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运用PCR技术以柑橘黄龙病病原亚洲种、非洲种和美洲种特异性引物对赣南脐橙上7个代表性柑橘黄龙病样品进行扩增检测,结合限制性内切酶XbaⅠ对赣南脐橙上的黄龙病病原16S rDNA基因RFLP分析和测定的16SrDNA基因序列同GenBank登录的亚洲种、非洲种和美洲种的相应基因序列进行比对分析。结果表明,在赣南脐橙上检测到黄龙病病原亚洲种,未检测到非洲种和美洲种;赣南脐橙上黄龙病病原16S rDNA基因XbaⅠ酶切具有亚洲种图谱;16S rDNA基因序列比对分析表明7个赣南脐橙上的黄龙病病原间同源性为99.8%~100%,与亚洲种具有98.4%~99.9%的序列同源性,高于与非洲种/美洲种的序列同源性;与亚洲种间遗传距离小于与非洲种/美洲种间的遗传距离;系统聚类分析表明7个赣南脐橙上的黄龙病病原同亚洲种聚类在同一组群,具有较高的同源性和较近的亲缘关系,而与非洲种和美洲种在系统发育树中分属不同的组群,试验结果表明赣南脐橙上感染的黄龙病病原为亚洲种。
PCR-based PCR was used to detect the seven representative Citrus Huanglongbing samples in Gannan navel oranges by using the Asian, African and American species specific primers of citrus Huanglongbing pathogen, and the combination of restriction endonuclease XbaⅠ on Gannan navel orange The RFLP analysis and 16S rDNA gene sequence analysis of 16S rDNA gene of Huanglongbing pathogen were compared with the corresponding gene sequences of Asian, African and American species registered by GenBank. The results showed that the Asian species of Huanglongbing were detected in Gannan navel oranges and no African and American species were detected. The 16S rDNA gene Xba Ⅰ digested with the Asian species in Gannan navel orange, and the 16S rDNA gene sequence alignment analysis The results showed that there were 99.8% -100% of the pathogenicity among the four Gannan navel oranges, and 98.4% -99.9% homology with Asian species, which was higher than that of the African / American species ; The genetic distance to Asia was less than the genetic distance to African / American species. The phylogenetic analysis showed that the pathogenicity of yellow dragon’s disease on seven Gannan navel oranges was clustered in the same group with Asian species, with high homology Sexual and close relatives, but with African and American species in the phylogenetic tree belong to different groups, the experimental results show that Gannan navel orange yellow dragonfly pathogen infection of Asian species.