Heterosis for mineral elements in single cross-hybrids of cabbage (Brassica oleracea var. capitata L.)

Cabbage (Brassica oleracea var. capitata L.), a cross-pollinated crop, was observed to exhibit strong heterosis for high yield; better plant stand; early maturity; larger and more uniform heads; uniformity in head compactness; and disease tolerance in F₁ hybrids. However, information on the heterosis for mineral elements such as Fe, Zn, Cu, Mn, K and Ca although is not available to the best of our knowledge but it is important as it is likely to influence the plant and subsequently the human nutrition. Therefore, an attempt was made to estimate the heterosis for mineral elements in cabbage. Significant mean square for parents and hybrids was observed for all minerals under study which indicated the prevalence of sufficient variation. The parents 83-2, Pride of Asia, Red Cabbage, AC-204 and MR-1 were found to have the potential for use in cabbage quality breeding programme as they exhibited higher hybrid effects for Fe, Zn, Cu and Mn content. The single cross-hybrids, i.e. 83-2 × AC-204; Pride of Asia × C-2 and Pride of Asia × Red Cabbage; Pride of Asia × MR-1; 83-2 × Red Cabbage; and Pride of Asia × AC-204 and 83-2 × MR-1 were the best for Fe and Zn; Fe and Cu; Zn and Mn; Cu and Zn; and Cu, respectively. This study revealed clearly that none of the hybrids excelled for all the minerals suggesting the significance and need for multiple crossing breeding approaches, i.e. three way cross-hybrid, double cross-hybrid, population improvement, synthetics, composites, etc., for increasing the mineral concentration in cabbage head, i.e. “Breeding Cabbage for Higher Mineral” (Biofortification) without losing the vigour advantage for yield and other traits of economic importance to combat mineral deficiencies in human beings and plant systems.     

Factors affecting mango (Mangifera indica L.) protoplast isolation and culture

The major factors influencing protoplast isolation and culture of mango (Mangifera indica L.) cv. Kensington Pride were investigated. The resultant protocol was used to compare plating efficiency among 4 mango cultivars. Most responses differed between proembryonic masses (PEMs) and leaf sources. Protoplast yields of 15.22 × 10⁶ g⁻¹ from PEMs and 8.68 × 10⁶ g⁻¹ from greenhouse-derived leaves were obtained in a solution of 0.7 M mannitol CPW plus 1.5% cellulase, 1% hemicellulase and 0.75% macerozyme for PEMs or 0.5 M mannitol CPW plus 1.5% cellulose, 1% hemicellulase and 1.5% macerozyme for leaves. Culture in Ca-alginate beads with initial plating densities (IPD) of 2.5 × 10⁴ Pp mL⁻¹ for PEMs and 2.5 × 10⁵ for leaves gave the highest plating efficiencies (FPE). For PEMs 1 mg L⁻¹ 2,4-d and 3.5 mg L⁻¹ kinetin gave an FPE of 2.85% whereas lower kinetin (2 mg L⁻¹) plus 0.5 mg L⁻¹ 6-BAP was most effective for leaves (FPE of 2.12%). Most protoplast mortality occurred during the first week of culture and was more severe in liquid culture. In Ca-alginate beads, protoplast survival at 14 days was higher for PEMs (30%) than leaf (21%) as was the frequency of cell division (17.6% compared to 13.6%). PEMs protoplasts continued development through embryogenesis to in vitro plantlet regeneration whereas leaf protoplasts underwent cell division up to 40-cell colonies but failed to proceed further. For PEMs, polyembryonic cvs. Kensington Pride and Keow Savoey produced higher FPE (1.95%) than monoembryonic cvs. Tommy Atkins and Keitt (1.75%). There was no effect of cultivar for leaf protoplasts.     

Higher than optimum temperature under CO2 enrichment influences stomata anatomical characters in rose (Rosa hybrida)

There is little available information on the effects of temperature and CO₂ enrichment on stomata anatomical characteristics of plants. Effect of these two microclimates was studied on five rose (Rosa spp.) cultivars, viz. ‘First Red’ (used as check), ‘Arjun’, ‘Raktima’, ‘Raktagandha’ and ‘Pusa Pitamber’. Budded, single-stemmed rose cultivars having five lateral buds were grown in controlled environment growth cabinets under enriched CO₂ (1000 μmol mol⁻¹) and optimum (28/18 °C, T₀) or high (35/25 °C, T₁) temperature for 50 days. All observations were made on the abaxial leaf surface. Significant increases in stomatal density (68.7%), index (29.6%) and epidermal cell density (37.3%) were recorded in plants grown at high temperature over control with CO₂ enrichment. The cultivars responded differently in terms of length and width of guard cell and stoma (pore) under high temperature, however, the values averaged over treatments showed a significant reduction in these parameters. Further, number of stomata per leaf was higher (28.3%) in plants grown at high temperature, except First Red. A reduction in mean leaf area (26.7%) and dry mass (32.0%) was recorded at high rather than optimum temperature. The specific leaf area was maximum in Arjun (87%) while in First Red, a 14% reduction was noted at high temperature.     

Antioxidant enzymes in cabbage: Variability and inheritance of superoxide dismutase,peroxidase and catalase

Cruciferous vegetables are important source of dietary nutrients and antioxidants. Antioxidants have been touted as beneficial for enhancing plant stand and mitigating the effects of biotic and abiotic stresses. The objective of the present study was to determine variability for superoxide dismutase, peroxidase and catalase activity; its transmissibility; and correlation. Samples were harvested at fresh market stage, frozen immediately in liquid nitrogen and placed at −80 °C for assay. The genotypes, namely 83-2, Red Cabbage, KIRC-1-1, ARU Glory, Kinner Red, MR-1, AC-208, Red Rock Mammoth, KIRC-8 and KIRC-1A showed higher activity of antioxidant enzymes and therefore could be useful cascade in breeding for developing cultivars with high antioxidant activity. Enzymatic antioxidant activity showed 1.6, 12.8, and 18.2-fold difference for superoxide dismutase, peroxidase and catalase, respectively. Significant differences for enzymatic antioxidants among germplasm indicate towards the existence of substantial amount of variation. All enzymatic antioxidants showed high heritability along with low genetic advance as percentage of mean indicated the predominance of non-additive genes and provide good prospects for hybridization, and synthetics and hybrid development. The present information, therefore, could be used to develop antioxidant potential cultivars that will enhance in general the stress tolerance ability of plants, shelf life of produce, and healthy/better plant stand by enjoying antioxidants.     

H2O2 metabolism during sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ Xanthomonas axonopodis pv. citri interaction

Sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ is a canker (Xanthomonas axonopodis pv. citri: Xac) susceptible citrus genotype grown commercially worldwide. Canker causes severe economic losses and restricts the marketability of crop for export. Little is known about the role of oxidative stress in canker development. In the present investigation, sweet orange ‘Hamlin’ leaves were artificially inoculated with Xac to determine the impact of Xac infection on hydrogen peroxide (H₂O₂) metabolism. Characteristic symptoms following artificial inoculation were water soaking of the infiltrated zone between 2 and 8 days after inoculation (dai); raised epidermis accompanying tiny yellow colored bacterial colonies at 8 dai; and yellowing and necrosis of the infected zone by 12–16 dai. In planta Xac population increased 1000 fold by 14 dai from an initial population of 7.3 × 10⁶ cfu cm⁻² (0 dai). Peak concentrations of H₂O₂ were observed at 24 h and between 8 and 10 dai and coincided with higher activity of total superoxide dismutase (SOD). Lower levels of H₂O₂ in infected leaves were maintained by Xac induced higher activities of catalase (CAT), ascorbate peroxidase (APOD), and guaiacol peroxidase (POD). It appears Xac altered H₂O₂ metabolism in C. sinensis L. Osb. ‘Hamlin’ to enhance survival and growth.     

Anthocyanins from flowers of Hippeastrum cultivars

The anthocyanins, cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (1) and pelargonidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (2), were isolated from the ornamental flowers of a Ugandan Hippeastrum cultivar by a combination of chromatographic techniques, and their structures were elucidated mainly by the use of homo- and heteronuclear nuclear magnetic resonance spectroscopy and electrospray mass spectrometry. The same anthocyanins were found in six different Hippeastrum cultivars purchased in Norway. However, the absolute amount of the anthocyanins (0.08–1.79 mg/g, fresh weight) and the relative proportions of the individual anthocyanins varied from cultivar to cultivar (13.2–96.5% of 1). The colours of the fresh petals of the three cultivars ‘Red Lion’, ‘Royal Velvet’ and ‘Liberty’ were described by the CIELab coordinates L^* (lightness), C^* (chroma) and h_ab (hue angles). All the cultivars showed hue angles corresponding to scarlet nuances (h_ab = 22–35°), with the highest value in ‘Red Lion’. The most reddish petals (in ‘Royal Velvet’) contained the highest relative proportion of 1. Thus, the in vivo colours of these cultivars seem to be correlated with the relative proportions of individual anthocyanin in the petals.     

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.     

Growth and photosynthetic characteristics of blueberry (Vaccinium corymbosum cv. Bluecrop) under various shade levels

Blueberry can readily be shaded as a bush type plant, maybe affecting its growth and photosynthesis. Growth and photosynthetic characteristics of ‘Bluecrop’ blueberry grown under various shade levels were investigated to understand acclimation under shade conditions and to determine the optimal light conditions for agricultural purpose. Shade decreased the number of shoots per shrub, but increased shoot length. However, shade did not affect the number of leaves on the main axis. With increasing shade level, leaf length, width and area increased, but leaf thickness decreased. However, there was no obvious tendency in leaf length/width ratio with increasing shade level. Shade leaves had less dense stomata than sun leaves, but stoma was bigger in shade leaves than in sun leaves. With increasing shade level, non-photochemical quenching in blueberry leaves increased and the values were higher at low photosynthetic photon flux densities (PPFDs) in shade leaves than in sun leaves, resulting in the decreases in quantum yield, electron transport rate and net CO₂ assimilation rate (A_n). The maximum A_n at 31, 60, 73 and 83% shade levels was 11.8, 11.0, 8.4 and 7.5 μmol m⁻² s⁻¹, respectively. Following the slight decrease up to 100 μmol m⁻² s⁻¹ PPFD, stomatal conductance (g_s) linearly increased up to 600 μmol m⁻² s⁻¹ PPFD and became saturated at all shade levels. The leaves of the shrubs grown under the 83% shade level had a significantly lower g_s as compared to the leaves of the shrubs grown under the 31, 60 and 73% shade levels. Transpiration rate (E) linearly increased up to 600 μmol m⁻² s⁻¹ PPFD and was saturated at the 73 and 83% shade levels. However, E increased linearly at both 31 and 60% shade levels with increasing PPFD. The reproductive growth characteristics such as number of flowers, fruit set rate per flower bud and fruit yield also significantly decreased with increasing shade level. For agricultural purpose, therefore, shade level above approximately 60% of full sunlight must be avoided for optimal photosynthesis and growth of the ‘Bluecrop’ blueberry.     

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.     

Rapid seedlings regeneration from seeds and vegetative propagation with sucker and rhizome of Eremospatha macrocarpa (Mann & Wendl.) Wendl and Laccosperma secundiflorum (P. Beauv.) Kuntze

To develop efficient seedling production methods for Laccosperma secundiflorum and Eremospatha macrocarpa, a study was conducted to examine regeneration using offsets combined with several physical and chemical treatments of seeds. Offsets categorized into small, medium and large diameters, were planted in three conditions: shaded and open nursery, and greenhouse. We tested sucker from E. macrocarpa, and sucker and rhizome from L. secundiflorum. For both species, high viability percentage (ranging from 55% to 100%) were observed for small and medium suckers planted in shaded nursery and greenhouse, against less than 49% for sucker planted in open nursery. The mean seedling emergence times were estimated to 84, 77 and 75 days after planting (DAP) for small, medium and large sucker of L. secundiflorum, respectively under open nursery condition, and 76, 75, 95 DAP for small, medium and large suckers of the same species, respectively in shaded condition. Greenhouse has a significant positive effect on E. macrocarpa seedlings emergence time. For this species, the mean seedling emergence times were estimated to 43 DAP for small sucker and 76, 93 DAP for medium and large suckers. No seedling was obtained from rhizome planted in all the growing conditions tested. Concerning seed dormancy breaking, germination percentages and rates were determined for 13 treatments. The best treatments were pre-soaking unscarified seeds for 4 days in 1.01 g l⁻¹ and 0.10 g l⁻¹ KNO₃, with 79% and 68% of germination, respectively and in 3.46 × 10⁻³ g l⁻¹ GA₃ for 68% of germination. These methods are suggested to improve germination of L. secundiflorum seeds. Successful and recommended methods for E. macrocarpa are pre-soaking scarified seeds in 3.46 × 10⁻³ g l⁻¹ and 3.46 × 10⁻⁴ g l⁻¹ GA₃, 96% and 94% of germination, respectively. Dormancy, probably a combination of mechanical and chemical dormancy, is present in the two species.     

Hybridization and in vitro culture of an orchid hybrid Ascocenda ‘Kangla’

The present study focuses on hybridization program involving two species belonging to two different vandaceous genera, viz., Ascocentrum ampullaceum (Roxb.) Schltr. var. auranticum, a narrow endemic orchid of Manipur and Vanda coerulea Griff., an endangered orchid of Appendix I of CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora), to synthesize the primary hybrid genus with intermediate and improved characters in the F₁ generation. Observations on the crossability in the present bigeneric cross between V. coerulea and A. auranticum had been achieved with 60% success when V. coerulea was taken as female parent. Murashige and Skoog (MS) basal medium at half-strength was effective for the development of the hybrid seedlings of V. coerulea × A. auranticum followed by Vacin and Went (VW) and Knudson C (KC) media. The best response of seedling growth was observed on MS medium at half-strength supplemented with 2.3 μM kinetin + 0.5 μM α-naphthalene acetic acid with maximum shoot height (2.7 cm), leaf number (4.6) and root number (4.1) after 150 days of inoculation. The survival percentage and growth performance of the seedlings were found to be higher (80% survival) in potting substrate consisting of brick:charcoal in the ratio 2:1 mulched with moss (Sphagnum sp.) than in potting substrate consisting of brick:charcoal:tree fern in the ratio 2:1:1. The first flowering was observed in the hybrid seedlings of V. coerulea × A. auranticum after 2 years of transfer to the ex vitro environment. Morphologically, the flowers differed from that of the parents clearly showing the success of the hybridization experiment. Registration of the hybrid has been made with the Royal Horticultural Society with the nomenclature Ascocenda ‘Kangla’ (No. T128725).     

Analysis and modeling of gas exchange processes in Scaevola aemula

Scaevola aemula is a popular ornamental crop cultivated as a bedding plant or for hanging baskets. We characterized gas exchange properties of S. aemula ‘New Wonder’ in response to photosynthetically active radiation (PAR), carbon dioxide concentration, and leaf temperature. Net CO₂ assimilation rate (A) was responsive to CO₂, exhibiting a saturation when intercellular CO₂ concentration (C_i) was greater than 600 μmol mol⁻¹. Net CO₂ assimilation rate and dark respiration rate (R_d) were 23.1 and 2.3 μmol m⁻² s⁻¹, respectively, at 25 °C and PAR = 1500 μmol m⁻² s⁻¹. Net CO₂ assimilation rates were similar at leaf temperatures between 20 and 30 °C but significantly reduced at 15 °C. These gas exchange results were used to test the extendibility of a coupled gas exchange model previously developed for cut-roses. Utilizing the gas exchange data measured at 25 °C leaf temperature, several model parameters were independently determined for S. aemula. Model predictions were then compared with observations at different leaf temperatures. The model predicted the rates of net CO₂ assimilation and transpiration of S. aemula reasonably well. Without additional calibration, the model was capable of predicting the temperature dependence of net CO₂ assimilation and transpiration rates. Applying the model to predict the effects of supplemental lighting and CO₂ enrichment on canopy photosynthesis and transpiration rates, we show that this model could be a useful tool for examining environmental control options for S. aemula production in the greenhouse.     

Aluminium-induced effects on growth,morphogenesis and oxidative stress reactions in in vitro cultures of quince

The effects of Al³⁺ [supplied as Al₂(SO₄)₃·18H₂O] addition to culture media (pH 4.0) on growth, morphogenesis (in leaf explants), and oxidative stress reactions in in vitro cultures of ‘BA 29’ quince were investigated. Aluminium (Al 0.5 mM) strongly inhibited shoot growth in the proliferation and rooting phases (Al 2.2 mM), reduced shoot proliferation (Al 1.1 mM), and induced tissue browning. Superoxide dismutase (SOD) activity was increased in shoot cultures supplemented with 2 mM Al. Malondialdehyde (MDA) content of shoots was strongly increased by Al during proliferation (starting from Al 1.7 mM) and rooting (already at Al 1.1 mM), thus serving as a good ‘marker’ for Al toxicity. Even a low concentration of Al (0.5 mM) in the shoot induction medium was found to inhibit shoot regeneration. When standard (Al 0) shoot induction medium was used, leaf explant growth was only reduced by 2.2 mM Al in the subsequent growth phases. Following a preliminary selection for their growth on Al-enriched media, 82 potentially Al-tolerant quince somaclones were selected for further trials.     

Production protocol development for greenhouse cut Linaria, Lupinus, and Papaver flowers

Linaria maroccana Hook. f. Ann., ‘Lace Violet’, Lupinus hartwegii ssp. cruikshankii Lindl. ‘Sunrise’ and Papaver nudicaule L. ‘Meadow Pastels’ seeds were directly sown into 105 cell plug trays and received either ambient light or supplemental high intensity discharge (HID) lighting. For each species, a 2 × 3 × 3 factorial was used with two light intensities during propagation, three transplant stages, and three night temperatures. Seedlings were transplanted at the appearance of 2–3, 5–6, or 8–9 true leaves. Transplanted Linaria and Papaver seedlings were placed at 5/11, 10/16, or 15/21 ± 1 °C night/day temperatures and Lupinus seedlings were placed at 15/24, 18/25, or 20/26 ± 2 °C night/day temperatures. For this study, the optimum production temperature for Linaria was 10/16 °C as the cut stems produced at 15/21 °C were unmarketable and production time was excessively long at 5/11 °C. At 10/16 °C, Linaria seedlings should be transplanted at the 2–3 leaf stage to maximize stem number, stem length and profitability. For Lupinus the optimum temperature was 15/24 °C due to long stems and high profitability per plant. Lupinus seedlings should be transplanted at the 2–3 leaf stage when grown at 15/24 °C to obtain the longest and thickest stems; however, $/m² week was higher for plants transplanted at the 8–9 leaf stage due to less time in finishing production space. For Papaver, the 15/21 °C temperature was optimal as that temperature produced the longest stems in the shortest duration, resulting in the highest $/m² week. At 15/21 °C Papaver plants should be transplanted at the 2–3 leaf stage. Supplemental HID lighting had no effect on any of the species.     

Production of inter-section hybrids between Primula filchnerae and P. sinensis through ovule culture

Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l⁻¹ sucrose, 2.5 g l⁻¹ gellan gum, 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 0.1 mg l⁻¹ 6-benzyladenine (BA) and 50 mg l⁻¹ gibberellic acid (GA₃). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l⁻¹ sucrose and 3 g l⁻¹ gellan gum, without plant growth regulators (PGRs) or with 1 mg l⁻¹ zeatin and 0.1 mg l⁻¹ NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.     

Interspecific hybridization in Lonicera caerulea and Lonicera gracilipes: The occurrence of green/albino plants by reciprocal crossing

Lonicera caerulea L. var. emphyllocalyx (Maxim.) Nakai is a berry crop cultivated in cold regions. So far, commercial cultivars have been mainly introduced from selection of wild plants. Therefore, fruit traits and other agricultural characteristics have been limited. In this study, interspecific crosses between L. caerulea var. emphyllocalyx and Lonicera gracilipes var. glabra Miquel were examined to increase genetic variability of L. caerulea var. emphyllocalyx. Seedlings were obtained from reciprocal crosses between L. caerulea var. emphyllocalyx and L. gracilipes var. glabra. The hybrid nature of seedlings was confirmed with random amplified polymorphic DNA analysis. Viable green plants were obtained efficiently from L. gracilipes var. glabra × L. caerulea var. emphyllocalyx. In contrast, all plants produced from L. caerulea var. emphyllocalyx × L. gracilipes var. glabra were albino. These albino plants were very weak and only survived in culture condition. The chlorophyll deficiency was unilaterally observed, suggesting the occurrence of nuclear–cytoplasmic incompatibility. Viable F₁ hybrids obtained from L. gracilipes var. glabra × L. caerulea var. emphyllocalyx are amphidiploid (2n = 4x = 36) as showing same to both parents. The hybrid plants are expected to increase the variability of fruit traits, and may have heat tolerance from L. gracilipes var. glabra.     

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.     

Cold storage of oil rose (Rosa damascena Mill.) flowers

Oil rose flowers were stored at 0 °C in four different packaging materials [plastic box + stretch film (PB + SF), Xtend^®, Smartbag^® and polyethylene (PE)] for 60 days. During storage, weight loss, O₂ and CO₂ concentrations in the packages, petal color and sensorial attributes were investigated besides essential oil content and composition. Storage duration and packages had significant (p < 0.01) effects on weight loss. At the end of storage, the lowest weight loss was in PE package (1.696%) whereas the highest weight loss was in Xtend^® (10.081%). The essential oil content was significantly (p < 0.01) affected by storage duration and packages. In addition, the essential oil contents obtained from all packages for a storage period of 10 days and the essential oil contents obtained from unstored (control) petals were included in the same group. At the end of storage, the essential oil contents decreased by 91.3, 57.7, 80.0 and 64.3% in PB + SF, Xtend^®, Smartbag^® and PE packages, respectively as compared to control. In addition, storage duration and package types significantly (p < 0.01) affected petal color, O₂ and CO₂ concentrations in the packages and sensorial scores. The concentration of citronellol, a main component of rose oil, increased in all packages during storage of 10 days in comparison to the control group while it varied in other storage durations and package types. However, nerol and geraniol were lower than the control group during storage while concentrations of nonadecane, heneicosane and eicosane were higher. In conclusion, loss of oil yield and quality, due to various reasons and particularly due to fermentation in oil rose from the harvest of petals to their distillation, can be minimized with storage of petals in all package types for up to 10 days.     

Conservation of Vanilla species, in vitro

Vanilla (Vanilla planifolia) is a crop of great commercial importance as the source of natural vanillin, a major component of flavor industry. The primary gene pool of V. planifolia is narrow and is evidently threatened due to destruction of its natural habitats making the secondary gene pool important as a source of desirable traits especially for resistance to diseases. Many species of vanilla are considered rare and endangered hence an urgent need to conserve them, arises. Effective procedures for micropropagation and in vitro conservation by slow growth in selected species of vanilla, are described. Synthetic seed technology was standardized by encapsulating 3–5 mm in vitro regenerated shoot buds and protocorms in 4% sodium alginate, which could be stored up to 10 months with 80% germination in sterile water at 22 ± 2 °C. In vitro conservation technology of Vanilla was standardized and shoot cultures could be maintained for more than 1 year without subculture, on slow growth medium, i.e. Murashige and Skoog medium supplemented with 15 g l⁻¹ each of sucrose and mannitol in sealed culture vessels at 22 ± 2 °C. These cultures were maintained in vitro for more than 7 years with yearly subculture. The conserved material could be retrieved and multiplied normally in MS medium with 1.0 mg l⁻¹ BA and 0.5 mgl ⁻¹ IBA. The in vitro conserved plants showed good growth and developed into normal plants. This synseed and in vitro conservation system can be utilized for conservation and exchange of vanilla genetic resources.     

Gibberellin producing Neosartorya sp. CC8 reprograms Chinese cabbage to higher growth

Gibberellins production by soil fungi received little attention, although substantial work has been carried out on other growth promoting aspects of soil borne fungi. We investigated gibberellins production and growth promoting capacity of a novel fungal strain of Neosartorya, which was isolated from the roots of Chinese cabbage (Bassica rapa L. ssp. pekinensis). Fungal culture filtrates (CF) obtained from pure cultures of 16 endophytic fungi were bioassayed on Waito-C, in order to investigate plant growth promoting capacity of these fungi. The fungal isolate CC-8 induced maximum shoot length of Waito-C (13.0 cm) as compared to control treatments. In a separate experiment, the CF of fungus CC-8 significantly promoted plant length and biomass of Chinese cabbage. The fungal CF also increased endogenous bioactive GA₁ and GA₄ contents of Chinese cabbage. Gibberellin analysis of CF of CC-8 showed presence of both physiologically active and non active gibberellins in higher concentrations (GA₁, 1.42 ng/ml; GA₃, 5.93 ng/ml; GA₄, 11.36 ng/ml; GA₇, 3.25 ng/ml; GA₉, 0.79 ng/ml; GA₁₅, 1.18 ng/ml). The culture filtrate of CC-8 produced higher amounts of GA₃, GA₄, GA₇ and GA₉ than wild type Fusarium fujikuroi, a well known gibberellins producing fungus. The fungal isolate CC-8 was later identified as a new strain of Neosartorya species on the basis of traditional and advance molecular techniques.