【作者】 陆亚娟；

【导师】 杜予州；

【作者基本信息】 扬州大学 ， 农业昆虫与害虫防治， 2003， 硕士

【摘要】 美洲斑潜蝇Liriomyza sativae是斑潜蝇属中三大主要害虫之一，被世界上许多国家和地区列为检疫性害虫加以控制。本文对美洲斑潜蝇不同地理种群和不同寄主种群的rDNA-ITS和β-tubulin基因进行了克隆和测序，应用不同的生物信息分析软件(DNAStar(Lasergene 501)和Clustal X1.8 & Mega2.1)分析了美洲斑潜蝇种群之间的遗传分化和亲缘关系。 1、通过软件DNAStar(Lasergene 501)和Clustal X1.8 & Mega2.1对美洲斑潜蝇不同寄主种群rDNA-ITS1、rDNA-ITS2、rDNA-ITS全序列进行分析，结果表明，扬州地区的美洲斑潜蝇已存在寄主种群分化，可分为三大类：嗜好寄主豆科(豇豆)、茄科(番茄)和葫芦科(黄瓜)上的种群为一类，它们之间的遗传分化不明显，遗传相似性和遗传距离分别为99.5～100％(DNAStar软件分析结果)和0.000～0.003(Mega2.1软件分析结果)；非嗜好寄主锦葵科(棉花)上的种群为一类，与嗜好寄主种群的遗传相似性和遗传距离分别为15.8～51.1％和0.697～1.140；取食十字花科(青菜)的美洲斑潜蝇种群介于上述两类群之间。在此三类寄主种群中，取食十字花科的种群与嗜好寄主种群和非嗜好寄主种群之间的遗传相似性分别为79.0～98.3％和16.3～50.6％，遗传距离分别为0.007～0.045和0.727～1.181；说明取食十字花科的种群与嗜好寄主种群之间的亲缘关系近于其与非嗜好寄主种群之间的亲缘关系。 通过相同软件对5.8S rDNA进行序列分析，发现5.8S rDNA序列变异不大，但其分析结果与上述分析的结果相似。嗜好寄主种群之间遗传相似性和遗传距离分别为99.2％和0.000；非嗜好寄主种群与嗜好寄主种群遗传相似性和遗传距离分别为86.3～92.2％和0.068；取食十字花科的美洲斑潜蝇种群与嗜好寄主种群和非嗜好寄主种群之间的遗传相似性分别为96.2～98.3％和90.2％，遗传距离分别为0.022和0.092。 2、通过相同软件对美洲斑潜蝇不同地理种群rDNA-ITS1、rDNA-ITS2、rDNA-ITS全序列和5.8S rDNA进行序列分析可见，美洲斑潜蝇在我国也存在一定的地理分化。根据我们对6个省(区)美洲斑潜蝇种群的研究结果来看，主要分为三大类：广东(广州)、广西(南宁)和贵州(铜仁)的美洲斑潜蝇为第一类，它们之间的遗传分化不明显，遗传相似性和遗传距离分别为96.0～100％和0.000～0.007；江苏(扬州)和浙江(杭州)的美洲斑潜蝇种群为另一类，它们之间的遗传分化也不明显，遗传相似性和遗传陆亚娟:美洲斑潜蝇的种群分化研究距离分别为93 .9一100%和0.00任、0.036;海南(檐县)的美洲斑潜蝇种群为第三类。在此三大类地理种群中，海南(澹县)美洲斑潜蝇种群与江苏(扬州)和浙江(杭州)的美洲斑潜蝇种群的遗传相似性和遗传距离分别为67.小100%和0. 000刁.391，与广东 〔广州)、广西(南宁)和贵州(铜仁)的美洲斑潜蝇种群遗传相似性和遗传距离分别为2b.2一52.3%和0.478一1 .233;由此可见，该地区的种群与江苏(扬州)和浙江(杭州)的种群亲缘关系近于与其它地理种群之间的亲缘关系。 3、通过相同软件对不同寄主和不同地区美洲斑潜蝇的旦.加bulin基因序列进行比较分析发现，该基因序列变异不明显，亲缘关系均大于98%。在寄主种群中，寄主为豆科(更豆)、茄科(番茄)、葫芦科(黄瓜)和十字花科(青菜)的美洲斑潜蝇种群之间的遗传相似性和遗传距离分别为99 .0-99.5%和0.005司.011，寄主为锦葵科(棉花)的美洲斑潜蝇种群与其它寄主种群之间的遗传相似性和遗传距离分别为98.小99.0%和0.011一0.016;在地理种群中，海南(澹县)的美洲斑潜蝇种群与江苏(扬州)和浙江 (杭州)的美洲斑潜蝇种群的遗传相似性和遗传距离分别为99.0~9 9 .5%和0.005，它与广东(广州)、贵州(铜仁)和广西(南宁)的美洲斑潜蝇种群的遗传相似性和遗传距离分别为98.4-99.0%和0.011司.016。 从以上p .tubulin基因序列的分析结果可以看出，与rDNA一ITS序列相比，该基因的保守性较好，不适合于美洲斑潜蝇种下分化的研究。更多还原

【Abstract】 Liriomyza sativae is one of the three main pests in Liriomyza, whitch is ranked in the quanrantine pest in many countries and regions. We clonged and sequenced the rDNA-ITS & -tubulin gene of the populations collected from different districts and the populations feeded on different hosts about L sativae. Through different bio-soft of DNAStar (Lasergene 501) and Clustal X1. 8 & Mega2.1, we compared the identity and diversity of gene sequence between different population of L. sativae.By soft of DNAStar (Lasergene 501) and Clustal X1.8 & Mega2.1, we compared the rDNA-ITS 1, rDNA-ITS2 and rDNA-ITS sequence of the populations feeded on the different hosts. The results show that there are the different host populations of L. sativae in Yangzhou district. The different host populations of L sativae could be divided into three classes. The populations feeded on hobby host--Solanaceae (Lycopersicon esculentum), Leguminosae (Vigna sinensis) and Cucurbitaceae (Cucumis sativus) are the first class, the sequence divergence among them is not evident, the identity and divergence of sequence between them are 99.5-1 00%(by DNAStar) and 0.000-0. 003(by Clustal) respectively; the populations feeded on distasteful host-Malvaceae (Gossypium hirsutum) is the second class, the sequence identity and divergence between the population feeded on host-Malvaceae and the populations feeded on hobby host are 15. 8-5 1.1% and 0.697-1.140 respectively; the population feeded on host-Cruciferae (Brassica chinensis) was the third class, the population feeded on host-Cruciferae was among the the first two class. Among the three classes, the sequence identity between the population feeded on Cruciferae-host and the first class , the second class are 79.0-98.3% and 16.3-50.6% respectively, the sequence divergence are 0.007-0.045 and 0.727-1.181 respectively; it shows that the sequence identity between the population feeded on host-Cruciferae and the first class is more closely than the sequence identity between the population feeded on host-cruciferae and the second class.By the same soft, we analyse the 5.8S rDNA sequence. We find that there is little divergence among the 5.8S rDNA sequence, but the result of the analysis was similar to the results of the above mentioned. The identity and divergence between the populationsfeeded on hobby-host are 99.2% and 0.000 respectively; the identity and divergence between the populations feeded on hobby-host and distasteful-host are 86.3-92.2% and 0.0678 respectively; the sequence identity between the population feeded on host-Cruciferae and the populations feeded on hobby-host , distasteful-host are 96.2-98.3% and 90.2% respectively, the sequence divergence are 0.022 and 0.092 respectively.By the same soft, we compared the rDNA-ITS1, rDNA-ITS2, rDNA-ITS and 5.8S rDNA sequence of the populations collected from different geographic area. The outcome is that there is geographic speciation about L. sativae in our country. According to the results of study about the six province(or district), the populations could be divided into three classes. The first class are the populations collected from Guangdong province (Guangzhou) , Guangxi province (Nanning) and Guizhou province (Tongren), there is little divergence between them, the sequence identity and divegence are 96.0-100% and 0.000-0.007 respectively; the second class are the populations collected from Jiangsu province (Yangzhou) and Zhejiang province (Hangzhou), the sequence identity and divergence are 93.9-100% and 0.000-0.036 respectively; the population collected from Hainan(Danxian) is the third class. Among the three classes, the sequence identity and divergence between the population collected from Hainan(Danxian) and populations collected from Jiangsu province (Yangzhou) , Zhejiang province (Hangzhou) ’s are 67.4-100% and 0.000-0.391 respectively, the sequence identity and divergence between the population collected from Hainan(Danxian) and the first class are 26.2-52.3% and 0.478-1.233 respectively; it shows that the homogenous betwe更多还原