Callus induction and culture of Rosa

Leaf and stem explants of Rosa manetti Hort. and R. hybrida L. ‘Tropicana’ were placed on Murashige and Skoog (MS), Schenk and Hildebrandt (SH) and Gamborg and Wetter (1-B5C) media, containing growth regulators, casein hydrolysate (CH) or coconut milk (CM), to determine the optimum conditions for callus initiation and maintenance. The explants were cultured either in dark or in light (2 Klux 16 h/day) at 26±2°C. Friable, fast growing callus was evident after 3 weeks for both rose species on MS + 2.0 mg/l 2,4-D, 0.25 mg/l kinetin (K) and 2.0 g/l CH as well as on SH + 0.5 mg/l 2,4-D, 2.0 mg/l p-CPA and 0.1 mg/l K. Callus initiation occurred sooner on SH than on MS. However, during sub-culture of callus arising on SH, rapid oxidation often occurred, resulting in severe browning of the callus, which made it unsatisfactory for further experimental use. Callus initiation occurred faster in dark than in light, but deteriorated when continuously sub-cultured in the dark regardless of media. Although ‘Tropicana’ initiated callus sooner than R. manetti, very fast growing callus of the latter was obtained when leaf callus was transferred to MS + 2.0 mg/l NAA and 0.5 mg/l BA. Unsuccessful attempts to regenerate callus and induce adventitious shoots on the 2 explants are discussed.     

Guazatine control of sour rot in lemons,oranges and tangors under various storage conditions

Guazatine (1-17-diguanidino-9-aza-heptadecane acetate) was compared with sodium orthophenylphenate (SOPP), sec-butylamine (2-AB), benomyl and tridemorph for the control of citrus sour rot (Geotrichum candidum) under local marketing, export, and ethylene (C₂H₄) degreening storage conditions. Guazatine at 25–125 mg/l gave better control than 20 g/l SOPP, 10 g/l 2-AB and 250 mg/l benomyl in artificially inoculated ‘Eureka’ lemons stored at 27° C and high humidity for 6 days. At 500 mg/l, guazatine gave better control than 20 g/l SOPP, 10 g/l 2-AB or 500 mg/l benomyl in dip-inoculated ‘Eureka’ lemons, treated and packaged as for export and stored at 7° C for 2 or 10 weeks. At 250 mg/l, guazatine provided better control than 20 g/l SOPP, 5 g/l 2-AB, 250 mg/l benomyl or 1000 mg/l tridemorph in artificially inoculated ‘Washington’ navel oranges held for 4 days under degreening-conditions of 27° C and 90% RH with added C₂H₄.     

Rapid propagation of white cabbage by tissue culture

A tissue culture technique has been devised to produce plants of white cabbage (Brassica oleracea v. capitata L.f. alba) from heads stored at 0.5 ± 0.5°C for 8 months. Meristem-tips (0.5–2 mm diameter), excised from heads of 11 accessions, were grown initially on MS medium containing 2.56 mg l^?1 of kinetin and then induced to proliferate shoots on MS medium with 12.8 mg l^?1 kinetin. Subsequent transfer to a kinetin-free medium resulted in root development in 1–2 weeks. Rooted plantlets were readily established in soil. Plantlets obtained in this way from parent cabbages containing turnip mosaic virus remained infected.     

Effect of gibberellic acid and Vapor Gard on ripening,amylase and peroxidase activities and quality of mango fruits during storage

Post harvest application of gibberellic acid at 200 mg 1^?1, Vapor Gard (di-l-p-menthene) at 2.5% and their combination was studied on ‘Mallika’ mangoes (Mangifera indica L.) stored at ambient temperature (37 ± 2° maximum and 34 ± 2°C minimum) and at 15°C. Significant delay in the ripening of mango fruits was observed when gibberellic acid was applied with or without Vapor Gard. Gibberellic acid significantly retarded the degradation of ascorbic acid and chlorophyll in the peel, and reduced a-amylase and peroxidase activities during storage. Loss of weight decreased following treatment with Vapor Gard either alone or with gibberellic acid during storage at both ambient temperature and at 15°C. A pronounced retardation of ripening was observed when fruits were treated with gibberellic acid and Vapor Gard and stored at 15°C. The study thus suggests that mango fruits can be successfully stored for 20 d by application of gibberellic acid (200 mg 1^?1) in combination with Vapor Gard (2.5%) and stored at 15°C.     

Effect of stratification-temperature,chemical treatments and seed coat on the growth of peach seedlings cultivar ‘Sharbati’

In order to obtain normal seedlings of peach cultivar ‘Sharbati’ before the commencement of winter, treatments with GA₃, thiourea and kinetin were given to seeds before stratification at 7°C, 10°C or 24°C. The seedlings raised from the treated seeds and after-ripened at 24°C were dwarf. The seedling growth was increased when the treated seeds were stratified at 10°C or 7°C and the stratification period was prolonged from 15 days to 75 days. 10°C stratification-temperature was better than 7°C. The seedling growth was improved when the seed coat was removed before the treatments. With respect to both seed types, 1000 mg/l GA₃ produced the tallest seedlings at all the after-ripening temperatures and during each stratification period. The next best treatment was 100 mg/l kinetin.     

In vitro propagation of Lychnis senno Siebold et Zucc., a rare plant with potential ornamental value

Lychnis senno is a rare and valued ornamental plant. Seed propagation is not efficient because of the low germination rate. To grow commercially L. senno in China, a protocol for in vitro germination and propagation of this species was developed. Various germination rates were obtained by treating seeds with GA₃ during 1–6 months storage period. The highest germination rate reached 19.4% when seeds were treated with 250 mg/l GA₃ and stored for 5 months at 4 °C. Axillary shoot proliferation was induced in the nodal segments of the seedlings on medium containing specific concentrations of BA and NAA [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant 15, 473–497]. Maximum number of shoots was developed on a medium supplemented with 5 mg/l BA and 0.5 mg/l NAA, while the higher shoots were observed on a medium supplemented with 0.5 mg/l BA and 0.05 mg/l NAA. Rooting was induced in 91.7% of the regenerated explants on a half-strength MS medium supplemented with 0.5 mg/l NAA. The plantlets grew well and flowered after transfer to the greenhouse. The chromosome numbers of seedlings and propagated plants were also determined to be 2n = 2x = 24.     

Conservation of Zingiber germplasm through in vitro rhizome formation

The effects of various concentrations of maleic hydrazide (MH; 2, 4, 6, 8 mg/l) and three light treatments (16-h, 24-h, 0-h) on in vitro rhizome formation and conservation of ginger (Zingiber officinale Rosc. cv. Rio de Janeiro) were studied. In vitro rhizome formation occurred in all the above treatments. Addition of MH (2–8 mg/l) to the control medium (CM) comprising Murashige and Skoog's (1962) salts, 9% sucrose, 0.8% agar-agar, 0.1 mg/l α-naphthaleneacetic acid (NAA), 1 mg/l N⁶-benzyladenine (BA), did not show any significant positive effects on rhizome formation as well as survival of cultures. A significant effect of light treatments was observed on survival of cultures but not on rhizome formation. More than 50% cultures survived up to 14 months on CM under 16-h and 24-h light conditions as compared to 20% cultures on same medium incubated under dark. A total of 33 genotypes of cultivated and wild species of Zingiber were subsequently tested for conservation through in vitro rhizome formation on CM under 16-h light condition. All genotypes produced rhizomes of varying size with numbers ranging from 3 to 15 per culture and were conserved for at least 12 months; some genotypes could be conserved even up to 16–20 months. Viability of rhizomes was determined by in vitro regeneration of shoots upon subculture and their subsequent establishment in soil. Following the protocol described in the present paper, some 160 genotypes of cultivated and wild species of Zingiber, collected from different geographical regions of India, are being conserved at In Vitro Genebank of National Bureau of Plant Genetic Resources, New Delhi.     

Response of the chrysanthemum to soil heating

Chrysanthemum plants were exposed to 16°C day-temperature, 11°C night-temperature, 13°C soil-temperature, to be indicated as 16/11/13°C, or to 16/11/25°C, 20/16/18°C, or 20/16/25°C, first long day, then short day, (long day = 12-h light period with 3-h night break; short day = 12-h light period) from planting to harvest in controlled environments to study the effects of soil heating on growth and flowering. There were significant, but not substantial, effects of soil heating on leaf area, percent soluble carbohydrate, flower bud diameter, days to visible bud and some other parameters. Two winter cultivars responded similarly, while 2 summer cultivars differed in flowering-response to soil heating. An experiment was also conducted using 16/11/25°C day/night/soil temperatures during long days, short days or throughout the complete growth cycle, with 16/11/13°C day/night/soil temperatures at other times. Soil heating during long days resulted in the highest quality flowers. Soil heating during short days or throughout the growth period resulted in most rapid flowering but decreased flower quality.     

Phenol induced browning and establishment of shoot-tip expiants of 'Fuji' apple and 'Jinhua' pear cultured in vitro

SUMMARY

Several factors influencing phenol induced browning and establishment of apple (Malus pumila Mill. cv. Fuji) and pear (Pyrus bretschneideri Rend., cv. Jinhua) were investigated by using shoot-tip expiants cultured in vitro. Expiants of 'Fuji' apple and 'Jinhua' pear collected from November to February produced low browning percentages. Browning percentages increased rapidly as the stock plants entered the growing season, reached a maximum during April and August and then decreased. Dark treatment of the stock plants reduced browning. Lowest incidence of browning and the highest surviving percentages of both 'Fuji' apple and 'Jinhua' pear were obtained after four weeks of dark treatment. However, bud-break percentage, fresh weight and the number of new leaves of the surviving expiants decreased with increase in the length of dark treatment. When the dark treatment was used during the first phase of culture initiation, 5°C was better than 24°C for reducing browning and improving establishment of the expiants. At 5°C, 6 d of dark treatment for 'Fuji' apple and 8 d for 'Jinhua' pear produced the least browning and the highest surviving percentages. However, bud break percentages, fresh weights and numbers of new leaves decreased as the length of the dark treatment increased. Addition of 2.5 g l"1 activated charcoal to the initiation medium favoured establishment of 'Jinhua' pear expiants and 100/150 mg 1"' ascorbic/citric acid were effective with 'Fuji' apple.     

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.     

In vitro Germination of Early Ripening Sweet Cherry Varieties (Prunus avium L.) at Different Fruit Ripening Stages

In vitro embryo culture enabled satisfactory germination of immature seeds produced in crosses from early ripening sweet cherry varieties (Prunus avium L.). Three varieties —‘Rita’, ‘Bigarreau Burlat’ and ‘Carmen’— were crossed with ‘Early Star’ as male parent. Germination rate was affected by the developmental stage of both fruit and embryo. Fruit ripening stage was a critical factor for culture infection rate that increased with maturity. In-ovule embryo culture on Murashige and Skoog medium without hormones improved the embryo size but did not increase the germination rate due to a further increase in infection rate. Ex-ovule embryo culture on Murashige and Skoog medium supplemented with BA 1 mg L^?1, NAA 0.5 mg L^?1, 20 g L^?1sucrose, 10 g L^?1 sorbitol and 6 g L^?1agar during the stratification time increased embryo length. Germination was performed on Brooks and Hough medium at the 22?±?1?°C with 16/8 h light/dark photoperiod. The highest germination rate (75?%) was reached in embryos that were 3?4 mm in length, after 30-days stratification at 4?°C. Embryos in fruits at green-yellow stage that were 3?4 mm long were morpho-physiologically developed to produce bipolar seedlings, without combined application of embryo culture and micropropagation.     

In vitro production of Prunus necrotic ringspot virus-free begonias through chemo- and thermotherapy

Prunus necrotic ringspot virus (PNRSV)-free Begonia spp. plants were raised from petioles of virus-infected plants using in vitro techniques. The petioles were grown on MS medium supplemented with 0.2 mg/l NAA and 0.2 mg/l BAP (pH 5.8). For rooting, half-strength MS medium without any plant growth regulators was used. On rooting medium, shoots were subjected to chemotherapy (virazole, 2-thiouracil or 6-azauracil) and thermotherapy (38 °C for 16 h light period and 22 °C for 8 h dark period) separately or in combination. Regenerated plants (treated with chemo- and thermotherapy) were indexed for PNRSV by DAS-ELISA and RT-PCR. An amplified product of 785 bp was obtained by RT-PCR in PNRSV-infected plants. Virazole at a concentration of 20 mg/l was found to be more effective (30 and 20% of PNRSV-free plants as indexed by ELISA and RT-PCR, respectively) in comparison to the other chemicals. Thermotherapy for 25 days gave 35 and 25% PNRSV-free plants as indexed by DAS-ELISA and RT-PCR, respectively. A combination of both treatments gave a good number of PNRSV-free plants (67.5 and 57.5% as indexed by DAS-ELISA and RT-PCR, respectively). At higher concentrations all three chemicals were found to be toxic. Thermotherapy for more than 25 days caused browning of leaves and shoots died.     

Plantlet regeneration and stability from callus cultures of Freesia × hybrida Bailey cultivar ‘Royal’

True-to-type plantlets of Freesia × hybrida Bailey cultivar ‘Royal’ were generated from callus after 27 months of sub-culture in liquid medium. Callus was initiated from young flower pedicels cultured on semi-solid Linsmaier—Skoog (LS) medium supplemented with 5 mg/l of 2,4-D and 0.5 mg/l kinetin, and transferred to the same medium in liquid form without hormones and thereafter sub-cultured every 7–10 days. Liquid cultures with 2.4–4.3 g of callus per 25 ml medium produced largest increases in callus fresh weight. Callus generated the most shoots when cultured on LS medium supplemented with 5 mg/l of kinetin and incubated in the light, while fewer plantlets were produced when no growth regulator or GA₃ or PBA were used. Callus cultures incubated in continuous darkness did not form shoots.     

Response of kiwifruit (Actinidia deliciosa cv. Allison) to post-harvest treatment with 1-methylcyclopropene

Summary

Experiments were conducted to observe the effect of different concentrations of 1-methylcyclopropene (1-MCP) on the post-harvest life and quality of ‘Allison’ kiwifruit (Actinidia deliciosa). Fruit were treated with 1-MCP at 0.5 µl l^–1, 1.0 µl l^–1, or 2.0 µl l^–1, un-treated fruit served as controls. Each 1-MCP treatment was applied for 24 h at 20°C. After treatment, fruit were transferred to ambient temperature storage (22º ± 4ºC; 65 – 70% relative humidity) for 18 d, during which time observations on various physical, physiological, and biochemical parameters were recorded at 3 d intervals. Our results indicated that 2.0 µl l^–1 1-MCP was the most effective treatment to delay softening and ripening in ‘Allison’ kiwifruit, as such fruit showed the lowest mean weight loss (9.8 ± 0.2%), the highest mean fruit firmness value (32.7 ± 0.2 N), and began to ripen only after 12 d in storage, whereas untreated fruit started ripening on day-6 of storage. The activities of fruit softening enzymes such as polygalacturonase (PG; 58.5 ± 0.3 µg galacturonic acid g^–1 FW h^–1), and lipoxygenase (LOX; 3.96 ± 1.3 µmoles linoleic acid oxidised min^–1 g^–1 FW h^–1) were lower, and total phenolics (TP) contents (24.3 ± 0.3 mg 100 g^–1) and anti-oxidant (AOX) activities (12.5 ± 0.03 µmol Trolox g^–1 FW h^–1) were higher in 1-MCP-treated fruit than in untreated fruit (PG, 98.3 ± 0.5 µg galacturonic acid g^–1 FW h^–1; LOX, 4.39 ± 1.0 µmoles min^–1 g^–1 FW h^–1; TP, 5.3 ± 0.6 mg 100 g^–1; AOX, 4.7 ± 0.02 µmol Trolox g^–1 FW h^–1, respectively). In addition, 1-MCP-treated fruit exhibited lower rates of respiration (48.3 ± 0.4 ml CO₂ kg^–1 h^–1) and ethylene production (30.2 ± 0.02 µl kg^–1 FW h^–1) than untreated fruit (58.9 ± 0.6 ml CO₂ kg^–1 h^–1; 38.7 ± 0.04 µl kg^–1 FW h^–1, respectively). Similarly, 1-MCP-treated fruit had higher titratable acidity (TA; 1.33 ± 0.3%) and ascorbic acid (AA) contents (115.9 ± 2.6 mg 100 g^–1 pulp) and lower soluble solids contents (SSC; 8.33º ± 0.2º Brix) than untreated kiwifruit (TA, 1.0 ± 0.2 %; AA, 105.3 ± 2.2 mg 100 g^–1 pulp; SSC, 13.7º ± 0.3º Brix, respectively). Thus, 2.0 µl l^–1 1-MCP can be used for the post-harvest treatment of ‘Allison’ kiwifruit to enhance its shelf-life and marketability by approx. 6 d.     

Rooting of peach hardwood cuttings for the meadow orchard

In order to make peach meadow orchard systems feasible, the possibility of propagating peaches by hardwood cuttings and thus reducing the establishment cost of this extremely high-density planting system was examined.A medium temperature of 25°C was harmful to auxin-treated peach cuttings, but excellent rooting could be obtained at a temperature as low as 12°C. Further trials conducted under outdoor conditions in winter in the coastal plain of Israel, with a soil temperature of 12–14°C, did not show any effect of timing on rooting, providing the cutting was taken mature enough (after mid-November) and not later than about 6 weeks prior to bud break. In 30 days root initiation started. Although variations were found between cultivars, 1500 mg/l of indole butyric acid in a 5-second base-dip was found to act well with most cultivars. Good aeration was shown to be critical for good root formation. Leaving the cutting for 1 month in damp sand prior to planting in orchard soil was satisfactory. An area of a few hectares of commercial meadow orchards has already been established with this method.     

In vitro morphogenesis and growth of Narcissus in response to temperature

In vitro morphogenesis and growth of tissues of the Narcissus cultivar ‘Lord Nelson’ have been assessed in constant and alternating temperatures. Tissue of leaf-base and scape origin, producing leafy shoots and bulblets on a defined sterile medium, were grown at constant and alternating temperatures of 15, 20, 25 or 30°C and 15–25 or 20–25°C, respectively, in a 16-h daily photoperiod. The number of leaves, leaf length, volume of tissues, fresh weight (but not percentage dry matter), and the number of shoots that would ultimately produce bulbs, were maximal at a constant temperature of 25°C as compared with all other temperature-regimes tested.     

Increases in ethylene and carbon dioxide production by Tulipa gesneriana L. ‘Prominence’ after completion of the cold-requirement

Bulbs of Tulipa gesneriana L. ‘Prominence’ were either specially pre-cooled at 5 ± 0.5°C or held at 17 ± 0.5°C in a flow-through system equipped for atmospheric sampling. Bulbs at 17°C had low CO₂ and C₂H₄ production rates until January when they began to increase. An initial peak of C₂H₄ production occurred during the 2nd week of pre-cooling, followed by a major increase after 12 weeks. In addition, bulbs were specially pre-cooled for periods of 2–16 weeks (2-week increments). The bulbs were then transferred to 17 ± 0.5°C, where initial periods of special pre-cooling of greater than 12 weeks resulted in a dramatic increase in respiration rate over bulbs cooled for less than 12 weeks. These increases in C₂H₄ and CO₂ liberation appeared to be related to completion of the bulb cold-requirement. However, no surge of shoot elongation occurred after 12 weeks of pre-cooling and transfer to 17°C.     

Silver nitrate and adenine sulphate induced high regeneration frequency in the recalcitrant plant Cosmos bipinnatus using cotyledon explants

Plant regeneration ability was studied in the medicinal-ornamental plant, Cosmos bipinnatus ‘Sonata white’, which is a dicotyledonous recalcitrant plant to shoot induction. Cotyledons were used as sources of explants to investigate plant regeneration. High frequency of direct shoot induction was obtained when BA (5 mg/l) and AgNO₃ (5 mg/l) were used in combination with 20 mg/l adenine sulphate (73.8%) in Murashige and Skoog (MS) medium. The highest shoot number per explant (5.7) was induced on MS medium supplemented with 5 mg/l BA, 5 mg/l AgNO₃, and 40 mg/l adenine. Eight week-old shoots were transferred to root induction media containing MS and half-strength MS medium with different concentration of IBA. The highest rate of root induction (70.8%) was obtained on half-strength MS medium with 1.5 mg/l IBA within four weeks. The plantlets were transferred to pot and kept in the greenhouse condition. Seventy percent of the plantlets successfully acclimatised.

Abbreviations: BA, 6-benzylaminopurine; IBA, Indole-3-butyric acid; MS, Murashige and Skoog; PGRs, plant growth regulators.     

Low temperature and gibberellic acid stimulation of germination in Sambucus caerulea Raf

Sambucus caerulea (elder) seeds did not germinate after 4°C cool treatments for up to 30 days, when monitored for a further 30 days at 21°C. When seeds were soaked for 24 h in gibberellic acid (GA₃) prior to and during cold treatment, germination percentage depended on GA₃ concentration and duration of cold treatment. The highest germination percentage was 55 (1000 mg l⁻¹ GA₃ for 30 days at 4°C). When seeds were treated with ethephon at 0, 100 or 1000 mg l⁻¹, no germination was recorded after a subsequent 30-day 4°C treatment. Ethephon added to GA₃ gave a strong interaction, leading to further promotion in germination. Optimal germination was obtained after 1000 mg l⁻¹ GA₃ and 100 mg l⁻¹ ethephon for 30 days at 4°C (69%).The addition of ethanol, acetone, dimethyl sulfoxide or polyethylene glycol to the GA₃ soak as infusion agents either reduced or did not change the germination percentage.     

Effects of low oxygen short-term exposure at 15°C on postharvest physiology and quality of apricots harvested at two ripening stages

Summary

Apricots of two harvests (9–10° Brix and 13–14° Brix) were treated for 6 d at 15°C with 1%, 2% and 4% O₂ (low oxygen = LO) and then kept for 7 d and 5 d (respectively for the first and the second harvest) in air at 15°C (shelf life = s.l.). Control fruit were held continuously in air at 15°C or for 6 d at 5°C then moved to 15°C. In early harvest fruit, low O₂ (1% and 2%) for 6 d controlled ethylene production even after the transfer and consequently the fruit had lower SCC and were firmer. In fruit from the second harvest, only 1% O₂ atmosphere or control fruits at 5°C were able to control the rise of ethylene in s.l. and to reduce the increase of soluble solids content (SSC). A 1% O₂ atmosphere maintained acceptable firmness even during s.l. in fruit of both harvests. Respiration rate was better restricted by low temperature during the treatment but at the end of experiment no difference was observed among the samples. Colour of apricots was maintained only by 1% O₂ atmosphere or 5°C temperature both in the first and in the second harvest. Sensory evaluation of fruit of the first harvest revealed that only apricots kept at 15°C or in 4% O₂ were considered saleable. In the second harvest, apricots treated with 5°C and 1% O₂ were judged saleable. In conclusion, early harvest fruit does not benefit from low oxygen (1% and 2% O₂) because fruit does not reach the optimal SSC whereas for late harvest apricots the use of 1% O₂ at a higher temperature than that used commercially can be an alternative to low temperature as shipping treatment or short term storage.