Phenylalanine ammonia-lyase inhibitors do not prevent russet spotting lesion development in lettuce midribs

Abstract

Russet spotting is a physiological disorder of lettuce (Lactuca sativa L.) caused by exposure to hormonal levels (<1 μL · L-1) of ethylene in air at ≃5 °C. Enhanced phenolic metabolism and the accumulation of phenolic compounds accompany the appearance of brown, oval lesions on the leaf midrib. Phenylalanine ammonia-lyase (PAL) is the first committed enzyme in the phenylpropanoid pathway. Three inhibitors of PAL activity [2-aminoindan-2-phosphonic acid (AIP), (α-aminooxyacetic acid (AOA), and (α-aminooxi-β-phenylpropionic acid (AOPP)] greatly reduced the accumulation of phenolic compounds and browning of lesions. At a concentration of 50 μM, AIP inhibited the formation of chlorogenic and dicaffeoyl tartaric acids in cut midribs of iceberg lettuce by 92% and 98%, respectively. AIP competitively inhibited PAL activity from a lettuce mid rib homogenate with an apparent K1 of 22 nM. While the formation of phenolic compounds was strongly inhibited by AIP, the number of lesions associated with russet spotting was not affected. Only the color of the lesions was affected by AIP. In control midribs the russet spotting lesions were brown while those in the AIP-treated midribs were initially olive green and after 3 to 7 days these lesions turned the characteristic brown color. No tyrosine ammonia-lyase activity was detected in a homogenate of lettuce midrib tissue. These results indicate that the early development of russet spotting lesions is independent of the increase in PAL activity and phenolic compounds rather than an effect of these increases as previously suggested. However, accumulation of phenolic compounds does contribute to the subsequent browning symptoms indicative of russet spotting.