果树自交不亲和性的遗传与生理机制及其研究

在简要介绍植物（包括果树）自交不亲和性与生理控制机制的基础上，着重对梨、苹果、杏等果树自交不亲和性的研究进行了综述，内容包括花柱内花粉管生长特点、花柱S糖蛋白的特性与作用机理以及果树自交不亲和的分子生物学研究，并提出了该领域有待研究的问题。     

茉莉酸对苹果气孔运动及抗旱性的影响

50、75、100、150 mg／L JA处理苹果叶片表皮，使开放气孔的气孔开度分别减小27．4％、37．5％、56．9％和63．8％，使关闭后的气孔在光下开放时气孔开度分别减小15％、18％、28．8％、52．5％、61．3％；处理植株后干旱胁迫，叶片相对电导率分别降低15％、18％、31％、58％，脯氨酸含量分别提高0．09％、0．40％、0．42％、0．54 ng／gFW，可溶性糖含量分别提高1．89％、2．53％、3．80％、5．06％。25 mg／L JA处理与对照均无显著差别。JA的作用表现出明显的浓度效应。     

苹果梨花芽孕育期生长锥细胞超微结构的观察

利用透射电镜对苹果梨花芽孕育期芽内生长锥细胞进行了观察，结果显示，在花芽孕育形成的过程中，生长锥细胞的超微结构发生了一系列变化：蛋白体和脂类小滴逐渐增加；核外膜向外延伸到胞质中与内质网相连，其上分布的核糖体增多；线粒体和内质网数目丰富且分布均匀；核糖体大量游散在胞质中；质体呈浅裂，中央缢缩；微管分布于靠近质膜处等。以上超微结构的变化表现出细胞旺盛分裂和分化的特点，反映了花芽孕育过程中的细胞学特征。     

Effects of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus

This study was conducted to evaluate the effects of different concentrations of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus. Diets were formulated to contain 40% crude protein from solvent-extracted menhaden fish meal and 0, 7, 14 or 21% lipid from menhaden fish oil. The basal diet, without supplemental fish oil, contained lipid at 0.4% of dry weight. The diets were fed to groups of 25 juvenile red drum initially averaging 7.3 ± 0.18 g fish^–1 in a recirculating culture system for 8 weeks and weight gain was recorded. After an additional 8 weeks, 16 fish from each treatment were sacrificed and the following measurements were recorded: hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, and liver -tocopherol, malondialdehyde (MDA) formation, and cytochrome P-4501A activity (measured as 7-ethoxyresorufin O-deethylase (EROD) activity). The activity of alanine and aspartate aminotransferases and concentrations of -tocopherol also were measured in plasma.Weight gain was significantly (p<0.05) affected by dietary lipid concentration, with values ranging from 361% of initial weight for fish fed the basal diet to 527% of initial weight for fish fed the diet containing 7% lipid. The HSI and IPF ratio values also were significantly affected by lipid with the lowest values recorded for fish fed the basal diet and the highest values observed in fish fed the diet containing 21% lipid. Increasing dietary lipid significantly increased oxidative stress as reflected in reduced -tocopherol in liver and plasma and increased MDA formation in the liver, although no overt pathological signs were observed. These findings suggest that lipid concentrations between 7 and 14%, when the diet contains 60 IU vitamin E kg^–1, are likely to limit oxidative stress and result in normal physiological responses of red drum.     

玫瑰黄链霉菌Men-myco-93-63负调控基因nsdAmgh的克隆及序列分析

nsdA (Negative regulator of Streptomyces differentiation)基因是天蓝色链霉菌中发现的与抗生素合成相关的负调控基因.根据天蓝色链霉菌 A3(2)的序列设计引物扩增玫瑰黄链霉菌 Men-myco-93-63 菌株中的该基因,经测序和序列分析表明,由保守序列引物nsdA-2R和nsdA-2L克隆到的Men-myco-93-63 菌株中的该基因 800 bp片段与已知nsdA基因核苷酸保守区域序列同源性达到99%,由全长序列扩增引物nsdA WZ-R和nsdA WZ-L克隆到的该基因包含一个完整的开放阅读框,共编码 369 个氨基酸,与变铅青链霉菌和天蓝色链霉菌A3(2)中的nsdA 基因的核苷酸序列同源性达到100%,氨基酸序列同源性达到99%.该全长基因命名为nsdA_(mgh),其在玫瑰黄链霉菌 Men-myco-93-63 中的发现为阻断该负调控基因以期获得抗生素高产工程菌株提供了重要依据.     

HIV VRC01单链抗体的构建、原核表达及纯化

【目的】合成、克隆HIV单克降抗体VRC01的单链抗体基因,在大肠杆菌中诱导表达并进行纯化。【方法】根据GenBank上公布的VRC01抗体可变区的氨基酸序列合成引物,运用基因从头合成和重叠延伸PCR方法,克隆抗体VRC01的重链可变区(VH)和轻链可变区(VL)基因;用15肽Linker(Gly4Ser)3将VH和VL基因相连,得到单链抗体基因scFv-VH-Linker-VL和scFv-VL-Linker-VH,并分别将两者克隆到原核表达载体pET28a中,构建重组质粒后转入大肠杆菌BL21(DE3+)中诱导表达,对表达产物进行SDS-PAGE和Western blot分析;将重组蛋白进行变性处理后,利用Ni-NTA金属螯合层析柱对表达产物进行初步纯化。【结果】得到了VRC01的2种单链抗体的基因scFv-VH-Linker-VL和scFv-VL-Linker-VH,其大小分别为770和768bp。SDS-PAGE电泳结果显示,scFv-VH-Linker-VL在大肠杆菌中不表达或表达量极低;而scFv-VL-Linker-VH表达量较高,蛋白质分子质量约为29ku,且主要以包涵体形式存在。Western blot检测结果证实,scFv-VL-Linker-VH可与His-Tag融合表达,并通过亲和层析法成功纯化获得了该单链抗体。【结论】成功构建、表达并纯化出了VRC01单链抗体,为进一步研究VRC01单链抗体的生物学特性奠定了基础。     

森林经营方案实施效益计算机评价系统的研制与应用

本文论述“国营林场、采育场森林经营方案实施效益评价系统”的计算机程序设计思想以及程序实现的方法,介绍福建省来舟林业试验场及其森林经营方案概况,把各种量化的数据输入计算机,对该场森林经营方案实施效益进行了评价,并讨论分析了计算机输出的评价结果。     

玫瑰黄链霉菌对根结线虫的防效及对土壤微生物区系的影响

利用玫瑰黄链霉菌Men-myco-93-63 菌株的固、液体发酵物防治设施番茄和黄瓜根结线虫,研究其对根结线虫的防效以及施用生防菌对土壤微生物区系的影响。结果表明：Men-myco-93-63 菌株固体发酵物1 V∶200 V对黄瓜和番茄根结线虫病的防效分别达到了65.62% 和57.69%;施用生防菌使得植株根际土壤中细菌数量增加了19.57%,放线菌数量增加了316.67%,真菌数量降低了28.57%,以上3 种可培养微生物总量由7.4×10⁶ cfu·g^-1增加到16.75×10⁶ cfu·g^-1,增加了126.35%。     

Characterization of an Alginate Lyase,FlAlyA, from Flavobacterium sp. Strain UMI-01 and Its Expression in Escherichia coli

A major alginate lyase, FlAlyA, was purified from the periplasmic fraction of an alginate-assimilating bacterium, Flavobacterium sp. strain UMI-01. FlAlyA showed a single band of ~30 kDa on SDS-PAGE and exhibited the optimal temperature and pH at 55 °C and pH 7.7, respectively. Analyses for substrate preference and reaction products indicated that FlAlyA was an endolytic poly(mannuronate) lyase (EC 4.2.2.3). A gene fragment encoding the amino-acid sequence of 288 residues for FlAlyA was amplified by inverse PCR. The N-terminal region of 21 residues except for the initiation Met in the deduced sequence was predicted as the signal peptide and the following region of six residues was regarded as propeptide, while the C-terminal region of 260 residues was regarded as the polysaccharide-lyase-family-7-type catalytic domain. The entire coding region for FlAlyA was subjected to the pCold I—Escherichia coli BL21(DE3) expression system and ~eight times higher yield of recombinant FlAlyA (recFlAlyA) than that of native FlAlyA was achieved. The recFlAlyA recovered in the periplasmic fraction of E. coli had lost the signal peptide region along with the N-terminal 3 residues of propeptide region. This suggested that the signal peptide of FlAlyA could function in part in E. coli.     

MiRNA－93－5p 对 VEGF 的转录调控及其与梅花鹿茸细胞增殖的关系1）

为了探讨miRNA－93－5p对梅花鹿血管内皮生长因子（ VEGF）的转录调控作用及其与鹿茸细胞生长的关系,分离了鹿茸顶端软骨组织细胞,利用Trizol试剂法提取细胞总RNA,反转录合成cDNA。根据GenBank已发表的相关序列设计梅花鹿VEGF基因的3′端非编码区部分序列（3′UTR）特异引物并进行克隆,构建VEGF基因的3′UTR野生型及其突变体序列双荧光素酶报告基因载体并进行荧光素酶活性检测。再将人工合成的miRNA－93－5p模拟物转染鹿茸软骨细胞,MTT法检测鹿茸细胞体外增殖的变化;Western blotting分析VEGF蛋白的表达丰度。结果表明：成功获得了鹿茸组织VEGF基因的3′UTR序列,野生型序列长度为356 bp,突变体长度为336 bp。荧光素酶活性检测结果表明,转染野生型质粒组细胞荧光素酶活性降低,而转染突变体组细胞荧光素酶活性无明显变化。 MTT法和Western blotting结果显示,鹿茸细胞的体外增殖受到抑制,VEGF蛋白的表达水平下降,且呈时间依赖性。