Abstract
Phototropins are plant photoreceptors which regulate numerous responses to blue light, including chloroplast relocation. Weak blue light induces chloroplast accumulation, whereas strong light leads to an avoidance response. Two Arabidopsis phototropins are characterized by different light sensitivities. Under continuous light, both can elicit chloroplast accumulation, but the avoidance response is controlled solely by phot2. As well as continuous light, brief light pulses also induce chloroplast displacements. Pulses of 0.1s and 0.2s of fluence rate saturating the avoidance response lead to transient chloroplast accumulation. Longer pulses (up to 20s) trigger a biphasic response, namely transient avoidance followed by transient accumulation. This work presents a detailed study of transient chloroplast responses in Arabidopsis. Phototropin mutants display altered chloroplast movements as compared with the wild type: phot1 is characterized by weaker responses, while phot2 exhibits enhanced chloroplast accumulation, especially after 0.1s and 0.2s pulses. To determine the cause of these differences, the abundance and phosphorylation levels of both phototropins, as well as the interactions between phototropin molecules are examined. The formation of phototropin homo- and heterocomplexes is the most plausible explanation of the observed phenomena. The physiological consequences of this interplay are discussed, suggesting the universal character of this mechanism that fine-tunes plant reactions to blue light. Additionally, responses in mutants of different protein phosphatase 2A subunits are examined to assess the role of protein phosphorylation in signaling of chloroplast movements.
Highlights
Gateway entry vector with 35S promoter used for multisite gateway reaction
Gateway entry vector with nGFP used for multisite gateway reaction to obtain nGFP fusion at the N-terminus of the protein
Gateway entry vector with cGFP used for multisite gateway reaction to obtain cGFP fusion at the N-terminus of the protein
Summary
Nucleotide sequence (5' - 3') AAACATAGCCACACGCATTTC GGCCAGCCAGTTAGGTATAGG TCCAAGAATTCACCATTTTGG GAGGTCCCGAGTTCAATTCTC TGGTTCACGTAGTGGGCCATCG Primer name attb1_phot1_for attb2_phot1_rev attb2_phot1_rev_nostop attb1_phot2_f attb2_phot2_r attb2_phot2_rev_nostop attB1_phot1_Cterm_for attB2_phot1_Nterm_rev attB1_phot2_Cterm_for attB2_phot2_Nterm_rev Nucleotide sequence (5' - 3') GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGGAACCAACAGAAAAACC GGGGACCACTTTGTACAAGAAAGCTGGGTCAAAAAAACATTTGTTTGCAG GGGGACCACTTTGTACAAGAAAGCTGGGTCAAAAACATTTGTTTGCAGATC GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGGAGAGAGGCCAAGAGCC GGGGACCACTTTGTACAAGAAAGCTGGGTCTTAGAAGAGGTCAATGTCCAAG GGGGACCACTTTGTACAAGAAAGCTGGGTCGAAGAGGTCAATGTCCAAGTC GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGACACCAGAGGATTTATGGGC GGGGACCACTTTGTACAAGAAAGCTGGGTCGTTGGCATCAGGAAGTTCTCGAAC GGGGACAAGTTTGTACAAAAAAGCAGGCTCGCATCATTTCAAACCAATAAAACC GGGGACCACTTTGTACAAGAAAGCTGGGTCTAGTCCCACTGTTTCTCCACTC
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