To confirm phytoplasma infection,samples of pomegranate(Punica granatum L.)plants showing symptoms of fasciation were collected from an orchard located in Tai’an,Shandong Province,China.A fragment of approximately 1....To confirm phytoplasma infection,samples of pomegranate(Punica granatum L.)plants showing symptoms of fasciation were collected from an orchard located in Tai’an,Shandong Province,China.A fragment of approximately 1.2 kb was amplified with universal primers targeting the phytoplasma 16S r RNA gene from symptomatic pomegranate plants,while no fragment was obtained from healthy plants.The phytoplasma associated with the disease was designated as pomegranate fasciation(Po F).Two representative phytoplasma 16S r DNA gene sequences(Po F-Ch01 and Po F-Ch02)had 100%nucleotide sequence identity.The 16S r DNA sequence of Po F-Ch01 and Po F-Ch02 showed the highest similarity(99.6%)to that of‘P.granatum’phytoplasma isolate AY-PG,which belong to 16Sr I-B.Further phylogenetic analysis showed that Po F-Ch01 and Po F-Ch02 belonged to a cluster of 16Sr I subgroup members.In silico RFLP analysis indicated that Po F-Ch01 shared the highest similarity coefficient of 0.97 with reference strains of 16Sr I-B,M and N.Actual RFLP analysis of both enzymes Bst U I and Bfa I confirmed that of the virtual RFLP analysis.Combining these results,we concluded that Po F was a member of the‘Candidatus Phytoplasma asteris’group(16Sr I),and has very close relationship with 16Sr I-B subgroup.展开更多
基金supported by National Natural Science Foundation of China (Grant No. 31401718 )the Foundation for Excellent Young Scientists of Shandong Province (Grant No. BS2013NY012 )+1 种基金Specialized Research Fund for the Doctoral Program of Higher Education (Grant No. SRFDP, 20123702110013 BR0801 )
文摘To confirm phytoplasma infection,samples of pomegranate(Punica granatum L.)plants showing symptoms of fasciation were collected from an orchard located in Tai’an,Shandong Province,China.A fragment of approximately 1.2 kb was amplified with universal primers targeting the phytoplasma 16S r RNA gene from symptomatic pomegranate plants,while no fragment was obtained from healthy plants.The phytoplasma associated with the disease was designated as pomegranate fasciation(Po F).Two representative phytoplasma 16S r DNA gene sequences(Po F-Ch01 and Po F-Ch02)had 100%nucleotide sequence identity.The 16S r DNA sequence of Po F-Ch01 and Po F-Ch02 showed the highest similarity(99.6%)to that of‘P.granatum’phytoplasma isolate AY-PG,which belong to 16Sr I-B.Further phylogenetic analysis showed that Po F-Ch01 and Po F-Ch02 belonged to a cluster of 16Sr I subgroup members.In silico RFLP analysis indicated that Po F-Ch01 shared the highest similarity coefficient of 0.97 with reference strains of 16Sr I-B,M and N.Actual RFLP analysis of both enzymes Bst U I and Bfa I confirmed that of the virtual RFLP analysis.Combining these results,we concluded that Po F was a member of the‘Candidatus Phytoplasma asteris’group(16Sr I),and has very close relationship with 16Sr I-B subgroup.
