zeatin-riboside and benzylaminopurine

Stay-green, a key trait of wheat, can not only increase the yield of wheat but also its resistance to heat stress during active photosynthesis. Cytokinins are the most potent general coordinator between the stay-green trait and senescence. The objectives of the present study were to identify and assess the effects of cytokinins on the photosynthetic organ and heat resistance in wheat. Two winter wheat cultivars, Wennong 6 (a stay-green cultivar) and Jimai 20 (a control cultivar), were subjected to heat stress treatment from 1 to 5 days after anthesis (DAA). The two cultivars were sprayed daily with 10 mg L-1 of 6-benzylaminopurine (6-BA) between 1 and 3 DAA under ambient and elevated temperature conditions. We found that the heat stress significantly decreased the number of kernels per spike and the grain yield (P < 0.05). Heat stress also decreased the zeatin riboside (ZR) content, but increased the gibberellin (GA3), indole-3-acetic acid (IAA), and abscisic acid (ABA) contents at 3 to 15 DAA. Application of 6-BA significantly (P < 0.05) increased the grain-filling rate, endosperm cell division rate, endosperm cell number, and 1,000-grain weight under heated condition. 6-BA application increased ZR and IAA contents at 3 to 28 DAA, but decreased GA3 and ABA contents. The contents of ZR, ABA, and IAA in kernels were positively and significantly correlated with the grain-filling rate (P < 0.05), whereas GA3 was counter-productive at 3 to 15 DAA. These results suggest that the decrease in grain yield under heat stress was due to a lower ZR content and a higher GA3 content compared to that at elevated temperature during the early development of the kernels, which resulted in less kernel number and lower grain-filling rate. The results also provide essential information for further utilization of the cytokinin substances in the cultivation of heat-resistant wheat.

Topics: Abscisic Acid; Benzyl Compounds; Cytokinins; Gibberellins; Hot Temperature; Indoleacetic Acids; Isopentenyladenosine; Plant Growth Regulators; Purines; Triticum

The cytokinin 6-benzylaminopurine (6-BAP) influences the embryogenic capacity of the tissues of Picea balfouriana during long subculture (after 3 months). Tissues that proliferate in 3.6 and 5 µM 6-BAP exhibit the highest and lowest embryogenic capacity, respectively, generating 113 ± 6 and 23 ± 3 mature embryos per 100 mg of tissue. In this study, a comparative transcriptomic and proteomic approach was applied to characterize the genes and proteins that are differentially expressed among tissues under the influence of different levels of 6-BAP. A total of 51 375 unigenes and 2617 proteins were obtained after quality filtering. There were 2770 transcripts for proteins found among these unigenes. Gene ontology (GO) analysis of the differentially expressed unigenes and proteins showed that they were involved in cell and binding activity and were enriched in ribosome and glutathione metabolism pathways. Ribosomal proteins, glutathione S-transferase proteins, germin-like proteins and calmodulin-independent protein kinases were up-regulated in the embryogenic tissues with the highest embryogenic ability (treated with 3.6 µM 6-BAP), which was validated via quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and these proteins might serve as molecular markers of embryogenic ability. Data are available via Sequence Read Archive (SRA) and ProteomeXchange with identifier SRP042246 and PXD001022, respectively.

Topics: Benzyl Compounds; Gene Expression Profiling; Indoleacetic Acids; Isopentenyladenosine; Kinetin; Molecular Sequence Annotation; Picea; Plant Somatic Embryogenesis Techniques; Proteome; Purines; Sequence Analysis, RNA; Transcriptome

The function of PsBRC1, the pea (Pisum sativum) homolog of the maize (Zea mays) TEOSINTE BRANCHED1 and the Arabidopsis (Arabidopsis thaliana) BRANCHED1 (AtBRC1) genes, was investigated. The pea Psbrc1 mutant displays an increased shoot-branching phenotype, is able to synthesize strigolactone (SL), and does not respond to SL application. The level of pleiotropy of the SL-deficient ramosus1 (rms1) mutant is higher than in the Psbrc1 mutant, rms1 exhibiting a relatively dwarf phenotype and more extensive branching at upper nodes. The PsBRC1 gene is mostly expressed in the axillary bud and is transcriptionally up-regulated by direct application of the synthetic SL GR24 and down-regulated by the cytokinin (CK) 6-benzylaminopurine. The results suggest that PsBRC1 may have a role in integrating SL and CK signals and that SLs act directly within the bud to regulate its outgrowth. However, the Psbrc1 mutant responds to 6-benzylaminopurine application and decapitation by increasing axillary bud length, implicating a PsBRC1-independent component of the CK response in sustained bud growth. In contrast to other SL-related mutants, the Psbrc1 mutation does not cause a decrease in the CK zeatin riboside in the xylem sap or a strong increase in RMS1 transcript levels, suggesting that the RMS2-dependent feedback is not activated in this mutant. Surprisingly, the double rms1 Psbrc1 mutant displays a strong increase in numbers of branches at cotyledonary nodes, whereas branching at upper nodes is not significantly higher than the branching in rms1. This phenotype indicates a localized regulation of branching at these nodes specific to pea.

Topics: Arabidopsis Proteins; Benzyl Compounds; Cytokinins; Gene Expression Regulation, Plant; Isopentenyladenosine; Kinetin; Lactones; Molecular Sequence Data; Mutation; Pisum sativum; Plant Growth Regulators; Plant Proteins; Plant Shoots; Purines; Signal Transduction; Transcription Factors; Up-Regulation; Xylem

When cytokinin-autonomous tobacco BY-2 cell cultures are transferred into 2,4-dichlorophenoxyacetic acid (2,4-D)-deprived medium, amyloplast development is initiated. Using this in vitro amyloplast-inducing system, the role of cytokinins in amyloplast formation was investigated. We show that addition of lovastatin, an inhibitor of mevalonate synthesis, to amyloplast-inducing medium reduced starch accumulation. Microscopic observation also revealed that lovastatin treatment decreased starch deposition; however, the overall morphologies of cells and plastids were less affected than control cell cultures. In addition, lovastatin lowered the transcription level of the ADP-glucose pyrophosphorylase small subunit (AgpS) gene. Application of mevalonate or zeatin dramatically restored the decrease in starch deposition, and restored AgpS mRNA accumulation. Moreover, addition of other molecules with cytokinin activity, such as adenine- and phenylurea-type compounds, restored starch accumulation and AgpS transcript levels, whereas other isopentenyl pyrophosphate-derived phytohormones did not. Liquid chromatography-mass spectrometry/mass spectrometry quantification of endogenous cytokinins revealed that endogenous cytokinins increased when BY-2 cells were transferred into 2,4-D-deprived medium from conventional medium containing 2,4-D. In addition, lovastatin treatment decreased endogenous cytokinins to some extent when cultured under 2,4-D-deprived conditions. Our results suggest that both 2,4-D deprivation and an increase in endogenous cytokinins have important roles in accelerating the changes in plastid morphology, starch accumulation, and AgpS gene expression.

Topics: 2,4-Dichlorophenoxyacetic Acid; Adenine; Adenosine; Benzyl Compounds; Cells, Cultured; Cytokinins; Gene Expression Regulation; Glucose-1-Phosphate Adenylyltransferase; Hemiterpenes; Isopentenyladenosine; Kinetin; Lovastatin; Mass Spectrometry; Mevalonic Acid; Nicotiana; Nucleotidyltransferases; Organophosphorus Compounds; Plant Growth Regulators; Plastids; Purines; RNA, Messenger; Starch; Zeatin