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Published on December 12, 2008; 10.1104/pp.108.130013

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Received September 19, 2008
Accepted December 9, 2008

Autophagy plays a role in chloroplast degradation during senescence in individually darkened leaves

Shinya Wada , Hiroyuki Ishida *, Masanori Izumi , Koki Yoshimoto , Yoshinori Ohsumi , Tadahiko Mae , and Amane Makino

Department of Applied Plant Science, Graduate School of Agricultural Sciences, Tohoku University, Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan; RIKEN Plant Science Center, Suehiro-cho 1-7-22, Tsurumi-ku, Yokohama 230-0045, Japan; Department of Cell Biology, National Institute for Basic Biology, Myodaiji-cho, Okazaki 444-8585, Japan

* Corresponding author; email: hiroyuki{at}biochem.tohoku.ac.jp.

Chloroplasts contain approximately 80% of total leaf nitrogen and represent a major source of recycled nitrogen during leaf senescence. While bulk degradation of the cytosol and organelles in plants is mediated by autophagy, its role in chloroplast catabolism is largely unknown. We investigated the effects of autophagy disruption on the number and the size of chloroplasts during senescence. When leaves were individually darkened, senescence was promoted similarly in both wild-type Arabidopsis (Arabidopsis thaliana) and in an autophagy defective mutant, atg4a4b-1. The number and size of chloroplasts decreased in darkened leaves of wild-type, while the number remained constant and the size decrease was suppressed in atg4a4b-1. When leaves of transgenic plants expressing stromal-targeted DsRed were individually darkened, a large accumulation of fluorescence in the vacuolar lumen was observed. Chloroplasts exhibiting chlorophyll fluorescence as well as Rubisco-containing bodies were also observed in the vacuole. No accumulation of stroma-targeted DsRed, chloroplasts, or Rubisco-containing bodies was observed in the vacuoles of the autophagy-deficient mutant. We have succeeded in demonstrating chloroplast autophagy in living cells and provide direct evidence of chloroplast transportation into the vacuole.




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