Influence of ventilation and sucrose on growth and leaf anatomy of micropropagated potato plantlets

Potato single nodes were cultured in vessels containing MS medium supplemented with 10, 20 and 30 g/l of sucrose. Vessels were closed with a clear polypropylene lid with or without 10 mm microporous polypropylene membrane. Sucrose concentration significantly increased plantlet height, shoot fresh weight and chlorophyll a content. Plantlets grown in ventilated vessels were significantly shorter, had lower shoot fresh weight and higher shoot dry weight than those in non-ventilated vessels. The highest leaf chlorophyll a content (21.83 mg/g fresh weight) was found in plantlets grown in ventilated vessels using MS medium with 20 g/l of sucrose, whereas those grown on medium with 10 g/l of sucrose had the highest chlorophyll b content (24.00 mg/g fresh weight). Total chlorophyll content was significantly higher when plantlets were grown in ventilated vessels containing medium with 10 or 30 g/l sucrose than in non-ventilated vessels. There was no significant difference in total chlorophyll content among plantlets grown in ventilated vessels with different concentrations of sucrose. Stomatal density was significantly lower when plants were grown under ventilated conditions. Leaf replica examination showed that stomata under non-ventilated condition were spherical with wide openings whereas, those in ventilated vessels were elliptical with narrow openings. Plantlets grown in non-ventilated vessels had thinner leaves and failed to build up a distinct defined upper epidermis, palisade parenchyma layer and spongy cells. On the other hand, leaves under ventilated conditions showed comparatively well organized layers with small intercellular space. The vascular system of leaves under the ventilated conditions demonstrated very well developed xylem unlike leaves under non-ventilated conditions. Thus, ventilated vessels with the 20 g/l of sucrose under ambient CO₂ in the growth room could successfully promote photomixotrophic culture and produce healthy plantlets.     

Callus induction and plant regeneration from embryonic axes of Kosteletzkya virginica

Kosteletzkya virginica, a perennial dicot halophytic species of the Malvaceae, is native to American salt marsh. It was introduced into China as a potential species to improve coastal wetlands and to develop ecologically sound saline agriculture. K. virginica adapts excellently to the tidal-flat habitats in China's east coast, with multiple eco-benefits; in particular, its seed oil could be used to produce biodiesel. The purpose of this study was thus to develop a standardized protocol to induce a high frequency of callus and subsequent plantlet regeneration system for a K. virginica breeding program with the final objective of applying transgenic techniques to improve seed oil yield. The embryonic axes of K. virginica were used as explants for callus induction, shoot induction from the callus and then adventitious root induction from the shoots on nine culture media with different hormone combinations. The best results were achieved on the following media: (1) 93.94% callus induction on MS medium supplemented with 1.0 mg L⁻¹ indole-3-acetic acid (IAA), 0.3 mg L⁻¹ kinetin, 30 g L⁻¹ sucrose and 8 g L⁻¹ agar; (2) 65.83% shoot induction on 1/2MS medium supplemented with 0.1 mg L⁻¹ IAA, 0.5 mg L⁻¹ zeatin, 30 g L⁻¹ sucrose and 8 g L⁻¹ agar; (3) 96.67% rooting on MS medium containing 30 g L⁻¹ sucrose and 8 g L⁻¹ agar. The survival rate of plantlets by organogenic regeneration was 85% after being transplanted into potting soil in flowerpots and placed in the greenhouse. This experiment indicates that we established successful callus induction and plant regeneration protocols for K. virginica.     

Nitrogen content determines adventitious rooting in Euphorbia pulcherrima under adequate light independently of pre-rooting carbohydrate depletion of cuttings

Root regeneration in shoot tip cuttings responds to graduated nitrogen (N) fertilization of stock plants. In pelargonium cuttings, reduced carbohydrate reserves caused by high N absorption by the donor plants and post-harvest storage of cuttings limit adventitious root formation, especially in low-light environments. In contrast, in chrysanthemum, similar carbohydrate reserves do not have this dominant effect on rooting capacity. The positive correlation between rooting capacity and internal N status is stable across a wide range of environments and is genotypically consistent for this species. However, the influence of N and carbohydrates on adventitious rooting of Euphorbia pulcherrima is unknown. We investigated the consequences of different N fertilization regimens applied to E. pulcherrima stock plants and cold and dark storage of the cuttings on N absorption, carbohydrate distribution, and rooting capacity of cuttings. Increasing time of stock plant cultivation with graduated N nutrition produced cuttings with N contents, ranging from 19 to 51 mg N g⁻¹ dry mass (DM). High N absorption resulted in low carbohydrate concentrations in cuttings, and subsequent storage decreased carbohydrate concentrations further, particularly in stems. Lower sucrose contents in leaves were correlated with reduced rooting of stored cuttings at a particular harvest date. However, despite the lower carbohydrate levels, root numbers and lengths correlated positively with internal N concentrations. These relationships remained stable in unstored and stored cuttings, even when overall rooting intensity was reduced under lower natural light during autumn. Multivariate regressions accounting for nitrogen content, sucrose content and daily light integral during rooting highlighted these relationships and explained up to 79% of rooting variances. We conclude N nutrition of stock plants and N absorption by cuttings are the dominant factors determining the rooting capacity of poinsettia when rooting occurs under sufficient light, as is commonly available during propagation. To maximize rooting capacity of poinsettia cuttings their nitrogen content should exceed a threshold of 40 mg N g⁻¹ DM.     

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.     

Influence of medium composition and vessel ventilation on in vitro propagation of Phillyrea latifolia L.

Micropropagation of Phillyrea latifolia L. a wild species present in Mediterranean coastal areas having drought and salt tolerance was performed using explants from adult plants. Shoots were induced from nodal explants on the Rugini’s initial medium (IM). Then these were proliferated on either Rugini olive medium (OM) or Linsmaier and Skoog (LS) medium, each supplemented with 2.22 μM 6-benzylaminopurine (BA) or 4.56 μM zeatin (Z). Rooting (66.1±11%) was induced on shoots grown in perlite soaked with half-strength Rugini olive proliferation medium (OMr) containing 2.69 μM α-naphthaleneacetic acid (NAA) and 160 mg l⁻¹ putrescine. Both shoot multiplication and rooting were performed using Magenta^® GA-7 (Sigma) vessels either non-permeable or permeable to gas exchanges. Contamination (about 40%) was observed during the first five passages notwithstanding the addition of cefotaxime to the culture medium, but a high proliferation rate (90%) of explants provided enough healthy plant material. The highest shoot proliferation was observed on LS medium and zeatin whereas the presence of the ventilated filters reduced fresh weight of explants growing on LS media and did not affect shoot growth on OM media. During rooting, the use of ventilated vessels in comparison with the closed ones enhanced development of roots, and doubled the dry weight of plantlets. The vessel ventilation combined with the artificial substrate (perlite) was beneficial for in vitro acclimatization of rooted Phillyrea plantlets.     

High frequency plant regeneration through adventitious multiple shoot organogenesis in epicotyl explants of Indian gooseberry (Emblica officinalis Gaertn)

Shoots regenerated adventitiously on epicotyl segments from in vitro seedlings of Emblica officinalis var. ‘Kanchan’. Epicotyls derived from 2-week-old aseptic seedlings were most responsive and produced a maximum number of 303 shoots per explant in Murashige and Skoog (1962) medium (MS) augmented with 8.8 μM N⁶-benzyladenine (BA) + 1.425 μM indole-3-acetic acid (IAA). Shoots readily elongated in MS lacking growth regulators and rooted in half-salt-strength MS (1/2 MS) supplemented with indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA). The highest rooting response was recorded in 1/2 MS containing 14.7 μM IBA. Plantlets were acclimatized inside the green house and 80% of the plantlets survived on transfer to garden soil.     

Light quality affects in vitro adventitious rooting and ex vitro performance of cherry rootstock Colt

The effect of light quality on in vitro rooting of cherry rootstock “Colt” was studied to evaluate adventitious root development, ex vitro acclimatization and plant growth performance. Adventitious rooting was dependent on the light quality to which the microcutting were exposed. Highest number of roots per microcuttings was recorded under dichromatic light (blue + red). This response could be ascribed to the control of a strong synergistic interaction between phytochromes and blue light photoreceptors. Red light was more effective on root elongation than blue light. In this response a strong synergistic interaction between phytochromes and blue light photoreceptors was suggested. The effect of light quality on the number of root/explant affected the plant during acclimation, scoring the highest level of plant survival in the blue-, red- and blue + red-exposed plantlets. The light quality effect was also observed under greenhouse culture conditions, as shown by growth parameters at the end of six months in plants growth. No specific role was observed for the photoequilibrium of phytochrome. The results reported in this work show that plantlets exposed to different light quality, during the in vitro rooting phase, retain the light quality effects in the subsequent plant acclimatization and greenhouse plant growth phase.     

Ethylene influences development and flowering of Ptilotus spp. in vitro and ex vitro

The genus Ptilotus has immense potential for ornamental horticulture but its commercial development has been hindered by propagation limitations. Poor seed quality and germination are reported. Cutting propagation is limited by cutting supply as the juvenile phase of Ptilotus is short. Micropropagation has been used in an attempt to overcome these difficulties but explants become floral in vitro and this causes plantlets to elongate. Ethephon has been used to control flowering of stock plants of many ornamental species. This study investigated the effect of ethephon applied to young (3-week-old, deflasked from tissue culture) and mature (1-year-old) Ptilotus plants in a greenhouse. A system of applying gaseous ethylene at 0, 100, 200 and 300 mg l⁻¹ to the headspace of in vitro plantlets in glass jars was developed and the response of in vitro plantlets to ethylene studied. One-year-old Ptilotus plants were treated with 500 mg l⁻¹ ethephon 2 days before pruning or 1 or 2 weeks after pruning. Ethephon application 2 days before pruning decreased the number of inflorescences and increased the number of shoots (compared to the control) but was phytotoxic. Ethephon applications of 150 or 300 mg l⁻¹ applied weekly or fortnightly to 3-week-old plants deflasked from tissue culture reduced plant height and number of inflorescences and at low concentrations increased the number of new shoots. A fortnightly application at 150 mg l⁻¹ is recommended. Previous reports on the effects of ethylene on inflorescence production on plantlets in vitro are limited. Our study showed that exposure of in vitro plantlets of P. nobilis to ethylene gas at 100 mg l⁻¹ for 1 h significantly increased the number of shoots and plant height but this did not occur for plantlets of P. spicatus. Plantlets of P. spicatus exposed to transient ethylene at 200 and 300 mg l⁻¹ showed significantly greater rooting (52.4%) than the control (13.6%).     

Encapsulation of shoot tips of guava (Psidium guajava L.) for short-term storage and germplasm exchange

Shoot tips obtained from in vitro grown plantlets of guava (Psidium guajava L.) were encapsulated in calcium alginate beads for short-term storage and germplasm exchange. A gelling matrix of 3% sodium alginate and 100 mM calcium chloride was found most suitable for formation of ideal calcium alginate beads. Maximum percent response for conversion of encapsulated shoot tips into plantlets was obtained on growth regulator free full strength liquid MS medium. The regrowth ability of encapsulated shoot tips was affected by medium strength and sucrose concentrations in the medium. Encapsulated shoot tips could be stored at low temperature (4 °C) up to 30 days with a survival frequency of 25%. After 60 days of storage under minimal growth conditions (sucrose lacking medium), about 75% encapsulated shoot tips were converted into plantlets when subcultured on 3% sucrose containing medium. Plantlets regenerated from encapsulated shoot tips were acclimatized successfully.     

Effect of position and orientation of leaflet explants with respect to plant growth regulators on micropropagation of Zamioculcas zamiifolia Engl. (ZZ)

Micropropagation studies on Zamioculcas zamiifolia Engl. (ZZ) as to the position and orientation of leaflet explants and plant growth regulators were carried out. Explants consisted of leaflet lamina from the basal or apical part of the leaflet with or without petiolule or midrib that were placed vertically into the medium except for the explants with midrib from the basal part of the leaflet that were placed horizontally as well. The explants were cultured on solid Murashige and Skoog medium (MS) with 30 g l⁻¹ sucrose, supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 2 or 4 mg l⁻¹ and 6-benzyladenine (BA) at 0 or 4.44 μM in all (four) possible combinations, or with 1-naphteleneacetic acid (NAA) at 0 or 5.38 μM and BA at 0 or 4.44 μM in all (four) possible combinations (establishment medium). The morphogenic response was direct from all types of leaflet explants and varied only with respect to different plant growth regulators of the medium: 2,4-D combined or not with BA formed somatic embryo-like structures; NAA alone produced tubers and roots; BA alone resulted mainly in leaves; NAA combined with BA produced mainly roots. The intensity of the response varied accordingly to the explant type and orientation. Explants with petiolule or midrib from the basal part of the leaflet showed the highest morphogenic response compared to explants without petiolule or midrib or to explants from the apical part of the leaflet, in all the plant growth regulator combinations used. Explants with midrib from the basal part of the leaflet placed vertically into the media showed higher morphogenic response compared to those placed horizontally on the medium surface. With the objective to regenerate plantlets, explants were subcultured on MS with NAA and BA at various concentrations based on the explant response in the establishment medium, taking into consideration the initial explant type. The initial explant type did not affect the response in the subculture. Most plantlets (a tuber with roots and one leaf with one pair of leaflets) were produced by explants with embryo-like structures induced in a medium with only 2,4-D. Explants with tubers induced in a medium with NAA gave plantlets at 65–85% when subcultured in a medium with 4.44 μM BA alone or in combination with 2.69 μM NAA. Explants with leaves induced in a medium with BA and explants with roots induced in a medium with NAA and BA gave plantlets at low percentages (20–40%). The best response was produced by explants with embryo like structures induced in a medium with only 2,4-D which gave plantlets at 100% when subcultured in the medium with 2.69 μM NAA and 2.22 μM BA. Plantlets raised in different treatments were transplanted ex vitro after 22 weeks and exhibited 80–100% survival.     

In vitro germination of walnut (Juglans regia L.) embryos

The present studies were undertaken with a view to standardize the medium and culture conditions for embryo culture of five cultivars of walnut viz., ACO 38853, Netar Akhrot, Gobind, Solding Selection and Blackmore. Embryos from mature fruits were aseptically excised and cultured on MS medium supplemented with different combinations of BAP, kinetin and GA₃. Best performing medium was MS with 0.5 mg l⁻¹ kinetin, 0.5 mg l⁻¹ BAP and 2 mg l⁻¹ GA₃ yielding 66.6% germination in Netar Akhrot after 12 days of culturing. Percent germination of excised embryos was higher when GA₃ and cold treatments were simultaneously applied as compared to those when applied separately. Netar Akhrot was found to be the best responding cultivar, which had a range of 25–66.6% embryo germination under different culture conditions. Plantlets with shoots and roots have been obtained in Netar Akhrot and ACO38853 and are transferred to soil after hardening.     

Development of a plant regeneration system from seed-derived calluses of centipedegrass [Eremochloa ophiuroides (Munro.) Hack]

The objective of this study is to establish plant regeneration system with the seed of the new Chinese selection “E-126”of centipedegrass [Eremochloa ophiuroides (Munro.) Hack] as explant. In present study, the following results were obtained: (1) The medium formulation most suitable for calluses induction was identified to be MS with 1.0 mg l⁻¹ 2,4-D + 30 g l⁻¹ mannitol + 50 ml l⁻¹ coconut milk and the ratio of calluses induction was 96.0%, including 5.2% of yellow granule calluses induction. The above medium formulation was adopted for subculture. (2) The rate of shoot regeneration from yellow granular calluses was 98.0% by MS optimum medium formulation with 2.0 mg l⁻¹ KT + 50 ml l⁻¹ coconut milk. The differentiated rate retained as high as 88.0% even after 5 times of subculture and 18.6% after 15 times of subculture. The optimum medium formulation for shoot growth was identified to be MS medium plus 2.0 mg l⁻¹ BAP, 0.8 mg l⁻¹ NAA and 50 ml l⁻¹ coconut milk. (3) The optimum medium for shoot rooting was identified to be MS medium with 0.6 mg l⁻¹ NAA + 50 ml l⁻¹ coconut milk, and the rooting rate to be 98.0%. The survival rate of transplanted plantlets from tubes to basin with soil was 92.0%. In conclusion, the plant regeneration system was successfully developed in this study, which may provide basic reference for screening of somaclonal variants and genetic transformation of centipedegrass.     

苗龄与基质对黄瓜转基因组培苗驯化成活率的影响

以黄瓜转基因组培苗为材料,研究了不同苗龄、栽培基质和移栽前培养基中的蔗糖浓度对组培苗根系发育及驯化成活率的影响。结果表明：将转基因组培苗置于蔗糖浓度为6 %的1/2 MS培养基中生长28 d,随后移至蔗糖浓度为3 %的培养基中培养7 d,在组培苗六叶一心时,移栽于草炭∶珍珠岩∶河沙=1 V ∶1 V ∶1 V的基质中驯化,最有利于组培苗根系的发育,驯化成活率可达98.7 %,且经驯化成活的植株均能正常开花结果。     

In vitro shoot proliferation and enhancement of rooting for the large-scale propagation of yellow bamboo (Bambusa vulgaris ‘Striata’)

Continuous and rapidly proliferating axillary shoots were raised from axillary buds in secondary branches of adult field culms and nursery grown 1-year-old tissue culture-raised plants of Bambusa vulgaris ‘Striata’. Shoots continuously proliferated in a MS medium containing 4 mg L⁻¹ 6-benzyladenine (BA). The effects of indole butyric acid (IBA) levels, a pretreatment with thidiazuron (TDZ) (1-phenyl-1-([1,2,3-thidiazol-5-yl])urea) and illumination on rooting, were investigated after 6 months of shoot proliferation. A rooting medium with IBA at 3 mg L⁻¹ was optimum for root induction. Shoots of adult field culms that were proliferated in the presence of BA when induced to root in this medium resulted in 40% rooting in 27 days. In vitro shoots raised from 1-year-old tissue cultured plants showed 92% rooting under the same conditions. Rooting was enhanced when the relatively difficult-to-root in vitro shoots from adult field culms were pretreated with 0.5 mg L⁻¹ TDZ for two to three subcultures before placing in the root induction medium. Continuously illuminated shoots pretreated with TDZ for three subcultures showed 100% rooting compared to 83% rooting of shoots that were exposed to a 12 h photoperiod. These findings have been applied in the large-scale propagation of this species.     

Rooting response of shoot cuttings from three peach growth habits

Current year shoot cuttings were collected in October and August from three growth habits of peach (Compact, Pillar, and Standard) and treated with one of four concentrations of indole butyric acid (0, 250, 1250, and 2500 mg L⁻¹ IBA). Rooting response was measured after 5 weeks in the greenhouse. Little or no rooting occurred with cuttings from any growth habit that was collected in October or in August when treated with 0 and 2500 mg L⁻¹ IBA. In August, the number of shoots that rooted was greater in cuttings from Pillar (79 and 45%) than Compact (13 and 3%) treated with 250 and 1250 mg L⁻¹ IBA, respectively. Cuttings from Standard trees had intermediate rooting of 56 and 6% at 250 and 1250 mg L⁻¹ IBA, respectively. Pillar trees consistently grew more roots with greater root length per cutting than the other growth habits. It is proposed that differences in rooting response among the growth habits may be associated with differences in endogenous auxin concentration that had been found in previous studies. Within peach and possibly other fruit trees, the capacity of shoot cuttings to develop adventitious roots can vary by cultivar and successful root induction with exogenous plant growth regulators may depend, in part, on endogenous hormone levels.     

Effect of auxin treatments,cuttings’ collection date and initial characteristics on Paeonia ‘Yang Fei Chu Yu’ cutting propagation

Paeonia ‘Yang Fei Chu Yu’ is one of the most popular and commercially valuable cultivars of herbaceous peony. The study was performed to explore propagation techniques by cuttings for the nursery industry. Results showed that the stem cuttings pretreated with 2000 mg L⁻¹ indole-3-butyric acid (IBA) in quick-dip method got the best rooting traits (rooting percentage is 86.7%, root number is 23.1 and root length per rooted cutting is 6.4 cm). Therefore pre-treatment with 2000 mg L⁻¹ IBA is recommended.     

Seed inoculation with Azospirillum mitigates NaCl effects on lettuce

Data on the growth-promoting effects of Azospirillum on lettuce exposed to either normal or saline conditions, is scarce. Lactuca sativa L., cv Mantecosa seeds were colonized with A. brasilense Sp245 cells during imbibition. Germination percentages were determined after 7 d treatments with 0, 30, 50 or 80 mol m⁻³ NaCl. In another experiment, seeds germinated in Hoagland were irrigated for 30 d with 0, 30, 50 or 80 mol m⁻³ NaCl supplemented media. Vegetative growth proceeded in a growth chamber with a 13–11 h day–night cycle. Buffer-imbibed seeds were considered non-inoculated controls. Plant samples were taken at 0, 14, 20, and 30 d after the onset of NaCl treatments and dissected in aerial and root portions. The weights of both tissues were measured. Azospirillum-inoculated seeds had significantly higher germination percentages than controls in all treatments. Inoculated dried seeds stored up to 30 d maintained such characteristic in most of the treatments, particularly at 80 mol m⁻³ NaCl. Plants grown from inoculated seeds and irrigated with saline media displayed higher total fresh and dry weights and biomass partition to the aerial portion, than non-inoculated controls. Azospirillum-inoculated lettuce seeds had better germination and vegetative growth than non-inoculated controls after being exposed to NaCl.     

Production of protocorm-like bodies with bioreactor and regeneration in vitro of Oncidium ‘Sugar Sweet’

To produce mass propagules of Oncidium ‘Sugar Sweet’, we tested the feasibility of producing protocorm-like bodies (PLBs) using 5 l balloon type air-lift bioreactors, and selected an optimal culture medium for shooting and rooting in vitro. The results showed that liquid bioreactor cultures were more efficient for PLBs proliferation when compared to solid- and liquid-agitated flask cultures. The maximum PLBs biomass (326.3 g per bioreactor, FW) was obtained in the immersion bioreactor culture, with the growth ratio reaching 10.9 after 50 days of culture. This was obviously higher than the ebb and flood bioreactor culture. During bioreactor culture, sucrose and electric conductivity (EC) in the culture medium were negatively correlated with PLBs growth; the highest PLBs fresh and dry biomass was obtained 40 days after culture. An inoculation density of 20 g (FW) was optimal for PLBs growth in a 3 l working volume of 5 l bioreactor. Furthermore, MS medium supplemented with 2.0 mg l⁻¹ BA for shooting and 0.5 mg l⁻¹ IBA for rooting was optimal during in vitro culture, the plantlets were successfully established in the potting substrate.     

Culture method and photosynthetic photon flux affect photosynthesis,growth and survival of Limonium ‘Misty Blue’ in vitro

Microcuttings (shoots each with two leaves) of Limonium ‘Misty Blue’ were cultivated in vitro for 28 days under photoautotrophic (sucrose-free culture medium; CO₂ and photosynthetic photon flux (PPF) enriched conditions), photomixotrophic (medium with 30 g l⁻¹ sucrose; CO₂ and PPF enriched conditions) and heterotrophic (medium with 30 g l⁻¹ sucrose; CO₂ non-enriched conditions) methods. Several growth variables were measured during and at the end of cultivation: shoot fresh and dry weight, percentage of shoot dry matter, root fresh weight, number of leaves, leaf area, chlorophyll and sugar content of leaves, stomatal density and size, net photosynthetic rate (NPR) and percent survival of plantlets ex vitro. Plantlets grown in photoautotrophic and photomixotrophic methods had more leaves, high chlorophyll and sugar contents, high NPR, and showed high percent survival. However, these plantlets possessed less number of stomata per square millimeter. In contrast, the plantlets grown by the heterotrophic method showed decreased values of these growth variables except for the number of stomata per square millimeter. These results indicate that CO₂ enrichment for plantlets in vitro at a relatively high PPF would promote photosynthesis and hence growth of chlorophyllous explants/plantlets in vitro. The resulting plantlets were acclimatized better and sooner on ex vitro transplantation.     

Encapsulation for in vitro short-term storage and exchange of ginger (Zingiber officinale Rosc.) germplasm

Synseeds of ginger (Zingiber officinale) were produced using aseptically proliferated 2-week old encapsulating explants (microshoots) upon complexation of 4% sodium alginate prepared in Murashige and Skoog (1962) medium (MS) and 100 mM calcium chloride. Conversion of synseeds into plantlets (conversion) was recorded as 66% and 53% on MS (3% sucrose) and on MS (3% sucrose) + 2.5 mg/l BA media, respectively. However, shoots/synseed were significantly higher on MS (3% sucrose) + 2.5 mg/l BA medium. For short-term storage of germplasm, sucrose-dehydrated synseeds were found better than air-dehydrated or fresh synseeds. Synseeds dehydrated in 0.25 M sucrose liquid medium for 16 h and stored in cryovials (with out medium) at 25 °C for 8 weeks and 12 weeks exhibited 53% and 13% conversion, respectively, on MS (3% sucrose) + 2.5 mg/l BA medium. Plantlets obtained from stored synseeds were hardened, established successfully ex vitro and were morphologically similar to each other as well as their mother plants. This synseed protocol could be useful for short-term storage and exchange of germplasm of ginger between national as well as international laboratories.