Effect of nitrogen form and radiation on growth and mineral concentration of two Brassica species

Three greenhouse experiments were carried out to determine the growth, yield, nitrate, total N and S concentration in shoots, and water uptake of hydroponically grown Brassica rapa L. subsp. nipposinica var. chinoleifera and Brassica juncea L. In each experiment, daily photosynthetically active radiation (PAR) level was 5.0 mol m⁻² (low), 6.8 mol m⁻² (medium) or 9.0 mol m⁻² (high). Plants were supplied with nutrient solutions having equal N concentrations of 11 mM in different forms: 100% NH₄, 50% NH₄ + 50% NO₃, and 100% NO₃. Nitrogen supplied as 100% NH₄ reduced fresh and dry shoot biomass, leaf area, and leaf number in both Brassica species, especially at low and medium PAR levels. In both Brassica species, S concentrations were highest, while nitrate concentrations were lowest in leaves of plants grown at N supplied as 100% NH₄. No differences in leaf nitrate concentrations were observed between 50% NH₄ + 50% NO₃ and 100% NO₃ treatments. Low and high PAR levels increased the nitrate concentrations and decreased the N/S ratio in leaves of both crops compared to medium PAR level. Fresh shoot biomass was maximized in Brassica rapa when PAR level was above the medium value and nitrate was supplied in the nutrient solution as NO₃ or as a mixture of 50% NO₃ and 50% NH₄. The highest fresh shoot biomass of Brassica juncea was observed in all nutrient solution treatments at high PAR level.     

Plant regeneration of pansy (Viola wittrockiana) ‘Caidie’ via petiole-derived callus

An in vitro plant regeneration protocol for pansy (Viola wittrockiana) cultivar ‘Caidie’ from petioles was established as following: callus induction on a half-strength MS medium supplemented with 0.45 μmol l⁻¹ 2,4-d plus 8.9 μmol l⁻¹ BA, callus subculture on medium F (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 0.44 μmol l⁻¹ BA) and then on medium T (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 2.2 μmol l⁻¹ BA), shoot regeneration on medium D3 (MS media supplemented with 2.9 μmol l⁻¹GA₃, 23.6 μmol l⁻¹ AgNO₃, 0.02% active charcoal and 4.5 μmol l⁻¹ TDZ), shoot multiplication on medium M (half-strength MS medium containing NAA 1.1 μmol l⁻¹, TDZ 9.1 μmol l⁻¹ and GA₃ 8.7 μmol l⁻¹), and then shoot elongation and rooting on medium R (MS medium supplemented with 1.1 μmol l⁻¹ NAA and 1.1 μmol l⁻¹ BA). Subculture on appropriate medium was found to be important for successful shoot regeneration.     

Recurrent somatic embryogenesis and plant regeneration in Coriandrum sativum L.

An efficient method of repetitive somatic embryogenesis and plant regeneration was established in Coriandrum sativum L. Embryogenic callus was induced from cotyledon and hypocotyl segments on Murashige and Skoog (MS) medium with 4.52 μM 2,4-dichlorophenoxy acetic acid (2,4-D), upon subculturing on medium having same level of 2,4-D at an interval of 3 weeks developed somatic embryos, which progressed to cotyledonary stage through early developmental stages of somatic embryogenesis. The transfer of somatic embryos at an early cotyledonary and cotyledonary stage in clumps in succession to fresh 4.52 μM 2,4-D supplemented medium developed embryos in a cyclic manner. Upon transferal to embryogenic clumps (cotyledonary embryos) to modified MS medium (4 g l⁻¹ KNO₃, 0.29 g l⁻¹ NH₄NO₃, 3 mg l⁻¹ thiamine HCl, 0.5 mg l⁻¹ pyridoxine HCl, and 5 mg l⁻¹ nicotinic acid), the embryos irrespective of the cycles underwent maturation and germination. Germinating embryos transferred to half-strength MS medium favored healthy growth of plantlets. The system of recurrent somatic embryogenesis in coriander offers a system for genes transfer and also scale-up production of modified plants.     

Responses of cucumber plant to NH4 and NO3 nutrition: The relative addition rate technique vs. cultivation at constant nitrogen concentration

Different N sources (NO₃⁻, NH₄⁺, or NH₄NO₃) at different relative addition rates (RAR) were supplied to cucumber (Cucumis sativus L.), a species sensitive to NH₄⁺ toxicity. For comparison, cucumber plants were also grown at constant concentrations of 1 and 5 mM NH₄⁺ or 5 mM NO₃⁻. The fresh weight of NH₄⁺-fed plants at RAR 0.15 and RAR 0.25 day⁻¹ was similar to that of NO₃⁻-fed plants, while at RAR 0.35 or RAR 0.45 day⁻¹ growth reduction occurred. When available as a constant concentration, NH₄⁺ decreased plant growth at 5 mM. It is concluded that at low rates of N supply the relative addition rate technique can be used for growing cucumber plants with NH₄⁺ as sole N source without deleterious effects.     

Production of protocorm-like bodies with bioreactor and regeneration in vitro of Oncidium ‘Sugar Sweet’

To produce mass propagules of Oncidium ‘Sugar Sweet’, we tested the feasibility of producing protocorm-like bodies (PLBs) using 5 l balloon type air-lift bioreactors, and selected an optimal culture medium for shooting and rooting in vitro. The results showed that liquid bioreactor cultures were more efficient for PLBs proliferation when compared to solid- and liquid-agitated flask cultures. The maximum PLBs biomass (326.3 g per bioreactor, FW) was obtained in the immersion bioreactor culture, with the growth ratio reaching 10.9 after 50 days of culture. This was obviously higher than the ebb and flood bioreactor culture. During bioreactor culture, sucrose and electric conductivity (EC) in the culture medium were negatively correlated with PLBs growth; the highest PLBs fresh and dry biomass was obtained 40 days after culture. An inoculation density of 20 g (FW) was optimal for PLBs growth in a 3 l working volume of 5 l bioreactor. Furthermore, MS medium supplemented with 2.0 mg l⁻¹ BA for shooting and 0.5 mg l⁻¹ IBA for rooting was optimal during in vitro culture, the plantlets were successfully established in the potting substrate.     

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.     

不同铵硝配比对芥蓝伤流液组分及植株氮磷钾积累的影响

采用伤流液组分分析和植物营养分析的方法,探讨了不同铵硝配比（CK,0∶100;T1,10∶90;T2,25∶75;T3,50∶50）影响芥蓝生长的生理机制。结果表明：与全硝态氮营养液对照相比,3 个营养液增铵处理均提高了芥蓝植株的生物量和伤流强度,其中营养液增铵25%（T2）处理的效果最好。T1 处理芥蓝植株伤流液中蛋白态氮和P 的流量与CK 相比显著增加;T2 处理芥蓝植株伤流液中NO₃^--N、P、K 流量均为最高;T3 处理芥蓝植株伤流液中NH₄⁺-N、氨基酸态氮和蛋
白态氮流量最高,而NO₃^--N 和K 流量则显著低于CK。T1 处理和T2 处理显著提高了芥蓝植株N、P、K 的含量和积累量;T3 处理亦显著提高了芥蓝植株N、P 的含量和积累量,但显著降低了K 的含量和积累量。综合来看,营养液适量增铵（25%）处理的效果最好,可以提高芥蓝根系吸收能力,促进氮代谢,增加植株对N、P、K 的吸收和积累,从而促进植株生长。     

High frequency plantlet regeneration from callus and artificial seed production of rock plant Pogonatherum paniceum (Lam.) Hack. (Poaceae)

Pogonatherum paniceum (Lam.) Hack. is a rock plant with good potential for vegetative recovery on naked lands. A high frequency in vitro regeneration system was developed for P. paniceum. Calli were induced from explants of mature seeds, seedlings, young leaves, and stem segments on Murashige and Skoog (MS) medium supplemented with 1.0 mg L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.0 mg L⁻¹ α-naphthalene acetic acid (NAA) and 0.2 mg L⁻¹ 6-benzylaminopurine (BAP). High induction rates (59.57%) and regeneration rates (100%) were obtained from mature seed explants; calli were sub-cultured for over 2 years and still retained a high regenerative capacity. One seed explant resulted in 69,997 plants in 1 year. Shoot buds derived from calli were used for encapsulation in liquid MS medium containing 3% sucrose and two different alginate matrices (3% sodium alginate (w/v) + MS medium containing 3% sucrose and 3% sodium alginate + 1% activated carbon (w/v) + MS medium containing 3% sucrose) with a 20-min exposure to 2% CaCl₂ and 0.3% bavistin (w/v). The capsule with 3.0% sodium alginate (w/v) and 1% activated carbon (w/v) showed a higher conversion rate (61.58%) and stronger plantlets under non-aseptic conditions. These systems are useful for the rapid clonal propagation and dissemination of artificial seed material of P. paniceum for eco-recovery.     

The effect of various nitrogen fertilization and foliar nutrition regimes on the concentrations of nitrates,ammonium ions,dry matter and N-total in carrot (Daucus carota L.) roots

Two field experiments (Experiment I in 2003–2005 and Experiment II in 2004–2005) with carrot c.v. ‘Kazan F₁’ were conducted at Trzciana village (50°06′N, 21°85′E). The experiments were arranged in a split-plot design with four replications. Two sub-blocks were identified in both experiments: I, without foliar nutrition; II, receiving plant foliar nutrition. The plants were sprayed three times alternately with: 2% urea solution, 1% solution of multi-component ‘Supervit R’ fertilizer (produced by Intermag, Poland) and again with 2% urea solution. Combinations with diversified nitrogen fertilization were distinguished within both sub-blocks. Experiment I comprised of: (1) Control, (2) Ca(NO₃)₂ 70, (3) Ca(NO₃)₂ 70 + 70, (4) (NH₄)₂SO₄ 70 and (5) (NH₄)₂SO₄ 70 + 70. Experiment II included: (1) Control, (2) ENTEC-26 35 + 35, (3) ENTEC-26 70 + 70, (4) ENTEC 26 105 + 105, (5) NH₄NO₃ 35 + 35, (6) NH₄NO₃ 70 + 70, (7) NH₄NO₃ 105 + 105. Where 70 kg N ha⁻¹ was used before sowing, whereas 35 + 35, 70 + 70 and 105 + 105 kg N ha⁻¹ were applied before sowing and as top dressing. Solid nitrogen fertilizer was added to the soil (produced by): Ca(NO₃)₂, Yara International ASA (Hydro); (NH₄)₂SO₄, Zak?ady Azotowe in Tarnów, Poland; NH₄NO₃, Zak?ady Azotowe in Pu?awy, Poland; and ENTEC-26, COMPO GmbH & Co. KG, Germany. The research aimed at determining the effect of diversified nitrogen fertilization and foliar nutrition on NO₃⁻, NH₄⁺, N-total and dry matter (d.m.) concentrations in carrot, and N uptake by storage roots. In Experiment I, nitrogen fertilization did not affect NO₃⁻ concentration, whereas in Experiment II, the applied N treatment increased NO₃⁻ concentration in carrot in relation to the control, except for the storage roots of plants fertilized with ENTEC-26 35 + 35. Nitrogen fertilization applied in both experiments caused a significant increase in N-total concentration in carrot and N uptake by storage roots in comparison with the control plants. In both experiments, nitrogen fertilization had a different effect on the concentrations of NH₄⁺ and d.m. in carrot. What is more, foliar nutrition treatments in both experiments had a different effect on the concentrations on NO₃⁻, N-total, d.m. in carrot and N uptake by carrot storage roots.     

In vitro studies of controlling tomato puffiness enhanced by temperature,nitrogen source and sucrose concetration

Tomato puffiness, which occurs after auxin spray under field conditions, was investigated by using the in vitro fruit-culture technique. Tomato flowers were cultured in different media or at different temperatures for 3 days, to induce parthenocarpy, with 2-hydroxymethyl, 4-chlorophenoxyacetic acid (HCPA) and then parthenocarpic fruits were transferred to media free of HCPA.The higher the temperature during induction of parthenocarpy, the higher the degree of puffiness. Puffiness enhanced at 30°C was corrected by succinic acid-2, 2-dimethylhydrazide (SADH), which lowered the auxin level.High ratio of NO₃-N : NH₄-N in the medium increased the degree of puffiness. Puffiness was enhanced in the medium with NO₃-N alone as the nitrogen source was corrected by (SADH) and by [2-chloroethyl]trimethylammonium chloride (CCC). Different nitrogen levels with a constant ratio of NO₃-N to NH₄-N did not affect the degree of puffiness.High concentrations of sucrose (6 and 8%) in the medium considerably increased the degree of puffiness, but this could also be corrected by CCC and SADH.     

Restoration of rooting competence in a mature plant of Garcinia indica through serial shoot tip grafting in vitro

Apical and axillary buds from a high yielding, early fruiting elite tree (more than 20 years old) were cultured in woody plant medium (WPM) supplemented with 0.9 μM N⁶-benzyl adenine (BA). Multiple shoots were obtained on WPM basal medium containing 8.9 μM BA and 0.5 μM thidiazuron (TDZ). Elongation of axillary shoots was obtained in half-strength WPM medium supplemented with 0.4 μM BA. For root initiation, the elongated shoots were transferred to half strength WPM basal medium containing 2.5–245 μM indole-3-butyric acid (IBA) or 2.7–268.5 μM α-naphthaleneacetic acid (NAA) or the shoots were subjected to 2.5–53.9 mM IBA, 2.7–59.1 mM NAA dip for (30 s–30 min) and then transferred to half strength WPM basal medium. However, rooting was never achieved even after 2 months of culture.     

Evaluation of basal media for micropropagation of four highbush blueberry cultivars

Murashige and Skoog medium (MS), woody plant medium (WPM), and a mixture of equal parts of MS and WPM (MW) were compared for in vitro shoot proliferation and rooting of four highbush blueberry (Vaccinium spp.) cultivars. During the multiplication stage, the shoots on WPM showed worse growth than the other shoots. The MW produced the best shoot growth. The shoots on MS grew well but tended toward hyperhydricity in the multiplication stage, especially in the case of ‘Bluecrop’ shoots. Rooting was worst in the shoots multiplied on WPM, while the best rooting percentage of ‘Bluecrop’ shoots was obtained on MS, and that of ‘O’Neal’ on MW. The rooted shoots developed a good root system and were easily acclimatized after potting.     

In vitro multiplication of statice plantlets using sugar-free media

Clumps of statice (Limonium latifolium) plantlets grown photomixotrophically were used as explants and cultured for 25 days on a sugar-free modified Murashige and Skoog (MS) medium in Magenta-type vessels with the number of air exchanges of the vessel (NAE) being 3.8 h⁻¹, at a photosynthetic photon flux (PPF) of 100 μmol m⁻² s⁻¹ and a CO₂ concentration of 1500 μmol mol⁻¹ in the culture room. A factorial experiment was conducted with three levels of 6-benzylaminopurine (BA) concentration, namely 0, 0.25 and 0.5 mg L⁻¹, and two types of supporting material, agar and Florialite (a porous material). The control treatment was a photomixotrophic culture using a sugar- and BA (0.25 mg L⁻¹) containing agar medium in the vessel with NAE of 0.2 h⁻¹, at a PPF of 50 μmol m⁻² s⁻¹ and a CO₂ concentration of 400 μmol mol⁻¹ in the culture room. Leaf area, chlorophyll concentration and net photosynthetic rate were greater in the sugar-free medium treatment with a BA concentration of 0.25 mg L⁻¹ and Florialite than those in the control treatment. The number of shoots and dry weight per clump in the sugar-free medium treatment were comparable to those in the control treatment. Among the sugar-free medium treatments, the number of shoots increased with increasing BA concentration, however, the leaf area, dry weight, chlorophyll concentration and net photosynthetic rate decreased with increasing BA concentration. The use of Florialite significantly enhanced the growth and root induction as well as net photosynthetic rate, compared with the treatments that use agar. These results indicated that sugar-free medium micropropagation could be commercially applied to the multiplication of statice plantlets.     

Coconut water and BAP successfully replaced zeatin in olive (Olea europaea L.) micropropagation

The data presented report on trials conducted during 24 months using the Portuguese olive cultivar ‘Galega vulgar’. The effectiveness of coconut water, BAP, or kinetin, as possible zeatin substitutes in olive micropropagation protocols, was investigated. In all stages of the micropropagation process, the mineral and vitamin formulation of olive medium (OM) was used. Regarding culture establishment the best results were achieved when 50 ml l⁻¹ coconut water and 2.22 μM BAP were used as medium supplements. For the in vitro multiplication stage, the highest proliferation rates with an average of 3.4 new explants on each 30 days were achieved maintaining the coconut water concentration at 50 ml l⁻¹ and increasing BAP up to 8.87 μM. The effects of IBA and activated charcoal on the in vitro root induction were also studied. Rooting rates of over 85% were obtained by basal immersion of the explants in IBA solution at 3 g l⁻¹ for 10 s, followed by inoculation in the OM culture medium, added with 2 g l⁻¹ of activated charcoal and without growth regulators. All in vitro rooted plants were transferred into Jiffy-Pots filled with vermiculite–perlite 3:1 (v/v) substrate. Those were subsequently wetted with the OM mineral solution, placed into polystyrene plates each one with 100 Jiffy-Pots capacity, which were transferred to traditional rooting mist benches, on a water-cooling equipped greenhouse. Such a simple acclimatization procedure allowed for 95% of plants survival.     

Effects of N fertilizers and rates on yield,safety and nutrients in processing spinach genotypes

Two field experiments were carried out at the Experimental Field, Department of Food Science (TE, Italy) in 2004 and 2005 to evaluate the effects of genotypes, different N forms and N rates on yield, safety and nutritional features of processing spinach. Experiment 1, as treatments, included spinach genotypes and N forms (CO(NH₂)₂; Agricote; NH₄NO₃); experiment 2 included three N forms (Ca(NO₃)₂; (NH₄)₂SO₄; NH₄NO₃) applied at rates of 0, 75, 150, 200 kg N ha⁻¹. This research work confirmed differences among spinach genotypes in terms of efficiency in N use and oxalate and nitrate accumulation. Spinach accumulated much more nitrate in petioles and much more oxalate in blades indicating that nitrate and oxalate might play a counterrole to each other. Fertilizers containing N under forms not readily available to the crop, i.e. Agricote, CO(NH₂)₂ and (NH₄)₂SO₄, increased nitrate and oxalate accumulations less than fast N-release fertilizers, but their effect on yield was limited. Highest yield with contents of nitrate and oxalate lower than the limits imposed to avoid health problems, were achieved with Ca(NO₃)₂, at rates of 130 and 150 kg N ha⁻¹ NH₄NO₃.     

The effect of various nitrogen fertilization and foliar nutrition regimes on the concentrations of sugars,carotenoids and phenolic compounds in carrot (Daucus carota L.)

The aim of this research was to determine the influence of various forms, diverse doses, and dates of application of nitrogen fertilizers and foliar nutrition on the concentration of sugars, carotenoids and phenolics compound in carrot. Two field experiments (Experiment I in 2003–2005 and Experiment II in 2004–2005) with carrot ‘Kazan F₁’ were conducted in Trzciana (50°06′N; 21°85′E) in Poland. Both experiments were arranged in a split-plot design with four replications. Two sub-blocks were identified in both experiments: sub-block (A) without foliar nutrition and sub-block (B) with plant foliar nutrition. In sub-block (B), plants were sprayed three-times with: 2% (w/v) urea, a 1% (v/v) solution of multi-component ‘Supervit R’ fertilizer, and again with 2% (w/v) urea. Combinations with diversified nitrogen fertilization were distinguished within both sub-blocks. The treatments in Experiment I consisted of: (1) Control, (2) 70 kg N ha⁻¹ as Ca(NO₃)₂, (3) 70 + 70 kg N ha⁻¹ as Ca(NO₃)₂, (4) 70 kg N ha⁻¹ as (NH₄)₂SO₄ and (5) 70 + 70 kg N ha⁻¹ as (NH₄)₂SO₄, where 70 kg N ha⁻¹ was used preplant and 70 + 70 kg N ha⁻¹ was applied preplant and as a top dressing, respectively. The treatments in Experiment II consisted of: (1) Control, (2) 35 + 35 kg N ha⁻¹ as ENTEC-26, (3) 70 + 70 kg N ha⁻¹ as ENTEC-26, (4) 105 + 105 kg N ha⁻¹ as ENTEC-26, (5) 35 + 35 kg N ha⁻¹ as NH₄NO₃, (6) 70 + 70 kg N ha⁻¹ as NH₄NO₃, (7) 105 + 105 kg N ha⁻¹ as NH₄NO₃, where 35 + 35, 70 + 70, 105 + 105 kg N ha⁻¹ was applied preplant and as top dressing, respectively. Solid nitrogen fertilizer was added to the soil, as produced: Ca(NO₃)₂—Yara International ASA (Hydro), (NH₄)₂SO₄—Zak?ady Azotowe w Tarnowie, Poland, NH₄NO₃—Zak?ady Azotowe w Pu?awach, Poland and ENTEC-26–COMPO GmbH & Co., KG, Germany. In Experiment I, the highest sugar concentrations were found in carrot fertilized with (NH₄)₂SO₄ 70, while in Experiment II in the control and after fertilization with ENTEC-26 35 + 35 kg N ha⁻¹. In both experiments N-fertilization affected an increase in phenolic compound concentrations in comparison with the control. Experiment I revealed no significant effect of N-fertilization on carotenoid concentrations in carrot, however in Experiment II the highest concentration of these compounds was characteristic for the control plants and carrot fertilized with ENTEC-26 35 + 35. The foliar nutrition applied in Experiment I caused a decline in sugar concentration and an elevated carotenoid concentration, however it had no influence on the phenolic compound concentrations in carrot. Yet the foliar nutrition in Experiment II led to a decrease in phenolic and carotenoid compound concentrations, but it did not affect sugar concentration in carrot.     

Japanese holly growth and release pattern of ammonium and nitrate from controlled-release fertilizers

Applications of 0.85, 1.70 or 2.55 kg N m^?3 were made to Ilex crenata Thunb. Hetzi. Nitrogen sources included weekly applications of liquid fertilizer (NH₄NO₃) and single application of TVA's Sulfur coated urea (SCUT), Gold N (SCUG), Nitroform (NITR), isobutylidene diurea (IBDU), and an experimental N-containing Osmocote (OSCN). Shoot dry matter and leaf N from recently matured leaves were determined. Analyses of NH₄-N and NO₃-N were also conducted on the monthly leachates collected from the containers. Most controlled-release fertilizers (CRFs) produced greater dry weights at 1.70 kg N, while NH₄NO₃ produced more dry weight than CRF's at 0.85 or 2.55 kg N. Initially, for CRF's ammonium was more abundant than nitrate in the leachate; subsequently, nitrate became the dominant N form for most of the growing-season. All urea-based fertilizers had higher NH₄ levels in the leachate when compared with NH₄NO₃-based fertilizers.     

Effects of shading and NO3:NH4 ratio on the yield,quality and N metabolism in strawberry

The effect of 50% shading and NO₃:NH₄ ratio (0:100, 75:25, 50:50, and 25:75) in the nutrient solution on growth, yield, quality and N metabolism in hydroponically grown strawberry (Fragaria × ananassa var Camarosa) was evaluated. Both fresh and dry weights of leaves were significantly lower when a high concentration of either NO₃ (100%) or NH₄ (75%) was the sole N source in the nutrient solution. In unshaded plants, increasing of both NH₄ and NO₃ ratio in the nutrient solution reduced photosynthetic (Pn) rate, however in shaded plants the reduction of Pn became more pronounced at a higher ratio of NH₄ in the nutrient solution. The yield in terms of fresh and dry weight of fruit per plant was significantly increased at the 75:25 and 50:50 (NO₃:NH₄) treatments. Fruit size was significantly affected by the treatments, so that the biggest fruits in both shaded and unshaded plants were obtained under the 75:25 and 50:50 (NO₃:NH₄) treatments. Total soluble solid (TSS) in unshaded plants was increased with increasing NH₄ ratio in the nutrient solution, however in shaded plants it was reduced at high NH₄ ratio in the nutrient solution. In both shaded and unshaded plants, higher concentration of NH₄ significantly reduced the post-harvest life of the fruits. The increase of tissue N concentration was nearly proportional to the NH₄ concentration in the nutrient solution. The activity of nitrate reductase (NR) was increased by increasing NH₄ from 0 to 50% and then reduced at a higher ratio of NH₄ in the solution. Shading increased NH₄ concentration so that the shaded plant had nearly twice as high NH₄ concentration in the leaves. The increase of NH₄ concentration induced by shading could be partially the reduction of NH₄ assimilate because of the shortage of carbohydrate.     

Micropropagation of Vitis thunbergii Sieb. et Zucc., a medicinal herb,through high-frequency shoot tip culture

This study describes a protocol for rapid and large-scale in vitro propagation of the valuable medicinal herb Vitis thunbergii Sieb. et Zucc.. Culture conditions influencing shoot proliferation and rooting of the two clones (three- and five-lobed) were examined. Three medium formulations, Murashige-Skoog (MS), Woody Plant Medium (WPM) and Nitsch and Nitsch (NN) medium, were tested for growth of shoot tip culture, and WPM was found to have a superior proliferation rate. The chlorophyll content of leaves was highest in those cultured on NN, followed by those on MS and WPM medium. WPM medium supplemented with 0.5 mg l⁻¹ 6-benzyladenine (BA) displayed the highest proliferation rate (15–19 nodes or 3–4 shoots per explant). The rooting was optimized using MS medium supplemented with 0.5 mg l⁻¹ napthaleneacetic (NAA) with eight roots, 3 cm long after 1 month of culture. High frequency callus formation was observed in the basal end of explants cultured on NAA-containing medium. Following acclimatization, rooting plantlets were transferred to the plastic house with a 95% survival rate.     

High frequency early flowering from in vitro seedlings of Dendrobium nobile

Dendrobium nobile Lindl. is a popular temperate Chinese orchid commonly marketed as a traditional medicinal plant. Seedlings of Dendrobium nobile Lindl. produced floral buds (33.3–34.8%) precociously on a defined basal medium (1/2 MS) containing paclobutrazol (PP₃₃₃) at 0.5 mg L⁻¹ or thidiazuron (TDZ) at 0.1 mg L⁻¹ within 4 months of culturing. The frequency of floral buds formation can be further increased to 95.6% by growing seedlings in a PN (PP₃₃₃ 0.3 mg L⁻¹ + NAA 0.5 mg L⁻¹)-containing medium followed by transfer onto 1/2 MS medium with PP₃₃₃ and TDZ (PP₃₃₃ + TDZ). However, flower developed was deformed under 25 °C but it developed fully when grown in a lower temperature regime (23 °C/18 °C, light/dark) for 45 days. Under optimal condition, in vitro flowering was observed about 6 months after seed sowing.