柑橘黄龙病株不同部位内生细菌群落结构的多样性  

The community structure of endophytic bacteria in different parts of huanglongbing-affected citrus plants

作　　者：刘波[1] 郑雪芳[1] 孙大光[1,2] 阮传清[1] 范国成[3] 段永平[4] 

机构地区：[1]福建省农业科学院农业生物资源研究所,福州350003 [2]福建师范大学生命科学学院,福州350003 [3]福建省农业科学院果树研究所,福州350003 [4]美国农业部佛罗里达园艺试验室,弗罗里达34945

出　　处：《生态学报》2011年第24期7325-7342,共18页Acta Ecologica Sinica

基　　金：中美合作项目(柑橘木虱生防菌与抗虫柑橘品种研究);福建省财政专项-福建省农业科学院科技创新团队建设基金(STIF-Y03)

摘　　要：采用PCC(Pearson correlation coefficient)、系统聚类和多样性指数研究了柑橘黄龙病(citrus huanglongbing,HLB)植株不同部位叶片、枝条及其根部内生细菌的相互关系及其群落结构多样性。结果表明,从供试柑橘中分离得到26株内生细菌(叶片10株,枝条14株,根部2株)隶属于10个属19个种。Nested-PCR结果显示,柑橘植株不同空间叶片中,黄龙病病原的阳性检出率为58.3%。用LSD法比较显示,柑橘内生细菌在不同器官的分布量大小依次为根>叶>枝条。上、中、下不同部位叶片和枝条内生细菌分布量差异不显著,而东、西、南、北不同方位的叶片和枝条内生细菌分布量差异显著(P<5%)。柑橘内生细菌之间的PCC分析表明,Bacillus pumilus和Bacillus sp.与柑橘黄龙病病原菌呈显著负相关,Bacillus sp.和Bacillus pumilus与Bacillussubtilis呈显著正相关。Bacillus brevis和Escherichia hermannii与Bacillus vesiculariss存在显著正相关。对柑橘不同器官内生细菌的PCC分析表明,同种器官的内生细菌之间呈正相关,不同器官的内生细菌之间呈负相关。以内生细菌在各器官的分布量为指标分析不同器官之间的相关性,结果表明,叶片与枝条呈正相关,PCC为0.55,叶片与根部、枝条与根部均呈负相关,PCC分别为-0.19和-0.13。16S rDNA序列聚类分析表明,柑橘内生细菌可分为两大类,第Ⅰ大类为革兰氏阳性菌,第Ⅱ大类为革兰氏阴性菌。第Ⅰ大类又可以分为芽孢杆菌属和短小杆菌属两个亚类,第Ⅱ大类也可以分两个亚类。对柑橘黄龙病病原与叶片内生细菌进行聚类分析,可以分为三类,第Ⅰ类特征为内生细菌存在所有部位的叶片;第Ⅱ类特征为内生细菌与黄龙病病原菌存在负相关性;第Ⅲ类特征为内生细菌只存在特定部位叶片。当马氏距离为27.23时,可将柑橘黄龙病内生细菌群落结构聚为3类,第Ⅰ类特点是在不同部位叶片均有分布,为完全分布类型,且�The analyses methods of Pearson correlation coefficient（PCC）,hierarchical cluster analysis and diversity index were used to study the relevance between citrus huanglongbing（HLB） and the endophytic bacteria in different branches and leaves as well as roots of huanglongbing（HLB）-affected citrus trees.The results showed that 26 isolates（10 isolates from leaves,14 isolates from branches and 2 isolates from roots） from HLB-affected citrus plants belonged to 19 bacterial species in 10 Genus.Among the 12 tested leaf samples,58.3% of them were positive for the HLB pathogen using Nested-PCR.The numbers of endophytic bacteria in the different parts of citrus plants ranked as in citrus root in leaves in sticks using statistical LSD（least significant difference）.The numbers of endophytic bacteria in leaves with different orientations were significant different but little differenct were found among upper parts,middle parts and lower parts.Among the bacterial populations,Bacillus pumilus and Bacillus sp.were significantly negatively correlated to HLB.bacterium;Bacillus sp.and Bacillus pumilus was positively correlated to Bacillus subtilis,while Bacillus brevis and Escherichia hermannii was positively correlated to Bacillus vesicularis.Furthermore,there was no correlation between Bacillus subtilis and Bacillus brevis based on PCC analysis.The correlations among the bacteria from the same tissue such as the leaves,sticks and roots were significantly positive,while the correlations among the bacteria from the different tissues were negative.The correlation coefficient of endophytic bacteria between the leaves and the sticks was over 0.55（P0.05）.Based on clustering analysis of 16S rRNA gene sequences,citrus endophytic bacteria was divided into two groups：Gram-positive and Gram-negative.The correlations between HLB bacterium and other bacteria were divided into 3 groups.The bacteria in group Ⅰ distributed in all leaves;and the bacterias in the group Ⅱ were negatively corrected to HLB bacterium while i

关 键 词：柑橘黄龙病 内生细菌 多样性 群落结构 

分 类 号：S436.66[农业科学—农业昆虫与害虫防治]