Cytokinins and phytochrome have both been reported to promote chloroplast development, and possible interactions between the two have been suggested We have examined the effects of red light (R) and a cytokinin, benzyladenine (N6-benzylaminopurine; BA), on the levels of four mRNAs coding for chloroplast proteins in Lemna gibba L The amounts of hybridizable RNA coding for both the major chlorophyll a/b-binding protein and for the small subunit of ribulose-1,5-bisphosphate (RuBP) carboxylase decrease to a low level when white-light-grown L gibba plants are placed in the dark We have previously shown that a subsequent R treatment causes a several-fold increase in the levels of these two messages, and this increase is phytochrome-mediated We have now found that addition of submicromolar concentrations of BA to plants kept in total darkness also results in an increase in levels of these two mRNAs Furthermore, BA treatment magnifies the extent of the response to R treatment However, the levels of mRNAs encoding the large subunit of RuBP carboxylase and the 32-kDa herbicide-binding protein, which are both chloroplastsynthesized messages, are not significantly altered by either R or BA treatment during the same time period The relative amount of β-actin mRNA, a nuclear-encoded message for a cytoplasmic protein, is also not altered either by R or BA treatment Thus, BA treatment does not simply alter the proportion of mRNA to total RNA This conclusion is also supported by the observation that levels of mRNA hybridizing to a sequence abundant in dark-treated plants are not altered by BA treatment The amplification by BA of the R-induced increase in the level of chlorophyll a/b-binding protein mRNA, consistently seen in total RNA, is not observed in RNA isolated from nuclei from plants receiving the same treatments We therefore suggest that cytokinin is regulating expression of this message at a post-transcriptional level, possibly by affecting the stability of the RNA

Benzyladenine modulation of the expression of two genes for nuclear-encoded chloroplast proteins in Lemna gibba: Apparent post-transcriptional regulation.

The effects of a cytokinin (N6-benzyladenine, BA) and light on plastogenesis have been studied in detached Cucurbita cotyledons using the key enzyme of photosynthetic CO2 fixation, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase), as an example of a coordinated program of plastid and nucleo-cytoplasmic gene expression. Treatment of etiolated cotyledons with either BA in darkness or in light or light alone results in a marked and correlated stimulation of enzyme activity, quantity and biosynthesis (in-vivo [14C]leucine incorporation into immunoprecipitated enzyme protein), indicating an increase of de-novo synthesis under the influence of the two effectors. Cell-free translation of non-polyadenylated (poly(A)-)RNA in an Escherichia coli system and total RNA in a wheat-germ system likewise demonstrate a light and hormone-dependent increase in the amounts of translatable mRNAs for the large (LS) and small subunits (SS) of RuBPCase (among other polypeptides). Hybridisation of poly(A)-RNA with a nick-translated LS gene of spinach RuBPCase reveals also two-to three-fold BA-or light-induced enhancement of LS mRNA content. Indications for stimulation of SS mRNA transcription are derived from inhibitor experiments with cordycepin. We conclude that the observed stimulation of de-novo RuBPCase synthesis by cytokinin and by light can be referred to the level of mRNA transcription, but it remains open whether this is a primary action. Furthermore, our results indicate that the coaction of the two exogenous factors is additive at different steps of RuBPCase formation, indicating independent actions in the causal chain between effector-signal transduction and RuBPCase gene expression.

Gene expression in cytokinin-and light-mediated plastogenesis of Cucurbita cotyledons: ribulose-1,5-bisphosphate carboxylase/oxygenase.

Cytokinin has been reported to mimic some of the effects of light on de-etiolation responses in dark-grown Arabidopsis seedlings. The interaction between cytokinin and light was examined by analyzing cytokinin dose and light fluence effects on hypocotyl elongation in wild-type and mutant Arabidopsis seedlings with defects in light or hormone responses. It was found that (a) cytokinin and light-response systems have independent and additive effects on the inhibition of hypocotyl elongation and (b) either cytokinin or light can saturate the morphogenic responses. As a consequence, cytokinin has no effect on hypocotyl elongation under normal growth conditions because light levels saturate the hypocotyl inhibition response. To determine whether a functional light-response pathway is required for cytokinin responses, light-insensitive long hypocotyl (hy) mutants were tested for cytokinin responses. The hy mutants (hy1 to hy6) had normal cytokinin responses, except phyB-1 (hy3-1), in which hypocotyl elongation was insensitive to cytokinin. Cytokinin insensitivity in phyB-1 was attributed to an indirect effect of the mutation on cytokinin responses. The effects of cytokinin on the inhibition of hypocotyl elongation are largely mediated by ethylene, and blocking the ethylene-response pathway through the action of a cytokinin-resistant, ethylene-insensitive mutant (ckr1/ein2) had no effect on the light inhibition of hypocotyl elongation. These results do not support the idea that cytokinin mediates the action of light on hypocotyl elongation.

The Effects of Cytokinin and Light on Hypocotyl Elongation in Arabidopsis Seedlings Are Independent and Additive.

Northern blot analysis revealed that a single 4.2 kb phytochrome mRNA species was detectable in cotyledons excised from five-day-old etiolated cucumber seedlings. Intact etiolated five-day-old cucumber seedlings were given a red light or benzyladenine treatment, and cotyledons were harvested at various times following treatment. The abundance of phytochrome mRNA in the cotyledons was quantitated using 32P-labeled RNA probes and slot blot analysis. By 2 h after irradiation the phytochrome mRNA level was reduced to 40% of the initial abundance and reaccumulation began by 3 h after irradiation. Reaccumulation of phytochrome mRNA to the time-zero dark control level was achieved by 10 h after treatment. A decrease in phytochrome mRNA abundance was evident by 2 h after benzyladenine treatment, and a maximal reduction to 45% of the time-zero dark control was attained by 4 h after treatment. No recovery of the phytochrome mRNA level was evident by 8 h after benzyladenine treatment. The abundance of actin mRNA was unaffected by benzyladenine treatment.

Down-regulation of phytochrome mRNA abundance by red light and benzyladenine in etiolated cucumber cotyledons

A reduced red to far-red (R/FR) light ratio and low photosynthetically active radiation (PAR) irradiance are both strong signals for inducing etiolation growth of plant stems. Under natural field conditions, plants can be exposed to either a reduced R/FR ratio or lower PAR, or to a combination of both. We used Helianthus annuus L., the sunflower, to study the effect of reduced R/FR ratio, low PAR or their combination on hypocotyl elongation. To accomplish this, we attempted to uncouple light quality from light irradiance as factors controlling hypocotyl elongation. We measured alterations in the levels of endogenous gibberellins (GAs), cytokinins (CKs) and the auxin indole-3-acetic acid (IAA), and the effect of exogenous hormones on hypocotyl growth. As expected, both reduced R/FR ratio and lower PAR can significantly promote sunflower hypocotyl elongation when given separately. However, providing the reduced R/FR ratio at a low PAR resulted in the greatest hypocotyl growth, and this was accompanied by significantly higher levels of endogenous IAA, GA1, GA8, GA20 and of a wide range of CKs. Providing a reduced R/FR ratio under normal PAR also significantly increased growth and again gave significantly higher levels of endogenous IAA, GAs and CKs. However, only under the de-etiolating influence of a normal R/FR ratio did lowering PAR significantly increase levels of GA1, GA8 and GA20. We thus conclude that light quality (e.g. the R/FR ratio) is the most important component of shade for controlling hypocotyl growth and elevated growth hormone content.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-3040.2006.01612.x

The interaction of light quality and irradiance with gibberellins, cytokinins and auxin in regulating growth of Helianthus annuus hypocotyls

Intact mustard seedlings were treated with zeatin and photomorphogenetically active light in different ways: (1) hormone treatment preceding light treatment, (2) light treatment preceding hormone treatment, (3) hormone and light applied simultaneously. Under all experimental conditions the effect of the hormone treatment is multiplicative to the light effect with regard to the increase of cotyledon area. However, the hormone effect is additive to the light effect with regard to increases of the level of NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) and carotenoid contents. Anthocyanin synthesis is inhibited by exogenous zeatin whereby the concentration response curves are similar, irrespective of the extent of anthocyanin formation mediated by light. However, an interaction was found in the sense that the responsiveness toward zeatin is decreased somewhat by the action of phytochrome. Our results show that the responsiveness to light (via the far-red-absorbing form of phytochrome; P
fr) is not changed by a preceding or simultaneous hormone treatment. Moreover, the responsiveness of the plant to exogenously applied zeatin is not affected — except in anthocyanin synthesis — by a preceding or simultaneous light treatment. We conclude from our results that the action of phytochrome on the developmental processes is not related to cytokinin levels.

Zhe Tong

Papers

Coaction of light and cytokinin in photomorphogenesis.