Internode Length in Pisum. A New Allele at the le Locus

In Pisum sativum L. a third, more severe, allele at the internodelength locus le is identified and named le^d. Plants homozygousfor le^d possess shorter internodes and appear relatively lessresponsive to GA₂₀ than comparable le (dwarf) plants. Gene le^dmay act by reducing the 3ß-hydroxylation of GA₂₀ tothe highly active GA₁ more effectively than does gene le. Theresults indicate that le is a leaky mutant and therefore thatendogenous GA₁ influences internode elongation in dwarf (le)plants. Pisum sativum, peas, internode length, genetics, gibberellin, dwarf elongation     

Internode Length in Pisum: the Response of le na Scions Grafted to Na Stocks

The recessive gene na results in peas with extremely short internodes(phenotype nana). In intact plants, na prevents expression ofthe Le/le gene pair which determine the tall/dwarf difference.na blocks a step early in the gibberellin biosynthetic pathwaywhile le prevents conversion of gibberellin A₂₀ to GA₁. Whengrafted to leafy Na stocks, le na and Le na scions become phenotypicallydwarf and tall, respectively. Hence, Na stocks provide a graft-transmissiblesubstance which promotes elongation of na scions and allowsexpression of the Le/le difference. Pisum, internode length, grafting, genotype     

Internode length and anatomical changes in Pisum genotypes crys and cryc in response to extended daylength and applied gibberellin A1

Dwarf pea (Pisum sativum L.) plants with genotypes cry^c and cry^s responded differently when an 8 h photoperiod (8 h daylight, 16 h dark) was extended to 24 h (8 h daylight, 16 h incandescent light). Genotype cry^c showed up to a 4-fold increase in internode length, sustained by increases in both cell length (particularly of epidermal cells) and cell number (particularly of cortical cells) while cry^s plants showed up to a 2-fold increase in internode length sustained mostly by an increase in cell number. Under an 8 h (daylight) photoperiod the two genotypes did not differ in their sensitivity to applied gibberellin A₁ (GA₁) and they showed a similar pattern of response. GA₁ significantly increased internode length, cell length and cell number in both genotypes. Incandescent light did not increase the size of the response to GA₁ except for cry^s plants at high dose rates of GA₁ (29–58 nmol). At saturating doses of GA₁ the two genotypes attained a similar peak internode length; incandescent light increased the peak by about 40%. GA₁ increased the rate of leaf appearance by up to 33% while incandescent light reduced the rate by 4–7%. The elongation response of the more mature internodes of cry^c plants to GA₁ or incandescent light was due primarily to an increase in cell length whereas increased cell number made a significant contribution in the case of internodes which were relatively immature at the time the stimulus was applied. The progressive increase in internode length of both genotypes during ontogeny was due primarily to an increase in cell number. In conclusion, alleles cry^c and cry^s (background le La) do not confer a difference in sensitivity to GA₁ and the increase in internode length in response to incandescent light is probably not the result of a real or perceived increase in GA₁ level. Allele cry^s may partially block a phytochrome mediated response to light and the key difference between genotypes cry^s and cry^c may lie in the greater elongation (extensibility?) of cry^c epidermal cells in incandescent light.     

Metabolism of Gibberellins in Cell-Free Extracts of Anthers from Normal and Dwarf Rice

Cell-free extracts were prepared from anthers of normal anddwarf rice (Oryza sativa L.), and the metabolism of radioisotope-labeledgibberellins in the extracts was analyzed by HPLC and gas chromatography-massspectrometry (GC/MS). GA₁₂ was converted to GA₁₅ and GA₃₄ inthe extracts. GA₂₀ was converted to GA¹, GA₈ and GA₂₉, but GA₉was converted only to GA₃₄. The extracts of the dwarf cultivar,Waito-C (dy mutant), showed the same 3ß-hydroxylationactivity as did those of the normal cultivar, Nihonbare, indicatingthat the dy gene is not expressed in the anthers. These resultssuggest that the regulation of the biosynthesis of gibberellinsin rice is organ-specific. (Received November 9, 1989; Accepted January 10, 1990)     

Influence of Gibberellic Acid and the Rht3 Gene on Choline and Phospholipid Metabolism in Wheat Aleurone Tissue

The effect of gibberellic acid (GA3) on phospholipid metabolismand -amylase production was studied in aleurone tissue of twonear-isogenic lines of wheat (Triticum aesuvum L.). Incubationof embryoectomized seeds from a GA-responsive line (rht3, tall)with GA₃ caused the induction of -amylase activity after a lagphase of 30 h. In the case of embryoectomized seeds from a ‘GA-insensitive’line (Rh13, dwarf), however, the lag phase was extended up to50 h. During the first 14 h following imbibition, GA₃ inhibitedcholine uptake and its subsequent incorporation into phosphatidylcholine in the Rhr3 line but not in the rht3 line. GA₃ promotedphospholipid breakdown in both the lines during this period,however. GA₃ also terminated independent turnover of the cholineN-methyl groups in phosphatidyl choline and promoted turnoverof the whole choline headgroup. These results are discussedin relation to the possibility that phosphatidyl choline turnoveris an integral part of the GA₃ signal-transduction mechanismin aleurone tissue. Key words: GA3, Rht3 gene, choline, phospholipid     

Internode length in pisum: do the internode length genes effect growth in dark-grown plants?

Internode length in light-grown peas (Pisum sativum L.) is controlled by the interaction of genes occupying at least five major loci, Le, La, Cry, Na, and Lm. The present work shows that the genes at all of the loci examined (Le, Cry, and Na) also exert an effect on internode length in plants grown in complete darkness. Preliminary results using pure lines were verified using either segregating progenies or near isogenic lines. The major cause of the differences was due to a change in the number of cells per internode rather than to an alteration of the cell length. Since the genes occupying at least two of these loci, Le and Na, have been shown to be directly involved with gibberellin metabolism, it appears that gibberellins are not only essential for elongation in the dark but are limiting for elongation in the nana (extremely short, na), dwarf (Na le), and tall (Na Le) phenotypes. These results are supported by the large inhibitory effects of AMO 1618 treatments on stem elongation in dwarf and tall lines grown in the dark and the fact that applied gibberellic acid could overcome this inhibition and greatly promote elongation in a gibberellin-deficient na line. It is clear that the internode length genes, and in particular the alleles at the Le locus, are not acting by simply controlling the sensitivity of the plant to light.     

Biological Activities of the Methyl Ester of Gibberellin A73, a Novel and Principal Antheridiogen in Lygodium japonicum

The synthetic methyl ester of GA₇₃ (GA₇₃-Me) and the naturalantheridiogen of Lygodium japonicum showed almost the same activityto induce the formation of antheridia in dark-grown protonemataof L. japonicum at concentrations of 10^-14 M and higher. Thus,it appears that the principal antheridiogen in L. japonicumis GA₇₃-Me. GA₇₃-Me inhibited formation of ar-chegonia in light-grownprothallia of L. japonicum at concentrations of 10^-11 M andhigher and induced germination of spores in the dark in thisspecies at the same range of concentrations. GA₇₃(free acidform) promoted growth of seedlings of dwarf rice and hypocotylsof cucumber seedlings at dosages of and above 1 and 100 ng/plant,respectively. Eight compounds related to GA₇₃-Me, includingantheridiogens of Anemia phyllitidis and Anemia mexicana, wereactive in inducing an-theridial formation in L. japonicum, althoughtheir activities were considerably lower than that of GA₇₃-Me. (Received August 24, 1988; Accepted November 28, 1988)     

Effects of Gibberellin on the Arrangement and the Cold Stability of Cortical Microtubules in Epidermal Cells of Pea Internodes

Longitudinal microtubules are predominant in epidermal cellsof the 3rd internodes of dwarf pea (Pisum sativum L. cv. LittleMarvel) seedlings. In more than 50% of the cells, cortical microtubulesare running parallel to the cell axis. GA₃ promotes elongation of the internodes and gives rise toa predominance of transverse microtubules. In more than 60%of the GA₃-treatd cells, cortical microtubules are running transverseto the cell axis. Longitudinal microtubules in the GA₃-untreated cells are resistantto low-temperature treatment, but transverse microtubules inthe GA₃-treated cells are sensitive to it. Longitudinal microtubulesare present in GA₃-treated epidermal cells with low frequency.They are resistant to low-temperature treatment. Longitudinal, oblique and transverse microtubules are presentwith almost the same frequency in epidermal cells of the 3rdinternodes of tall pea (cv. Early Alaska) seedlings. GA₃ promoteselongation of the internodes also in tall pea seedlings, butit does not alter the direction of cortical microtubules sodistinctly as it does in dwarf pea seedlings. As in dwarf pea seedlings, longitudinal microtubules are resistantto low-temperature treatment, and transverse microtubules aresensitive to it in tall pea seedlings. (Received September 19, 1986; Accepted December 26, 1986)     

THE INDEPENDENT ACTION OF MORPHACTINS AND GIBBERELLIC ACID ON HIGHER PLANTS

The comparative activity of three morphactins, n-butyl-9-hydroxyfluorene-9-carboxylate (IT 3233), 9-hydroxy-fluorene-9-carboxylicacid (IT 3235) and methyl-2-chloro-9-hydroxy fluorene-9-carboxylate(IT 3456) on the action of gibberellic acid (GA₃) were examinedutilizing dwarf pea (Progress No. 9), dwarf corn (strain d-5),CCC dwarfed Alaska pea, and embryoless barley half-seeds. When applied either prior to or simultaneously with GA₃ themorphactins were without effect in reducing the response ofthe dwarf peas to the gibberellin. In fact IT 3233 and IT 3235in combination with GA₃ produced taller plants than the GA₃controls. Similar results were observed using CCC dwarfed Alaskapeas. The morphactins did, however, produce several morphologicaleffects on the peas. In addition, they were effective in breakingapical dominance in the peas. The morphactins were also withouteffect on the dwarf corn bioassay and no morphological changeswere observed. In the embryoless barley half-seeds the morphactins did notinhibit the response of the tissue to the GA₃. Also, the morphactinsdid not exhibit gibberellin-like properties. The results suggest that morphactins do not compete with gibberellinfor similar sites of action. ¹Published with the approval of the Director of the MichiganAgricultural Experiment Station as Journal Article Number 3917.     

Endogenous Gibberellins in the Vegetative Shoots of Tall and Dwarf Cultivars of Phaseolus vulgaris L.

A series of 13-hydroxygibberellins, gibberellin A₁ (GA₁), GA₁₉,GA₂₀, GA₄₄ and GA₅₃, were identified by GC/MS (full scan) fromvegetative shoots of tall (cv. Kentucky Wonder) and dwarf (cv.Masterpiece) Phaseolus vulgaris L. It is suggested that GA₁is active per se in the control of shoot elongation of P. vulgarisL., and that dwarfism in Masterpiece is not due to shortageof the active GA, but to its low ability to respond to the bioactiveGA. (Received August 29, 1988; Accepted November 21, 1988)     

Ontogenetic variation in levels of gibberellin A1 in Pisum

The levels of the biologically active gibberellin (GA), GA₁, and of its precursor, GA₂₀, were monitored at several stages during ontogeny in the apical portions of isogenic tall (Le) and dwarf (le) peas (Pisum sativum L.) using deuterated internal standards and gas chromatography-selected ion monitoring. The levels of both GAs were relatively low on emergence and on impending apical arrest. At these early and late stages of development the internodes were substantially shorter than at intermediate stages, but were capable of large responses to applied GA₃. Tall plants generally contained 10–18 times more GA₁ and possessed internodes 2–3 times longer than dwarf plants. Further, dwarf plants contained 3–5 times more GA₂₀ than tall plants. No conclusive evidence for the presence of GA₃ or GA₅ could be obtained, even with the aid of [²H₂]GA₃ and [²H₂]GA₅ internal standards. If GA₃ and GA₅ were present in tall plants, their levels were less than 0.5% and 1.4% of the level of GA₁, respectively. Comparison of the effects of gene le on GA₁ levels and internode length with the effects of ontogeny on these variables shows that the ontogenetic variation in GA₁ content was sufficient to account for much of the observed variation in internode length within the wild-type. However, evidence was also obtained for substantial differences in the potential length of different internodes even when saturating levels of exogenous GA₃ were present.Abreviations GA_n gibberellin A_n We thank Noel Davies, Omar Hasan, Leigh Johnson, Katherine McPherson and Naomi Lawrence for technical help, Professor L. Mander (Australian National University, Canberra) for deuterated GA standards and the Australian Research Council for financial assistance.     

Internode length inPisum: biochemical expression of thele andna mutations in the slender phenotype

Thele andna mutations in pea block GA biosynthesis and normally cause a marked reduction in internode length. However, neither of these genes influences the growth of plants carrying thecry ^s la gene combination. Plants of this genotype have long, thin internodes, pale green foliage, and abnormal flower and fruit development, collectively referred to as the slender phenotype. [¹³C,³H]Gibberellin A₂₀ is metabolized to GA₁, GA₈, and GA₂₉ in slender lines carrying the geneLe but only to GA₂₉ and GA₂₉-catabolite inle lines. Examination of¹²C:¹³C isotopic ratios showed that metabolites were strongly diluted by endogenous [¹²C]GAs inNa lines. However, little if any significant dilution was observed in a line homozygous for thena gene. These results confirm that thele andna mutations are fully expressed at the biochemical level in slender phenotypes of peas and concur with previous reports that internode elongation is entirely independent of GA levels incry ^s la (slender) plants.     

The quantitative relationship between gibberellin A1 and internode growth in Pisum sativum L.

The metabolism and growth-promoting activity of gibberellin A₂₀ (GA₂₀) were compared in the internode-length genotypes of pea, na le and na Le. Gibberellin A₂₉ and GA₂₉-catabolite were the major metabolites of GA₂₀ in the genotype na le. However, low levels of GA₁, GA₈ and GA₈-catabolite were also identified as metabolites in this genotype, confirming that the le allele is a leaky mutation. Gibberellin A₂₀ was approximately 20 to 30 times as active in promoting internode growth of genotype na Le as of genotype na le. However, the levels of the 3-hydroxylated metabolite of GA₂₀, GA₈ (2-hydroxy GA₁), were similar for a given growth response in both genotypes. In each case a close linear relationship was observed between internode growth and the logarithm of GA₈ levels. A similar relationship was found on comparing GA₂₀ metabolism in the three genotypes le ^d, le and Le. The former mutation results in a more severe dwarf phenotype than the le allele (which has previously been shown to reduce the 3-hydroxylation of GA₂₀ to GA₁). These results indicate that GA₂₀ has negligible intrinsic activity and support the contention that GA₁ is the only GA active per se in promoting stem growth in pea.Abbreviations GA_n gibberellin A_n - GC-MS gas chromatography-mass spectrometry - HPLC high-pressure liquid chromatography     

The Distribution of Gibberellins in Vegetative Tissues of Pisum sativum L. : I. Biological and Biochemical Consequences of the le Mutation

The concentrations of endogenous gibberellin (GA) 1, 5, 8, 19, 20, and 29 in the component tissues of maturing tall (Le) and dwarf (le) pea (Pisum sativum) plants have been determined. The following conclusions were drawn from the data obtained: (a) GA₂₀ and its metabolites accumulate only in the growing regions of Le and le plants; (b) the le mutation is biochemically expressed in all immature tissues of the dwarf plants; (c) the quantitative composition of the GA metabolites in the various immature tissues is variable; (d) the total GA concentration in apical buds, unexpanded leaves, and tendrils is considerably higher than in GA₁-responsive stem tissue; and (e) there is very little GA accumulation of the inactive 2β-hydroxylated GAs (GA₈ and GA₂₉) in either the mature vegetative tissues or the roots of pea plants.     

Gibberellin A12 Is Converted to Gibberellin A53 in Cultured Cells of Nicotiana tabacum and Catharanthus roseus

The metabolism of GA₁₂ and its precursors was investigated incultured cells of seven cell lines of Nicotiana tabacum andthree cell lines of Catharanthus roseus using ^l4C-labeled substrates.The presence of a metabolic pathway from ent-7-hydroxykaurenoicacid to GA₅₃ via GA₁₂-aldehyde and GA₁₂ was demonstrated inthe cultured cells. GA₁₂ was effectively converted to GA₅₃ incells of BY-2, 2b-4, 2b-13 and CG from N. tabacum. By contrast,GA₅₃ was not converted to any other GAs in all of the linesof cells examined. The metabolism of C₁₉-GAs was also examinedusing ³H-labeled substrates. The conversion of GA₂₀ to GA₂₉and GA, and of GA₄ to GA₃₄ occurred more efficiently in cellsfrom C. roseus than in cells from N. tabacum. However, 13-hydroxylationof GA₄ and GA₉ was not observed in any of the cell culturesexamined. Among the various metabolites, GA₅₃, GA₂₉ and GA₃₄were identified by full-scan GC/MS. (Received December 20, 1990; Accepted May 27, 1991)     

The Interaction of Genetic, Environmental, and Hormonal Factors in Stem Elongation and Floral Development of Cucumber Plants

Hypocotyl length was found to vary between cucumber plants carryingdifferent genes controlling sex expression. Among lines havingonly unisexual flowers (genotype M/M), the homozygous monoeciousplants (st⁺/st⁺) had significantly longer hypocotyls than theirgynoecious counterparts (st/st), heterozygous gynoecious plants(st+/st) being intermediate. Similarly, hypocotyls of plantsof an andromonoecious line (st⁺/st⁺ m/m) were significantlylonger than in their hermaphrodite counterparts (st/st m/m).Differences in intemode length were also significant and inthe same direction. Since stem and particularly hypocotyl elongationin cucumber is known to be very sensitive to applied gibberellin,these findings suggest the existence of differences in the effectivelevels of endogenous gibberellins in the different sex types,higher levels being correlated with stronger male tendency.This conclusion is in good agreement with the known effect ofapplied gibberellin on sex expression (enhancement of the maletendency) in cucumber. Application of gibberellin (GA₄+GA₇) and exposure to ‘summer’conditions (long days and relatively high temperature) inhibitedthe development of pistillate flowers while ‘winter’conditions (short days, lower temperature) had a similar effecton staminate flowers. The effect was in either case specific,that is, limited to flower development. It is concluded thatexogenous and endogenous gibberellins affect not only the initiation,but also the further development, of flowers in the cucumber.     

Inoculation with Azospirillum lipoferum Affects Growth and Gibberellin Status of Corn Seedling Roots

Maize (Zea mays L., hybrid Cargill 147) seedlings, grown inaseptic conditions, were inoculated with three strains of Azospirillumlipoferum (Al op 33, Al iaa 320, and ATCC 29708) or culturedin different concentrations of indol-3-acetic acid (IAA) orgibberellin A₃ (GA₃). After 48 h, root length, root surfacearea, root dry weight, and shoot dry weight were measured inall treatments. Gibberellin content was evaluated in selectedroots of control and Azospirillum inoculated seedlings by gaschromatography-mass spectrometry-selected ion monitoring withthe use of deuterated gibberellins as internal standards. Thethree strains of A. lipoferum, IAA (2 ng ml^–1), and GA₃(40 to 400 pg ml^–1) significantly enhanced root growth.Improvement of root hair growth and density was obtained mainlywith A. lipoferum strain Al op 33 and GA₃ 40 pg ml^–1.GA₃ was identified by gas chromatography-mass spectrometry (byboth, full scan and selected ion monitoring) in a free acidfraction from roots of the seedlings inoculated with A. lipoferum.In the roots of the non inoculated seedlings GA₃ was found afterhydrolysis of a fraction expected to contain glucosyl conjugates. (Received April 26, 1993; Accepted September 27, 1993)     

A Crucial Role for Gibberellins in Stress Protection of Plants

The hypothesis that there is an intimate relationship betweengibberellin levels and plant stress protection has been testedusing near-isogenic lines of a normal and dwarf barley (Hordeumvulgare L.). Application of paclobutrazol (a triazole), inducedstress protection in the normal line and application of GA₃to the dwarf reversed the inherent stress tolerance. Reversalof the dwarf phenotype by specific gibberellins (GAs) suggeststhat the conversion of GA₂₀ to GA₁ and GA₉ to GA₄ has been compromised.These observations indicate that modulation of specific GAsplays a key role in stress protection. The preferential useof non-chemical technologies including enzyme regulation, phytochromeA overexpression, and breeding for induction of stress tolerancein plants are discussed. (Received November 26, 1998; Accepted March 11, 1999)     

Abscisic Acid and Apical Dominance in Phaseolus coccineus L. The Role of Tissue Age

Abscisic acid (ABA) applied to the decapitated second internodeof runner bean plants enhanced outgrowth of lateral buds onlywhen internode stumps were no longer elongating. Applied toelongating internodes of slightly younger plants, ABA causesinhibition of bud outgrowth. Together with 10^–4 M indol-3-ylacetic acid (IAA), ABA stimulated internode elongation and interactedadditively in the inhibition of bud outgrowth. A mixture of10^–5 M ABA and 10^–6 M gibberellic acid (GA₃ ) causedsimilar effects on internode growth as IAA + ABA, but was mutuallyantagonistic in effect on growth of the lateral buds. Abscisic acid, apical dominance, gibberellic acid, indol-3yl acetic acid, Phaseolus coccineus, bean