Prevalence, transmission and molecular characterization of Cotton leaf curl Multan virus infecting hollyhock plants in Pakistan

Mol Biol Rep. 2022 Jun;49(6):5635-5644. doi: 10.1007/s11033-022-07557-0. Epub 2022 May 22.

Abstract

Background: Hollyhock (Alcea rosea) is an ornamental plant belonging to the Malvaceae family and has a remarkable aesthetic and medicinal value. A number of distinct infectious entity including fungi, nematode, bacteria and most importantly both single and double stranded DNA and RNA viruses are reported from infected hollyhock plant. Begomoviruses, the well reputed member of the family Geminiviridae infected the hollyhock recently with a new hollyhock vein yellowing virus and in the present study it infected the hollyhock plant with Cotton leaf curl Multan virus (CLCuMV) which cause the disease of leaf curling.

Methods and results: The symptomatic leaves of the hollyhock plants were collected based on the characteristic symptoms of leaf curling, puckering as well as vein thickening. DNA was extracted by using the recommended 2× CTAB protocol and PCR technique was optimized for the detection of begomovirus followed by sequencing. The data of disease incidence of infection location wise was collected based on the positive results of PCR amplification. Virus free whitefly collected from cotton field and feed on infected hollyhock plant in cage for few days then used for the transmission study of begomovirus on healthy hollyhock plants. Results of PCR amplification indicated that the primers Av/Ac core, Begomo 01/02, and CLCV 01/02 showed the bands of 579 bp, 2.8 kb and 1.1 kb respectively. The betasatellite was amplified by using beta01/02 and CLCuMuBF11/R33, which showed the band of 1400 bp and 481 bp. Disease incidence and Transmission study confirmed the begomovirus in hollyhock plants at molecular level. The sequence obtained with Av/Ac core primers showed the 99% identity with Cotton leaf curl Multan virus-Rajasthan strain and betasatellite primers showed 98% identity with Cotton leaf curl Multan betasatellite.

Conclusion: Hollyhock plants infected by CLCuMV and associated betasatellite has been reported as a possible source of virus inoculum from Pakistan. These findings extend the range of Begomoviruses and betasatellites known to infect A. rosea and highlight this hollyhock species as an important reservoir of agriculturally important Begomoviruses and betasatellites.

Keywords: Alcea rosea; Begomovirus; CLCuMB; CLCuMV; Virus reservoir; Virus transmission.

MeSH terms

  • Begomovirus* / genetics
  • Malvaceae*
  • Pakistan
  • Phylogeny
  • Plant Diseases / genetics
  • Prevalence

Supplementary concepts

  • Cotton leaf curl Multan virus