Showing papers on "Zeatin published in 1987"

Plant regeneration in vitro from embryogenic cultures of spring- and winter-type barley (Hordeum vulgare L.) varieties.

Abstract: Immature embryos of 41 lines of barley were screened in vitro for callus induction and somatic embryogenesis on different media to establish totipotent cultures. The use of modified MS and CC media, both supplemented with 1 g/l casein hydrolysate, and the substitution of agarose for agar resulted in the highest frequencies of somatic embryo induction. Embryogenic callus was induced and plants regenerated from 23 of the lines tested. The auxins 2,4-D, dicamba, picloram and 2,4,5-T were suitable for embryogenic callus induction. High frequencies of somatic embryo germination occurred on CC medium supplemented with 1 mg/l IAA and 0.05 mg/l zeatin. A strong genotypic effect on the capacity and frequency of embryogenic callus formation was found. Cultivar Golden Promise always gave the best results. Experiments with field grown material in 3 consecutive years showed that environmental factors also strongly influenced the induction of somatic embryogenesis and plant regeneration.

The Physiological Basis for Cytokinin Induced Increases in Pod Set in IX93-100 Soybeans.

Abstract: Previous investigations have shown the feasibility of increasing pod number on legumes by the application of 6-benzylaminopurine (BA) directly to the raceme. These investigations were designed to determine what reproductive parameter was affected by cytokinin application, and if these applications were overcoming a deficiency in root-produced cytokinins during late flowering. Five individual main stem racemes on greenhouse grown soybeans (Glycine max L. Merr.) were treated with 2 millimolar BA. A single application of BA when pods appeared at 25 to 50% of the proximal floral positions resulted in a 58% increase in pod set due primarily to a 33% reduction in floral abscission. Applications of BA at later intervals also resulted in significant reductions in total abscission. When three applications of BA were imposed on the upper five nodes of field grown soybeans, total pod number and seed weight were significantly increased in this section of the canopy by 27 and 18%, respectively. Throughout the flowering period, root pressure exudate was sampled for the subsequent separation and quantification of zeatin, dihydrozeatin, zeatin riboside, dihydrozeatin riboside, and isopentenyladenine. Total cytokinin flux peaked from 0 to 9 days after flowering began, and then dropped to one-half of this level by 15 days postanthesis. The probability that a flower would initiate a pod was directly related to the concentration of total cytokinins present in the exudate when the flower opened.

Plant Growth Regulators and Morphogenesis in Cell and Tissue Culture of Forest Trees

Abstract: Plant growth regulators include naturally occurring plant hormones such as indoleacetie acid (IAA), gibberellins, zeatin, abscisic acid (ABA) and ethylene, etc., and also a number of synthetic chemicals that affect or control growth and development in plants. Each type of plant growth regulator has a wide range of physiological effects in different plants. These effects are determined by the kind of the growth regulator, its concentration, the presence or absence of other growth regulators, and by the genetic makeup and the physiological status of the target tissue. The same physiological response in different tissues even of the same plant may require different growth regulator(s) or different combinations of growth regulators. Synergism and quantitative interaction of two or more growth regulators are of common occurrence. Finally, a growth regulator that elicits a positive response in a given tissue at a given concentration may inhibit the same physiological response when used at higher concentrations.

Cytokinin translocation and metabolism in lupin species. I. Zeatin riboside introduced into the xylem at the base of Lupinus angustifolius stems.

Abstract: The principal cytokinins in xylem exudate of blue lupin (Lupinus angustifolius L.) identified by radioimmunoassay and mass spectrometry were zeatin riboside (ZR), dihydrozeatin riboside and zeatin, but cytokinin O-glucosides and nucleotides were also present. Radioactive [3H]ZR was supplied to the transpiration stream of both derooted and intact blue lupin plants. The distribution of methanol-extracted radioactivity throughout the shoot and the identities of 3H-labelled metabolites were determined; for the latter work, a new bilayer thin-layer chromatography system was developed. In derooted plants supplied with ZR for about 20 h, radioactivity per unit fresh weight of tissue was greatest in the peduncle, stem and axillary shoots and least in the seed. Differences in metabolism were more marked between tissue types than within tissues of different maturity. The principal metabolites in derooted plants (ZR supplied for up to 24 h) were: in laminae, O-glucosyldihydrozeatin, lupinic acid, dihydrozeatin riboside and adenosine; in petioles, dihydrozeatin riboside and adenosine; in pod walls, cytokinin nucleotides, dihydrozeatin riboside, adenine and adenosine; in developing lateral shoots, cytokinin nucleotides; in stem, dihydrozeatin riboside, dihydrozeatin and cytokinin nucleotides. Apart from adenosine, no metabolites could be identified with certainty in seed. In experiments with intact plants conducted over a longer period (plants extracted 2 days after 30 h uptake), metabolism was more extensive but, in all tissues except seed, low percentages of radioactivity were still due to metabolites with an intact zeatin moiety (most frequently O-glucosyldihydrozeatin and lupinic acid). Free zeatin and dihydrozeatin were not found in any tissue and their ribosides were detected with certainty only in pod walls (<2% of extracted 3H). Extraction with 0.5 N KOH after extraction with methanol yielded, on average, an additional equal amount of radioactivity due to nucleotides of adenine and guanine. Features of the translocation studies include: (1) the direct lateral movement of ZR and/or related cytokinins from xylem to bark which was established by bark ringing experiments; (2) the relatively high level of radioactivity per unit tissue fresh weight in developing lateral shoots; (3) the lack of appreciable movement to the seed although the cytokinin reached the pod walls and was partially conserved there; (4) the disproportionately high retention of radioactivity by the petioles which increased with leaf age; (5) changes in ZR metabolites in laminae associated with age; (6) the detection of a new nucleotide metabolite of ZR in pod walls, bark and lateral shoots; this appears to be a transient metabolite with an intact ZR moiety and may be involved in ZR uptake and/or transport. The studies of ZR translocation and metabolism are discussed in relation to sequential leaf senescence, the proposal that developing seeds accumulate xylem cytokinins, and the origin of phloem cytokinins.

Plant regeneration from cultured immature embryos of Sorghum bicolor (L.) Moench.

Abstract: Immature embryos of 20 sorghum genotypes were cultured on MS 5 medium containing MS mineral salts supplemented with 2,4-D, zeatin, glycine, niacinamide, Ca-pantothenate, L-asparagine, and vitamins. For regeneration, calli were transferred onto the same medium with the exception that IAA was substituted for 2,4-D. In general, immature embryos obtained 9–12 days after pollination resulted in the best redifferentiation. Ability of calli to regenerate varied among genotypes; cultivars C401-1 and C625 had the highest redifferentiation frequencies. Ability to redifferentiate was heritable and acted as a dominant trait. At least two gene pairs were involved. Regenerated R0 plants were planted in a greenhouse and their selfed (R1 and R2) progenies were planted in the field and examined for morphological and cytological variations. The majority of the phenotypic variations noted in R0 were not transmitted to later generations. However, variants for plant height, degree of fertility, and midrib color persisted in R1 and R2 generations. A variation in tallness was attributable to one dominant mutant gene. Short stature and male sterility variants appeared to be consequences of recessive mutant genes controlling those traits. Minor variations in peroxidase banding patterns were found among R0 plants.

Plant regeneration from protoplast culture of sweet potato (Ipomoea batatas Lam.).

Abstract: This is the first report on successful plant regeneration from protoplasts of sweet potato. Two cultivars (Guyana and Duclos XI) of sweet potato plants propagated under in vitro conditions were used as the source of protoplasts. Green compact calli with meristematic areas were induced in the medium supplemented with 2mg1−1 zeatin, and plant regeneration occurred when these calli were transferred onto the medium with zeatin level reduced to 0.25mg1−1. Plant regeneration was found to be genotype-dependent, since it was only obtained for cultivar Duclos XI.

Isolation and partial purification of an enzyme catalyzing the formation of O-xylosylzeatin in Phaseolus vulgaris embryos.

Abstract: An enzyme catalyzing the formation of a cytokinin metabolite, an O-pentosyl derivative of zeatin [Lee, Y. H., Mok, M. C., Mok, D. W. S., Griffin, D. A. & Shaw, G. (1985) Plant Physiol. 77, 635-641], was isolated from Phaseolus vulgaris embryos. Of all the potential pentose donors tested, UDP-xylose was the only substrate recognized by the enzyme. This indicates that the O-pentosyl derivatives previously obtained are O-xylosylzeatin and its ribonucleoside. The enzyme (UDP-xylose:zeatin O-xylosyltransferase, EC 2.4.2.-) has high affinity for trans-zeatin (Km 2 μM) and dihydrozeatin (Km 10 μM) but does not recognize cis-zeatin or ribosylzeatin. The molecular weight of the enzyme is approximately 50,000 and the pH optimum of the reaction is 8-8.5. Under comparable isolation and reaction conditions, similar enzyme activity could not be detected in P. lunatus embryos, confirming the genetic differences observed in vivo.

Phytohormones,Rhizobium mutants, and nodulation in legumes. VII. Identification and quantification of cytokinins in effective and ineffective pea root nodules using radioimmunoassay

Abstract: Radioimmunoassays (RIA), employing antisera raised in rabbits against bovine serum albumin conjugates of zeatin riboside, dihydrozeatin riboside, and isopentenyladenosine, were used to estimate levels of these cytokinins and their corresponding bases in samples of effective (nitrogen-fixing, Fix+), ineffective (nonnitrogen-fixing, Fix−) pea root nodules and uninoculated roots. Assays were done on extracts of nodule tissue, 1–2 g fresh weight, or approximately 10 g fresh weight of root tissue, and high specific activity [3H]zeatin riboside was added during preparation of the extract for use as a recovery marker. Two different purification procedures were employed, each involving several purification steps. High performance liquid chromatography (HPLC) was the final step in both procedures. Fractions from HPLC were analyzed by RIA using the appropriate antiserum. The cytokinins, zeatin, zeatin riboside, dihydrozeatin riboside, isopentenyl adenine, and isopentenyladenosine were detected and quantified in nodule tissue, and similarly, in root tissue (with the exception of zeatin, which we were unable to quantify in root tissue). Cytokinin levels in nodule tissue were higher than those in root tissue. The major cytokinins detected in nodule tissue were zeatin, followed by zeatin riboside and then dihydrozeatin riboside. The levels of zeatin and zeatin riboside estimated in nodules in the present study by RIA were of the same order of magnitude, though tending to be a little higher, than values obtained previously by bioassay. Dihydrozeatin riboside was identified with confidence for the first time in nodule tissue. There was a general decline with age in cytokinin levels in nodules, but no major qualitative change in nodule cytokinins with age. For theRhizobium strains examined, the data did not indicate a clear correlation between nodule cytokinin levels and the effectiveness of nodules in nitrogen fixation.

Plant regeneration from stem cortex protoplasts of Brassica napus

Abstract: Cell layer strips composed of the epidermis and 7–9 layers of subepidermal cells were isolated from the 3–4 terminal internodes of Brassica napus cv Westar plants at the early flowering stage. The strips were precultured for one day in modified liquid MS [11] medium and subsequently incubated for 17–18 h in a 0.4 M mannitol solution containing 1% Macerozyme and 1% Cellulase ‘Onozuka’ R-10. Protoplast yield was 2–2.8×106 per 1.0g of tissue. Protoplasts were cultured at 1×105/ml in three different media: S1 [13], B [12] and L[8]. The first cell divisions occurred after 2–8 days of culture at frequencies of 20–54%. The highest growth rate of colonies was obtained in L medium containing 0.4 M sucrose and 2% Ficoll. After 4 weeks, green calli, 1–2 mm in diameter were transferred onto B5 [2] medium with 3 mgl-1 zeatin, 1% sucrose, 0.1 M mannitol and 0.5% agarose for shoot regeneration. Up to 20% of the calli regenerated shoots which subsequently were rooted and established in soil in the greenhouse.

Cytokinins in cut carnation flowers. II. Relationship between endogenous ethylene and cytokinin levels in the petals

Abstract: Tentative identification using HPLC and RIA techniques indicated the presence of zeatin-O-glucoside, zeatin, ribosylzeatin, dihydrozeatin, iso-pentenyladenine and iso-pentenyladenosine in the petals of carnation flowers. Dihydrozeatin is apparently responsible for most of the biological activity. Within the petals most activity was detected in the basal parts which also senesced much slower than the upper parts of the petals. Treatment with AOA extended petal longevity and reduced ethylene production. This was associated with higher cytokinin-like activity in the basal parts of the petals.

Use of isopentenyladenosine and dihydrozeatin riboside antibodies for the quantification of cytokinins

Abstract: Antisera have been raised in rabbits against dihydrozeatin riboside and isopentenyladenosine, and their cross-reactivity characteristics have been examined in detail. These antisera, together with an antiserum previously raised against zeatin riboside, have been employed in radioimmunoassays. Separative procedures that enable a wide range of naturally occurring cytokinins to be separated prior to analysis by radioimmunoassay have been developed. The accuracy with which the following cytokinins can be quantified by our methods, which employ tritiated cytokinin recovery markers, has been estimated: zeatin riboside, zeatin, dihydrozeatin riboside, dihydrozeatin, O-glucosyl zeatin riboside, O-glucosyl zeatin, O-glucosyl dihydrozeatin riboside, O-glucosyl dihydrozeatin, zeatin-9-glucoside, zeatin-7-glucoside, lupinic acid, isopentenyladenosine, and isopentenyladenine.

Effects of growth regulators on light‐dependent anthocyanin production in Zea mays seedlings

Abstract: Rengel, Z. and Kordan, H. A. 1987. Effects of growth regulators on light-dependent anthocyanin production in Zea mays seedlings. The effects of ethylene, indolyl- and naphthylacetic acids, zeatin, benzyladenine, gib-berellic acid and triiodobenzoic acid on anthocyanin production in seedlings of Zea mays L. cv. Golden Bantam were investigated. Endogenously produced and exogen-ously supplied ethylene, as well as the other growth regulators tested markedly suppressed anthocyanin formation. Except for triiodobenzoic acid, the other growth regulators stimulated ethylene production, the amounts produced in the light being larger than those in the dark. Absorption of ethylene by permanganate as well as inhibition of ethylene production or action by Co2+ or Ag+ increased anthocyanin formation in maize seedlings above the level found in the control plants. The inhibiting effect of auxins and cytokinins on anthocyanin production was reversed by Co2+ or Ag+. In contrast, decreased anthocyanin formation caused by gibberellic acid or triiodobenzoic acid seemed unrelated to ethylene and could not be alleviated by Co2+ or Ag+.

Plant regeneration from Actinidia callus cultures

Abstract: SummaryCallus was initiated in vitro from small portions of bark with cambial tissue of kiwi. Optimum regeneration was achieved with Gamborg B5 medium containing BA, compared with 2iP and zeatin. Some of the shoots obtained were grown into complete plants and acclimatized in the greenhouse.

Cultural behavior of protoplasts from different organs of eggplant (Solanum melongena L.), and plant regeneration

Abstract: Plants of Solanum melongena were propagated under in vitro conditions (27°C, 12h/day illumination at 62 μEm-2s-1, 60% humidity) by subculture of terminal and lateral cuttings on MS medium +20 gl-1 sucrose + Morel and Wetmore vitamins at 1/8 strength and 7 gl-1 agar. Lamina, petioles and stems of 3-week-old cuttings were used as sources of protoplasts. The best mean yield of protoplasts was obtained from the lamina with 9,030×103 protoplasts per gram of tissue. Petioles and stems yielded respectively 3,144×103 and 1,220.4×103 protoplasts per gram of tissue. first division of petiole and stem protoplasts occurred within 48 h, while lamina protoplasts underwent division after 3–4 days of culture in KM8p medium +2,4-D(0.2 gl-1) + zeatin (0.5 mgl-1) + NAA (1 mgl-1) and 0.35M glucose as osmoticum. The highest percentage of dividing cells was obtained from petiole material, estimated at 33.4% after 7 days, compared to 23.8% and 19.4% respectively for stem and lamina protoplasts. When BAP replaced zeatin in KM8p, the division percentage of lamina protoplasts was reduced to 10–15%. When transferred to regeneration medium, all calli derived from KM8p + zeatin formed deep-green spots identified as embryo-like structures, while only few calli from KM8p + BAP underwent shoot organogenesis without formation of green spots. Some of embryo-like structure developed into plantlets with a frequency of 1–2 plantlets per callus especially on MS medium + zeatin (4 mgl-1) + IAA (0.2 mgl-1). Maintaining protoplast-derived calli on MS + BAP (0.5 mgl-1) + NAA (0.5 mgl-1) for more than 3 weeks resulted in a decrease and loss of cell totipotency.

Dynamics of production of trans -zeatin and trans -zeatin riboside by immobilized cytokinin-autonomous and cytokinin-dependent tobacco cells

Abstract: Two strains of cultured tobacco cells (Nicotiana tabacum L. cv. Wisconsin 38) differing in their requirement for exogenous cytokinins (cytokinin-dependent and cytokinin-autonomous) were immobilized on polyphenylenoxide (Sorfix) activated with glutaraldehyde. Columns packed with immobilized cells were continually eluted with diluted Murashige and Skoog's medium lacking or supplemented with synthetic cytokinin (6-benzylaminopurine; BA). Purified samples of column eluates were fractionated by HPLC, andtrans-zeatin (t-Z) andtrans-zeatin riboside (t-ZR) content was estimated by enzyme immunoassay. Both cytokinin-autonomous and cytokinin-dependent tobacco cells produced and excretedt-Z and its riboside, and there were significant quantitative differences between the strains. The steady-state excretion rate oft-Z was 19.8 ng · g−1 dw · h−1 and 4 ng · g−1 dw · h−1, respectively, and that oft-ZR 4 ng · g−1 dw · h−1 and 1 ng · g−1 dw · h−1, respectively. Exposure of cytokinin-dependent cells to BA after 72 h of starving for this synthetic cytokinin caused temporary increase in excretion of both zeatin and its riboside. After the application of 5 μM BA for 24 h, the excretion rate oft-ZR reached 5 ng · g−1 dw · h−1 (5-fold increase), and that oft-Z achieved 12 ng · g−1 dw · h−1 (3-fold increase). The elevation oft-Z excretion was delayed about 13 h compared witht-ZR excretion, which started increasing almost immediately after BA application. A pulse of BA in lower concentration (1.5 μM for 30 h) provoked lower response.

Cytokinins in cut carnation flowers: I. The complex in ovaries

Abstract: Tentative identification of the cytokinins present in extracts of Dianthus caryophyllus ovaries using High Performance Liquid Chromatography and Radioimmunoassay techniques, revealed the presence of trans-ribosylzeatin, trans-zeatin, dihydrozeatin and N6 (△2-isopentenyl)adenine. In addition slow moving compounds (paper chromatography) which could be hydrolysed by β-glucosidase were also detected. After hydrolysis the active compounds co-chromatographed with zeatin and ribosylzeatin.

In vitro regeneration of apomictic bluestem grasses

Abstract: Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes ≤8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity.

Determination by Gas Chromatography-Mass Spectrometry of [15N5]Adenine Incorporation into Endogenous Cytokinins and the Effect of Tissue Age on Cytokinin Biosynthesis in Datura innoxia Crown Gall Tissue

Abstract: In this study gas chromatographic-mass spectrometric techniques have been used to identify and quantify the metabolic incorporation of [15N5]adenine into zeatin and its metabolites by 3-week-old Datura innoxia Mill, crown gall tissue. In a parallel study the levels of endogenous cytokinins were also determined by the stable isotope dilution technique using deuterium (2H)-labeled internal standards. Incorporation levels of the [15N5]adenine after 8 hours of incubation, expressed as a percentage of the endogenous cytokinins, were as follows: zeatin (1.0%), zeatin riboside (1.5%), and zeatin riboside 5′-phosphate (10.2%). These results are consistent with those observed in complementary experiments using [U-14C]adenine, and support the proposal that the cytokinin biosynthesis occurs primarily at the nucleotide level. The effect of tissue age on cytokinin biosynthesis, determined by [U-14C]adenine incorporation into cytokinins by tissues at varying growth stages, indicated a steady increase with time reaching maximal synthesis at five weeks following subculture after which the level of 14C incorporation into cytokinins declined.

Metabolism of 14C-Zeatin in Phaseolus Embryos : Occurrence of O-Xylosyldihydrozeatin and Its Ribonucleoside

Abstract: The metabolism of trans-[8-14C]zeatin was examined in embryos of Phaseolus acutifolius A. Gray P.I. 321637 and Phaseolus coccineus Lam. cvs Scarlet Runner and Desiree. In both species zeatin was converted to ribosylzeatin, ribosylzeatin 5′-monophosphate, O-glucosyl-9-ribosylzeatin and the recently discovered O-xylosyl derivatives of zeatin and ribosylzeatin (Turner, JE, DWS Mok, MC Mok, G Shaw 1987 Proc Natl Acad Sci USA. In press). Two new metabolites, identified by enzyme degradation and gas chromatography-mass spectrography analyses as O-xylosyldihydrozeatin and its ribonucleoside, were recovered from P. coccineus embryos. From this and previous studies it may be concluded that the potential to form O-xylosyl derivatives of zeatin is present only in embryos of three Phaseolus species (P. vulgaris L., P. coccineus, and P. acutifolius), but not in P. lunatus L., while the reduction of the side chain is most prominent in P. coccineus.

Plant regeneration from leaf protoplasts of Solanum torvum

Abstract: A protocol to obtain regenerated plants from protoplasts of Solanum torvum Sw a wild species of eggplant resistant to Verticillium wilt is reported. Leaf protoplasts were enzymatically isolated from six-week old seedlings grown in a controlled environment chamber. Protoplasts were plated on modified KM medium (0.4 M glucose)+(mg/l): 1.0 p-chlorophenoxyacetic acid (CPA)+1.0 naphthaleneacetic acid (NAA)+0.5 6-benzylaminopurine (BAP) and 0.02 abscisic acid (ABA). The protoplast density was 5×104 per ml with 5 ml placed in each of two quadrants in X-dishes (100×15 mm). The reservoir medium was modified KM+(mg/l): 0.1 NAA+0.5 BAP+0.1 M sucrose+0.1 M mannitol+0.6% washed agar+1% activated charcoal. Dishes were initially placed in the dark at 27°C. Protoplast division was initiated in 1–2 weeks and 4 weeks later p-calli were 1–3 mm. Plating efficiency was 11% when measured at 3 weeks. Six-week old p-calli were transferred individually onto Whatman No. 1 filter paper layered on modified KM (0.15 M sucrose)+mg/l: 2.0 indoleacetic acid (IAA)+2.0 zeatin+0.5% washed agar for 2 weeks. Subsequently, shoots occurred within 4 weeks at 70% efficiency on MS+30 g/l sucrose+2 mg/l zeatin. Shoots were rooted on half strength MS+10 g/l sucrose.

Plant regeneration by pollen embryogenesis from cultured whole spikes of barley (Hordeum vulgare).

Abstract: Pollen embryogenesis and subsequent plant regeneration have been established from cultured whole barley spikes in agitated N6 liquid medium (Chu 1978) containing high levels of 2,4-D, Ficoll and potato extract. Microspore division within the anthers and subsequent embryogenic development were obtained in medium containing high amounts of reduced nitrogen with Zeatin, NAA and BAP (all at 0.5 mg/l levels, pH 6.2). Once embryoids were formed in the liquid medium, they produced secondary embryoids from the scutellum and subsequently plants on MS (Murashige and Skoog 1962) agar medium containing BAP and NAA. The ratio of green plants to albino was 1∶8.7.

Hydroxylation of N6-(Δ2-Isopentenyl)adenine to Zeatin Relative Activities of Organ Systems from Actinidia arguta

Abstract: By incubating explants from Actinidia arguta seedlings on a nutrient medium supplemented with 20 to 30 micromolar N(6)-(Delta(2)-isopentenyl)adenine (i(6)Ade) and then measuring zeatin (io(6)Ade) accumulation in tissues, the distribution of i(6)Ade hydroxylase activities in whole plants could be determined. Based on analyses with three entire plants, it is estimated that, as an organ system, roots contain approximately 68% of the plant's hydroxylase, while stems and leaves account for about 26% and 6%, respectively, of the total activity. Depending on the part of the root examined, hydroxylase activities ranged from 20 to 148 nanomoles io(6)Ade accumulated per gram fresh weight per 24 hours of incubation. Stem activities ranged from 17 to 165 nanomoles per gram fresh weight per 24 hours with the lowest activities being found at the tip. Leaf activities were substantially lower (1-10 nanomoles per leaf depending on position) than either root or stem.

Plant regeneration from seedling explants of perennial Glycine species

Abstract: More than 5000 cultures, from 30 accessions of six Glycine species, were established to assess the role of plant genotype in the response to an agar-solidified culture medium containing B5 salts and vitamins, 3% w/v sucrose, 1.1 mg 1−1 BAP and 0.005 mg 1−1 IBA, already known to induce shoot regeneration in callus of G. clandestina. Shoot initiation was obtained in a variety of explants from G. canescens, G. falcata, G. latrobeana and G. tomentella. With the exception of G. latrobeana, development of buds into shoots followed transfer to B5-based medium with 0.2 mg−1 BAP and 0.005 mg 1−1 IBA. Shoots readily produced roots in hormone-free half-strength B5 medium. In G. latrobeana, both extension and rooting occurred on this medium. Shoot regeneration was obtained in 12 of 30 accessions evaluated, but one accession of G. canescens, G1171, produced shoots and plantlets at a consistently higher frequency than other accessions, with plantlet recovery in more than 70% of the cultures. Bud formation in callus of G. canescens G1171 also occurred if BAP was replaced by 1.0 mg 1−1 kinetin, 2i-p or zeatin, albeit at a lower frequency.