Plant regeneration of Carica papaya L. through somatic embryogenesis in response to light quality,gelling agent and phloridzin

Difficulties to develop an easy and reproducible protocol to get healthy and well formed plants from somatic embryos of papaya (Carica papaya L.) had included low germination, callus production at the base of the embryo radicle and the occurrence of hyperhydric plantlets among others, and by consequence unsuccessful transfer to the field. With the aim of improving a propagation method, the effects of light quality, gelling agent and phloridzin concentration on the germination of somatic embryos of hermaphrodite C. papaya L. var. Maradol were studied. Somatic embryos were grown on half strength MS medium, with the addition of Chen vitamins [Chen, M.H., Wang, P.J., Maeda, E., 1987. Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants. Plant Cell Rep. 6, 348–351], solidified with three distinct gelling agents: Sigma^® Agar–Agar, Difco^® Bacto agar and Phytagel^®; supplemented with phloridzin and exposed to different light qualities: blue (54 μmol m⁻² s⁻¹), red (65 μmol m⁻² s⁻¹), gro-lux (68 μmol m⁻² s⁻¹), red + blue, white (32 μmol m⁻² s⁻¹) and wide spectrum (49 μmol m⁻² s⁻¹) during a period of 4 weeks. Results show that light quality and gelling agent had important effects on germination and plant growth, while 3.0 mg L⁻¹ phloridzin had an important role on germination as well as in root development. Somatic embryos exposed to white light, culture medium solidified with 3.0 mg L⁻¹ phytagel and 3.0 mg L⁻¹ phloridzin showed longer roots. Meanwhile, germination and plant length were promoted on an improved culture medium solidified with 7.5 g L⁻¹ Difco^® Bacto agar, 3.0 mg L⁻¹ phloridzin and exposed to gro-lux lamps. Under these conditions, 70% of somatic embryos germinated and developed normal roots without hyperhydricity. The regenerated plantlets with well developed roots and shoots were successfully transferred to a greenhouse with a survival rate of 95%.     

Comparison of ‘Verna’ lemon juice quality for new ingredients and food products

Two clonal selections of lemon trees (‘Verna-62’ and ‘Verna-50-2’), were studied in aiming to ascertain the influence of genetic (clone) and environmental (season) factors on the human-health bioactive compounds of the lemon juice (vitamin C and flavonoids) and the possible relationship between composition and in vitro antioxidant capacity (DPPH, ABTS and FRAP) of the juice. The average values determined in bioactives were 13–26 mg·100 mL⁻¹ in total of analysed phenolics and 23–29 mg·100 mL⁻¹ in vitamin C content. Variability in the weather conditions and internal physiological phenomena of the fruits such as biosynthesis and transport, could determine, at least in part, differences in the contents of lemon juice bioactives more importantly than the genetic background, providing the food industry with phytochemically rich and nutritive lemons for processing and functional ingredients.     

Priming abiotic factors for optimal hybrid Cymbidium (Orchidaceae) PLB and callus induction,plantlet formation,and their subsequent cytogenetic stability analysis

Abiotic factors affect the induction of PLBs and callus in hybrid Cymbidium Twilight Moon ‘Day Light’. The initiation and proliferation of new PLBs and callus could be achieved on NAA and kinetin, supplemented at 0.1 mg l⁻¹ each, respectively, both within 45–60 days. Bacto agar was found to be the most suitable solidifying agent for PLB induction, although a higher shoot fresh weight was obtained on Gelrite; a pH 5.3 was optimal while pH 4.5 caused 100% explant necrosis; coconut water, when supplied at 10–20% (v/v) resulted in a significant increase in the number of PLBs formed per PLB segment (23.1 versus 14.6 in controls) while a massive (almost four-fold) increase in fresh top weight occurred when PLB explants were placed in liquid culture, as a result of hyperhydricity; Fe-EDTA (1 mg l⁻¹) and activated charcoal (1 g l⁻¹) stimulated total fresh weight and PLB formation in the presence of PGRs; PLB formation decreased but total fresh shoot weight increased with the addition of niacin or myo-inositol, both vitamins. Dark-grown PLB-induced plants were etiolated and had longer internodes and higher fresh weight than light-grown control plants at 45 μmol m⁻² s⁻¹; at 15 μmol m⁻² s⁻¹ shoots were slightly etiolated, fragile, and PLB formation was scarce. RAPD and mtDNA analysis of all resultant PLBs, callus or plants showed them to be genetically identical, with comparable chlorophyll contents. Despite the detection of cytological variation between different plant parts, little variation resulted from abiotic factor treatment.     

Total phenolics and flavonoids and total antioxidant capacity in pistachio (Pistachia vera L.) nuts in relation to cultivars and storage conditions

The effects of cultivar, drying and storage conditions on total phenolics (TP), total flavonoids (TF) and total antioxidant capacity (TAC), measured with ferric reducing antioxidant power (FRAP) and radical scavenging capacity (2,2-diphenyl-1-picrylhydrazyl or DPPH) assays, of in-shell and unpeeled kernels of ripe pistachios were investigated. Drying was carried out at 45 °C for 34 h to approximately 5% moisture in kernels and storage of dried nuts in packaging atmosphere of dry air or N₂ at 1 °C or 20 °C for 6 or 12 months. At harvest, each cultivar showed its highest values for each measured variable and TP ranged from 16.2 to 7.9 mg gallic acid equiv. g⁻¹ d.w., TF from 7.2 to 3 mg catechin equiv. g⁻¹ d.w., FRAP from 132.5 to 58.8 μmol Trolox equiv. g⁻¹ d.w., and DPPH from 122.6 to 45.7 μmol Trolox equiv. g⁻¹ d.w. Drying resulted in losses in all cultivars that averaged approximately 14.2%, 14.1%, 11.9% and 12.2% for TP, TF, FRAP and DPPH, respectively, while during 12-month storage the corresponding losses in dried kernels averaged approximately 24.7%, 21.8%, 30.3% and 32.4%. Decreases in all measured variables were advanced by storage time, but prevented by low temperature and packaging in N₂ atmosphere. Among the studied cultivars, Pontikis, Aegina, Bronte and Cerasola showed higher values of TP, TF and TAC than Sirora, Kerman, Joley and Mumtaz in all cases, while Pontikis the highest in most cases. The effects of cultivar, time, temperature and packaging atmosphere during storage were all significant on TP, TF, FRAP and DPPH. Strong correlations were also found among the measured variables.     

Efficient in vitro multiplication in Ornithogalum ulophyllum Hand.-Mazz. from twin scale explants

Ornithogalum ulophyllum Hand.-Mazz. with beautiful white flowers is an important medicinal and ornamental plant of the Middle Eastern countries and need exploitation for commercial propagation. The study reports in vitro mass proliferation of bulblets achieved from twin scales and “in vitro regenerated bulblet” explants on MS medium supplemented with various concentrations of BAP–NAA. The best regeneration on twin scales and “in vitro regenerated bulblets” was obtained on MS medium containing 2 mg l⁻¹ BAP–0.5 mg l⁻¹ NAA and 2 mg l⁻¹BAP–1 mg l⁻¹ NAA, respectively. However, bulb scales seemed to be more potent for bulblet regeneration. A large number of the developing bulblets rooted on the regeneration medium. Remaining non-rooting bulblets were rooted on MS medium containing 1 mg l⁻¹ NAA. All plants were acclimatized in the environmental chamber for 4 weeks and were transferred to the greenhouse for flowering. Regenerated bulblets developed into morphologically normal plants.     

Quantification of phenolic acids and antioxidant activity in sweetpotato genotypes

Phenolic acid composition and antioxidant activity in roots of 14 commercially important sweetpotato genotypes were evaluated. Significant differences in total phenolics, individual phenolic acids, and antioxidant activity were found among the different sweetpotato genotypes. Total phenolic content, expressed in terms of chlorogenic acid equivalent, in different genotypes ranged from 1.4 to 4.7 mg g⁻¹ dry weight (DW). Antioxidant activity was evaluated as Trolox equivalent, ranging from 1.3 to 4.6 mg g⁻¹ DW. The highest total phenolic content and antioxidant activity were observed in a purple-fleshed genotype. Chlorogenic acid and 3,5-dicaffeoylquinic acid were the predominant phenolic acids, while caffeic acid was the least abundant in most genotypes. The highest content of chlorogenic acid (422.4 μg g⁻¹ DW) was present in a white-fleshed cultivar ‘Quarter Million’ imported from Jamaica. However, a purple-fleshed genotype had the highest amounts of 3,5-dicaffeoylquinic (485.6 μg g⁻¹ DW), 3,4-dicaffeoylquinic (125.6 μg g⁻¹ DW), 4,5-dicaffeoylquinic (284.4 μg g⁻¹ DW), and caffeic (20.5 μg g⁻¹ DW) acids.     

Micropropagation of the bromeliad Guzmania ‘Hilda’ via organogenesis and the effect of α-naphthaleneacetic acid on plantlet elongation

This study established a highly effective micropropagation system to obtain good plantlet proliferation from floral organs via callus induction and bud differentiation in Guzmania ‘Hilda’ bromeliad. The best frequencies of organogenic callus formation (20% in petal and 35% in ovary explants) were obtained on media containing a combination of 1.0 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) with 1.0 mg l⁻¹ α-naphthaleneacetic acid (NAA) and 1.5 mg l⁻¹ 2,4-D with 0.5 mg l⁻¹ NAA, respectively. Organogenic calli were cultured on medium with 1.0 mg l⁻¹ NAA and 0.5 mg l⁻¹ 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ) induce the differentiation and regeneration of adventitious buds into plantlets. When the plantlets were cultured in a medium with optimum NAA concentration (0.5–1.0 mg l⁻¹) significant improvement in regeneration and elongation was achieved within one month. This overcame the difficulty of delayed elongation in Guzmania plantlets. More than 99% of the regenerated and acclimatized plantlets developed to the flowering stage.     

Effect of ABA and sucrose on germination of encapsulated somatic embryos of guava (Psidium guajava L.)

Present study demonstrates the effect of sucrose and ABA on germination of encapsulated somatic embryos of guava (Psidium guajava L.). Sucrose and ABA at different concentrations were also evaluated for their effects on maturation and germination of somatic embryos. Mature somatic embryos developed on MS medium containing high concentration of sucrose (10%) or ABA (1.0 mg l⁻¹) showed inhibition in germination if they continued to be in same medium for 4 weeks. With increasing concentrations of sucrose (3–9%) or ABA (0.01–1.0 mg l⁻¹) in medium, percent germination of encapsulated somatic embryos decreased significantly. Encapsulated somatic embryos after storage on MS medium supplemented with 9% sucrose or 1 mg l⁻¹ ABA for different duration (0–60 days) germinated when they were transferred to medium containing 3% sucrose. About 20.8% and 37.5% encapsulated somatic embryos germinated after storage on ABA (1 mg l⁻¹) or sucrose (9%) for 60 days, respectively. Temporarily suppression in germination of encapsulated somatic embryos by high concentration of sucrose or ABA may be important for short-term conservation of elite genotype of guava.     

Investigation of physico-chemical properties and antioxidant activity of twenty Iranian pomegranate (Punica granatum L.) cultivars

Pomegranate is one of the native fruits of Iran which contains high genetic resources, but there are insufficient information regarding properties of the fruit. The objective of the present study was to investigate the physcio-chemical characteristics and antioxidant activity of twenty pomegranate cultivars grown in Iran. This study showed that there were significant differences among the cultivars in all measured factors except the length/diameter ratio of fruit. The fruit weight, skin percentage, aril percentage and juice percentage were within the range of 196.89–315.28 g, 32.28–59.82%, 37.59–65% and 26.95–46.55%, respectively. The total soluble solids content varied from 11.37 (°Brix) to 15.07 (°Brix), pH values from 3.16 to 4.09, titratable acidity content from 0.33 g 100 g⁻¹ to 2.44 g 100 g⁻¹ and total sugars content from 13.23 g 100 g⁻¹ to 21.72 g 100 g⁻¹. The results also showed that the values of ascorbic acid ranged from 9.91 mg 100 g⁻¹ to 20.92 mg 100 g⁻¹. The total anthocyanins content was observed in pomegranate cultivars between 5.56  mg 100 g⁻¹ and 30.11 mg 100 g⁻¹. The level of total phenolics was varied from 295.79 mg 100 g⁻¹ to 985.37 mg 100 g⁻¹. The antioxidant activity of pomegranate cultivars was found between 15.59 and 40.72%. These data demonstrated that the cultivar was the main parameter which influences the physico-chemical properties and antioxidant activity in pomegranates.     

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.     

Chromosome doubling of Lychnis spp. by in vitro spindle toxin treatment of nodal segments

Lychnis (Caryophyllaceae) consists of about 30 species distributed throughout the temperate regions of the Northern Hemisphere, from East Asia to Europe. Many Lychnis spp. have high ornamental value and cultivated as pot or garden plants. In the present study, in vitro chromosome doubling of several Lychnis spp. was examined in order to widen their variability in horticultural traits. Initially effect of various spindle toxin treatments [100, 500 or 1000 mg l⁻¹ colchicine (COL), 10, 20 or 50 mg l⁻¹ oryzalin (ORY), or 1, 5, 10 mg l⁻¹ amiprophos-methyl (APM)] of nodal segments of a triploid genotype of L. senno (3x) was investigated on survival of nodal segments and chromosome doubling in nodal segment-derived plantlets. Significantly higher percentage (75.0%) of surviving segments after spindle toxin treatment was obtained in 10 mg l⁻¹ ORY treatment. Flow cytometry (FCM) analysis of leaf tissues showed that 9.4–13.8% of plantlets, which were derived from 10 to 20 mg l⁻¹ ORY, or 5 mg l⁻¹ APM treatments, were hexaploid (6x) or ploidy chimera (3x + 6x, 4x + 6x, 5x + 6x, 3x + 4x + 6x). The results obtained by FCM analysis were confirmed by chromosome observation in root tip cells. Thus 10 mg l⁻¹ ORY treatment of nodal segments is suitable for in vitro chromosome doubling of triploid L. senno. Efficient chromosome doubling was also achieved in diploid L. fulgens (2x) and L. sieboldii (2x) by treating nodal segments with 10 mg l⁻¹ ORY: 68.9–88.7% of nodal segments survived after ORY treatment, and 24.7–26.5% of plantlets derived from ORY-treated nodal segments were tetraploid (4x) or ploidy chimera (2x + 4x) in both species. These results indicate that the in vitro chromosome doubling method established for triploid L. senno may be applicable to a wide range of Lychnis spp. Tetraploid L. fulgens and L. sieboldii showed a compact plant form, and had thick stems and deep green leaves compared with the diploid mother plants. On the other hand, hexaploid L. senno showed very poor growth and died before flowering.     

Effect of 2,4-d, glutamine and BAP on embryogenic suspension culture of date palm (Phoenix dactylifera L.)

The proliferation of embryogenic suspension culture in two cultivars (Jihel and Bousthami Noir) of Phoenix dactylifera L. was tested on liquid media with or without 2,4-d and with different glutamine concentrations (3.35 × 10⁻⁴, 6.7 × 10⁻⁴ and 13.4 × 10⁻⁴ M). The liquid medium with 0.1 mg l⁻¹ 2,4-d and 6.7 × 10⁻⁴ M glutamine has clearly improved the proliferation of somatic embryos. In fact, when glutamine concentration increased from 3.35 × 10⁻⁴ to 6.7 × 10⁻⁴ M, the yield of somatic embryos increased from 14 to 56 embryos per 100 ml of culture medium for “Jihel” cultivar and 25–71 embryos per 100 ml of culture medium for “Bousthami Noir” cultivar. In contrast, increasing glutamine concentration from 6.7 × 10⁻⁴ to 13.4 × 10⁻⁴ M, the embryos yield was negligible. Based on biochemical analysis, the highest accumulation of proteins and sugars was obtained in liquid medium with 0.1 mg l⁻¹ 2,4-d and 6.7 × 10⁻⁴ M glutamine (118 and 91 mg of proteins g⁻¹ DW, respectively, for “Jihel” and “Bousthami Noir” cultivars; 194 mg of sugars g⁻¹ DW for “Jihel” cultivar and 182 mg of sugars g⁻¹ DW for “Bousthami Noir” cultivar). In addition, the supply of 0.05 mg l⁻¹ BAP on the germination medium could be useful in terms of germination percentage of somatic embryos. When BAP concentration increased from 0.05 to 0.2 mg l⁻¹, the germination percentage of somatic embryos decreased from 14.2 to 4.9%, while secondary embryogenesis increased from 26.4 to 45.2%.     

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.     

A time-course study of flavonoids in the sprouts of tartary (Fagopyrum tataricum Gaertn.) buckwheats

The evolution, from 1 to 10 days after germination, of flavonoid content in sprouts of tartary buckwheat (Fagopyrum tataricum Gaertn.), grown in a greenhouse under low light conditions (16 μmol m⁻² s⁻¹), was investigated. Chlorogenic acid and flavonoids including C-glycosylflavones (orientin, isoorientin, vitexin, isovitexin), rutin and quercetin were separated from the sprouts by HPLC and quantified with their commercial standards. Rutin content in the edible portion of the sprouts (mean 20 and 37 mg g⁻¹ DW in ‘Hokkai T 8’ and ‘Hokkai T 10’, respectively) was 3- to 31-fold greater than that in the roots or pericarp. The free radical scavenging activity of seed sprouts was assessed through the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. From 6 to 10 days after sowing, the free radical scavenging activity in the edible portions rose significantly from 1.52 to 2.33 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 8’ and from 1.46 to 2.09 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 10’, but differences between ‘Hokkai T 8’ and ‘Hokkai T 10’ were not significant. As the results, the sprouts of tartary buckwheat, particularly those of ‘Hokkai T 10’ are strongly recommended as new high rutin food.     

Effects of exogenous application of plant growth regulators on the development of ovule and subsequent embryo rescue of stenospermic grape (Vitis vinifera L.)

Seedless grapevine cultivars (Vitis vinifera L.) are widely grown in Europe, America and Asia. Abortion of zygotic embryos in seedless grapes largely limits the efficiency of breeding of seedless cultivars through genetic crossing. The present study was designed to investigate effects of exogenously applied plant growth regulators (PGRs) to the grapevines in field condition on ovule and subsequent embryo rescue of seedless grapes of small seed traces. First experiment was performed by measuring ovules weight, proportion of each category ovules in maturity and embryo development in vitro of seedless grape cv. Centennial Seedless, Thompson Seedless and Crimson Seedless sprayed by chlormequat (CCC), benzyladenine (BA), ethephon (CEPA) and putrescine (Put). The effects of different application concentration and date of CCC were further evaluated in Centennial Seedless in later experiment. The results showed that exogenous application of all PGRs did not affect the total number of ovules per berries in maturity. CCC increased the ovules weight and proportion of ovules >4 mm in length of three varieties in maturity. The effects of two application times of PGRs on weight of berries and ovules and proportion of each category ovules in maturity were not significantly different. In the proceeding of embryo rescue, CCC at 100 and 1000 mg l⁻¹, BA at 100 mg l⁻¹ and Put at 20 mg l⁻¹ increased the percentage of developed embryos of Centennial Seedless and Thompson Seedless. The results showed that the size of ovules excised for embryo rescue significantly affected embryo formation and plant regeneration. The percentage of embryos formation in ovules >4 mm in length was significantly more than in ovules 2–4 mm in length, no embryo was found in ovules <2 mm in length. Exogenous application of CCC at 100–500 mg l⁻¹ significantly increased percentage of ovules >2 mm in length by 80.0–82.7% in Centennial Seedless, therefore improving embryo formation. The statistical correlation was found between the proportion of ovules >2 mm and embryo formation (r = 0.92) in Centennial Seedless. Among the different spraying time in Centennial Seedless, CCC applied 14 days before bloom produced significantly more ovules >2 mm in length and embryos formation.     

Direct regeneration of plants derived from in vitro cultured shoot tips and leaves of three Lysimachia species

The regenerability of three ornamental species—Lysimachia christinae, Lysimachia rubinervis and Lysimachia nummularia ‘Aurea’, were investigated using in vitro leaves and shoot tips. 6-Benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) added to Murashige and Skoog (MS) medium were tested for their effect on organogenesis. On the medium, shoot regeneration occurred directly without callus formation. In these species, L. christinae developed the highest regeneration rate and numbers of shoots/explant from shoot tips (100%, 12.25) and leaf bases (100%, 13.01) on the MS medium containing 3.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. For L. rubinervis, the highest shoot induction rate and number of shoots/explant were obtained from shoot tip (100%, 16.87–17.20) on the MS medium with 0.1 mg l⁻¹ NAA and 3.0–5.0 mg l⁻¹ BAP. L. nummularia ‘Aurea’, however, showed the highest regeneration rate and number of shoots/explant (100%, 12.73) from leaf bases on MS medium supplemented with 1.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. All in vitro shoots rooted well on half macronutrient MS medium containing 0.1 mg l⁻¹ NAA. After acclimatization, transplanted plantlets grew normally and flowered in the field.     

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.     

Seed germination and tissue culture of Cymbidium giganteum Wall. ex Lindl.

Efficient protocols were established for in vitro seed germination, neo-formation of secondary (2°) protocorms from primary (1°) protocorms and multiple shoot buds and protocorm-like body (PLB) induction from pseudo-stem segments of in vitro-raised seedlings of Cymbidium giganteum. Four nutrient media, namely Murashige and Skoog (MS), Phytamax (PM), Mitra et al. (M), and Knudson ‘C’ (KC) were evaluated for seed germination and early protocorm development. In addition, the effects of peptone, activated charcoal (AC) and two plant growth regulators [6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D)] were also studied. Both M and PM supplemented with 2.0 g l⁻¹ peptone or 1.0 mg l⁻¹ BAP resulted in ∼100% seed germination. Media supplemented with 2.0 g l⁻¹ AC could effectively induce large protocorms (1.6 ± 0.1 mm in diameter). Neo-formation of 2° protocorms from 1° protocorms was achieved in liquid and agar-solidified PM medium fortified with different concentrations and combinations of auxins (α-naphthalene acetic acid (NAA) and 2,4-D) and cytokinins [BAP and kinetin (KN)]. The highest number of 2° protocorms was obtained in liquid medium (10.7 ± 0.9/1° protocorm) supplemented with 2.0 mg l⁻¹ BAP + 1.0 mg l⁻¹ NAA. Although protocorms proliferated profusely in liquid medium, these did not develop further unless transferred to agar-solidified medium within 6–8 weeks. Multiple shoot buds and PLBs were induced from pseudo-stem segments on agar-solidified PM medium fortified with different concentrations and combinations of BAP and NAA and the maximum number of PLBs (6.00 ± 0.20) was recorded when BAP and NAA were applied at 2.0 mg l⁻¹ each. A solid root system was induced from PLBs and shoot buds when these were transferred to half-strength PM or M media fortified with 0.5 mg l⁻¹ indole-3-acetic acid. Well-rooted plants were transferred to the greenhouse with 95% survival.     

Late season harvest and storage of Rubus berries—Major antioxidant and sugar levels

Late season production of raspberries and blackberries has become more common due to increased consumer demand and new production techniques, but information is scarce regarding the effect on nutritional quality, especially during storage. Antioxidant levels in berries can be influenced by pre-harvest climate conditions (light intensity, day length and temperature) and post-harvest storage time. This study analysed the effect of these factors on the content of different antioxidants (vitamin C, ellagic acid, anthocyanins and total phenolics) in raspberries (Rubus idaeus L. cv. Polka) and blackberries (Rubus fructicosus L. cv. Loch Ness) during storage. The analyses were performed using high performance liquid chromatography (HPLC) with photodiode array detection. Ellagic acid content during storage ranged between 104 and 114 mg/100 g fresh weight in raspberries and 172 and 182 mg/100 g fresh weight in blackberries. Anthocyanin content increased during storage from 64 to 77 mg/100 g fresh weight in raspberries, but decreased from 199 to 162 mg/100 g fresh weight in blackberries. Vitamin C content in raspberries ranged during storage between 22 and 28 mg/100 g fresh weight, but in blackberries values were lower (10–12 mg/100 g fresh weight). Significant changes in total phenolics and sugars occurred during different storage periods (0–9 or 10 days). The results suggest that late season raspberries and blackberries maintain high quality in terms of antioxidant content.     

Physiological and biochemical changes with special reference to mangiferin and oxidative enzymes level in malformation resistant and susceptible cultivars of mango (Mangifera indica L.)

Changes in biophysical attributes, mangiferin and polyphenol oxidase (PPO), catalase and peroxidase activities in malformation resistant mango cultivar Elaichi were studied at various stages of flower development and compared with susceptible cvs. Amrapali, Beauty Mc-lin and Dashehari. Accumulation of mangiferin was maximum (96.0 and 108.0 mg g⁻¹ FW) in Elaichi prior to flower bud differentiation (September) and at full bloom (February), while these were minimum (59.0 and 74.0 mg g⁻¹ FW) in susceptible cv. Beauty Mc-lin. Mangiferin promoted vegetative growth and exhibited inhibitory role on the occurrence of malformation. It was also found that the resistant cultivar had highest activity of PPO as compared to susceptible ones. There was no significant difference in the enzymes catalase and peroxidase activity at early stage of flower differentiation but at flower bud burst stage the catalase activity was enhanced significantly in cv. Elaichi (25.28 unit min⁻¹ g⁻¹ FW) in comparison to Amrapali (16.20 unit min⁻¹ g⁻¹ FW), Beauty Mc-lin (18.39 unit min⁻¹ g⁻¹ FW) and Dashehari (17.50 unit min⁻¹ g⁻¹ FW). The resistant cultivar had high leaf temperature (30.30 °C) and diffusion resistance (476.14 m mol m⁻² s⁻¹) during the flowering but the rate of transpiration and relative humidity (RH) were high in susceptible cultivars. Results of the present study clearly indicate that level of mangiferin could be considered as a potential biochemical indicator for screening mango genotypes to malformation.