Standardization of Rootstocks of Mango (Mangifera Indica L.)

Mango cuttings could be induced to form roots provided IBA at 5,000 p.p.m. in lanolin was applied at the base of shoots after removing a ring of bark while they were still attached to the mother tree.

Cuttings taken from one-month-old seedlings gave the highest percentage of rooting and establishment, but with increasing age of the mother plants the rooting was considerably reduced. The rooting of cuttings of different age groups could be improved by using shoots forced into vigorous growth by hard pruning. Younger plants responded better to such treatment than older ones.

Cuttings from etiolated shoots rooted better than those from non-etiolated, but etiolation was less effective in promoting rooting than invigoration by hard pruning. Etiolation and invigoration combined gave the highest percentage of rooting and establishment in all age groups.     

Effect of gibberellic acid (GA3) on elongation and rooting of ‘St. Julien A’ rootstock in vitro

‘St. Julien A’ (Prunus instititia L.) rootstock was induced to proliferate shoots on a modified half-strength Murashige and Skoog (MS) medium. Cultures treated with 12.5 mg l^?1 gibberellic acid (GA₃) produced elongated shoots suitable for rooting. Elongated shoots were placed in media with indolebutyric acid (IBA) or indole-3-acetic acid (IAA) with or without a 16-day dark incubation. Light (16-h photoperiod) inhibited rooting. IAA (4 mg l^?1) was ineffective in promoting rooting. Rooting was best when shoots were incubated in the dark with IBA (4 mg l^?1). GA₃ was deleterious to shoots, causing chlorosis and apical die-back. Light regime interacted with auxin treatments in affecting shoot condition. Shoot condition was better on shoots treated with IBA and dark-incubated; while those treated with IAA were better when light-incubated.     

Micropropagation of Prunus scoparia,a Suitable Rootstock for Almond under Drought Conditions

Prunus scoparia is a wild deciduous shrub, usually living on dry calcareous soils of the rocky mountains and has been used as a grafting rootstock for domesticated almonds to provide drought resistance. In the current study, micropropagation ability of P. scoparia was investigated using cytokinin and auxin. Uniform nodal shoot pieces (3–5 cm in length) of seedlings were used as explants. The explants were disinfected with 10% sodium hypochlorite solution. For adventitious shoot induction and proliferation, Murashige and Skoog (MS) media containing 7.00 g/l agar and 30.00 g/l sucrose containing five concentrations of benzyl adenine (BA) (0.00, 0.50, 1.00, 2.00, and 4.00 mg/1) and also containing six concentrations of Thidiazuron (TDZ) (0.00, 0.50, 1.00, 2.00, 5.00, and 7.00 mg/1) were compared. For rooting, in vitro shoots (2–3 cm) were transferred into ½ MS medium supplemented with 30 g/l sucrose, 7.50 g/l agar, and different concentrations of IBA (0.00, 0.25, 0.50, and 1.00 mg/l) and NAA (0.00, 0.25, 0.50, and 1.00 mg/l). Based on the results obtained for shoot proliferation, only 2.00 and 4.00 mg/l BA and 2.00 mg/l TDZ concentrations generated shoots, while other treatments did not show shoot proliferation. Among the three treatments that generated shoots, the best results for shoot number, leaf number, and leaf color quality were observed in media containing 2.00 mg/l TDZ. Based on the results obtained for rooting, the effect of IBA concentrations on the rooting percentage, root number, and root length was significant. Among IBA concentrations, only 0.50 mg/l IBA induced rooting, while there was no rooting in the media containing other IBA concentrations. None of the NAA concentrations showed rooting. In conclusion, MS culture medium supplemented with 2.00 mg/l TDZ and ½ MS culture medium supplemented with 0.50 mg/l IBA are suggested for in vitro shoot proliferation and rooting of P. scoparia, respectively. The results presented herein could be used for in vitro selection and micropropagation of P. scoparia.     

Production of high quality Ardisia plants by stem tip cuttings

To produce commercially acceptable Ardisia plants, stem tip cuttings from mature plants were rooted and forced in greenhouses. Ten centimeter long cuttings were either treated with 200 ppm 1-naphthalene acetic acid (NAA) for 2 h, 2000 ppm indole-3-yl-butyric acid (IBA) for 10 s, or 0.5 and 1.0% IBA powder prior to sticking them in the rooting medium. Rooting percentage at 45 days exceeded 76% with 2000 ppm IBA treatment which was a significant increase over non-treated control. Rooted cuttings developed into three types of plants: those forming only vegetative shoots without flowers, those forming reproductive shoots with flowers, and those forming both vegetative and reproductive shoots. The ideal plant produced only vegetative shoots when rooted cuttings were transplanted into pots. About 50% rooted cuttings were forced to finish, producing 31 or 40% of high quality plants when rooted cuttings with vegetative shoots were grown in a greenhouse (GH) at temperatures higher than 21/19 °C (day/night) in 1995 or 21/18 °C GH in 1997, respectively. This method shortened the total production time to less than 2 years as compared to 4 years when starting from seeds.     

Factors affecting tissue culture of Damask rose (Rosa damascena Mill.)

The present study was conducted to evaluate the regeneration ability of Damask rose. Single-node explants were surface sterilised with 10% chlorox for 15 min and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N⁶-benzyladenine (BA) or kinetin (Kin) separately or in combination with different concentrations of indole-3-butyric acid (IBA). Combination of 2.5–3 mg/l BA and 0.1 mg/l IBA was the most suitable treatment for proliferation. In vitro derived shoots were subcultured four times on the fresh medium within a 4-week period. Other treatments such as explant orientation (horizontal, vertical and oblique) and explant wounding were also examined but did not affect shoot multiplication rate significantly. Several experiments were carried out to stimulate in vitro rooting of Damask rose. Application of different media such as MS, 1/2 MS, 1/3 MS and 1/4 MS with different concentrations of indole-3-acetic acid (IAA), IBA and naphthaleneacetic acid (NAA) did not produce satisfactory results. Quick-dip method using sterilised 0–2000 mg/l IAA, IBA and NAA solutions was also studied. Other treatments such as using various concentrations of abscisic acid (ABA) in combination with various concentrations of IAA, IBA and NAA, and using various concentrations of sucrose and agar did not produce any roots. The best treatment for rooting of shoots was 2.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for 2 weeks in MS medium and then transferring the explants to MS medium without any growth regulator. Plantlets were acclimatised in a soil mixture consisting of peat moss and sand 1:1 (v/v) and successfully transferred to the greenhouse after 3 weeks.     

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.     

De novo differentiation of shoot buds from leaf-callus of Dianthus caryophyllus L. and control of hyperhydricity

A method is described for producing de novo shoots from leaf derived callus of carnation (Dianthus caryophyllus L.). Plants were regenerated in four steps, viz. callus induction, shoot regeneration, removal of hyperhydricity from regenerated shoots and root development. Callus induction medium contained 2,4-D and BAP. Shoot buds were formed when the callus was further subcultured on 2,4-D- and BAP-containing medium, or MS medium without any growth regulators. The shoots so formed were hyperhydric, bushy in appearance with reduced stem length and watery leaves. The normal conformation of shoots was restored by culturing the hyperhydric shoots onto medium supplemented with GA₃ and bactopeptone. The recovered shoots were rooted on MS medium added with NAA (1 mg/l) or IBA (2 mg/l). Regenerated plants with well-developed root and shoot systems were successfully transferred to field conditions after initial acclimation.     

In vitro micropropagation from plumular apices of Turkish cowpea (Vigna unguiculata L.) cultivar Akkiz

Multiple shoot formation from the plumular apices excised from mature embryos of cowpea cv. Akkiz was obtained after pulse treatment with 10 mg/l BAP for 5 days followed by culture on MS medium containing 0.25, 0.50, 0.75 and 1.00, 1.25 mg/l BAP – with or without 0.10 mg/l NAA. Callus induction and shoot regeneration was recorded on all cultures containing BAP with or without NAA. However, inclusion of 0.1 mg/l NAA had positive effect on callus diameter and shoot length. Maximum mean number of 7.11 shoots per explant were obtained on MS medium containing 1.00 mg/l BAP. Longer shoots were recorded on MS medium containing various concentration of BAP+ 0.1 mg/l NAA compared to those containing various concentrations of BAP singly. All shoots cultured on MS medium containing 1 mg/l BAP were rooted on MS medium containing 0.50 mg/l IBA. Rooted plants were acclimatized at room temprature in soil contained in pots. All plants flowered and set seeds in the greenhouse after 3 months.     

In vitro vegetative multiplication of Fuchsia hybrida

Rapid development of axillary buds from shoot-tips and nodes of 18 cultivars of Fuchsia hybrida was obtained on solid Murashige and Skoog medium with BAP (1 mg l^?1 and an auxin (0.1 mg l^?1). NAA as the auxin appeared to be more active than IAA or IBA. Vegetative shoots were subsequently isolated and developed up to 15 supplementary axillary shoots on the same solid medium. Agitated and non-agitated liquid media of the same composition were less effective. One-cm long shoots could be rooted in 20 days in the presence of IBA before being transferred to soil.     

Hormone effects on in vitro proliferation and rooting of Grevillea explants

Explants of Grevillea rosmarinifolia A. Cunn. and Grevillea×semperflorens F.E. Brigs ex Mullig., obtained from vegetative stem segments, were grown on a modified MS medium, which was supplemented with different contents of cytokinins (BA or K) to evaluate optimal conditions for bud proliferation. Auxins (NAA or IBA) were tested to evaluate rooting.

The multiplication rate was often higher in G. rosmarinifolia than in G.×semperflorens. Explant activity varied significantly depending on the cytokinin type and its concentration. The addition of cytokinins to the culture medium often promoted an increase in the number of shoots produced by each propagule, but at the highest BA concentration, shoot differentiation was inhibited in Grevillea×semperflorens. Only the higher concentrations studied (5–10 mg/l) produced better results with K, whereas BA was more effective at 1–2 mg/l. Rooting efficiency of the young shoots was lower in G. rosmarinifolia than in G.×semperflorens. IBA promoted root development in both species and was more effective at the highest concentration (1 mg/l). NAA gave better results at low concentrations (0.2 mg/l) in G. rosmarinifolia, while it had slight or negative effect on rooting of G.×semperflorens.     

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。     

Globe artichoke plants obtained from shoot apices through rapid in vitro micropropagation

A rapid method of in vitro propagation for globe artichoke (Cynara scolimus L.) is described. Shoot apices and subterminal stem segments were cultured on modified Murashige and Skoog (1962) medium (MS) with NaH₂PO₄ (50 mg/l), m-inositol (100 mg/l), L-tyrosine (100 mg/l), adenine (40 mg/l), indoleacetic acid (IAA, 0.5 mg/l), kinetin (K, 10 mg/l). sucrose (4%) and agar (0.7%) at pH 5.5. On this medium, a high number of proliferating shoots was obtained. The number of these shoots was increased and their overall development improved by sub-culturing on a MS medium with a reduced concentration of K (5 mg/l) and without the additional amount of sodium dihydrogen phosphate. In these conditions, a 4.5 rate of shoot proliferation was reached after 3 weeks.To induce rooting, the proliferated shoots were transferred to a medium containing half MS, thiamine-HCl (1 mg/l), m-inositol (100 mg/l), ascorbic acid (10 mg/l), naphthaleneacetic acid (NAA, 2 mg/l), sucrose (2%), agar (0.7%) at pH 5.5. In these culture conditions about 84% of plantlets rooted.Cytological analyses performed on root tips of 20 randomly chosen plantlets showed that all the analysed plants contained the diploid number of chromosomes. The plantlets were successfully transferred to soil and the method described seems to be suitable for rapid propagation of globe artichoke.     

促进马扎德甜樱桃砧木组培苗生根的研究

以甜樱桃砧木马扎德(M azzard)的组培苗为试材,研究促进其生根的方法。结果表明,最佳生根培养基为1/2 MS 0.3mg/L IBA。将分化组培苗转入该培养基,移至温室中黑暗处理10天后,在1500~4000 lx自然光下培养,生根率可达93.5%。对瓶内未生根的幼苗,用1000mg/L IBA浸泡60秒后扦插到温室育苗盘中,1个月后生根率可达80%以上。     

In vitro culture of cotyledon tissue of Castanea sativa Mill

Establishment of callus tissue culture from cotyledon fragments of Castanea sativa Mill. is described. The explants were grown on various combinations of auxins (IAA, IBA, NAA, 2,4-D) and cytokinins (K, BA) to induce callus and regeneration of organs. The best growth occurred on basal medium with 2,4-D (1 and 10 mg/l) plus both K or BA (0.5 mg/l). Root regeneration was achieved with both IBA and NAA (10 mg/l) supplemented with K or BA.     

黑莓试管苗叶片植株的再生

以黑莓带芽茎段试管苗叶片为外植体,诱导形成不定芽并进一步形成再生植株。在MS+TDZ2.0mg/L的培养基中叶片不定芽最高分化频率达93.7%(6.52不定芽/外植体),在不定芽伸长培养基BA0.5mg/L上,分化完全的不定芽能够长大伸长,当其高度达到4cm时,切下转接于IBA为0.3mg/L培养基上诱导生根,生根率100%。     

The use of CPPU for efficient propagation of pineapple

The use of forchlorfenuron (N-(2-chloro-4-pyridyl)-N′-phenylurea) (CPPU) for efficient propagation of pineapples was investigated. About 85% of axillary buds can be forced to sprout by soaking defoliated stem pieces (12 cm in length) in a 2.5 or 5.0 mg l⁻¹ CPPU solution for more than 3 h. The use of old stems taken from the third or fourth ratoon plants had the advantage of less liability to fungal decay, as compared to young stems from the first crop plants. The CPPU treatment combined with the removal of shoots from stems at monthly intervals significantly increased the number of shoots per stem piece (about 15 shoots per piece at 5.0 mg l⁻¹ CPPU), and resulted in a more uniform shoot size (the percentage of shoots within a range of 5–15 cm in length was about 90% at 5.0 mg l⁻¹ CPPU). The rooting of shoots was easily promoted within 1 month by treating the basal portion of shoots with 20 mg l⁻¹ indole-3-butyric acid (IBA) for 15 min. The CPPU method required about 5 months for plantlet propagation. From these results, we found that pineapple plantlets could be efficiently propagated by the following method: (1) soaking defoliated stems in a 2.5–5.0 mg l⁻¹ CPPU solution for more than 3 h; (2) harvest of developed shoots from the stems at regular intervals; and (3) promotion of rooting on the shoots at 20 mg l⁻¹ IBA for 15 min.     

Adventitious shoot regeneration from in vitro leaves of adult Camellia reticulata

Experiments were conducted with Camellia reticulata cv. Captain Rawes to develop an adventitious regeneration system. Leaves were taken from axillary shoot proliferation cultures in WPM medium that had been established from mature trees. They were sectioned, and then plated in Petri dishes on media containing various combinations of cyto- kinins and auxins; the best response was induced by 2 mg I“1 BA + 1 mg I-1 IBA. The shoots obtained were multiplied by axillary branching on WPM + 2 mg I-1 BA + 2 mg I-1 Z + 2 mg I-12iP + 0.01 mg I-1 IBA. There was no significant difference in multiplication coefficient (the product of the proportion of explants forming axillary shoots and the mean number of new segments per explant) between shoots of adventitious and axillary origin, but there was between the various types of explant used in the multiplication stage: shoot tips (ST1) and nodal segments (ns) of harvested shoots longer than 14- 15 mm, and whole harvested shoots 7-10 mm long (ST2). The best results were achieved with ns and ST2 explants. Shoots of adventitious origin rooted very poorly in comparison with those of axillary origin under the same culture conditions.     

Rooting stem cuttings of Chinese chestnut

Rooting of Chinese chestnut (Castanea mollissima Blume.) stem cuttings following treatment with indole-3-butyric acid (IBA) was investigated. Tip and basal cutting from vigorous epicormic shoots and terminal shoots with 2–3 cm of the previous year's wood were taken from mature regions of trees approximately 40 years of age. Cuttings were dipped for 5 s in an aqueous solution of either 3 or 6 g l^?1 IBA. Rooting occurred only in the basal softwood cuttings. Average rooting of 33.5, 5.0 and 1.6% for ‘AU-Leader’, ‘AU-Homestead’ and ‘AU-Cropper’, respectively, was obtained using the 3 g l^?1 IBA solution, and 35.0, 6.7 and 3.3%, respectively, using the 6 g l^?1 IBA solution.     

桃幼胚下胚轴高频植株离体再生

以桃品种京艳和晚蜜不同发育阶段合子胚下胚轴为外植体进行离体培养,用6-BA、CPPU和TDZ等细胞分裂素诱导下胚轴直接再生不定芽试验。结果表明,花后70d下胚轴为诱导的最佳材料,供试细胞分裂素中,诱导活性为TDZ>CPPU>6-BA,苯脲类细胞分裂素TDZ5mg/L与NAA0.01mg/L配合对下胚轴再生具有极强的诱导作用,京艳诱导再生率高达95.2%,平均不定芽数达每外植体25.3个。京艳的诱导率显著高于晚蜜。IBA 0.5mg/L适于诱导再生幼茎的生根,生根率和移栽成活率分别达到92.6%和65.5%。组织学观察表明不定芽起源于皮层薄壁细胞,属直接再生方式。     

Micropropagation of gooseberry, Ribes grossularia

A method for rapid micropropagation of gooseberry (Ribes grossularia cultivar ‘Hinnonmäki Yellow’) was established. The optimum concentration for shoot production was obtained at 2.2 μM BAP and 0.05 μM IBA with MS nutrient medium. The Lepoivre salt medium with no hormones was used for elongation of shoot clusters, and in this medium about 50% of the shoots formed roots. For both rooted and unrooted shoots, 100% survival was obtained after transfer to soil. It was possible to store shoots in vitro for 130 days without any sub-culture and with 100% survival if the temperature was increased successively after cold storage.