Abstract
Phytocystatins are inhibitors of cysteine-proteases from plants putatively involved in plant defence based on their capability of inhibit heterologous enzymes. We have previously characterised the whole cystatin gene family members from barley (HvCPI-1 to HvCPI-13). The aim of this study was to assess the effects of barley cystatins on two phytophagous spider mites, Tetranychus urticae and Brevipalpus chilensis. The determination of proteolytic activity profile in both mite species showed the presence of the cysteine-proteases, putative targets of cystatins, among other enzymatic activities. All barley cystatins, except HvCPI-1 and HvCPI-7, inhibited in vitro mite cathepsin L- and/or cathepsin B-like activities, HvCPI-6 being the strongest inhibitor for both mite species. Transgenic maize plants expressing HvCPI-6 protein were generated and the functional integrity of the cystatin transgene was confirmed by in vitro inhibitory effect observed against T. urticae and B. chilensis protein extracts. Feeding experiments impaired on transgenic lines performed with T. urticae impaired mite development and reproductive performance. Besides, a significant reduction of cathepsin L-like and/or cathepsin B-like activities was observed when the spider mite fed on maize plants expressing HvCPI-6 cystatin. These findings reveal the potential of barley cystatins as acaricide proteins to protect plants against two important mite pests.
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Acknowledgments
We are grateful to Dr. Vicente Marco (Universidad de la Rioja, España) and Dr. Marina Gambardella (Pontificia Universidad Católica de Chile, Chile) for providing the colonies of T. urticae and the B. chilensis lyophilized samples, respectively. We thank Dr. Christou and Dr. Capell from the University of Lleida (Spain) for their support on the maize stable transformation. The financial support from the Spanish Ministerio de Ciencia e Innovación (project BFU2008-01166), the Agencia Española de Cooperación Internacional para el Desarrollo (Project A/023680/09) and the Spanish Ministerio de Medioambiente y Medio Rural y Marino is gratefully acknowledged.
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Communicated by Q. Zhao.
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Carrillo, L., Martinez, M., Ramessar, K. et al. Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases. Plant Cell Rep 30, 101–112 (2011). https://doi.org/10.1007/s00299-010-0948-z
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DOI: https://doi.org/10.1007/s00299-010-0948-z