Effects of Sucrose Concentration on the Photosynthetic Ability of Rose Shoots In Vitro

Reducing the concentration of sucrose in the culture mediumover successive subcultures has been tested as a method forincreasing the ability of rose shoots grown in vitro (Rosa cvsIceberg and Peace) to take up CO₂. Shoots maintained on ‘constant’10, 20 and 40 g I^–1 sucrose showed decreased levels ofCO² uptake at higher sucrose concentrations, although cv. Peacegrew least at 10 g l^–1 and showed correspondingly lowamounts of CO₂ uptake compared with 20 and 40 g l^–1. Bothcultivars died when sucrose was omitted from the medium. Assucrose concentration was reduced in the medium, so CO₂ uptakeof shoots initially cultured on 20 and 40 g l^–1 sucrosewas found to increase, although a concentration of 10 gl ^–1sucrose seemed to be limiting, below which the growth and chlorophylllevels of shoots declined. Rosa hybrid, rose, shoot culture in vitro, photosynthetic ability, sucrose, infra-red gas analysis     

Photoperiodically Induced Changes in Seed Growth Rate of Soybean as Related to Endogenous Concentrations of ABA and Sucrose in Seed Tissues

Short-day photoperiods can increase the partitioning of assimilatesto filling seeds of soybean (Glycine max L. Merr.), resultingin higher seed growth rates. The plant growth substance ABAhas been implicated in the regulation of assimilate transferwithin filling soybean seeds. Thus, we hypothesized that anincreased concentration of endogenous ABA in seeds may enhancesucrose accumulation and seed growth rate of soybeans exposedto short-day photoperiods. Plants of cv. Hood 75 were grownin a greenhouse under an 8-h short-day photoperiod (SD) until11 d after anthesis (DAA) of the first flower, when half ofthe plants were transferred to a night-interruption (NI) treatment(3 h of low-intensity light inserted into the middle of thedark period). Plants remaining in SD throughout seed developmenthad seed growth rates 43% higher than that of plants shiftedto NI (7·6 mg seed^–1 d^–1 vs. 5·3 mgseed^–1 d^–1). On a tissue-water basis, the concentrationof ABA in SD seeds increased rapidly from 7.6 µmol l^–1at 11 DAA to 65·2 µmol l^–1 at 18 DAA, butthen declined to 6·6 µmol l^–1 by 39 DAA.In contrast, the concentration of ABA increased more slowlyin NI seeds, reaching only 47·4 µmol l^–1by 18 DAA, peaking at 57·0 µmol l^–1 on 25DAA, and declining to 10·2 µmol l^–1 by 39DAA. The concentration of sucrose in SD embryos peaked at 73·5mmol l^–1 on 25 DAA and remained relatively constant forthe remainder of the seed-filling period. In NI, the concentrationof sucrose reached only 38·3 mmol 1^–1 by 25 DAA,and peaked at 61·5 µmol l^–1 on 32 DAA. Thusin both SD and NI, sucrose accumulated in embryos only afterthe peak in ABA concentration, suggesting that ABA may havestimulated sucrose movement to the seeds. The earlier accumulationof ABA and sucrose in SD suggests that ABA may have increasedassimilate availability during the critical cell-division period,thus regulating cotyledon cell number and subsequent seed growthrate for the remainder of the seed-filling period. Glycine max L. Merr. cv. Hood 75, soybean, assimilate partitioning, abscisic acid, photoperiod, source-sink     

In Vitro Micropropagation of Delphinium malabaricum (Huth) Munz.--a Rare Species

A micropropagation technique was developed for Delphinium malabaricumusing nodes from inflorescence stalks Maximum shoot proliferationwas obtained on Murashige and Skoog's medium supplemented with2-1P (10 mg l^–1) and inositol (100 mg l^–1) Fromthe sixth passage onwards, shoots could be multiplied by omissionof inositol and reduction of 2-1P (0.5 mg l^–1) concentrationBest rooting response was obtained with a 24-h pulse treatmentof shoots with 0.5 mg I^–1 IBA in the dark, transfer oftreated shoots to hormone-free half-strength MS medium and incubationunder 24-h light. Regenerated plants were established successfullyin the field Cytological examination of root tips of in vitroand control plants showed identical chromosome number (2n =16) Delphinium malabaricum (Huth) Munz, micropropagation, tissue culture, rare plant     

The Uptake and Distribution of Copper in Some Forage Grasses as Affected by Nitrate-nitrogen Supply in Flowing Solution Culture

The effect of changes in nitrate-nitrogen supply on the absorptionand distribution of copper was examined in grasses grown inflowing solution culture with a maintained concentration ofcopper. Absorption by roots (µg Cu g^–1 dry root)decreased markedly when nitrogen had been depleted or was maintainedat 0.1 mg l^–1 N, but there was an immediate increase whennitrogen was maintained at 1.0 or 10.0 mg l^–1. There werealso large increases in the concentration of copper in the shootsof plants grown with 1.0 and 10.0 mg 1^–1 N. The rootsof plants grown with 0.1 or 1.0 mg 1^–1 N retained similarproportions of uptake, but a lower proportion was retained whenthe plants were grown with 10.0 mg 1^–1. Although a lowerproportion of the copper was associated with cell walls in theplants grown at 10.0 mg 1^–1 N this was the result of alower content of cell walls rather than an effect on copperitself. In a longer-term experiment in conventional solutionculture with a range of nitrogen concentration, the concentrationof copper in shoots was largely determined by shoot growth. Dactylis glomerata, Festuca arundinacea, Lolium perenne, cell walls, copper absorption, copper distribution, flowing solution culture, nitrate-nitrogen     

Callus Culture of Sainfoin (Onobrychis viciifolia) and Plant Regeneration through Somatic Embryogenesis

Callus of sainfoin (Onobrychis viciifolia Scop.) was initiatedfrom stem and root explants which were obtained from seedlingsgrowing in vitro, on Linsmaier Skoog (LS) medium supplementedwith 1 mg l^–1 2, 4-D and 1 mg l^–1 BA or only 1 mgl^–1 BA, and the Vacin and Went medium without hormones.Somatic embryos were formed on LS medium containing 1 m l^–1BA. Embryos developed into complete plants on filter paper saturatedwith hormone-free LS medium. Onobrychis viciifolia, somatic embryogenesis, callus culture, plant regeneration     

Micropropagation of Betula celtiberica

Plantlets were successfully regenerated from shoot segmentsof Betula celtiberica excised from young seedlings. Initiationand elongation of multiple shoot-buds were obtained after 20d culture in MS-modified medium plus BAP 0.6 mg l^–1 followedby 20 d culture in the same medium in the presence of a reducedBAP concentration (0.1 mg l^–1). Rooting was achieved 7d after having transplanted the isolated shoots to fresh medium,supplemented with IBA (0.2 mg l^–1). Betula celtiberica, birch, micropropagation, organogenesis     

The Growth and Development of the Wheat Apex: The Effects of Photoperiod on Spikelet Production and Sucrose Concentration in the Apex

Spring wheat (Triticum aestivum cv. Warimba) plants were grownin a controlled environment (20°C) in two photoperiods (8or 16 h). In the first instance, plants were maintained in eachof the photoperiods from germination onwards at the same irradiance(375 µE m^–2 s^–1). In the second case, allplants were grown in a long photoperiod until 4 days after double-ridgeinitiation when half the plants were transferred to a shortphotoperiod with double the irradiance (16 h photoperiod at225 or 8 h at 475 µE ^–2 s^–1). The rates of growth and development of the apices were promotedby the longer photoperiod in both experiments. Shoot dry weightgain was proportional to the total light energy received perday whereas the dry weight of the shoot apex increased withincreasing photoperiod even when the total daily irradiancewas constant. The principal soluble carbohydrate present in the shoot apexwas sucrose, although low concentrations of glucose and fructosewere found in the apices of long photoperiod plants late indevelopment. Sucrose concentration was invariably greater inthe slow-growing apices of short photoperiod plants, but roseto approach this level in the long photoperiod plants when theterminal spikelet had been initiated. Triticum aestivum, wheat, apex, spikelet initiation, photoperiod, flower initiation     

Somatic Embryogenesis from Clonal Leaf Tissues of Cassava

Leaf lobes were isolated from palmate leaves of clonal cassava(Manihot esculenta Crantz) material growing in vitro or in glasshouseconditions and subjected to a two-stage culture procedure involvingincubation on Murashige and Skoog (MS2) basal medium supplementedwith 2–12 mg l^–1 2,4-D for 20 d (Stage I) beforetransfer to MS2 basal medium supplemented with 0.01 mg l^–12,4-D and 0.1 mg l^–1 6-benzylamino purine (BAP) (StageII medium). Embryogenetic tissues, foliose structures and somatic embryosdeveloped from leaf lobes at all Stage I 2,4-D concentrations,except on those explants isolated from shoot-tip cultures incubatedon MS2 basal medium supplemented with 0.1 mg l^–1 NAA and1.0 mg l^–1 BAP. Leaf lobes isolated directly from glasshouse plants showed optimalembryogenetic competence when subjected to a Stage I cultureperiod of 17 d, although foliose structure initiation was optimalwith shorter Stage I durations. Leaf lobes of 2–4 mm lengthand those isolated from phyllotaxic leaf numbers 4 and 5 showedthe greatest embryogenetic competence. Manihot esculenta, cassava, somatic embryogenesis, tissue culture, morphogenetic competence     

Seed Water Relations and the Regulation of the Duration of Seed Growth in Soybean

Soybean [Glycine max (L.) Merrill] seeds and cotyledons weregrown in an in vitro culture system to investigate the relationshipsbetween cell expansion (net water uptake by the seed) and drymatter accumulation. Seeds or cotyledons grown in a completenutrient medium containing 200 mol m^–3 sucrose continueddry matter accumulation for up to 16 d after in planta seedsreached physiological maturity (maximum seed dry weight). Seedor cotyledon water content increased throughout the cultureperiod and the water concentration remained above 600 g kg^–1fresh weight. These data indicate that the cessation of seeddry matter accumulation is controlled by the physiological environmentof the seed and is not a pre-determined seed characteristic.Adding 600 mol m^–3 mannitol to the medium caused a decreasein seed water content and concentration. Seeds in this mediumstopped accumulating dry matter at a water concentration ofapproximately 550 g kg^–1. The data suggest that dry matteraccumulation by soybean seeds can continue only as long as thereis a net uptake of water to drive cell expansion. In the absenceof a net water uptake, continued dry matter accumulation causesdesiccation which triggers maturation. Key words: Glycine max (L.) Merrill, solution culture, duration of seed growth, water content, dry matter accumulation     

Influence of Genotype and Environment on Growth of Barley Embryos in vitro

Isolated embryos of three contrasting barley genotypes werecultured in vitro on a range of media comprising 16 combinationsof sucrose (3–12 per cent) and 2, 4-D (0–8 mg 1^–1)concentration. Cultures were incubated at a range of temperaturesfrom 5 to 25°C and were examined after 21 days when shootlengths as well as fresh and dry weights were recorded. Therelative influence on growth of increasing concentrations ofsucrose and 2, 4-D was investigated, as was their interactionwith the incubation temperature. The genotypes were found todiffer markedly in their response to these two media components,with each parameter of growth differentially affected. Resultsare discussed in relation to the known dormancy characteristicsof these genotypes. Hordeum vulgare L., barley, embryo, dormancy, 2, 4-dichlorophenoxy acetic acid, sucrose     

Biomass, feeding and production of Noctiluca scintillans in the Seto Inland Sea, Japan

The abundance and biomass of the large heterotrophic dinoflagellateNoctiluca scintillans, together with the changes in its potentialprey items, were monitored in the Seto Inland Sea, Japan, duringsummer 1997 (17 July-11 August). Growth and grazing rates ofNscintillans fed natural plankton populations were also measuredeight and seven times, respectively, during the survey period.The abundance and biomass of N scintillans averaged over thewater column (19 m) were in the range 1–345 cells 1^–1(temporalaverage = 93 cell1^–1) and 0.1–49.6 µg C l^–1(temporalaverage = 13.8 µg C l^–1; three times higher thanthat of calanoid copepods during the same period). Noctilucascintillans populations followed the changes in phytoplankton:N.scintillans biomass was increasing during the period of diatomblooms and was at a plateau or decreasing during periods oflow chlorophyll a. The growth rates of N.scintillans (µ)were also consistent with the wax and wane of the N.scintillanspopulation: N.scintillans showed highest growth rates duringdiatom blooms. A simple relationship between µ and chlorophylla concentration was established, and the production of N.scintillanswas estimated using this relationship and the measured biomass.The estimated production averaged over the water column wasin the range >0.1–5.2 µg C l^–1 day^–1(temporalaverage = 1.4 µg C l^–1 day^–1; 64% of the productionof calanoid copepods during the same period). Diatom clearancerates by N.scintillans were in the range 0.10–0.35 mlcell^–1 day^–1, and the phytoplankton population clearanceby N.scintillans was >12% day^–1. Thus, although thefeeding pressure of N.scintillans on phytoplankton standingstock was low, N.scintillans was an important member of themesozooplank-ton in terms of biomass and production in the SetoInland Sea during summer.     

Acclimation of Lolium temulentum to enhanced carbon dioxide concentration

Acclimation of single plants of Lolium temulentum to changing[CO₂] was studied on plants grown in controlled environmentsat 20°C with an 8 h photoperiod. In the first experimentplants were grown at 135 µ;mol m^–2 s^–1 photosyntheticphoton flux density (PPFD) at 415µl l^–1 or 550µll^–1 [CO₂] with some plants transferred from the lowerto the higher [CO₂] at emergence of leaf 4. In the second experimentplants were grown at 135 and 500 µmol m^–2 s^–1PPFD at 345 and 575 µl l^–1 [CO₂]. High [CO₂] during growth had little effect on stomatal density,total soluble proteins, chlorophyll a content, amount of Rubiscoor cytochrome f. However, increasing [CO₂] during measurementincreased photosynthetic rates, particularly in high light.Plants grown in the higher [CO₂] had greater leaf extension,leaf and plant growth rates in low but not in high light. Theresults are discussed in relation to the limitation of growthby sink capacity and the modifications in the plant which allowthe storage of extra assimilates at high [CO₂]. Key words: Lolium, carbon dioxide, photosynthesis, growth, stomatal density     

In Vitro Embryo Culture and Induction of Multiple Shoots in Lychee (Litchi chinensis Sonn.)

Immature embryos of different sizes and ages from commercialvarieties of lychee (Litchi chinensis Sonn.) were cultured ina range of different media. Embryos as small as 3 mm could becultured using in vitro techniques and subsequently grown intoplants. MS solid medium with 2% sucrose supplemented with 150ml l^–1 coconut water was most effective in stimulatingthe germination of immature lychee embryos. Embryos of lycheewere treated to induce adventitious buds from embryonic shootsas a means of achieving multiplication. The different varietiesexhibited differences in response, with Bengal embryonic shootsproducing 15 adventitious buds after pretreatment with 100 mgl^–1 BAP for 3 h. Root formation was achieved in 65% ofadventitious shoots using MS medium supplemented with 0.5 mgl^–1 NAA and activated charcoal. These plants were successfullydeflasked and grown on in the glasshouse. This technique providesof means of producing some multiple shoots from lychee embryosand has value for multiplication in a breeding program wherea method of micropropagation is unavailable. Litchi chinensis Sonn., lychee, embryo culture, multiple shoots, in vitro     

Multiplication In Vitro of Limonium estevei Fdez. Casas

Plantlets of Limonium estevei Fdez. Casas, an endangered Spanishspecies, were successfully regenerated from nodal segments excisedfrom young seedlings. Initiation of multiple adventitious budswere obtained in MS modified medium plus 1 mg l^–1 IBAand 0·1 mg l^–1 BAP. Rooting was achieved by transferof the isolated shoots to fresh MS medium without plant growthregulators. Fully grown plants were established in a pottingmix and are growing well in a greenhouse. Limonium estevei, in vitro multiplication, adventitious regeneration     

In Vitro Regeneration from Seedling Organs of Brassica oleracea var. italica Plenck cv. Green Comet I. Effect of Plant Growth Regulators

The calabrese cultivar Brassica oleracea var. italica cv. GreenComet was used in a study of the effects of exogenous hormoneson the growth and differentiation of seedling organs in vitro.Four types of explants were tested: hypocotyl segments, rootsegments, primary leaf discs and cotyledon discs. These explantswere incubated on media containing factorial combinations ofBAP x IBA, BAP x NAA, KN x IBA and KN x NAA (all at 0, 0.1,10 and 10.0mg l^–1). Hypocotyls were the most regenerativeexplants; shoot production was favoured by cytokinin: auxinratios greater than one and was decreased by IBA at 10 mg l^–1when callus was produced. Shoot formation from root explantsoccurred either in the absence of hormones or with low concentrations;no shoot was produced when any hormone was present at 10 mgl^–1. In contrast, shoot production from primary leaf diseswas favoured by high concentrations of both auxin and cytokininwith the combination of BAP and IBA the most effective. Shootproduction from cotyledon discs was sporadic with no consistentresponse on any auxin/cytokinin combination. After further experimentson the optimization of hormone concentration, the followingcombinations were chosen as allowing reliable regeneration:0.1 mg l^–1 BAP+0.1mg l^–1 IBA for hypocotyl segments,0.075 mg l^–1 KN +0.025 mg l^–1 IBA for root segments,and 5.0 mg l^–1 BAP+5.0 mg l^–1 IBA for leaf discs. Brassica oleracea var. italica, calabrese, tissue culture, seedling, auxin, cytokinin     

Somatic Embryogenesis and Plant Regeneration from Inflorescence Culture of Java Citronella: (Cymbopogon winterianus)

Embryogenic callus was induced from immature inflorescence segmentsof Java citronella (Cymbopogon winterianus) and maintained for2 years on Murashige and Skoog's medium supplemented with 2,4-D(l mg l^–1). The callus cells retained the original chromosomenumber of 2n = 20. The somatic embryos germinated into plantletson MS basal medium or medium with IAA, NAA, BAP or KN individually(l mg l^–1). The regenerated plantlets developed a goodroot system on full strength solid MS inorganics medium withIAA (1 mg l^–1). The regenerated plants were similar tothe donor plant in morphology and had the same chromosome number,but showed some variation in the essential oil content. Java citronella, Cymbopogon winterianus, somatic embryogenesis, regeneration, inflorescence culture     

Rates of ingestion and their variability between individual calanoid copepods: direct observations

The goals of this study were to determine rates of ingestionand fecal pellet release, and their variability, for individualplanktonic copepods over extended periods of time (>20 min).Ingestions and rejections of individual cells of the diatomThalassiosira eccentrica by adult females of the calanoid Paracalanusaculeatus were directly quantified by observing individual copepodscontinuously at cell concentrations ranging from 0.1 toli mm³l^–1.Average ingestion rates increased with increasing food concentration,but were not significantly different between 0.3 and 1.0 mm³l^–1(9.8 and 32.7 µg Cl^–1) of T.eccentrica. Rates ofcell rejections were low and similar at 0.1 and 0.3, but weresignificantly higher at 1.0 mm³ l^–1. The coefficientsof variation for average ingestion rates of individual copepodshardly differed between food concentrations, ranging from 17to 22%, and were close to those for average fecal pellet releaseintervals which ranged from 15 to 21%. A comparison betweenindividuals at each food concentration found no significantdifferences at 1.0; at 0.1 and 0.3 mm³l^–1, respectively,ingestion rates of four out of five females did not differ significantlyfrom each other. Average intervals between fecal pellet releaseswere similar at 0.3 and 1.0 mm³l^–1 of T.eccentrica, butsignificantly longer at 0.1 mm³ l^–1. Fecal pellet releaseintervals between individuals were significantly different ateach food concentration; these significant differences wereattributed to rather narrow ranges of pellet release intervalsof each individual female. Potential sources/causes of variabilityin the sizes and rates of copepods in the ocean are evaluated. ³Present address: Great Lakes Environmental Research Laboratory,2205 Commonwealth Boulevard, Ann Arbor, Ml 48105, USA     

Copper Nutrition of Sugarbeet

Sugarbeet (Beta vulgaris L.) plants were grown in refined sandat graded levels of copper ranging from acute deficiency (0.000325µg Cu cm^–3) to excess (65 µg Cu cm^–3).Visible effects of copper deficiency appeared up to 0-00065µg Cu cm^–3and depression in growth up to 00065µCucm^–3. Copper deficiency decreased the concentrations ofDNA and RNA and the activities of polyphenol oxidase, cytochrome-coxidase, catalase and aldolase; and it increased the activitiesof peroxidase, ribonuclease and acid phosphatase in leaves.The maximum sucrose concentration in roots was obtained at 0-65µCucm^–3 Twenty four h after infiltration of a solution of 65µCucm^–3into copper deficient leaves, the activities of cytochrome-coxidase and peroxidase had increased even in the presence ofcycloheximide but that of polyphenol oxidase increased onlyin the absence of this inhibitor. Key words: Beta vulgaris, Cu deficiency: Enzymes     

Soyabean Stem In Vivo Nitrate Reductase Activity

Stem from three- and four-week-old Soyabean [Glycine max (L.)Merr. cv. Tracy] plants reduced from 0.3 to 0.7 µmol nitrateh^–l g^–l f. wt. Leaf activity was 4.7–7.6 µmolnitrate h^–l g^–l f. wt. Outer stem was two to fourtimes more active at reducing nitrate than was inner stem. Plantnitrate nutrition had a strong effect upon the ratio of activitypresent in stem and leaf. More nitrate increased the proportionpresent in leaves. Glycine max L., soyabean, nitrate assimilation, nitrogen metabolism, Rhizobium japonicum     

Somatic Embryogenesis and Its Hormonal Regulation in Tissue Cultures of Freesia refracta

Somatic embryogenesis can be induced in tissue cultures of Freesiarefracta either directly from the epidermal cells of explants,or indirectly via intervening callus. These two pathways ofsomatic embryogenesis can be controlled and regulated by varyingthe combinations and levels of exogenous hormones. When younginflorescence segments were cultured in vitro on modified N₄(MN₄) medium supplemented with 2 mg l^–1 indoleacetic acid(IAA) and 3 mg l^–1 6-benzylaminopurine (BAP), some ofthe epidermal cells began to exhibit the features of embryogeniccells. These cells produced embryoids and developed into newplants through direct somatic embryogenesis. If the same explantswere placed on Murashige and Skoog's (MS) medium containing2 mg l^–1 IAA, 05 mg l^–1 BAP and 05 mg l^–1naphthaleneacetic acid (NAA), pale-yellow translucent nodularcalluses appeared on the surface of the explants. When thiskind of callus was transferred to MN6 medium with 2 mg l^–1IAA and 3 mg l^–1 BAP, embryoids formed which further developedinto plantlets. The regenerated plants were morphologicallynormal and possessed the normal diploid chromosome number of2n = 22. A similar result has also been obtained with youngleaf explants of this plant. The early segmentations of embryogeniccells and the development of embryoids were studied using histologicaland scanning electron microscopic techniques, and the resultshave been discussed in association with the ontogeny and originof the embryoids. Freesia refracta Klatt, somatic embryogenesis, plant regeneration, exogenous hormones