The Characterization and the Biological Activity of Phytotoxin Produced by Paraphoma radicina

Paraphoma radicina is a new pathogen that causes alfalfa paraphoma root rot (APRR), leading to alfalfa production losses. The resistance levels of 30 alfalfa cultivars to APRR have already been characterized. However, the pathogenic mechanism of P. radicina is still unclear. This study aimed to assess the effects of a crude toxin extracted from P. radicina cell-free culture filtrate (CFCF) on susceptible and resistant cultivars of alfalfa. Meanwhile, the crude toxin components were detected using gas chromatography-mass spectrometry (GC-MS) analysis. CFCF cultured in MEB medium for 14 days and crude toxin extracted by ethyl acetate induced significant phytotoxicity caused the average lesion areas of 5.8 and 3.9 mm(2), respectively, on alfalfa leaves. The crude toxin exhibited resistance to high temperature, as shown by a lesion area of 3.6 mm(2) when treated at 120 °C for 30 min. Different concentrations of the crude toxin in water and MS medium had different effects on susceptible and resistant cultivars. Moreover, the crude toxin affected the plasma membrane, mitochondria, and nuclear membranes of alfalfa root cortical cells. Further, it induced significant phytotoxicity on Sonchus oleraceus L., Capsella bursa-pastoris (Linn.) Medic, and Chenopodium album L. Agropyron cristatum L. (average lesion areas; 11.6, 15.8, 21.4, and 6.2 mm(2), respectively), indicating that the crude toxin of P. radicina is a non-host-selective toxin. GC-MS analysis detected four possible active substances in the toxin (3-hydroxypyridine, 5-methylresorcinol, 3-Hydroxypropionic acid, and 4-Hydroxyphenylethanol). Therefore, this study may provide insight into the pathogenic mechanism of P. radicina to alfalfa.