Genetic Variation and Geographic Differentiation Among Populations of the Nonmigratory Agricultural Pest Oedaleus infernalis (Orthoptera: Acridoidea) in China

The nonmigratory grasshopper Oedaleus infernalis Saussure (Orthoptera : Acridoidea) is an agricultural pest to crops and forage grasses over a wide natural geographical distribution in China. The genetic diversity and genetic variation among 10 geographically separated populations of O. infernalis was assessed using polymerase chain reaction-based molecular markers, including the intersimple sequence repeat and mitochondrial cytochrome oxidase sequences. A high level of genetic diversity was detected among these populations from the intersimple sequence repeat (H: 0.2628, I: 0.4129, H_s: 0.2130) and cytochrome oxidase analyses (H_d: 0.653). There was no obvious geographical structure based on an unweighted pair group method analysis and median-joining network. The values of F_ST, θ^II, and Gst estimated in this study are low, and the gene flow is high (N_m> 4). Analysis of the molecular variance suggested that most of the genetic variation occurs within populations, whereas only a small variation takes place between populations. No significant correlation was found between the genetic distance and geographical distance. Overall, our results suggest that the geographical distance plays an unimpeded role in the gene flow among O. infernalis populations.     

Morphological and DNA Barcoding Evidence for Invasive Pest Thrips,Thrips parvispinus (Thripidae: Thysanoptera), Newly Recorded From India

South East Asia pest thrips species, Thrips parvispinus (Karny), is a serious pest on a number of agricultural and horticultural crops in a number of plant families. Based on an integrated approach of morphology and DNA barcoding, invasion of this serious pest is reported first time from India on papaya plantations. Molecular data have corroborated with the morphological identification. Haplotyping data suggested that the Indonesia may be a probable source of invasion of this pest to India.     

Species determination of Malaysian Bactrocera pests using PCR‐RFLP analyses (Diptera: Tephritidae)

BACKGROUND: Identification of Bactrocera carambolae Drew and Hancock, B. papayae Drew and Hancock, B. tau Walker, B. latifrons Hendel, B. cucurbitae Coquillett, B. umbrosa Fabricius and B. caudata Fabricius would pose a problem if only a body part or an immature stage were available. Analysis of polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) of cytochrome oxidase I (COI) gene using primers COIR, COIF, UEA7 and UEA10 and restriction enzymes (MseI, RsaI and Alu1) was carried out. The banding profiles in the electrophoresis gel were analysed. RESULTS: The COI gene in six Bactrocera spp. was successfully amplified by COIR and COIF, as well as UEA7 and UEA10, while B. caudata was amplified successfully only by UEA primers. Using COI amplified PCR products and restriction enzymes, distinct banding profiles for B. tau, B. latifrons, B. cucurbitae and B. umbrosa were observed, but not for B. carambolae and B. papayae. However, using UEA7, UEA10 and RsaI, B. caudata could be identified, while B. carambolae and B. papayae might possibly be separated from one another. It was also shown that adult body parts or immature life stages of B. carambolae, B. papayae, B. latifrons and B. cucurbitae produced the same banding profiles as the adults. CONCLUSION: PCR‐RFLP analyses are able to identify positively five Bactrocera species, while B. papayae and B. carambolae might possibly be separated from one another, even if immature life stages or adult body parts are used. Copyright © 2009 Society of Chemical Industry     

中国克虎天牛属整合分类

[目的]结合DNA条形码技术与形态学比较,研究并分类克虎天牛属Cleroclytus两近似种新疆克虎天牛Cleroclytus collaris Jakovlev,1885和沟胸克虎天牛Cleroclytus strigicollis Jakovlev,1900,为其准确鉴定提供研究依据.[方法]采用体视显微镜,观察比...     

2种大眼蟹线粒体COI基因序列的比较研究

测定了宽身大眼蟹(Macrophthalmus dilatatum)和日本大眼蟹(M.japonicus)线粒体细胞色素氧化酶I亚基(COI)基因片段的序列,其长度均为658bp,A、T、G、C的含量宽身大眼蟹为27.4%、30.9%、17.3%、24.4%,日本大眼蟹为25.8%、32.0%、17.4%、24.7%。宽身大眼蟹只有1种单倍型,日本大眼蟹出现2种单倍型,2种间有123(124)个变异位点,核苷酸差异率为18.69%(18.85%),其中转换68(69)处,颠换55(55)处,转换与颠换比约为1.2(1.3);种间差异远大于种内个体间差异(0.61%)。2种大眼蟹的AT(57.9%)含量明显高于GC含量,与其他蟹类COI基因研究结果相似。系统发生树的拓扑结构支持大眼蟹科与弓蟹科亲缘关系相对较近,二者为姊妹群关系。     

4种河蓝蛤线粒体COI和16S rRNA基因序列的种间遗传分析

对光滑河蓝蛤Potamocorbula laevis、黑龙江河蓝蛤P.amurensis、焦河蓝蛤P.ustulata、红肉河蓝蛤P.rubromuscula4个野生种共40个个体的线粒体COI和16SrRNA基因片段进行了扩增和测序,经过筛选和剪切,得到长度为650bp和450bp的片段。序列分析显示,序列的碱基组成中G+C含量较低,16SrRNA基因种间和种内的变异较低,COI基因片段种内和种间的变异较高。以沙海螂Mya arenaria为外群,用MEGA 4.0软件中的NJ法构建了系统进化树,通过遗传距离和系统进化树可以看出,4种河蓝蛤未能达到不同种之间显著的遗传分化。     

洪泽湖河蚬(Corbicula fluminea) 2种表型群体的遗传变异分析

采用形态学分析(壳长、壳宽和壳高)和分子标记技术(细胞色素氧化酶Ⅰ基因,COⅠ)对洪泽湖河蚬(Corbicula fluminea)黄色和黑色2个群体的形态和遗传多样性特征进行研究.形态参数统计分析结果显示,2个群体的形态特征之间有显著性差异.经PCR扩增和序列测定,获得614 bp COⅠ基因序列,2个群体的COⅠ基因序列的碱基组成高度一致,均表现出A+T的含量(64.8％)明显高于G+C的含量(35.2％).28个黑色个体发现7种单倍型,单倍型多样性和核苷酸多样性分别为0.794和0.04274;30个黄色个体发现5种单倍型,单倍型多样性和核苷酸多样性分别为0.607和0.02825.单倍型之间的遗传距离在0.002-0.091之间,其NJ和MP系统发生树表明,COⅠ基因单倍型聚为2个明显分支.分子方差分析(AMOVA)结果显示,Fst=0.21736 (P＜0.01),21.74％的变异来自群体间,78.26％的变异来自群体内,2个群体之间有显著的遗传分化.研究表明,应将洪泽湖河蚬黑色和黄色群体分别作为独立单元进行管理和保护.     

我国大陆近海裸胸鳝属鱼类新记录种 ——鞍头裸胸鳝(Gymnothorax sagmacephalus) 形态与分子系统学研究

为完善我国裸胸鳝属鱼类物种资源记录信息,本研究报道了采集我国大陆沿海水域(福建省厦门市、广东省汕头市)新记录种———鞍头裸胸鳝(Gymnothorax sagmacephalus B?hlke, 1997),目前该裸胸鳝世界上仅在日本有记录。通过对采集的样品进行详细的形态学特征分析,综合利用 DNA条形码COI基因及16S rRNA基因进行分子鉴定及系统分类研究。结果如下：鞍头裸胸鳝主要鉴别特征为:体色浅褐色,眼后头顶有一个暗色马鞍状的斑纹,在背鳍起始点前有一个大的黑色三角形斑块;腹部有黑色条纹从鳃孔下方延伸到肛门;背鳍、臀鳍边缘均为白色;体修长,全长为体高的19.06～19.60倍;牙齿均单行,上颌齿每侧8个,下颌齿12-14个;总脊椎数为162-168。基于COI及16S rRNA基因分析,鞍头裸胸鳝与其他形态接近、体色同为纯色的裸胸鳝遗传距离分别为：COI(0.115-0.226),16S rRNA(0.067-0.108),其中COI遗传距离已远大于Herbert设定的2%最小物种鉴定遗传距离,显示鞍头裸胸鳝为独立物种。进化树上,鞍头裸胸鳝与白边裸胸鳝聚为紧密姐妹分支,显示两者亲缘关系最为接近。研究结果有效阐明鞍头裸胸鳝在我国大陆沿海也有分布,为有效新记录种,为我国裸胸鳝属鱼类的系统分类及物种名录的修订提供相关分类基础。     

3种根萤叶甲实时荧光PCR检测技术研究

根萤叶甲属(Diabrotica)被我国列为进境植物检疫性有害生物,该属中的玉米根萤叶甲(D.virgifera virgifera)、十一星根萤叶甲(D.undecimpunctata)、巴氏根萤叶甲(D.barberi)是北美的主要农业害虫。为了建立一种快速的分子鉴定方法以鉴定这3种根萤叶甲,本研究从田间采集的成虫样品中获得其部分mtDNA COI序列,与Gen Bank中的相关序列进行比对,设计筛选出3种根萤叶甲的特异性引物和TaqMan探针,并进行实时荧光PCR特异性和灵敏度检测。特异性检测结果表明,用目标种根萤叶甲DNA和其特异性探针和引物进行实时荧光PCR反应时,在30个循环反应内(Ct值30)有近S型扩增曲线出现,同时其他种均无荧光信号增长。此外,灵敏度结果显示,玉米根萤叶甲和巴氏根萤叶甲可检测的最小DNA模板浓度,即实时荧光PCR反应的灵敏度为0.1 ng/μL,十一星根萤叶甲为0.01 ng/μL。     

DNA条形码技术在小花蝽属昆虫分类鉴定中的应用

本研究利用DNA条形码技术对13种小花蝽属Orius Wolff昆虫进行了鉴定,进行了59条COI基因序列碱基组成及种内、种间遗传距离的分析,采用邻接法、最大简约法、贝叶斯推论法构建了系统发育树。结果表明,小花蝽属昆虫COI基因序列碱基组成与典型的昆虫线粒体DNA一致,A+T平均含量(66.4%)明显高于G+C含量,密码子的第3位A+T含量高达90.7%,碱基替换多为同义替换;13种小花蝽种内平均遗传距离为0.008,种间平均遗传距离为0.128,种内、种间遗传距离没有重叠区域。3种方法构建的系统发育树的聚类分析与形态学鉴定结果基本一致,除微小花蝽Orius minutus(Linnaeus)可能存在隐存种现象外,其他同一种群的不同个体单独聚为一支。利用DNA条形码技术对小花蝽属昆虫进行物种快速分子鉴定具有可行性。