Reversible conjugation of gibberellins in situ in maize

Gibberellins [³H]GA₄ (1.33 Curies per millimole) and [³H]GA₂₀ (2.36 Curies per millimole) were injected into the shanks of maize (Zea mays L.) cobs during rapid grain filling and mature seeds were subsequently harvested. Extracts of mature, dry seeds from 1980 feeds yielded only 20 to 30% of the ³H radioactivity in acidic, ethyl acetate-soluble form, and this was principally associated with the precursor, with lesser amounts of the major metabolite, [³H]GA₁ (putative identification based on sequential SiO₂ partition, and gradient-eluted reverse-phase C₁₈ high performance liquid chromatography [HPLC]). Most of the radioactivity in the dry seeds was associated with compounds having partition characteristics of, and co-chromatographing on, sequential SiO₂ partition and reverse-phase HPLC with glucosyl conjugates of the precursors (GA₄ or GA₂₀) and their probable major metabolite (GA₁). The majority of conjugate associated with the precursor GA₄ eluted coincidental with GA₄ glucoside. Subsequent acid or enzymic hydrolysis (β-glucosidase or cellulase) yielded the free GAs, putative identification being based on isocratic HPLC of each ³H-labeled conjugate → hydrolysis → isocratic HPLC of the ³H-labeled hydrolysate. Upon imbibition of the seeds, radioactivity associated with the conjugate fraction decreased; concomitantly, statistically significant increases in levels of free [³H]GA-like compounds were observed. Although the specific ratios of GA-like and GA-glucosyl conjugate-like substances varied substantially across years, hybrids, and even, in different plants from the same hybrid, this `reversible conjugation' (i.e. apparent conjugation during seed maturation followed by release of the GA moiety during germination), was reproducible for [³H]GA₂₀ in seed from two maize hybrids produced over 2 years.     

Relationships between seed germination requirements and ecophysiological characteristics aid the recovery of threatened native plant species in Western Australia

Summary One of the foremost technical issues addressed in reintroduction and restoration projects is the feasibility of establishing living plants. To advance the recovery process, the germination requirements of 201 threatened Western Australian seed‐bearing taxa representing a range of life forms, families and ecophysiological characteristics were studied. Procedures used to stimulate germination in otherwise dormant seed involved pretreatment using thermal shock, scarification, seed coat removal, soaking in an aqueous smoke solution and/or additions of the growth hormone gibberellic acid (GA₃). Sixty‐one taxa germinated under the basic trial conditions of light (12‐ h photoperiod), temperature (constant 15°C) and moisture, without additional pretreatments. These taxa were generally those with canopy‐stored seeds in the families Proteaceae and Casuarinaceae, and small‐seeded taxa in Myrtaceae. Taxa with soil‐stored seeds required single or multiple cues to stimulate germination. Seeds in the families Fabaceae and Mimosaceae were dependent on cracking of the seed coat, mechanically through nicking of the testa or through thermal shock, as were several non‐leguminous species of the Sterculiaceae and Liliaceae. Complete or partial removal of seed coats, in conjunction with GA₃ enhanced germination percentage in some taxa of the Myoporaceae, Lamiaceae and Myrtaceae. Application of GA₃ also enhanced germination percentage in members of the Epacridaceae. Several taxa previously stimulated by aqueous smoke solutions were equally responsive to additions of GA₃ after complete seed coat removal. In general, species with seed weights greater than 10 mg germinated better under a range of conditions than those with lighter seeds. There was no difference in germinability between resprouter and seeder species, and no obvious relationship between seed weight and germination rate. In the light of previous studies these results indicate that the relationship between germination requirements and ecophysiological characteristics is similar for both threatened and common species. These data will enable better prediction of likely dormancy breaking cues for other related species and will greatly assist the process of recovery and restoration work for mining operations and community bushland regeneration as well as single species reintroductions.     

Effects of smoke water and karrikin on seed germination of 13 species growing in China

Plant-derived smoke water (SW), derived from combusted plant material, has been shown to stimulate seed germination and improve seedling vigor of a number of plant species from fire-dependent Mediterranean-type climate areas. The effects of SW on seed germination of 13 plant species from southern tropical and subtropical monsoon climate regions of South China are reported for the first time in this study using laboratory and pot trials. Among the 13 species tested, only Aristolochia debilis showed a significant positive response to commercial SW when diluted 1:10. Seed germination of A. debilis was also stimulated by 1–100 nM 3-methyl-2H-furo [2, 3-c] pyran-2-one (karrikin 1 or KAR₁) and by 10–1000 µM gibberellic acid (GA₃). GA₃ stimulated seed germination of Santalum album and significantly elongated the radicles of A. debilis while SW could not. The functions and/or metabolic pathways of Kar₁ and GA₃ are likely to be separate and/or distinct.     

Factors Influencing Spore Germination and Early Gametophyte Development in Anemia mexicana and Anemia phyllitidis

Spores of Anemia mexicana Klotzsch and Anemia phyllitidis (L.) Swartz were tested comparatively to investigate the effects of various treatments on spore germination and early gametophyte development in light and darkness. The optimum pH for induction of spore germination is approximately 6. Both species have a minimum 8 hour light insensitive preinduction phase for spore germination. An additional 8 to 12 hours of light are needed to induce 50% germination in A. phyllitidis while at least 24 hours of light are needed for A. mexicana spores. A. phyllitidis has greater sensitivity to the four gibberellic acids tested (GA₃, GA₄, GA₇, and GA₁₃) than A. mexicana for induction of spore germination in darkness. In both species the greatest response was observed with GA₄ and GA₇. GA₁₃ was clearly the least effective. Gametophytes of each species are 100 times more sensitive to their own antheridiogen than to the antheridiogen of the other species. AMO-1618 (1 millimolar), fenarimol (1 mm), and ancymidol (0.1 mm) had essentially no effect on light-induced germination. The latter two did, however, inhibit gametophyte development.     

Dormancy and germination in Stachys germanica L. subsp. bithynica (Boiss.) Bhattacharjee seeds: Effects of short-time moist chilling and plant growth regulators

We investigated the germination requirements of the species Stachys germanica L. subsp. bithynica (Boiss.) Bhattacharjee (Lamiaceae). We studied the effects of scarification, short-time moist chilling (+4 °C) for 15 and 30 days, and various doses of gibberellic acid (GA₃; 0, 100, 150 and 250 ppm), Kinetin (KIN; 50 ppm) and a combination of 250 ppm GA₃ and 50 ppm KIN. The hormone and moist chilling treatments were carried out under both continuous darkness (20 °C) and photoperiodic (20/10 °C; 12/12 h, respectively) conditions. Seeds failed to germinate in response to short-time moist chilling treatments with distilled water under both continuous darkness and photoperiodic conditions. Seeds were found to have dormancy. Treatments with GA₃ or a combination of GA₃ and KIN were successful at breaking seed dormancy. A maximum of 37% of the seeds germinated after GA₃ application in all series. When only KIN was applied at a 50 ppm concentration, germination (12%) was found only with moist chilling for 30 days under continuous darkness. The highest germination rates were found in seeds treated with combination of 250 ppm GA₃ and 50 ppm KIN. In the combination treatments, while the moist chilling treatments for 15 days resulted in 68 and 73% germination, respectively, these rates were up to 95% in the moist chilling treatments for 30 days under continuous darkness and photoperiodic conditions. Mean germination time (MGT) in GA₃ and KIN combinations was lower than in other treatments. Scarification with 80% sulphuric acid did not promote germination. The characteristics of physiological dormancy of S. germanica ssp. bithynica seeds are consistent with conditions of existence in the in alpine habitat of this species.     

Identification of gibberellins from sugarcane plants

Five GAs, GA₁, GA₃, GA₁₉, GA₂₀, and GA₂₉, were identified in extracts from mature leaf and shoot apical meristem of flowering and non-flowering sugarcane (Saccharum spp. hybrids) by combined GC/MS. The presence of ABA was also confirmed.     

Impact of dormancy regulating chemicals on salinity induced dormancy in Lasiurus scindicus and Panicum turgidum: two desert glycophytic grasses

The indigenous forage grasses Lasiurus scindicus and Panicum turgidum are candidate species for the restoration of degraded desert rangelands. The impact of five dormancy regulating chemicals on overcoming salinity-induced germination inhibition was assessed under the best germination conditions in the two species. Seeds were germinated in a series of NaCl concentrations: 0–200 mM NaCl for P. turgidum, and 0–300 mM NaCl for L. scindicus. Lasiurus scindicus seeds were more tolerant to salinity than those of P. turgidum. Twenty percent of P. turgidum seeds germinated in 100 mM NaCl and none in the higher levels, but 47.5% and 8.8% of L. scindicus seeds germinated in 100 and 200 mM NaCl, respectively. The five studied chemicals (fusicoccin, GA₃, kinetin, nitrate and thiourea) did not succeed in improving germination of non-saline treated seeds of the two species, compared to the control, except thiourea in P. turgidum. The salinity-induced germination inhibition in P. turgidum was completely alleviated by the application of gibberellic acid (GA₃), partially alleviated by the application of fusicoccin, kinetin and thiourea, but not affected by nitrate. In L. scindicus, the germination inhibition was completely alleviated by fusicoccin, GA₃, nitrate and thiourea, but partially alleviated by kinetin. For using the two grass species in restoration of degraded rangelands affected by higher salinity, the results suggest using fusicoccin, GA₃, nitrate and thiourea with L. scindicus and GA₃ with P. turgidum seeds as a preseeding treatment can overcome the problem of reduced germination.     

Gibberellin concentration and transport in genetic lines of pea : effects of grafting

Effects of the Na and Le loci on gibberellin (GA) content and transport in pea (Pisum sativum L.) shoots were studied. GA₁, GA₈, GA₁₇, GA₁₉, GA₂₀, GA₂₉, GA₄₄, GA₈ catabolite, and GA₂₉ catabolite were identified by full-scan gas chromatography-mass spectrometry in extracts of expanding and fully expanded tissues of line C79-338 (Na Le). Quantification of GAs by gas chromatography-single-ion monitoring using deuterated internal standards in lines differing at the Na and Le alleles showed that na reduced the contents of GA₁₉, GA₂₀, and GA₂₉ on average to <3% and of GA₁ and GA₈ to <30% of those in corresponding Na lines. In expanding tissues from Na le lines, GA₁ and GA₈ concentrations were reduced to approximately 10 and 2%, respectively, and GA₂₉ content increased 2- to 3-fold compared with those in Na Le plants. There was a close correlation between stem length and the concentrations of GA₁ or GA₈ in shoot apices in all six genotypes investigated. In na/Na grafts, internode length and GA₁ concentration of nana scions were normalized, the GA₂₀ content increased slightly, but GA₁₉ levels were unaffected. Movement of labeled GAs applied to leaves on Na rootstocks indicated that GA₁₉ was transported poorly to apices of na scions compared with GA₂₀ and GA₁. Our evidence suggests that GA₂₀ is the major transported GA in peas.     

Gibberellins and parthenocarpic ability in developing ovaries of seedless mandarins

Satsuma (Citrus unshiu [Mak] Marc.) and Clementine (Citrus reticulata [Hort.] Ex. Tanaka, cv Oroval) are two species of seedless mandarins differing in their tendency to develop parthenocarpic fruits. Satsuma is a male-sterile cultivar that shows a high degree of natural parthenocarpy and a high fruit set. Seedless Clementine varieties are self-incompatible, and in the absence of cross-pollination show a very low ability to set fruit. The gibberellins (GAs) GA53, putative 17-OH-GA₅₃, GA₄₄, GA₁₇, GA₁₉, GA₂₀, GA₂₉, GA₁, 3-epi-GA₁, GA₈, GA₂₄, GA₉, and GA₄ have been identified from developing fruits of both species by full-scan combined gas chromatography-mass spectrometry. Using selected ion monitoring with [²H₂]- and [¹³C]-labeled internal standards, the levels of GA₅₃, GA₄₄, GA₁₉, GA₂₀, GA₁, GA₈, GA₄, and GA₉ were determined in developing ovaries at anthesis and 7 days before and after anthesis, from both species. Except for GA8, levels of the 13-hydroxy-GAs were higher in Satsuma than in Clementine, and these differences were more prominent for developing young fruits. At petal fall, Satsuma had, on a nanograms per gram dry weight basis, higher levels of GA₅₃ (10.4x), GA₄₄ (13.9x), GA₁₉ (3.0x), GA₂₀ (11.2x), and GA₁ (2.0x). By contrast, levels of GA₈ were always higher in Clementine, whereas levels of GA₄ did not differ greatly. Levels of GA₉ were very low in both species. At petal fall, fruitlets of Satsuma and Clementine contained 65 and 13 picograms of GA₁, respectively. At this time, the application of 25 micrograms of paclobutrazol to fruits increased fruit abscission in both varieties. This effect was reversed by the simultaneous applications of 1 microgram of GA₃. GA₃ alone improved the set in Clementine (13x), but had little influence on Satsuma. Thus, seedless fruits of the self-incompatible Clementine mandarin may not have adequate GA levels for fruit set. Collectively, these results suggest that endogenous GA content in developing ovaries is the limiting factor controlling the parthenocarpic development of the fruits.     

Quantitation of Gibberellins A(1), A(3), A(4), A(9) and a Putative A(9)-Conjugate in Grafts of Sitka Spruce (Picea sitchensis) during the Period of Shoot Elongation

The levels of endogenous gibberellin A₁ (GA₁), GA₃, GA₄, GA₉, and a cellulase hydrolyzable GA₉ conjugate in needles and shoot stems of mature grafts of Sitka spruce (Picea sitchensis [Bong.] Carr.) grown under environmental conditions that were either inductive, hot, and dry, or noninductive, cool, and wet, for flowering, were estimated by combined gas chromatography-mass spectrometry selected ion monitoring using deuterated [²H₂]GA₁, GA₃, GA₄, and GA₉ as internal standards. The samples were taken when the shoots had elongated about 30, 70, and 95% of the final shoot length and 17 days after elongation had terminated. The concentration of putative GA₉-conjugate, estimated by GCSIM of GA₉ after cellulase hydrolysis of the highly water soluble fraction, was 33 nanograms per gram fresh weight in the needles of both heat and drought- and cool and wet-treated plants sampled just after bud burst. The concentration gradually decreased to a final value of 13 nanograms per gram fresh weight in the heat and drought-treated grafts and 6 nanograms per gram fresh weight in the cool and wet-treated grafts. The stems contained no detectable putative GA₉ conjugate. Free GA₉ was highest in heat and drought-treated material. For plants subjected to this treatment, GA₉ increased from 22 to 32 nanograms per gram fresh weight in needles and from 1 to 22 nanograms per gram fresh weight in stems during the rapid stem elongation phase. By day 17, after cessation of shoot elongation, GA₉ had decreased to 12 nanograms per gram fresh weight in needles and 9 nanograms per gram fresh weight in the shoot stems. The cool and wet-treated material also showed an increase in GA₉ concentration during shoot elongation. However, the concentration was not as high and was also delayed compared with heat and drought-treated material. By day 17, after cessation of shoot elongation, GA₉ concentration was 9 nanograms per gram fresh weight in needles and 5 nanograms per gram fresh weight in stems for cool and wet treatment plants. The concentration of GA₄ was very low in tissue from both treatments. Fluctuation in concentration of the more polar gibberellins, GA₁ and GA₃, showed the same pattern as fluctuations in the content of GA₉. However, the heat and drought-treated material had lower amounts of GA₁ and GA₃ during the later phases of shoot elongation, than the cool and wet-treated material. These results imply differential metabolism between clones treated with conditions inductive and noninductive for flowering. Higher concentrations of putative GA₉ conjugate and free GA₉ in the hot and dry treatment indicate a higher capacity of synthesizing, for flowering, the physiologically important GA₄ in the heat and drought-treated material. This synthesis does not, however, result in a buildup of the GA₄ pool, probably because of a high turnover rate of GA₄. The cool and wet-treated material had higher amounts of GA₁ and GA₃, indicating that the differentiation was preferentially directed toward vegetative growth.     

The endogenous gibberellins of dwarf mutants of lettuce

The gibberellin (GA) content of E-1, a tall genotype of early flowering lettuce (Lactuca sativa L.), and of three selected GA-responsive dwarfs, dwf1, dwf2, and dwf2¹, has been determined using ¹³C-labeled internal standards and gas chromatographymass spectrometry (GC-MS). In the shoots of the E-1 parent, GA₁, 3-epi-GA₁, GA₃, GA₅, GA₈, GA₁₉, GA₂₀, GA₂₉, and GA₅₃ were identified by full scan GC-MS and Kovats retention indices. Purification by immunoaffinity chromatography selective for 13-hydroxy GAs, was necessary for GA identification. Relative to the parent E-1, the concentrations of GA₁, GA₈, GA₂₀, and GA₂₉ in the shoots of dwf2 plants were reduced to about 10% and in shoots of dwf2¹ plants to less than 50%. In dwf1 the levels of GA₁, GA₈, and GA₂₉ were also reduced to less than 50% of the parent E-1, but the level of GA₂₀ was fivefold higher than in E-1. Plant height was correlated with the endogenous levels of GA₁ and GA₈.     

Ethylene-Mediated Regulation of Gibberellin Content and Growth in Helianthus annuus L

Elongation of hypocotyls of sunflower can be promoted by gibberellins (GAs) and inhibited by ethylene. The role of these hormones in regulating elongation was investigated by measuring changes in both endogenous GAs and in the metabolism of exogenous [³H]- and [²H₂]GA₂₀ in the hypocotyis of sunflower (Helianthus annuus L. cv Delgren 131) seedlings exposed to ethylene. The major biologically active GAs identified by gas chromatography-mass spectrometry were GA₁, GA₁₉, GA₂₀, and GA₄₄. In hypocotyls of seedlings exposed to ethylene, the concentration of GA₁, known to be directly active in regulating shoot elongation in a number of species, was reduced. Ethylene treatment reduced the metabolism of [³H]GA₂₀ and less [²H₂]GA₁ was found in the hypocotyls of those seedlings exposed to the higher ethylene concentrations. However, it is not known if the effect of ethylene on GA₂₀ metabolism was direct or indirect. In seedlings treated with exogenous GA₁ or GA₃, the hypocotyls elongated faster than those of controls, but the GA treatment only partially overcame the inhibitory effect of ethylene on elongation. We conclude that GA content is a factor which may limit elongation in hypocotyls of sunflower, and that while exposure to ethylene results in reduced concentration of GA₁ this is not sufficient per se to account for the inhibition of elongation caused by ethylene.     

Endogenous Gibberellins in Elongating Shoots of Clones of Salix dasyclados and Salix viminalis

Elongating shoots of rapidly growing clones of Salix viminalis L. (clone 683-4) and Salix dasyclados Wimm. (clone 908) harvested in early August were analyzed for endogenous gibberellins (GA). Distribution of GA-like activity, determined by Tan-ginbozu dwarf rice microdrop bioassay after reverse phase C₁₈ high performance chromatography, was similar for both species. For S. dasyclados, combined gas chromatography-selected ion monotoring (GC-SIM) yielded identifications of GA₁, GA₈, GA₁₉, GA₂₀, and GA₂₉. Identifications of GA₄ and GA₉ were also made using co-injections of known amounts of [17, 17-²H₂]GAs. By bioassay, the main activity was GA₁₉-like in both species. Gibberellin A₁, GA₁₉, and GA₂₀ concentrations were approximated by GC-SIM using co-injections of known amounts of [17,17-²H₂]GAs. Both bioassay and GC-SIM results indicated very high concentrations of GA₁₉ and GA₂₀ (about 6000 nanograms per kilogram fresh weight shoot tissue using GC-SIM: 800 ng using bioassay), compared to the concentration of GA₁ (about 130 nanograms per kilogram fresh weight using either GC-SIM or bioassay).     

Gibberellins and leaf expansion in near-isogenic wheat lines containing Rht1 and Rht3 dwarfing alleles

In near-isogenic lines of winter wheat (Triticum aestivum L. cv. Maris Huntsman) grown at 20° C under long days the reduced-height genes, Rht1 (semi-dwarf) and Rht3 (dwarf) reduced the rate of extension of leaf 2 by 12% and 52%, respectively, compared with corresponding rht (tall) lines. Lowering the growing temperature from 20° to 10° C reduced the rate of linear extension of leaf 2 by 2.5-fold (60% reduction) in the rht3 line but by only 1.6-fold (36% reduction) in the Rht3 line. For both genotypes, the duration of leaf expansion was greater at the lower temperature so that final leaf length was reduced by only 35% in the rht3 line and was similar in the Rht3 line at both temperatures. Seedlings of the rht3 (tall) line growing at 20° C responded positively to root-applied gibberellin A₁ (GA₁) in the range 1–10 μM GA₁; there was a linear increase in sheath length of leaf 1 whereas the Rht3 (dwarf) line remained unresponsive. Gibberellins A_{1, 3, 4, 8, 19, 20, 29, 34, 44} and ₅₃ were identified by full-scan gas chromatography-mass spectrometry in aseptically grown 4-d-old shoots of the Rht3 line. In 12-d-old seedlings grown at 20° C, there were fourfold and 24-fold increases in the concentration of GA₁ in the leaf expansion zone of Rht1 and Rht3 lines, respectively, compared with corresponding rht lines. Although GA₃ was present at a similar level to GA₁ in the rht3 (tall) line it accumulated only fivefold in the Rht3 (dwarf) line. The steady-state pool sizes of endogenous GAs were GA₁₉ ? GA₂₀ = GA₁ in the GA-responsive rht3 line whereas in the GA non-responsive Rht3 line the content of GA₁₉≈ GA₂₀ ? GA₁. It is proposed that one of the consequences of GA₁ action is suppression of GA₁₉-oxidase activity such that the conversion of GA₁₉ to GA₂₀ becomes a rate-limiting step on the pathway to GA₁ in GA-responsive lines. In the GA-non-responsive Rht lines it is suggested that GA₁₉ oxidase is not downregulated to the same extent and GA₁ accumulates before the next rate-limiting step on the pathway, its 2β-hydroxylation to GA₈. The steady-state pool sizes of GA_{19, 20, 1, 3} and ₈ were similar in developmentally equivalent tissues of the rht3 (tall) line growing at 10° C and 20° C despite a 2.5-fold difference in the rate of leaf expansion. In contrast, in the Rht3 (dwarf) line, the extent of accumulation of GA₁ reflected the severity of the phenotype at the two temperatures with slower growing tissues accumulating less, not more, GA₁. These results are interpreted as supporting the proposed model of regulation of the GA-biosynthetic pathway rather than previous suggestions that GA₁ accumulates in GA-insensitive dwarfs as a consequence of reduced growth rates.     

A Gibberellin-Deficient Brassica Mutant-rosette

A single-gene mutant (rosette [ros/ros]) in which shoot growth and development are inhibited was identified from a rapid cycling line of Brassica rapa (syn campestris). Relative to normal plants, the mutant germinated slowly, had delayed or incomplete floral development, and reduced leaf, petiole, and internode growth. The exogenous application of GA₃ by foliar spray or directly to the shoot tip of rosette resulted in rapid flowering, bolting (shoot elongation), and viable seed production. Shoots of rosette contained endogenous levels of total gibberellin (GA)-like substances (`Tan-ginbozu' dwarf rice assay) of about one-tenth of that of the normal rapid-cycling line of B. rapa which consisted almost entirely of a very nonpolar, GA-like substance which yielded GA₁ and GA₃ upon mild acid hydrolysis. In a normal rapid-cycling B. rapa line, the nonpolar putative GA₁ and GA₃ conjugates were present, but additionally, free GA₁ and GA₃ were abundant and identified by gas chromatography-mass spectrometry-selected ion monitoring. The quantities of free GA₁ and GA₃ in the normal line and in rosette were quantified by GC-MS-SIM using [²H₂]GA₁ as an internal standard. Fourteen-day-old rosette and normal seedlings contained 5.3 and 23.2 ng GA₁ per plant, respectively. At day 21 the rosette plants contained 7.7 and 26.1 nanograms per plant of GA₁ and GA₃, while normal plants contained 31.1 and 251.5 nanograms per plant, respectively. Thus, normal plants contained from four to ten times higher levels of total GA-like substances, GA₁, or GA₃, than rosette. The ros allele results in reduced GA level, yielding the rosette phenotype whose delayed germination and flowering, and reduced shoot growth responses indicate a probable role for endogenous GA₁ and GA₃ in the regulation of these processes in Brassica.     

Effect of gibberellin a(3) on the endoplasmic reticulum and on the formation of glyoxysomes in the endosperm of germinating castor bean

Seedlings of castor bean (Ricinus communis cv. Hale) were exposed to a range of concentrations of gibberellin A₃ (GA₃). Treatments for 20 hours with GA₃ concentrations of 0.5 μM or higher resulted in increased levels of NADH-cytochrome c reductase, phosphorylcholine glyceride transferase, and malate synthase in endoplasmic reticulum (ER) isolated from endosperm on linear sucrose gradients. GA₃ treatment also resulted in increased RNA associated with ER. Malate synthase and catalase in crude homogenates were enhanced by 1 to 100 μM GA₃ concentrations. Isocitrate lyase, citrate synthase, malate synthase, catalase, and glycolate oxidase in isolated glyoxysomes were enhanced by 60, 20, 18, 40, and 28%, respectively, over controls. Treatment with abscisic acid led to decreased levels of glyoxysomal enzymes and reduced glyoxysomal protein. The effect of GA₃ and abscisic acid on the specific activities of glyoxysomes of different densities suggests that GA₃ influences enzyme levels and glyoxysome assembly.     

Spore abortion index (SAI) as a promising tool of evaluation of spore fitness in ferns: an insight into sexual and apomictic species

Ferns reproduce through small and usually haploid spores. The general paradigm states that whereas species produce good shaped spores, hybrids are sterile and form aborted spores. Apomictic fern species represent an unusual case, and it is believed that they produce an unbalanced spore spectrum. Until now, no comprehensive comparison of sexual and apomictic taxa using extensive spore fitness data has been published. Based on a representative data set of 109 plants from 23 fern taxa, we accomplished the first robust analysis of spore fitness using spore abortion index (SAI), the ratio of aborted to all examined spores. One thousand spores were analyzed for each plant. Focusing mainly on two major European fern taxa (Asplenium, Dryopteris), we compared this trait for different fern reproductive types (sexual/apomicts/hybrids) and ploidy levels (diploid versus polyploid). Our results confirmed the general assumption that shows higher SAI for apomictic taxa (18%) when compared to sexual taxa (3%). Furthermore, hybrids are characterized by having almost all spores aborted (99.8%) with the notable exception of pentaploid Dryopteris × critica (93.1%), the hybrid between sexual and apomictic taxa. We found no significant difference in SAI between sexual taxa of various ploidy levels or between sexual taxa of genera Dryopteris and Asplenium. Additionally, we carried out an optimization of the SAI method, outlying important guidelines for the use of this method in the future.     

Endogenous Gibberellins of Pine Pollen: III. Conversion of 1,2-[H]GA(4) to Gibberellins A(1) and A(34) in Germinating Pollen of Pinus attenuata Lemm

Gibberellins A₁ and A₃₄ (possibly A₂) were found as products of metabolism of 1,2-[³H]GA₄ during germination of Pinus attenuata pollen. The conversion from GA₄ to GA₁ and GA₃₄ occurred as hydroxylations at atoms C-13 and C-2 of the ent-gibberellane skeleton, respectively. Percentage interconversion of the GA₄ absorbed was in the range of 0.15 to 0.43% for GA₁ and 1.54 to 3.22% for GA₃₄. Identifications were made on a gas-liquid chromatograph with radioactive monitoring by comparison with standards.     

The differential effects of C-16,17-dihydro gibberellins and related compounds on stem elongation and flowering in Lolium temulentum

Plants of Lolium temulentum L. cv. Ceres grown under short days (SDs) can be induced to initiate inflorescences either by exposure to one long day (LD) or by single applications of some gibberellins (GAs), which also enhance the flowering response to one LD. Single doses of up to 25 μg per plant of C-16, 17-dihydro-GA₅ were about as effective as GA₅ for promoting flowering after one LD but inhibited stem elongation by up to 40% over three weeks. The promotion of flowering but not the inhibition of elongation by 16, 17-dihydro-GA₅ was reduced in SDs or in LDs low in far-red (FR) radiation. With shoot apices cultured in vitro, 16, 17-dihydro-GA₅ was more florigenic than GA₃ for apices excised after one LD of 14 h or more, but less florigenic for apices excised from plants in shorter days. 16, 17-Dihydro-GA₅ was ineffective compared with GA₁, GA₃ and GA₅ for α-amylase production by half-seeds of Lolium, a response concordant with its effect on stem elongation. As with GA₅, 16, 17-dihydro derivatives of GA₁, GA₃, GA₂₀ and several other GAs were more effective for flowering and less effective for stem elongation than the GAs from which they were derived. Hydroxylation at C-17 and/or C-16 generally reduced the effectiveness of 16, 17-dihydro-GA₅ for flowering. These results extend the known features of GA structure which favour flowering relative to stem elongation in L. temulentum. Moreover, C-16, 17-dihydro-GA₅ mimics, in its daylength- and wavelength-dependence and lack of stem elongation, characteristics of the LD stimulus in L. temulentum.     

Shoot elongation in Lathyrus odoratus L.: Gibberellin levels in light- and dark-grown tall and dwarf seedlings

The levels of gibberellin A₁ (GA₁), GA₂₀, GA₁₉, GA₈, GA₂₉ and GA₈₁ (2-epiGA₂₉) were measured in tall (L-) and dwarf (ll) sweet-pea plants grown in darkness and in light. In both environments the apical portions of dwarf plants contained less GA₁; GA₈ and GA₁₉, but more GA₂₀, GA₂₉, and GA₈₁ than did those of tall plants. It is concluded that the partial block in 3β-hydroxylation of GA₂₀ to GA₁ is imposed by allele l in darkness as well as in the light. Furthermore, darkness does not appear to enhance elongation in sweet pea by increasing GA₁ levels. The reduction of the pool size of GA₁₉ in dwarf plants supports recent theories on the regulation of GA biosynthesis, formulated on the basis of observations in monocotyledonous species. Darkness results in decreased GA₂₀, GA₂₉, and GA₈₁ levels in the apical portions of tall and dwarf plants and possible reasons for this are discussed.