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Zebra chip disease decreases tuber (Solanum tuberosum L.) protein content by attenuating protease inhibitor levels and increasing protease activities

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Abstract

Main conclusion

Zebra chip disease of potato decreases protease inhibitor levels resulting in enhanced serine-type protease activity, decreased protein content and altered protein profiles of fully mature tubers.

Zebra-chip (ZC), caused by Candidatus Liberibacter solanacearum (CLso), is a relatively new disease of potato that negatively affects growth, yield, propagation potential, and fresh and process qualities of tubers. Diseased plants produce tubers with characteristic brown discoloration of vascular tissue accompanied by elevated levels of free amino acids and reducing sugars. Here we demonstrate that ZC disease induces selective protein catabolism in tubers through modulating protease inhibitor levels. Soluble protein content of tubers from CLso-infected plants was 33 % lower than from non-infected plants and electrophoretic analyses revealed substantial reductions in major tuber proteins. Patatin (~40 kDa) and ser-, asp- (22 kDa) and cys-type (85 kDa) protease inhibitors were either absent or greatly reduced in ZC-afflicted tubers. In contrast to healthy (non-infected) tubers, the proteolytic activity in CLso infected tubers was high and the ability of extracts from infected tubers to inhibit trypsin (ser-type) and papain (cys-type) proteases greatly attenuated. Moreover, extracts from CLso-infected tubers rapidly catabolized proteins purified from healthy tubers (40 kDa patatin, 22 kDa protease inhibitors, 85 kDa potato multicystatin) when subjected to proteolysis individually. In contrast, crude extracts from non-infected tubers effectively inhibited the proteolytic activity from ZC-afflicted tubers. These results suggest that the altered protein profile of ZC afflicted tubers is largely due to loss of ser- and cys-type protease inhibitors. Further analysis revealed a novel PMSF-sensitive (ser) protease (ca. 80–120 kDa) in CLso infected tubers. PMSF abolished the proteolytic activities responsible for degrading patatin, the 22 kDa protease inhibitor(s) and potato multicystatin by CLso infected tubers. The disease-induced loss of patatin and protease inhibitors therefore appears to be modulated by ser-type protease(s). The selective catabolism of proteins in ZC-afflicted tubers undoubtedly affects downstream aspects of carbohydrate and amino acid metabolism, which is ultimately reflected by the accumulation of reducing sugars, free amino acids and reduced sprouting capacity.

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Abbreviations

ANOVA:

Analysis of variance

asp:

Aspartic

BAPNA:

N α-benzoyl-l-arginine 4-nitroanilide hydrochloride

cys:

Cysteine

Pep:

Pepstatin

PIN I:

Potato protease inhibitor I

PIN II:

Potato protease inhibitor II

PKPI:

Potato Kunitz-type protease inhibitor

PMC:

Potato multicystatin

ser:

Serine

ZC:

Zebra chip

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Acknowledgments

We thank Dr. Joseph E. Munyaneza (USDA-ARS, Wapato, WA) for providing tuber samples. Financial support was provided by the USDA-ARS, Washington State Potato Commission, and Washington State University Agricultural Research Center to N.R. Knowles.

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  1. Postharvest Physiology and Biochemistry Laboratory, Department of Horticulture, Washington State University, P.O. Box 646414, Pullman, WA, 99164-6414, USA

    G. N. Mohan Kumar, Lisa O. Knowles & N. Richard Knowles

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Kumar, G.N.M., Knowles, L.O. & Knowles, N.R. Zebra chip disease decreases tuber (Solanum tuberosum L.) protein content by attenuating protease inhibitor levels and increasing protease activities. Planta 242, 1153–1166 (2015). https://doi.org/10.1007/s00425-015-2346-9

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