Isolation and culture of protoplasts from mesophyll tissue of the legume Cyamopsis tetragonoloba L.

Protoplasts of Cyamopsis tetragonoloba were isolated from leaves of in vitro grown plants. The yield of the protoplasts, their viability and subsequent divisions were greatly influenced by the pH of the media used for isolation and culture of protoplasts. Sustained divisions of the cultured protoplasts were best supported by a modified Kao and Michayluk (1975) nutrient medium containing glucose (0.4 M), NAA (4 mgl^–1), 2,4-D (1 mgl^–1) and KIN (2 mgl^–1 ). The protoplast derived cells developed calli on transfer to Murashige and Skoog (1962) medium supplemented with 1 mgl^–1 each of 2,4-D, NAA and KIN.     

Micropropagation of sugarcane (Saccharum spp.)

A method for callus induction, adventitious bud regeneration, shoot multiplication and rooting of in vitro formed shoots of Helianthus annuus L. var. Argentario is described. Hypocotyl and cotyledon explants formed callus on medium containing 2 mgl^–1 naphthalene acetic acid and 0.5 mgl^–1 benzyladenine. Adventitious buds were formed on hypocotyl segments on medium containing 0.5–2 mgl^–1 benzyladenine. The optimal level of sucrose concentration for shoot regeneration from hypocotyls was 1.5%. Multiplication from shoot apices was promoted by kinetin (2 mgl^–1) plus gibberellic acid (5 mgl^–1), benzyladenine (2 mgl^–1) plus gibberellic acid (10 mgl^–1) or at lower frequency by benzyladenine (1 mgl^–1). A general feature of the plantlets formed in vitro was the precocious flowering.     

Internal loading of nutrients and certain metals in the shallow eutrophic Lake Myvatn, Iceland

Sub-arctic Lake Myvatn is one of the most productive lakes in the Northern Hemisphere, despite an ice cover of 190 days per year. In situ, transparent and dark flux chambers were used for direct measurements of benthic fluxes of dissolved oxygen, nutrients, silica and certain metals, taking into account primary production and mineral precipitation. The range of benthic flux observed for dissolved oxygen (DO), dissolved inorganic carbon (DIC), ammonium, ortho-P, silica, calcium, and magnesium was –45.89 to 187.03, –99.32 to 50.96, –1.30 to 1.27, –0.51 to 0.39, –62.3 to 9.3, –33.82 to 16.83, and –23.93 to 7.52 mmol m^–2 d^–1, respectively (negative value indicating flux towards the lake bottom). Low benthic NH₄ ⁺ and ortho-P fluxes were likely related to benthic algal production, and aerobic bottom water. Ortho-P fluxes could also be controlled by the dissolution/precipitation of ferrihydrite, calcite, and perhaps hydroxyapatite. The negative silica fluxes were caused by diatom frustule synthesis. Benthic calcium and magnesium fluxes could be related to algal production and dissolution/precipitation of calcium and/or Ca,Mg-carbonates. Fluxes of DO, DIC, pH and alkalinity were related to benthic biological processes. It is likely that some of the carbon precipitates as calcite at the high pH in the summer and dissolves at neutral pH in the winter. Mean of the ratio of gross benthic DIC consumption and gross benthic DO production was 0.94 ± 0.18, consistent with algal production using NH₄ ⁺ as N source. During the summer weeks the water column pH remains above 10. This high pH is caused by direct and indirect utilisation of CO₂, HCO₃ ^–, CO₃ ^–2, H₄SiO₄ ^° and H₃SiO₄ by primary producers. This study shows that in shallow lakes at high latitudes, where summer days are long and the primary production is mostly by diatoms, the pH is forced to very high values. The high pH could lead to a positive feedback for the Si flux, but negative feedback for the NH₄ ⁺ flux.     

Micropropagation of Capparis decidua (Forsk.) Edgew. — a tree of arid horticulture

A method for micropropagation of mature trees of Capparis decidua was developed. Multiple shoots were obtained from nodal explants on Murashige and Skoog's (1962) medium+0.1mgl^–1 NAA+5.0mgl^–1BAP+additives (50mgl^–1 ascorbic acid and25 mgl^–1 each of adenine sulphate, L-arginine and citric acid) at 28 ± 2°C, 12 h/dphotoperiod and 35–40 mol m^-2s^–1 photon flux density. The shoots were multiplied by (i) subculture of nodal shoot segments onto MS +0.1 mgl^-–1 IAA+1.0mgl^–1 BAPH+additives, and (ii) repeated transfer of original explant onto MS+ 0.1mgl^–1 IAA+mg l^–1 BAP+additives, at intervals of 3 weeks. Sixty to 70% of the shoots rooted when pulse treated with 100 mg l^–1 IBA in half strength MS liquid medium for 4h, and then transferred onto hormone-free half-strength agar-gelled MS basal saltmedium. Incubation in dark at 33 ± 2°C for 6d favoured root induction. In vitro hardened plants were transferred to pots.Abbreviations IAA Indole-3-aceticacid - IBA Indole-3-butyric acid - NAA -naphthaleneacetic acid - BAP 6-benzylaminopurine - Kn 6-furfurylaminopurine - 2-ip Isopentenyl adenine - B₅ Gamborg et al. (1968) medium - MS Murashige and Skoog's (1962) medium - WP Woody plant medium (Lloyd and McCown 1981)     

Agrobacterium-mediated transformation of ginseng (Panax ginseng) and mitotic stability of the inserted β-glucuronidase gene in regenerants from isolated protoplasts

Cotyledonary expiants of ginseng zygotic embryos were cocultured with Agrobacterium tumefadens strain LBA4404 harboring the binary vector pBI121 for 48 h and transferred onto MS medium supplemented with 1 mgl^–1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.1 mgl^–1 kinetin, and 100 mgl^–1 kanamycin. After 8 weeks of culture, kanamycin-resistant calli formed on the cut surfaces of cotyledonary expiants and subsequently they gave rise to numerous somatic embryos. Eight weeks after transfer onto medium containing 1 mgl^–1 each of 6-benzyladenine (BA) and gibberellic acid, most of them developed into plantlets. Southern analysis confirmed that the -glucuronidase (GUS) gene was incorporated into the genomic DNA of regenerants. Protoplasts were enzymatically isolated from transformed somatic embryo segments and cultured in liquid medium containing 60 gl^–1 myo-inositol, 1 mgl^–1 2,4-D, 0.5 mgl^–1 BA, and 0.5 mgl^–1 kinetin. Plants were regenerated from protoplasts via somatic embryogenesis. The polymerase chain reaction method revealed that 92% of the regenerants retained the GUS gene. When treated with X-glucuronide, 78% of the regenerants showed a GUS-positive response. The overall results indicate that the transgene is stably transmitted during somatic ontogeny and stably expressed in most the regenerants, whereas it may be deleted or impaired in some portion of them.Abbreviations BA 6-benzyladenine; 2,4-D,2,4-dichlorophenoxyacetic acid - DIG digoxigenine - GA₃ gibberellic acid - X-gluc X-glucuronide - GUS -glucuronidase - MS Murashige and Skoog (1962)     

Biomass and icosapentaenoic acid productivities from an outdoor batch culture of Phaeodactylum tricornutum UTEX 640 in an airlift tubular photobioreactor

An outdoor tubular photobioreactor, based on an external-loop airlift, has been used to cultivate the diatom Phaeodactylum tricornutum UTEX 640, rich in icosapentaenoic acid (EPA). The system was operated in batch mode. The specific growth rate in the exponential phase, averaged over the whole day, was 0.254 day^–1 · A 25 gl^–1 maximum biomass concentration was obtained by the end of the culture period. The dissolved oxygen generated never rose over 210% of air saturation nor was there impediment to biomass generation or inhibition of photosynthesis. Furthermore, around 2 × 10¹⁷ quanta cm^–2 s^–1 light intensity seemed to saturate the photosynthetic activity. Biomass losses during the night could be reduced to less than 5% by maintaining the culture at 16°C at night. The maximum concentration and productivity of EPA were 423 mgl^–1 and 13 mgl^–1 day^–1, respectively, at the end of the culture, although a 21-mgl^–1 day^–1 net EPA generation rate was obtained at the linear growth phase.     

Somatic embryogenesis from leaf- and petiole-derived callus of Vitis rupestris

Somatic embryogenesis from leaf- and petiole-derived calli of Vitis rupestris was obtained with an efficiency of 3.2% and 4.2% of plated explants, respectively on two combinations of 6-benzyladenine and 2,4-dichlorophenoxyacetic acid (1/0.1 and 1/1 mgl^–1) added to MS medium. Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl^–1 indole-3-acetic acid or 0.1 mgl^–1 indole-3-butyric acid added to MS or NN media. Within a 4-month culture, embryo germination occurred at a frequency of 13% of explanted embryos when chilling at 4°C was provided for two weeks and a combination of 6-benzyladenine (1 mgl^–1) with indole-3-butyric acid (0.1 mgl^–1) was added to NN medium supplemented with casein hydrolysate (250 mgl^–1). A higher frequency (51%) was obtained in a longer culture time (9 months) when only indole-3-butyric acid was present in the medium and in absence of chilling.Abbreviations BA 6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) - NN Nitsch and Nitsch (1969) - NOA 2-naphthoxyacetic acid     

Trace Metal Inventories and Lead Isotopic Composition Chronicle a Forest Fire’s Remobilization of Industrial Contaminants Deposited in the Angeles National Forest

The amounts of labile trace metals: [Co] (3 to 11 µg g⁻¹), [Cu] (15 to 69 µg g⁻¹), [Ni] (6 to 15 µg g⁻¹), [Pb] (7 to 42 µg g⁻¹), and [Zn] (65 to 500 µg g⁻¹) in ash collected from the 2012 Williams Fire in Los Angeles, California attest to the role of fires in remobilizing industrial metals deposited in forests. These remobilized trace metals may be dispersed by winds, increasing human exposures, and they may be deposited in water bodies, increasing exposures in aquatic ecosystems. Correlations between the concentrations of these trace metals, normalized to Fe, in ash from the fire suggest that Co, Cu, and Ni in most of those samples were predominantly from natural sources, whereas Pb and Zn were enriched in some ash samples. The predominantly anthropogenic source of excess Pb in the ash was further demonstrated by its isotopic ratios (²⁰⁸Pb/²⁰⁷Pb: ²⁰⁶Pb/²⁰⁷Pb) that fell between those of natural Pb and leaded gasoline sold in California during the previous century. These analyses substantiate current human and environmental health concerns with the pyrogenic remobilization of toxic metals, which are compounded by projections of increases in the intensity and frequency of wildfires associated with climate change.     

Induction of mitosis in polytene nuclei and hormonal effect on nuclear changes during callus initiation in diploid Urginea indica Kunth. (liliaceae)

Bulb scale and inflorescence explants of Urginea indica Kunth. (2n=20) were cultured in vitro on modified Murashige & Skoog's medium with different hormonal composition. Media containing 2,4-dichlorophenoxy-acetic acid (2,4-D) (2 and 4 mgl^–1) and -naphthalene-acetic acid (NAA) (2 mgl^–1) could induce callus in inflorescence explants. Combination of 2,4-D (4 mgl^–1) + NAA (2 mgl^–1) + Kinetin (2 mgl^–1) only could induce callus formation in scale explants. The bulb scale explants contained mostly diploid cells while the inflorescence explants contained cells with nuclear DNA content ranging from 2C to 64C. The lowest karyological heterogeneity was recorded in callus derived from bulb scale and in callus derived from inflorescence induced with NAA. The highest variability was recorded on media with 2,4-D alone. Induction of division, probably of the pre-existing polytenic nuclei in the inflorescence explant, has been suggested to be the cause of origin of polyploid cells in such cases.     

High frequency plant regeneration via somatic embryogenesis in cell suspension cultures of coriander (Coriandrum sativum L.)

Hypocotyl segments and zygotic embryos of coriander formed embryogenic calli at frequencies of up to 75% when cultured on MS medium supplemented with 1 mgl^–1 2,4-D. Calli were transferred to MS liquid medium with 1 mgl^–1 2,4-D to initiate cell suspension cultures. Embryogenic cells became finely dispersible in the medium as the subculture proceeded. Cultures were transferred to a nitrogen compound enriched liquid MS medium containing 2% sucrose and 0.1 mgl^–1 2,4-D, and cultured two weeks before plating on MS basal medium. Approximately 75% of cell aggregates (1 to two mm in diameter) underwent development into globular to cotyledonary somatic embryos after two weeks of plating. Most of the embryos were subsequently regenerated into plantlets. Regenerants were successfully transplanted to potting soil and grown to maturity in a phytotron.Abbreviations MS Murashige and Skoog - MS1D MS medium + 1 mgl^–1 2,4-D     

Behavioral responses of red hake,Urophycis chuss,to decreasing concentrations of dissolved oxygen

Synopsis Laboratory experiments were conducted to examine changes in behavior of red hake,Urophycis chuss, under decreasing concentrations of dissolved oxygen (DO). Since the ecological requirements of this species change with age, responses were measured for three different groups: (1) age 0+, = 89 mm total length (TL); (2) age 1+, = 238 mm TL; and (3) age 2–3+, = 397 mm TL. As DO decreased from 8–10 mg l^-1 to < 0.5 mg l^-1, changes were evident in active time, water column activity, range of horizontal movement, food searching, and agonistic behavior. Age 0+ fish were most sensitive, moving up into the water column and swimming continuously as DO levels fell below 4.2 mg l^-1. Age 2–3+ fish were the least responsive, remaining on the substrate and increasing only their range of movement at concentrations below 3 mg l^-1. Responses of age 1 + fish were variable, possibly reflecting a transition stage between the younger and older fish. Common to all groups was the decrease and eventual cessation of food searching.     

Seasonal variability of dissolved organic carbon ina Mediterranean stream

The seasonal variability of dissolved organic carbon(DOC) flux in a Mediterranean stream subjected todischarges of wide range of intensities and variabledry period was studied as a function of the hydrologicconditions, and the relationship between surface andsubsurface (hyporheic and groundwater) DOCconcentration. DOC concentration in stream water(2.6 mg l^–1 ±1.5 SD) was higher thangroundwater (1.3 mg l^–1 ± 1.2 SD) and lower thanhyporheic water (3.8 mg l^–1 ±1.7 SD),suggesting that, at baseflow, stream DOC concentrationincreases when groundwater discharges through thehyporheic zone. Storms contributed to 39% of annualwater export and to 52% of the total annual DOCexport (220 kg km^–2). A positive relationship wasobserved between Discharge (Q) and stream DOCconcentration. Discharge explained only 40% of theannual variance in stream DOC, but explained up to93% of the variance within floods. The rate of streamDOC changes with discharge change during storms (dDOC/dQ), ranged between 0 and 0.0045 C mgl^–1 s l^–1, with minimum values during Springand Summer, and maxima values in Fall and Winter.These dynamics suggest that storm inputs ofterrigenous DOC vary between seasons. During floods inthe dormant season, DOC recession curves were alwayssteeper than discharge decline, suggesting shortflushing of DOC from the leaching of fresh detritusstored in the riparian zone.     

Spatial variation in basic chemistry of streams draining a volcanic landscape on Costa Rica's Caribbean slope

Spatial variability in selected chemical, physical and biological parameters was examined in waters draining relatively pristine tropical forests spanning elevations from 35 to 2600 meters above sea level in a volcanic landscape on Costa Rica's Caribbean slope. Waters were sampled within three different vegetative life zones and two transition zones. Water temperatures ranged from 24–25 °C in streams draining lower elevations (35–250 m) in tropical wet forest, to 10 °C in a crater lake at 2600 m in montane forest. Ambient phosphorus levels (60–300 µg SRP L^–1; 66–405 µg TP L^–1) were high at sites within six pristine drainages at elevations between 35–350 m, while other undisturbed streams within and above this range in elevation were low (typically <30.0 µg SRP L^–1). High ambient phosphorus levels within a given stream were not diagnostic of riparian swamp forest. Phosphorus levels (but not nitrate) were highly correlated with conductivity, Cl, Na, Ca, Mg and SO₄. Results indicate two major stream types: 1) phosphorus-poor streams characterized by low levels of dissolved solids reflecting local weathering processes; and 2) phosphorus-rich streams characterized by relatively high Cl, SO₄, Na, Mg, Ca and other dissolved solids, reflecting dissolution of basaltic rock at distant sources and/or input of volcanic brines. Phosphorus-poor streams were located within the entire elevation range, while phosphorus-rich streams were predominately located at the terminus of Pleistocene lava flows at low elevations. Results indicate that deep groundwater inputs, rich in phosphorus and other dissolved solids, surface from basaltic aquifers at breaks in landform along faults and/or where the foothills of the central mountain range merge with the coastal plain.     

Simultaneous nitrification and denitrification in a mixed methanotrophic culture

Simultaneous nitrification and denitrification using a mixed methanotrophic culture was investigated. When both NO₃ ^–-N (108 mg l^–1) and NH₃-N (59 mg l^–1) were added into batch reactors, nitrate removal was complete within 10 h at the rate of 47 mg NO₃ ^–-N g VSS^–1 day^–1 when dissolved oxygen (DO) concentration was maintained at 2 mg DO l^–1. Ammonia removal started simultaneously with nitrate removal at a slower rate of 14 NH₃-N g VSS^–1 day^–1. No significant accumulation of nitrite or nitrate during ammonia utilization suggested the occurrence of simultaneous nitrification and denitrification.     

Impact of cefotaxime on somatic embryogenesis and shoot regeneration in sugarcane

A cephalosporin antibiotic, cefotaxime (Omnatax™) promoted somatic embryogenesis and subsequent shoot regeneration in vitro from spindle in sugarcane irrespective of the genotypes as (CoJ 83, CoJ 88 and CoJ 64) culturered on MS medium with 2,4-D (2.5 mgl⁻¹) and kinetin (0.5 mgl⁻¹). Seven different concentrations of cefotaxime (100, 200, 300, 400, 500, 600 and 700 mgl⁻¹) were tested to find the optimal concentration of cefotaxime for somatic embryogenesis from callus cultures. Among the three varieties, calli of variety CoJ 83 incubated on MS medium with 2,4-D (2.5 mgl⁻¹) + kinetin (0.5 mgl⁻¹) + cefotaxime (500 mgl⁻¹) exhibited maximum somatic embryogenesis. To improve shoot regeneration, the callus was transferred to MS medium with BAP (0.5 mgl⁻¹) + kinetin (0.5 mgl⁻¹) in combination with different levels of cefotaxime. Highest frequency of shoot regeneration was observed in callus of CoJ 83 in the presence of 500 mgl⁻¹ cefotaxime. The plantlets could be successfully hardened in polybags and transferred to soil, where they exhibited normal growth. Our results convincingly demonstrated that cefotaxime improves somatic embryogenesis from spindle and regeneration from embryogenic calli of sugarcane and hence can be strongly recommended for rapid and large scale multiplication of sugarcane.Key words: Saccharum officinarum L., leaf segments, callus, plant regeneration, antibiotic     

Overview of trace metal contamination in the Scheldt estuary and effect of regulatory measures

Seasonally, dissolved and particulate metal concentrations in the Scheldt estuary were assessed over a period of 4 years (1995–1998). High quality data were obtained following stringent analytical protocols for each step: sampling, sample treatment, sample storage and analysis. Of the 5 trace metals, Ni showed the most conservative behaviour, while Cd and Cu were clearly transferred from the particulate to the dissolved phase in the middle estuary. A substantial part of the particulate metals entering the estuarine system are lost through sedimentation. General seasonal patterns are the following: lower concentrations in spring and higher ones in winter (sometimes late fall/early winter) for dissolved metals, while in summer a pronounced rise of the longitudinal concentration profile is observed for the particulate metals. A comparison of the trace metal concentrations (dissolved and particulate) at the mouth of the estuary in 1995–1998 with those from 1981 to 1983, reveal reductions between 30 and 58%. Reductions based on direct emission measurements for almost the same period suggest reductions (dissolved + particulate) between 42 and 64%. Biomagnification (BMF) is the accumulation of a compound through the food-chain. It is in our case expressed as the ratio of the metal concentration in the organism (g g^–1, d.w.)/the metal concentration in total suspended matter (g g^–1, d.w.). Almost all BMF-values of Periwinkle, Nereis diversicolor and Macoma balthica (3 bottom organisms in the Scheldt estuary) are negative meaning that these organisms contain less heavy metals than the particulate suspended matter. For all organisms log BMFs for Pb, respectively Ni, are around –1.8, respectively –0.7. For Cd, Periwinkle shows slight enrichment (0.05) and for Cu even more (0.45), while negative values were observed for Nereis diversicolour and Macoma balthica. The latter organisms are more enriched in Zn (–0.09) than Periwinkle (–0.43).     

Physical and chemical post-dam alterations in the Jamari River, a hydroelectric-developed river of the Brazilian Amazon

The regulation of the Jamari River advanced peak floods by 1–2 months and increased dry-season discharges from 60 to 200 m³ s^–1, resulting in water levels approximately 1 m above those recorded before regulation. Daily variation in water level associated with fluctuations in electricity production by the dam propagated to the lower reaches of the tributary Candeias River. Dissolved oxygen (DO), temperature, pH, and conductivity measured over 1.5 years on three locations along the regulated and two free-flowing rivers indicated important alterations in the case of oxygen concentrations. DO levels shifted from saturated (7–8 mg l^–1) during the flood season (when the spillways were open releasing epilimnetic water) to hypoxic (1.5–3 mg l^–1) during the dry season (when the floodgates were closed and only hypolimnetic water used to run the turbines was feeding the river). Fluctuations in water level and variation in dissolved oxygen tended to be greater at the site closest to the dam, gradually attenuating downstream. Mitigation of the downstream effects of river regulation would require modifications in the operation of the dam.     

Effect of some vertebrate and invertebrate hormones on the population growth, mictic female production, and body size of the marine rotifer Brachionus plicatilis Müller

Eight vertebrate and invertebrate hormones were screened for theireffect on population growth, mictic female production, and body size of themarine rotifer Brachionus plicatilis. Growth hormone (GH) or human chorionicgonadotropin (HCG) at 0.0025–25 I.U. ml^–1 andestradiol-17 (E₂), triiodothyronine (T_{_3}),20-hydroxyecdysone (20-HE), 5-hydroxytryptamine (5-HT), gamma-aminobutyricacid (GABA) or juvenile hormone (JH) at 0.05–50 mg l^–1were added to 5-ml of Nannochloropsis oculata suspension (7×10⁶ cells ml^–1). From an initial densityof 1 individual ml^–1, rotifers were cultured with hormones for48 hours in 22 ppt seawater at 25 °C, in darkness.Rotifers were counted and classified into female types and transferred to anew algal food suspension without hormone every other day until day 8 whenbody size was measured. Population growth was significantly higher intreatments exposed to GABA (50 mg l^–1), GH (0.0025 and 0.025I.U. ml^–1), HCG (0.25 and 2.5 I.U. ml^–1), and5-HT (5 mg l^–1). E₂ caused a decrease inpopulation growth, whereas JH, 20HE, and T₃ had no effect.Mictic female production was significantly higher at 0.05 and 0.5 mgl^–1 JH and 0.05 and 5 mg l^–1 5HT. GH (0.0025 and0.025 I.U. ml^–1), E₂ (50 mg l^–1),GABA (0.5, 5 and 50 mg l^–1), and 20-HE (0.05 mgl^–1) treatments had significantly higher mictic femaleproduction only on day 8, 6, 4, and 6, respectively. T₃ andhCG had no effect on mictic female production. Lorica length increased by9.6% and 4.4% in rotifers treated with JH (0.05 mgl^–1) and GABA (5 mg l^–1), respectively. Correspondingly, lorica width increased by 8.9% and 2.6% inthese treatments. In comparison, 20-HE-, T₃-, and HCG-treatedrotifers were smaller (3.9–8.2%) and GH, 5-HT andE₂ had no effect on rotifer body size.     

Effect of pH,aeration and sucrose feeding on the invertase activity of intactS. cerevisiae cells grown in sugarcane blackstrap molasses

S. cerevisiae was grown in a blackstrap molasses containing medium in batch and fed-batch cultures. The following parameters were varied: pH (from 4.0 to 6.5), dissolved oxygen (DO) (from 0 to 5.0 mg O₂L^–1) and sucrose feeding rate. When glucose concentration (S) was higher than 0.5 g L^–1 a reduction in the specific invertase activity of intact cells (v) and an oscillatory behavior of v values during fermentation were observed. Both the invertase reduction and the oscillatory behavior of v values could be related to the glucose inhibitory effect on invertase biosynthesis. The best culture conditions for attainingS. cerevisiae cells suitable for invertase production were: temperature=30°C; pH=5.0; DO=3.3 mg O₂L^–1; (S)=0.5 g L^–1 and sucrose added into the fermenter according to the equations: (V–V_o)=t²/16 or (V–V_o)=(V_f–V_o)·(e^0.6t–1)/10.This work was supported by FAPESP     

Shoot regeneration of hybrid seed geranium (Pelargonium x hortorum) and regal geranium (Pelargonium x domesticum) from primary callus cultures

Procedures have been developed that increase the rate of shoot regeneration of hybrid seed geranium from month-old primary callus cultures. Hybrid geranium callus tissue covered with green nodular structures was initiated by placing shoot tip explants on solidified Murashige & Skoog medium (MS) supplemented with 2.0 mgl^-1 zeatin and 1.9 mgl^-1 indoleacetic acid. Hybrids Red Orbit, White Orbit and Scarlet Orbit were shown to produce 5–50 shoot primordia per explant when callus was initiated on this medium. Regal geranium callus was initiated by placing leaf explants on MS medium supplemented with 2.0 mgl^-1 6-benzylaminopurine and 2.0 mgl^-1 naphthaleneacetic acid. Regal geranium cultivars Tiny Tot and Lavender Grand Slam were shown to produce between 2–50 shoot primordia per explant when initiated on the same medium.