Analysis of Phytohormone Profiles during Male and Female Cone Initiation and Early Differentiation in Long-shoot Buds of Lodgepole Pine

In lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm), cone initiation and gender differentiation are site-specific in long-shoot buds, with female cones in the distal portion and male cones in the proximal portion. By using high-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC–ESI–MS/MS) in multiple-reaction monitoring (MRM) mode, cytokinins, indole-3-acetic acid (IAA), abscisic acid (ABA), and their selected metabolites were investigated in developing long-shoot buds from multiple genotypes. Spatially, higher concentrations of trans-zeatin riboside (t-ZR) and dihydrozeatin riboside (dhZR) existed in the distal parts of long-shoot buds, whereas concentrations of isopentenyl adenosine (iPA), IAA, ABA glucose ester (ABA-GE), and phaseic acid (PA) were higher in the proximal parts in all investigated genotypes. In long-shoot buds of genotypes with a history of high female cone yield, concentrations of t-ZR and the ratio of zeatin-type to isopentenyl-type cytokinins were higher in the entire buds, whereas dhZR or IAA was higher in either the distal or the proximal part, respectively. In low female cone yielding genotypes, concentrations of c-ZR, iPA, ABA-GE, and PA were higher in both of the parts. Temporally, concentrations of several hormone-related compounds showed obvious changes in late June and late July, prior to male and female cone bud differentiation. This study reveals that the local hormonal status in a long-shoot bud at specific developmental stages may play an important role in gender determination and cone yield.     

Comparison of endogenous cytokinins, ABA and metabolites during female cone bud differentiation in low and high cone-producing genotypes of lodgepole pine

In lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.), cone bud initiation within long-shoot buds varies according to genotype. We chose to study hormone profiles of two genotypes that differed significantly in cone yield. Phytohormone profiles were established by high performance liquid chromatography–electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode with samples collected from genotypes 299 and 233, the typically high and low cone producers. Generally, concentrations of trans-zeatin-O-glucoside were higher in genotype 299, whereas dihydrozeatin concentrations were higher in genotype 233. Both isopentenyl adenine and isopentenyl adenosine were present at higher concentrations in genotype 233. The ratio of total quantifiable zeatin (Z)-type cytokinins to isopentenyl (iP)-type cytokinins was approximately threefold higher in genotype 299 during female cone bud differentiation. In genotype 299, ABA concentration was significantly lower than in genotype 233 on the first sampling date, while the phaseic acid concentration was lower consistently throughout the period investigated. Dihydrophaseic acid was present in low concentrations in most samples of genotype 233, but was not quantifiable in genotype 299. Our study reveals that long-shoot buds of the high cone-producing genotype had higher ratios of Z-type cytokinins to iP-type cytokinins than were found in the low cone-producing genotype. High cone-producing buds also contained less ABA, phaseic acid and dihydrophaseic acid during female cone bud differentiation.     

Cytokinins in Vegetative and Reproductive Buds of Pseudotsuga menziesii

Immunoaffinity techniques using columns of immobilized antibodies raised against zeatin riboside and isopentenyladenosine were found to be effective in isolating cytoklnins from vegetative, female, and male buds of Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco). The purified cytokinins were separated by reverse phase high performance liquid chromatography and analyzed by radioimmunoassay. Confirmation of cytokinin identities was by gas chromatography-mass spectrometry. Immediately prior to bud burst, all bud types contained three major cytokinins: isopentenyladenosine, zeatin riboside, and a hexose conjugate of zeatin riboside (not zeatin riboside O-glucoside). Zeatin-type cytokinins were present in relatively high concentration in vegetative and female buds. In male buds, however, relatively high levels of isopentenyladenosine were found together with low levels of zeatin-type cytokinins.     

Promotion of flowering in the Pinaceae by gibberellins

Significant male and female flowering (cone bud production) by girdled branches of 6-year-old Douglas fir (Pseudotsuga menziesii (Mirb. Franco) seedlings was promoted by applications (mid-April to June) of 1.6 or 3.2 mg per branch (in total) of certain non-polar gibberellins (GA's). Girdling alone was ineffective. When tested alone, a mixture of GA_4/7 was most effective. GA₉ less so, while GA₅ and the more polar GA₃ were essentially ineffective. For female cone buds GA_4/7+ GA₉ were synergistically effective, but for male cone buds GA_4/7 alone was best. The auxin naphthaleneacetic acid (NAA) was not tested alone, but at low dosage (0.175 mg/branch in total) NAA enhanced the flowering efficacy of GA's for both sexes; at a high dosage (0.875 mg/branch in total) male cone bud production was further enhanced, but only at the expense of females. For female flowering the best treatment (90% frequency of flowering 6.8 cone buds/branch), was GA_4/7+ GA₉+ low NAA; for male flowering, it was GA_4/7+ high NAA (30% frequency and 4.2 cone buds/branch. Frequency of flowering for controls was 18% and 0%, average number of cone buds/branch was 0.9 and 0, for females and males, respectively. The successful treatments did not affect promordia initiation, rather they caused the differentiation of previously initiated, but undetermined, lateral primordia into cone and latent buds at the expense of vegetative bud differentiation. The lack of success reported by earlier workers in promoting flowering in Pinaceae species by GA's appears to be the unfortunate result of selecting GA₃ for initial testing. The practical implications of this early and enhanced flowering by non-polar GA's seedlings of a commercially important conifer are discussed in relation to accelerating the processes of tree improvement.     

Gibberellin A(4/7) and the Promotion of Flowering in Pinus radiata: Effects on Partitioning of Photoassimilate within the Bud during Primordia Diferentiation

Gibberellin A_4/7 mixture (GA_4/7), a highly effective promoter of early and enhanced flowering in the Pinaceae, caused a significant reallocation of dry matter and ¹⁴C-photosynthate within terminal buds of Pinus radiata D. Don within 8 days of hormone treatment. Treatment with GA_4/7 to terminal shoots of vigorous, potentially flowering mature grafted propagules reduced the flow of photoassimilated ¹⁴C and dry matter into the terminal bud as a whole, but significantly increased the dry matter and ¹⁴C allocated within the bud to developing long-shoot primordia (potential seed-cone buds). This was accomplished at the expense of the structural tissues, the apical dome region, and the vegetative branch buds. Although GA₃ caused a similar reallocation of dry matter within the terminal bud, it was significantly less effective than GA_4/7 thus appears to have, in addition to any nutrient diversion abilities, a distinct morphogenic function in sexual differentiation.     

Hormonal Profile in Vegetative and Floral Buds of Azalea: Levels of Polyamines, Gibberellins, and Cytokinins

The floral transition includes a complex system of factors that interact and involve various biochemical signals, including plant growth regulators. The physiological signals involved in the control of the floral transition have been sparsely studied and mainly in plant species whose genetics are poorly known. In this work, the role of polyamines, gibberellins, and cytokinins was investigated by analyzing their endogenous content in vegetative and floral buds of azalea. The results showed that there is a clear distinction between floral and vegetative buds with respect to the levels of these plant hormones, with floral buds containing higher amounts of conjugated polyamines, gibberellins (GAs) from the non-13-hydroxylation pathway (GA₉, GA₇, and GA₄), and cytokinins (particularly isopentenyl-type species), and vegetative buds containing higher amounts of free polyamines and gibberellins from the early 13-hydroxylation pathway and fewer cytokinins. In conclusion, there is a specific pattern of endogenous hormone profiles in both vegetative and floral bud development in azalea, which may be relevant for future research on the control of flowering by exogenous hormone applications.     

In vitro induction of tetraploid <Emphasis Type="Italic">Ziziphus jujuba</Emphasis> Mill. var. <Emphasis Type="Italic">spinosa</Emphasis> plants from leaf explants

The genetic manipulation of Capsicum has been unsuccessful, and a large bottleneck to transferring the desired genes is due to the difficulty in regenerating whole plants through tissue culture because of its highly recalcitrant and high genotype specificity. This study aimed to investigate and establish rapid shoot regeneration from the proximal ends of the leaves of Capsicum frutescens KT-OC and BOX-RUB varieties. A maximum of 8–10 shoot buds were obtained from the margins of the proximal portion of a cotyledonary leaf explant of C. frutescens variety KT-OC on medium I containing 44.44 µM 6-benzylaminopurine (BA), 5.71 µM indole-3-acetic acid (IAA), 10 µM silver nitrate (AgNO₃) and 1.98 mg L^?1 2-(N-morpholine) ethane sulphonic acid within 4 weeks of incubation, of which 60% of explants responded in terms of shoot buds. Petiole explants (40%) cultured on the same medium produced 2–4 shoots per explant from the distal portion. The cut portions of the cotyledonary leaf proximal portions responded well to shoot bud formation in the presence of 22.20 µM BA and 14.68 µM phenyl acetic acid (PAA), wherein 100% of explants responded in terms of shoot bud formation, with an average of 10?±?1.7 and 8?±?1.9 shoot buds per explant in KT-OC and BOX-RUB varieties, respectively. The differentiated shoots grew well and proliferated in the presence of 14.68 µM PAA?+?22.20 µM BA and 10 µM AgNO₃. Shoot elongation was obtained in presence of 1.44 µM gibberellic acid (GA₃) and 10 µM AgNO₃. These shoots were rooted on plant growth regulator-free half-strength MS medium and upon hardening; field survival rate was 70%. This reproducible regeneration method for C. frutescens, especially the Indian high pungent variety, from proximal portion of cotyledonary leaf and petiole explants, can be used for biotechnological improvement.     

Changes in Cytokinins before and during Early Flower Bud Differentiation in Lychee (Litchi chinensis Sonn.)

Lychee (Litchi chinensis) has been analyzed for cytokinins in buds before and after flower bud differentiation, using reversephase high performance liquid chromatography in combination with Amaranthus bioassay and gas chromatography-mass spectrometry-selected ion monitoring. Four cytokinins, zeatin, zeatin riboside, N⁶-(δ²-isopentenyl)adenine, and N⁶-(δ⁶-isopentenyl) adenine riboside, were detected in buds. There was an increase of cytokinin activity in the buds during flower bud differentiation. In dormant buds, the endogenous cytokinin content was low, and the buds did not respond to exogenous cytokinin application. Application of kinetin promotes flower bud differentiation significantly after bud dormancy. These results are interpreted as an indication that the increase in endogenous cytokinin levels during flower bud differentiation may be correlative rather than the cause of flower bud initiation.     

Protoplast fusion in the genus <Emphasis Type="Italic">Gentiana</Emphasis>: genomic composition and genetic stability of somatic hybrids between <Emphasis Type="Italic">Gentiana kurroo</Emphasis> Royle and <Emphasis Type="Italic">G. cruciata</Emphasis> L.

Thidiazuron [N-phenyl-N-(1, 2, 3-thidiazol-5-yl) urea, TDZ] treatment significantly improved shoot morphogenesis of Saussurea involucrata Kar. Et Kir (S. involucrata) leaf explants. The biochemical mechanisms underlying TDZ-induced shoot organogenesis were investigated by measuring endogenous plant growth hormones, H₂O₂, as well as the activities of superoxide dismutase (SOD) and catalase (CAT). The levels of endogenous gibberellic A3 (GA₃) and zeatin (ZT) significantly increased in leaf explants subject to a 28-day treatment than the controls. However, extending exposure time to TDZ inhibited GA₃ accumulation. At the same time, the SOD activity increased significantly until the 28th of TDZ treatment time and the CAT activity reduced simultaneously, which was closely linked with the significant increase in H₂O₂ concentrations in the explants. And there was a sharply promotion after the 35 day of culture time if the plant tissue was always in medium contained, which was in company with the cell activity decreased. We propose that a combination of increased GA₃, ZT, and H₂O₂ concentration is the basis for the enhanced shoot morphogenesis in response to TDZ treatment. These results provide a starting point for an improved understanding of the biochemical mechanisms underlying TDZ-induced shoot organogenesis of S. involucrata.     

Interaction of Phytohormones in Regulating the Axillary Bud Growth in Pea

In the present study the interactions of GR24, a synthetic analog of newly discovered plant hormones strigolactones (SLs), with cytokinin (CK), benzyladenine (BA), auxin naphthaleneacetic acid (NAA), gibberellic acid (GA₃) and abscisic acid (ABA) in the regulation of axillary bud growth in pea (Pisum sativum L.) were investigated. The hormones were applied directly to buds at node 1 and 2 and the dose-response experiments were performed on 8–10-day-old SL-deficient rms1 and rms5–1 mutants, branching SL-sensitive rms2–1 mutants and wild-type plants. In the mutant plants the treatment with 5 μM GR24 completely inhibited bud growth while BA up to 100 μM stimulated it. The combined application of GR24 and BA showed that GR24 efficiency to reduce bud outgrowth constantly declines as CK-stimulated bud growth increased, with the inhibiting effect of GR24 abolished at 100 μM BA applied. GA₃ accelerated bud outgrowth, but did not interfere with GR24 inhibitory action. NAA reduced bud growth in intact SL-sensitive rms2–1 mutant and also in SL-insensitive rms3–2 and rms4–1 mutants. The NAA effect was observed already at 0.5 μM, however, even at a response saturating concentration of 500 μM its inhibiting effect was inferior to that of 5 μM GR24. At combined treatment the effectiveness of GR24 in suppressing bud growth decreased with a decrease in NAA-inhibited bud growth, suggesting that auxin might act as a suppressor of SL action. ABA strongly inhibited the bud outgrowth and, like CK and auxin, reduced the inhibitory effectiveness of GR24. The revealed ability of CK, ABA and auxin to suppress bud response to SLs is supposed to result from phytohormone signaling crosstalks.     

Adenine type and diphenyl urea derived cytokinins improve the postharvest performance of <Emphasis Type="Italic">Iris germanica</Emphasis> L. cut scapes

An experiment was designed to evaluate the effect of various adenine derived cytokinins (kinetin and 6-benzylaminopurine) and diphenyl urea cytokinin (thidiazuron) on the postharvest performance of cut scapes of Iris germanica. Flower scapes were harvested with the oldest bud at ‘1 day before anthesis stage’, brought to laboratory under water, cut to a uniform length of 35 cm, divided into three sets viz., kinetin (KIN), 6-benzyl aminopurine (BAP) and thidiazuron (TDZ). Each set of scapes was treated with a particular cytokinin alone or in combination with 0.1 M sucrose. TDZ was effective than KIN and BAP in improving the postharvest life of the I. germanica scapes by 5.4 days as compared to the control (untreated scapes held in distilled water). This was because of the minimum percentage of bud abortion by TDZ application. Cytokinin application resulted in increased antioxidant activity, higher protein and phenolic content, besides a decrease in specific protease activity and α-amino acids in the tepal tissues. Application of TDZ resulted in the maximum increase in the superoxide dismutase, catalase and ascorbate peroxidase activity in the tepal tissues. The scapes treated with BAP and KIN maintained higher carbohydrate content in the tissue samples as compared to control and TDZ treated scapes. TDZ and BAP application resulted in increased membrane stability because of the decreased lipoxygenase activity which prevented membrane lipid peroxidation. Among the cytokinins tested, TDZ proved to be the promising cytokinin in improving the postharvest performance of beautiful flowers of I. germanica scapes.     

Fluctuation in levels of endogenous plant hormones in ovules of normal and mutant cotton during flowering and their relation to fiber development

The contents of indole-3-acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA), and cytokinins were determined in ovules of normal cotton (Tm-1) and a kind of fiber differentiation mutant (Xin) before and after flowering by enzyme-linked immunosorbent assays. It was found that 24 h before flowering, a peak of IAA content was observed in ovules of Tm-1, whereas in ovules of Xin, a low level of IAA was determined. From –1 day (1 day before flowering) to +3 days (3 days after flowering), GA₁₊₃ levels in ovules of Xin were 40–70% lower than those of Tm-1; GA₄₊₇ levels were very low, and there was no visible difference in GA₄₊₇ content between normal and mutant cotton. The ABA content in ovule of Tm-1 decreased by 70% 3 days after flowering, whereas that of Xin only decreased by 20%. The levels of cytokinins in ovules of Tm-1 decreased after flowering, and those of Xin kept up a steady increase.Abbreviations IAA indole-3-acetic acid - GA gibberellin - ABA abscisic acid - ELISA enzyme-linked immunosorbent assay - FW fresh weight - PBS phosphate-buffered saline - iPA isopentenyladenosine - ZR zeatin riboside - DHZR dihydrozeatin riboside - CTK cytokinin     

Somatic embryogenesis of <Emphasis Type="Italic">Byrsonima intermedia</Emphasis> A. Juss.: induction and maturation via indirect approach

Byrsonima, especially the species Byrsonima intermedia, is an endangered Brazilian plant that has been widely used as food and for its therapeutic characteristics. However, this species faces challenges with sexual propagation, and somatic embryogenesis has emerged as a viable alternative option for propagation. Therefore, this study aimed to establish a protocol for inducing somatic embryogenesis in B. intermedia. For the induction of callus from in vitro seedling leaves, different subcultures (three subcultures, 60 days each) and concentrations of different cytokinins (BAP, TDZ, Kin and ZEA) combined with varying NAA solutions were tested. Different concentrations of NAA were also analyzed in the induction of pro-embryogenic calli. For the induction of embryogenic calli and somatic embryos, the pro-embryogenic calli were subcultured on MS medium without adding growth regulators. The somatic embryos that originated were inoculated on a maturation medium containing different concentrations of gibberellic acid (GA₃). The formation of secondary embryos was also analyzed using different concentrations (0, 2.88, and 8.66 µM) of GA₃ and different types of lids (Conventional lid, Biossama® commercial lid and conventional lid with membranes). The results show that for the induction of somatic embryos, the use of kinetin with NAA presented the formation of somatic embryos in the second (4.76 µM CIN?+?0.54 µM NAA) and third (5.17 µM CIN?+?10.54 µM NAA) subcultures. The use of 28.87 µM GA₃ favored the formation of seedlings. The Biossama lid and 2.88 µM GA₃ showed higher formation of secondary embryos.     

Effect of gibberellin on <Emphasis Type="Italic">Arabidopsis</Emphasis> growth,development, and hormonal balance under green and blue light

The role of gibberellins in plant morphology under selective light was studied. A comparison of the effects of green and blue light on growth, development, and hormonal balance was performed for two Arabidopsis thaliana (L.) Heynh. (ecotype Landsberg erecta lines: wild type Ler and its ga4-1 mutant with suppressed GA_4/1 synthesis. The absence of active GA_4/1 from ga4-1 mutant determined its retarded growth, slowed passing through developmental phases, suppressed apical dominance, and reduced seed productivity. The retarded growth and development of the mutant was related to changed hormonal balance in them as compared to wild-type line: IAA content and the IAA/ABA ratio were declined, zeatin riboside and ABA accumulated. Green light retarded stem elongation and branching, reduced leaf specific surface density and plant seed productivity, and retarded plant transition to reproduction to a greater degree at GA_4/1 deficit (ga4-1) than at its normal content (Ler).     

Factors influencing direct shoot regeneration from mature leaves of Jatropha curcas, an important biofuel plant

In order to establish a highly efficient and sustainable regeneration system, we systematically researched the key factors affecting direct shoot regeneration from Jatropha curcas leaves that were collected from Hainan (HN1-1), Lijiang (LJ3-1), and Yuxi (YX2-12) provinces in China. The L₉(3⁴) orthogonal test of thidiazuron (TDZ), kinetin (Kn), and gibberellic acid (GA₃) were studied, and the explant type, growth age, and cultivar of leaves were subsequently investigated. Simultaneously, the combinations of plant growth regulators (PGRs) promoting shoot bud proliferation, elongation, and root establishment were examined. The results showed that the best medium for shoot bud induction was Murashige and Skoog (MS) medium supplemented with 1.0 mg/L TDZ, 0.5 mg/L Kn, and 0.5 mg/L GA₃. TDZ was the key PGR, while Kn and GA₃ played an important role in shoot bud elongation and the number of shoots per leaf disk, respectively. The induced shoot buds proliferated and readily elongated in MS medium with 0.3 mg/L 6-benzylaminopurine and 0.01 mg/L indole-3-butyric acid (IBA) and established roots in half-strength MS medium supplemented with 2.0 mg/L IBA. Using the previously described methods, the third to fifth leaves were found to be the best explant source for shoot bud induction, with a high induction rate, large shoot numbers per disk, excellent proliferation, and consistent rooting. With the use of this regeneration system, the shoot bud induction rate increased from the reported rate of 53.5% to more than 90% using different explants and cultivars, and the shoot number per leaf disk (shoot length?≥?0.5 cm) increased from 1.6 to 3.5. Thus, this optimized regeneration system will effectively promote the propagation and genetic transformation of J. curcas.     

The content of endogenous hormones and sugars in the process of early somatic embryogenesis in the tree fern <Emphasis Type="Italic">Cyathea delgadii</Emphasis> Sternb.

Somatic embryogenesis (SE) of Cyathea delgadii presents a model system for investigating the mechanisms associated with the acquisition of embryogenic competence by single epidermal cells of stipe explants cultured on plant growth regulator-free medium. The present work reveals relationship between endogenous hormone and sugar content in the process of early SE in C. delgadii. By comparing two types of initial explants, i.e. incapable (non-etiolated) and capable (etiolated) of SE, it was established that in etiolated explants, the glucose, fructose, sucrose, and abscisic acid (ABA) contents diminished, but indole-3-acetic acid (IAA) and cytokinins (CKs; i.e. cis/trans zeatin, cis/trans-zeatin riboside, kinetin, kinetin riboside, isopentenyladenosine) contents increased. The ratios between phytohormones revealed that a high concentration of ABA is the main factor inhibiting SE induction. Because of explant excision, a dramatic reduction in concentration of all phytohormones studied was observed, but hormonal balance and sugar content remained almost unchanged. During the 14-day-long culture, the ABA/CKs and ABA/IAA ratios remained constant, whereas the greatest differences were detected for the IAA/CKs and Z-type/iPA cytokinin ratios. Excluding day 6 of culture, cytokinins were found to be the predominant phytohormones over IAA. An almost 12-fold increase in soluble sucrose concentration at day 6 of culture might be the switch to the SE expression phase. Frequent cell divisions leading to somatic embryo formation are clearly associated with increase in trans-zeatin riboside content.     

Gibberellin-induced delay of leaf senescence of Alstroemeria cut flowering stems is not caused by an increase in the endogenous cytokinin content

The effects of various chemically pure gibberellins and cytokinins on leaf yellowing of Alstroemeria were described. The loss of chlorophyll was measured both in leaves of cut flowering stems and in a model system consisting of detached leaf tips. It was demonstrated that plant growth substances affected chlorophyll loss in both systems to the same extent. Leaf senescence was delayed by various gibberellins and cytokinins. The results demonstrated that some of the gibberellins (GA₄ and GA₇) are far more effective in delaying chlorophyll loss than GA₃, which is commonly used as a postharvest treatment for Alstroemeria cut flowering stems. Immunoassays were used to demonstrate that the effect of gibberellins on leaf yellowing does not involve an increase in the endogenous cytokinin concentrations in the leaves as an intermediate step.Abbreviations GA gibberellin A - HPLC high performance liquid chromatography - GA_3Mc GA₃-methyl ester - ZR zeatin riboside - IPAR isopentenyl adenine riboside.     

枣花芽分化过程中营养物质和内源激素含量及抗氧化酶活性变化研究

以新疆主栽品种灰枣和骏枣的花芽为材料,测定不同分化时期花芽的可溶性糖、还原糖、淀粉、可溶性蛋白含量,SOD、POD、PPO、CAT活性以及内源GA3、IAA、ABA、ZT水平的变化,并分析它们与花芽分化的关系,为枣花芽分化调控提供理论参考.结果表明:(1)灰枣和骏枣花芽可溶性糖、还原糖和淀粉含量在花芽分化过程的变化趋势...     

Effect of gibberellic acid on the cyanobacterium <Emphasis Type="Italic">Nostoc linckia</Emphasis>

We investigated the influence of gibberellic acid (GA₃; 0, 1, 10, and 100 μM) on Nostoc linckia culture at 7, 14, and 21 days. The fresh and dry weight of N. linckia was increased considerably by the 10 and 100 μM GA₃ treatments. A reduction in heterocyst frequency was observed in cultures treated with 1 and 10 μM GA₃. Adding GA₃ to N. linckia culture had a little effect on cell size. The amount of chlorophyll a and carotenoids decreased at all concentrations of GA₃. The amount of phycocyanin increased up to twofold in 7-day-old culture treated with 1 μM GA₃, and similar changes were observed for allophycocyanin and phycoerythrin content after 7 days. The effect of GA₃ on reducing sugar content was different and was dependent on the growth period. A reduction in soluble sugar content was detected after GA₃ application in 7- and 14-day-old cyanobacteria. Cultures treated with GA₃ had a higher protein content after 14 days and a lower protein content after 7 and 21 days, and reduced nitrogenase activity after 7, 14, and 21 days. Our data show that GA₃ application can be a suitable and inexpensive way to increase N. linckia biomass and phycobiliprotein production.     

The effect of exogenous and endogenous phytohormones on the in vitro development of gametophyte and sporophyte in <Emphasis Type="Italic">Asplenium nidus</Emphasis> L.

The fern Asplenium nidus L. is in great demand as an ornamental plant. The aim of this work was to investigate the influence of phytohormones in promoting a gametophytic and sporophytic growth in homogenized sporophytes tissue. Exogenous application of 0.5 and 5 μM N ⁶-benzyladenine, 0.05 and 0.5 μM indole-3-acetic acid (IAA), and 0.3 and 3 μM gibberellic acid (GA₃) favoured sporophyte regeneration, whereas gametophyte regeneration took place when plant material was cultured in a hormone-free liquid MS medium. The endogenous contents of the auxin IAA, the cytokinins trans-zeatin, trans-zeatin riboside, dihydrozeatin, dihydrozeatin riboside, isopentenyladenine and isopentenyladenosine, and the gibberellins GA₁, GA₃, GA₄, GA₇, GA₉ and GA₂₀ in growing gametophytes and sporophytes were evaluated. Similar levels of the auxin and cytokinins and qualitative differences in the gibberellins were found between both generations.