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Disruption of the psbA gene by the copy correction mechanism reveals that the expression of plastid-encoded genes is regulated by photosynthesis activity

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Abstract

The functional analysis of genes encoded by the chloroplast genome of tobacco by reverse genetics is routine. Nevertheless, for a small number of genes their deletion generates heteroplasmic genotypes, complicating their analysis. There is thus the need for additional strategies to develop deletion mutants for these genes. We have developed a homologous copy correction-based strategy for deleting/mutating genes encoded on the chloroplast genome. This system was used to produce psbA knockouts. The resulting plants are homoplasmic and lack photosystem II (PSII) activity. Further, the deletion mutants exhibit a distinct phenotype; young leaves are green, whereas older leaves are bleached, irrespective of light conditions. This suggests that senescence is promoted by the absence of psbA. Analysis of the transcript levels indicates that NEP (nuclear-encoded plastid RNA polymerase)-dependent plastid genes are up regulated in the psbA deletion mutants, whereas the bleached leaves retain plastid-encoded plastid RNA polymerase activity. Hence, the expression of NEP-dependent plastid genes may be regulated by photosynthesis, either directly or indirectly.

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Abbreviations

PSII:

Photosystem II

NEP:

Nuclear-encoded plastid RNA polymerase

PEP:

plastid-encoded plastid RNA polymerase

IR:

Inverted repeat

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Acknowledgments

This study was supported by research grants from the Ministry of Science and Technology and from the Pakistan Science Foundation (PSF), Islamabad to MSK and from the Grants-in-Aids (17051025) for scientific research to TS.

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Authors and Affiliations

  1. National Institute for Biotechnology and Genetic Engineering (NIBGE), P. O. Box 577, Jhang Road, Faisalabad, Pakistan

    Muhammad Sarwar Khan & Waqar Hameed

  2. Laboratory of Applied Biology, Faculty of Human Environment, Kyoto Prefectural University, Kyoto, 606-8502, Japan

    Mikio Nozoe & Takashi Shiina

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  1. Muhammad Sarwar Khan
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  2. Waqar Hameed
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  3. Mikio Nozoe
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  4. Takashi Shiina
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Correspondence to Muhammad Sarwar Khan.

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Khan, M.S., Hameed, W., Nozoe, M. et al. Disruption of the psbA gene by the copy correction mechanism reveals that the expression of plastid-encoded genes is regulated by photosynthesis activity. J Plant Res 120, 421–430 (2007). https://doi.org/10.1007/s10265-007-0082-3

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