First report of a 'Candidatus Phytoplasma phoenicium'-related strain (16SrIX-C) associated with phyllody disease of Reseda lutea

Matthiola incana R. Br. (Fam: Brassicaceae) is an ornamental, commonly known as hoary stock has an extremely fragrant flowers, which blooms in dense clusters in a large variety of colors. During a survey of flower nurseries in March 2019 at Indian Institute of Sugarcane Research campus, Lucknow, floral virescence (MiV) symptoms (Fig. 1 A, B) were observed in M. incana pots with an incidence of over 40%. Leaf yellows symptoms were also observed on a weed Acalypha indica (AiLY) in Matthiola nursery (Fig. 1 C). Nested PCR assays were carried out to detect and identify the possible association of phytoplasmas with MiV and AiLY symptoms. Three each of symptomatic MiV and AiLY samples and two non-symptomatic samples were collected and processed for DNA extraction from the leaf midrib by CTAB method. Hishimonus phycitis (HP) (Hemiptera: Cicadellidae) leafhopper feeding on MiV symptomatic plants was also collected and DNA was extracted. The DNA of 8 symptomatic and 4 non-symptomatic plants and from the 10 leafhopper was used as a template for PCR assays. Phytoplasma specific 16Sr RNA gene specific primers (P1/P7 and 3Far/3Rev; Schneider et al. 1995; Manimekalai et al. 2010) and multilocus genes’ specific primer pairs for secA (SecAfor1/SecArev3;SecAfo5r/SecARev2; Bekele et al. 2011), secY (SecYF1(VI)/SecYR1(VI);SecYF2(VI)/SecYR1(VI); Lee et al. 2010) and rp genes (rpFIC/rp(I)R1A; rp(VI)F2/ rp(VI)R2; Martini et al. 2007) were employed as previously described. Amplified products of ~1.3kb, ~600bp, ~1.7kb and ~1.0kb of 16S rRNA, secA, secY and rp genes of phytoplasma were consistently amplified in all the MiV and AiLY samples and in the HP leafhopper. No amplifications were achieved in any of the asymptomatic plant samples. Amplified products of all the four genes of MiV, AiLY and HP isolates were purified, sequenced and submitted in GenBank. Sequence comparison and phylogeny analysis of the sequences of the four genes of MiV, AiLY and HP isolates revealed 99% - 100% sequence identity and clustering with clover proliferation phytoplasma related strains (16SrVI group)(Fig.2 A,B,C and D). The virtual RFLP analysis of 17 restriction endonucleases corresponding to the 16S rDNA sequence of MiV, AiLY and HP phytoplasma strains by pDraw program, assigned them into a novel phytoplasma subgroup strain under 16SrVI group, since its HpaII restriction profile was different to earlier classified 16SrVI subgroups but was very close to16SrVI-E subgroup (GenBank acc. no. AY270156) (Fig 3). Earlier, peanut witches’ broom (16SrII-A) phytoplasma was identified associated with M. incana from Italy (Davino et al. 2007). However, the association of clover proliferation phytoplasma (16SrVI) related strain associated with virescence symptom of M. incana is the first report in world. The weed (A. indica) and HP leafhopper were also reported as additional hosts of 16SrVI subgroup related new strain in India, which needs further investigation. The report of a new host and new subgroup of clover proliferation phytoplasma related strain in India is having an epidemiological significance and warrants attention.

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First report of a ‘CandidatusPhytoplasma asteris’-related strain associated with a yellows disease of black pepper (Piper nigrum) in India

Plant Pathology ◽

10.1111/j.1365-3059.2009.02050.x ◽

2009 ◽

Vol 58 (4) ◽

pp. 789-789 ◽

Cited By ~ 8

Author(s):

C. R. Adkar-Purushothama ◽

P. Casati ◽

F. Quaglino ◽

G. Durante ◽

P. A. Bianco

Keyword(s):

Black Pepper ◽

Piper Nigrum ◽

Related Strain ◽

First Report

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First report of a 'Candidatus Phytoplasma asteris'-related strain (16SrI-B) associated with Morus alba (white mulberry) witches' broom in Iran

New Disease Reports ◽

10.5197/j.2044-0588.2020.041.025 ◽

2020 ◽

Vol 41 ◽

pp. 25

Author(s):

M. Salehi ◽

E. Salehi ◽

S.A. Esmaeilzadeh-Hosseini

Keyword(s):

Morus Alba ◽

Related Strain ◽

First Report ◽

White Mulberry

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First report of a 'Candidatus Phytoplasma australasia'-related strain (16SrII-D subgroup) associated with alfalfa leaf chlorosis disease in Turkey

New Disease Reports ◽

10.5197/j.2044-0588.2020.042.020 ◽

2020 ◽

Vol 42 ◽

pp. 20

Author(s):

H. Ayvaci ◽

M.E. Guldur ◽

E. Simsek ◽

M. Dikilitas

Keyword(s):

Related Strain ◽

First Report ◽

Leaf Chlorosis ◽

Alfalfa Leaf

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First report of a 'Candidatus Phytoplasma trifolii'-related strain (16SrVI-A) associated with white leaf disease in Spirea japonica

Australasian Plant Disease Notes ◽

10.1007/s13314-021-00441-z ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Eray Şimşek ◽

Hümeyra Ayvacı ◽

Havva Akkurak ◽

Murat Dikilitas ◽

Mehmet Ertuğrul Güldür

Keyword(s):

Related Strain ◽

First Report ◽

Leaf Disease ◽

White Leaf ◽

White Leaf Disease

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First Report of Candidatus Phytoplasma solani Associated with Potato Plants in Greece

Plant Disease ◽

10.1094/pdis-05-14-0488-pdn ◽

2014 ◽

Vol 98 (12) ◽

pp. 1739-1739 ◽

Cited By ~ 3

Author(s):

M. C. Holeva ◽

P. E. Glynos ◽

C. D. Karafla ◽

E. M. Koutsioumari ◽

K. B. Simoglou ◽

...

Keyword(s):

16S Rdna ◽

Nested Pcr ◽

Pathogenic Bacteria ◽

Related Strain ◽

Northern Greece ◽

First Report ◽

Potato Plants ◽

Pcr Product ◽

16S Rdna Pcr ◽

Stolbur Phytoplasma

In August 2013, potato plants (Solanum tuberosum) cv. Banba displaying symptoms resembling those caused by Candidatus Phytoplasma solani (potato stolbur phytoplasma) were observed in a 2-ha field in the area of the Peripheral Unit of Drama (northern Greece). The plants were 10 weeks old and their symptoms included reddening and upward rolling of leaflets, reduced size of leaves, shortened internodes, and aerial tuber formation. Incidence of affected plants was estimated to be 40% in the field. Four symptomatic potato plants were collected for laboratory testing of possible phytoplasma infection. From each of these four plants, total DNA was extracted from mid veins of reddish leaflets from apical shoot parts and of leaflets emerging from aerial tubers, using a phytoplasma enrichment procedure (1). A nested PCR using the phytoplasma universal 16S rRNA primer pairs: P1/P7 followed by R16F2n/R16R2 (3) amplified the expected ~1.2-kb 16S rDNA fragment in all four symptomatic potato plants. No amplification was observed with DNA similarly extracted from leaflets of asymptomatic potato plants of the same variety collected from an apparently healthy crop. One of the four 1.2-kb nested 16S rDNA PCR products was gel purified, cloned into the pGEM-T-easy plasmid vector (Promega, Madison, WI), and sequenced by Beckman Coulter Genomics (United Kingdom). At least twofold coverage per base position of the cloned PCR product was achieved. BLAST analysis showed that the obtained sequence of the PCR 16S rDNA product was: i) 100% identical to several GenBank sequences of Ca. P. solani strains, including strains detected previously in Greece infecting tomato (GenBank Accession No. JX311953) and Datura stramonium (HE598778 and HE598779), and ii) 99.7% similar to that of the Ca. P. solani reference strain STOL11 (AF248959). Furthermore, analysis by iPhyClassifier software showed that the virtual restriction fragment length polymorphism (RFLP) pattern of the sequenced PCR 16S rDNA product is identical (similarity coefficient 1.00) to the reference pattern of the 16SrXII-A subgroup (AF248959). The sequence of this PCR product was deposited in NCBI GenBank database under the accession no. KJ810575. The presence of the stolbur phytoplasma in all four symptomatic potato plants examined was further confirmed by nested PCR using the stolbur-specific STOL11 primers (3) targeting non-ribosomal DNA. Based on the observed symptoms in the field and laboratory molecular examinations, we concluded that the potato plants were infected by a Ca. P. solani related strain. The stolbur disease has been previously reported in Greece affecting tomato (2,5) and varieties of D. stramonium (4). To our knowledge, this is the first report of a Ca. P. solani related strain infecting a potato crop in Greece. As northern Greece is a center of potato production, the source of this pathogen is to be investigated. References: (1) U. Ahrens and E. Seemuller. Phytopathology 82:828, 1992. (2) A. S. Alivizatos. Pages 945-950 in: Proceedings of the 7th International Conference of Plant Pathogenic Bacteria. Academiai Kiado, Budapest, Hungary, 1989. (3) J. Jović et al. Bull. Insectol. 64:S83, 2011. (4) L. Lotos et al. J. Plant Pathol. 95:447, 2013. (5) E. Vellios and F. Lioliopoulou. Bull. Insectol. 60:157, 2007.

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