见血封喉乳汁中具有细胞毒活性的强心苷

从见血封喉(Antiaris toxicaria (Pers.) Lesch.)乳汁分离得到4个强心苷类化合物,经波谱数据分析,分别鉴定为：毒毛旋花子爪哇糖苷(1)、铃兰毒苷(2)、毒毛旋花子阿洛糖苷(3)和glucostrophanthidin (4)。化合物4为首次从见血封喉中分离得到。细胞毒活性测定结果表明,化合物1~4对慢性髓原白血病细胞(K562)、人胃癌细胞(SGC-7901)和人肝癌细胞(SMMC-7721)的增殖均显示了较强的生长抑制活性。     

一株纤维素降解细菌的筛选、鉴定及产酶条件分析

目的筛选高活性的纤维素降解细菌,并进行初步鉴定和产纤维素酶条件分析。方法采集吉首旗帜山松树林的土壤样品,通过富集培养和刚果红平板染色法筛选分离纤维素降解细菌;通过形态观察、生理生化特性检测和基于16S rRNA基因序列的系统发育分析对分离的菌株进行初步鉴定。利用单因素实验对产纤维素酶条件进行优化。结果分离获得1株高活性纤维素降解细菌JDM11,初步鉴定其为Bacillus velezensis;菌株JMD11产纤维素酶最佳培养温度、最适初始pH和培养时间分别为28℃、7.0～7.5和32h,在该条件下其滤纸酶(FPase)和羧甲基纤维素酶(CMCase)活力分别为260.32U/ml和651.75U/ml。结论菌株JDM11是1株高活性纤维素降解的Bacillus velezensis。     

Cloning and Expression of a Chitinase Gene from Sanguibacter sp. C4

The chitinase Chi58 is an extracellular chitinase produced by Sanguibacter sp.strain C4. The gene-specific PCR primers were used to detect the presence of the chiA gene in strain C4. A chiA fragment (chiA-F) was amplified from the C4 genomic DNA and was used to blast-search the related sequences from the GenBank dadabase. By alignment and selection of the highly conserved regions of the homologous sequences, two pairs of primers were designed to amplify the open reading frame (ORF) of the chitinase from strain C4 by nested PCR. The results revealed that the Chi58 ORF consisted of 1 692 nucleotides encoding a protein of 563 amino acid residues. The molecular weight of the mature protein was predicted to be 58.544 kDa. The Chi58 ORF was a modular enzyme composed of a signal peptide sequence, a polycystic kidney disease I domain, and a glycosyl hydrolase family 18 domain. The chitinase of C4 exhibited a high level of similarity to the chitinase A of Serratia (88.9%-99.6%) at the amino acid sequence level. The Chi58 gene was cloned into the expression vector pET32a to construct the recombinant plasmid pChi58 and was expressed in E. coli BL-21 (DE3) cells with IPTG induction. The molecular weight of the Trx-Chi58 fusion protein was estimated to be 81.1 kDa by SDS-PAGE.     

五龙鹅MHC ClassⅠ基因克隆及同源建模研究

主要组织相容性复合体(MHC)与动物机体对外源性抗原的免疫应答之间存在关联。从GenBank/DDBJ/EMBL基因库中读取鸡、其他鸟类、爬行类和哺乳类的MHC ClassⅠ基因进行序列分析设计引物, 使用LA-PCR法从五龙鹅的基因组中克隆了MHC ClassⅠ基因序列(DNA序列和mRNA序列GenBank登录号分别为: AM114925和AM114924), 并分析其基因组结构。运用生物信息学技术对测序结果进行分析显示: 基因组DNA由8个外显子和7个内含子组成, 与鸡基因序列同源率为60.8%~64.1%, 与人的同源率为42.9%。分子进化树进一步揭示了五龙鹅与鸡、其他鸟类、爬行类、哺乳类以及人类的进化关系, 同源建模分析发现该基因由氨基末端结构域和羧基末端结构域构成。     

Landscape-scale variation in the seed banks of floodplain wetlands with contrasting hydrology in China

1. At a local scale, the species composition, diversity and spatial variation of wetland plant communities are determined primarily by spatial and temporal heterogeneity in their environments. Less is known about variation at a landscape‐level. The floodplain of the Changjiang (Yangtze) River in China includes hydrologically connected, subtropical wetlands with different hydrological characteristics. 2. We examined seed‐bank species composition and richness in marshes of two contrasting hydrological types: permanent marshes, fed by local runoff, and lakeshore marshes more closely connected to the regulated river. Lakeshore marshes are flooded annually to depth of approximately 1 m and during flooding they support an alternate, aquatic vegetation type. The soil seed bank in March was a comparative estimator of species diversity. At the beginning of the growing season it included seeds from both phases of alternating vegetation types associated with the annual hydrological cycle. 3. A regional pool of 101 species was detected in the seed banks of six wetlands associated with the river and its tributaries: 56 occurred in permanent marshes and 59 in lakeshore marshes, with only 15 common to both. Species rarefaction curves indicated that more species occurred in permanent than lakeshore marshes at equal numbers of individuals sampled. However, the more heterogeneous lakeshore seed banks were estimated (Chao 2) to have greater total species richness (81) than permanent marsh (60). 4. Analysis using Sørensen's coefficient of similarity and DCA ordination revealed complex variation, with much greater differences between hydrological types than within them, irrespective of geographical distance. The types also differed significantly in the composition of four functional groups of species. 5. Despite the potential for dispersal of propagules via the annually pulsing river system (hydrochory), at a regional and landscape scale, diversity is maintained largely by large‐scale temporal hydrological heterogeneity and smaller scale spatial and topographic heterogeneity.     

间歇升温和热处理对‘紫李’果实采后生理的影响

以美国‘紫李’为试材,测定经间歇升温和热处理后果实的褐变度、抗氧化酶活性、抗氧化剂含量、膜质过氧化水平、总酚和可滴定酸含量的变化。结果表明:间歇升温和热处理可适当恢复因冷害而降低的细胞抗氧化活性,清除活性氧自由基,减少膜脂过氧化产物丙二醛的积累,抑制多酚氧化酶和过氧化物酶活性升高,强化抗低温防御系统,阻止多酚逆境代谢发生,使冷害和褐变症状得以延缓和减轻;同时,还可抑制可滴定酸含量的减少和固酸比的上升,延缓后熟衰老。间歇升温处理,李贮藏两个月果实品质良好。初步认为,-0.5～0℃贮藏,每15 d加温至18～20℃并保持l d,是贮藏美国‘紫李’适宜的变温模式。     

Separation and purification of Toxoplasma gondii tachyzoites from in vitro and in vivo culture systems

In this study, we evaluated four methods to separate and purify Toxoplasma gondii tachyzoites from in vivo and in vitro culture systems, including trypsin digestion, purification with a 3-μm filter, CF-11 cellulose purification, and Percoll purification. Our results indicate that both purification with a 3-μm filter and CF11 cellulose purification methods remove leukocytes or HeLa cells, and can therefore be used as candidate methods for the purification of in vivo and in vitro culture products. Trypsin digestion had a high tachyzoite recovery rate, but 22.35% of leukocytes and 69.64% of HeLa cells remained in the purified products. Percoll solution [30% (v/v)] also had a high tachyzoite recovery rate, but 3.44% of leukocytes and 61.61% of HeLa cells remained in the purified products. The 40% Percoll solution was also a candidate method for purifying tachyzoites from in vivo culture products, with a 65.45% tachyzoite recovery rate and without leukocytes.     

斜卧青霉L-06内切葡聚糖酶Ⅰ基因的克隆与表达

【目的】克隆斜卧青霉L-06的内切葡聚糖酶Ⅰ基因(egI),并实现其在大肠杆菌内的高效表达。【方法】利用RT-PCR技术克隆了斜卧青霉L-06的内切葡聚糖酶Ⅰ基因(egI),并将egI基因克隆到原核表达载体中,构建了重组质粒pET32a-egI。【结果】转化至大肠埃希菌Rosetta(DE3),经IPTG诱导重组蛋白表达,SDS-PAGE检测结果表明:重组表达产物的相对分子质量约为80 kD,与预期相符。重组表达的菌悬液,经破碎离心,取其上清液,进行纤维素酶活性染色,获得了活性条带。DNS法测得内切酶活力为2.56 IU/mL。【结论】构建了斜卧青霉L-06内切葡聚糖酶Ⅰ的原核表达系统。     

不同倍性香石竹杂交花粉管生长荧光显微观察及结实研究

以香石竹四倍体材料‘紫蝴蝶’(2n=4x=60)为母本,二倍体材料‘珍珠粉’和‘NH6’(2n=2x=30)为父本,利用荧光显微镜观察其授粉后花粉管生长情况,统计其座果率、亲和指数及种子萌发率,并对杂交后代进行倍性鉴定。结果表明,在‘紫蝴蝶’柱头上,‘珍珠粉’和‘NH6’的花粉2h开始萌发,花粉管多处出现胼胝质塞,且花柱组织出现胼胝质反应,4h花粉管到达柱头中部并出现胼胝质塞,6h花粉管到达柱头基部,17h柱头基部的花粉管增多,花粉管进入子房组织且子房组织出现胼胝质反应,17～24h花粉管能与胚珠结合,但结合率低;‘紫蝴蝶’ב珍珠粉’杂交未获得植株,‘紫蝴蝶’בNH6’杂交获得3株植株,染色体倍性鉴定表明3株植株均为四倍体,这可能是‘NH6’产生2n配子的缘故。     

西藏蚋属一新种（双翅目：蚋科）

记述西藏自治区蚋属Simulium蚋亚属Simulium 1新种,以陈汉彬教授的名命名为汉彬蚋Simulium(Simulium) hanbini sp. nov.。该新种与Simulium (Simulium) jingfui Cai & An, 2008及Simulium (Simulium)liubaense Liu & An, 2009相似,但该蚋爪无基齿,拉氏器较长,食窦的形状也不同。模式标本保存在北京军事医学科学院医学昆虫标本馆。