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Molecular identification and characterization of the Asian citrus psyllid vector, Diaphorina citri (Hemiptera: Psyllidae) and the transmitted Huanglongbing-associated bacterium, Candidatus Liberibacter asiaticus in India

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Abstract

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the vector of Candidatus Liberibacter asiaticus (CLas), a proteobacterium associated with the most lethal Huanglongbing (HLB) or citrus greening disease. This study was aimed to detect CLas in naturally occurring D. citri vectors and to find out the genetic interconnection and diversity among the populations of D. citri and CLas in the citrus belts of India. Morphological and molecular identification confirmed the 561 adult psyllid samples, collected from 32 citrus orchards located in 8 states of India, as D. citri. Sequence data of two different genes, cytochrome oxidase subunit I (mtCOI), and 16S rRNA, were used to characterize the populations of D. citri and CLas, respectively. In the mtCOI-based phylogenetic tree, two major genetic clusters were observed (A and B) and further in the cluster B, four genetic subgroups (B1, B2, B3 and B4) were noticed. The tree topology clearly indicated that D. citri mtCOI sequences obtained from eight different Indian states separated into four different clades: A, B1, B2 and B4, composed of mtCOI haplotypes Dcit-6, Dcit-2, Dcit-23 and Dcit-1, respectively. Presence of CLas was also validated in the psyllid vector through quantitative real time PCR, conventional PCR and phylogenetic analysis of 16S rRNA gene sequences. These findings may be helpful in the development of region-wide management strategy to be adopted for controlling the D. citri and/or the HLB disease it vectors in different citrus production zones of India.

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Acknowledgements

We thank all the state government staff for their generous help in conducting survey and sample collection work and Dr. M. S. Ladaniya, Director, Central Citrus Research Institute, Nagpur for his encouragement. Financial assistance from ICAR, New Delhi is duly acknowledged.

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  1. ICAR-Central Citrus Research Institute, Amravati Road, Nagpur, 440033, India

    Ashis K. Das, Chandakanna Nandikeshwar Rao, Anjitha George & Shubham A. Chichghare

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  1. Ashis K. Das
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Das, A.K., Rao, C.N., George, A. et al. Molecular identification and characterization of the Asian citrus psyllid vector, Diaphorina citri (Hemiptera: Psyllidae) and the transmitted Huanglongbing-associated bacterium, Candidatus Liberibacter asiaticus in India. J Plant Pathol 104, 1097–1110 (2022). https://doi.org/10.1007/s42161-022-01155-6

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