Effect of different benzyladenine time pulses on the endogenous levels of cytokinins,indole-3-acetic acid and abscisic acid in micropropagated explants of Actinidia deliciosa

Actinidia deliciosa apical shoots were cultured in MS liquid medium with different benzyladenine (BA) pulses and using cellulose plugs as support for the explants. Abscisic acid (ABA), indole-3-acetic acid (IAA) and some cytokinins (Cks) [zeatin (Z), dihydrozeatin (DHZ), zeatin riboside (ZR), dihydrozeatin riboside (DHZR), N⁶-isopentenyladenine (iP) and N⁶-isopentenyladenosine (iPR)] levels were measured in leaves from explants cultured both in the absence and in the presence of BA (4.4 μM) for 30 min, 1, 2 and 35 d. Analyses were carried out after three subcultures at the end of the multiplication phase and after 31 d in ex vitro conditions. A clear relationship between the endogenous content of phytohormones and the growth characteristic of kiwifruit microplants could be confirmed. Microplants with the best growth characteristic were those cultured in presence of BA during 1 d. They were characterised by higher contents of IAA than the others studied, as well as by higher values in the IAA/Cks and IAA/ABA ratios, both at the end of the multiplication phase and after the acclimatisation period. Taken all together, these ratios at the end of the multiplication phase could be used as growth indicators of A. deliciosa explants behaviour under ex vitro conditions.     

Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress

Osmotic stress and endogenous hormone levels may have a role in shoot organogenesis, but a systematic study has not yet to investigate the links. We evaluated the changes of the endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) levels in rice (Oryza sativa L. cv. Tainan 5) callus during shoot organogenesis induced by exogenous plant growth regulator treatments or under osmotic stress. Non-regenerable callus showed low levels of endogenous ABA and IAA, with no fluctuation in level during the period evaluated. The addition of 100 μM ABA or 2 mM anthranilic acid (IAA precursor) into Murashige and Skoog basal induction medium containing 10 μM 2,4-D enhanced the regeneration frequency slightly, to 5 and 35%, respectively, and their total cellular ABA or IAA levels were increased significantly, correspondingly to the treatments. However, the regeneration frequency was greatly increased to 80% after treatment with 0.6 M sorbitol or 100 μM ABA and 2 mM anthranilic acid combined. Both treatments produced high levels of total cellular ABA and IAA at the callus stage, which was quickly decreased on the first day after transfer to regeneration medium. Thus, osmotic stress-induced simultaneous accumulation of endogenous ABA and IAA is involved in shoot regeneration in rice callus.     

Endogenous hormonal content and somatic embryogenic capacity of Corylus avellana L. cotyledons

Endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinins [zeatin (Z) zeatin riboside, dihydrozeatin, dihydrozeatin riboside, N⁶-isopentenyl adenine (iP) and N⁶-isopentenyladenine riboside] were evaluated in hazelnut (Corylus avellana L.) cotyledons of different developmental stage and genetic source for their somatic embryogenic capacity. There was an inverse correlation between the embryogenic potential of cotyledons and the degree of maturity of zygotic embryos, the first characteristic being associated with iP-type cytokinins and the second with Z-type cytokinins. Although the differences in total cytokinin, ABA and IAA contents between the cotyledons were small, the IAA/ABA and, mainly, the iP-type/Z-type cytokinin ratios were found to be two good indexes of the embryogenic competence of explants, suggesting that the endogenous hormonal balance is a very important factor defining the in vitro potential of hazelnut cotyledons. Received: 6 January 1997 / Revision received: 3 March 1997 / Accepted 1 April 1997     

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.     

Trichoderma harzianum strain T-22 induces changes in phytohormone levels in cherry rootstocks (Prunus cerasus?×?P. canescens)

The aim of this research was to explain the direct plant growth-promoting activity of Trichoderma harzianum strain T-22 (T22), hypothesizing the involvement of different classes of plant growth regulators. Seven days after the transfer to root-inducing medium, in vitro-cultured shoots of GiSeLa6^? (Prunus cerasus  × P. canescens) were inoculated with T22. Root and shoot growth were significantly affected by T22 (+76 and +61%, respectively). Ten days after inoculation, the levels of indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), dihydrozeatin riboside (DHZR), gibberellic acid (GA3) and abscisic acid (ABA) were analyzed by high performance liquid chromatography coupled with mass spectrometry. The results showed that after T22-inoculation, IAA and GA3 significantly increased in both leaves (+49 and +71%, respectively) and roots (+40 and +143%, respectively) whereas t-ZR decreased (−51% in leaves and −37% in roots). Changes in DHZR were observed in T22-inoculated roots (−32%) but not in leaves, whereas the levels of ABA did not differ between the two treatments. The extraction method allowed the simultaneous extraction of phytohormones. There is evidence that the change in phytohormone levels is one of the direct mechanism by which T22 promotes rooting and shoot growth, with notable advantages for rootstock production during nursery processes.     

Effect of exogenous application of IAA and GA3 on growth,protein content,and antioxidant enzymes of Solanum tuberosum L. grown in vitro under salt stress

Indole-3-acetic acid (IAA) and gibberellic acid (GA₃) are essential for the growth and development of plants. In the present study, the ameliorative potential of these phytohormones on growth, protein content, and antioxidant enzymes was investigated in in vitro-grown Solanum tuberosum L. cultivars ‘Cardinal’ and ‘Desiree’ under salt stress. A 4 × 3 factorial combination of 0, 40, 60, or 80 mM NaCl with 0, 7, or 14 μM IAA, or 0, 14, or 21 μM GA₃, were added to Murashige and Skoog (MS) basal medium, followed by inoculation of nodal explants or callus cultures. The data for root and shoot number and length, number of nodes and leaves, fresh weight of plants, increase or decrease in fresh weight of callus cultures, total soluble protein, and superoxide dismutase (SOD) and peroxidase (POD) activities were recorded after 30 d. The growth of both callus cultures and nodal explants subjected to NaCl stress was substantially reduced compared with the control. Both IAA and GA₃ successfully alleviated the harmful effects of salt stress on all of the growth parameters studied. Salt stress resulted in decreased protein content, which increased when the media also contained phytohormones. The activities of SOD and POD were increased with either IAA or GA₃ under NaCl stress. Therefore, the exogenous application of both IAA and GA₃ not only played a positive role in terms of in vitro potato growth but also significantly affected the biochemical parameters tested.

     

Micropropagation of Salix pseudolasiogyne from nodal explants

The effect of phytohormones on the breaking of dormancy of axillary buds in Salix pseudolasiogyne and their subsequent proliferation from nodal explants were examined. Nodal explants obtained from a 20–year-old S. pseudolasiogyne tree were cultured either on woody plant basal medium (WPM) or WPM supplemented with benzyladenine (BA, 2.2/4.4 μM), zeatin (1.1/2.2 μM), gibberillic acid (GA₃, 2.9 and 14.5 μM), and GA₃ + BA (2.9 + 4.4 μM). Although axillary shoots developed in all the media, a higher percentage bud break occurred on BA supplemented media. To corroborate the results, endogenous levels of cytokinins [Cks, N ⁶-isopentenyladenine (iP), zeatin riboside (t-ZR), dihydrozeatinriboside (DHZR)] and abscisic acid (ABA) were determined. On BA supplemented media, the levels of zeatin type (Z-type) of Cks were higher than those of isopentenyladenine type of Ck in the explants, while the ABA level was low. Axillary shoots did not grow well and became necrotic upon subculture to fresh basal WPM. In order to improve shoot growth, they were subcultured twice at a 4-week interval on to WPM supplemented with BA (2.2/4.4 μM), GA₃ (1.4 μM), or GA₃ + BA (1.4 + 4.4/2.9 + 4.4 μM). Maximal shoot growth (93%) was achieved on WPM supplemented with 2.2 μM BA. Comparative analyses of endogenous Cks revealed that higher Cks (Z-type Cks) were present in actively growing shoots. Rooting was readily achieved when the shoots were subcultured to WPM without phytohormones. The rooted plants were acclimatized well upon transplantation.     

Sex‐specific floral morphology,biomass, and phytohormones associated with altitude in dioecious Populus cathayana populations

Relationships between sex‐specific floral traits and endogenous phytohormones associated with altitude are unknown particularly in dioecious trees. We thus examined the relationships between floral morphology or biomass and phytohormones in male and female flowers of dioecious Populus cathayana populations along an altitudinal gradient (1,500, 1,600, and 1,700 m above sea level) in the Xiaowutai Nature Reserve in northern China. The female and male flowers had the most stigma and pollen at 1,700 m, the largest ovaries and least pollen at 1,500 m, and the smallest ovaries and greater numbers of anthers at 1,600 m altitude. The single‐flower biomass was significantly greater in males than in females at 1,600 or 1,700 m, but the opposite was true at 1,500 m altitude. The biomass percentages were significantly higher in anthers than in stigmas at each altitude, while significantly greater gibberellin A3 (GA₃), zeatin riboside (ZR), indoleacetic acid (IAA), and abscisic acid (ABA) concentrations were found in female than in male flowers. Moreover, most flower morphological traits positively correlated with IAA in females but not in males. The biomass of a single flower was significantly positively correlated with ABA or IAA in males but negatively with ZR in females and was not correlated with GA₃ in both females and males. Our results demonstrate a distinct sexual adaptation between male and female flowers and that phytohormones are closely related to the size, shape, and biomass allocation in the pollination or fertilization organs of dioecious plants, although with variations in altitude.     

Effects of Ambiol and 2-Chlorethylphosphonic Acid on the Content of Phytohormones in Potato Leaves and Tubers

The effects of the antioxidant Ambiol and 2-chlorethylphosphonic acid (2-CEPA) on individual concentrations and concentration ratios of phytohormones, photosynthesis and photophosphorylation rates, sucrose and starch content in tubers, and plant productivity were studied in potato (Solanum tuberosum L). Ambiol increased the ratio of indoleacetic acid (IAA) to abscisic acid (ABA), IAA/ABA, and that of zeatin (Z) and zeatin riboside (ZR) to ABA, (Z + ZR)/ABA. These effects were underlain by an increase in the content of auxins and cytokinins and a decrease in ABA. Unlike Ambiol, 2-CEPA increased the level of ABA, the effect being the most pronounced in the tubers. Ambiol increased the rates of photosynthesis and noncyclic photophosphorylation in chloroplasts isolated from potato leaves. The relation of this phenomenon to auxin and cytokinin accumulation, as well as Ambiol- and 2-CEPA-induced changes in the hormonal balance of potato tubers, carbon metabolism, and plant productivity, is discussed.     

Comparison of in vitro regeneration pathways in <Emphasis Type="Italic">Punica granatum</Emphasis> L.

When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus. A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM NAA, and 6 μM giberrellic acid (GA₃). However, adding 24 μM silver nitrate (AgNO₃) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm) were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal. Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l⁻¹ sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when these were transferred to MS medium was supplemented with 60 g l⁻¹ sucrose.     

Changes in cytokinin,auxin, and abscisic acid contents in wheat seedlings treated with the growth regulator Stifun

The growth regulator Stifun at all concentrations tested (0.033, 0.33, 3.3, and 33 mg/L) affected the hormonal status of wheat seedlings (Triticum aestivum L., cv. Zhnitsa) and stimulated plant growth. This was evident in activation of cell division and elongation, as well as in the increase in shoot and root length, water content, and dry weight. Effects of Stifun on roots and shoots depended on concentration. Application of Stifun at the optimal growth-stimulating concentration (0.033 mg/L) elevated the levels of zeatin, zeatin riboside, dihydrozeatin riboside, isopentenyladenosine, and IAA in roots of 2-day-old seedlings, but reduced the ABA content. The levels of ABA, IAA, dihydrozeatin, and dihydrozeatin riboside in shoots increased, while the levels of zeatin riboside, isopentenyl adenine, and isopentenyladenosine decreased. The results indicate that the hormonal system plays a part in the plant response to growth-stimulating action of Stifun.     

Effects of Chlorocholine Chloride on Phytohormones and Photosynthetic Characteristics in Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.)

Effects of chlorocholine chloride (CCC) on phytohormones and photosynthetic characteristics of Zhongshu 3, a potato (Solanum tuberosum L.) variety widely cultivated in south China, were studied by foliar CCC application on 24 and 28 days after emergence, that is, at the tuber initiation stage. It was found that on 42 days after emergence, that is, at the tuber bulking stage, spraying CCC increased indolacetic-3-acid (IAA) and zeatin (Z) contents but decreased abscisic acid (ABA) content in leaves. The content ratios of IAA/Z, IAA/ABA, Z/ABA, and (IAA + Z)/ABA in leaves treated with CCC were higher than those of the control. CCC plays a prominent regulating role in the photosynthesis of Zhongshu 3. The net photosynthetic rate (P _n), stomatal conductance (G _s), intercellular CO₂ concentration (C _i), and transpiration rate (T _r) of treated leaves were superior to those of controls at the tuber bulking stage. CCC markedly increased tuber yield and quality. The contents of sucrose and starch in tubers treated with CCC increased at the end of the vegetation period, whereas the contents of reducing sugars and solanine decreased. CCC at 2.0 g L⁻¹ was found to be the most effective concentration. Collectively, the results of this research identify phytohomonal metabolism and photosynthetic physiology of potato leaves as processes affected early after application of CCC resulting in significantly improved increases in tuber yield and quality.     

Clonal propagation byin vitro culture of Corchorus (jute)

Callus was produced on cotyledon, shoot tip, hypocotyl and root explants of twoCorchorus species on several media. Cytokinin was necessary for callus production on cotyledon explants. BothC.olitorius genotypes produced most callus on media with zeatin and either NAA or IAA, and theC.capsularis genotype produced most callus on media with IAA and either zeatin or BA. High frequencies of regenerated shoots were obtained from shoot tip explants of both species, from the apical meristem and from callus. Media with 2.0 mg 1⁻¹ BA were superior for both species, and media with zeatin were equally good forC.capsularis only. More regeneration was obtained for all genotypes after subculture of callus on media with 2.0 mg 1⁻¹ zeatin. Cotyledon callus produced less regeneration, also with differences between genotypes; explants of both genotypes ofC.olitorius produced regeneration on a medium with NAA and zeatin, and theC.capsularis genotype produced regeneration on a medium with IAA and BA. Limited regeneration from root explant callus was obtained forC.capsularis only on medium with BA and IAA. Regeneration was not obtained from hypocotyl callus. Further regeneration of shoots of both species was obtained from secondary callus after subculture, and from nodal segments of regenerated shoots and of seedling shoots cultured on basic MS medium without growth hormones. Roots were produced on about 80% of all shoots after transference to medium with 0.2 mg 1⁻¹ IBA, and rooted plantlets survived and flowered normally after transference to compost.     

Endogenous hormone concentrations and embryogenic callus development in wheat

Immature zygotic embryos of two wheat (Triticum aestivum L.) genotypes, known for their different ability to generate embryogenic callus, were used as initial explants to establish callus cultures. Embryogenic and non-embryogenic calluses were obtained from the competent genotype (`Combi'), while only non-embryogenic callus was produced by the incompetent one (`Devon'). The morphogenetic competence of each callus type was evaluated by transferring some segments to regeneration conditions. The endogenous hormone concentrations (free indole-3-acetic acid [IAA], abscisic acid [ABA], gibberellins ₁, ₃ and ₂₀ [GAs], zeatin/zeatin riboside [Z/ZR] and N ⁶[²-isopentenyl] adenine/ N ⁶[²-isopentenyl] adenosine; [iP/iPA]) of the initial explants were determined by means of radio-immunoassay and showed that the only difference was the higher concentration of ABA found in the embryos of the most competent genotype; whose embryos showed a reduced rate of precocious germination. When analysing the endogenous hormone concentrations in the various callus types generated in each genotype, it was found that only differences in the free IAA concentrations were associated with variations in the morphogenic properties of the calluses. Higher concentrations of endogenous free IAA were typical of embryogenic callus cultures. It was also observed that a loss in the embryogenic competence of the calluses, due to a prolonged time of culture, occurred concomitantly with a reduction in free IAA concentrations, practically to the concentrations found in the non-embryogenic calluses.     

Effect of phytohormones on fiber initiation of cotton ovule

In order to study the effect of phytohormones on cotton fiber initiation, contents of four endogenous phytohormones and activities of four related enzymes in ovules (in vivo) of a fuzzless–lintless mutant (fl) and its wild-type (FL) line were measured from 4 days before anthesis (day −4) to 4 days after anthesis (day 4). The results showed that contents of indole-3-acetic acid, gibberellic acid (GA), and zeatin riboside in fl ovules were lower than those in FL ovules. Therefore, indole-3-acetic acid, GA, and zeatin riboside were thought to be the promoters of fiber initiation. Although abscisic acid (ABA) content in fl ovule was slightly higher than that in FL ovule on day 0, which might imply that ABA inhibited fiber initiation. Fiber initiation could also be influenced by enzyme through regulating synthesis and degradation of related phytohormones since fl ovules were significantly higher in activities of indole-3-acetic acid oxidase, cytokinin oxidase and peroxidase, but lower in activity of tryptophan synthetase than those in FL ovules. To test the above hypothesis, exogenous plant growth regulators were also applied for the culture of ovules from fl and FL in vitro. When no regulators were added, no fiber was induced on fl ovule, but a few fibers were induced in FL ovule. Higher total fiber units (TFU) were observed when indole-3-acetic acid and gibberellic acid (GA₃) were applied either separately or in combination to media. TFU did not increased with zeatin riboside alone, but the highest TFU was achieved when zeatin riboside was applied together with indole-3 acetic acid and GA₃, which implied that fiber initiation could be promoted by them as additive.     

Effects of elevated carbon dioxide and/or ozone on endogenous plant hormones in the leaves of <Emphasis Type="Italic">Ginkgo biloba</Emphasis>

Four-year-old Gingko (Ginkgo biloba L.) trees were exposed to ambient and elevated concentrations of CO₂ and O₃, and their combination for 1 year, using open-top chambers in Shenyang, China in 2006. Growth parameters and endogenous plant hormones were measured simultaneously over the experiment period. Elevated CO₂ increased leaf area and leaf dry weight but had no effect on shoot length, increased indole-3-acetic acid (IAA), gibberellins A₃ (GA₃), zeatin riboside (ZR), dihydrozeatin (DHZR) and isopentenyl-adenosine (iPA) content but decreased abscisic acid (ABA) content. Elevated O₃ significantly decreased leaf area, leaf dry weight, shoot length, and decreased IAA, GA₃, ZR content but increased ABA content and had a little effect on iPA, DHZR content. Elevated CO₂ + O₃ decreased IAA, iPA and DHZR content while increased ABA and GA₃ content in the early stage of the exposure and then decreased in the late stage. The evidence from this study indicates that elevated CO₂ ameliorated the effects of elevated ozone on tree growth, and elevated CO₂ may have a largely positive impact on forest tree growth while elevated O₃ will likely have a negative impact.     

Micropropagation,berberine content and antitumor activity of Jeffersonia dubia (Maxim.) Benth et Hook

An efficient micropropagation protocol was developed for Jeffersonia dubia using sucker explants. High frequency of multiple shoot formation was induced when the sucker explants were cultured on Chu’s (N6) medium with different concentrations of thidiazuron (TDZ) plus 0.54 µM α-naphthaleneacetic acid (NAA). The maximum frequency of shoot formation (96.2 %) was obtained on N6 medium with 2.27 µM TDZ plus 0.54 µM NAA. The highest mean number of shoots per explant (13.6) was obtained in temporary immersion system using an immersion frequency of 30 s every 30 min. The highest frequency of rooting (100 %), number of roots per shoot (5.8), and root length (6.3) was observed in half-strength N6 medium supplemented with 2.69 µM NAA. The regenerated plantlets (30 days old) were successfully acclimatized in the greenhouse with 98 % survival rate. The berberine content and cytotoxicity were higher in in vitro-developed calli and shoots than in leaves of field-grown plants. The greatest content of berberine was found in shoots (1381 μg g⁻¹) followed by calli (1092 μg g⁻¹) and leaves of field-grown plants (92 μg g⁻¹). At 1000 μg mL⁻¹ concentration, growth inhibition rate of berberine, callus, shoot, and leaf (in vivo) extracts were 68.4, 57.1, 54.2, and 17.7 %, respectively.

     

Melatonin influence on in vitro callus induction and phenolic compound production in sweet basil (Ocimum basilicum L.)

The aim of the present study was to evaluate melatonin effects on the callus induction and phenolic compound production of Ocimum basilicum L. (sweet basil). Calluses, derived from leaf explants, were grown on Murashige and Skoog (MS) medium supplemented with 0, 100, or 200 μM melatonin, and subsequently extracted for determination of their phenolic contents. Melatonin decreased the callus induction in both concentrations. Based on the phytochemical analysis, the highest total phenolic acid contents (784.6 μg g⁻¹ and 335.2 μg g⁻¹, respectively) were recorded in calluses grown in 100 and 200 μM melatonin-supplemented medium, compared with the calluses induced with MS alone (192.0 μg g⁻¹). Among the five phenolic acids confirmed in the callus samples, rosmarinic acid was the major constituent. The amount of rosmarinic acid increased significantly in callus grown on 100 μM melatonin medium by nearly 5-fold (754.2 μg g⁻¹), compared with the control group callus. Major volatiles in basil calluses were represented by 3-methylbutanal, benzaldehyde, 1,8-cineole, 2-nonenal, eugenol, and methyl eugenol, and these were in the ranges of 4 to 14%, 24 to 50%, 2 to 3%, 0 to 0.55%, and 2 to 17% (in relative percentages), respectively. The qualitative and quantitative analyses of these substances found in calluses formed on melatonin-supplemented or melatonin-free medium were evaluated separately.

     

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.     

γ-Irradiation of Barley Seeds and Its Effect on the Phytohormonal Status of Seedlings

We investigated the effect of γ-irradiation (4–50 Gy) of barley seeds (Hordeum vulgare L., cv. Nur) on the content of endogenous phytohormones–stimulators of plant growth and development: indol-3-acetic acid (IAA), indolyl-3-butyric acid (IBA), zeatin and abscisic acid (ABA). The ratio (IAA + IBA + zeatin)/ABA from the third to the seventh day of germination has been measured. It was shown that the changes in the content of phytohormones as a function of the radiation dose were nonlinear. In the dose range of 4–20 Gy, phytohormones balance was changed due to increased content of growth stimulators and decreased ABA content. Using a dose of 50 Gy led primarily to a decrease in the content of growth stimulators and an increase in ABA content, and the ratio (IAA + IBA + zeatin)/ABA shifted toward ABA content.