几种斑潜蝇的超微结构及rDNA ITS序列比较研究
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摘要
斑潜蝇Liriomyza是一类世界性分布的害虫，是世界上许多国家蔬菜和花卉生产上最为重要的害虫之一；其中美洲斑潜蝇和南美斑潜蝇是世界上许多国家和地区的重要检疫性害虫。斑潜蝇在我国分布广泛，给我国的蔬菜和花卉生产造成了严重损失。为解决目前我国在斑潜蝇传统分类鉴定上存在的一些问题，本文通过电子显微镜、生化和分子生物学技术，对斑潜蝇属中的美洲斑潜蝇、南美斑潜蝇和番茄斑潜蝇的分类鉴定进行了研究，并对我国美洲斑潜蝇的种下分化作了分析。
1、通过对美洲斑潜蝇、南美斑潜蝇和番茄斑潜蝇超微结构的观察，发现前翅前缘上的刺毛形状和排列方式、雄成虫的阳茎在这三种斑潜蝇之间存在明显差异。通过对幼虫后气门突和卵的超微结构观察，发现幼虫后气门作为分类特征不如在光学显微镜中明显；而不同斑潜蝇的卵在电镜下的形态却有明显差异。通过研究发现，不同地区美洲斑潜蝇雌虫产卵器形态特征和表面饰纹具有一定的差异。因此我们认为，斑潜蝇前翅前缘上的刺毛特征和卵的超微结构特征可作为斑潜蝇的种类鉴别依据；雌成虫产卵器的超微结构特征可以作美洲斑潜蝇种下分化的判别依据之一。
2、美洲斑潜蝇、南美斑潜蝇和番茄斑潜蝇酯酶的聚丙烯酰胺凝胶电泳结果存在着显著的差异，其中美洲斑潜蝇酯酶有5条酶带，南美斑潜蝇有3条酶带，番茄斑潜蝇有6酶带。
3、对美洲斑潜蝇、南美斑潜蝇和豌豆彩潜蝇的rDNA ITS序列进行了分析比较，发现美洲斑潜蝇和南美斑潜蝇的亲缘关系很近，遗传同源性高达98.3％；美洲斑潜蝇和豌豆彩潜蝇的遗传同源性为89.3％，南美斑潜蝇和豌豆彩潜蝇的遗传同源性为89.6％。
4、对采集自江苏(扬州)、浙江、广西和海南的美洲斑潜蝇的rDNA ITS序列作了比较分析，结果表明他们的亲缘关系依次为：江苏和浙江的遗传同源性为99.1％，江苏和广西的遗传同源性为93.3％，江苏和海南的遗传同源性为84.8％。这些分析结果表明，不同的地理种群，其遗传分化随地理差异的增大而增大。
5、通过对不同寄主上的美洲斑潜蝇的rDNA ITS序列比较分析发现，不同寄主上的美洲斑潜蝇种群具有一定的遗传分化，但在嗜好寄主豇豆、番茄和西葫芦上的亲缘关系较近，其中番茄上的与豇豆上的遗传同源性为99.7％，番茄上的与西葫芦上的为95.3％；而在非嗜好寄主棉花和油菜上的亲缘关系也比较近，其遗传同源性为94.5％。
The vegetable leafminer, Liriomyza that distributes all over the world is a category of most important pests of vegetable and flowers in many countries. In Liriomyza, L. sativae, L. huidobrensis are ultimately objects subject to quanrantine in many countries and areas. Leafminers distribute widely in our country, and make great damage in vegetable and flowers production. To solve problem that existed in methods of classifying and identifying leafminers in our country, and provide scientific basis for preventing and curing the leafminers. Through electron microscope, biochemical and molecular biological technology, we studied the classification of L. sativae, L. huidobrensis and L. brynosis, and we also researched the differentiation of host and geographic populations of L sativae in China.
Through observing microtexture of L. sativae, L. huidobrensis and L. brynosis, we found there were obvious differentiation in furca and its form of arrangement in wings costa and aedeagus of male adult of three leafminers; through observing poststigma of larvae and figure of eggs, we found that the function of classification identification was not as same as in optical microscope, but the figures of eggs were obviously different in electron microscope. We also found there was difference in ovipositor's surface patterns and the figures of geographic populations of L. Sativae. So we think that the ultrastucture characters of wing costa and eggs stings could be seemed as important basis for identifying different species of Liriomyza; the ultrastucture characters of ovipositor could be seemed as basis for identifying population divergence of Liriomyza sativae.
There were obviously differentiations in outcome of PAGE of L sativae, L huidobensis and L. bynosis. In Esterase Isozymes spectrum, L. sativae had 5 strips, L. huidobensis had 3 strips and L. bynosis had 6 strips.
We determined and compared the rDNA ITS sequence of I. sativae, L. huidobrensis and Chhramatomyia harticola, we found that it was closed in relationship of affimity of L. sativae and L. huidobrensis, the homogenous was 98.3%; and the homogenous of L. sativae and Chhramatomyia harticola was 89.3%, the homogenous of L. huidobrensis and Chhramatomyia harticola is 89.6%.
Comparing the rDNA ITS sequence of L. sativae collected from Jiangsu privince (Yanghzou), Zhejiang province, Guangxi province and Hainan province, the outcome indicated that they were closed orderly in relationship of affimity: homogenous of Jiangsu and Zhejiang was 99.1%, homogenous of Jiangsu and Guangxi was 93.3% and homogenous of Jiangsu and Hainan was 84.8%. All of these illustrated that hereditary divergence degree of geographic populations changed with the divergence degree of the geographic conditions.
The rDNA ITS sequence of L. sativae collected from different hosts also were compared. It could be found that it is closed in relationship of affimity of hobby hosts-cowpea; tomato and edible gourd (homogenous of tomato and cowpea was 99.7%, homogenous of tomato and edible gourd was 95.3%) and same in non-hobby hosts-cotton and Chinese cabbage (homogenous was 94.5%) .
1、通过对美洲斑潜蝇、南美斑潜蝇和番茄斑潜蝇超微结构的观察，发现前翅前缘上的刺毛形状和排列方式、雄成虫的阳茎在这三种斑潜蝇之间存在明显差异。通过对幼虫后气门突和卵的超微结构观察，发现幼虫后气门作为分类特征不如在光学显微镜中明显；而不同斑潜蝇的卵在电镜下的形态却有明显差异。通过研究发现，不同地区美洲斑潜蝇雌虫产卵器形态特征和表面饰纹具有一定的差异。因此我们认为，斑潜蝇前翅前缘上的刺毛特征和卵的超微结构特征可作为斑潜蝇的种类鉴别依据；雌成虫产卵器的超微结构特征可以作美洲斑潜蝇种下分化的判别依据之一。
2、美洲斑潜蝇、南美斑潜蝇和番茄斑潜蝇酯酶的聚丙烯酰胺凝胶电泳结果存在着显著的差异，其中美洲斑潜蝇酯酶有5条酶带，南美斑潜蝇有3条酶带，番茄斑潜蝇有6酶带。
3、对美洲斑潜蝇、南美斑潜蝇和豌豆彩潜蝇的rDNA ITS序列进行了分析比较，发现美洲斑潜蝇和南美斑潜蝇的亲缘关系很近，遗传同源性高达98.3％；美洲斑潜蝇和豌豆彩潜蝇的遗传同源性为89.3％，南美斑潜蝇和豌豆彩潜蝇的遗传同源性为89.6％。
4、对采集自江苏(扬州)、浙江、广西和海南的美洲斑潜蝇的rDNA ITS序列作了比较分析，结果表明他们的亲缘关系依次为：江苏和浙江的遗传同源性为99.1％，江苏和广西的遗传同源性为93.3％，江苏和海南的遗传同源性为84.8％。这些分析结果表明，不同的地理种群，其遗传分化随地理差异的增大而增大。
5、通过对不同寄主上的美洲斑潜蝇的rDNA ITS序列比较分析发现，不同寄主上的美洲斑潜蝇种群具有一定的遗传分化，但在嗜好寄主豇豆、番茄和西葫芦上的亲缘关系较近，其中番茄上的与豇豆上的遗传同源性为99.7％，番茄上的与西葫芦上的为95.3％；而在非嗜好寄主棉花和油菜上的亲缘关系也比较近，其遗传同源性为94.5％。
The vegetable leafminer, Liriomyza that distributes all over the world is a category of most important pests of vegetable and flowers in many countries. In Liriomyza, L. sativae, L. huidobrensis are ultimately objects subject to quanrantine in many countries and areas. Leafminers distribute widely in our country, and make great damage in vegetable and flowers production. To solve problem that existed in methods of classifying and identifying leafminers in our country, and provide scientific basis for preventing and curing the leafminers. Through electron microscope, biochemical and molecular biological technology, we studied the classification of L. sativae, L. huidobrensis and L. brynosis, and we also researched the differentiation of host and geographic populations of L sativae in China.
Through observing microtexture of L. sativae, L. huidobrensis and L. brynosis, we found there were obvious differentiation in furca and its form of arrangement in wings costa and aedeagus of male adult of three leafminers; through observing poststigma of larvae and figure of eggs, we found that the function of classification identification was not as same as in optical microscope, but the figures of eggs were obviously different in electron microscope. We also found there was difference in ovipositor's surface patterns and the figures of geographic populations of L. Sativae. So we think that the ultrastucture characters of wing costa and eggs stings could be seemed as important basis for identifying different species of Liriomyza; the ultrastucture characters of ovipositor could be seemed as basis for identifying population divergence of Liriomyza sativae.
There were obviously differentiations in outcome of PAGE of L sativae, L huidobensis and L. bynosis. In Esterase Isozymes spectrum, L. sativae had 5 strips, L. huidobensis had 3 strips and L. bynosis had 6 strips.
We determined and compared the rDNA ITS sequence of I. sativae, L. huidobrensis and Chhramatomyia harticola, we found that it was closed in relationship of affimity of L. sativae and L. huidobrensis, the homogenous was 98.3%; and the homogenous of L. sativae and Chhramatomyia harticola was 89.3%, the homogenous of L. huidobrensis and Chhramatomyia harticola is 89.6%.
Comparing the rDNA ITS sequence of L. sativae collected from Jiangsu privince (Yanghzou), Zhejiang province, Guangxi province and Hainan province, the outcome indicated that they were closed orderly in relationship of affimity: homogenous of Jiangsu and Zhejiang was 99.1%, homogenous of Jiangsu and Guangxi was 93.3% and homogenous of Jiangsu and Hainan was 84.8%. All of these illustrated that hereditary divergence degree of geographic populations changed with the divergence degree of the geographic conditions.
The rDNA ITS sequence of L. sativae collected from different hosts also were compared. It could be found that it is closed in relationship of affimity of hobby hosts-cowpea; tomato and edible gourd (homogenous of tomato and cowpea was 99.7%, homogenous of tomato and edible gourd was 95.3%) and same in non-hobby hosts-cotton and Chinese cabbage (homogenous was 94.5%) .
引文
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