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材小蠹族(Xyleborini)昆虫又称食菌小蠹,是一类蛀干害虫同时具有经济意义,隶属于鞘翅目(Coleoptera),象虫科(Curculionidae),小蠹亚科(Scolytinae)[1-2]。目前该族包括37属,约1 200种[2-3],中国记载10属42种[4]。材小蠹族类群主要取食木质部中的真菌,堵塞疏导组织,并且自身也携带真菌甚至携带病原体入侵植物并加速植物死亡[5]。Xyleborus glabratus Eichhoff所携带的病原菌,就曾在美国的东南部造成了鳄梨(Persea americana Mill. )的大面积死亡,并且在北美洲该族已经成为最具侵略性和攻击性的小蠹类蛀干害虫[3,6]。
小粒绒盾小蠹(Xyleborinus saxesenii(Ratzeburg,1873)),隶属于材小蠹族(Xyleborini),绒盾小蠹属(Xyleborinus)。殷蕙芬[7]1984年将材小蠹族的32种归在材小蠹属下,1991年材小蠹族分成10个属[8],1837年,小粒绒盾小蠹由Ratzeburg发现并定名为Bostrichus saxesenii[9-11],该种有21个同物异名[12-14],多数学者将其称为小粒材小蠹(Xyleborus saxeseni)[15-16],但此命名法未将材小蠹族分属,本文根据形态学观察和测序结果,将该材小蠹分到绒盾小蠹属下,为小粒绒盾小蠹。
小粒绒盾小蠹主要分布在中国(江西、云南、安徽、浙江、广西、福建、贵州、湖南、吉林、江苏、新疆、西藏、台湾、山西、陕西、四川),朝鲜,日本,越南,加拿大,北美,南美,阿根廷,巴西,智利,厄瓜多尔,巴拉圭,乌拉圭等[11,17-18],已知寄主包括橡胶树(Hevea brasiliensis,(H.B.K)Mull. Arg.),五倍子树( Rhus chinensis ,Mill.),山核桃(Carya cathayensis,Sarg.),苹果(Malus pumila,Mill.),云杉(Picea asperata,Mast.),栎树(Quercus stenophylla,Makino),杨梅(Myrica rubra,(Lour.)S. et Zucc.),柿子(Diospyros kaki,Thunb.),锥栗(Castanea henryi,(Skan)Rehd. et Wils. ),桉属植物(Eucalyptus sp.,L'Hér.),槭属植物(Acer sp.,L.),松属植物( Pinus sp. ,Linn),等[14,16,18-21]。
2022年3月,在崂山地区设置多层漏斗型小蠹诱捕器,悬挂地的林分主要是黑松林,周围也有刺槐等阔叶树。在对崂山区小蠹种类监测过程中,发现小蠹诱捕器内收集到一种小蠹,经形态学和分子生物学鉴定。旨在为小蠹类蛀干害虫快速DNA鉴定以及溯源分析提供基础数据。
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2022年3月,在山东省青岛市崂山区(36.315929° N,120.603775° E)设置小蠹诱捕器诱集昆虫成虫。
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采集到的样本储存于无水乙醇中,使用超景深显微镜(基恩士Kenyence VHX-7000)对小蠹的成虫形态特征进行观察和拍照。
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随机抽取4头小蠹成虫,挑胸部和腹部组织,采用T5 Direct PCR Kit(Animal Tissue)试剂盒,提取个体基因组DNA,置于−20 ℃保存待用。
利用小蠹亚科通用引物S1718和A2411 [22]扩增COⅠ 基因,PCR扩增体系为2 × T5 Direct PCR Kit(Animal Tissue)13 μL,S1718 1 μL,A2411 1 μL,DNA模板2 μL和ddH2O 8 μL共25 μL。PCR反应程序为94 ℃预变性3 min;94 ℃变性45 s,47.2 ℃退火1 min,72 ℃延伸1 min,共进行35次循环;72 ℃延伸10 min;产物置于4 ℃保存。PCR扩增产物送北京擎科生物科技有限公司进行测序。
以COⅠ 基因序列建树:选取本试验的X. saxesenii序列(Genbank No. OP002147.1),来自Genbank的12条序列作为参考序列(表1),落叶松八齿小蠹(Ips subelongatus Motschulsky)(Genbank No. KC411952.1)作为外群,运用MAGA 7[23]最大似然法(ML)构建系统发育树[24-26]。
表 1 小蠹科昆虫相关建树序列信息
Table 1. Sequence information in tree construction of Scolytinae
Genbank登录号
Genbank accession No.物种名称
Species采集地
LocationOP002147.1 小粒绒盾小蠹 X. saxesenii 中国,青岛 China: Qingdao MK643403.1 小粒绒盾小蠹 X. saxesenii 美国,科罗拉多州 USA: Colorado MK643400.1 小粒绒盾小蠹 X. saxesenii 美国,科罗拉多州 USA: Colorado MT991655.1 小粒绒盾小蠹 X. saxesenii 美国 USA MN620034.1 小粒绒盾小蠹 X. saxesenii 东南亚 Southeast Asian MT991654.1 小粒绒盾小蠹 X. saxesenii 美国 USA MW617396.1 纹盾材小蠹 Xyleborinus artestriatus Eichhoff 中国 China MN620020.1 纹盾材小蠹 Xyleborinus artestriatus Eichhoff 东南亚 Southeast Asian MW617443.1 Xyleborinus actospinosus Schaufuss 马达加斯加 Madagascar MW617408.1 Xyleborinus sharpae Hopkins, 马达加斯加 Madagascar MW617405.1 Xyleborinus attenuatus Blandford, 马达加斯加 Madagascar OM422678.1 Xyleborinus andrewesi Blandford, 马达加斯加 Madagascar KC411952.1 落叶松八齿小蠹 Ips subelongatus Motschulsky 中国 China
小粒绒盾小蠹Xyleborinus saxesenii(Ratzeburg,1873)的形态和分子鉴定
Morphology and Molecular Identification of Xyleborinus saxesenii(Ratzeburg,1873)(Coleoptera: Scolytinae)
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摘要:
目的 为明确青岛崂山区小蠹种类,本研究对诱集到的小蠹进行分类鉴定,更好地丰富生物多样性。 方法 采用形态学观察和线粒体细胞色素氧化酶亚基Ⅰ(COⅠ)序列分析的方法进行小蠹种类鉴定。 结果 鉴定崂山区的1种小蠹为小粒绒盾小蠹,隶属于象虫总科、小蠹亚科、绒盾小蠹属,该小蠹的典型特征为鞘翅自斜面前缘开始,沟间部具突起的颗瘤,由前向后略许增大,成为纵列,第2沟间部无颗瘤,与2017年吕坤鉴定的山核桃新害虫——小粒绒盾小蠹描述的形态特征一致。基于COⅠ基因序列的系统发育树(ML树)显示,该小蠹样本与美国和东南亚小粒绒盾小蠹样本序列聚成一支。 结论 本文基于形态学和分子生物学两方面的鉴定,明确该诱集到的小蠹为小粒绒盾小蠹并且该种在山东首次记录发现。此研究结果为小蠹快速DNA鉴定以及溯源分析提供基础数据。 Abstract:Objective To identify the species of Scolytinae in Laoshan, the species of the collected Scolytinae were identified to enrich the biodiversity. Method The morphological characteristics of adults Scolytinae were observed and the sequences of the cytochrome oxidase subunit Ⅰ ( COⅠ ) was analyzed to identify the insect. Result A kind of Scolytinae was identified as Xyleborinus saxesenii in Laoshan, belonged to Curculionidae, Scolytinae, Xyleborinus. The typical characteristics of A were the intersulci, which slightly increased from front to back, had protruding granule in elytron starting from the front edge of the slant, and formed a longitudinal row, but there was no granule in the second intersulcus. It was consistent with the morphological characteristics of one new record species of bark beetle——Xyleborinus saxesenii (Ratzeburg, 1873) by LYU kun, which was clustered with the sample sequences of the American and Southeast Asian beetle. Conclusion Based on both morphological and molecular biological identification, this insect is clearly identified as a Xyleborinus saxesenii and is firstly reported in Shandong. The results of this study provide basic data for the rapid DNA identification of the Scolytinae. -
表 1 小蠹科昆虫相关建树序列信息
Table 1. Sequence information in tree construction of Scolytinae
Genbank登录号
Genbank accession No.物种名称
Species采集地
LocationOP002147.1 小粒绒盾小蠹 X. saxesenii 中国,青岛 China: Qingdao MK643403.1 小粒绒盾小蠹 X. saxesenii 美国,科罗拉多州 USA: Colorado MK643400.1 小粒绒盾小蠹 X. saxesenii 美国,科罗拉多州 USA: Colorado MT991655.1 小粒绒盾小蠹 X. saxesenii 美国 USA MN620034.1 小粒绒盾小蠹 X. saxesenii 东南亚 Southeast Asian MT991654.1 小粒绒盾小蠹 X. saxesenii 美国 USA MW617396.1 纹盾材小蠹 Xyleborinus artestriatus Eichhoff 中国 China MN620020.1 纹盾材小蠹 Xyleborinus artestriatus Eichhoff 东南亚 Southeast Asian MW617443.1 Xyleborinus actospinosus Schaufuss 马达加斯加 Madagascar MW617408.1 Xyleborinus sharpae Hopkins, 马达加斯加 Madagascar MW617405.1 Xyleborinus attenuatus Blandford, 马达加斯加 Madagascar OM422678.1 Xyleborinus andrewesi Blandford, 马达加斯加 Madagascar KC411952.1 落叶松八齿小蠹 Ips subelongatus Motschulsky 中国 China -
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