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Molecular characterization of phytoplasma strains associated with brinjal little leaf and screening of cultivated and wild relatives of eggplant cultivars for disease resistance

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Abstract

Brinjal little leaf (BLL) is one of the most important and widespread disease of eggplant associated with a phytoplasma in India. It severely infects eggplant cultivation in India and causes serious economic losses. Severe incidence (8 to 30%) of the disease was recorded in three districts of Uttar Pradesh (Varanasi, Mirzapur and Jaunpur) state of India during 2015 and 2016. A total of 58 symptomatic BLL leaf samples were collected from the surveyed fields and processed for nested PCR assays using phytoplasma-specific primer pairs (P1/P7, R16F2n/R16R2). Pair wise sequence identity and phylogeny analysis of 16S rRNA gene sequences of BLL isolates in the study confirmed association of Ca. P. trifoli (16SrVI group) with the BLL symptomatic samples. Association of similar strain of phytoplasma was identified in Hishimonus phycitis collected in brinjal fields at all the locations, utilizing a similar set of primers pairs as described above. The population of H. phycitis was positively correlated with the incidence of BLL disease in the fields. The phytoplasma indexing of 55 eggplants varieties and 17 wild Solanum species through PCR assays revealed that one eggplant cultivated variety (Uttara) and 17 wild Solanum species were found immune, one resistant (Pusa Ankur) and 12 eggplant varieties/lines were found moderately resistant to BLL. However, all the 17 wild Solanum species were recorded free from phytoplasmas in PCR assays. Further, biochemical analysis of the resistant eggplant varieties showed higher peroxidase and polyphenol oxidase enzymatic activities along with the increased total phenols content. These resistant varieties identified in the present study can be utilized as pre-breeding materials to breed and develop eggplant resistance to BLL phytoplasma disease.

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References

  • Ashwathappa, K. V., Venkataravanappa, V., Reddy, C. N. L., Swarnalatha, P., & Reddy, M. K. (2019). First report of ‘Candidatus Phytoplasma aurantifolia’(16SrII-A) associated with strawflower phyllody. Australasian Plant Disease Notes, 14, 28.

    Article  CAS  Google Scholar 

  • Ashwathappa, K. V., Reddy, C. N. L., Venkataravanappa, V., Suryanarayana, V. S., Gurivi Reddy, M., & Rao, G. P. (2020). A new strain of ‘Candidatus Phytoplasma asteris’ associated with bottle gourd phyllody disease in South India. Phytopathogenic Mollicutes, 10(1), 43–49.

    Article  Google Scholar 

  • Azadvar, M., & Baranwal, V. K. (2012). Multilocus sequence analysis of phytoplasma associated with brinjal little leaf disease and its detection in Hishimonas phycitis in India. Phytopathogenic Mollicutes, 2, 15–21.

    Article  Google Scholar 

  • Baker, C. J., & Orlandi, E. W. (1995). Active oxygen in plant pathogenesis. Annual Review Phytopathology, 33, 299–321.

    Article  CAS  Google Scholar 

  • Bendich, A. (1996). Antioxidants vitamins and immune response. Vitamins and Hormones, 52, 35–62.

    Article  CAS  PubMed  Google Scholar 

  • Bertaccini, A., Davis, R. E., & Lee, I.-M. (1992). In vitro micropropagation for maintenance of mycoplasmalike organisms in infected plant tissues. Horticultural Science, 27(9), 1041–1043.

    Google Scholar 

  • Bertaccini, A., & Duduk, B. (2009). Phytoplasma and phytoplasma diseases: a review of recent research. Phytopathology Mediterranean, 48, 355–378.

    CAS  Google Scholar 

  • Bingham, I.J., Newton, A.C. (2009). Crop tolerance of foliar pathogens: possible mechanisms and potential for exploitation, in Disease Control in Crops – Biological and Environmentally Friendly Approaches, ed Walters D. (Chichester, UK: Wiley-Blackwell ), 142–161.

  • Chakrabarti, A. K., & Choudhury, B. (1975). Breeding brinjal resistant to little leaf disease. Proceedings of the National Academy of Sciences, India, Section B: Biological Sciences, 41(4), 379–385.

    Google Scholar 

  • Das, A. K., Mitra, D. K., & Ahlawat, Y. S. (2000). Detection of eggplant little leaf phytoplasma in phloem elements isolated by enzyme treatment. Indian Phytopathology, 53(2), 146–148.

    CAS  Google Scholar 

  • Deng, S., & Hiruki, C. (1991). Genetic relatedness between two non-culturable mycoplasma like organisms revealed by nucleic acid hybridization and polymerase chain reaction. Phytopathology, 81, 1475–1479.

    Article  Google Scholar 

  • Devi, S. N., Mathew, S. K., & Vahab, M. A. (1995). Evaluation of brinjal for resistance to little leaf disease. South Indian Horticulture, 43, 120–121.

    Google Scholar 

  • Doyle, J. J., & Doyle, J. L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12, 13–15.

    Google Scholar 

  • Dutta, D. S., Kalita, M. K., Deb Nath, P., Rahman, S., & Boro, R. (2020). Incidence, symptomatology, molecular detection and yield loss assessment of brinjal little leaf disease in Assam, India. Phytopathogenic Mollicutes, 10(2), 203–213.

    Article  Google Scholar 

  • Galtier, N., Gouy, M., & Gautier, C. (1996). SEA VIEW and PHYLO WIN: two graphic tools for sequence alignment and molecular phylogeny. Computer Applications in the Biosciences, 12, 543–548.

    CAS  PubMed  Google Scholar 

  • Gundersen, D. E., Lee, I. M., Rehner, S. A., Davis, R. E., & Kingsbury, D. T. (1994). Phylogeny of mycoplasmalike organisms (phytoplasmas): a basis for their classification. Journal of Bacteriology, 176, 5244–5254.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta, M. K., Samad, A., Shasany, A. K., Ajayakumar, P. V., & Alam, M. (2009). First report of a 16SrVI ‘Candidatus Phytoplasma trifolii’ isolate infecting Norfolk Island pine (Araucaria heterophylla) in India. Plant Pathology, 59, 399.

    Article  Google Scholar 

  • Heinrich, M., Botti, S., & Caprara, L. (2001). Improved detection methods for fruit tree phytoplasmas. Plant Molecular Biology Reporter, 19, 169–179.

    Article  CAS  Google Scholar 

  • Hemmati, C., Al-Subhi, A. M., & Al-Housni, M. T. (2020). Molecular detection and characterization of a 16SrII-D phytoplasma associated with streak yellows of date palm in Oman. Australasian Plant Disease Notes, 15, 35.

    Article  CAS  Google Scholar 

  • 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’. International Journal of Systematic and Evolutionary Microbiology, 58, 1826–1837.

    Article  CAS  PubMed  Google Scholar 

  • Jabeen, N., Ahmed, N., Ghani, M. Y., & Sofi, P. A. (2009). Role of phenolic compounds in resistance to chilli wilt. Communications in Biometry and Crop Science, 4(2), 52–61.

    Google Scholar 

  • Khasa, E., Taloh, A., Gopala., Prabha, T., Rao G.P.(2016). Molecular characterization of phytoplasmas of ‘Clover proliferation’ group associated with three ornamental plant species in India. 3 Biotech, 6, 237.

  • Kumar, A., Mali, P. C., & Manga, V. K. (2010). Changes of some phenolic compounds and enzyme activities on infected pearl millet caused by Sclerospora graminicola. International Journal of Plant Physiology and Biochemistry, 2(1), 6–10.

    CAS  Google Scholar 

  • Kumar, M., Madhupriya, Rao, G.P. (2017). Molecular characterization, vector identification and sources of phytoplasmas associated with brinjal little leaf disease in India. 3 Biotech,7, 7

  • Kumar, M. (2015). Genetic Diversity and Natural Spread Sources of Brinjal Little Leaf Phytoplasma. M.Sc. Thesis. Div. Pl. Pathol., Indian Agricultural Research Institute, New Delhi, India. p.103.

  • Kumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870–1874.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee, I.-M., Gundersen, D. E., Hammond, R. W., & Davis, R. E. (1994). Use of mycoplasma like organism (MLO) group specific oligonucleotide primers from nested-PCR assays to detect mixed-MLO infections in a single host plant. Phytopathology, 84, 559–566.

    Article  CAS  Google Scholar 

  • Martini, M., Lee, I.-M., Bottner, K. D., Zhao, Y., Botti, S., Bertaccini, A., Harrison, N. A., Carraro, L., Marcone, C., Khan, A. J., & Osler, R. (2007). Ribosomal protein gene-based phylogeny for finer differentiation and classification of phytoplasmas. International Journal of Systematic and Evolutionary Microbiology, 57, 2037–2051.

    Article  CAS  PubMed  Google Scholar 

  • Matto, A., & Diamond, A. E. (1963). Symptoms of Fusarium wilt in relation to quality of fungus and enzyme activity in tomato stem. Phytopathology, 53, 574–575.

    Google Scholar 

  • Melo, G. A., Shimizu, M. M., & Mazzafera, P. (2006). Polyphenoloxidase activity in coffee leaves and its role in resistance against coffee leaf miner and coffee leaf rust. Phytochemistry, 67, 277–285.

    Article  CAS  PubMed  Google Scholar 

  • Memane, S., Joi, M., & Kale, P. (1987). Screening of chilli cultivars against leaf curl complex. Current Research Reporter, Mahatma Phule Agricultural University., 3, 98–99.

    Google Scholar 

  • Mitra, D.K. (1993). Little leaf, a serious disease of eggplant (Solanum melongena). In: Raychaudhuri, S.P., Teakle, D.S. (Eds.), Management of plant diseases caused by fastidious prokaryotes. Associated Publishing Co, , pp. 73–78.

  • Namba, S. H., Oyaizu, H., Kato, S., Iwanami, S., & Tsuchizaki, T. (1993). Phylogenetic diversity of phytopathogenic mycoplasma-like organisms. International Journal of Systematic Bacteriology, 43, 461–467.

    Article  CAS  PubMed  Google Scholar 

  • Namrata, J., Singh, M., Dubey, R.S., Venkataramanappa, V., Datta D. (2013). Phytochemicals and antioxidative enzymes defence mechanism on occurrence of yellow vein mosaic disease of pumpkin (Cucurbita moschata). 3 Biotech, 3, 287–295.

  • Rao, G.P., Bertaccini A., Fiore N., Liefting Lia W. (2018). Editors Phytoplasmas: Plant Pathogenic Bacteria – I, Characterization and Epidemiology of Phytoplasma- Associated Diseases SpringerNature Singapore Pte Ltd. , pp. 345.(ISBN 978-981-13-0118-6).

  • Rao, G. P., & Kumar, M. (2017). World status of phytoplasma diseases associated with eggplant. Crop Protection, 96, 22–29.

    Article  Google Scholar 

  • Rao, G. P. (2021). Our understanding about phytoplasma research scenario in India. Indian Phytopathology, 74, 371–401.

    Article  Google Scholar 

  • Salehi, M., Izadpanah, K., Siampour, M., & Bagheri, A. (2007). Transmission of ‘Candidatus Phytoplasma aurantifolia’ to Bakraee (Citrus reticulata hybrid) by Feral Hishimonas phycitis leafhoppers in Iran. Plant Disease, 91, 466.

    Article  CAS  PubMed  Google Scholar 

  • Samad, A., Ajayakumar, P. V., Shasany, A. K., Gupta, M. K., Alam, M., & Rastogi, S. (2008). Occurrence of a clover proliferation (16SrVI) group phytoplasma associated with little leaf disease of Portulaca grandiflora in India. Plant Disease, 92, 832.

    Article  CAS  PubMed  Google Scholar 

  • Samad, A., Shasany, A. K., Gupta, S., Ajayakumar, P. V., Darokar, M. P., & Khanuja, S. P. S. (2006). First report of a 16SrVI group phytoplasma associated with witches’ broom disease on Withania somnifera. Plant Disease, 90, 248.

    Article  CAS  PubMed  Google Scholar 

  • Saranya, S. S., & Umamaheswaran, K. (2014). Symptomatology, transmission and molecular detection of phytoplasma infecting brinjal (Solanum melongena L.). International Journal of Applied and Pure Science and Agriculture, 1, 2394–2332.

    Google Scholar 

  • Sertkaya, G., Martini, M., Musetti, R., & Osler, R. (2007). Detection and molecular characterization of phytoplasmas infecting sesame and solanaceous crops in Turkey. Bulletin of Insectology, 60, 141–142.

    Google Scholar 

  • Shannon, L., Kay, E., & Lew, J. (1996). Peroxidase Isozymes from Horseradish Roots. I. Isolation and Physical Properties. Journal of Biological Chemistry, 241, 2166–1966.

    Google Scholar 

  • Shivalingaiah, & Umesha, S. (2016). Study of involvement of peroxidase and polyphenol oxidase in imparting resistance to bacterial blight disease in Oryza sativa varieties. European Journal of Biotechnology and Biosciences, 4(7), 5–10.

    Google Scholar 

  • Siampour, M., Izadpanah, K., Afsharifar, A. R., Salehi, M., & Taghizadeh, M. (2006). Detection of phytoplasma in insects collected in witches’ broom affected lime groves. Iranian Journal of Plant Pathology, 42, 139–158.

    Google Scholar 

  • Singh, N., Rao, G. P., Madhupriya, U., & P.P. (2012). ‘Candidatus Phytoplasma trifolii’ associated with little leaf and witches’ broom disease of Datura stramonium L. in India. Phytopathogenic Mollicutes, 2, 69–71.

    Article  CAS  Google Scholar 

  • Smart, C. D., Schneider, B., Blomquist, C. L., Guerra, L. J., Harrison, N. A., & Ahrens, U. (1996). Phytoplasma specific PCR primers based on sequences of 16S-23SrRNA spacer region. Applied and Environmental Microbiology, 62, 2988–2993.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thomas, K.M., Krishnaswami, C.S. (1939). ‘Little leaf’, a transmissible disease of brinjal. Proceedings of the National Academy of Sciences, 10, 201–212.

  • Venkataravanappa, V., Ashwathappa, K. V., Hemachandra Reddy, P., Reddy, C. N. L., Jalali, S., & Reddy, M. K. (2018b). ‘Candidatus Phytoplasma’ belonging to the 16SrVI phytoplasma group, is associated with witches broom disease of Azadirachta indica in India. Australasian Plant Disease Notes, 13, 28.

    Article  Google Scholar 

  • Venkataravanappa, V., Prasanna, H. C., Reddy, C. N. L., & Reddy, M. K. (2018a). Molecular detection and characterization of phytoplasma in association with begomovirus in eggplant. Acta virologica, 62, 246–258.

    Article  CAS  PubMed  Google Scholar 

  • Venkataravanappa, V., Reddy, C. N. L., Swarnalatha, P., Shankarappa, K. S., & Reddy, M. K. (2017). Detection and characterization of ‘Candidatus Phytoplasma asteris’ associated with little leaf disease of bitter gourd from India by 16S rRNA phylogenetic and RFLP (in vitro and virtual) analysis. Archives of Biological Sciences, 69(4), 707–714.

    Article  Google Scholar 

  • Wei, W., Davis, R. E., Lee, I.-M., & Zhao, Y. (2007). Computer-simulated RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. International Journal of Systematic and Evolutionary Microbiology, 57, 1855–1867.

    Article  CAS  PubMed  Google Scholar 

  • Wei, W., Lee, I.-M., Davis, R. E., Suo, X., & Zhao, Y. (2008). Automated RFLP pattern comparison and similarity coefficient calculation for rapid delineation of new and distinct phytoplasma 16Sr subgroup lineages. International Journal of Systematic and Evolutionary Microbiology, 58, 2368–2377.

    Article  CAS  PubMed  Google Scholar 

  • Zaim, M., & Samad, A. (1995). Association of phytoplasmas with a witches’ broom disease of Withania somnifera (L.) Dunal in India. Plant Science, 109, 225–229.

    Article  CAS  Google Scholar 

  • Zhao, Y., Wei, W., Lee, I.-M., Shao, J., Suo, X., & Davis, R. E. (2009). Construction of an interactive online phytoplasma classification tool, iPhyClassifier, and its application in analysis of the peach X-disease phytoplasma group (16SrIII). International Journal of Systematic and Evolutionary Microbiology, 59, 2582–2593.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The work was carried out under the grants from the Indian Council of Agricultural Research (ICAR), New Delhi, India in the form of ICAR-Outreach Programme on Management of Sucking Pests in Horticultural Crops (ORP-SP). The authors gratefully acknowledge the help and support given by the Director and Head, Division of Crop Protection, ICAR-Indian Institute of Vegetable Research, Varanasi, India to carry out the experiments.

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Authors and Affiliations

  1. Central Horticultural Experimental Station, ICAR-IIHR, Chettalli-571248, Madikeri, Karnataka, India

    V. Venkataravanappa

  2. ICAR- Indian Institute of Pulses Research, Regional Research Center, UAS Campus, Dharwad, Karnataka, 580005, India

    M.H. Kodandaram

  3. Division of Crop Sciences, ICAR-Central Research Institute for Dryland Agriculture, Santhosh Nagar, Hyderabad, Telangana, 500059, India

    M. Manjunath & B. Sarkar

  4. ICAR-Indian Institute of Vegetable Research, Varanasi, Uttar Pradesh, 221305, India

    V. Venkataravanappa, M.H. Kodandaram, M. Manjunath, Neha S. Chauhan, K. Nagendran & Shailesh K. Tiwari

  5. Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110012, India

    Govind Pratap Rao

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  1. V. Venkataravanappa
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Venkataravanappa, V., Kodandaram, M., Manjunath, M. et al. Molecular characterization of phytoplasma strains associated with brinjal little leaf and screening of cultivated and wild relatives of eggplant cultivars for disease resistance. Eur J Plant Pathol 162, 433–453 (2022). https://doi.org/10.1007/s10658-021-02414-5

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