Planta

, Volume 230, Issue 1, pp 73–84 | Cite as

Developmentally linked changes in proteases and protease inhibitors suggest a role for potato multicystatin in regulating protein content of potato tubers

  • Sarah M. Weeda
  • G. N. Mohan Kumar
  • N. Richard Knowles
Original Article

Abstract

The soluble protein fraction of fully developed potato (Solanumtuberosum L.) tubers is dominated by patatin, a 40 kD storage glycoprotein, and protease inhibitors. Potato multicystatin (PMC) is a multidomain Cys-type protease inhibitor. PMC effectively inhibits degradation of patatin by tuber proteases in vitro. Herein we show that changes in PMC, patatin concentration, activities of various proteases, and their gene expression are temporally linked during tuber development, providing evidence that PMC has a role in regulating tuber protein content in vivo. PMC was barely detectable in non-tuberized stolons. PMC transcript levels increased progressively during tuberization, concomitant with a 40-fold increase in PMC concentration (protein basis) as tubers developed to 10 g fresh wt. Further increases in PMC were comparatively modest (3.7-fold) as tubers developed to full maturity (250 g). Protease activity declined precipitously as PMC levels increased during tuberization. Proteolytic activity was highest in non-tuberized stolons and fell substantially through the 10-g fresh wt stage. Cys-type proteases dominated the pre-tuberization and earliest stages of tuber development. Increases in patatin transcript levels during tuberization were accompanied by a notable lag in patatin accumulation. Patatin did not begin to accumulate substantially on a protein basis until tubers had reached the 10-g stage, wherein protease activity had been inhibited by approximately 60%. These results indicate that a threshold level of PMC (ca. 3 µg tuber−1, 144 ng mg−1 protein) is needed to favor patatin accumulation. Collectively, these results are consistent with a role for PMC in facilitating the accumulation of proteins in developing tubers by inhibiting Cys-type proteases.

Keywords

Potato multicystatin Protein Protease Tuber development Solanumtuberosum Patatin Tuberization 

Abbreviations

Cys

Cysteine

DAP

Days after planting

EIA

Enzyme ImmunoAssay

FITC

Fluorescein isothiocyanate

LAH

Lipolytic acyl hydrolase

NPM

p-Nitrophenyl myristate

PCPI

Potato cysteine protease inhibitor

PI-1

Potato protease inhibitor I

PI-2

Potato protease inhibitor II

PKPI

Potato Kunitz-type protease inhibitor

PMC

Potato multicystatin

pNPP

p-Nitrophenyl phosphate

Ser

Serine

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Sarah M. Weeda
    • 1
  • G. N. Mohan Kumar
    • 1
  • N. Richard Knowles
    • 1
  1. 1.Postharvest Physiology and Biochemistry Laboratory, Department of Horticulture and Landscape ArchitectureWashington State UniversityPullmanUSA

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