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.     

Propagation characteristics of Eucalyptus globulus Labill. ssp. globulus stem cuttings in relation to their original position in the parent shoot

The rooting ability of one-node leafy cuttings of Eucalyptus globulus decreased rapidly with distance from the (decapitated) apical node. The young and immature leaves close to the shoot apex usually promoted rooting whereas the apex itself had little effect or was detrimental. In apical cuttings with an equivalent complement of leaves, increasing stem volume (and woodiness at the base of the cutting) slightly reduced rooting. The swelling of the stem at the base of shoots from coppice or potted plants also reduced rooting when present at the base of the cutting, whereas the inclusion of the root/shoot junction of seedlings promoted rooting. Cuttings from small seedlings rooted well and the roots sometimes emerged directly from the stem, whereas in non-seedling cuttings, roots nearly always arose from callus. The survival of cuttings in the propagation environment varied between trials. In apical cuttings with the same leaf complement, survival tended to increase with woodiness at the base of the cutting, but in contiguous one-node cuttings this tendency was overcome by the leaves, the soft apical nodes surviving better than the more proximal woodier nodes.     

Micropropagation of the strawberry tree,Arbutus unedo L.

Actively growing shoots of potted greenhouse-grown strawberry tree (Arbutus unedo L.) were initially sterilised and established in basal woody plant medium containing 11.1 μM BA. Optimum shoot proliferation was achieved on a basal WPM containing MS vitamins, sucrose, agar and 22.2 μM BA. Microshoots rooted successfully in basal in vitro medium containing 10 μM IBA or IAA, but their survival rate during acclimatisation was low. Addition of a mixture 1 part peat:4 parts perlite in the basal in vitro rooting medium (1:1 v/v) containing 10 μM IAA resulted in high rooting percentage and plantlets with branched roots. These plantlets were successfully acclimatised. This novel rooting medium can be exploited further due to its potential in commercial applications.     

Depressing Effects of Virus Infection on Adventitious Root Production in Apple Hardwood Cuttings

Adventitious rooting in dormant hardwood cuttings of MM. 106 apple rootstock was depressed by latent infection with five viruses, viz. rubbery wood, stem pitting, epinasty and decline, chlorotic leaf spot and platycarpa scaly bark viruses.

Both the number of rooted cuttings and the number of roots per rooted cutting were reduced, and fewer unrooted cuttings remained alive, thus representing a further potential loss.

The production of shoots suitable for use as cuttings was decreased from infected hedges, as was the ability of cuttings subsequently to become established and grow in the nursery.     

Effects of time of collection,growing-conditions of mother plants and growth regulators on rooting of cuttings of Telopea speciosissima (Proteaceae)

Terminal stem cuttings of Telopea speciosissima were collected from two locations at monthly intervals and treated with indolebutyric acid (IBA) in combination with benomyl. Cuttings taken in the first half of spring, at the beginning of vegetative growth, gave the highest percentage of rooted cuttings and the greatest length of adventitious roots per rooted cutting. Cuttings taken from the environment most suited for the growth of the mother plants also produced the highest percentage of rooted cuttings, but their root length did not differ significantly between the two locations. Pre-treatment of cuttings with 4000 p.p.m. IBA (concentrated solution dip method) initially gave the highest percentage of rooted cuttings and longest root length, but there was evidence for a delayed toxicity to IBA applied at this level. Pre-treatment with 2000 p.p.m. IBA is therefore recommended.Benomyl increased the percentage of rooted cuttings, but did not affect root length, possibly due to its fungicidal rather than cytokinin activity.     

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.     

Somatic embryogenesis and plant recovery from callus of ‘Nabali’ Olive (Olea europea L.)

Regeneration via somatic embryogenesis from callus was studied in ‘Nabali’ olive (Olea europea L.). Among different explant sources (leaf blades, leaf petioles, hypocotyls of germinated seeds and roots of germinated seeds), roots gave the highest (46%) callus induction. Somatic embryogenesis was induced from root callus on embryogenesis induction medium (EIM) containing 5.0 μM 2,4-D, 0.5 μM kinetin and 5.0 μM NAA in darkness. Embryo regeneration was studied by transferring the callus from EIM to embryogenesis expression medium (EEM) containing different concentrations (0.0, 5.0, 10.0 and 15.0 μM) of 2-isopentenyladenine (2iP), BA, thiadiazuron (TDZ), zeatin or kinetin. Among the tested concentrations, 2iP at 10.0 μM outperformed the other growth regulators. 2,4-D at 5.0 μM in the EIM was satisfactory for embryogenesis induction. Sucrose at 0.2 M evoked higher embryogenesis than any other concentration of fructose and glucose in EIM, while sorbitol and mannitol at 0.1, 0.2 or 0.3 M reduced embryogenesis significantly and inhibited it totally at 0.4 M. Somatic embryos were rooted by transferring them to hormone-free medium (HFM). About 85% of embryos converted to rooted plantlets, 5% showed secondary embryogenesis and 10% were not developed and died. Rooted plantlets gave 95% survival when acclimatized ex vitro. Acclimatized plantlets developed into whole plants in the greenhouse and they were phenotypically similar.     

Field performance of highbush blueberries (Vaccinium × corymbosum L.) cv. ‘Herbert’ propagated by cuttings and tissue culture

‘Herbert’ highbush blueberry (Vaccinium × corymbosum L.) plants propagated by softwood cuttings (HT) and obtained by micropropagation (TC) of axillary (AX) and adventitious (AD) shoots of 1-year-old in vitro cultures or 11-year-old cultures (SH) were compared. Propagation methods exerted significant influence on nursery and field performance of blueberries. Cutting-derived HT plants grew more slowly, produced significantly less and shorter shoots and were more variable than micropropagated plants. However, the majority of HT plants developed flowers 1 year earlier, flowered more abundantly, bore significantly larger berries than TC plants. There was no clear difference between AX and AD plants. SH-derived plants had smaller berries with the fewest seeds compared to AX and AD-obtained plants. This reveals that culture age had more significant influence than shoot source for the variation observed among micropropagation systems. The present study underlines the necessity of frequent establishment of in vitro cultures of highbush blueberry and carry them out by limited number of passages.     

GL生根剂对扶桑插条生根及碳水化合物分配的影响

用ＧＬ生根剂（ＩＢＡ１０００ｍｇ／Ｌ＋粉锈宁１５０ｍｇ／Ｌ＋ＰＰ３３３２ｍｇ／Ｌ）５０ｍｇ／Ｌ浸泡扶桑插条基部２４小时，提高了插条的生根率、生根数、根干重，并扩大了生根范围。插条叶光合速率高于对照８２．８％，分配到叶的１４Ｃ－光合产物比对照少，而分配到茎和不定根的光合产物高于对照。插条叶和不定根中的可溶性糖、淀粉和纤维素含量低于对照，并随处理后天数的延长逐渐减少，木质素含量增加；茎和插条基部的可溶性糖含量高于对照，淀粉和木质素含量低于对照     

Shoot regeneration via direct organogenesis from in vitro derived leaves of mulberry using thidiazuron and 6-benzylaminopurine

A brief culture of mulberry leaves for 8–10 days on MS medium with 18.17 μM TDZ followed by transfer to 8.88 μM BAP supplemented medium triggered high frequency shoot organogenesis (77.6–89.2%) and favoured shoot elongation in Morus spp. Shoot proliferation was highest in the presence of 2.22 μM BAP with induction of 9.4–10.6 shoots per culture. High frequency of root induction (76.0–86.6%) was observed on medium supplemented with 0.49 μM IBA whereas increase in the level of IBA (4.92 μM) resulted in induction of roots along with development of callus from the base of the shoots. The regenerated plants established in soil at higher frequency in rainy season compared to winter and summer.     

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.     

Stockplant factors affecting root initiation in cuttings of Triplochiton scleroxylon K. Schum., an indigenous hardwood of west africa

The ability of auxin-treated T. scleroxylon cuttings to root was affected by the prior management of potted stockplants. In undecapitated single-stem stockplants more cuttings from upper rather than lower mainstem nodes rooted; a difference paralleled by leaf water potential immediately after severance, although there was also a positive relationship with internode length. The rooting percentage of mainstem cuttings from unpruned stockplants ranged from 15% to 43% whereas that of cuttings from the lateral shoots of pruned stockplants ranged from 40% to 83%. Considerably more cuttings rooted from stockplants which were severely pruned than from those where decapitation removed only the top node; there seemed to be an inverse relationship with the number of shoots per plant and the carbohydrate: nitrogen ratio. However, in tall pruned stock- plants, more cuttings from lower lateral (basal) than from upper (apical) shoots rooted, although the differences between cuttings from basal and apical lateral shoots were less when the stockplants’ mainstems were orientated at 45° or kept horizontal, instead of vertically. Adding NPK 16 weeks before harvesting cuttings from 10-node vertical stockplants increased the rooting ability of cuttings from basal shoots without affecting the rooting of those from apical shoots. More lateral shoot cuttings rooted when two, instead of one or four lateral shoots were allowed to develop per stockplant, this being associated with less cutting mortality than occurred in pruned stockplants. In stock- plants with two shoots, cuttings from basal lateral shoots rooted better than those from apical shoots, although without competition from basal shoots. The rooting of apical shoots was enhanced by application of a complete fertilizer. The presence of basal shoots reduced the rooting ability of apical shoots even with the fertilizer application. Many of the effects of lateral branch position on rooting may be related to light intensity, for greater rooting percentages occurred among cuttings from lower, more shaded, than from upper less shaded branches. This positional effect was eliminated when branches were uniformly illuminated.     

Piriformospora indica promotes adventitious root formation in cuttings

Adventitious root formation in excised plant shoots is a crucial process in the vegetative propagation of many plant species, and insufficient rooting causes substantial losses in the propagation industry. Based on the various physiological effects on whole plants described for the basidiomycete Piriformospora indica, it was hypothesized that inoculation of the substrate with this endophyte should promote the generation and growth of adventitious roots in cuttings. Inoculation experiments were conducted to study the effects of P. indica on adventitious rooting in three plant species. Inoculation with P. indica dramatically enhanced the number and length of the adventitious roots in pelargonium and poinsettia. Root colonization parameters suggest that the interaction between the endophyte and cuttings had already occurred before physical contact. In contrast, petunia showed no rooting response to P. indica inoculation. Very fast root formation in this plant indicates that a minimum time period for the fungus–plant interaction is required for establishment of a promoting effect. P. indica-based biotechnology is proposed as a new tool for improving plant propagation systems of plant species or cultivars with low to moderate capacity of adventitious root formation.     

Daylength and photon flux density influence the growth regulator effects on morphogenesis in epicotyl segments of Troyer citrange

The optimal growth regulator addenda for adventitious shoot regeneration in epicotyl cuttings of Troyer citrange (Citrus sinensis×Poncirus trifoliata) varied with the conditions of illumination. The response to illumination and to growth regulators differed for the direct and the indirect (through callusing) pathways of regeneration. Shoot formation through the direct organogenic pathway decreased as the concentration of benzyladenine (BA) in the medium was increased in the range 2.2–22 μM, when the explants were incubated in the dark or under an 8 h daylength. For explants incubated under a 16 h daylength, the number of shoots formed increased with BA concentration. Optimal conditions of incubation for shoot formation through the direct pathway were either an 8 h daylength with a photon flux density of 74 μmol m⁻² s⁻¹ in the presence of 2.2 μM BA, or a 16 h daylength with a photon flux density of 37 μmol m⁻² s⁻¹ in the presence of 22 μM BA. Irrespective of the conditions of incubation, shoot formation through the indirect organogenic pathway was suppressed by the addition of 22 μM BA to the medium. Optimal conditions for shoot formation through this pathway were incubation under an 8 h daylength at a photon flux density of 74 μmol m⁻² s⁻¹ in the presence of 2.2 μM BA. At the optimal conditions indicated, the addition of the synthetic auxin naphtaleneacetic acid (0.54 μM) reduced shoot formation. Irrespective of the pathway of regeneration, the number of shoots formed decreased markedly with the distance of the cutting from the cotyledonary node.     

Inhibitory effects of riboflavin (Vitamin B2) on the in vitro rooting and nutrient concentration of explants of peach rootstock GF 677 (Prunus amygdalus × P. persica)

The effect of vitamin riboflavin (B₂) on in vitro rooting of the almond × peach hybrid clone GF 667 was studied. Riboflavin was added in five concentrations: 0 (control), 0.5, 1.0, 1.5 and 2.0 mg l⁻¹. After 4 weeks in culture, riboflavin did not stimulate adventitious rooting of the explants and rooting was very low in comparison to the control treatment. A high percentage of shoots on the rooting media containing the two highest concentrations of Vitamin B₂ had shown symptoms of chlorosis and apex necrosis.     

A liquid 2,4-D pulse increased shoot and root regeneration from leaf explants of adult Prunus rootstocks

Regeneration of adult plant material is one of the main limitations for successful Prunus rootstock transformation. Results herein show that a liquid pulse (90 min) of 2,4-dichlorophenoxyacetic acid (2,4-D) (1.7 μM), applied to leaf explants, greatly improved shoot regeneration in Marianna 2624 (Prunus cerasifera × munsoniana) and Myrobalan 605 AD (P. cerasifera); and induced roots in Adafuel (Prunus × amigdalo-persica) when placed in regeneration medium. Whole leaves and basal leaf explants of Marianna 2624 regenerated shoots in a higher proportion of explants after the pulse (up to 58.9% in whole leaves) than medium or tip leaf segments, whereas the leaf tip was the explant that showed less regeneration. In the regeneration medium, in the presence of BAP, NAA was more effective than 2,4-D. The application of an auxin pulse is a simple method that could enhance adult plant regeneration in commercial rootstocks.     

Differentiation of the dynamic variables affecting rooting ability in juvenile and mature cuttings of cherry (Prunus avium)

Summary

The rooting potential of four types or origins of Prunus avium cuttings from the same mature trees (over 20 years-old) was compared using a mist propagation bed during early Summer (June). The cuttings originated from juvenile sucker shoots of the current and previous year, and mature crown shoots (current year’s lateral ‘long-shoots’ and multi-year terminal ‘short shoots’). The morphological differences in inter-node length, stem diameter and leaf area between the four cutting types were highly significant (P = 0.05), leading to large differences in cutting volume and, so it is argued, to assimilate reserves. Juvenile cuttings rooted well (65% and 77% rooting for hardwood and softwood shoots, respectively), while mature cuttings rooted poorly (4% and 7% for mature hardwood and softwood cuttings, respectively). Leaf abscission was significantly more frequent in mature hardwood cuttings (16 – 78%) than in the other cutting types (1.6 – 9%) at the end of the propagation period. Leaf loss resulted from two processes: abscission and leaf rotting. Physiologically (i.e., in carbon assimilation, leaf transpiration and stomatal conductance), the four cutting types were not significantly different early in the post-severance period (day-4); but, by day-22, stomatal conductance was lowest in mature hardwood cuttings that still had leaves. At this time, the most physiologically-active unrooted cuttings were from juvenile hardwood and mature softwood shoots. The extent of physiological and morphological variability between cutting types and their probable impact on processes affecting rooting ability is complex and highly interactive. Consequently, it is not possible to explain the causes of the variation in rooting ability between juvenile and mature cuttings, although this study suggests that the constraints to rooting are likely to reflect physiological differences between the different cutting types. It is concluded that, to resolve the debate about factors that affect the rooting ability of juvenile and mature cuttings (i.e., ontogenetic vs. physiological ageing), there is a need to achieve morphological and physiological comparability in the tissues.     

Poststorage quality and rooting ability of Epipremnum pinnatum cuttings after treatment with ethylene action inhibitors

Summary

The influence of the ethylene action inhibitors STS and 1-MCP on poststorage performance and subsequent rooting of cuttings was investigated in Epipremnum pinnatum. Unfavourable storage conditions resulted in decreasing poststorage quality of single-eye cuttings, expressed as leaf drop and yellowing. 1-MCP and STS prohibited leaf drop and yellowing in E. pinnatum. STS pretreatment decreased rooting ability significantly, measured as percentage of rooted cuttings, number of roots, total root length and dry weight. STS caused severe injuries to the cuttings, which were worse if the cuttings were stored rather than propagated immediately. There was no significant difference in rooting between 1-MCP treated cuttings and untreated control in either stored or unstored cuttings. The influence of ethylene action inhibitors on rooting and a possible use of 1-MCP for practical use to increase poststorage performance in cuttings are discussed.     

三种生长调节剂对金边虎尾兰、月季与天竺葵扦插生根的影响

以金边虎尾兰、月季与天竺葵为试材,研究不同生长调节剂及其浓度对3种园林植物扦插生根的影响。结果表明:金边虎尾兰叶基段以100mg/L浓度的IBA或NAA扦插效果较好,叶尖段以IBA 100～200mg/L或NAA 200mg/L扦插效果较好,生根率均达到100%;100～200mg/L浓度的IBA与100mg/L浓度的NAA对月季茎插生根效果较好,生根率达75.6%～77.8%;天竺葵茎插各处理的生根率均达到100%,但从平均生根数上比较,100mg/L NAA或25mg/L IBA对其扦插生根效果最佳,25～300mg/L的IAA对其生根数有抑制作用。     

Contributions of the leaves and axillary shoots to rooting in Eucalyptus granáis Hill ex Maid,stem cuttings

In one-node stem cuttings of Eucalyptus granáis the absence of leaves was fatal. The optimum leaf area per leaf for rooting was 2-5 cm2, and in this range rooting was higher in one-leaf than in two-leaf cuttings. Axillary shoots started to develop soon after the cuttings were set and their growth was promoted by the presence of the subtending leaf. Thus, in one-leaf cuttings the shoot in the leafy axil grew, at the expense of the shoot in the defoliated axil, while in two-leaf cuttings the shoot in the upper (sub-opposite) axil tended to grow more quickly. Axillary shoots were also vigorous in cuttings from decapitated and disbudded mother plants. Rapid shoot growth was associated with low rates of abscission of the subtending leaf and relatively high rooting ability. Removing axillary shoots from cuttings accelerated leaf abscission and reduced rooting. The stem of the cutting often swelled and discoloured from the base, apparently due to the demand of the cutting for water. Additional leaf wounds had no effect on rooting (but increased mortality), while immersing cuttings in water before setting, or leaving them in a loose heap on the propagation bench, promoted rooting.