Expression of Chitinase and shRNA Gene Exhibits Resistance to Fungi and Virus

Samia Parveen; Anwar Khan; Nusrat Jahan; Khadija Aaliya; Adnan Muzaffar; Bushra Tabassum; Syed Inayatullah; Syed Moeezullah; Muhammad Tariq; Zainia Rehmat; Niaz Ali; Abrar Hussain

doi:10.3390/genes14051090

Expression of Chitinase and shRNA Gene Exhibits Resistance to Fungi and Virus

Genes (Basel). 2023 May 15;14(5):1090. doi: 10.3390/genes14051090.

Authors

Affiliations

¹ Department of Biotechnology, Balochistan University of Information Technology, Engineering and Management Sciences, Quetta 87300, Pakistan.
² Department of Microbiology, Balochistan University of Information Technology, Engineering and Management Sciences, Quetta 87300, Pakistan.
³ Center of Excellence in Molecular Biology, University of the Punjab, Lahore 54000, Pakistan.
⁴ School of Biological Sciences, University of the Punjab, Lahore 54000, Pakistan.
⁵ Department of Biotechnology, Sardar Bahadur Khan Women's University Balochistan, Quetta 87300, Pakistan.
⁶ Department of Botany, Hazara University, Mansehra 21300, Pakistan.

Abstract

With the increasing global population, saving crops from diseases caused by different kinds of bacteria, fungi, viruses, and nematodes is essential. Potato is affected by various diseases, destroying many crops in the field and storage. In this study, we developed potato lines resistant to fungi and viruses, Potato Virus X (PVX) and Potato Virus Y (PVY), by inoculating chitinase for fungi and shRNA designed against the mRNA of the coat protein of PVX and PVY, respectively. The construct was developed using the pCAMBIA2301 vector and transformed into AGB-R (red skin) potato cultivar using Agrobacterium tumefaciens. The crude protein extract of the transgenic potato plant inhibited the growth of Fusarium oxysporum from ~13 to 63%. The detached leaf assay of the transgenic line (SP-21) showed decreased necrotic spots compared to the non-transgenic control when challenged with Fusarium oxysporum. The transgenic line, SP-21, showed maximum knockdown when challenged with PVX and PVY, i.e., 89 and 86%, while transgenic line SP-148 showed 68 and 70% knockdown in the PVX- and PVY-challenged conditions, respectively. It is concluded from this study that the developed transgenic potato cultivar AGB-R showed resistance against fungi and viruses (PVX and PVY).

Keywords: Fusarium oxysporum; shRNA; virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chitinases* / genetics
Fusarium* / genetics
Plants, Genetically Modified / genetics
Potyvirus* / genetics
RNA, Small Interfering / genetics

Substances

RNA, Small Interfering
Chitinases

Supplementary concepts

Fusarium oxysporum

Grants and funding

The authors wish to thank the Higher Education Commission, Pakistan (HEC) for the financial support for the study in form of research grant HEC-TDF02-12.