城市绿化树种槐树之蚧害调查及其遗传差异研究
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摘要
槐树(Sophora japonica L.),适应性强,易成活,特别在吸收污染物、净化大气等方面具有独特性,成为城市绿化优选树种之一,被广泛种植用于街道绿化。但是,这种良好的绿植生态环境,也给众多有害生物提供了极好的定殖场所,其中蚧虫成为了槐树的优势危害种群,同时它极易随种苗、大树的调运,在空间上远距离传播,侵入到新的生态体系引起广泛危害,给农林生产安全造成极大威胁。因此,本实验对太原市三条主要交通路段上的槐树进行抽样分析,寻找不同的蚧虫种类,并对不同生态位上的蚧虫数量、种类进行统计学分析,从槐树与蚧虫的关系入手对蚧虫选择寄主、与共栖寄生物协同作用、生物生态学等方面进行系统的研究,以提出科学的防控措施。同时通过对蚧虫从遗传差异方面的分子生物学研究,建立快速准确的鉴定技术,以加强和保障植物检疫措施。主要内容如下：
第一：实验对太原市漪汾街(幼龄槐树比较集中)、南内环(中龄槐树比较集中)、晋祠路(老龄槐树比较集中)上的槐树进行抽样分析,寻找不同的蚧虫种类,并对不同生态位上的蚧虫数量、种类进行统计学分析,之后进行种类鉴定,搞清楚我市槐树主要受哪种蚧虫的侵害,汽车尾气、灰尘等污染对蚧虫的分布是否有影响。根据我们的分析,我们已发现枣大球蚧(Eulecanium gigantea (Shinji))和皱大球坚蚧(Eulecanium kuwanai Kanda)是危害我省槐树的主要种类。
第二：实验以五种危害槐树较严重的蚧虫为材料,分别用SDS法(十二烷基硫酸钠)、CTAB法(十六烷基三乙基溴化铵)、TES法(酚-氯仿)、NaC1法(氯化钠)和TNES法5种提取方法,对单只蚧虫进行全基因组DNA提取。结果表明,5种提取方法都能够获得基因组DNA,但是相比较而言,CTAB法和TES法所提取的DNA质量明显优于SDS法、NaCl法和TNES法。因此认为,CTAB法和TES法比较适合实验室提取蚧虫基因组DNA。
第三：实验以五种危害槐树较严重的蚧虫为材料,采用基因扩增技术,对五种蚧虫的COI-COⅡ基因进行PCR扩增和测序,并比对其序列设计各自的特异性引物。结果表明,基于PCR扩增及直接测序的分子技术可以达到准确、快速鉴定五种蚧虫的目的。
Sophora japonica L easy to survive, particularly in the absorption of pollutants, air purification and so unique, as one of city greening tree species selection, is widely grown for street greening. However, this favorable environment green plants, but also to many pest colonization provides an excellent venue which has become a locust tree scale insect populations of the advantages of harm, and it easily with the seedlings and trees transporting, Long-distance spread in space, invade a new ecological system caused widespread harm to the agricultural and forestry production caused a great threat to security.
First, this experiment in Taiyuan Yi Fen Street (more concentrated in young Ash), South inner (more concentrated in the age of Ash), Jinci Road (old acacia trees are concentrated) sample survey on the acacia trees, looking for different The scale insect species, and different niche on the number of scale insect, the type used for statistical analysis, after species identification, find out what the city is mainly affected by acacia trees against scale insects, vehicle exhaust, dust and other pollution Kuwana Would affect the distribution of insects. Finally, rapid, accurate and effective quarantine methods. According to our investigation, we have found great dates the Scale Eulecanium gigantea (Shinji), and fold the big ball firmly Eulecanium kuwanai Kanda locust tree in our province is the main type of hazard.
Two, Genomic DNA were extracted from five scale insect by using the sodium dodecyl Sulfate (SDS) method, the cetylt ethylammonium bromide (CTAB) method, Phenol-chloroform (TES) method, the sodium chloride (NaC1)method and TNES method respectively. The results show that the five methods were useful in extraction of genomic DNA of scale insects. But comparatively speaking, the quality of DNA samples isolated by using the CTAB method and the TES method was obviously better than that by using the other three methods. So that, our laboratory think that the CTAB and the TES method were more effective and practical method for the genomic DNA extraction of scale insect.
Three, The mitochondrial COI-COII sequences were of five scale insects were were amplified by using PCR technique to analyze their sequence variation and then design their respective specific primer sequence. The results show that, based on the molecular techniques of PCR amplification and direct sequencing can achieve accurate,rapid identification of five scale insect.
第一：实验对太原市漪汾街(幼龄槐树比较集中)、南内环(中龄槐树比较集中)、晋祠路(老龄槐树比较集中)上的槐树进行抽样分析,寻找不同的蚧虫种类,并对不同生态位上的蚧虫数量、种类进行统计学分析,之后进行种类鉴定,搞清楚我市槐树主要受哪种蚧虫的侵害,汽车尾气、灰尘等污染对蚧虫的分布是否有影响。根据我们的分析,我们已发现枣大球蚧(Eulecanium gigantea (Shinji))和皱大球坚蚧(Eulecanium kuwanai Kanda)是危害我省槐树的主要种类。
第二：实验以五种危害槐树较严重的蚧虫为材料,分别用SDS法(十二烷基硫酸钠)、CTAB法(十六烷基三乙基溴化铵)、TES法(酚-氯仿)、NaC1法(氯化钠)和TNES法5种提取方法,对单只蚧虫进行全基因组DNA提取。结果表明,5种提取方法都能够获得基因组DNA,但是相比较而言,CTAB法和TES法所提取的DNA质量明显优于SDS法、NaCl法和TNES法。因此认为,CTAB法和TES法比较适合实验室提取蚧虫基因组DNA。
第三：实验以五种危害槐树较严重的蚧虫为材料,采用基因扩增技术,对五种蚧虫的COI-COⅡ基因进行PCR扩增和测序,并比对其序列设计各自的特异性引物。结果表明,基于PCR扩增及直接测序的分子技术可以达到准确、快速鉴定五种蚧虫的目的。
Sophora japonica L easy to survive, particularly in the absorption of pollutants, air purification and so unique, as one of city greening tree species selection, is widely grown for street greening. However, this favorable environment green plants, but also to many pest colonization provides an excellent venue which has become a locust tree scale insect populations of the advantages of harm, and it easily with the seedlings and trees transporting, Long-distance spread in space, invade a new ecological system caused widespread harm to the agricultural and forestry production caused a great threat to security.
First, this experiment in Taiyuan Yi Fen Street (more concentrated in young Ash), South inner (more concentrated in the age of Ash), Jinci Road (old acacia trees are concentrated) sample survey on the acacia trees, looking for different The scale insect species, and different niche on the number of scale insect, the type used for statistical analysis, after species identification, find out what the city is mainly affected by acacia trees against scale insects, vehicle exhaust, dust and other pollution Kuwana Would affect the distribution of insects. Finally, rapid, accurate and effective quarantine methods. According to our investigation, we have found great dates the Scale Eulecanium gigantea (Shinji), and fold the big ball firmly Eulecanium kuwanai Kanda locust tree in our province is the main type of hazard.
Two, Genomic DNA were extracted from five scale insect by using the sodium dodecyl Sulfate (SDS) method, the cetylt ethylammonium bromide (CTAB) method, Phenol-chloroform (TES) method, the sodium chloride (NaC1)method and TNES method respectively. The results show that the five methods were useful in extraction of genomic DNA of scale insects. But comparatively speaking, the quality of DNA samples isolated by using the CTAB method and the TES method was obviously better than that by using the other three methods. So that, our laboratory think that the CTAB and the TES method were more effective and practical method for the genomic DNA extraction of scale insect.
Three, The mitochondrial COI-COII sequences were of five scale insects were were amplified by using PCR technique to analyze their sequence variation and then design their respective specific primer sequence. The results show that, based on the molecular techniques of PCR amplification and direct sequencing can achieve accurate,rapid identification of five scale insect.
引文
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[3]刘贵峰,刘玉平.国槐蚧虫种类鉴定、危害及防治的研究[J].内蒙古民族大学学报(自然科学版),2004,54-57
[4]汤枋德.中国蚧科[M].山西,山西高校联合出版社,1991,1-377
[5]汤枋德.中国园林主要蚧虫第一卷[M].沈阳,沈阳出版社,1977,1-259
[6]谢映平.山西林果蚧虫[M].北京,中国林业出版社,1998,1-147
[7]梁忆冰,詹国平,徐亮等.进境花卉有害生物风险初步分析[J].植物检疫,1999,13(1),17-22
[8]胡学难.检疫性蚧虫研究的现状与对策[J].植物检疫,1998,27,84-91
[9]顾忠盈,邰明,刁彩华.江苏出口盆景蚧虫种类及其检疫措施.植物检疫[J],1998,13,154-156
[10]武三安,俞思佳,付怀军.北京发现的园林新害虫-仙人掌根粉蚧[J].植物检疫,2001,15(3),158-159
[11]薛皎亮,刘红霞,谢映平.城市空气中铅在国槐树体内的积累[J].中国环境科学,2000,20(6),536-539
[12]薛皎亮.国槐对城市街道空气中铅污染物吸收作用研究初报[J].山西林业科技,1995,4,47-48
[13]原贵生.国槐对空气硫污染物吸收与净化作用研究[J].林业科技通讯,1997,2.25-26
[14]蒋冰海.美育学导论[M].上海,上海人民出版社,1990,1-229
[15]张春妮,戴武,花保祯.中国北方果树蚧虫为害及天敌利用分析[J].西北农业学报,2005,14(2),96-100
[16]齐晓丰.北京地区蚧虫区系的研究[D].北京林业大学,硕士学位论文,2008,10-12
[17]高宝嘉,崔巍,李俊英.华北地区观赏树种蚜虫为害及种类识别[J].河北林学院学报,1996,11(1),56-61
[18]王珊珊.北京地区温室蚧虫种类调查及蜡蚧轮枝菌对长尾粉蚧的毒力测定 [D].北京林业大学,硕士学位论文,2010,8-13
[19]高日霞,陈景耀.中国果树病虫原色图谱[M].北京,中国农业出版社,2011,1-31l
[20]韩乃勇,杨莉,沙尼亚.苹果球蚧的发生与防治[J].新疆农业科技,2006,5,32-33
[21]吴金亮,张富和,周恩义等.朝鲜球坚蜡蚧在苹果树上的发生与防治[J].西北园艺(果树专刊),2005,l,23-24
[22]汤祊德.我国蚧虫研究的历史、现状和展望[J].武夷科学,2001,17,82-86
[23]陈英,李学骝泽.蚧虫研究进展[J].丽水农业科技,1989,1,7-14
[24]谢映平,郑乐怡.瘤坚大球蚧蜡泌物的超微结构与红外光谱特征昆虫学报[M].2001,44(4),408-415
[25]谢映平,郑乐怡.朝鲜毛球蚧蜡泌物的超微形态与红外光谱特征[J].昆虫学报,2002,45(3),329-335
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