Establishment of fast-growing callus and root cultures of Cephalotaxus harringtonia

Summary Callus cultures were established from Cephalotaxus harringtonia (Japanese plumyew) stem expiants cultured on Murashige and Skoog medium supplemented with 4.5 M 2,4-dichlorophenoxyacetic acid and 0.05 M 6-furfurylaminopurine. The inclusion of 4.9 M 6-(,-dimethylallylamino) purine as the sole hormone significantly increased the growth rate of the callus. Organogenesis giving rise to both shoots and roots occurred upon transfer of the callus onto a hormonefree medium. Vitrification was common on all regenerated shoots cultured on Gelrite-containing medium. Regenerated roots were excised and established in McCown's woody plant medium. Doubling the phosphate and nitrate levels in the medium increased the growth of these root cultures.Abbreviations MS Murashige and Skoog basal medium - B5 Gamborg's B5 basal salt medium - WP McCown's woody plant basal salt medium - 2,4-D 2,4-dichlorophenoxyacetic acid - Kinetin 6-furfurylamino-purine - 2iP 6-(,-dimethylallylamino) purine - IBA indole-3-butyric acid     

Identification and characterization of cDNAs encoding ethylene biosynthetic enzymes from Pelargonium x hortorum cv Snow Mass leaves.

Two Pelargonium 1-aminocyclopropane-1-carboxylate (ACC) synthase cDNAs (GAC-1 and GAC-2) were identified and characterized. GAC-1 is 1934 bp long with a 1446-bp open reading frame encoding a 54.1-kD polypeptide. GAC-2 is a 1170-bp-long ACC synthase polymerase chain reaction fragment encoding 390 amino acids. Expression of GAC-1 and GAC-2 together with a previously identified ACC oxidase (GEFE-1) was examined in different Pelargonium plant parts, and leaves were subjected to osmotic stress (sorbitol), metal ion stress (CuCl2), auxin (2,4-dichlorophenoxyacetic acid [2,4-D]), and ethylene. GAC-1 expression was not detectable in any of the plant parts tested, whereas high levels of GAC-2 were expressed in the leaf bud, young leaf, young floret, fully open floret, and senescing floret. GAC-2 was expressed to a lesser degree in fully expanded leaves or roots and was undetectable in old leaves and floret buds. GEFE-1 was detectable at all leaf ages tested, in young and fully open florets, and in the roots; however, the highest degree of expression was in the senescing florets. GAC-1 was induced by sorbitol. Both GAC-1 and GAC-2 were only slightly affected by CuCl2 and induced indirectly by 2,4-D. GEFE-1 was highly induced by sorbitol, CuCl2, and 2,4-D. GAC-1, GAC-2, and GEFE-1 were unaffected by ethylene treatment. These results suggest that GAC-1 is only induced by stress and that GAC-2 may be developmentally regulated, whereas GEFE-1 is influenced by both stress and development.     

Brassinosteroid-induced exaggerated growth in hydroponically grown Arabidopsis plants

The effects of root application of brassinolide (BL) on the growth and development of Arabidopsis plants ( Arabidopsis thaliana ecotype Columbia [L.] Heynh) were evaluated. Initially, all leaves were evaluated on plants 18, 22, 26 and 29 days old. The younger leaves were found to exhibit maximal petiole elongation and upward leaf bending in response to BL treatment. Therefore, based on these results leaves 6, 7 and 8 on 22–24-day-old plants were selected for all subsequent studies. Elongation along the length of the petiole in response to BL treatment was uniform with the exception of an approximately 4 mm region next to the leaf where upward curvature was observed. Both BL and 24-epibrassinolide (24-epiBL) were evaluated, with BL being more effective at lower concentrations than 24-epiBL. The exaggerated growth induced by 0.1 μ M BL was not observed in plants treated with 1 000-fold higher concentrations of GA₃, IAA, NAA or 2,4-D (100 μ M ). In addition, no exaggerated growth effects were observed when plants were treated with 200 ppm ethylene or 1 m M ACC. All treatments with BL, NAA, 2,4-D, IAA or ACC promoted ethylene and ACC production in wild type Arabidopsis plants, but only BL triggered exaggerated plant growth. BL also promoted exaggerated growth and elevated levels of ACC and ethylene in the ethylene insensitive mutant etr1-3 , showing that the effect of BR on growth is independent of ethylene. This work provides evidence that BR-induced exaggerated growth of Arabidopsis plants is independent of gibberellins, auxins and ethylene.     

The Physiological,Biochemical and Molecular Roles of Brassinosteroids and Salicylic Acid in Plant Processes and Salt Tolerance

Plant hormones regulate plant growth and development by affecting an array of cellular, physiological, and developmental processes, including, but not limited to, cell division and elongation, stomatal regulation, photosynthesis, transpiration, ion uptake and transport, initiation of leaf, flower and fruit development, and senescence. Environmental factors such as salinity, drought, and extreme temperatures may cause a reduction in plant growth and productivity by altering the endogenous levels of plant hormones, sensitivity to plant hormones, and/or signaling pathways. Molecular and physiological studies have determined that plant hormones and abiotic stresses have interactive effects on a number of basic biochemical and physiological processes, leading to reduced plant growth and development. Various strategies have been considered or employed to maximize plant growth and productivity under environmental stresses such as salt-stress. A fundamental approach is to develop salt-tolerant plants through genetic means. Breeding for salt tolerance, however, is a long-term endeavor with its own complexities and inherent difficulties. The success of this approach depends, among others, on the availability of genetic sources of tolerance and reliable screening techniques, identification and successful transfer of genetic components of tolerance to desired genetic backgrounds, and development of elite breeding lines and cultivars with salt tolerance and other desirable agricultural characteristics. Such extensive processes have delayed development of successful salt-tolerant cultivars in most crop species. An alternative and technically simpler approach is to induce salt tolerance through exogenous application of certain plant growth–regulating compounds. This approach has gained significant interest during the past decade, when a wealth of new knowledge has become available on the beneficial roles of the six classes of plant hormones (auxins, gibberellins, cytokinins, abscisic acid, ethylene, and brassinosteroids) as well as several other plant growth–regulating substances (jasmonates, salicylates, polyamines, triacontanol, ascorbic acid, and tocopherols) on plant stress tolerance. Among these, brassinosteroids (BRs) and salicylic acid (SA) have been studied most extensively. Both BRs and SA are ubiquitous in the plant kingdom, affecting plant growth and development in many different ways, and are known to improve plant stress tolerance. In this article, we review and discuss the current knowledge and possible applications of BRs and SA that could be used to mitigate the harmful effects of salt-stress in plants. We also discuss the roles of exogenous applications of BRs and SA in the regulation of various biochemical and physiological processes leading to improved salt tolerance in plants.     

Inhibition of brassinosteroid-induced epinasty in tomato plants by aminooxyacetic acid and Co2+

The inhibitory effects of aminooxyacetic acid (AOA) and cobalt chloride (CoCl₂) on brassinosteroid (BR)-induced epinasty in tomato plants ( Lycopersicon esculentum Mill. cv. Heinz 1350) are evaluated. CoCl₂ dramatically decreases petiole bending and ethylene production as the concentration increases from 50 to 200 μ M. The content of 1-aminocyclopropane-1-carboxylic acid (ACC) in the petiole, instead of accumulating, is reduced and does not change over the concentration range tested. Inhibition of BR-induced epinasty by AOA results from inhibition of ACC synthesis. There are dramatic reductions in petiole bending, ethylene and ACC production as the concentration of AOA is increased from 50 to 200 μ M. Maximum inhibition occurs when the plants are pretreated with the inhibitors. The degree of inhibition increases as the length of pretreatment increases from 1 to 4 h. The response of BR-treated plants to AOA and CoCl₂ is similar to the effect of auxin, indicating the integral relationship between BR and auxin.     

Calcium-dependent protein kinase gene expression in response to physical and chemical stimuli in mungbean (Vigna radiata)

Protein kinases are important in eukaryotic signal transduction pathways. In this study we designed degenerate oligonucleotides corresponding to two conserved regions of protein kinases and using the polymerase chain reaction (PCR) have amplified a 141 bp fragment of DNA from mungbeans (Vigna radiata Rwilcz cv. Berken). Sequence analysis of the PCR products indicates that they encode several putative protein kinases with respect to their identity with other known plant protein kinases. Using one of the six fragments (CPK3-8), we isolated a 2022 bp cDNA (VrCDPK-1) from a Vigna radiata gt11 library. VrCDPK-1 has a 96 bp 5-untranslated region and a 465 bp 3-untranslated region and an open reading frame of 1461 bp. VrCDPK-1 contains all of the conserved regions commonly found in calcium dependent protein kinases (CDPK). VrCDPK-1 shares 24 to 89% sequence identity with previously reported sequences for plant CDPKs at the protein level. southern analysis revealed the presence of several copies of the CDPK gene. VrCDPK-1 expression was stimulated when mungbean cuttings were treated with CaCl₂, while treatment with MgCl₂ had no effect. We are reporting for the first time a CDPK gene in mungbean which is inducible by mechanical strain. Cuttings treated with indole-3-acetic acid (IAA) or subjected to salt stress showed an increase in VrCDPK-1 expression. There was a dramatic stimulation in VrCDPK-1 expression 6 h after cuttings were treated with cycloheximide.     

Increased photosynthetic rates following gibberellic acid treatments to the roots of tomato plants

This study was conducted to evaluate the effects of root applications of gibberellic acid (GA₃) on photosynthesis in tomato plants grown hydroponically. Photosynthetic rates (mg CO₂/dm²/hr) determined using an open infrared CO₂ gas exchange system showed a 40–50% increase within 5 hr after treatment with a 1.4 µM gibberellic acid (GA₃) to their roots. The effect was shown to persist for the duration of the experiment (9 days). Plants receiving pulses of 1.4 µM GA₃ to the roots for 1, 4, 8 or 12 hr exhibited significantly higher photosynthetic rates than the control for 6 days following treatment. By day 9 however, there was no significant difference. Continual treatments with 1.4 µM GA₃ to the roots maintained the photosynthetic rate significantly higher than the control for the duration of the experiment. Interestingly, at the lower light levels the percent stimulation was more dramatic. There was approximately a 90% increase in the photosynthetic rate at 80 µE m^-2 s^-1 while at saturating light conditions (560 µE m^-2 s^-1) there was approximately a 40% increase over the control rate. The light saturation point for both treated and control plants was 240 µE m^-2 s^-1. Applications of physiologically relevant concentrations of GA₃ to the roots of tomato plants stimulates photosynthesis more consistently than that achieved by previous studies involving foliar absorption.Approved for publication on May 28, 1981 as paper number 6242 in the Journal series of the Pennsylvania Agricultural Experiment Station.