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Expression of Brassica oleracea FtsZ1-1 and MinD alters chloroplast division in Nicotiana tabacum generating macro- and mini-chloroplasts

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Abstract

FtsZ1-1 and MinD plastid division-related genes were identified and cloned from Brassica oleracea var. botrytis. Transgenic tobacco plants expressing BoFtsZ1-1 or BoMinD exhibited cells with either fewer but abnormally large chloroplasts or more but smaller chloroplasts relative to wild-type tobacco plants. An abnormal chloroplast phenotype in guard cells was found in BoMinD transgenic tobacco plants but not in BoFtsZ1-1 transgenic tobacco plants. Transgenic tobacco plants bearing the macro-chloroplast phenotype had 10 to 20-fold increased levels of total FtsZ1-1 or MinD, whilst the transgenic tobacco plants bearing the mini-chloroplast phenotype had lower increased FtsZ1-1 or absence of detectable MinD. We also described for the first time, plastid transformation of macro-chloroplast bearing tobacco shoots with a gene cassette allowing for expression of green fluorescent protein (GFP). Homoplasmic plastid transformants from normal chloroplast and macro-chloroplast tobacco plants expressing GFP were obtained. Both types of transformants accumulated GFP at ~6% of total soluble protein, thus indicating that cells containing macro-chloroplasts can regenerate shoots in tissue culture and can stably integrate and express a foreign gene to similar levels as plant cells containing a normal chloroplast size and number.

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Abbreviations

Arc :

Accumulation and replication of chloroplasts

CTAB:

Cetyl trimethylammonium bromide

EST:

Expressed sequence tag

MEGA:

Molecular evolutionary genetics analysis

MS:

Murashige and Skoog

rbcS:

Rubisco small sub-unit

TIGR:

The Institute for Genomic Research

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Acknowledgments

Antibodies for AtFtsZ1-1 and AtMinD were generously provided by Prof. Katherine Osteryoung (Michigan State University, USA) and by Dr. Shin-ya Miyagishima (RIKEN, Japan), respectively.

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  1. School of Applied Sciences, RMIT University, Plenty Road, Bundoora, VIC, 3083, Australia

    Veera R. N. Chikkala, Gregory D. Nugent, David M. Stalker, Aidyn Mouradov & Trevor W. Stevenson

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  1. Veera R. N. Chikkala
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  2. Gregory D. Nugent
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Correspondence to Gregory D. Nugent.

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Communicated by R. Rose.

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Chikkala, V.R.N., Nugent, G.D., Stalker, D.M. et al. Expression of Brassica oleracea FtsZ1-1 and MinD alters chloroplast division in Nicotiana tabacum generating macro- and mini-chloroplasts. Plant Cell Rep 31, 917–928 (2012). https://doi.org/10.1007/s00299-011-1212-x

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