Molecular characterization of phytoplasmas of ‘Clover proliferation’ group associated with three ornamental plant species in India

Khasa, Ekta; Gopala,; Taloh, Aido; Prabha, T. N.; Madhupriya,; Rao, G. P.

doi:10.1007/s13205-016-0558-8

Cited by 21 publications

(11 citation statements)

References 17 publications

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“…Araucaria little leaf (Gupta et al 2009), brinjal little leaf (Azadvar and Baranwal 2012), Withania little leaf (Zaim and Samad 1995;Samad et al 2006), Portulaca little leaf (Samad et al 2008), Datura little leaf (Raj et al 2009;Singh et al 2012) and Calotropis gigantea leaf yellows (Madhupriya et al 2010). However, occurrence of 16SrVI-D subgroup phytoplasma was reported only on anantmool, lemongrass, sesame, Hibiscus rosa-sinensis, Saponaria officinalis and Allamanda cathartica (Madhupriya et al 2014(Madhupriya et al , 2015Khasa et al 2016). Hence, in our study, brinjal is reported to be an additional host of 'Ca.…”

Section: Discussionmentioning

confidence: 54%

Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India

2017

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Brinjal little leaf (BLL) is a widespread disease of phytoplasma etiology in India that induces severe economic losses. Surveys were conducted in eight brinjal-growing states of India during July 2014 to September 2015 and eighteen BLL samples showing little leaf, phyllody and witches' broom symptoms were collected for phytoplasma identification. Presence of phytoplasmas was confirmed in all the eighteen BLL samples using polymerase chain reaction with phytoplasma-specific primer pairs (P1/P6, R16F2n/R16R2). Pair wise sequence comparison and phylogenetic relationship of 16S rRNA gene sequences of BLL phytoplasma strains confirmed that sixteen out of eighteen BLL strains belonged to clover proliferation phytoplasma (16SrVI) group and two BLL strains (GKP-A and GKP-B) from Gorakhpur, Uttar Pradesh, were classified under 16SrII group. Further virtual RFLP analysis of 16S rDNA sequences allowed finer classification of BLL strains into 16SrII-D and 16SrVI-D subgroups. BLL phytoplasma strains belonging to 16SrVI-D subgroup were found as the most widespread phytoplasma strains associated with BLL disease in India. 16SrVI-D subgroup phytoplasma association with two symptomatic weed species viz. Cannabis sativa subsp. sativa at Noida, Uttar Pradesh and Portulaca oleracea at IARI fields, New Delhi was also confirmed by nested PCR assays with similar set of phytoplasma-specific primers, pairwise 16S rDNA sequence comparison, phylogeny and virtual RFLP analysis. Out of five identified leafhopper species from BLL-infected fields at Noida, Uttar Pradesh and Delhi, only Hishimonas phycitis was identified as carrier and natural vector of 16SrVI-D subgroup of phytoplasmas by nested PCR assays, sequence comparison, phylogeny, virtual RFLP analysis and transmission assays.

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Section: Discussionmentioning

confidence: 54%

Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India

2017

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“…The 16SrVI-D phytoplasma subgroup, have already been reported earlier as a major group of phytoplasma infecting several crops viz. brinjal (Rao and Kumar 2017), ornamental plants (Khasa et al 2016), medicinal plants (Madhupriya et al 2015) and sesame (Madhupriya et al 2015) in India.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of ‘Clover proliferation’ phytoplasma subgroup-D (16SrVI-D) associated with vegetables crops in India

Gopala

Khasa

Rao

et al. 2018

Physiol Mol Biol Plants

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Nine vegetable plants species exhibiting phytoplasma suspected symptoms of white/purple leaf, little leaf, flat stem, witches' broom, phyllody and leaf yellowing were observed in experimental fields at Indian Agricultural Research Institute, New Delhi from December 2015 to July 2016. Total DNA extracted from the three healthy and three symptomatic leaves of all the nine vegetables were subjected to PCR assays using phytoplasma specific primers P1/P7 followed by R16F2n/R16R2 and 3Far/3Rev to amplify the 16S rDNA fragments. No amplifications of DNA were observed in first round PCR assays with primer pair P1/P7 from any of the symptomatic samples. However, phytoplasma DNA specific fragments of ~ 1.3 kb were amplified from L. (two isolates), vr L. (one isolate) and L. (one isolate) by using 3Far/3Rev primer pair and 1.2 kb fragment was amplified from L. (one isolate) by using R16F2n/R16R2 primer pair. No DNA amplification was seen in other symptomatic vegetable samples of tomato, carrot, cucurbit, bitter gourd and species utilizing either P1/P7 primer pair followed by 3Far/3Rev or R16F2n/R16R2 primer pairs. Out of three leafhopper species collected from the symptomatic vegetable fields, only was found positive for association of phytoplasma. No DNA amplifications were observed in healthy plant samples and insects collected from non-symptomatic fields. Comparative sequence comparison analyses of 16S rDNA of positive found vegetable phytoplasma strains revealed 100% sequence identities among each other and with phytoplasma strains of 'clover proliferation' (16SrVI) group. Phytoplasma sequences, virtual RFLPs and phylogenetic analyses of 16S rDNA sequence comparison confirmed the identification of 16SrVI subgroup D strain of phytoplasmas in four vegetables and one leafhopper (HP) species. Further virtual RFLP analysis of 16S rDNA sequence of the vegetables phytoplasma strains confirmed their taxonomic classification with strains of 'clover proliferation' subgroup D. Since, feeding on symptomatic vegetable species in the study was also tested positive for the 16SrVI phytoplasma subgroup-D as of vegetables; it may act as potent natural reservoir of 16SrVI-D subgroup of phytoplasmas infecting vegetable and other important agricultural crops.

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“…In cucumber, phytoplasmas have been reported to be associated to disease, belonging to different groups (Montano et al 2006;Montano et al 2007). The phytoplasmas of 16SrVI clover proliferation (subgroup VI-A) have been reported to infect many plant species in the world (Khasa et al 2016;Zibadoost et al 2016;Choueiri et al 2007). However, there is no previous report of subgroup 16SrVI-A isolates infecting cucumber in Turkey.…”

Section: Discussionmentioning

confidence: 99%

Hıyar (Cucumis sativus L.) Bitkisinde Aynı Simptomatolojiye Sahip İki Farklı Fitoplazma etmeninin 16S rDNA'sının RFLP ve Nükleotid Dizi Analizleri ile Teşhisi ve Karakterizasyonu

Usta

Güller

Sipahioğlu

2017

Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi

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Phytoplasma-like symptoms were observed in cucumbers (Cucumis sativus L.) in Van province of Turkey. The major symptoms observed were severe dwarfing, witches' broom, rosetting, little leaf, and sterility of plants. Genomic DNA of 8 symptomatic and non-symptomatic plant leaves was isolated for the detection of pathogenic DNA. Of the 8 cucumber leaf samples tested by nested polymerase chain reaction (Nested-PCR), the four yielded the expected 1.25-kb DNA fragments when using universal primer pairs R16mF2/R16mR1 and R16F2n/R16R2. Randomly selected two DNA bands were further cloned into a proper plasmid vector. The recombinant plasmid DNA was sequenced bidirectionally. BLAST and virtual restriction fragment length polymorphism (RFLP) analyses of the 16S rDNA sequence revealed the presence of the "Candidatus Phytoplasma solani" (similarity coefficient 1.00) (GenBank accession no: KX977570) in one of the severely symptomatic cucumber samples and the "Candidatus Phytoplasma trifolii" (similarity coefficient 0.98) (GenBank accession no.: KR080212) in the other. The isolates were designated as Van-trifolii and Van-solani isolates, respectively. No significant differences were observed between the two different phytoplasmas' symptomatology. To the authors' knowledge, this is the first report of 'Ca. P. trifolii' and 'Ca. P. solani' in cucumbers in Turkey.

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Molecular characterization of phytoplasmas of ‘Clover proliferation’ group associated with three ornamental plant species in India

Cited by 21 publications

References 17 publications

Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India

Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India

Molecular characterization of ‘Clover proliferation’ phytoplasma subgroup-D (16SrVI-D) associated with vegetables crops in India

Hıyar (Cucumis sativus L.) Bitkisinde Aynı Simptomatolojiye Sahip İki Farklı Fitoplazma etmeninin 16S rDNA'sının RFLP ve Nükleotid Dizi Analizleri ile Teşhisi ve Karakterizasyonu

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