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Associated bacterial diversity of insecticide-susceptible and -resistant brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae) analyzed by culture-dependent and -independent methods

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Abstract

Nilaparvata lugens (Stål) is one of the major pests of rice throughout tropical and temperate Asia. Indiscriminate use of insecticides for suppressing N. lugens has resulted in the development of resistance to multiple insecticide classes, causing frequent control failures in the field. Analysis of gut bacterial diversity within an insect host is the initial step towards understanding the ecological roles of the symbionts. Present study aimed to survey the bacterial diversity associated with laboratory-reared (insecticide-susceptible) and field-collected (insecticide-resistant) populations of N. lugens by culture-dependent and PCR-Denaturing Gradient Gel Electrophoresis (DGGE) methods. Seventeen bacterial isolates were obtained by the culture-dependent method. Molecular characterization using the 16S rRNA gene and phylogenetic analysis revealed that the isolates belonged to Firmicutes and Proteobacteria. Taxonomic assignment placed these isolates into seven families representing 10 genera. Enterobacteriaceae was the most dominant family with its occurrence in four out of the five populations studied. The DGGE profiles indicated a low complex gut bacteria associated with N. lugens with limited number of bands. The Shannon-Wiener index ranged from 0.898 in insecticide-susceptible population to 0.946–1.035 in resistant populations. Sequencing and phylogenetic analysis revealed that the DGGE bands belonged to Firmicutes, Proteobacteria and Bacteriodetes. Results of this study illustrated that gut bacterial community associated with N. lugens is dominated by Proteobacteria and Firmicutes. Present findings could provide the basis for future work on the possible role of the bacterial symbionts in insecticide resistance and to formulate potential resistance management strategies.

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Acknowledgements

We acknowledge late Prof. D. K. Sidde Gowda, for his assistance and advice in the rearing of Nilaparvata lugens. Authors thank M/s Applied Maths, Germany, for providing evaluation license of Bionumerics software. The first author gratefully acknowledges the Department of Science and Technology (DST), Govt. of India, for the award of INSPIRE fellowship for Ph.D. work. We are thankful to staff of Regional Agricultural Research Station, Warangal and Nellore and Krishi Vigyan Kendra, Tiruchirappalli, for their kind help extended during field collection of Nilaparvata lugens. The Director, ICAR-NBAIR and the Director, ICAR-NIANP, Bengaluru, India, are thanked for providing necessary laboratory facilities for conducting this research.

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Authors and Affiliations

  1. Division of Molecular Entomology, Indian Council of Agricultural Research-National Bureau of Agricultural Insect Resources, Post Bag No. 2491, Hebbal, Bengaluru, Karnataka, 560 024, India

    V. M. Malathi, Sushil Kumar Jalali, R. Gandhi Gracy, Ravi Prabhakar More & Thiruvengadam Venkatesan

  2. Gut Microbiology Laboratory, Indian Council of Agricultural Research -National Institute of Animal Nutrition and Physiology, Adugodi, Bengaluru, Karnataka, 560 030, India

    Vazhakkala Jose Lyju & Apoothy Thulasi

  3. Department of Agricultural Microbiology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Madurai, -625104, India

    Rangasamy Anandham

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  1. V. M. Malathi
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Correspondence to Sushil Kumar Jalali.

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Supplementary Fig. 1

Phylogenetic tree of partial 16S rRNA genes showing the cultured bacteria from the guts of different populations of N. lugens constructed by Neighbor-joining method. The numbers at the nodes represent bootstrap values out of 1000 replications. (GIF 56 kb)

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Malathi, V.M., Jalali, S.K., Lyju, V.J. et al. Associated bacterial diversity of insecticide-susceptible and -resistant brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae) analyzed by culture-dependent and -independent methods. Phytoparasitica 45, 683–693 (2017). https://doi.org/10.1007/s12600-017-0629-3

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