David M. Reid

Plant hormones and plant growth regulators in plant tissue culture

SummaryThis is a short review of the classical and new, natural and synthetic plant hormones and growth regulators (phytohormones) and highlights some of their uses in plant tissue culture. Plant hormones rarely act alone, and for most processes— at least those that are observed at the organ level—many of these regulators have interacted in order to produce the final effect. The following substances are discussed: (a) Classical plant hormones (auxins, cytokinins, gibberellins, abscisic acid, ethylene and growth regulatory substances with similar biological effects. New, naturally occurring substances in these categories are still being discovered. At the same time, novel structurally related compounds are constantly being synthesized. There are also many new but chemically unrelated compounds with similar hormone-like activity being produced. A better knowledge of the uptake, transport, metabolism, and mode of action of phytohormones and the appearance of chemicals that inhibit synthesis, transport, and action of the native plant hormones has increased our knowledge of the role of these hormones in growth and development. (b) More recently discovered natural growth substances that have phytohormonal-like regulatory roles (polyamines, oligosaccharins, salicylates, jasmonates, sterols, brassinosteroids, dehydrodiconiferyl alcohol glucosides, turgorins, systemin, unrelated natural stimulators and inhibitors), as well as myoinositol. Many of these growth active substances have not yet been examined in relation to growth and organized developmentin vitro.

Shifts in carbon isotope ratios of two C3 halophytes under natural and artificial conditions

SummaryThe total carbon δ13C values of two C3 halophytes,Salicornia europaea L. ssp.rubra (Nels.) Breitung andPuccinellia muttalliana (Schultes) Hitch., native to inland saline areas of Alberta, Canada, were determined for plants grown under controlled conditions of supplied NaCl in the nutrient solution, and for plants found growing in the field. Field specimens were collected along line transects which ran from areas of high salinity to areas of low salinity across the pattern of species zonation. The δ13C value of the two species seemed to reflect the water potential of the soil (ψwsoil) as measured arbitrarily at a depth of 10 cm, becoming less negative as the ψwsoil decreased. Over a linear distance of 5.55 m,S. europaea spp.rubra showed a shift of +5.3‰ as the ψwsoil went from-25x102 kPa to a minimum of-73x102 kPa. ForP. nuttalliana, the δ13C values differed by 3.4‰ over a distance of 7.45 m where the maximum difference in ψwsoil was 12.7x102 kPa. However, δ13C values ofP. nuttalliana only roughly reflected the spatial trends in ψwsoil at the time of collection. In the growth chamber, the δ13C value ofS. europaea ssp.rubra changed by a maximum of +8.0‰ when the solute potential of the nutrient solution (ψwsoil) was dropped from-0.25x102 kPa to-64.25x102 kPa; while the δ13C value ofP. nuttalliana changed by a maximum of +10.8‰ when the ψwsoil was dropped from-0.25x102 kPa to-40.25x102 kPa. Linear regression analyses indicated that the δ13C values of both species were strongly correlated (P<0.2%) with ψwsoil. The observed shifts in δ12C may represent changes in the mode of photosynthetic CO2 fixation. However, a number of other explanations, some of which are discussed in the text, are also possible. A proper ecophysiological interpretation of such shifts in δ13C values of C3 plants awaits a better understanding of the isotope fractionation mechanisms involved.

The effects of flooding on the export of gibberellins from the root to the shoot

SummaryGibberellin (GA)-like substances are shown to be present in the bleeding sap of tomatoes. It is demonstrated that these substances can promote stem elongation in intact tomato plants. Waterlogging of the root system inhibits stem growth and severely reduces the export of GAs from the root to the shoot in the xylem sap. It is suggested that decline in growth rate following waterlogging may, in part, be due to the reduction in the supply of GAs to the shoot.

Evolution of phenotypic plasticity in the Stellaria longipes complex : comparisons among cytotypes and habitats

Variation in the amount and pattern of plasticity was studied in three cytotypes (4x, 6x, and 8x) of Stellaria longipes and diploids of its suspected progenitor S. longifolia. All 13 traits considered showed plasticity. There were significant differences among cytotypes and habitats in plasticity for many traits. Overall, the diploids, S. longifolia, were most plastic, and the three cytotypes of S. longipes did not differ in amount of plasticity. Stellaria longifolia showed divergence from S. longipes in the pattern of plasticity as well. In general, cytotypes with more similar chromosome numbers had the same pattern of plasticity for more traits. Individuals from tundra populations differed in their pattern of plasticity from those of montane, boreal, and prairie origin, which were more similar to one another. Differences in plasticity among cytotypes were due primarily to divergence in amount, while differences among habitats were most often accounted for by divergent patterns of plasticity. We conclude that both polyploidy and natural selection have affected the evolution of plastic responses in this species complex. Analysis of the correlation between pairs of traits provided evidence that the pattern and amount of plasticity operate independently of one another and may be evolving separately.

gamma-Aminobutyric acid stimulates ethylene biosynthesis in sunflower.

[gamma]-Aminobutyric acid (GABA), a nonprotein amino acid, is often accumulated in plants following environmental stimuli that can also cause ethylene production. We have investigated the relationship between GABA and ethylene production in excised sunflower (Helianthus annuus L.) tissues. Exogenous GABA causes up to a 14-fold increase in the ethylene production rate after about 12 h. Cotyledons fed with [14C]GABA did not release substantial amounts of radioactive ethylene despite its chemical similarity to 1-aminocyclopropane-1-carboxylic acid (ACC), indicating that GABA is not likely to be an alternative precursor for ethylene. GABA causes increases in ACC synthase mRNA accumulation, ACC levels, ACC oxidase mRNA levels, and in vitro ACC oxidase activity. In the presence of aminoethoxyvinylglycine or [alpha]-aminoisobutyric acid, GABA did not stimulate ethylene production. We therefore conclude that GABA stimulates ethylene biosynthesis mainly by promoting ACC synthase transcript abundance. Possible roles of GABA as a signal transducer are suggested.

Ethylene, gibberellins, auxin and the apical control of branch angle in a conifer, Cupressus arizonica

Decapitation, gibberellin A3, high light, their combination, and certain levels of indole-3-acetic acid increase ethylene evolution and also induce branch hyponasty (upturning) in seedlings of Cupressus arizonica Greene, the increase in ethylene preceding obvious hyponasty. Exogenous ethylene also causes branch hyponasty and branches of seedlings maintained in an atmosphere scavenged of ethylene by mercuric perchlorate grow downwards. It is concluded that ethylene may play a role in the apical control of branch angle in some conifers. The positive effect of ethylene in increasing branch hyponasty may be direct, or reflect changes in levels of endogenous auxin and/or gibberellin.

The effect of drought on levels of abscisic acid, cytokinins, gibberellins and ethylene in aeroponically-grown sunflower plants

Abscisic acid (ABA), cytokinins and gibberellin-like substances (GAs) were extracted from the roots and shoots of 17-day-old sunflower seedlings which had been droughted or were unstressed. Plants were grown in an aeroponic chamber which allowed for good control over degree of water stress and easy access to roots. Following methanolic extraction of lyophilized material, cytokinins were separated from the acidic growth-regulators on a cellulose PO4 cationic exchange column. The cytokinins were analysed by paper chromatography and HPLC and the soybean hypocotyl section assay. Semipurified acidic regulators were chromatographed on SiO2 columns and HPLC and aliquots assayed with the dwarf rice cv. Tan-ginbozu bioassay for GAs. Fractions known to contain ABA were purified by sequential reverse-phase HPLC of the acid and then of the methyl ester forms followed by quantitation as Me-ABA on GLC-EC. ABA losses were measured by using an internal standard [3H]-ABA). Ethylene production was also monitored in stressed and unstressed seedlings.The effect of drought on GAs and ethylene was minimal. The ABA levels were markedly higher in droughted plants. Stressed roots had 32 times more ABA than controls. The levels of cytokinins in the shoots of droughted plants were about half those in unstressed shoots, and qualitative differences occurred in the roots. Under stress a large peak of activity was present similar to zeatin glucoside which was not present in the unstressed condition. The results are discussed in relation to drought-effects on metabolism.

Effects of Benzyladenine on the Growth of Waterlogged Tomato Plants

SummaryTreatment of waterlogged tomato plants with benzyladenine relieved most of the symptoms of flooding injury. The effects of benzyladenine included maintenance of chlorophyll levels, reduced epinastic curvature of petioles, absence of adventitious roots, and stem growth greater than that of waterlogged control plants.

CCC-Induced increase of gibberellin levels in pea seedlings

SummaryPea seedlings (cv. Alaska), were treated with two concentrations of (2-chloroethyl)trimethylammonium chloride (CCC) and choline chloride. Treatment with 1 mg/l CCC resulted in as much as a 150fold increase in endogenous gibberellin (GA) levels without there being any parallel stimulation of growth. Plants grown in 1,000 mg/l CCC were severely dwarfed but contained GA levels not significantly different from control plants grown in distilled water. CCC also retarded GA3-induced growth of pea seedlings. These effects appear to be CCC specific as the CCC analogue choline chloride affected neither the GA content of pea seedlings nor their response to GA3. The lack of correlation between endogenous GA levels and stem height suggests that in peas the predominant factor in CCC-induced inhibition of stem growth is not related to an effect of CCC on GA biosynthesis.

Pea PR 10.1 is a ribonuclease and its transgenic expression elevates cytokinin levels

The constitutive expression of a cDNA encoding a pea (Pisum sativum L.) PR 10 protein in Brassica napus leading to an enhancement of germination under saline conditions has been previously reported. In order to understand the biochemical function of this pea PR 10 protein, its cDNA has been expressed in Escherichia coli and the recombinant protein purified to homogeneity. Ribonuclease activity of the recombinant pea PR 10 protein has been demonstrated for the first time using an in-solution as well as an in-gel RNA degradation assay. Furthermore, in order to characterize the changes brought about as a result of the constitutive expression of the pea PR 10 cDNA in B. napus, we have measured the endogenous concentrations of several phytohormones. Increased cytokinin and, decreased abscisic acid (ABA) were observed in 7-day-old transgenic seedlings whereas no significant changes in the concentrations of gibberellin (GA) or indoleacetic acid (IAA) were observed at this stage of growth and development. The potential role(s) of PR 10 proteins with RNase activity and elevated cytokinins during plant stress responses as well as the possible relationship between PR 10 protein and changes in cytokinin concentrations are discussed.