Lignin is involved in the reduction of Verticillium dahliae var. longisporum inoculum in soil by crop residue incorporation

Verticillium wilt is an increasing problem in European cauliflower production. In this study, several crop residues were screened for their ability to reduce the viability of microsclerotia when incorporated into soil. In addition, the role of fungitoxic volatiles and lignin in the crop residue-mediated reduction in microsclerotia viability was studied. Broccoli (Brassica oleracea var. italica), cauliflower (B. oleracea var. botrytis), Indian mustard (Brassica juncea), ryegrass (Lolium perenne) and corn (Zea mays) were incorporated in naturally infested soil samples collected from two cauliflower fields in Belgium, labelled Is1 and S3. The effectiveness in reducing the viability of microsclerotia depended on the soil sample and on the type of residue. In the Is1 soil, broccoli, cauliflower and ryegrass incorporation significantly reduced the inoculum level by more than 90%, while Indian mustard significantly reduced numbers of viable microsclerotia by 50%. In the S3 soil, broccoli, cauliflower and Indian mustard were not effective, whereas ryegrass and corn incorporation reduced the microsclerotia level by 50% or more. In conclusion, incorporation of ryegrass and corn was more effective than incorporation of crucifer residues. In the conditions tested, fungicidal volatile compounds did not play an important role in Verticillium microsclerotia reduction in soil. Volatiles from broccoli and cauliflower did not affect microsclerotia viability in an in vitro bioassay, whereas the volatiles from Indian mustard killed the microsclerotia. Indian mustard incorporation in soil, however, only had a minor effect on microsclerotia viability. In the S3 soil, 1% (w/w) Kraft pine lignin, a waste product of the paper industry, had to be added to observe a significant reduction on the viability of microsclerotia, whereas in the Is1 soil, a significant effect was observed when 0.1% (w/w) Kraft pine lignin was added. Acid-insoluble lignin was extracted from all crop residues previously tested. Crop residues with high lignin content seemed to be more effective than crop residues with low lignin content. The reduction of Verticillium microsclerotia viability depended on lignin type and on crop structure, since lignin extracted from cauliflower leaves was more effective than lignin extracted from cauliflower stems and corn leaves were more effective than corn roots. Microsclerotia reduction was higher after fresh residue incorporation than after incorporation of their extracted acid-insoluble lignin, indicating that the effect of crop residue incorporation on microsclerotia viability cannot be explained solely by the effects of lignin. Incorporation of lignin-rich substrates in soil may open up new perspectives for integrated control of Verticillium.