A novel 'Candidatus Phytoplasma asteris' subgroup 16SrI-(E/AI)AI associated with blueberry stunt disease in eastern Canada.

Phytoplasmas ('Candidatus Phytoplasma' species) are phytopathogenic bacteria vectored by insects and are associated with crop diseases that cause severe yield losses by affecting reproductive tissue development. Infection of northern highbush blueberry plants (Vaccinium corymbosum; Ericaceae) with phytoplasma leads to yield losses by altering plant development resulting in stunting and subsequent plant death. Samples collected from symptomatic blueberry plants in two important blueberry-producing areas in Canada, in the provinces of Québec and Nova Scotia, were analysed for the presence of DNA sequences associated with phytoplasma. Analysis of the 16S rRNA gene sequences demonstrated that the plants were infected with a strain of 'Candidatus Phytoplasma asteris', which was previously identified as blueberry stunt phytoplasma (BBS; 16SrI-E). Examination of further bacterial sequences revealed that two distinct 16S rRNA-encoding gene sequences were present in each sample in combination with a single chaperonin-60 (cpn60) sequence and a single rpoperon sequence, suggesting that this strain displays 16S rRNA-encoding gene sequence heterogeneity. Two distinct rrnoperons, rrnE and the newly described rrnAI, were identified in samples analysed from all geographic locations. We propose, based on the sequences obtained, delineating the new subgroup 16SrI-(E/AI)AI, following the nomenclature proposed for heterogeneous subgroups. To our knowledge, this is the first report of a heterogeneous phytoplasma strain affecting blueberry plants and associated with blueberry stunt disease.

[1]  Wei Wei,et al.  'Candidatus Phytoplasma wodyetiae', a new taxon associated with yellow decline disease of foxtail palm (Wodyetia bifurcata) in Malaysia. , 2017, International journal of systematic and evolutionary microbiology.

[2]  T. Dumonceaux,et al.  Molecular diagnostic assays based on cpn60 UT sequences reveal the geographic distribution of subgroup 16SrXIII-(A/I)I phytoplasma in Mexico , 2017, Scientific Reports.

[3]  T. Dumonceaux,et al.  Phytoplasma classification and phylogeny based on in silico and in vitro RFLP analysis of cpn60 universal target sequences , 2016, International journal of systematic and evolutionary microbiology.

[4]  T. Dumonceaux,et al.  Detection and identification of the heterogeneous novel subgroup 16SrXIII-(A/I)I phytoplasma associated with strawberry green petal disease and Mexican periwinkle virescence. , 2016, International journal of systematic and evolutionary microbiology.

[5]  F. Jan,et al.  The phytoplasma associated with purple woodnettle witches'‐broom disease in Taiwan represents a new subgroup of the aster yellows phytoplasma group , 2016 .

[6]  T. Dumonceaux,et al.  Molecular Diagnostic Tools for Detection and Differentiation of Phytoplasmas Based on Chaperonin-60 Reveal Differences in Host Plant Infection Patterns , 2014, PloS one.

[7]  T. Lowery,et al.  Occurrence of phytoplasmas in leafhoppers and cultivated grapevines in Canada , 2014 .

[8]  S. Namba,et al.  Exploring the phytoplasmas, plant pathogenic bacteria , 2014, Journal of General Plant Pathology.

[9]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[10]  M. Starović,et al.  First Report of Blueberry Reddening Disease in Serbia Associated with 16SrXII-A (Stolbur) Phytoplasma. , 2013, Plant Disease.

[11]  R. Davis,et al.  Characterization and molecular differentiation of 16SrI-E and 16SrIX-E phytoplasmas associated with blueberry stunt disease in New Jersey. , 2013, Molecular and cellular probes.

[12]  Yan Zhao,et al.  'Candidatus Phytoplasma pruni', a novel taxon associated with X-disease of stone fruits, Prunus spp.: multilocus characterization based on 16S rRNA, secY, and ribosomal protein genes. , 2013, International journal of systematic and evolutionary microbiology.

[13]  L. Conci,et al.  ESTABLISHMENT OF THE NEW SUBGROUP 16SrI-S (rr-rp) tuf-H BELONGING TO ‘Ca. PHYTOPLASMA ASTERIS’ IN WILD AND CULTIVATED PLANTS IN ARGENTINA , 2011 .

[14]  A. Bertaccini,et al.  The groEL gene as an additional marker for finer differentiation of ‘Candidatus Phytoplasma asteris'‐related strains , 2011 .

[15]  James K. Bonfield,et al.  Genome analysis Advance Access publication May 30, 2010 Gap5—editing , 2010 .

[16]  Wei Wei,et al.  Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII) , 2009, International journal of systematic and evolutionary microbiology.

[17]  Wei Wei,et al.  Automated RFLP pattern comparison and similarity coefficient calculation for rapid delineation of new and distinct phytoplasma 16Sr subgroup lineages. , 2008, International journal of systematic and evolutionary microbiology.

[18]  Y. Zhao,et al.  Ribosomal protein gene-based phylogeny for finer differentiation and classification of phytoplasmas. , 2007, International journal of systematic and evolutionary microbiology.

[19]  Wei Wei,et al.  Computer-simulated RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. , 2007, International journal of systematic and evolutionary microbiology.

[20]  D. Hegedus,et al.  First Report of "Candidatus Phytoplasma asteris"-Related Strains in Brassica rapa in Saskatchewan, Canada. , 2006 .

[21]  L. Camargo,et al.  Sequence heterogeneity in the 16S rDNA of tomato big bud phytoplasma belonging to group 16SrIII , 2006 .

[22]  J. Maas,et al.  POSSIBLE CAUSE OF EUROPEAN BLUEBERRY DISEASE IS RELATED TO NORTH AMERICAN MILKWEED YELLOWS PHYTOPLASMA , 2004 .

[23]  R. Davis,et al.  'Candidatus Phytoplasma asteris', a novel phytoplasma taxon associated with aster yellows and related diseases. , 2004, International journal of systematic and evolutionary microbiology.

[24]  D. Valiunas,et al.  New Group 16SrIII Phytoplasma Lineages in Lithuania Exhibit rRNA Interoperon Sequence Heterogeneity , 2002, European Journal of Plant Pathology.

[25]  R. Davis,et al.  Classification of aster yellows-group phytoplasmas based on combined analyses of rRNA and tuf gene sequences. , 2000, International journal of systematic and evolutionary microbiology.

[26]  D. Gundersen-Rindal,et al.  Revised Classification Scheme of Phytoplasmas based on RFLP Analyses of 16S rRNA and Ribosomal Protein Gene Sequences , 1998 .

[27]  Ing-Ming Lee,et al.  Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. , 1996 .

[28]  R. Beever,et al.  Sequence heterogeneity in the two 16S rRNA genes of Phormium yellow leaf phytoplasma , 1996, Applied and environmental microbiology.

[29]  C. Smart,et al.  Phytoplasma-specific PCR primers based on sequences of the 16S-23S rRNA spacer region , 1996, Applied and environmental microbiology.

[30]  D. Ramsdell,et al.  Populations of the sharpnosed leafhopper vector, Scaphytopius spp. and blueberry stunt infection periods in highbush blueberry , 1993 .

[31]  B. Sears,et al.  Evolutionary relationships of a plant-pathogenic mycoplasmalike organism and Acholeplasma laidlawii deduced from two ribosomal protein gene sequences , 1992, Journal of bacteriology.

[32]  T. Shigaki,et al.  'Candidatus Phytoplasma noviguineense', a novel taxon associated with Bogia coconut syndrome and banana wilt disease on the island of New Guinea. , 2018, International journal of systematic and evolutionary microbiology.

[33]  J. Scott,et al.  Identification and molecular characterization of the blueberry stunt phytoplasma in Canada , 2015 .

[34]  Wei Wei,et al.  Reductive evolution suggested from the complete genome sequence of a plant-pathogenic phytoplasma , 2004, Nature Genetics.

[35]  A. Kalvelytė,et al.  Differential amplification of sequence heterogeneous ribosomal RNA genes and classification of the 'Fragaria multicipita' phytoplasma. , 2003, Microbiological research.

[36]  R. Davis,et al.  Phytoplasma: phytopathogenic mollicutes. , 2000, Annual review of microbiology.

[37]  R. Hammond,et al.  Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms , 1993 .