Generation and characterization of a collection of knock-down lines for the chloroplast Clp protease complex in tobacco
| Autor: | Michael Tillich, Mercedes Diez, Mark Aurel Schöttler, Ralph Bock, Nadine Tiller, Juan Moreno, Daniel Karcher |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Nicotiana Proteases Physiology medicine.medical_treatment Protein subunit Plant Science Biology Protein degradation 01 natural sciences Chloroplast 03 medical and health sciences RNA interference Tobacco Clp protease medicine Plastid tabacum plastid Metalloproteinase Protease photosynthesis fungi protein stability food and beverages Metalloendopeptidases protease Endopeptidase Clp Molecular biology Cell biology 030104 developmental biology Gene Knockdown Techniques Mutagenesis Site-Directed protein degradation RNA Interference leaf development 010606 plant biology & botany Research Paper |
| Zdroj: | Journal of Experimental Botany |
| ISSN: | 1460-2431 |
| Popis: | Highlight By generating a large set of transgenic and transplastomic knock-down lines in tobacco, the phenotypic effects of impaired chloroplast protease function on plant growth and development, leaf morphology, pigmentation, and photosynthesis were characterized. Protein degradation in chloroplasts is carried out by a set of proteases that eliminate misfolded, damaged, or superfluous proteins. The ATP-dependent caseinolytic protease (Clp) is the most complex protease in plastids and has been implicated mainly in stromal protein degradation. In contrast, FtsH, a thylakoid membrane-associated metalloprotease, is believed to participate mainly in the degradation of thylakoidal proteins. To determine the role of specific Clp and FtsH subunits in plant growth and development, RNAi lines targeting at least one subunit of each Clp ring and FtsH were generated in tobacco. In addition, mutation of the translation initiation codon was employed to down-regulate expression of the plastid-encoded ClpP1 subunit. These protease lines cover a broad range of reductions at the transcript and protein levels of the targeted genes. A wide spectrum of phenotypes was obtained, including pigment deficiency, alterations in leaf development, leaf variegations, and impaired photosynthesis. When knock-down lines for the different protease subunits were compared, both common and specific phenotypes were observed, suggesting distinct functions of at least some subunits. Our work provides a well-characterized collection of knock-down lines for plastid proteases in tobacco and reveals the importance of the Clp protease in physiology and plant development. |
| Databáze: | OpenAIRE |
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