栗属分子生物学研究进展

壳斗科(Fagaceae)栗属(Castanea)植物的7个种分布广泛,不仅可用于木材生产,而且在坚果生产上也占有独特地位。基于形态学、同工酶、子叶储藏蛋白和RAPD数据,通过对分布于亚洲、欧洲和北美的栗属植物的遗传多样性研究表明中国板栗(Castaneamollissima)是世界栗属植物的原始种,长江流域是中国板栗的遗传多样性中心,土耳其是欧洲板栗的起源中心之一。在分子标记辅助育种方面,已发表了基于形态学、同工酶、RAPD和ISSR数据的两张栗属植物的遗传连锁图谱,其中一个是用欧洲板栗种内杂交后代F1全部单株构建的,另一张是用美洲板栗与中国板栗种间杂交后代F2单株构建的,并已经定位了3个假定的栗疫病抗性位点;并且证明在多年生木本果树上同工酶基因可通过连锁关系分析与形态基因整合为1个单一基因图而无需另外的杂交。从栗属植物中已分离纯化了包括可抑制HIV-1反转录酶活性的Mollisin在内的几丁质酶等抗真菌蛋白、胱氨酸蛋白酶抑制剂、热击蛋白、红血球凝集素、脱水素、花粉过敏原等功能蛋白质,并已经克隆了包括伤害应答基因在内的部分功能蛋白质相关基因。因此,作为具有独特性质的栗属植物,有必要开展更多的研究,就此本文对栗属植物遗传多样性、分子标记辅助育种和功能蛋白纯化和有关基因克隆等几方面的研究进展进行了总结,希望能位同行提供参考。     

植物热激转录因子在非生物逆境中的作用

非生物逆境通常导致生物体内蛋白变性。热激蛋白(Hsp)作为分子伴侣协助蛋白的重新折叠、稳定、胞内运输和降解,以阻止受损蛋白的累积,维护细胞内环境的稳定。而热激蛋白的表达是通过热激转录因子(Hsfs)结合于热激蛋白基因的启动子的热激元件上(heatshockelement,HSE),以募集其它转录因子而形成转录复合体,促进热激蛋白基因的表达。植物热激转录因子比动物系统更为多样性。根据其基本的结构域,植物热激转录因子可分为三类:HsfA、HsfB、HsfC。A类Hsfs已有大量深入的研究和报道,特别是在番茄方面。HsfB和HsfC的作用尚不清楚。在其复杂的网络中,每一热激转录因子均有其独特的作用,取决于其表达模式、亚细胞定位、聚合化、活性及与其他蛋白的相互作用。在非生物逆境,尤其是热激逆境下,A类热激转录因子在调节热激蛋白的表达起着重要作用。番茄的HsfA1起着主导作用,其缺失无法被其他相近的Hsfs所取代,但在持续热逆境下,在HsfA1的配合下,HsfA2可成为主要调节因子。B类热激转录因子可作为A类Hsfs的阻抑蛋白。然而,基于对不同的单个突变体的研究,以及对酵母Hsf1致死突变体的拯救恢复,一些热激转录因子的作用又是丰余的。此外,热激蛋白也对热激转录因子起负反馈调节作用。     

农杆菌介导法将Bt基因转入大豆的研究

以6个品种大豆体细胞胚为受体,以Bt基因为目的基因,应用根癌农杆茵介导法对大豆进行了遗传转化,经体细胞胚萌发、生根及壮苗培养和卡那霉素选择培养,获得了完整的抗性再生苗,经PCR、PCR-Southern和点杂交分析鉴定,初步证明外源基因Bt已整合到大豆的基因组中。     

Vitellogenin in tilapia (Oreochromis mossambicus): Induction of two forms by estradiol,quantification in plasma and characterization in oocyte extract

Two forms of vitellogenin were isolated by DEAE agarose ion-exchange chromatography from plasma of the tilapia, Oreochromis mossambicus. The monomers have apparent molecular masses of 200 and 130 kDa, as indicated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and a total amount of phosphorus of 1.7 and 0.1%, respectively. Antibodies specific to the two forms, designated tVTG-200 and tVTG-130, were generated in rabbits and used to develop enzyme-linked immunosorbent assays (ELISAs) and in Western blot analyses of plasma and oocyte extract. SDS-PAGE of the oocyte extract showed a major protein band at 106.6, minor bands at 26.6, 24.2, and 23.7 kDa, and very faint bands at 83.4 and 17.5 kDa. Western blots of the oocyte extract revealed that the antiserum to tVTG-200 recognized strongly the protein bands at 24.2 and 23.7 kDa, and less strongly the bands at 25.1 and 22.6 kDa, whereas the antiserum to tVTG-130 recognized mainly the protein band at 106.6 kDa. The presence of both VTGs in untreated male tilapia was detected with the ELISAs using relatively high plasma volumes. Their presence in males was confirmed by VTG-like immunoreactive materials eluting from the ion-exchange column at the same positions as tVTG-200 and tVTG-130. The concentrations of the VTGs in males were several orders of magnitude lower than in vitellogenic females. Treatment of male tilapia with estradiol-17β (E₂) induced both VTGs within 24h. After 7 days, tVTG-130 reached a maximum concentration in plasma, whereas tVTG-200 continued to increase. Our findings demonstrate that the two vitellogenins are biochemically distinct, possibly differentially regulated, and made by both sexes.     

Intra- and interspecific phenotypic characteristics of fish-pathogenic Edwardsiella ictaluri and E. tarda

Intra‐ and interspecific characteristics of fish‐pathogenic Edwardsiella ictaluri, and E. tarda were determined by numerical analysis of gel electrophoresed protein profiles, fatty acid methyl esters (FAMEs) and immunoblotting. The 18 E. ictaluri isolates revealed a high degree of homogeneity (70% similarity or higher) in their protein profiles and 95% similarity in their FAME, while the nine E. tarda isolates revealed 30% similarity in their protein profiles and 95% similarity in their FAME. Immunoblots probed for antigenic epitopes with goat antiserum produced against E. ictaluri and E. tarda, respectively, revealed that E. ictaluri were more homogeneous compared with the E. tarda isolates. Overall, there was a considerable degree of relatedness between the two species. Our findings suggest that phenotypically E. ictaluri represents a clonal bacterial population structure compared with the less monomorphic E. tarda.     

Apparent digestible protein,energy and amino acid availability of three plant proteins in Florida pompano,Trachinotus carolinus L. in seawater and low‐salinity water

Two experiments were conducted with Florida pompano, Trachinotus carolinus L. at 3 and 28 g L^?1 salinity to determine apparent crude protein digestibility (ACPD), energy digestibility (AED) and amino acid availability (AAAA) from soybean meal (SBM), soy protein isolate (SPI) and corn gluten meal (CGM). Mean AAAA was similar to ACPD. In fish adapted to 3 g L^?1 salinity, they were 81.2% and 81.9% (CGM), 93.6% and 92.2% (SBM), 93.8% and 93.1% (SPI) for AAAA and ACPD respectively. In fish adapted to 28 g L^?1, they were 84.5% and 83.4% (CGM), 86.5% and 87.1% (SBM), and 83.4% and 85.0% (SPI) for AAAA and ACPD respectively. The AED was highest for SPI and lowest for SBM and inversely related to carbohydrate. The ACPD, AED and AAAA of soy products appeared to be lower in high salinity, whereas CGM was unaffected. The data suggest that SBM, SPI and CGM should be further evaluated as partial fishmeal replacements in Florida pompano diets. Application of the generated coefficients can be used to develop well‐balanced, low‐cost diets for Florida pompano reared in low salinity or seawater.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.     

转Bt-基因棉棉仁粉对鲤鱼营养价值的研究

为评估转Ｂｔ－基因棉棉仁粉用作鱼饲料的有效性和安全性,将其配成试验饲料进行了１４周鲤鱼喂养,前８周着重就鲤钱垢增重率、特定生长率、铒料系数、鱼体化学成分含量等营养学指标进行了观察。上述各项指标经统计分析,在试验组和对照组之间均无显著性差异。表明转Ｂｔ－基因棉棉仁粉经脱棉酚处理后,与常规棉棉仁粉一样可作鱼用饲料的蛋白源。     

纳米铜晶胞体积弹性模量变化规律的分子动力学模拟

用分子动力学的方法详细模拟纳米铜晶胞随温度与压强变化的规律,得到了铜晶胞体积弹性模量突变的敏感压强点.模拟结果表明:单晶铜的在压强小于75GPa时体积弹性模量随温度升高而降低,随压强增大而增大;在压强大于75GPa时,体积弹性模量随温度升高而增大,随压强增大而减小.