Severe little leaf and excessive shoot proliferation symptoms typical of the peanut little leaf (PnLL) disease were observed in different peanut (Arachis hypogaea) fields, variety K-6, at Kadiriand Gooty regions, of Andhra Pradesh, India, during the monsoon season of 2018–19. Disease incidence was 2–3%. The phytoplasma etiology of the recorded disease was confirmed by amplifying ~1.25 kb DNA products of 16S rRNA gene from symptomatic plants using nested polymerase chain reaction with phytoplasma specific primers, P1/P7 and R16F2n/R16R2, respectively. A further confirmation of the phytoplasma etiology was obtained by amplification of 840 bp, 1094 bp and 465 bp DNA fragments using phytoplasma specific primers targeting secA, tuf and SAP11 genes, respectively. Sequence and phylogenetic analyses of the amplified 16S rRNA, secA, tuf and SAP11 genes revealed that the detected phytoplasma is a member of the peanut witches’-broom phytoplasma group or 16SrII group (‘Candidatus Phytoplasma australasia’). Also, on the basis of computer-simulated RFLP (= in silico RFLP) analysis of amplified 16S rRNA gene, the detected phytoplasma was assigned to subgroup D (16SrII-D). Parthenium hysterophorus plants showing witches’ broom and Cleome viscosa plants with little leaf symptoms, both collected in the mentioned peanut fields were also infected by similar strain of phytoplasma which proved to be identical with each of the molecular marker employed to the peanut-infecting agent in India. This is the first report on the association of ‘Ca. P. australasia’ (16SrII-D subgroup phytoplasma) with PnLL disease in India.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Ahrens U, Seemüller E (1992) Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828–832
Al-Subhi AM, Hogenhout SA, Al-Yahyai RA, Al-Sadi AM (2018) Detection, identification, and molecular characterization of the 16SrII-D phytoplasmas infecting vegetable and field crops in Oman. Plant Dis 102:576–588
Anonymous (2018) Agricultural and Processed Food Products Exports Development Authority, Ministry of Commerce and Industry, Government of India
Arya SS, Salve AR, Chauhan S (2016) Peanuts as functional food: a review. J Food Sci Technol 53:31–41
Bertaccini A, Lee M (2018) Phytoplasmas: an update. In: Rao GP, Bertaccini A, Fiore N, Liefting LW (eds) Phytoplasmas: plant pathogenic Bacteria-I. Springer, Singapore, pp 1–29
Deng S, Hiruki C (1991) Amplification of 16S rRNA genes from culturable and nonculturable mollicutes. J Microbiol Methods 14:53–61
Gundersen DE, Lee I-M (1996) Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathol Medit 35:114–151
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucl Acids SympSer 4:95–98
Hodgetts J, Boonham N, Mumford R, Harrison N, Dickinson M (2008) Phytoplasma phylogenetics based on analysis of secA and 23S rRNA gene sequences for improved resolution of candidate species of ‘Candidatus Phytoplasma’. Int J Syst Evol Microbiol 58:1826–1837
Kumar S, Stecher G, Tamura K (2016) MEGA 7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874
Martini M, Delić D, Liefting L, Montano H (2018) Phytoplasmas infecting vegetable, Pulseand oil crops. In: Rao GP, Bertaccini A, Fiore N, Leiftwing LW (eds) Phytoplasmas: plant pathogenic Bacteria-I. Springer, Singapore, pp 31–65
Pande S, Rao JN (2000) Changing scenario of groundnut diseases in Andhra Pradesh, Karnataka, and Tamil Nadu states of India. International Arachis Newsletter 20:42–44
Rao GP, Tiwari AK, Kumar S, Baranwal VK (2014) Identification of sugarcane grassy shoot-associated phytoplasma and one of its putative vectors in India. Phytoparasitica 42:349–354
Rao GP, Madhupriya TV, Manimekalai R, Tiwari AK, Yadav A (2017) A century progress of research on phytoplasma diseases in India. Phytopathogenic Mollicutes 7:1–38
Thorat V, Bhale U, Sawant V, More V, Jadhav P, Mane SS, Nandanwar RS, Tripathi S, Yadav A (2016) Alternative weed hosts harbors 16SrII group phytoplasmas associated with little leaf and witches’ broom diseases of various crops in India. Phytopathogenic Mollicutes 6:50–55
Weintraub PG, Beanland L (2006) Insect vectors of phytoplasmas. Annu Rev Entomol 51:91–111
Zhao Y, Wei W, Lee M, Shao J, Suo X, Davis RE (2009) Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII). Int J Syst Evol Microbiol 59:2582
The authors are thankful to the DBT, New Delhi, India, for providing financial assistance under the DBT-Twinning Research Project. Also, the authors wish to express sincere thanks to the Head, Division of Plant Pathology, and the Director, Indian Agricultural Research Institute, for providing laboratory facilities.
Conflict of interest
The authors declare that they have no competing interests.
This article does not contain any studies with human participants or animals performed by any of the authors.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Mitra, S., Kumar, M., Vemana, K. et al. Multilocus sequence analysis of a ‘Candidatus Phytoplasma australasia’-related strain associated with peanut little leaf disease in India. J Plant Pathol 103, 311–316 (2021). https://doi.org/10.1007/s42161-020-00704-1
- Arachis hypogaea
- 16SrII-D subgroup
- Multilocus genes
- Oilseed crop