在精子中发现多种ZP3结合蛋白。95kD酪氨酸激酶受体可能通过其酪氨酸激酶活性介导ZP3诱发的顶体反应。β—1,4—半乳糖基转移酶与ZP3的糖基结合后,通过激活下游信号分子诱发顶体反应。精子蛋白sp56可能介导了顶体反应期间顶体基质与ZP之间的相互作用。透明带粘附素(zonadhesin)也是在顶体反应发生之后才与ZP发生相互作用。这些精子蛋白介导的下游信号事件将是下一步研究的热点。     

磷限制下大型海灌与微灌间资源竞争理论的实验验证

用一次性培养结合Monod方程测得海洋微藻－亚心型扁藻（Tetraselmis subcordiformis(Wile)Hazen)与大型海藻－孔石莼（Ulva pertusa Kjellm.)磷限制下的生长动力参数。孔石莼具有较低的半饱和生长常数及最大生长率,其分别为0．016μmol/L和0．16d^-1,而亚心型扁藻的半饱和生长常数和最大生长率分别是0．021μmol/L和0.83d^-1。两种藻类间的营养竞争实验采用半连续培养法在磷限制制条件下进行,实验过程中,分别对两者施予相同或不同的去除率,使两者享有相同或不同的资源需求值R^*。由Monod方程所作的竞争预测与实验观察结果的比较显示：仅在两种藻类间的资源需求值R^*差异显著（t检验,P＜0．01）时,Monod方程才能对竞争结果作出较为准确的预测;在两种藻类享的相同的资源需求值R^*时,亚心型扁藻在竞争中取代孔石莼。Monod模型仅能部分预测大型海藻与海洋微藻间的竞争结果。     

pCR3．1—bvLDH-C4′的构建及在体内外的表达

精子特异性乳酸脱氢酶(LDH—C4)在精子的运动和存活等生理活动中起着重要的作用。研究表明,LDH—C4接种鼠、兔等能够降低动物的生育率。通过RT—PER方法从布氏田鼠睾丸总RNA中克隆出编码抗原决定簇的LDH—C4基因片断;采用T—A克隆的方法将其插入到pCR3．1载体中构建重组载体pCR3．1—bvLDH—C4′,测序结果表明克隆的基因片断与已知小鼠相应片断有83％的序列同源。通过质脂体法转染HeLa细胞,RT—PCR证实其可在mRNA水平有效表达;通过肌肉接种BAIB/c小鼠,RT-PER结果表明其在体内也可有效表达。     

西双版纳聚果榕榕果小蜂种间联结性研究

应用方差比率法、2× 2列联表的 χ2 检验、联结系数、点相关系数、共同出现百分率、Pearson相关系数和秩相关系数等多指数比较的方法研究了西双版纳聚果榕榕果小蜂的种间联结性。研究表明 ,西双版纳聚果榕榕果小蜂种间总体联结性表现为极显著。 2× 2列联表的 χ2检验不能反映所研究昆虫种间联结性的真实情况。 6个聚果榕榕果小蜂种间共同出现率普遍较高。小蜂种间的联结程度多属极显著 (对于联结系数 15个种对中有 10个 ,对于秩相关系数15个种对中有 12个 )。传粉者Ceratosolenfusciceps同其它小蜂间的联结显著性最高 ,其中与Platyneuratestacea和P .mayri呈极显著负联结 ,与P .agraensis呈极显著正联结 ,但与Apoc rypta属 2小蜂的关系尚不十分明晰。Platyneura属 3种小蜂彼此之间呈极显著或显著的负联结关系 ,而Apocrypta属内 2个种之间呈极显著正联结。文章对该 6种榕果小蜂种间连接关系形成的原因及进化生态学后果作了较详尽的讨论。     

绵羊胞内单精子注射技术

In this study, the possibility of sheep transgenesis by intracytoplasmic sperm injection (ICSI) was assessed. In experiment 1, activation of ovine oocytes matured in vitro in preparation for ICSI has been investigated with 3.42 mmol/L Ca2+ treatment, ionomycin alone and ionomycin followed by 6-dimethylaminopurine (DMAP) after 3-h delay (group 1, 2 and 3, respectively). After activation, the oocytes were then cultured in SOFaaBSA medium. Cleavage rates were significantly (P<0.05) different among three groups (18.4%, 91.8% and 71.7%, respectively). In additional culture, no parthenotes in group 1, whereas 11% and 17.4% in group 2 and 3 developed to the blastocyst stage. Therefore we used the third activation method in the following ICSI tests. In experiment 2, development of ovine oocytes after ICSI was investigated. Thawed semen from two rams was separated by Percoll centrifugation and was used for ICSI or in vitro fertilization (IVF) trails. A total of 71.8% of oocytes reached the 2-cell stage following living sperm injection, which was significantly (p>0.05) different from those following IVF (41.4%) and sham-ICSI (30.2%). After seven days' culture, no sham-injected oocytes developed into the blastocyst stage, although 7% in ICSI and 16.1% in IVF-oocytes developed into the blastocyst stage, but there was no significant difference in ICSI and IVF groups (p>0.05). In the further study, the possibility of sheep transgenesis by ICSI was assessed. After coinjection of ovine oocytes matured in vitro with dead sperm cold to -20 degrees C and exogenous DNA encoding green fluorescent protein (GFP), seventy-three percent of coinjected oocytes developed to 2-cell stage (33/45) and two of them were transgene-expressing embryos. Among ten embryos at the 16-cell stage, all embryonic cells in one transgenic embryo still expressed GFP. Four coinjected blastocysts were thawed and transferred to the uterine of the two progesterone-synchronized recipient ewe. No pregnancies were detected on the 60th day. These results suggested sheep transgenic embryos could be produced by ICSI and further studies should be performed.     

蟋蟀精子表面LCA及ConA结合糖复合物的分布变化

LCA and ConA-binding glycoconjugates on cricket (Teleogryllus emma) sperm surface were detected with fluorescence microscope after FITC labelling for better understanding of the distribution of glycoconjugates during spermatogenesis and spermiogenesis. FITC-LCA and FITC-ConA were bound on the spermatocytes, and their distribution changes in the process of spermiogenesis were observed .In the testis sperm, FITC-LCA and FITC-ConA were mainly bound on the head and neck region. That is different from the mark pattern of spermatophore sperm, in which the nucleus, neck region and front of the tail showed obvious fluorescence mark, especially the acrosome complex and neck region exhibited stronger mark. The mark patterns of FITC-LCA and FITC-ConA were similar,though the former was distinctly clearer than the latter. But a little difference still exists in both of them. For example in the ninth stage of spermatid, FITC-LCA mark is located on the spermatid head and neck region, and FITC-ConA mark on the spermatid head, neck and front of the tail region. When fixed germ cells were treated with PBS instead of lectin solution, or fixed cells were incubated with lectin solution, which have been treated with 0.1 mol/L specific sugar inhibitor, i.e.α-D-mannose for FITC-LCA and FITC-ConA, and α-D-glucose for FITC-ConA, no mark was observed on the cells. Those results indicate that FITC-LCA conjugated glycoconjugates has the α-D- mannose residue, and FITC-ConA conjugated glycoconjugates has the α-D-mannose and α-D-glucose residue. The investigations show that the changes in glycoconjugates distribution of cricket sperm is similar to those of other insects and mammals. The evidence exhibit that a common rule of the glycoconjugates distribution on the sperm surface is followed by most of animal sperm which may relate to the function of sperm physiology.