Double-phase in vitro culture using sorbitol increases shoot proliferation and reduces hyperhydricity in Japanese pear

This study aimed to improve in vitro shoot proliferation efficiency without inducing hyperhydricity in Japanese pear. The shoot number increased at 2.5–10.0 mg l⁻¹ benzyladenine (BA), while shoot fresh mass increased at 1.0 and 2.5 mg l⁻¹ BA. Different macroelement formulation did not affect shoot proliferation, but adding activated charcoal (AC) to the medium inhibited markedly the production of axillary shoots and biomass and many shoots were hyperhydric. Different carbon sources (CS) significantly increased the shoot number and fresh mass, with the best results for shoot proliferation at 20–30 g l⁻¹ sorbitol. With gelling agents, the shoot number increased at 0.4 and 0.6% agar and 0.3% gellan gum, while fresh mass increased at 0.4% agar. The hyperhydric explants were more than 30% at 0.4–0.6% agar and at any concentration of gellan gum. The improved culture (woody plant (WP) supplemented with 20 g l⁻¹ sorbitol, 0.1 mg l⁻¹ 3-indolyl-butyric acid (IBA), 2.5 mg l⁻¹ BA and 0.8% agar) and double-phase culture (the same medium using a double-phase liquid-gelling agent solidified culture system) produced a higher number of axillary shoots than the conventional culture (1/2MS supplemented with 0.1 mg l⁻¹ IBA, 1.0 mg l⁻¹ BA, 30 g l⁻¹ sucrose and 0.8% agar), moreover, double-phase culture had a higher fresh mass than the other cultures.     

Prohexadione-calcium affects growth and flowering of petunia and impatiens grown under photoselective films

The response of petunia (Petunia x hybrida Vilm.-Andr. ‘Countdown Burgundy’) and impatiens (Impatiens wallerana Hook ‘Accent Orange Tempo’) to Prohexadione-calcium was evaluated under a clear and a far-red light absorbing greenhouse (A_FR) film to investigate the dosage effect of Prohexadione-Ca and to determine if it can overcome the flowering delay under FR deficient greenhouse environments. Prohexadione-Ca reduced stem elongation of petunia and impatiens under A_FR and clear films when applied 3 weeks after germination. Late applications were less effective. In both crops, main stem length decreased in a quadratic pattern as the concentration of Prohexadione-Ca increased. Under both films, 50–100 mg l⁻¹ Prohexadione-Ca resulted in ≈30% shorter petunia plants. Greater concentrations (500 and 1000 mg l⁻¹) resulted in excessively short plants (over 70%). Prohexadione-Ca at 100 mg l⁻¹ delayed anthesis of petunia by 8 and 3 days under the clear film and the A_FR film, respectively during less inductive photoperiods but had no effect during inductive photoperiods. In impatiens, Prohexadione-Ca at 100 mg l⁻¹ delayed anthesis over 10 days under clear or A_FR film. Greater concentrations (200 and 300 mg l⁻¹) inhibited flowering of impatiens. Prohexadione-Ca treatments significantly affected flower color development. Untreated petunia plants had dark burgundy flowers. Prohexadione-Ca treatment increased L*, a*, and C* values and decreased hue angle indicating that the flowers were faded. Flowers of untreated impatiens plants were bright orange color. Prohexadione-Ca at 100 mg l⁻¹ increased L* value and decreased a*, b*, and C* values indicating that significant petal fading had occurred. Flowers of treated plants were nearly white under both films. Although effective in height control, loss of color would be a major limitation to the use of Prohexadione-Ca on flowering crops.     

Factors affecting the shoot multiplication of Persian walnut (Juglans regia L.)

A number of experiments were conducted to identify suitable procedure for in vitro shoot multiplication of Persian walnut (Juglans regia L.). Three different nutrient media (DKW, MS and WPM) and three different gelling agents (Phytagel, Difco Bacto agar and a mixture of Phytagel and Difco Bacto agar) were studied in the first experiment. Driver and Kuniyuki walnut (DKW) medium solidified with 2.2 g l⁻¹ Phytagel was found optimum. Performance of explants was better on DKW medium than on MS and WPM. The DKW and MS media were not significantly different from each other, but both of them were significantly better than WPM, which was a very poor medium for this species. Phytagel alone was significantly better than Difco Bacto agar or Phytagel combined with Difco Bacto agar. In another experiment different concentrations of BA were studied. Medium containing 1.0 mg l⁻¹ BA and 0.01 mg l⁻¹ IBA was the best, although medium containing 0.6 and 0.8 mg l⁻¹ BA were also successful, and utilisation of 0.4 mg l⁻¹ BA and 0.01 mg l⁻¹ IBA was optimum for shoot elongation. Application of different kinds of auxins (IAA, IBA and NAA at 0.01 or 0.1 mg l⁻¹) with 1.0 mg l⁻¹ BA were also studied. Media containing IBA were significantly better than media containing IAA for shoot fresh weight, but neither of them was significantly different from media containing NAA. Application of 0.01 mg l⁻¹ or 0.1 mg l⁻¹ auxin, with 1.0 mg l⁻¹ BA, was not significantly different for shoot multiplication of Persian walnut. The morphology of shoots on media containing 0.01 mg l⁻¹ IBA was the best.     

An investigation on the effect of different plant growth regulating compounds in in vitro shoot tip and node culture of lemon seedlings

The possible application of some less commonly used in vitro growth regulating compounds is outlined. A number of treatments were applied to determine the best way of inducing in vitro shoot proliferation and rooting on a modified Driver–Kuniyuki [HortScience 19 (1984) 507] basal medium of lemon (Citrus limon (L.) Burm, f. cv. Interdonato) seedlings. 6-Benzyladenine (BA) alone (1, 2 and 4 mg l⁻¹) and in combination with either orange juice (10%, v/v), silver nitrate (3 mg l⁻¹), gibberellic acid (GA₃) (0.1 mg l⁻¹ at the establishment stage and 0.5 mg l⁻¹ at all combinations during the proliferation stage) or abscisic acid (ABA) (0.2 mg l⁻¹ only at the establishment stage) were used to stimulate shoot formation during the establishment and the proliferation stage. The combination of BA with ABA gave a high rate of shoot formation, while GA₃ and silver nitrate enhanced shoot elongation. When these shoots were transferred to the rooting stage, the effect of application of two different auxins (indole-3-butyric acid (IBA) and α-napthaleneacetic acid) was examined, as well as different methods of application (auxin added to the basal medium and auxin application by dipping the base of the explant in auxin solution). Dipping the base of the explants in a 50% ethanol solution of IBA at 1000 mg l⁻¹ for 5 s resulted in 80% rooting with subsequent 90% survival of these explants, during acclimatization under mist.     

Effect of phytohormones on in vitro shoot multiplication and rooting of lime Citrus aurantifolia (Christm.) Swing

Multiple shoots were produced from node explants of lime (Citrus aurantifolia (Christm.) Swing) on MS medium supplemented with 6-benzylaminopurine (BAP), 6-furfurylaminopurine (kinetin) and α-naphthaleneacetic acid (NAA). The highest number of shoots, nine shoots per node, were produced on a medium containing 2 mg l⁻¹ BAP (8.8 μM), 1 mg l⁻¹ kinetin (4.6 μM) and 1 mg l⁻¹ NAA (5.4 μM). Depending on the concentration of BAP and kinetin, NAA either inhibited, stimulated or did not affect shoot multiplication, which also depended on the cytokinin level. Maximum shoot length was obtained from treatments containing 0.5 mg l⁻¹ BAP (2.2 μM) combined with 1 mg l⁻¹ kinetin (4.6 μM) and 0.5 mg l⁻¹ NAA (2.7 μM). The largest leaves of resultant shoots were produced on a medium containing 0.5 mg l⁻¹ each of kinetin (2.3 μM) and NAA (2.7 μM). Transferring in vitro shoots to rooting media containing indole-3-butyric acid (IBA) and NAA produced complete plantlets. The highest rooting percentage was obtained on a medium containing either 1 mg l⁻¹ NAA (5.4 μM) alone or 0.5 mg l⁻¹ NAA (2.7 μM) combined with 2 mg l⁻¹ IBA (9.6 μM), whereas the highest number of roots were produced on a treatment containing both 2 mg l⁻¹ NAA (10.8 μM) and 2 mg l⁻¹ IBA (9.6 μM). Roots elongated most on treatments containing 0.5 mg l⁻¹ of either NAA (2.7 μM) or IBA (2.4 μM). Shoot growth associated with the rooting phase was the highest in response to 2 mg l⁻¹ IBA (9.6 μM) or 0.5 mg l⁻¹ NAA (2.7 μM). Plantlets that survived acclimatization, 82%, exhibited normal growth in soil under greenhouse conditions.     

Establishment and plant regeneration of somatic embryogenic cell suspension cultures of the Zingiber officinale Rosc.

Somatic embryogenic cell suspension cultures of four ginger cultivars were established. Somatic embryogenic calli were induced from ginger shoot tips on MS agar medium supplemented with 1.0 mg l⁻¹ 2,4-D and 0.2 mg l⁻¹ Kn, which contained only half concentration of NH₄NO₃. Rapid-growing and well-dispersed suspension cultures were established by subculturing this kind of callus in the same liquid MSN medium. The suspension cultures (about 1–2 mm in diameter) were placed on the MSN agar medium for callus proliferation. Thereafter embryogenic callus (1.5 cm²) was transferred to solid media (MS + 0.2 mg l⁻¹ 2,4-D + 5.0 mg l⁻¹ BA + 3% sucrose + 0.7% agar). Somatic embryos produced shoots and roots, and shoots developed into complete plantlets on solid MS medium supplemented with 3.0 mg l⁻¹ BA and 0.1 mg l⁻¹ NAA. The relationship between the DW of suspension cultures and pH changes in medium is also discussed. The suspension cultures still kept their vitalities after subculture for 8 months.     

The influence of NaCl salinity on growth,yield and fruit quality of strawberry cvs. ‘Elsanta’ and ‘Korona’

In a 2-year field study, strawberry cvs. ‘Elsanta’ and ‘Korona’ were exposed to three different levels of NaCl salinity supplied as aqueous solutions characterised by electrical conductivities of 0.3 dS/m, 2.6 dS/m, and 5.1 dS/m. Salinity in the rhizosphere reduced plant growth by up to 44% in ‘Korona’ and 90% in ‘Elsanta’. A rather distinct cultivar difference represented the reduction in leaf area per plant of 85% in the second year of experiment in ‘Elsanta’ compared to 29% in ‘Korona’. Strawberry can be regarded as a Na⁺ excluder, because Na⁺ content of both strawberry cultivars remained below 3 mg g⁻¹ dry mass at all salinity levels. Cl⁻ content increased considerably, up to 70 mg g⁻¹ dry mass in ‘Korona’ and 80 mg g⁻¹ dry mass in ‘Elsanta’ plants. ‘Korona’ retained most of its Cl⁻ in roots and crowns, whereas in ‘Elsanta’ the maximum was detected in petioles. ‘Korona’ was able to accumulate up to 33% higher Cl⁻ content in the roots than ‘Elsanta’. Macronutrient deficiency due to NaCl salinity was not observed and in comparison to ‘Elsanta’, higher Cl⁻ content in roots of ‘Korona’ did not coincide with an impairment of macronutrient uptake. Salinity stress reduced fruit yield by up to 27% in ‘Korona’ and 64% in ‘Elsanta’. Fruit quality, characterised as taste, aroma, and texture by a consumer-type panel, decreased by more than 24% in ‘Elsanta’, but in ‘Korona’ differences were insignificant. Total soluble solids (Brix) and the ratio Brix/TA (TA, titratable acid) decreased significantly by about 20% in ‘Korona’ and 35% in ‘Elsanta’. To summarise, the ability of ‘Korona’ to retain Cl⁻ in the root system more effectively than ‘Elsanta’ resulted not only in a 41% lower leaf Cl⁻ content at the highest salinity level and a better growth under NaCl stress, but also in a relatively higher fruit yield and fruit quality.     

Foliar treatment of Mn deficient ‘Washington navel’ orange trees with two Mn sources

The objective of this study was the comparison of the effect of two Mn sources (MnSO₄·H₂O, MnEDTA) which were applied at various concentrations (0, 200, 400, 800, and 1200 mg Mn l⁻¹) to the leaves of ‘Washington navel’ orange trees in order to correct Mn deficiency.One hundred and seventy days after the foliar application of Mn solutions, the mean Mn concentrations in the leaves treated with MnSO₄·H₂O (200, 400, 800 or 1200 mg Mn l⁻¹) or MnEDTA (400, 800 or 1200 mg Mn l⁻¹) were significantly higher than those of the control leaves. Manganese sulfate (MnSO₄·H₂O) was more effective than MnEDTA regarding the improvement of the leaf Mn concentrations of the trees, when applied at equal Mn concentrations. Finally, the leaf Mn concentrations were in the sufficiency range (>25 mg kg⁻¹ d.w.), only after the application of 800 or 1200 mg Mn l⁻¹ as MnSO₄·H₂O.     

Arsenic as a factor affecting virus infection in tomato plants: changes in plant growth,peroxidase activity and chloroplast pigments

The influence of arsenic and Cucumber mosaic virus (CMV), applied separately and simultaneously on young tomato plants was studied. The plants were cultivated in containers under glasshouse conditions. Four main variants were arranged. The first one was without additional As pollution of soil, named as a control, and the other three variants, with As added at 25, 50 and 100 mg kg⁻¹ to dry soil respectively. Half of the plants in each experimental container were inoculated with CMV and the rest uninoculated. A clear response in plant behavior under the conditions of biotic and abiotic stress was estimated. Both arsenic and virus infection had a negative effect on tomato plants by limiting the growth of their roots and above growth parts. The changes in roots were more significant than of stems. Virus infection was a stronger stress factor than arsenic applied at levels of 25 and 50 mg kg⁻¹. The effect of each stress factor applied separately was enhanced in cases of their simultaneous application. The strongest negative effect was manifested in the infected plants, treated with excess arsenic of 100 mg kg⁻¹. It was established that the infection, caused by CMV in tomatoes, was affected by the presence of arsenic in the soil and concentration of the latter. Doses of 25 and 50 mg kg⁻¹ were favorable for infection development, while the dose of 100 mg kg⁻¹ was an inhibitor.Virus infection induced stronger specific peroxidase activity (SPOA) than As treatment. The combination of both stress factors reduced the positive peroxidase response caused by virus infection. Arsenic at rate 50 and 100 mg kg⁻¹, virus infection and the combination of both stress factors at 25 mg kg⁻¹ reduced chlorophyll a, chlorophyll b and carotenoid content The virus infection in cases of the higher arsenic doses reduced the As effect. There was an interaction between the two effects of biotic and abiotic stress. When arsenic and virus infection were applied simultaneously, they caused modification of the effect of each stress on the plants, when applied separately.     

In vitro induction of tetraploid ginger (Zingiber officinale Roscoe) and its pollen fertility and germinability

In vitro induction of tetraploid ginger (Zingiber officinale Roscoe) and its pollen fertility and germinability were investigated. The growth of shoot tip cultures on agar MS medium containing 2.0 mg l⁻¹ BA was greater than that of similar cultures in liquid MS medium with the same BA concentration. In liquid medium, the combinations of 0.5, 1.0, or 2.0 mg l⁻¹ BA with 0.05 mg l⁻¹ NAA tended to enhance the growth of the cultures. The efficiency of tetraploid induction was assessed by treating shoot tip explants on agar or in liquid MS medium containing 2.0 mg l⁻¹ BA, 0.05 mg l⁻¹ NAA, and 0.2% (w/v) colchicine for 4, 8, 12, and 14 days. The total number of tetraploids induced on solid medium was 18, but only five in liquid medium. On both media, the colchicine treatment for 8 days gave the maximum level of tetraploid induction. Therefore, it was found that the treatment of shoot tip explants on agar medium containing 2.0 mg l⁻¹ BA, 0.05 mg l⁻¹ NAA, and 0.2% (w/v) colchicine for 8 days was the most efficient way of inducing tetraploid ginger. Induced tetraploid strains, ‘4× Kintoki’, ‘4× Sanshu’, and ‘4× Philippine cebu 1’, had higher pollen fertility and germinability than the diploid counterparts, i.e., in the diploid strains, pollen fertility ranged from 0.3 to 6.2% and the germination rate from 0.0 to 0.1%, while in the tetraploid strains, pollen fertility ranged from 27.4 to 74.2% and the germination rate from 4.8 to 12.9%.     

Influence of nickel-contaminated soils on fenugreek (Trigonella corniculata L.) growth and mineral composition

Leafy vegetables accumulate higher amount of heavy metals like nickel (Ni) due to their more leafy vegetative growth. Therefore, a screenhouse experiment was conducted using an alkaline sandy loam soil equilibrated with graded levels of Ni (0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 and 300 mg kg⁻¹ soil) to assess the Ni accumulation pattern and its influence on growth and micronutrient distribution in fenugreek plants. Green as well as the dry matter yields of fenugreek increased slightly up to 20 g Ni kg⁻¹ soil but decreased significantly with the application ≥40 mg Ni kg⁻¹ soil. Crops showed characteristic toxicity symptoms of interveinal chlorosis in pots receiving ≥40 mg Ni kg⁻¹ soil. While the total content of Ni in the plant tissues increased consistently with increasing rates of applied Ni, the roots accumulated much higher amount of this element compared to the shoot. The content of Fe in plants showed an increase whereas that of Cu and Zn experienced a decrease with the rise in the applied Ni.     

The effect of mycorrhizal symbiosis on the development of micropropagated artichokes

In this work, microrosettes of Cynara cardunculus L. var. scolymus Fiori of the “catanese” type were subcultured in a medium supplemented with 6-benzylaminopurine (BAP) (0.05 mg l⁻¹). For root induction, indoleacetic acid (IAA), α-naphthalene acetic acid (NAA) and indole-3-butyrric acid (IBA) were used at three concentrations: 2, 5 and 10 mg l⁻¹. The highest percentage of rooted shoots was aided by the presence of 10 mg l⁻¹ IAA.Once transplanted in pots, the plantlets were inoculated with 10 g Glomus viscosum strain A6 (AM fungus). Acclimatisation was clearly facilitated by the addition of the AM fungus. Indeed, the mycorrhizal plantlets registered a survival of between 90 and 95% for the rooting shoots and 60% for the non-rooting shoots.The botanical characterization of the material produced was carried out in field and was based on several morphological and productive parameters. Data collected confirm the characteristics of the original cultivar, the efficiency of the in vitro propagation material and the possibility of using this technique in early types of artichoke.     

Micropropagation of Karonda (Carissa carandas) through shoot multiplication

Shoot tips from field grown, mature plants of Carissa carandas cv. Pant Sudarshan were cultured on Murashige and Skoog’s (MS) basal medium supplemented with benzyladenine (BA) and indolebutyric acid (IBA) during different seasons. The maximum sprouting rate was obtained with 1.5 cm long explant collected in spring season (February–March) followed by those collected in summer season (April–June). Shoot proliferation was highest on MS basal media supplemented with 3.0 mg l⁻¹ BA. Rooting of microshoots was noted to be the best in 1/2 MS plus 0.8 mg l⁻¹ IBA and 0.2 mg l⁻¹ naphthalene acetic acid (NAA). The rooted plantlets were successfully acclimatized in vermiculite:sand:soil (1:1:1) potting mixture.     

Seasonal changes in CO2 assimilation in leaves of five major Greek olive cultivars

Leaf CO₂ assimilation rate, stomatal conductance (g_s), internal CO₂ concentration (C_i), chlorophyll (a + b) content, specific leaf weight (SLW) and stomatal density were measured during the season, under field conditions, for five major Greek olive cultivars, ‘Koroneiki’, ‘Megaritiki’, ‘Konservolia’, ‘Lianolia Kerkiras’, and ‘Kalamon’, with different morphological and agronomic characteristics and diverse genetic background. Measurements were taken from current-season and 1-year-old leaves, and from fruiting and vegetative shoots, throughout the season, from March to November in years 2001 and 2002. CO₂ assimilation rates showed a substantial seasonal variation, similar in all cultivars, with higher values during spring and autumn and lower values during summer and late autumn. Stomatal conductance (g_s) followed similar trends to leaf CO₂ assimilation rates, increasing from March to July, following by a decrease during August and increasing again in autumn. ‘Koroneiki’ had the highest leaf CO₂ assimilation rate and g_s values (21 μmol m⁻² s⁻¹ and 0.45 mol m⁻² s⁻¹, respectively) while ‘Lianolia Kerkiras’ and ‘Kalamon’ showed the lowest leaf CO₂ assimilation rate and g_s values (13–14 μmol m⁻² s⁻¹ and 0.22 mol m⁻² s⁻¹, respectively). One-year-old leaves had significantly higher leaf CO₂ assimilation rate than current-season leaves from April to June, for all cultivars. From August and then, leaf CO₂ assimilation rate in current-season leaves was higher than in 1-year-old leaves. There were no significant differences in leaf CO₂ assimilation rate between fruiting and vegetative shoots. Total chlorophyll (a + b) content increased with leaf age in current-season leaves. In 1-year-old leaves chlorophyll content increased in spring, then started to decrease and increased slightly again late in the season. Chlorophyll content was higher in 1-year-old leaves than in current-season leaves throughout the season. Total specific leaf weight (SLW) increased throughout the season for all cultivars. Stomatal density in lower leaf surface was lowest for ‘Koroneiki’ (399 mm⁻²) and highest for ‘Megaritiki’ (550 mm⁻²). Our results showed differences in leaf CO₂ assimilation rate among the five different olive cultivars, with a diverse genetic background, ranging from 12 to 21 μmol m⁻² s⁻¹. From the five cultivars examined, ‘Koroneiki’, a drought resistant cultivar, performed better and was able to maintain higher leaf CO₂ assimilation rate, even under high air vapor pressure deficit. All cultivars had a pronounced seasonal variation in leaf CO₂ assimilation rate, affected by date of the year, depending on ambient conditions. The high temperatures and high air vapor pressure deficit occurring during summer caused a reduction in leaf CO₂ assimilation rate in all cultivars. Leaf CO₂ assimilation rate was also affected by leaf age for all cultivars, with old leaves having significantly higher leaf CO₂ assimilation rate than young leaves early in the season.     

Effects of IAA and BA on development of globular,heart- and torpedo-stage embryos from cell suspensions of three grape genotypes

In order to initiate cell and embryo suspensions, embryogenic calluses derived on NN solid medium with 2,4-D and BA from petioles of in vitro grown plants of three interspecific grapevine hybrids were cultured in three versions of liquid NN medium: (1) without growth regulators, (2) 0.1 mg l⁻¹ IAA and (3) 0.5 mg l⁻¹ BA. Cell and embryo suspensions were incubated two and three times in these versions of liquid media in various combinations. Incubating suspensions two times in hormone-free media and/or with 0.1 mg l⁻¹ IAA led to formation of globular embryos in the three cultivars studied and small numbers of heart-stage embryos in ‘Podarok Magaracha’ and ‘Intervitis Magaracha’. Numerous heart-stage embryos developed in ‘Intervitis Magaracha’ and ‘Podarok Magaracha’ when the suspensions had been initiated in medium with 0.5 mg l⁻¹ BA, and in ‘Bianca’ this was achieved after two incubations in the above medium. Torpedo-stage embryos were formed after subculturing heart-stage embryo suspensions in medium with 0.1 mg l⁻¹ IAA in all study cultivars. If only small numbers of embryos of a certain developmental heart- or torpedo- stage were formed, such cell and embryo suspensions need to be repeatedly subcultured in liquid medium with specific growth regulators to enable this process.     

Shoot induction and plant regeneration from receptacle tissues of Lilium longiflorum

An efficient system has been developed for the in vitro plant regeneration of Lilium longiflorum Thunb. by culturing receptacle sections from flower buds. The sections were cultured on one-half MS medium plus 30 g l⁻¹ sucrose, 8 g l⁻¹ agar, 5.4 μM NAA or 4.9 μM IBA plus 2.2 μM BAP. A section size of 3–4 mm was found to be optimal. After 60 days an average of 41 shoots were formed per explant. More vigorous shoots were obtained by subculturing on hormone-free medium with 20 g l⁻¹ sucrose. Rooting occurred on one-half MS medium with 1.1 μM NAA. Rooted plants were hardened-off in a greenhouse for two months, and normal flowering plants were produced.     

The impact of photoperiod and irradiance on flowering of several herbaceous ornamentals

Forty-one herbaceous species were grown under short-days (8 h photoperiod, ambient irradiance averaged 12–13.2 and 6.4–8.3 mol m⁻² day⁻¹ for Experiments I and II, respectively) with or without supplemental high-pressure sodium lighting (+50, 100, or 150 μmol m⁻² s⁻¹); or under long-days delivered using natural day lengths and irradiance with night interruption lighting (2200–0200 h at 2 μmol m⁻² s⁻¹ from incandescent lamps) or under ambient daylight plus supplemental irradiance during the day and as a day extension to 18 h (0800–0200 h) with supplemental high pressure sodium lighting (+50, 100, or 150 μmol m⁻² s⁻¹) to identify the impact of photoperiod and irradiance on flowering of each species. Days to first open flower, leaf number below first flower, and mean dry weight gain per day (MDWG) were measured when the first flower opened. Twenty-seven species were photoperiodic with examples of five photoperiodic response groups represented: obligate short-day (2), facultative short-day (5), obligate long-day (16), facultative long-day (4); 13 were day neutral (no photoperiod response in flowering). One species, Salvia sclarea L., did not flower. A facultative irradiance response was observed with 10 species; 28 species were irradiance indifferent; 2 had delayed flowering as irradiance increased. Photoperiod affected MDWG of 30 species. Increasing irradiance affected MDWG with 14 species. Photoperiod interacted with irradiance to affect MDWG of 11 species. Cobaea scandens had the greatest MDWG (0.40 g day⁻¹) while Amaranthus hybridus had the least MDWG (0.01 g day⁻¹) across photoperiod and irradiance levels.     

Rose productivity and physiological responses to different substrates for soil-less culture

Cultivation of roses in various soil-less media was studied with the aim to identify the optimum soil condition for rose production. Madelon roses grafted on rootstock of Rosa indica var. major were transplanted to polyethylene bags containing zeolite and perlite (at ratios of 25z:75p, 50z:50p, 75z:25p and 100z:0p, v/v) in a climate-controlled greenhouse. Net photosynthesis (A_net), stomatal conductance (g_s) and water use efficiency (WUE) of roses were followed for 5 months. Flower production and quality were recorded in three flowering flushes during a 5-month period. Analysis of variance of repeated measurements showed that even though the overall A_net did not differ among treatments (average 18.7 μmol m⁻² s⁻¹), trends in A_net seasonality for roses in 25z:75p substrate differed significantly from those in 50z:50p, 75z:25p or 100z:0p. Stomatal conductance did not show any significant seasonality or trends in response to substrate mixtures, averaging 0.89 mol m⁻² s⁻¹. Water use efficiency was significantly lower for roses in 25z:75p than in 100z:0p mixtures (1.8 ± 0.15 and 2.0 ± 0.13 μmol m⁻² s⁻¹ CO₂/mmol m⁻² s⁻¹ H₂O, respectively). Cumulative production of rose plants did not differ among substrate mixtures. Productivity significantly differed among flower stem classes. Stem class I (>70 cm) and class V (≤30 cm) exhibited the least production, contributing to only 7.6 and 3.7% of the total production, respectively. The highest productivity was observed in classes III (51–60 cm) and IV (31–50 cm), contributing to the bulk of productivity (68.4%). Class II contributed a 20.3% of the production. Results showed that zeolite and perlite acted as inert materials. Zeolite did not exert any positive effect on productivity, in contrast to what has been reported in literature recently. Use of perlite resulted in a little improvement in photosynthesis, however this improvement was not reflected by a significant increase in production.     

Methyl jasmonate and CMN-pyrazole applied alone and in combination can cause mature orange abscission

Methyl jasmonate (MJ) and CMN-pyrazole alone and in combination were applied to ‘Hamlin’ and ‘Valencia’ orange trees (Citrus sinensis (L.) Osbeck) in seven separate experiments in the 1998/1999 and 1999/2000 growing seasons to induce abscission of mature fruit for mechanical harvesting. MJ alone significantly reduced fruit detachment force (FDF) of ‘Hamlin’ oranges at concentrations of 10 mM and higher. However, MJ at 20 and 100 mM caused significant defoliation. For ‘Hamlin’ oranges, the most effective treatments were those combining 10 mM MJ and 50 mg l⁻¹ CMN-pyrazole, and 20 mM MJ and 25 mg l⁻¹ CMN-pyrazole. ‘Valencia’ oranges did not respond to 10 mM MJ treatments alone or in combination with CMN-pyrazole, and required MJ at 20 mM in combination with CMN-pyrazole to loosen this late-maturing variety to below 50 N, but excessive defoliation occurred.     

Factors controlling high efficiency adventitious bud formation and plant regeneration from in vitro leaf explants of roses (Rosa hybrida L.)

Studies were conducted to improve adventitious bud regeneration in roses (Rosa hybrida L.), specifically to extend the protocol to different genotypes and to initiate production of multiple shoots per explant. The best results were obtained by using a two-stage procedure where excised leaflets were incubated on Murashige and Skoog (MS) (1962) induction medium with 6.8 μM TDZ plus 0.49 μM IBA in the dark for 7 days and subsequently transferred to an MS-based regeneration medium with 2.22 μM BA plus 0.049 μM IBA exposed to a PPFD of 15 μmol m⁻² s⁻¹ PAR. Bud formation capacity was also significantly affected by the genotype and the environment, such as the use of bottom cooling creating a lower RH in the vessel. The addition of silver nitrate to the induction medium also significantly improved the percentage of regeneration in three genotypes tested. Regenerated shoots failed to elongate when transferred to MS proliferation medium containing 0.5 mg l⁻¹ BA, however maximum bud development and elongation were achieved when kinetin in the range 1–2 mg l⁻¹ was used. Elongated shoots were excised and rooted best on zero growth regulator half-strength MS modified medium. Rooted plantlets were acclimatized under greenhouse conditions for evaluation of somaclonal variation.