Abstract
In Arabidopsis leaf coloration mutants, the delayed greening phenomenon is common. Nonetheless, the mechanism remains largely elusive. Here, a delayed greening mutant fln2–4 of FLN2 (Fructokinase-Like Protein2) was studied. FLN2 is one component of Transcriptionally Active Chromosome (TAC) complex which is thought to contain the complete plastid-encoded polymerase (PEP). fln2–4 displayed albino phenotype on medium without sucrose. The PEP-dependent plastid gene expression and chloroplast development were inhibited in fln2–4. Besides interacting with thioredoxin z (TRX z), we identified that FLN2 interacted with another two members of TAC complex in yeast including its homologous protein FLN1 (Fructokinase-Like Protein1) and pTAC5. This indicates that FLN2 functions in regulation of PEP activity associated with these TAC components. fln2–4 exhibited delayed greening on sucrose-containing medium. Comparison of the PEP-dependent gene expression among two complete albino mutants (trx z and ptac14), two yellow mutants (ecb2–2 and ys1) and the fln2–4 showed that fln2–4 remains partial PEP activity. FLN2 and FLN1 are the target proteins of TRX z involved in affecting the PEP activity. Together with the data that FLN1 could interact with itself in yeast, FLN1 may form a homodimer to replace FLN1–FLN2 as the TRX z target in redox pathway for maintaining partial PEP activity in fln2–4. We proposed the partial PEP activity in the fln2 mutant allowed plastids to develop into fully functional chloroplasts when exogenous sucrose was supplied, and finally the mutants exhibited green phenotype.
Highlights
The first step in the life cycle of a plant is seedling establishment after seed germination
Transmission electron microscopy (TEM) observations revealed that the chloroplasts in the 7-day-old fln2–4 mutants had a visible change in ultrastructural organization with irregular morphology and lacked internal membrane structures (Figure 1D)
To confirm that the knockout of FLN2 was responsible for the defects in the fln2–4 phenotype, a construct containing the genomic sequence of the FLN2 gene, as well as 1517-base pair upstream and the FLAG sequence was introduced into the heterozygous plant (FLN2/fln2–4)
Summary
The first step in the life cycle of a plant is seedling establishment after seed germination. When seedlings are exposed to light, they synthesize chlorophyll; the cotyledons expand and turn green [1]. During this process, the etioplasts of cotyledons develop into photosynthetically functional chloroplasts, which allow seedlings to become photoautotrophy and no longer depend on energy stored in the seed [2,3]. PEP and its associated proteins/DNA can be purified by different biochemical purification procedures [10,11] This protein/DNA-complex, termed ‘‘transcriptionally active chromosomes’’ (TAC), can be separated from the plastid components by gel filtration [12,13,14]. In Arabidopsis and mustard (Sinapis alba), a total of 35 components have been identified in the TAC complex including the core subunits of PEP encoded by plastome-located rpo genes and nuclear encoded subunits [15]
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