黄山马尾松林天牛及携带线虫种类初步调查

王立超; 苏胜荣; 陈凤毛; 董晓燕; 田成连; 王洋

doi:10.12302/j.issn.1000-2006.202106041

黄山马尾松林天牛及携带线虫种类初步调查

王立超, 苏胜荣, 陈凤毛, 董晓燕, 田成连, 王洋

南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4) : 29-35.

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南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (4) : 29-35. DOI: 10.12302/j.issn.1000-2006.202106041

专题报道Ⅰ：松材线虫病致病机理与病害防治（执行主编　叶建仁）

黄山马尾松林天牛及携带线虫种类初步调查

王立超 ¹ ,
苏胜荣 ²^,^△ ,
陈凤毛 ¹^,^* ,
董晓燕 ¹ ,
田成连 ³ ,
王洋 ¹

作者信息 +

Preliminary investigations of vector beetle and nematode species in Pinus massoniana forest in Huangshan City

Author information +

文章历史 +

摘要

【目的】松材线虫病是我国危害最大的森林病害,松树一旦感病便迅速死亡。调查黄山地区马尾松林内天牛种类以及天牛携带线虫的变化规律,为黄山市松材线虫病的防控工作提供理论支持。【方法】在安徽省黄山市博村林场通过野外和室内收集天牛,并对天牛进行整姿、鉴定,记录天牛种类和数量。为确定可以携带线虫的媒介天牛,使用贝尔曼漏斗法对可以危害松树且有活力的天牛进行单头天牛线虫分离实验,12 h后,用15 mL离心管接取漏斗下端的天牛浸泡液,将浸泡液放在显微镜下观察是否含有线虫。根据携带线虫的天牛数量占天牛总数的比例计算天牛总体携带线虫比例;为鉴定天牛携带线虫种类,先通过室内真菌培养、观察线虫形态学特征对线虫进行初步鉴定,再使用松材线虫分子检测技术确定天牛携带的线虫是否为松材线虫,对于其他寄生线虫,通过28S rDNA基因进行扩增、测序、比对以及构建系统进化树进行分类鉴定。此外,统计不同时间携带线虫的天牛数量以及天牛携带线虫数量的变化。【结果】在黄山市博村林场共收集到4个亚科12个属的14种天牛,通过线虫分离实验确定可以携带线虫的天牛主要有2种:松墨天牛(Monochamus alternatus)和小灰长角天牛(Acanthocinus griseus),2种天牛整体线虫携带率分别为45.8%和13.6%,通过形态学观察松墨天牛和小灰长角天牛均可携带两种不同形态的扩散型线虫,一种是尖尾形线虫,另一种则是圆尾形线虫。尖尾形线虫通过形态学和松材线虫分子检测鉴定被确定为松材线虫(Bursaphelenchus xylophilus),而圆尾形线虫则通过形态学和系统发育树结果被确定为Contortylenchus sp.类线虫。6月19日至7月23日每周线虫分离试验结果表明,松墨天牛雌成虫携带线虫的比例依次为76.5%、66.0%、52.9%、34.7%和10.0%;松墨天牛雄成虫携带线虫的比例依次为76.5%、60.0%、50.0%、30.8%和14.7%,松墨天牛雌成虫携带线虫的数量依次为1 018.0、530.1、202.5、200.4和80.0条/头,松墨天牛雄成虫携带线虫的数量依次为495.1、68.3、74.9、8.2和3.8条/头。【结论】黄山市博村林场马尾松林内共收集到14种天牛,其中松墨天牛和小灰长角天牛都可以携带松材线虫和Contortylenchus sp.类线虫,但以松墨天牛为主,松墨天牛携带线虫的数量和比例随携带时间推移逐渐减少。

Abstract

【Objective】The pine wilt disease is one of the serious forestry diseases in China, and causes rapid death in pine trees. To provide a theoretical support for the prevention and control of pine wood disease in Huangshan City, an investigation on the species of longicorn beetles and related nematodes was carried out in Pinus massoniana forest. 【Method】Longicorn beetles were collected indoors and outdoors from the Bocun forest farm. The beetles were fixed and identified, and their species and number were recorded. To confirm the presence of nematode vectors, Baermann funnel technique was used to extract the nematodes from beetles, which are harmful to the vitality of pine trees. After soaking the beetles for 12 h, a 15 mL centrifuge tube was used to place the soaking solution at the end of the funnel, which was then observed under a microscope to determine the presence or absence of nematodes. The ratio of beetles that carry nematodes was calculated by determining the ratio of nematode-containing beetles to the total number. To identify the species of nematodes carried by beetles, they were first cultured with fungi and preliminarily identified by observing their morphological characteristics. The molecular identification technique of Bursaphelenchus xylophilus was used to determine if the nematode was B. xylophilus. For the other parasitic nematodes, the 28S rDNA gene of nematodes was amplified, sequenced, blasted, and the phylogenetic tree was constructed for identification. Besides, the ratio and the number of nematodes carried by beetles at different times were counted. 【Result】 Total 14 longicorn beetles belonging to 12 genera and four subfamilies were collected. Two species of beetles Monochamus alternatus and Acanthocinus griseus were associated with nematodes, and their overall nematode carrying rates were 45.8% and 13.6%, respectively. M. alternatus and A. griseus both carry two types of dispersal nematodes with different morphological characteristics, one with a pointed tail and the other with a rounded tail. Results of the morphological and molecular analysis revealed the nematode with the pointed tail as B. xylophilus, while the other as Contortylenchus sp. During the period from June 19 to July 23, the weekly nematode isolation test results showed that the nematodes carried by female adults of M. alternatus were 76.5%, 66.0%, 52.9%, 34.7% and 10.0%, respectively; the proportion of nematodes carried by male adults of M. alternatus was 76.5%, 60.0%, 50.0%, 30.8% and 14.7%. The average number of nematodes carried by per individual female adult of M. alternatus was 1 018.0, 530.1, 202.5, 200.4 and 80.0, and the average number of nematodes carried by per individual male adult was 495.1, 68.3, 74.9, 8.2 and 3.8. 【Conclusion】 A total of 14 longicorn beetles were recorded in the masson pine forest in Bocun forest farm, Huangshan City. Among them, M. alternatus and A. griseus can carry B. xylophilus and Convertylenchus sp., the former being the main strain. The number and proportion of M. alternatus carrying nematodes gradually decreased with time.

导出引用

王立超, 苏胜荣, 陈凤毛, 等. 黄山马尾松林天牛及携带线虫种类初步调查[J]. 南京林业大学学报（自然科学版）. 2022, 46(4): 29-35 https://doi.org/10.12302/j.issn.1000-2006.202106041

WANG Lichao, SU Shengrong, CHEN Fengmao, et al. Preliminary investigations of vector beetle and nematode species in Pinus massoniana forest in Huangshan City[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2022, 46(4): 29-35 https://doi.org/10.12302/j.issn.1000-2006.202106041

中图分类号： S763

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