Nitrogen and calcium nutrition of Petunia × hybrida ‘Coral Sea’

The effects of N and Ca nutrition on plant growth and shoot elemental content of Petunia × hybrida Hort. Vilm. - Andr. ‘Coral Sea’ were evaluated. Nitrogen and Ca were applied separately or in combination in three experiments: (1) N at 0, 100, 200 or 400 mg l^?1; (2) Ca at 0, 75, 150 or 300 mg l^?1; (3) N at 0 or 100 mg l^?1 and Ca at 0 or 150 mg l^?1 combined factorially. Shoot and root dry weights, branch length and flower number were highest when plants received 100 mg l^?1 N. Plants treated with 150 mg l^?1 Ca had the highest shoot and root dry weights. Branch length was maximal at 300 mg l^?1 Ca.Nitrogen and Ca interacted to increase shoot dry weights, branch number and length, leaf area and flower number. Increasing N concentrations increased N and decreased P, Mn and Zn shoot contents. Calcium content of shoots increased while N, P and Mg decreased in response to increasing applications of Ca to petunia plants. Minimal N and Ca tissue concentrations for optimal P. × hybrida growth were 3.3 and 0.67%, respectively.     

Nitrogen fertilization under saline conditions in tomato and cucumber plants

Summary

Tomato (Lycopersicon esculentum, Mill.) and cucumber (Cucumis sativus, L.) seedlings were grown in a growth chamber in 4 litre containers filled with nutrient solutions. Four experiments were conducted with four NaCl levels factorially combined with four N levels. The concentrations of NaCl were 4, 25, 50, and 100 mmol l^?1 and 4, 16, 32, and 64 mmol l^?1 in the tomato and cucumber experiments respectively. The N levels in allexperiments were 2, 6, 10 and 15 mmol l^?1 added as NO₃^? or as NH₄⁺ + NO₃^? (2:1). Salinity had a statistically significant negative effect on all responses considered, although the effect was N source-dependent. Addition of N enhanced development of shoot and root dry weights of both species. The optimum N concentration in the nutrient solution varied between 6 and 10 mmol l^?1, although the most appropriate N fertilizer varied with species. Leaf Cl^? concentration decreased in both species when NO₃^? was used as the N source, whereas it increased in the comparative tissues of plants fed with NH₄⁺ + NO₃^?. The effect of treatments on the other nutrients show effects of excess of NaCl, as the form of N source interferes with the uptake of essential nutrients which may cause nutritional disorders.     

Effects of nitrogen and phosphorus nutrition on the growth of asparagus seedlings

N was applied at 50, 100 or 150 mg l^?1 in factorial combination with P at 7.5, 15 or 22.5 mg l^?1 to asparagus seedlings. There were 6 successional harvests. N and P increased shoot dry weight by increasing mean dry weight and number of shoots. Increasing P had no effect on shoot growth at 50 mg l^?1 N. N increased root dry weight (crown and roots) by increasing root number, whereas P decreased root dry weight due to a decrease in mean root dry weight. N increased total plant dry weight, but P had no effect. N and P increased the partitioning of dry weight to the shoots, while partitioning to the roots increased with time. Plant analysis revealed that 2.6–2.7% N and 0.29–0.36% P, on a dry-weight basis, were present in the shoots at the later harvests with the higher concentrations of N and P. 100–150 mg 1^?1 N in combination with 15 mg l^?1 P produced a seedling suitable for transplanting into commercial fields at 6 weeks from emergence.     

Meristem culture and micropropagation of a variety of ginger (Zingiber officinale Rosc.) with a high yield of oleoresin

Summary

Meristems of ginger with or without leaf primordia were induced to form shoots on three-quarter strength Murashige-Skoog’s (MS) medium containing sucrose 6%, coconut milk (CM) 20%, ascorbic acid (AA) 100 mg l^?1, glutamine (GL) 400 mg l^?1, activated charcoal (AC) 250 mg l^?1, 6-benzylaminopurine (BAP) 0.5 mg l^?1, indolebutyric acid (IBA) 0.4 mg l^?1 and agar 0.8%. Meristem-derived shoots exhibited consistent multiplication on three-quarter strength MS medium containing sucrose (3%), AA (100 mg l^?1), AC (100 mg l^?1), BAP (4–5 mg l^?1) and agar (0.8%). Liquid media (agitated or static) were less effective than a solid (agar-gelled) medium for micropropagation. Kinetin and naphthalene acetic acid (NAA) incorporated at various levels (0.01–0.8 mg l^?1) with or without added BAP and IBA neither improved plantlet formation nor enhanced shoot multiplication. The in vitro plants were successfully established in vivo and the rhizome yield was comparable with that of plants grown by conventional methods.     

The effect of nitrogen,volume of media,plant density and module shape on the growth of asparagus seedlings

There were three experiments in which the seedlings were raised in media contained in plastic cells. In Experiment 1, 150 or 200 mg l^?1 N were applied in factorial combination with cell volumes of 20, 32 or 50 cm³, There were 6 successional harvests. All cells were 7.5 cm deep and cylindrical in shape. Experiment 2 compared these 3 cell volumes at densities of 983, 1532 or 2440 plants per m². Cell diameters restricted the 50-cm³ cell to the 983, and the 32-cm³ cell to the 983 and 1532 plants per m² densities. 200 mg l^?1 N increased shoot growth, but not root growth. This increase in shoot growth was not considered advantageous. Seedling growth increased with increases in cell volume, but there were no responses to changes in plant density. Experiment 3 compared a cell having an inverted pyramid shape with a cylinder. The cylinder, despite holding 20% less media, produced the larger seedlings.     

Regeneration of Plants from Alginate-encapsulated Shoot Tips of Myrtle (Myrtus communis L.)

Myrtle growing naturally in the Mediterranean Region in Turkey, is among the economically important plant species. The present study emphasized on in vitro conservation of M. communis by encapsulating regenerated shoot tips. In this research we reported synthetic seed production and subsequent conversion of encapsulated shoot tips into plantlets comparing with nonencapsulated shoot tips for myrtle. Two different myrtle genotypes were used for synthetic seed production. Sodium alginate solution at the rate of 3.0% and 100?mM calcium chloride solution were prepared for encapsulation. Encapsulation was accomplished by mixing shoot tips into sodium alginate solution and dropping these in calcium chloride solution for 25–30?min. Encapsulated and nonencapsulated shoot tips were cultured in MS (Murashige and Skoog) media supplemented with different BAP (6-Benzylaminopurine) concentrations (0, 0.5, 1, 2?mg L^?1). After six weeks, all shoots were transferred to MS media containing 1?mg L^?1 IBA for in vitro rooting. As a result, the highest germination rate was obtained on the BAP-free MS media. The best BAP concentrations were detected as 0.5 and 1?mg L^?1 for micropropagation. Genetic stability of plants coming from encapsulated and nonencapsulated shoot tips was tested by ISSR markers. Based on the results, there were no genetic differences among the samples.

     

Development of a medium for in vitro culture of Galanthus species based on the mineral composition of bulbs

Summary

To optimise conditions for micropropagating Galanthus species, a basal medium (G) was developed based on mineral analyses of G. nivalis, G. nivalis ‘Flore Pleno’ and G. elwesii bulbs. Compared with Murashige and Skoog (MS) medium, the main features of G medium were increased concentrations of Cu ( 30.4), P ( 3.6), Ca ( 1.9), Mg ( 1.3) and S ( 1.2) and reduced levels of Mn ( 0.07), Zn ( 0.59) and K ( 0.65). The efficacy of G medium in supporting bulblet initiation on bulb chip explants, bulblet multiplication (on media supplemented with 30 g l^–1 sucrose, 1.0 mg l^–1 6-benzylaminopurine and 0.1 mg l^–1 naphthalene acetic acid), and bulblet growth (on plant growth regulator-free media with 60 g l^–1 sucrose and 5 g l^–1 activated charcoal) was compared with MS medium over a range of dilutions (full-, ¹?2-, ¹?4-, and ¹?8-strength). Bulblet initiation was superior on G medium for G. nivalis and G. nivalis ‘Flore Pleno’, but inferior for G. elwesii. The choice of basal medium did not influence bulblet multiplication, although multiplication was reduced on both media diluted to ¹?8-strength. G medium supported bulblet growth and rooting better than MS medium, while dilution of either medium reduced bulblet growth and rooting. Using G medium in place of MS medium during bulblet multiplication greatly reduced hyperhydration with G. elwesii, as did dilution of either of the basal media.     

Effect of agar,galactomannan and indole-butyric acid on in vitro rooting of the pear cultivar ‘Durondeau’ and apple rootstock cultivar ‘Marubakaido’

Summary

The influence of partial substitution of agar by galactomannans in culture media supplemented with different concentrations of indole-3-butyric acid (IBA) was studied on in vitro rooting of pear (Pyrus communis L.) cultivar ‘Durondeau’ and apple rootstock (Malus prunifolia Borkh.) cultivar ‘Marubakaido’. The galactomannans applied were obtained from Cassia fastuosa (cassia) and Cyamopsis tetragonolobus (guar gum) seeds. The results obtained with mixtures of agar and galactomannan (3 g l^–1 each) were compared with those from media solidified with a standard concentration of agar (6 g l^–1). The rooting of pear shoots was enhanced significantly in the presence of a mixture of agar plus cassia galactomannan compared to medium solidified with agar only. The modified media promoted a higher number of roots than the control, and increased the percentage of rooted shoots. A maximum of 84.8% rooting was obtained on half-strength MS medium (¹?2MS) supplemented with 0.49 µM IBA and solidified with a blend of agar plus cassia galactomannan. For the apple rootstock, only the number of roots per shoot was influenced significantly by the addition of galactomannan to the rooting medium. The highest number of roots per shoot was 16.67 on ¹?2MS medium gelled with a mixture of agar plus guar galactomannan supplemented with 4.90 µM IBA. The behaviour of the agar-galactomannan gel and the possibility of reduced costs when compared with systems containing only agar, suggest new biological and commercial applications for galactomannans.     

Rapid propagation of Cucurbita pepo L. by culture of meristem tips

A tissue culture technique has been developed for the rapid multiplication of pumpkin (Cucurbita pepo L.) clones. Meristem-tips from seedlings of cultivar ‘Cinderella’ were grown initially on MS medium containing 2.56 mg l^?1 Kinetin and 8 mg l^?1 IAA, and then transferred to experimental media. Maximum shoot proliferation occurred on MS medium containing 1 mg l^?1 BA and no auxin. Cultures were rooted after 2–3 weeks on MS medium containing 8 mg l^?1 IAA and no cytokinin.     

Rapid in vitro germination of zygotic embryos of fluted pumpkin (Telfairia occidentalis Hook. f.)

Summary

Fluted pumpkin, Telfairia occidentalis, is becoming an important regional vegetable for its food and medicinal uses. The recalcitrant nature of its seed makes conservation difficult and in vitro techniques may be a viable option for conservation. A pilot study was conducted on the effects of different concentrations of a commercial bleach [3.85% (w/v) sodium hypochlorite] for surface sterilisation of T. occidentalis seed. The optimum concentration [25% (v/v)] was then used as a basis to investigate the responses of mature embryonic axes of T. occidentalis to different concentrations of kinetin (Kin; 0, 1.0, or 2.0 mg l^–1) and 1-naphthaleneacetic acid (NAA; 0, 0.5, or 1.0 mg l^–1) combined in a factorial design. The results of the first experiments indicated that commercial bleach at 25% (v/v) resulted in the lowest contamination of explants (10%), with no evident injury to the embryonic axes. The results revealed that root emergence started 3 d after initiation (DAI) only on Murashige and Skoog medium (MS) with no added plant growth regulator (PGR), and that, by 12 DAI, all media supported the rooting of explants. The highest rooting percentage (69%) was observed at 15 DAI on MS medium with 0.5 mg l^–1 NAA, without Kin. However, shoot emergence started at 9 DAI on PGR-free MS medium, on MS with 0.5 mg l^–1 NAA, or on MS plus 1.0 mg l^–1 Kin. The highest shooting percentage (91%) of explants was observed with 0.5 mg l^–1 NAA at 21 DAI. Considering all other growth parameters, MS medium supplemented with 0.5 mg l^–1 NAA was found to be best for the germination of embryonic axes of T. occidentalis.     

Callus and axillary-bud culture of Vitis vinifera ‘Sylvaner Riesling’

Healthy growth of serially subcultured callus of the grape Vitis vinifera cultivar ‘Sylvaner’ was obtained by incubation at 30° C in continuous light in a defined culture medium containing 2% w/v sucrose, 1.0 mg l^?1 1-naphthaleneacetic acid (NAA) and 0.2 mg l^?1 kinetin (K). Organogenesis was not induced in this callus by alteration in the absolute or relative levels of NAA and K.Continued shoot initiation was obtained by culture of axillary buds in a medium containing 10^?5 M Benzyladenine (BA). Plantlets could be generated from these shoot buds by transfer to media containing 10^?7 M BA or lacking a cytokinin.     

Influence of medium-nitrogen level on growth periodicity of Ilex crenata Thunb. ‘Helleri’

Ilex crenata Thunb. ‘Helleri’ plants were grown in sand culture and supplied daily with nutrient solutions of 10, 40, 70 or 100 mg l^?1 nitrogen (N). Plant growth at all rates of N was characterized by an initial period of simultaneous shoot and root dry weight accumulation, followed by shoot elongation, root growth cessation, and major accumulations of N and dry weight. Plants grown at the greater N rates accumulated more N and exhibited a higher shoot-to-root ratio as a result of greater shoot than root growth. Plants grown at the higher N applications initiated extension growth before plants grown at lower N levels.     

Progress towards clonal propagation of Camellia japonica cv. Alba Plena by tissue culture techniques

Summary

Optimal rooting conditions have been determined for shoot cultures of Camellia japonica cv. Alba Plena derived from a 50 year old tree: dipping the base of shoots in 1 g l^?l solution for 15 min, followed by 12 days’ darkness, induced 87% rooting in shoots cultured with Woody Plant Medium (WPM) macronutrients. Halving the auxin concentration or exposure time considerably reduced this rate, and root formation was severely inhibited if an initial dark period for the entire shoot was not used. The type of support (agar or paper bridges) did not significantly affect the rooting percentage or number of roots per rooted shoot, but liquid media induced greater root elongation. No significant differences were observed between the use of WPM and a modified Heller’s medium as regards the rooting percentages achieved, but the number of roots formed was considerably greater with WPM, as was the survival rate after transfer to soil (70% for transfer as against 35% with the modified Heller’s medium). Best survival rates were achieved when shoots were transferred to the acclimatization soil 12 days after auxin treatment, i.e. immediately after the dark period.     

Effects of growth regulators and activated charcoal on in vitro bulblet multiplication and growth in Galanthus nivalis ‘Flore Pleno’

Summary

The in vitro multiplication of Galanthus nivalis ‘Flore Pleno’ bulblet clumps was evaluated through three 16-week sub-culture passages on media supplemented with either 1.0 mg l^–1 6-benzylaminopurine (BA) and 0.1 mg l^–1 naphthaleneacetic acid [NAA; the plant growth regulator (PGR) control], 0.5 mg l^–1 BA and 0.05 mg l^–1 NAA (PGR/2), or 0.1 mg l^–1 BA and 0.01 mg l^–1 NAA (PGR/10). At the end of the second sub-culture passage, clumps of bulblets from each of the three PGR treatments were also transferred to conditions to promote bulblet growth (G medium without PGRs) supplemented with 60 g l^–1 sucrose and 5 g l^–1 activated charcoal (AC). Lowering the PGR concentration during the initial multiplication phase reduced bulblet multiplication and overall culture growth, but did not influence bulblet size, as indicated by the diameter of the largest bulblet. Lowering the PGR concentration during the multiplication phase also reduced the multiplication of bulblets, overall tissue fresh weight, and the production of roots in tissues transferred to bulblet growth conditions. The reduced growth of tissues previously multiplied on PGR/10 medium was not, however, reflected in the growth of individual bulblets, since these were not significantly smaller than bulblets initially multiplied at the higher PGR concentrations. Bulblet sprouting and root growth on the bulblet growth medium were unaffected by prior PGR status in the multiplication phase. The results are discussed in terms of the mode of action of AC on the promotion of bulblet growth.     

Vegetative growth and nutritional status as influenced by auxins and gibberellic acid,and their effect on fruit yield in lemon

Application of 2,4-D and 2,4,5-T at concentrations ranging from 5 to 20 mg l^?1 to 5-year-old ‘Pant Lemon-1’ (Citrus limon Burm) trees reduced the vegetative growth in terms of height, spread, shoot length, number and size of the leaves in the autumn flush. Various NAA treatments (5–20 mg l^?1), however, enhanced growth, but not to the extent that was observed after GA₃ treatments. Application of GA₃ at 10–40 mg l^?1 significantly enhanced all aspects of growth, and the effects were most pronounced at 20 and 40 mg l^?1. Nutritional status of the leaves showed a slight variation in relation to vegetative growth under various treatments.Some 2,4-D- and 2,4,5-T-treatments increased the fruit yield over the control, which could suggest mobilization of foods even at the expense of reduced vegetative growth. On the other hand, NAA, particularly at 10 mg l^?1, increased both vegetative growth and yield, suggesting that the transport of the photosynthates from the leaves to the fruits was not at the expense of new growth extension. Due to excessive growth enhancement under higher concentrations of GA₃ (20 and 40 mg l^?1), comparatively fewer nutrients were translocated to the fruit “sinks”, thereby resulting in a non-significant decrease in yield.     

Micropropagation of red raspberry and the influence of phloroglucinol

Micropropagation of 12 raspberry seedling selections and the cultivar ‘Malling Jewel’ has been achieved. A basic culture medium (Linsmaier and Skoog, 1965) supplemented with benzylaminopurine (BAP), 1.0 mg l^?1, and indol-3-yl butyric acid (IBA), 0.1 mg l^?1, was optimal for shoot proliferation. The presence of phloroglucinol (PG) at a concentration of 162 mg l^?1 significantly increased shoot number at all auxin: cytokinin concentrations. Removal of the cytokinin and increasing the concentration of IBA to 1.0 mg l^?1 resulted in adventitious root formation. PG synergistically promoted the number of roots per rooted culture but did not significantly increase the percentage rooting. Viable plants were produced from all genotypes when transplanted to soil.     

Evaluation of the activity of aqueous extracts of some organic waste materials on in vitro-cultured shoots of ‘M9’ apple rootstock

Summary

The present work investigated the effects of different aqueous extracts of organic waste compounds on growth, proliferation and photosynthetic activity in ‘M9’ (Malus domestica Borkh.) shoot cultures, with the aim of determining the feasibility of using in vitro cultures as a tool for the rapid evaluation of organic amendments in agriculture. Aqueous extracts of the following organic waste compounds: cow manure (CM), sugarbeet industrial waste (SB), mixed grape, poultry and municipal solid waste (GPM), and citrus pruning and industrial waste (CPI) were prepared at a rate of 1:10 (w/v) compound:distilled water. The basal media used in the proliferation phase were: (i) PM1, modified Murashige and Skoog (MS) enriched with 4.4 µM 6-benzyladenine (BA); (ii) PM2, as PM1 but with a reduced cytokinin concentration (1 µM BA) to evaluate possible hormone effects; and (iii) PM3, 4.4 µM BA with reduced salt strength (0.33 MS) to induce nutrient deficiency. Hormone-free medium with half-strength MS salts was used for rooting. All media were enriched with each extract at 0, 0.2, 2, 20 or 200 ml l^–1. Photosynthetic activity was measured on PM3 medium enriched with SB or CM. Standard culture conditions were 22° ± 2°C, with a 16 h photoperiod at 30 µmoles photosynthetically active radiation (PAR) m^–2 s^–1, but at 80 µmoles PAR m^–2 s^–1 to determine photosynthetic activity. Shoot weight increase in PM1 was not affected by the GPM and CPI extracts, while the growth trends of CM- and SB-treated shoots were described by a second-degree function with maxima at 2 ml l^–1 and 0.2 ml l^–1, respectively. Shoot proliferation for SB was represented by a quadratic curve (maximum at 2 ml l^–1), was linearly reduced as GPM increased, but was not affected by CM or CPI. Treatments did not significantly affect rooting percentage and root length; however root number was increased by SB at 2 ml l^–1.CO₂ fixation increased linearly with both SB and CM, despite reduced growth at the highest levels of extract.     

Rapid micropropagation of Psoralea corylifolia L. using nodal explants cultured in organic additive-supplemented medium

Summary

The influence of different growth regulators and additives on shoot multiplication from nodal explants of Psoralea corylifolia was investigated. Prolific shoot multiplication was achieved within 4 weeks of culture on Murashige and Skoog (MS) medium supplemented with 5 μM benzyladenine (BA), 5 μM ascorbic acid (AA), 100 mg l^–1 casein hydrolysate (CH) and 5% (v/v) coconut water (CW). Shoots elongated on half-strength MS basal medium devoid of inositol, but containing 5 μM 2-isopentenyladenine (2iP), 10 g l^–1 sucrose and 8 g l^–1 agar. Elongated shoots rooted on half-strength MS basal medium supplemented with 3 μM indole-3-butyric acid (IBA), 10 g l^–1 sucrose and 7 g l^–1 agar within 5 d of culture. The in vitro-raised plants were established successfully in 2:1:1 (v/v/v) garden soil:farmyard soil:sand, and maintained in a growth chamber with 100% survival. Acclimatised plants were transferred to a glasshouse and established successfully in the field. Flowers and fruits appeared after 4 months and resembled those on source plants. This system could be used for rapid commercial propagation of P. corylifolia for conservation strategies and to produce phytomedicines.     

High-frequency embryogenesis,regeneration of broccoli (Brassica oleracea var. italica) and analysis of genetic stability by RAPD

High-frequency somatic embryogenesis and shoot regeneration of broccoli (Brassica oleracea var. italica) were achieved. Cotyledon and hypocotyl explants from four varieties of broccoli were cultured on MS and modified MS media (mMS, supplemented with PG-96 organic components) with different combinations of growth regulator. The effects of genotypes, different explants, growth regulator combinations, organic components and AgNO₃ on induction of calli and shoots were evaluated. The optimal media for inducting calli/shoots and roots were mMS medium containing 3% (w/v) sucrose and 0.8% (w/v) agar supplemented with NAA at 0.5 mg l⁻¹, 6-BA at 3.0 mg l⁻¹, AgNO₃ at 4.0 mg l⁻¹ and MS medium containing 3% sucrose and 0.8% (w/v) agar supplemented with NAA at 0.2 mg l⁻¹, respectively. The callus induction percentages were over 90% in all four varieties; shoot induction percentage was 92.5% and the average number of shoot per explant was 4.1 from cotyledon explant in variety Bishan. In this study, we established high-efficient embryogenesis and shoot regeneration system of broccoli and analyzed genetic stability of regenerants at DNA level using RAPD molecular marker. Out of 62 arbitrary primers screened using PCR amplification, 79 polymorphic bands were amplified from 20 primers. The results demonstrated the genetic stability of regenerants from the same variety.     

Rejuvenation influences indirect organogenesis from leaf explants of Pomegranate (Punica granatum L.) ‘Kandhari Kabuli’

Sequential subculturing leads to a gradual physiological change in cells that may be termed ‘rejuvenation’. The effect of repetitive subculturing on callus induction and shoot regeneration from leaf explants of Punica granatum L. ‘Kandhari Kabuli’ were investigated. Surface-sterilised leaves were cultured on 1.0× Murashige and Skoog (MS) medium supplemented with 4.0 mg l^–1 α-naphthaleneacetic acid (NAA) and 2.0 mg l^–1 6-benzyladenine (BA) for callus induction. Shoots were regenerated from callus on 1.0× MS medium supplemented with 1.5 mg l^–1 BA, 0.5 mg l^–1 kinetin, and 0.25 mg l^–1 NAA. Subculturing of callus onto fresh medium maintained the rate of shoot formation and substantially increased the production of shoot buds up to the second subculture. Following further subculture passages, a lower shoot regeneration potential from callus was observed. A maximum shoot bud induction from callus of 63.9% was observed at the second subculture passage. The rate of multiplication of in vitro shoots increased until the fourth subculture, then became constant. Similarly, in vitro rooting of micro-shoots increased up to the third subculture, followed by a decline during further subculturing.