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Published on June 30, 2006; 10.1104/pp.106.082404

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Received April 20, 2006
Accepted June 21, 2006

Expression and functional analyses of the plastid lipid-associated protein CHRC suggest its role in chromoplastogenesis and stress

Yael Leitner-Dagan , Marianna Ovadis , Elena Shklarman , Yigal Elad , Dalia Rav David , and Alexander Vainstein *

The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel
Department of Plant Pathology and Weed Research, ARO, The Volcani Center, Bet Dagan, 50250 Israel

* Corresponding author; email: vain{at}agri.huji.ac.il.

Chromoplastogenesis during flower development and fruit ripening involves the dramatic overaccumulation of carotenoids sequestered into structures containing lipids and proteins termed PAPs (plastid lipid-associated proteins). CHRC, a cucumber (Cucumis sativus) PAP, has been suggested to be transcriptionally activated in carotenoid-accumulating flowers by gibberellic acid (GA). Mybys, a MYB-like trans-activator identified here, may represent a chromoplastogenesis-related factor: its expression is flower-specific and parallels that of ChrC during flower development; moreover, as revealed by stable ectopic and transient-expression assays, it specifically trans-activates ChrC promoter in flowers accumulating carotenoids and flavonoids. A detailed dissection of ChrC promoter revealed a GA-responsive element, gacCTCcaa, the mutation of which abolished ChrC activation by GA. This cis-element is different from the GARE motif and is involved in ChrC activation probably via negative regulation, similar to other GA-responsive systems. The GA responsiveness and MYBYS floral activation of the ChrC promoter do not overlap with respect to cis-elements. To study the functionality of CHRC, which is activated in vegetative tissues similar to other PAPs by various biotic and abiotic stresses, we employed a tomato plant system and generated RNAi-transgenic lines with suppressed LeCHRC. Transgenic flowers accumulated ca. 30% less carotenoids per unit protein than controls, indicating an interrelationship between PAPs and flower-specific carotenoid accumulation in chromoplasts. Moreover, the transgenic LeCHRC-suppressed plants were significantly more susceptible to Botrytis cinerea infection, suggesting CHRC's involvement in plant protection under stress conditions and supporting the general, evolutionarily preserved role of PAPs.






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