高效氯氰菊酯不同汰选频度条件下桔小实蝇高抗品系抗药性发展动态

为了研究高效氯氰菊酯不同施用强度对桔小实蝇Bactrocera dorsalis(Hendel)高抗品系抗药性发展动态的影响,为指导田间科学合理使用高效氯氰菊酯防治该虫提供理论依据,本文以室内培育的桔小实蝇对高效氯氰菊酯高抗品系成虫为研究对象,以高效氯氰菊酯对敏感品系的毒力作为毒力基准线,按高效氯氰菊酯汰选间隔时间长短设置30 d、60 d、90 d、120 d 4个处理,以在不接触药剂常规条件下饲养的高抗品系作对照,采用药膜法进行抗药性汰选和毒力测定,每30天试验1次,共计试验10次,获得致死中浓度(LC_(50))和抗性倍数(R_m),分析抗性发展动态与汰选间隔时间的关系。研究结果表明270 d后,不同间隔时间长度汰选处理,桔小实蝇高抗品系对高效氯氰菊酯的抗药性发展动态存在明显差异,总体表现为汰选间隔时间越短,抗性增长越快。30 d汰选1次处理,高效氯氰菊酯对桔小实蝇成虫的致死中浓度从第1次的582.7 mg/L上升到1133.6 mg/L,抗性倍数从98.0倍上升到190.7倍;60 d汰选1次处理,致死中浓度上升为828.0 mg/L,抗性倍数上升为139.3倍;90 d汰选1次处理,致死中浓度为529.2 mg/L,抗性倍数为89.0倍;120 d汰选1次处理,致死中浓度、抗性倍数分别为511.3 mg/L、86.0倍;未进行汰选的抗性品系致死中浓度由582.7 mg/L下降到368.1 mg/L,抗性倍数也从98.0倍下降到61.9倍。建立了270 d后高效氯氰菊酯汰选间隔时间与桔小实蝇成虫抗药性增长比率之间的关系方程为Y=11.427X~(-0.529)。由实验室模拟实验结果预测当果园中连续两次使用间隔时间在99 d以上时,应可降低桔小实蝇对高效氯氰菊酯的抗性继续上升的风险。     

桔小实蝇抗多杀霉素的生物适合度代价

桔小实蝇Bactrocera dorsalis ( Hendel) 是中国南方地区果蔬重要害虫之一,严重威胁农业生产。多杀霉素是防治桔小实蝇的重要药剂之一,近年来由于在农业生产中大量使用,田间防治效果已有所下降,已出现了抗药性现象。为明确桔小实蝇对多杀霉素的抗药性发展规律,本研究比较了桔小实蝇敏感品系和抗性品系的发育、生殖和存活差异。结果显示：抗多杀霉素桔小实蝇品系具有死亡率增加、部分虫态发育历期延长和繁殖力下降等适合度代价。抗性品系的内禀增长率由0.19下降至0.15,相对适合度降至0.62,表明桔小实蝇对多杀霉素存在较低的抗性风险。     

不同地理区域桔小实蝇解毒酶系活性及其与抗药性水平关系

昆虫体内的解毒酶活性是反映其抗药性水平的主要生理指标，本文比较了桔小实蝇Bactrocera dorsalis的几种解毒酶活性和抗药性水平关系。测定了9个地理品系和相对敏感品系的桔小实蝇成虫的三种解毒酶，即多功能氧化酶(Mixed function oxidase, MFO)、羧酸酯酶（Carboxylesterase, CarE）和谷胱甘肽Ｓ-转移酶(Glutathione S-transferase, GST)的活性,利用药膜法测定其对敌百虫(Trichlorphon)、高效氯氰菊酯(β-cypermethrin)和阿维菌素(Avermectin)的抗性水平，比较了各地理品系的桔小实蝇这些酶活性和对不同杀虫剂的抗性水平的关系，并作通径分析。结果表明：广东广州地区桔小实蝇体内的MFO-O-脱甲基活性最高，为相对敏感品系酶活性的1.4782倍；而广东茂名地区品系酶活性最低，只有0.8649倍。广东惠州地区桔小实蝇体内CarE活性最高，为相对敏感品系酶活性1.8147倍；而广西南宁地区的桔小实蝇体内酶活性最低，为敏感品系的0.9636倍。广东茂名地区桔小实蝇GST活性最高，是相对敏感品系2.2557倍；而广东广州地区桔小实蝇GST活性最低，只有相对敏感品系的1.1622倍。抗性水平表明：各地理品系对敌百虫的抗性水平为相对敏感品系的2.1552倍至100.2271倍之间，对高效氯氰菊酯的抗性水平在1.0065到26.0026倍之间，对阿维菌素的抗性水平在2.3353倍至29.0688倍之间。相关性分析表明：桔小实蝇体内的GST活性和对敌百虫抗性水平的相关系数为0.41，存在显著性正相关；CarE活性与桔小实蝇对高效氯氰菊酯抗性水平存在极显著正相关，相关系数为0.50。通径分析结果表明：GST活性对敌百虫的抗药性水平的直接通径系数为0.4414，对敌百虫的抗性上升起到正向作用；MFO-O-脱甲基活性、CarE活性和GST活性对高效氯氰菊酯抗性水平的直接通径系数分别为0.3311，0.4946和0.1775，均起到正向作用；GST活性与阿维菌素的抗性水平的直接通径系数很小，为0.0668。结果显示了桔小实蝇的解毒酶与抗药性水平关系密切，在抗性发展中起到了促进作用。     

茚虫威对不同抗药性品系小菜蛾呼吸代谢的影响

以小菜蛾Plutella xylostella(L.)敏感品系(S)、田间品系(F)及茚虫威汰选抗性品系(T17)幼虫为供试虫源,采用便携式光合作用测定仪测定小菜蛾3～4龄幼虫在受药前后或不同受药剂量等条件下呼吸速率的变化,从能量代谢的角度研究抗药性机制。结果表明,汰选抗性品系、田间品系与敏感品系同龄期小菜蛾幼虫在未受药条件下的呼吸速率差异不显著,表明抗药性的产生并未影响小菜蛾本底的呼吸速率。以各个品系的茚虫威LC20和LC50剂量处理幼虫后,3个品系的呼吸速率均明显提高,汰选抗性品系呼吸速率提高幅度明显大于其他2个品系。在LC20剂量下3个品系呼吸速率峰值均出现在2h前后,10h后恢复到正常水平;在LC50剂量下敏感品系没有明显差异;而田间品系和汰选抗性品系分别在药剂处理后4h和6h达到呼吸高峰,汰选抗性品系保持高水平呼吸速率时间长达9h,分别于药后15h和24h恢复到正常水平。这表明用药后抗性品系呼吸速率的提高幅度与小菜蛾的解毒代谢能力有关,这也揭示了昆虫幼虫中毒后能量消耗会随着抗性水平的提高而增加。     

桔小实蝇对多杀霉素的抗药性风险评估

桔小实蝇Bactrocera doralis (Hendel)是华南地区果蔬重要害虫,近年来危害严重,对农业造成巨大的经济损失.本研究采用阈性状分析方法,对桔小实蝇敏感品系对多杀霉素抗性进行选育,计算了桔小实蝇对多杀霉素的抗性现实遗传力,评估了抗药性产生的风险.结果表明:经过16代选育,桔小实蝇对多杀霉素抗性增加了7.05倍(LC50从0.60 mg/L增加到4.23 mg/L),现实遗传力为0.1774.结果显示桔小实蝇对多杀霉素抗性发展较慢.     

桔小实蝇实验种群对敌百虫、高效氯氰菊酯和阿维菌素的抗性增长规律

桔小实蝇Bactrocera doralis (Hendel)是华南地区水果蔬菜上重要害虫,近年来暴发成灾,造成巨大的经济损失。使用化学杀虫剂是防治该虫的重要方法。在长期、频繁接触杀虫剂条件下该虫的抗药性产生、发展规律等问题值得深入研究。本文采用LC₅₀、LC_80-90两种剂量汰选成虫、幼虫,研究了桔小实蝇对敌百虫、高效氯氰菊酯和阿维菌素3种药剂的抗性增长规律。通过不接触药剂继代饲养获得了对3种杀虫剂的敏感品系,建立了毒力回归方程,明确了毒力敏感基线LC₅₀分别为1.6024 mg/L,3.0964 mg/L和0.6074 mg/L。不同药剂种类、不同汰选方式时桔小实蝇的抗药性发展速度是不同的。以成虫死亡率50%左右作为选择压力,桔小实蝇对敌百虫的抗性发展最快,汰选至14代抗性增长至84.6倍;其次是高效氯氰菊酯,抗性增长至27.1倍;而对阿维菌素的抗性仅上升至7.7倍 。在成虫和幼虫80%～90%死亡率左右的选择压力下,桔小实蝇对高效氯氰菊酯的抗性发展最快,汰选14代后抗性增长至125.4倍; 其次是敌百虫,抗性增大至49.9倍;对阿维菌素抗性增长相对较缓慢,11代后为17.9倍。在成虫80%～90%死亡率左右的选择压力下,汰选14代、14代、11代后桔小实蝇对高效氯氰菊酯、敌百虫、阿维菌素分别增大至58.7倍、37.6倍、11.9倍。对成虫、幼虫均进行汰选桔小实蝇抗药性发展更快。建立了不同汰选方式时桔小实蝇实验种群汰选代数与对3种杀虫剂的抗性程度之间的关系方程,其中高效氯氰菊酯、敌百虫处理符合韦布尔曲线模型,阿维菌素处理符合幂曲线方程。     

基于性信息诱捕监测广东桔小实蝇和瓜实蝇发生动态及其对2种常用农药的抗药性评价

利用性信息素诱捕的方法对广东省茂名、佛山、梅州和韶关的桔小实蝇Bactrocera dorsalis (Hendel)和瓜实蝇Bactrocera cucurbitae (Coquillett)种群动态进行监测;并用药膜法测定了两种实蝇不同地理种群成虫对阿维菌素和甲维盐的抗药性情况。结果表明,桔小实蝇和瓜实蝇在所监测的4个地市全年均有发生,桔小实蝇和瓜实蝇种群数量发生高峰期集中在5月中旬至8月中旬,从4月中旬开始田间种群数量开始增加,10月中旬以后种群数量急剧下降,高峰期与果实成熟期基本吻合。抗药性监测结果表明,桔小实蝇和瓜实蝇对阿维菌素和甲维盐的抗药性有逐年上升趋势,广东4个地市桔小实蝇和瓜实蝇已对阿维菌素产生中等抗性水平;2019年11月监测梅州地区桔小实蝇种群和韶关瓜实蝇种群对甲维盐抗性倍数分别为4.32和3.42倍,仍处于敏感水平,其余地区种群对甲维盐均达到了低水平或中等水平抗性。     

蝇类抗性品系培育的研究进展

随着杀虫剂的广泛应用,农业、卫生类病媒生物抗药性日益严重,不仅影响果蔬生产,危害人类健康,更带来环境污染和生态破坏等问题.昆虫抗性品系培育作为研究抗药性机制和抗性治理的一种重要实验室研究手段,越来越受到研究者的关注.蝇是一类分布广泛的公共卫生和农业害虫,通过抗性品系培育研究也可为有害蝇类的抗性机制研究和虫害防制提供可靠...     

桔小实蝇抗敌百虫品系的实验种群生物学比较研究

【目的】研究明确桔小实蝇 Bactrocare dorsalis (Hendel) 抗药性品系与敏感品系的基本生物学特征以及相对适合度、内禀增长率等种群生物学参数差异。【方法】实验室条件下筛选出对敌百虫具中抗和高抗的桔小实蝇品系,观察记录其与敏感品系的生物学特性,比较三者之间的主要生物学参数。【结果】桔小实蝇敏感品系与2个抗性品系的卵、幼虫和蛹的发育历期、化蛹率、产卵前期、雌雄虫寿命、存活时间及雌虫比率均无显著差异。中抗品系卵孵化率和成虫羽化率分别为68.33%和93.73%,均显著低于敏感品系（分别为88.33%和97.93%）和高抗品系（分别为86.67%和98.21%）。与敏感品系的产卵量864.61粒/雌和高抗品系的产卵量750.70粒/雌相比,中抗品系的产卵量降低至630.87粒/雌。3个品系雌虫日产卵量动态规律比较一致,均呈开始产卵后短时间内进入产卵盛期,继而产量达到高峰,之后产卵量波动下降。但是与敏感品系相比,2个抗性品系雌虫较早进入产卵高峰。对成虫存活率动态拟合方程参数c值均大于1,符合I型存活曲线基本模型,表明在实验室理想条件下3个品系成虫均能达到其平均寿命。敏感品系种群趋势指数（I）最高,为339.41,其次为高抗品系（I =307.82）,最小为中抗品系（ I =175.79）,表明对敌百虫产生抗性抑制了桔小实蝇种群的增长,中抗品系抑制程度更大。敏感和高抗品系实验种群净增殖率较高,分别为327.89和284.29;中抗品系显著较低,为217.49。2个抗性品系内禀增长率（r_m）和周限增长率（λ）间无显著差异,且均显著大于敏感品系,世代历期（T）则显著缩短。基于种群净增殖率获得的敌百虫高抗和中抗品系相对适合度分别为0.8670和0.6633。【结论】在敌百虫的选择压力下,桔小实蝇抗药性品系的世代周期、中抗品系卵的孵化率和蛹的羽化率显著降低,其他多项生物学特征无显著变化。抗性品系的繁殖力和种群世代增长量受到抑制,以中抗品系更为明显;但与敏感品系相比,抗性品系种群增长潜力更大。     

桔小实蝇抗性个体流动对抗性个体频率的影响

为了解害虫个体的迁移对害虫群体抗药性发展的影响, 本研究利用实验室内建立的桔小实蝇Bactrocera dorsalis对敌百虫和高效氯氰菊酯两种抗性品系以及相对敏感品系, 设计5%~25%个体的迁移比例, 研究桔小实蝇抗、 感个体流动对原始种群抗性个体频率的影响。结果表明: 桔小实蝇抗性个体迁入敏感种群, 使得敏感种群抗性个体频率增加, 在抗性个体迁移率为25%时, 影响敏感种群敌百虫和高效氯氰菊酯抗性个体频率的变化值分别为20.04%和41.75%; 同样随着敏感个体迁入比例的增加, 抗性种群中抗性个体频率降低程度越大, 在敏感个体迁移率为25%时, 抗性种群敌百虫和高效氯氰菊酯抗性个体频率的变化值分别为56.20%和25.88%。利用抗性个体频率变化值与相应迁移率的比值来表示迁移的相对效率, 在抗性个体迁移率为5%时, 影响敏感种群抗性个体频率变化的相对效率最高; 在敏感个体迁移率分别为5%和10%时, 影响敌百虫抗性种群和高效氯氰菊酯抗性种群抗性个体频率变化的相对效率最高。以抗、 感个体迁移引起的抗性个体频率变化值进行趋势拟合, 发现抗敌百虫与抗高效氯氰菊酯桔小实蝇品系不同迁移比例下的抗性个体频率变化趋势分别符合密度模型(density model)和房屋模型(housing model), 相关系数分别是0.9696和0.9647。研究结果表明通过合理地设计抗、 感迁移比例能有效地延缓桔小实蝇抗性水平的上升, 达到抗性治理的目的。     

微卫星(TATG)n基序在香菇菌种中的验证

以（TATG）4重复序列为引物对香菇属的3个种13个菌株的微卫星区DNA进行PCR扩增,15%的琼脂糖凝胶电泳,获得了25个条带,并且在供试菌株上表现出多态性,可以实现遗传分类研究。为了验证微卫星分子标记实验准确性,又用RAPD技术对13个供试菌株进行了实验。7个引物在13个菌株上共获得了102条多态性条带。通过聚类分析,RAPD获得的分类结果与微卫星分子标记获得的结果一致。此外,为了证明微卫星分子标记获得的条带不是假阳性,在实验中回收了No.10菌株的PCR扩增产物,进行克隆测序。测序结果显示有（TATG）n基序存在,并且达到了微卫星基序重复数量的最低限度。通过本实验可知,香菇中是存在微卫星（TATG）n基序的, 且基序的多态性可以用于香菇的遗传分类研究。     

应用微卫星标记对雌核发育银鲫的遗传多样性初探

利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼（Cyprinus carpino.L)的8个微卫星DNA标记,对银鲫（Carassius auratus gibelio Bloch)的5个不同雌核发育系的24尾个体进行PCR扩增。分析电泳结果发现,除MFW28未能在银鲫中稳定地扩增出相应的同源序列,其余的7对引物扩增的重复性和稳定性都很好,随引物不同,各等位基因数为1-14个,大小在100-506bp。在MFW1、MFW4、MFW19、MFW20、MFW23和MFW246个微卫星的扩增图谱中,不同的雌核发育系扩增出各自独特的图谱,而同一系内的不同个体间具有高度的遗传同质性,但仍然在个别个体中检测到少量的多态片段。不同系间的扩增图谱呈现出高度的遗传异质性,共鉴定出23个可以用于有效区分5个不同雌核发育系的分子标记。这5个微卫星标记反映了银鲫5个雌核发育系间的相互亲缘关系,其中P和A系同属一个雌核发育系,F系起源于E系,A、D和E系可能分别独立地起源于不同的杂交事件,鉴定的微卫星分子标记为进行银鲫群体遗传学和进化遗传学研究,以及银鲫的分子标记育种和进行基因组作图提供了理想的工具。     

Living in a jar: genetic variation and differentiation among laboratory strains of the red flour beetle

The red flour beetle, Tribolium castaneum, is a common pest, which has become an important model study organism, especially in genetic, ecological and evolutionary research. Although almost all studies on this species have been conducted using established laboratory strains, very little is known about the loss of genetic diversity within the strains and genetic divergence between different laboratory stocks. In this study, five long‐term laboratory strains and one wild strain were examined for genetic variation at 12 microsatellite loci, which were designed using publicly available sequences. One of the laboratory strains is resistant to phosphine and one to organophosphorous insecticides. All strains had significant amounts of molecular variation, but genetic diversity in the laboratory strains was lower than in the wild‐derived strain used as control. We observed significant molecular divergence among the strains, however, the relationship between them reflected resistance status rather than geographic origins. We found no evidence for recent bottlenecks, but the wild‐derived population showed signs of demographic expansion. A novel multivariate method, multiple co‐inertia analysis, revealed that the two loci contributing most to the divergence between the resistant strains were located on the eighth chromosome, near genes associated with insecticide resistance.     

Molecular phylogeny and population structure of the codling moth (Cydia pomonella) in Central Europe: I. Ancient clade splitting revealed by mitochondrial haplotype markers

The codling moth (Cydia pomonella L., Tortricidae, Lepidoptera) is an important pest of pome fruit with global distribution. It has adapted successfully to different habitats by forming various ecotypes and populations, often termed strains, which differ among each other in several morphological, developmental, and physiological features. Many strains of Cydia pomonella have developed resistance against a broad range of chemically different pesticides. Obviously, pesticide-resistant strains must have a genetic basis inherent to the gene pool of codling moth populations, and this deserves our particular attention. The primary intention of the present study was to contribute novel information regarding the evolutionary phylogeny and phylogeography of codling moth populations in Central Europe. In addition, we aimed at testing the hypothesis that differential biological traits and response patterns towards pesticides in codling moth populations may be reflected at a mitochondrial DNA level. In particular, we wanted to test if pesticide resistance in codling moths is associated repeatedly and independently with more than one mitochondrial haplotype. To this end, we analyzed mitochondrial DNA and constructed phylogenetic trees based on three mitochondrial genes: cytochrome oxidase I (COI), the A+T-rich region of the control region (CR), and the nicotinamide adenine dinucleotide dehydrogenase subunit 5 (ND5). The results indicate that Central European populations of Cydia pomonella are clearly divided in two ancient clades. As shown by means of a molecular clock approach, the splitting of the two clades can be dated to a time period between the lower and middle Pleistocene, about 1.29-0.20 million years ago. It is assumed that the cyclic changes of warm and cold periods during Pleistocene may have lead to the geographic separation of codling moth populations due to glaciation, giving rise to the formation of the two separate refugial clades, as already shown for many other European animal species. Due to their inclination towards developing novel detoxification gene variants, codling moth individuals from both clades independently and multifariously may have developed pesticide resistance, and this process may be ongoing. During their more recent evolutionary history, natural events such as the gradual disappearance of climate-specific geographic barriers, as well as human-aided dispersal in recent historic times, may have allowed codling moth haplotypes from the original clades to interbreed and completely merge again, creating a globally successful insect species with a gene pool capable of responding to novel selective challenges by rapid adaptation.     

Detected microsatellite polymorphisms in genetically altered inbred mouse strains

Microsatellites are 50–200 repetitive DNA sequences composed of 1- to 6-base-pair-long reiterative motifs within the genome. They are vulnerable to DNA modifications, such as recombination and/or integration, and are recognized as “sentinel” DNA. Our previous report indicated that the genotypes of the microsatellite loci could change from mono- to poly-morphisms (CMP) in gene knockout (KO) mice, implying that genetic modification induces microsatellite mutation. However, it is still unclear whether the random insertion of DNA fragments into mice genomes produced via transgene (Tg) or N-ethyl-N-nitrosourea (ENU) would also result in microsatellite mutations or microsatellite loci genotypes changes. This study was designed to find possible clues to answer this question. In brief, 198 microsatellite loci that were distributed among almost all of the chromosomes (except for the Y) were examined through polymerase chain reaction to screen possible CMPs in six Tg strains. First, for each strain, the microsatellite sequences of all loci were compared between Tg and the corresponding background strain to exclude genetic interference. Simultaneously, to exclude spontaneous mutation-related CMPs that might exist in the examined six strains, mice from five spontaneously mutated inbred strains were used as the negative controls. Additionally, the sequences of all loci in these spontaneous mutated mice were compared to corresponding genetic background controls. The results showed that 40 of the 198 (20.2 %) loci were identified as having CMPs in the examined Tg mice strains. The CMP genotypes were either homozygous or heterozygous compared to the background controls. Next, we applied the 40 CMP positive loci in ENU-mutated mice and their corresponding background controls. After that, a general comparison of CMPs that exist among Tg, ENU-treated and KO mouse strains was performed. The results indicated that four (D11mit258, D13mit3, D14mit102 and DXmit172) of the 40 (10 %) CMP loci were shared by Tg and KO mice, two (D15mit5 and D14mit102) (5 %) by Tg and ENU-treated mice, and one (D14mit102) (2.5 %) by all three genetic modifications. Collectively, our study implies that genetic modifications by KO, Tg or chemical mutant can trigger microsatellite CMPs in inbred mouse strains. These shared microsatellite loci could be regarded as “hot spots” of microsatellite mutation for genetic monitoring in genetic modified mice.     

Polymorphic microsatellite loci of the rat (Rattus norvegicus)

The EMBL and GenBank DNA databases were searched for microsatellite sequences of the rat containing dinucleotide repeats of (CA)_n and (GA)_n. Among those obtained, 23 sequences were analyzed by polymerase chain reaction to examine the size variation of the amplified fragment in inbred rat strains. All of the 23 microsatellite sequences varied in size among the strains tested. The 23 microsatellite loci in a pair of substrains separated from the same progenitor strain were then analyzed. Fragments identical in size were observed in all loci of the two substrains, indicating the stability of the microsatellite over a large number of generations. The microsatellite loci, therefore, should be useful markers for linkage analyses in the rat.     

Discrimination of reproductive forms of Thrips tabaci (Thysanoptera: Thripidae) by PCR with sequence specific primers

In agriculture, although it is important to identify species of pest insects, the morphological identification is often difficult. DNA genotyping is useful for the identification of species in morphologically indiscriminable species. Thrips tabaci (Lindeman) can be divided into two reproductive forms (arrhenotoky and thelytoky, each of which different in pesticide resistance) but morphological discrimination is not possible. Here, we establish a simple method to discriminate the strains based on their mitochondrial DNA sequences. Phylogenetic analysis including the T. tabaci and congeneric species provided ancestor sequences of each strain of T tabaci. Based on the ancestor sequences, we developed a primer set that include strain specific primers on sense strand and common primer on anti sense strand. Using this primer set, the strains of 196 individuals of T. tabaci were successfully assigned to each ofgenotypic forms. As the phylogeny and ancestor sequences were based on worldwide samples, this method will work well on most populations around the world.     

Detection of genetic polymorphism among and within Echinococcus granulosus strains by heteroduplex analysis of a microsatellite from the U1 snRNA genes

Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNA gene complex generated a multiple band pattern due to the priming of paralogous sequences. Denaturation and slow renaturation of polymerase chain reaction products allow the formation of heteroduplex DNA that can be detected by its differential mobility in polyacrylamide gel electrophoresis. Heteroduplex analysis was used to determine if the U1 snRNA microsatellite could be a useful genetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragment from E. granulosus was isolated and characterized by Southern blot and sequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep, cattle, and camel strains. The former two showed polymorphism and shared three of the six patterns found for sheep strain. The cattle strain displayed two patterns, and the camel strain was monomorphic. The electrophoretic profiles were used for statistical analysis in order to determine genetic distance and the relationship among strains. Heteroduplex analysis can be helpful in genotyping E. granulosus strains and is useful in detecting polymorphism within strains.     

Microsatellite analysis of the silkworm strains (Bombyx mori): high variability and potential markers for strain identification

The silkworm (Bombyx mori) is of great economic value from an industrial perspective. Casual discrimination and accumulation of genetic information from a diverse variety of silkworms are essential for practical utilization and longterm conservation. In this study, a total of 54 silkworm strains preserved in Korea were typed for nine polymorphic microsatellite loci. We determined per-locus numbers of alleles ranging from 3 to 17 with an average value of 7.5, per-locus observed heterozygosity ranging from 0.04 to 0.98, and per-locus polymorphic information content (PIC) ranging from 0.06 to 0.86, thereby indicating that some of these loci are profoundly variable. Phylogenetic analysis using the nine concatenated microsatellite loci showed no clustering on the basis of known strain characteristics and origin. A total of 17 strain-specific apomorphic alleles, which discriminate 14 among 54 silkworm strains, were obtained from eight loci. These strain-specific alleles can, therefore, be casually utilized to discriminate between applicable strains, without any further typing of other loci. Furthermore, a substantial number of homozygote strains, represented by 24 among 67 alleles in nine loci, were also detected. These results collectively implicate silkworm microsatellite DNA as useful and potentially important molecular markers for the eventual discrimination of silkworm strains, hundreds of which are currently preserved in Korea.     

Strain identification in Rhizobium trifolii using DNA restriction analysis, plasmid DNA profiles and intrinsic antibiotic resistances

Abstract Indigenous strains of R. trifolii , originating from different geographical regions, were compared using total DNA restriction analysis, plasmid DNA profiles and intrinsic antibiotic resistance patterns. The results of strain relatedness using 2 of these techniques (DNA restriction and antibiotic resistance patterns) were similar, and indicated that these strains formed a heterogeneous group. A large variation in plasmid DNA profiles was observed among the R. trifolii isolates investigated, indicating heterogeneity of plasmid DNA pools. This variation occurred even in some strains that were shown to be related on the basis of their antibiotic resistance patterns. The application of these techniques to differentiate indigenous R. trifolii strains is discussed in this report.