柑橘植株根系微生物生态学特性的研究讲解.docVIP

柑橘植株根系微生物生态学特性的研究 郑雪芳、刘波*、阮传清、林营志、肖荣风、朱育菁、范国成、蔡子坚 （福建省农业科学院柑橘黄龙病研究中心） 摘要：从福建福州市北峰的柑橘园基地采集了8份不同发病状态柑橘根系土壤，用常规的NA培养基进行细菌分离，利用磷脂脂肪酸（PLFA）对分离的细菌进行鉴定。结果表明：不同品种、不同发病状态的柑橘根系土壤共分离出40种微生物，其中福橘品种黄龙病发病株和健株的根系土壤共分离出13种微生物，脐橙黄龙病发病株和健株的根系土壤共分离出9种微生物，雪柑黄龙病发病株和健株的根系土壤共分离出6种微生物。福橘品种黄龙病株和健株根系土壤均分布有球形节杆菌，但在黄龙病株的根系土中分布量较大。脐橙黄龙病株根系土壤微生物群落较健株根系土壤丰富。比较雪柑黄龙病株和健株根系土壤微生物分布量，我们发现雪柑黄龙病株根系土壤微生物分布量明显比健株根系土壤微生物分布量大。 关键词：柑橘；根系微生物；生态学特性；PLFAs Study on the ecology of citrus rhizospheric microbial community ZHENG Xue-Fang, LIU Bo, RUAN Chuan-Qing, LIN Ying-Zhi, XIAO Rong-Feng, ZHU Yu-Jing, FAN Guo-Cheng, CAI Zi-Jian (Citrus Huanglongbing Center of Fujian Academy of Agricultural Sciences) Abstract: Eight samples of rhizospheric soil were collected from the Beifeng Experimental Citrus Orchard in Fuzhou City, Fujian Province of southern China. The rhizospheric bacteria in the soil samples were isolated with NA medium and identified with PLFAs. As the result, 40 bacteria species were isolated from all the samples. Among these bacteria, 13 species were isolated from the rhizospheric soil samples of huanglongbing infected and healthy plants of citrus variety “Fuju”, 9 species from the samples of huanglongbing infected and healthy plants of citrus variety “Qicheng”, and 6 species from the samples of variety “Xuegan”. Among rhizospheric bacteria of variety “Fuju”, Arthrobacter globiformils were detected with higher plenty for huanglongbing infected plants than the healty ones. For varieties of “Qicheng” and “Xuegan”, the rhizospheric bacteria were more plentiful for the huanglongbing infected plants than for the healthy ones. Key words: citrus, rhizospheric bacteria, ecology, PLFAs 黄龙病（Citrus Huanglongbing）细菌，了解为奠定基础。分别称取样品5 g，用无菌水适量梯度稀释，各取200 μL稀释液涂布于不，重复3次。根据菌落形态、颜色、边缘状态、透明度、表面干湿状态等特征判断分离出的菌落种类并计数。按常规方法挑取单菌落纯化后保存，供测试鉴定。1.3.1 细菌脂肪酸的提取 ①细菌培养条件：TSBA平板培养基（30 g胰蛋白胨大豆肉汤（TSB，购于Fisher公司）+15 g琼脂+1 L水），四线划线法，培养温度（28±1） ℃，培养时间（24±2） h。②获菌：用接种环挑取3-5环（湿重约40 mg）菌落置入干净、干燥有螺旋盖的试管中（最佳获菌区域为第3区）。③皂化：加入（1.0±0.1） mL皂化试剂（45