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Plant and Cell Physiology Advance Access published online on March 7, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci079
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received September 22, 2004
Accepted February 16, 2005

Regular Paper

The Formation of Zn-Chl a in Chlorella Heterotrophically Grown in the Dark with An Excessive Amount of Zn2+

Isamu Ikegami 1*, Aki Nemoto 1, and Kohki Sakashita 1

1 Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-0195, Japan

* To whom correspondence should be addressed.
Isamu Ikegami, E-mail: i-ike{at}pharm.teikyo-u.ac.jp


  Abstract

Chlorella, when heterotrophically cultivated in the dark, is able to grow with Zn2+ at 10-40 mM, which is ten times the concentration lethal to autotrophically grown cells. However, the lag phase prolongs with increasing concentration of Zn2+; for example, in this study, one day of the control lag phase prolonged to about 16 days with Zn2+ at 16.7 mM (x2,000 of the control). Once the cells started to grow, the log phase was finished within 4-6 days regardless of Zn concentration, which was almost the same as that of the control. The PSI reaction center Chl, P700, and the far-red fluorescence were detected only after the late log phase of the growth curve, suggesting that chlorophyll-protein complexes can be organized after cell division has ceased. Interestingly, at more than 16.7 mM of Zn2+, Zn-Chl a was accumulated and finally accounted for about 25% of the total Chl a in the late stationary phase. We found that the Zn-Chl a was present in the thylakoid membranes and not in the soluble fractions of the cells. The rather low fluorescence yield at around 680 nm in the stationary phase suggests that Zn-Chl a can transfer its excitation energy to other chlorophylls. Before accumulation of Zn-Chl a, a marked amount of pheophytin a was temporally accumulated, suggesting that Zn-Chl a could be chemically synthesized via Phe a.

Keywords: Chlorophyll a; Zn-chlorophyll a; Zinc acclimation; Low temperature fluorescence; Chlorella.
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