Tissue culture propagation of Lilium hybrids

We studied the effect of growth regulators-α-naphthyl-acetic acid (NAA) and 6-benzylaminopurine (BAP) on bulblet regeneration from bulb scale explants of Lilium oriental hybrid ‘Crimson Beauty’. The combination of 5 μM BAP and 1 μM NAA in the basal culture medium of Linsmaier and Skoog induced maximum bulblet formation per explant. On the same medium we succeeded in inducing bud differentiation on the ovary- and leaf-derived explants and in callus culture. A method is described for a continuous propagation of lily plants in vitro.     

An efficient adventitious shoot regeneration system on excised leaves of micropropagated lingonberry (Vaccinium vitis-idaea L.)

Summary

An efficient system to regenerate shoots in vitro on excised leaves of lingonberry (Vaccinium vitis-idaea L.) was developed. Leaf explants from shoot-proliferating cultures produced multiple shoots without an intermediary callus phase on zeatin (ZN)-containing shoot induction media within 3–4 weeks of culture initiation. Cultivars Regal and Splendor, and one clone from a natural stand in Estonia (ECL1), were used in the first experiment. Young expanding leaves with the adaxial side touching the culture medium, and maintained for 7 d in darkness, produced the best results. There were significant genotypic differences in adventitious shoot formation. A second experiment studied the effects of ten concentrations of three cytokinins: ZN at 5, 10, 20, 30 and 40 μM; 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron, TDZ) at 0.1, 1, 5 and 10 μm; and 6-(γ-γ-dimethylallylamino) purine (2ip) at 25 μM were compared with leaf segments of different polarity in ‘ECL1’. Zeatin was found to be more effective than TDZ or 2iP as an inductive signal for regenerating many vigorous shoots. Zeatin induced multiple shoot formation at all concentrations tested, but maximum morphogenic response was observed at 20 to 30 μM. The media containing TDZ generally promoted more callus formation and suppresed shoot elongation. In a third experiment with the lingonberry cultivar Erntedank and the clone ‘ECL1’, a new medium developed for lingonberry shoot culture proved more effective than the modified Murashige and Skoog medium for regenerating shoots on leaf explants. Elongated shoots were excised and rooted directly on a 2 peat:1 perlite (v/v) medium after dipping in 0.8% indole-3-butyric acid. Rooted plantlets were acclimatized under greenhouse conditions to evaluate somaclonal variation.     

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.     

Cryopreservation of in vitro almond shoot tips by vitrification

Summary

Shoot tips of two almond scion cultivars, ‘Ne Plus Ultra’ and ‘Nonpareil 15-1’, and one almond/peach hybrid rootstock were successfully cryopreserved using a one-step vitrification technique. Three week old in vitro cultures were cold-hardened at 4°C on the multiplication medium (Murashige and Skoog for ‘Ne Plus Ultra’ and the hybrid rootstock; Almehdi and Parfitt for ‘Nonpareil 15-1’) for three weeks. Shoot tips, 2–2.5 mm long, were excised and precultured for 1 d at 4°C on the same basal medium, without plant growth regulators, supplemented with 0.7 M sucrose. After the preculture, the shoot tips were incubated in vitrification solution at 25°C for 45 min for the almond scion cultivars and 60 min for the hybrid rootstock, and then stored under liquid nitrogen (LN) for at least 3 d. After rapid thawing at 30°C, the shoot tips were washed with the appropriate liquid basal medium containing 1.0 M sucrose and then cultured on the same basal medium, solidified with agar, but excluding NH₄NO₃ or (NH₄)₂SO₄. Shoot regeneration was usually observed within 2–3 weeks. Survival after LN, recorded as the percentage of shoot tips that produced at least one new shoot four weeks after thawing, was 87.5, 60.0 and 72.5% for ‘Ne Plus Ultra’, ‘Nonpareil 15–1’ and the hybrid rootstock respectively. The one-step vitrification method is a promising simple technique for cryopreserving almond scion and rootstock shoot tips from in vitro cultures.     

地被月季‘Royal Bassino’高频再生体系的建立

 以地被月季‘RoyalBassino’为试材, 进行了组培快繁和外植体再生条件的研究。结果表明,以带有腋芽的茎段为外植体, MS中添加6-BA 1.5 mg/L和IBA 0.1 mg/L 为最适诱导培养基, 诱导出芽率100%; MS中添加6-BA 0.5 mg/L和IBA 0.05 mg/L可以获得最高增殖倍数(6.3) ; 而添加IBA 0.01 mg/L的1/2MS为最佳生根培养基, 生根率92%。试管苗经7 d驯化后, 移栽成活率在99%以上。分别以叶柄、叶盘和茎段为外植体进行再生培养, 以MS为基本培养基, 叶柄在添加噻苯隆(TDZ) 7μmol /L的诱导培养基中暗培养10 d后, 转接到添加6-BA 0.5 mg/L的再生培养基中光照培养约20 d, 可以获得57.1%的芽再生率。     

Investigations into inconsistent and low bud-grafting success in Acer platanoides ‘Crimson King’

Causes of variable and inconsistent bud-grafting success in chip-budded Norway maple (Acer platanoides) were investigated using the red-leaved scion ‘Crimson King’. Large differences in bud-take were associated with the year of budding, which over a 14 year period ranged from 22 to 94%. The ‘year effect’ could not be explained by weather, or by differences in budding success found between commercial sources of red-leaved scions. Neither was incompatibility between the ‘Crimson King’ scion and individual genotypes among the seedling rootstocks implicated, although there were reasons why at first sight incompatibility was thought to be present. The size of rootstock when planted was not correlated with subsequent budding success, although large differences in the quality of rootstock growth at budding were associated with large differences in bud-take, but these were confounded with year and with the fields used for the experiments. The ‘year effect’ was best explained by the field in which budding took place. One field gave consistently poor results, associated with a readily compacting soil, pointing to the strong possibility that bud-take is determined by conditions which affect shoot and/or root growth during the budding season or specifically while the union is forming in late summer. It was shown also that the presence of the scion bud on the chip-graft contributed to union formation, and that infection of the rootstock with the soil-borne verticillium wilt disease depressed bud-take, but this did not explain the ‘field effect’.     

Propagation in vitro of pear, Pyrus communis L., cultivars ‘William's Bon Chrétien’, ‘Packham's Triumph’ and ‘Beurré Bosc’

Shoot cultures were established from nodal explants of 3 pear cultivars by subculture on Murashige and Skoog medium containing mixtures of 3 cytokinins. Zeatin and 6·(γ,γ-dimethylallylamino)purine were both supplied at 10 μM and benzyladenine as follows: ‘William's Bon Chrétien’, 6 μM; ‘Packham's Triumph’, 8 μM; ‘Beurré Bosc’, 10 μM. Production of axillary shoots was greater by basal explants (4–6) comprising the remnants of the previous subculture than by apical explants (2–3) comprising the distal parts of extension shoots. Up to 80% of microcuttings of all cultivars formed roots in vitro with either γ-indolebutyric acid, 10 μM, or α-naphthaleneacetic acid, 10 μM, and approximately 50% of rooted microcuttings were successfully established as container-grown plants. Use of aseptic methods for producing own-rooted pears is discussed in relation to fruit growing and fruit improvement.     

Influence of FeEDDHA on in vitro rooting and acclimatisation of red raspberry (Rubus idaeus L.) in peat and vermiculite

Summary

We investigated the dependence of in vitro rooting and acclimatisation to greenhouse conditions on the source of iron used in the shoot multiplication and rooting media using five raspberry (Rubus idaeus L.) cultivars (‘Beskid’, ‘Canby’, ‘Malling Seedling’, ‘Norna’, and ‘Veten’). Ethylenediamine di-2-hydroxy-phenyl acetate ferric (FeEDDHA) in the rooting medium led to higher chlorophyll contents, earlier and more abundant rooting (8.7 vs. 5.3 roots per shoot), 30% higher fresh and dry weights, and thus higher quality microplants than ethylenediaminetetra-acetate ferric sodium (FeEDTA). Higher quality microshoots had a beneficial effect on acclimatisation (i.e., percentage survival and length of shoots) when the microplants were planted in a peat-based substrate; however, when planted in vermiculite, the initial differences disappeared during a 4 week-long growth period in a greenhouse.     

Root formation,bud growth and survival of ornamental shrubs propagated by cuttings on different planting dates

Summary

Semi-hardwood cuttings of Cornus alba ‘Sibirica’, Deutzia ‘Mont Rose’, Forsythia × intermedia ‘Lynwood’, Ligustrum vulgare ‘Liga’, Philadelphus × virginalis, Potentilla fruticosa ‘Goldfinger’, and Spiraea × vanhouttei were planted on seven dates from July to October in two years and at three locations to investigate the effect of planting date on root formation, axillary bud growth, and plant survival. Cuttings were planted directly in the field and covered with polyethylene. Generally, root formation, bud growth, and plant survival were similar both years and at the three locations. All species except Deutzia had relatively constant rooting percentages at planting dates until mid August. For all species rooting percentages declined from mid August to October. Except for Deutzia, plant survival the following spring was constant or decreasing with planting date. For all species except Potentilla axillary bud growth and survival of cuttings planted in late September or October tended to increase while rooting percentages continued to decrease. In all species there was a close relationship between axillary bud growth and survival. Results revealed that many roots per cutting accelerated axillary bud growth.     

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.     

Root induction in three species of bamboo with different rooting abilities

Rooting of in vitro axillary shoots from adult field culms of Bambusa atra, Dendrocalamus giganteus and D. hookeri and of juvenile seedling shoots of D. giganteus were investigated. B. atra rooted spontaneously without exogenous auxin during axillary shoot proliferation, while both Dendrocalamus species rooted only on transfer to rooting media with indole-3-butyric acid (IBA). D. giganteus required coumarin, an auxin protector, in addition to IBA. Rooting in adult shoots of D. giganteus was lower (45.6%) than that in the juvenile shoots (96.7%) but adult shoots of D. hookeri rooted well (88.9%). A pretreatment with thidiazuron (TDZ: N-phenyl-N-[(1,2,3-thidiazol-5-yl)urea]) induced development of axillary buds and subsequently 95% rooting in adult D. giganteus shoots but had no significant effect on rooting in juvenile shoots of D. giganteus or D. hookeri. Pretreatment with tri iodobenzoic acid (TIBA) arrested shoot and bud development leading to very low or no rooting in all three species.     

High-frequency somatic embryo production from unfertilized ovules of grapes

Somatic embryos were produced in large numbers from cultured, unfertilized ovules of Vitis vinifera, cultivars ‘Cabernet Sauvignon’ and ‘Grenache’, and of the hybrid grape ‘Gloryvine’ (V. vinifera × V. rupestris). Callus of nucellar origin was produced by culturing the ovule explants initially in Nitsch medium containing 5 μM 2,4-dichlorophenoxy acetic acid (2,4-D) or 5 μM β-naphthoxyacetic acid (NOA) plus 1 μM benzyladenine (BA), and then in a medium supplemented with NOA (10 μM) plus BA (1 μM). Embryos were produced when callus was transferred to basal medium containing no auxin or cytokinin. Secondary and tertiary embryos were formed by budding of primary somatic embryos. Prolonged culture of ovular callus in NOA (10 μM) plus BA (1 μM) gave rise to cell aggregates which developed into free-floating roots when transferred to basal medium.     

Regeneration of plants from Fraxinus pennsylvanica hypocotyls and cotyledons

An adventitious shoot regeneration and rooting protocol was developed for green ash (Fraxinus pennsylvanica) seedling explants. The best regeneration medium for freshly isolated hypocotyls and cotyledons was Murashige and Skoog (MS) supplemented with 13.3 μM 6-benzylaminopurine (BA) plus 4.5 μM thidiazuron (TDZ), and 22.2 μM BA plus 4.5 μM TDZ, respectively. Seventy-six percent of hypocotyl segments and 24% of cotyledon segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 2.7 ± 0.5 and 2.3 ± 1.3, respectively. The effect of in vitro-germinated seedling age on adventitious shoot regeneration from hypocotyl and cotyledon explants was also studied. Results showed that hypocotyl and cotyledon explants from freshly isolated embryos exhibited a higher organogenesis potential than 4–15-day-old explants. Adventitious shoots from hypocotyls and cotyledons were established as proliferating shoot cultures following transfer to MS basal medium with Gamborg B5 vitamins supplemented with 10 μM BA plus 10 μM TDZ. A high rooting percentage (73–90%) was achieved when in vitro shoots were rooted on woody plant medium (WPM) containing 4.9 μM indole-3-butyric acid (IBA) and IAA (0, 2.9, 5.7, or 8.6 μM) with a combination of 10-day dark culture period followed by a 16-h photoperiod. The highest rooting (90%) of adventitious shoots or the number of roots per shoot (3.0 ± 1.0) was obtained on WPM with 4.9 μM IBA plus 5.7 μM IAA. Rooted plants were successfully acclimatized to the greenhouse and 100% survived after overwintering in cold storage. This regeneration system using hypocotyls and cotyledons provides a foundation for Agrobacterium-mediated genetic transformation of F. pennsylvanica for resistance to the emerald ash borer.     

Propagating palms in vitro with special emphasis on the date palm (Phoenix dactylifera L.)

Tissue-culture methods are described for the vegetative propagation of several palm species either through shoot tip culture or plantlet differentiation via embryogenic callus. The influence of explant size, medium composition and physical environment required for the establishment of palm shoot tips in vitro was determined. Date palm (Phoenix dactylifera L.) seedling shoot tips of various sizes were cultured in either liquid or agar modified Murashige and Skoog (MS) medium containing 0.0–1.0 mg 1^?1 α-naphthaleneacetic acid (NAA) and 0.0–15.0 mg 1^?1 benzyladenine or N⁶-(Δ²-isopentenyl) adenine (2iP) in order to enhance shoot growth and induce axillary budding. Satisfactory date palm shoot tip growth and proliferation was obtained from explants that were 3 mm in length, consisting of the apical meristem region and 2–5 adjacent leaf primordia. Optimum shoot tip development and axillary budding was obtained by initially establishing explants on an agar medium for 2 weeks, then transferring to a liquid medium. Shoot tips from several palm species were cultured on MS media containing 100 mg 1^?1 2,4-dichlorophenoxyacetic acid (2,4-D), 3 mg 1^?1 2iP and 3 g 1^?1 activated charcoal, or on MS medium containing 1 mg 1^?1 NAA and charcoal, to determine their morphogenetic responses in vitro. Shoot tips of Metroxylon sp., Phoenix canariensis Hort. ex. Chabaud., P. dactylifera ‘Khalasa’, ‘Thoory’ and ‘Zahidi’, and P. roebelenii O'Brien planted on medium with 2,4-D and 2iP initiated callus, asexual embryos and free-living plantlets after 4–8 months in culture. Shoot tips from Erythea edulis S. Wats., P. canariensis, P. dactylifera ‘Khalasa’, Thoory' and ‘Zahidi’, Washingtonia filifera Wendl. and W. robusta Wendl. cultured on medium containing NAA developed into plantlets with well-developed leaves and adventitious roots within 2–6 months from the time of planting. In some cases, cultured date palm shoot tips gave rise to axillary buds.     

In vitro propagation of ‘Arka Ravi’ chrysanthemum on growth regulator-free medium harbouring endophytic bacteria

Summary

To develop a micropropagation protocol for the elite chrysanthemum cultivar, ‘Arka Ravi’, in vitro cultures were established using surface-sterilised nodal microcuttings (1.0 – 1.2 cm) on semi-solid MS medium. Microbial contamination was observed in 22% of cultures during the initiation phase. Cultures that were devoid of obvious contamination were transferred to culture bottles containing MS medium supplemented with 30 g l^–1 sucrose, 2.5 g l^–1 Phytagel^® and either benzyl adenine (BA) or kinetin (KIN) supplied at 0, 1, 5, 10, or 20 µM, and were monitored, over eight in vitro passages, for their growth and microbial association. Shoot-tip and nodal microcuttings yielded a single shoot, coupled with rooting, in medium devoid of BA or KIN which was the best medium for continuous micropropagation. Rooting was inhibited with increasing concentrations of BA or KIN, and one or more shorter shoots with condensed internodes were induced, resulting in low rates of propagation. Culture indexing (i.e., testing the medium and tissue from visibly clean cultures using enriched bacteriological media) revealed quiescent endophytic bacteria associated with 80 – 100% of such cultures. Three distinct colony morphotypes were isolated and were identified as Ralstonia, Enterobacter, and Methylobacterium spp., based on their 16S rRNA gene sequences. These endophytes did not interfere with normal micropropagation, but tended to grow actively and outgrow older cultures, especially at higher cytokinin concentrations. Stable micropropagation of ‘Arka Ravi’ chrysanthemum for ≥ 2 years, with their resident endophytic bacteria in a covert form, was achieved on basal MS medium with > 90% shoot growth and rooting, a four-to-five-fold propagation rate at each 2 – 3 week sub-culture cycle, and with > 90% establishment of rooted plantlets ex vitro. These results suggest that in vitro cultures of chrysanthemum often harbour endophytes with no obvious indications of their presence or with possible hidden effects during micropropagation.     

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

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

Evaluation,tissue culture propagation,and dissemination of ‘Saba’ and ‘Pelipita’ plantains in Costa Rica

Fruit characteristics of the disease-resistant bluggoe-type (ABB) cooking bananas (Musa × paradisiaca L.) ‘Saba’ and ‘Pelipita’ were compared with those of the susceptible ‘Currare’ (‘Horn’). Yields of ‘Saba’ and ‘Pelipita’ were similar to those of ‘Currare’; however, ‘Saba’ and ‘Pelipita’ yielded fewer hands with a greater number of small fruit when compared with ‘Currare’. Shoot-tip cultures of both clones were readily initiated on a modified Murashige and Skoog (MS) basal medium supplemented with N⁶-benzyladenine (BA) in combination with indole-3-acetic acid (IAA). Propagation cultures were initiated by splitting shoot tips along their longitudinal axis and re-culturing the individual pieces to basal medium supplemented with 5 mg l^?1 BA. Transfer of axillary shoots to hormone-free medium resulted in rapid and extensive root formation. Plantlet survival after transfer to methyl bromide-treated soil exceeded 90%. Establishment in the field was achieved following procedures normally used for vegetative propagation of this crop. Trial plantings of in vitro propagated ‘Saba’ and ‘Pelipita’ were established in various provinces of Costa Rica for grower evaluation and for future comparison of growth and reproductive development with plants of these and other cultivars propagated from corms.     

Direct and callus-mediated regeneration of Curcuma soloensis Valeton (Zingiberaceae) and ex vitro performance of regenerated plants

Protocols are outlined for the regeneration of Curcuma soloensis, an attractive tropical ornamental plant, from young vegetative bud explants. We used both direct and callus-mediated regeneration techniques to produce material suitable for mass propagation and the development of transgenic plants. During direct plantlet propagation, the presence of thidiazuron (TDZ) in the growing medium induced more than three times as many shoots as 6-benzylaminopurine (BA), with a mean of 18.7 shoots per explant on MS medium containing 2.5 μM TDZ compared to 5.0 shoots with 40 μM BA. Subsequently, the shoots rooted readily on MS basal medium that was free of plant growth regulators. During indirect plantlet regeneration, TDZ combined with BA and 2,4-dichlorophenoxyacetic acid (2,4-D) had significant effects on embryogenic callus induction and multiplication. The frequency of callus formation was 91.1% for explants cultured on MS basal medium supplemented with 2.5 μM TDZ, 2.0 μM BA and 1.2 μM 2,4-D. On average 7.1 shoots were produced per callus mass cultured on MS medium supplemented with 2.5 μM TDZ, 9.0 μM BA and 1.2 μM naphthaleneacetic acid (NAA). Regenerated shoots were transferred to MS medium supplemented with 2.5 μM TDZ, to produce multiple shoots. In vitro cultured plantlets readily acclimatized to greenhouse conditions, showing 100% survival rates in a sphagnum, perlite and sand (1:1:1) medium. These plants were transplanted into pots or planted in the field. The ex vitro acclimated plants grew vigorously and produced showy inflorescences 5–6 months after planting. The high-frequency of shoot multiplication and rapid flowering of tissue-cultured plants indicate that C. soloensis has great potential in the floricultural market.     

Improvement of in vitro micropropagation of Lilium oriental hybrid ‘Casablanca’ by the formation of shoots with abnormally swollen basal plates

Bulb scales of Lilium oriental hybrid ‘Casablanca’ were cultured on MS medium supplemented with cytokinins (kinetin, BA (benzyladenine), TDZ (thidiazuron)). The basal part of bulb scales swelled and formed adventitious shoots with foliage small, leafy bulb scales and abnormally swollen basal plates on the media with cytokinins. Shoots were formed rapidly from the basal parts of bulb scales and became shoot clusters. The medium containing 2.2 μM BA was most favorable in the shoot formation from bulb scales. Cutting shoots into small segments (7–8 mm wide × 12–15 mm length) were cultured on media containing BA and IAA (indole acetic acid) and the segments regenerated many new shoots and formed shoot clusters. The shoot section to shoot cluster cycle method improved adventitious shoot proliferation. The media supplemented with 4.4 μM BA and 5.7 μM IAA. or 8.9 μM BA and 0.6–2.9 μM IAA were effective in allowing the proliferation of shoots from shoot segments under light condition. Sucrose and activated charcoal (AC) improved bulblet growth. Bulblet growth was effectively performed on MS medium containing 60 g/L sucrose. Bulblet growth was also improved by the supplement of AC. The medium with 2.0 g/L AC was most effective for bulblet growth. Normal bulblet growth was stimulated more by the culture of shoots than that of bulb scales. Bulblet weight from shoots reached to an average of over 1100 mg of a bulblet after 3 months in culture on MS medium containing 60 g/L sucrose and 2 g/L AC. Most of the bulblets produced by this method generated stems with several leaves in the green house, after cold treatment at 5 °C for 3 months.     

Nutritional and cultural factors affecting rooting of papaya (Carica papaya L.) in vitro

Summary

Experiments were conducted to optimize nutritional and cultural requirements for initiation and growth of roots on papaya in vitro. Axillary shoots were obtained from plants which had been sub-cultured monthly for two years. Root initiation was enhanced when 1 to 2 mm of stem base was removed and shoots were growing actively before transfer to the rooting medium. Decreasing daylength during incubation from 24 h to 12 h promoted root initiation. Within the day temperature range of 22 to 29°C, optimum rooting occurred at 27°C and higher temperatures produced higher mean root weights per shoot. High concentrations of growth factors and the absence of sucrose in the medium both reduced root initiation, however, varying the concentration of sucrose and removing growth factors affected mean root weight per shoot. All media contained a modified de Fossard et al. (1974) basal medium plus 10 μM IBA.