Purification of a ribosome-inactivating protein with antioxidation and root developer potencies from Celosia plumosa

Abstract

Considering Celosia plumosa as a potent antiviral plant, the attempt was made to determine, purify and characterize its proteinaceous antiviral elements against tobacco mosaic virus hypersensitive response on Nicotiana glutinosa. By using 60% ammonium sulphateprecipitation, FPLC-based anion and cation-exchange chromatography in 10 and 50 mM NaCl, size-exclusion chromatography in 50 mM NaCl and SDS-PAGE 10%, a 25 kD antiviral protein with ribosome-inactivating/28S rRNase ability was purified from the leaves of C. plumosa at vegetative growth stage. The purified protein showed FRAP-based antioxidant activity in vitro and caused 1.7fold and 1.4-fold increases in the growth rate of root system upon carborundum-based application on the root growth medium of N. glutinosa. The present work reports an antiviral protein with ribosome-inactivating, antioxidation and root developer potencies in C. plumosa as an edible or ornamental plant that may be useful in health and agricultural biotechnology in the future. Keywords Antioxidant Á Antiviral Á Celosia plumosa Á Ribosome inactivating Á Root growth Abbreviations FRAP Ferric reducing antioxidant power AVP Antiviral proteins RIP Ribosome-inactivating proteins TMV Tobacco mosaic virus BME Beta mercaptoethanol PVP Polyvinylpyrrolidone FPLC Fast performance liquid chromatography SDS Sodium dodecyl sulphate EGTA Ethylene glycol tetraacetic acid DTT Dithiothreitol BSA Bovine serum albumin

Antiviral Activity of Ribosome-Inactivating Proteins

“…In many cases, the test has involved applying a RIP solution on the leaf surface of the plant together with the virus and comparing the result with the control that does not contain RIP. In some cases, the virus is applied simultaneously [ 86 , 92 , 113 ] and in others, sometime after the application of the RIP [ 90 , 115 ]. The evaluation of antiviral activity has been done by counting the number of lesions [ 88 , 93 ], the time of onset of symptoms [ 77 , 79 ], the number of infected plants [ 105 ], or the severity of the infection symptoms [ 78 , 115 ].…”

Plant-derived ribosome-inactivating proteins involved in defense against plant viruses

“…The CCP-25 was also reported to inhibit translation of BMV and pokeweed mosaic virus (PkMV) RNA genome in vitro (Baranwal et al, 2002). Another antiviral RIP from C. plumosa at the vegetative stage was reported to exhibit antioxidant activity and inhibited TMV infection (Gholizadeh, 2019).…”

“…RIPs (e.g., PAP, αMMC, BAP-24, BAP-28, CCP-25) were found to be involved in restricting the virus infections through modulation of reactive oxygen species (ROS) as well as altering host cell metabolism to maintain its antioxidant status in plant cells (Bhatia et al, 2004;Yang et al, 2018;Zhu et al, 2016). RIPs exhibit the antioxidant potential that may be involved in the antiviral activity (Gholizadeh, 2019;Gholizadeh et al, 2004). But it has been observed that RIP may reduce the oxidative damage in some way that is independent of antioxidant capacity (Yang et al, 2016).…”