Sequence Analysis of mtDNA COIGene and Molecular Phylogeny of Different Geographical Populations of Bemisia tabaci (Gennadius)

Bemisia tabaci(Gennadius) is a serious pest in many cropping systems worldwide and occurs in different biotypes. The mtDNA COI gene of the 12 Bemisia tabaci (Gennadius) populations from different regions and countries were analyzed.Based on mtDNA COI sequences, their biotypes were characterized and phylogenetic relationships among these populations were established with the method of UPGMA. The results indicated the genetic similarity between those populations from Beijing, Zhengzhou, Zaozhuang, Nanjing, Shanghai, Haikou, and the B-biotype populations from California, Texas, Arizona reached 99.8-100%, which meant the nation-wide infested populations of B. tabaci in China in recent years were B-biotypes. Another population collected from Kunming of Yunnan Province showed very high similarity with Q-biotype B. tabaci from Spain and Morocco, which meant the Kunming population was Q-biotype. This is the first report on the invasion of Q-biotype into China.     

日照市农业标准化与产业化的调查与建议

当前农业的发展面临着需求与环境双重约束,农产品供给已由短缺转向总量基本平衡、丰年有余,市场需求对农业发展的约束作用越来越强;而另一方面,化肥、农药等化学合成物质的不合理施用,加重农产品的质量安全问题,影响农产品质量和效益的提高。进一步加强农业标准化工作,对改善农业生态环境,实施农业可持续发展具有十分重要的意义。     

石菖蒲根茎提取物β-细辛醚对谷蠹成虫的毒杀作用

采用药膜法、滤纸药膜选择法和三角瓶药纸熏蒸法分别测定了石菖蒲根茎提取物β-细辛醚(β-asa-rone)对谷蠹(Rhizopertha dominicaFab.)成虫的触杀、驱避及熏蒸作用。结果表明,β-细辛醚对谷蠹成虫具有较好的触杀与熏蒸活性。药膜法159.20μg/cm2浓度触杀处理72 h后对试虫的校正死亡率为100.00%。滤纸药膜选择法157.30、78.65、39.33、19.66μg/cm2剂量处理72 h内的平均驱避率均≥Ⅳ级。三角瓶药纸法166.67μL/L、83.33μL/L熏蒸处理72 h后试虫的校正死亡率分别为98.20%、75.86%。     

平原水网水体富营养化的限制因子研究

研究了光照、温度、营养(N,P)对平原水网水体藻类生长的影响.结果表明:在1 000～5 000 lx光强范围内,藻类的生长数量随光强的增加而呈对数关系增加,4 000 lx是最适宜的培养光照.在15～30 ℃区间内,藻类的生长数量也随着温度升高而增加,30 ℃是较适宜的温度.且温度的影响大于光照.水中T-P在0.1～0.75 mg/L范围内,随T-P浓度的提高,藻类数量呈对数关系增加,而水中T-N 浓度的变化对藻类生长的影响不是很明显.当水中T-P>0.5 mg/L、T-N>1.0 mg/L时,就能满足藻类大量增殖的需要,其中磷对藻类的影响大于氮.     

冷水江锑矿区重金属污染林地土壤环境质量评价

为了给锑矿区土壤生态修复、造林树种选择和快速恢复植被提供科学依据,采集了冷水江锑矿区受重金属污染的林地土壤样品,运用原子吸收分光光度法对土壤中6种重金属元素进行了测定,并对锑矿区受重金属污染的森林土壤环境质量进行了全面评价,结果表明:锑矿区三地点土壤中Sb、As、Pb、Hg、Cd、Zn含量有极显著差异。七里铺点土壤中以Sb、Hg、Cd、Zn含量最高,长龙界点土壤中以As含量最高,联盟点土壤中以Pb含量最高。随着土层厚度的增加,土壤中Sb、As、Pb、Hg、Cd、Zn含量呈递减的趋势。联盟点、长龙界点重金属污染的大小顺序是:Sb>Cd>Hg>As>Zn>Pb。七里铺点重金属污染的大小顺序是:Sb>Cd>Hg>Zn>As>Pb。依据内梅罗污染指数评价标准,冷水江锑矿区存在着严重的Sb、As、Cd、Hg和轻度的Zn、Pb复合污染。从不同土层重金属污染的内梅罗污染指数及重金属含量的分布特征分析,Sb、Hg在土壤中具有较大的迁移性,Zn具有中等的迁移性,而As、Pb、Cd则具有较小的迁移性。     

烟粉虱复合种不同地理种群的遗传分化

烟粉虱Bemisia tabaci(Gennadius) 是由具有遗传分化的不同地理种群组成的复合种。本文介绍了烟粉虱不同地理种群遗传分化的最新研究进展，并在世界各国对烟粉虱核糖体ITS1(rDNA ITS1)、线粒体COI（mtDNA COI）基因大量测序的基础上，进一步分析了烟粉虱不同地理种群的遗传分化。根据mtDNA COI和rDNA ITS1基因序列分析的结果，烟粉虱不同地理种群可分为5组，即亚洲组（Asia group）、美洲组（America group）、非洲组（Africa group）、澳洲组(Australia group)、B型/地中海/中东/北非/Ms型组(Biotype B/Mediterranean/Middle Eastern/Northern Africa/Biotype Ms group)；此外，还包括3个没有特定组的种群，即乌干达（Uganda）、科特迪瓦（Ivory Coast和台湾（Taiwan）种群。地理隔离可能是造成烟粉虱不同地理种群遗传分化的最重要驱动力。许多具有入侵性或生物学优势的烟粉虱种群随着人类活动而传入新的地区，造成了严重的经济损失。有必要加强烟粉虱生物型的监测，遏制已入侵烟粉虱种群的蔓延，防止新的具有潜在危险性的烟粉虱种群传入中国。     

猕猴桃GalUR表达与抗坏血酸积累的关系

 通过RT-PCR从美味猕猴桃（Actinidia deliciosa）果实中克隆了D–半乳糖醛酸还原酶（GalUR）全长 cDNA序列,并分析其序列特征,进行原核表达,制备特异抗体,分析其mRNA及蛋白表达水平与果实发育过程及不同组织中抗坏血酸（AsA）合成和积累的关系。结果表明：克隆的GalUR cDNA（GenBank登录号为GU339035）包含的最大开放阅读框（open reading frame,ORF）为990 bp,编码329个氨基酸残基,预测分子量为37 kD,与报道的草莓等其他植物GalUR具有较高的相似性。构建的pET-32a（＋）-GalUR载体在大肠杆菌BL21（E. coli BL21）中异源表达后,获得了主要以包涵体存在约58 kD的融合蛋白GalUR-His（His约21 kD）。以该蛋白制备抗体,与重组蛋白的Western杂交表明该抗体能与GalUR蛋白发生特异反应。对猕猴桃可溶性蛋白杂交显示,猕猴桃体内GalUR蛋白的分子量约37 kD。在猕猴桃不同组织和发育过程中,GalUR表达水平与AsA含量表现为高度一致,但前体饲喂发现,与L–半乳糖相比,D–半乳糖醛酸并不能有效促进猕猴桃AsA的合成。这些结果表明以单体形式存在的GalUR与猕猴桃AsA合成有着密切关系,但它可能并不是猕猴桃AsA积累的主要调控环节。     

Effect of ROCK1 and ROCK2 silencing by shRNA on hypoxia-induced apoptosis of rat cardiomyocytes

AIM: To investigate the effects of Rho-associated coiled-coil protein kinase-1 (ROCK1) and ROCK2 on apoptosis induced by hypoxia in rat cardiomyocytes. METHODS: Rat cardiomyocytes were cultured primarily and identified using an antibody targeting α-actin of striated muscle. ROCK1-shRNA and ROCK2-shRNA were transiently transfected into the cells by liposome. After 48 h, these cells were subject to hypoxia for 6 h. The cells were divided into 5 groups: blank control group, hypoxia group, hypoxia+negative control shRNA group, hypoxia+ROCK1-shRNA group and hypoxia+ROCK2-shRNA group. The beating frequency and rhythm of the cardiomyocytes were assessed by microscopy. The activity of lactate dehydrogenase (LDH) in the cell culture supernatants was detected by automatic biochemical analyzer. The cell survival rate was analyzed by the method of MTT. The cell apoptotic rate was assessed by flow cytometry. Western blotting was used to determine the expression of ROCK1, ROCK2, caspase-3 and p-PI3K. RESULTS: The primary culture of the cardiomyocytes was successful. Western blotting results showed that the transfection of ROCK1-shRNA or ROCK2-shRNA decreased the expression of ROCK1 or ROCK2 in the cardiomyocytes. Hypoxia slowed down the beat frequency of the cardiomyocytes, also made the rhythm disorder. Hypoxia increased the release of LDH and decreased the cell survival rate. Flow cytometry results showed that hypoxia increased the cell apoptotic rate. Hypoxia increased the expression of caspase-3 and decreased the expression of p-PI3K. Transfection of ROCK1-shRNA and ROCK2-shRNA into the cardiomyocytes reduced all the effects of hypoxia mentioned above. CONCLUSION: Down-regulation of ROCK1 and ROCK2 expression suppresses the apoptosis of rat cardiomyocytes induced by hypoxia. The mechanism is associated with the inhibition of caspase-3 activation and the up-regulation of p-PI3K expression.     

Construction of a lentiviral RNA interference vector targeting rat myocardin and its effect on vascular smooth muscle cell differentiation

AIM: To construct a lentiviral RNA interference(RNAi)vector targeting rat myocardin mRNA and to investigate its effect on the differentiation of vascular smooth muscle cells(VSMCs).METHODS: Three pairs of dsDNA targeting rat myocardin mRNA were designed, synthesized and cloned into lentiviral vector pGCSIL-GFP to generate pGCSIL-GFP-shMyocd lentvirus. A Flag-tagged myocardin-overexpression vector pEGFP-N1-Myocd was constructed with pEGFP-N1/X124G. After these two vectors were cotransfected into 293T cells, the flag protein was assessed by Western blotting to analyze the knockdown effect of pGCSIL-GFP-shMyocd. The expression of myocardin and SM22α was also detected by RT-PCR and Western blotting after the pGCSIL-GFP-shMyocd viruses were transfected into primary cultured rat aortal VSMCs.RESULTS: The rat myocardin lentviral RNAi vector pGCSIL-GFP-shMyocd and myocardin-overexpression vector pEGFP-N1-Myocd were successfully constructed. After these two kinds of vectors were cotransfected into 293T cells,the No.1 interfering vector displayed the highest inhibitory effect on flag expression.After the No.1 lentvirus at the titer of 1×10¹² TU/L was transfected into VSMCs, the myocardin and SM22α expression was significantly attenuated. CONCLUSION: The lentiviral pGCSIL-GFP-shMyocd RNAi vector is successfully constructed, which is useful for further study regarding the molecular mechanism of the phenotypic switching in VSMCs under special pathological conditions such as atherosclerosis. Inhibition of myocardin expression in VSMCs leads to the decrease in the expression of differentiation marker, and implies a crucial role of myocardin in VSMCs differentiation.     

温敏型核雄性不育系番茄T4育性的研究

以温敏型核雄性不育系番茄T4及对照First为试材,通过调查花粉发芽率、自交结实率、果实种子数以及采用Tricine-SDS-PAGE电泳方法分析叶片及花药蛋白质的变化,研究T4的育性转化,并分析其在可育和不育条件下花粉发芽率、自交结实率、果实种子数、叶片及花药蛋白质的差异。结果表明：28 ℃/18 ℃（昼/夜）处理的T4花粉发芽率、自交结实率最高,果实种子数最多,与对照First差异不显著,表明此时T4恢复部分育性|28 ℃/24 ℃和28 ℃/12 ℃处理的T4花粉发芽率、自交结实率和果实种子数与对照差异均达到极显著水平,表现为不育。不同温度处理的T4花药蛋白质与叶片蛋白质差异均达到显著水平|28 ℃/24 ℃和28 ℃/12 ℃处理的T4叶片蛋白质与对照差异显著,但28 ℃/18 ℃处理的差异不明显。