Dormant, dehulled red rice (Oryza sativa L. ‘Louisiana strawhulled′) germinated 80 to 90% at 30 C upon exposure to 10-3 M kinetin (6-furfurylaminopurine), benzyladenine (6-benzylaminopurine), or isopentenyladenine [6-(3-methylbut-2-enylamino)purine] at pH 3.5. Zeatin [6-(4-hydroxy-3-methylbut-trans-2-enylamino)purine] was also active, eliciting 45% germination at 10-3 M. Adenine was inactive at concentrations as high as 10-2 M. There was little influence of light and no effect of initial pH of the medium upon cytokinin-stimulated germination. At 20 C the response to cytokinins was reduced, but germination of non-dormant seeds was not prevented. At 30 C, the germination of dehulled red rice with 10-3 M kinetin occurred rapidly, with almost 70% germination after 2 days. When dehulled seeds were incubated in water and then transferred to kinetin, percent germination decreased as the duration of water incubation increased. The loss of cytokinin response did not parallel the timecourse of imbibition. Cytokinins did not break dormancy of intact seeds. The hull appeared to limit kinetin uptake and inhibited germination even when kinetin was forced into intact seeds by vacuum infiltration. Dry-after-ripening of intact seeds at 30 C did not increase the kinetin-sensitivity of intact red rice.

Pretreatment of etiolated cucumber cotyledons with potassium (K), benzyladenine (BA), or δ-aminolevulinic acid (ALA) for 20 h in the dark, followed by exposure to light for 24 h, increased chlorophyll and carotenoid content. A similar treatment with fluridone {1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone} decreased the levels of these photosynthetic pigments. However, when the etiolated cotyledons were pretreated with either BA or ALA for 20 h in the dark, followed by fluridone during a 24-h light period, the inhibitory effects of fluridone on chlorophyll content were reversed and the values were higher than the controls. In these treatments, ALA did not counteract the inhibitory effects of fluridone on carotenoids, but BA reduced the effects. In the cucumber cotyledon greening system, K was found to be most effective in stimulating both chlorophyll and carotenoid accumulation. When K was administered to the cotyledons either together with, prior to, or after the fluridone treatment, the inhibitory effect of fluridone on chlorophyll accumulation was totally eliminated and the stimulatory effects due to K were still maintained. Although K increased carotenoid content, it did not reverse the inhibitory effect of fluridone on carotenoids. From these findings it is concluded that the inhibition of chlorophyll and carotenoid accumulation by fluridone may be mediated via unrelated mechanisms at fluridone concentrations that do not totally eliminate or drastically reduce carotenoid content.

Benzyladenine, a synthetic cytokinin, stimulates DNA metabolism during maize germination; apparently such stimulation occurs in two phases, and during the first phase stimulations of DNA synthesis, of protein kinase activity (both α-amanitin repressible), of DNA polymerase and poly ADP-ribosyl polymerase activities are observed. Since the evidence suggested that the effects produced by BA were due, at least partially, to transcriptional activation, attempts were made to discover genes, by subtractive hybridization of a cDNA library or by differential display, whose expression depended on the presence of BA; however these were unsuccessful. BA modulated the expression of a limited number of genes only. The possible mechanism by which BA could be promoting DNA metabolism during maize germination is discussed.

We have studied by means of cytology and autoradiography the effect of benzyladenine (BA, a synthetic cytokinin) on DNA metabolism during early maize germination.The data indicate that BA stimulates nuclear DNA replication. The doubling of the amount of nuclear DNA in BA-treated axes occurs earlier than in nontreated axes, and there is a three-fold increase in the mitotic index at 24 h of germination. These results provide further corroboration for the suggestion that the stimulation of DNA synthesis observed relates to a nuclear replicative type of synthesis. Possible mechanisms of action of BA are discussed.

The search for germination-specific genes has been a laborious and unrewarding task, since many of the genes expressed during germination are also expressed in embryogenesis or in other developmental stages. By using mRNA differential display of transcript populations from maize (Zea mays L.) embryo axes, germinated for different times with or without a previous osmopriming treatment, a 682 bp cDNA was isolated that was present only after 24 h germination, and absent during osmopriming or during early germination. Screening of a cDNA library using the 682 bp probe yielded a 1554 bp cDNA that contained an open reading frame coding for 436 amino acids. This gene, referred to as ZmAA9-24, was expressed in root tissues, but was not detected in shoot or leaf tissues. Expression of ZmAA9-24 occurred earlier during germination (by 15 h) if embryo axes were imbibed in the presence of cytokinins or if seeds were previously osmoprimed. The predicted protein sequence of ZmAA9-24 is 39.6% identical to the product of the recently identified Arabidopsis gene COBRA (54.5% in the central region), which appears to participate in the regulation of cell expansion, particularly in roots, and belongs to the glycosylphosphatidylinositol (GPI)-anchored protein family. ZmAA9-24 expression might be regulated by both cell expansion and the cell cycle, processes that have a central role during seed germination.