Transmission, characterization and occurrence of recombination in Indian strain of squash leaf curl China virus associated with yellow mosaic and leaf curl disease of Summer squash

V Venkataravanappa; C N Lakshminarayana Reddy; M Nandan; Shridhar Hiremath; K V Ashwathappa; K S Shankarappa; H D Vinay Kumar; M Krishna Reddy

doi:10.1007/s13205-021-02821-9

Transmission, characterization and occurrence of recombination in Indian strain of squash leaf curl China virus associated with yellow mosaic and leaf curl disease of Summer squash

3 Biotech. 2021 Jun;11(6):265. doi: 10.1007/s13205-021-02821-9. Epub 2021 May 12.

Authors

V Venkataravanappa¹, C N Lakshminarayana Reddy², M Nandan², Shridhar Hiremath², K V Ashwathappa³, K S Shankarappa⁴, H D Vinay Kumar², M Krishna Reddy³

Affiliations

¹ Division of Plant Pathology, CHES, ICAR-Indian Institute of Horticultural Research, Chettalli, Madikeri District, Bangalore, Karnataka, 571248 India.
² Department of Plant Pathology, College of Agriculture, University of Agricultural Sciences, GKVK, Bengaluru, Karnataka 563125 India.
³ Indian Institute of Horticultural Research, Hessaraghatta Lake PO, Bangalore, Karnataka 560089 India.
⁴ Department of Plant Pathology, College of Horticulture, University of Horticultural Sciences, Bagalkot, Bengaluru, Karnataka 560065 India.

Abstract

Summer squash is one of the important vegetable crops and its production is hampered by various abiotic and biotic stresses. Of the different biotic stresses, viral infections are responsible for causing great losses to this crop. Diseases caused by begomoviruses are becoming a major constraint in the cultivation of summer squash. Samples from summer squash plants exhibiting severe yellow mosaic and leaf curl symptoms were collected from the Varanasi district of Uttar Pradesh (India) and begomovirus associated with these plants was transmitted through whiteflies (Bemisia tabaci) to healthy squash plants. The relationship between causal virus and whitefly vector was determined. The minimum acquisition access period (AAP) and inoculation feeding period (IFP) required by B. tabaci to transmit the virus was determined to be 10 min and female insects have greater efficiency in transmitting virus than male insects. The partial genome of the virus was amplified by PCR (1.2 kb), cloned and sequenced from the ten infected plant samples collected from field. Partial genome sequence analysis (1.2 kb) obtained from the ten samples revealed that they are associated with begomovirus species closely related to the Indian strain of Squash leaf curl China virus (SLCCNV). Therefore, one representative sample (Sq-1) was selected and complete genome of the virus was amplified by rolling circle amplification (RCA) method. Sequence analysis by Sequence Demarcation Tool (SDT) showed that the current isolate has maximum nucleotide (nt) identity of 93.7-98.4% and 89-98.1% with respect to DNA A DNA B, respectively with Indian strains of SLCCNV infecting cucurbits in India. Recombination analysis of genomes (DNA A and DNA B components) showed that a major part of genomes likely to be originated from already known begomoviruses (ToLCNDV, SLCCNV-CN and SLCCNV-IN) are infecting cucurbitaceous crops. Serological assays such as triple antibody sandwich-enzyme-linked immune-sorbent (TAS-ELISA) assay, dot blot immunobinding assay (DIBA), immuno-capture polymerase chain reaction (IC-PCR) were developed for the detection of SLCCNV.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-021-02821-9.

Keywords: Begomovirus; Bemisia tabaci; PCR; Phylogenetic analysis; Recombination; Squash; Squash leaf curl China virus.