Effects of alismol isolated from Alismatis Rhizoma on calcium-induced contraction in the rabbit thoracic aorta

We examined the inhibitory effects of alismol, a sesquiterpenoid isolated from Alismatis Rhizoma, on vascular contractions induced by high concentrations of K+ and Ca2+, and on 45Ca2+ retention in normal and in high K+-containing medium. Alismol affected neither the resting tension nor the 45Ca2+ retention in normal medium, but it inhibited sustained contraction and increased 45Ca2+ retention induced by high K+ concentrations. Alismol did not affect norepinephrine-induced contractions in normal medium, nor phasic contractions in Ca2+-free medium. These results suggested that alismol inhibited mainly Ca2+ influx through a voltage-dependent Ca2+ channel.     

中稻田三种飞虱的捕食性天敌优势种及农药对天敌的影响

对两优0923非防治的中稻田白背飞虱、灰飞虱和褐飞虱与其天敌间的关系,采用灰色关联度法、生态位分析方法,对盆拍法调查的3种飞虱与其捕食性天敌在数量、时间和空间三方面关系进行分析,对每一种天敌对应的关联度、生态位重叠指数和相似性比例等参数标准化后的密切指数相加,按照密切指数值之和大小排序,评判3种飞虱捕食性天敌优势种.并用同样方法分析常规防治田农药对飞虱捕食性天敌的影响,以期为合理施药,科学保护和利用天敌优势种提供科学依据,其结果是,非防治田白背飞虱前三位天敌是条纹蝇虎、草间小黑蛛和锥腹肖蛸;灰飞虱的是八斑球腹蛛、茶色新园蛛和锥腹肖蛸;褐飞虱的是纵条蝇狮、四点亮腹蛛和黑肩绿盲蝽.防治田白背飞虱前三位天敌是条纹蝇虎、草间小黑蛛和锥腹肖蛸;灰飞虱的是拟水狼蛛、四点亮腹蛛和草间小黑蛛;褐飞虱的是黑肩绿盲蝽、拟水狼蛛和四点亮腹蛛.盆拍法的防治田和非防治田之间3种飞虱前三位的相同天敌,白背飞虱完全相同,灰飞虱没有相同天敌,褐飞虱的是黑肩绿盲蝽和四点亮腹蛛.飞虱与天敌在时间和数量关系上,扫网法的防治田和非防治田之间3种飞虱前三位的相同天敌,白背飞虱的是锥腹肖蛸和四点亮腹蛛,灰飞虱完全相同,褐飞虱的是纵条蝇狮和条纹影虎.两种稻田的差异主要是农药杀伤了飞虱,使飞虱数量减少,并对天敌有一定杀伤力,进而影响到天敌的发生规律.非防治田的盆拍法和扫网法之间,3种飞虱前三位相同的天敌,白背飞虱的是锥腹肖蛸,灰飞虱的也是锥腹肖蛸,褐飞虱的是纵条蝇狮.防治田两调查方法结果之间,白背飞虱的是锥腹肖蛸,灰飞虱和褐飞虱前3位天敌中没有相同天敌,其差异主要是两法调查稻株的部位不同所致.     

一株链霉菌菌株NCPC-1020中棘白霉素B脱酰基酶基因的克隆与功能分析

【目的】从一株土壤放线菌来源的野生型链霉菌菌株NCPC-1020中克隆一个具有棘白霉素B脱酰基酶活性的新基因。【方法】采用Degenerate和TAIL PCR两种方法,从链霉菌菌株NCPC-1020基因组中快速克隆获得了该基因序列,然后将基因在变铅青链霉菌TK24中进行异源表达,并进行全细胞催化底物脱酰基反应,采用LC-MS检测反应产物。【结果】LC-MS检测证实,棘白霉素B结构中脂肪链被酶促水解,从而证实该基因具有脱酰基酶活性。【结论】采用Degenerate以及TAIL PCR的方法能够快速获得未知功能的新基因。此基因的克隆,奠定了进行半合成棘白霉素类药物的研发基础。     

青藏高原矮嵩草草甸地下和地上生物量分配格局及其与气象因子的关系

基于2006—2015年青海海北站10年生物量及气候因子监测数据,分析了青藏高原高寒矮嵩草草甸生物量的季节及年际动态,并探讨了气候因子对其影响。结果表明:(1)季节尺度上,高寒矮嵩草草甸地上生物量表现为单峰变化曲线,8月为其峰值点,为(345.72±27.01) g/m~2,代表了高寒草甸的地上净初级生产力。而地下根系的现存量变化较为复杂,其中5—7月呈现持续上升趋势,8月快速下降,之后9月份急剧,且各月份之间未达到显著水平(P0.05);年际尺度上,10年间高寒矮嵩草草甸地上生物量整体呈现波动增加趋势,2014年为其峰值点,达(437.12±32.01) g/m~2。地下生物量呈现波动性变化,变异较大,10年间平均值为(2566.99±138.11) g/m~2;(2)高寒草甸光合产物分配主要分布在地下,80%地下根系生物量分布于地表0—10 cm土层,且不同土层根系生物量占总地下生物量的比值在不同月份较为稳定。(3)气候因子中,大气相对湿度是影响高寒草甸地上生物量大小的主要因素;而气候因子对地下根系生物量的影响极为微弱。研究表明,高寒嵩草草甸对环境变化具有较高的自我调节能力,且高寒草甸的演化受制于人类干扰,而非气候变化。     

湖北省马尾松精准适宜性空间分布

马尾松作为南方主要的造林树种,研究其适宜性精准空间分布对其退化评价、造林选地等具有生产指导价值。本研究采用多元线性回归克里金模型利用R语言对湖北省降雨量、气温等15个气象因子进行30×30m的空间插值,插值结果作为气象因子空间变量,插值的交叉检验结果≥90%。利用30m的DEM提取坡度、坡向、高程3个地形因子空间变量。首先利用年均温、最冷月最低温、年降水量、高程、土壤类型等限制性因子进行空间叠加排除不适宜的空间单元,然后在可能适宜的空间单元内,再基于18个空间变量和294个样方数据,利用最大熵模型(MAXENT)计算每个格网的适宜性指数,最后按照适宜指数进行适宜性分类。区别于以往大尺度区域研究,其结果是落实到地块尺度的精准计算。结果表明:ROC模型的评价效果达到较好水平(ROC=0.826),高程、土壤、降雨相对标准差、最湿季降雨量、坡度、年均温为湖北省马尾松的适宜性主要影响因子,马尾松中、强适宜区主要分布在湖北省东北部及中北部地区,主要分布在高程低于1200m、坡度在40°以下、年均温13-18℃的区域,中等及以上适生区面积共计4.65万km~2,占湖北省总面积的25.01%。     

Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting

Quantitative polymerase chain reaction-high-resolution melting (qPCR-HRM) analysis was used to screen for mutations related to drug resistance in Mycobacterium tuberculosis. We detected the C526T and C531T mutations in the rifampicin resistance-determining region (RRDR) of the rpoB gene with qPCR-HRM using plasmid-based controls. A segment of the RRDR region from M. tuberculosis H37Rv and from strains carrying C531T or C526T mutations in the rpoB were cloned into pGEM-T vector and these vectors were used as controls in the qPCR-HRM analysis of 54 M. tuberculosis strains. The results were confirmed by DNA sequencing and showed that recombinant plasmids can replace genomic DNA as controls in the qPCR-HRM assay. Plasmids can be handled outside of biosafety level 3 facilities, reducing the risk of contamination and the cost of the assay. Plasmids have a high stability, are normally maintained in Escherichia coli and can be extracted in large amounts.     

Interaction of two cis sites with the RNA replicase of the yeast L-A virus.

L-A is a 4.6-kilobase double-stranded RNA virus of Saccharomyces cerevisiae. The in vitro L-A replication reaction ((-)-strand synthesis) requires an internal site 400 bases from the 3' end in addition to the 3'-terminal 30 nucleotides of the L-A (+)-single-stranded RNA. Elimination of the internal site reduces the template activity 5-10-fold. Here we investigate how the internal site can stimulate the replication reaction which starts at the 3' end of the template. When these two sites are split into two distinct RNA molecules, the internal site can no longer stimulate replication (no trans-activation). However, establishment of an intermolecular hydrogen bonding between these RNAs restored the replication-enhancing activity of the internal site. This result is consistent with a model wherein L-A's RNA polymerase interacts first with the internal site and then with the 3' end site by either looping or by a local dissociation-reassociation mechanism. These results, however, clearly eliminate anchored tracking and sliding models which require continuity of the RNA molecule between these two cis sites.     

Isolation and characterization of a collagen binding domain in human von Willebrand factor

von Willebrand factor binds to fibrillar type I collagen in a rapid, temperature-independent, reversible, specific, and saturable manner. Evaluation of binding isotherms by Scatchard-type analysis demonstrated that 6-18 micrograms of von Willebrand factor bind per mg of collagen, with Ka between 2 and 8 X 10(8) M-1. Five distinct tryptic fragments, purified under denaturing and reducing conditions and representing over 75% of the molecular mass of the von Willebrand factor subunit, were tested for their capacity to inhibit the von Willebrand factor-collagen interaction. Complete inhibition was obtained with a 52/48-kDa fragment at a concentration of approximately 1 microM. The location of this fragment in the subunit was established to be between Val-449 and Lys-728. Fifteen monoclonal antibodies against the 52/48-kDa fragment inhibited von Willebrand factor binding to collagen. Six antibodies against other portions of the von Willebrand factor subunit had no inhibitory effect. The tryptic fragment was a competitive inhibitor of von Willebrand factor binding to collagen and, therefore, recognizes the same interaction site as the intact molecule. These studies precisely define a domain in the von Willebrand factor subunit that interacts with type I collagen.