Insight into generation of induced mesenchymal stem cells from induced pluripotent cells

Mesenchymal stem cells(MSCs)have the potential for use in cell-based regenerative therapies.Currently,hundreds of clinical trials are using MSCs for the treatment of various diseases.However,MSCs are low in number in adult tissues;they show heterogeneity depending upon the cell source and exhibit limited proliferative potential and early senescence in in vitro cultures.These factors negatively impact the regenerative potential of MSCs and therefore restrict their use for clinical applications.As a result,novel methods to generate induced MSCs(iMSCs)from induced pluripotent stem cells have been explored.The development and optimization of protocols for generation of iMSCs from induced pluripotent stem cells is necessary to evaluate their regenerative potential in vivo and in vitro.In addition,it is important to compare iMSCs with primary MSCs(isolated from adult tissues)in terms of their safety and efficacy.Careful investigation of the properties of iMSCs in vitro and their long term behavior in animals is important for their translation from bench to bedside.     

Factors influencing pathogenicity of Fusarium tumidum on gorse (Ulex europaeus)

Factors promoting pathogenicity of Fusarium tumidum on gorse (Ulex europaeus) were determined to develop a novel strategy for delivering this potential mycoherbicide using insects as vectors of inoculum. Fusarium tumidum sprayed as a suspension of 1×10⁶ conidia mL^?1 on at least 50% of a gorse plant reduced shoot dry weight by 45% (P<0.05). A minimum of 910 viable conidia were required to cause a lesion on leaves. The leaves and flowers were more susceptible to infection than stems, spines and pods. Generally, wounding of gorse leaves and stem increased F. tumidum infection, most likely through releasing nutrients that enhanced conidial germination and hyphal growth. We showed in a separate experiment that conidial germination (93%) and germ tube length (407 µm) were greater when incubated in 0.2% gorse extract solution for 24 h than in water (62% germination, germ tube length 42 µm). Inoculation of gorse with a F. tumidum conidial suspension supplemented with 0.2% gorse extract resulted in a shoot dry weight reduction (P=0.012) equivalent to that of plants that were wounded and inoculated. It is concluded that wounding of older tissues (which mimics insect damage) is required to facilitate F. tumidum infection of mature gorse plants.     

江西常绿阔叶林木本植物不同冠层高度当年生小枝茎构型对叶生物量的影响

以江西阳际峰自然保区69种木本植物为对象,采用标准化主轴回归(SMA)的方法,对不同冠层高度当年生小枝的构件生物量(叶生物量、茎生物量、小枝生物量)与茎构型特征(茎直径、茎长度、茎宽长比、茎体积和茎密度)进行了分析,研究当年生小枝茎构型对叶生物量的影响.结果表明:不同冠层高度及生活型间叶生物量、茎生物量、小枝生物量、茎直径、茎长度、茎宽长比及茎体积均无显著差异,而茎密度差异显著.不同冠层高度及生活型间,叶生物量与茎生物量及总生物量之间均呈显著等速生长关系.小枝叶生物量分别与茎直径、茎体积呈显著异速生长关系,且该异速生长指数在不同冠层高度无显著差异.茎长度、茎宽长比及茎密度对当年生小枝叶生物量变异的解释力较小(<24%).冠层高度和生活型对小枝叶-茎生物量的等速分配关系影响不显著.小枝茎构型中,相比于茎长度、茎宽长比以及茎密度,茎的直径与体积对当年生小枝叶生物量的影响更大,冠层高度对当年生小枝茎构型与叶生物量间的异速分配模式无显著影响.     

Relationship of Auxin Transport to Branch Dimorphism in Cordyline, a Woody Monocotyledon

Lanolin containing ³H-IAA was applied to apical cut ends of stem segments and decapitated plants of Cordyline terminalis, a monocotyledon possessing true secondary growth. Distribution of extracted label showed that: 1) in horizontal stems up to 7 times as much label accumulates in the lower surface compared to the upper surface, and this ratio increases with distance from the cut apical surface; 2) in upright stems most label (62%) occurs in the cambial and cortical regions, external to the primary and secondary vascular tissues; and 3) in upright stems having the cambium and cortical regions removed by a girdle, label accumulates above the girdle (c. 10 times greater than the control). These data are in accord with a theory of branch dimorphism derived from morphological studies, which suggested that high auxin levels initiate and maintain rhizomes and low levels initiate leafy shoots.     

Release of cotyledonary shoots of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) seedlings from inhibition as related to endogenous zeatin and ethylene and exogenous IAA and benzyladenine

This paper deals with apical dominance using a dicotylar model obtained after decapitation of pea seedlings with two shoots — one dominant and the other inhibited. When the dominant shoot was decapitated the inhibited one is released from inhibition and after 24 to 72 h begins to grow. However, the levels of trans-zeatin and production of ethylene increase within 4 and 6 hours respectively after release from inhibition, and within an interval of 72 h the levels of both phytohormones begin gradually to decrease. This indicates that also in this model, the release from apical dominance is associated with an increase in the level of cytokinin zeatin and, thereafter, also with an increased production of ethylene. If indolyl-3-acetic acid (IAA) is applied on the decapitated main stem after decapitation of the dominant shoot, the growth of the initially inhibited one is very strongly retarded; if, however, IAA is applied on the decapitated dominant shoot, this inhibition is significantly weaker. This means that the inhibiting effect of IAA on the inhibited shoot originates to a greater degree from the main stem rather than from the dominant shoot. The effect of benzyladenine (BA) is transferred equally from the decapitated main stem and from the decapitated dominant shoot because the initially inhibited shoot begins to grow as well as also other shoots from serial cotyledonary buds.     

Application of mesenchymal stem cells derived from human pluripotent stem cells in regenerative medicine

Mesenchymal stem cells (MSCs) represent the most clinically used stem cells in regenerative medicine. However, due to the disadvantages with primary MSCs, such as limited cell proliferative capacity and rarity in the tissues leading to limited MSCs, gradual loss of differentiation during in vitro expansion reducing the efficacy of MSC application, and variation among donors increasing the uncertainty of MSC efficacy, the clinical application of MSCs has been greatly hampered. MSCs derived from human pluripotent stem cells (hPSC-MSCs) can circumvent these problems associated with primary MSCs. Due to the infinite self-renewal of hPSCs and their differentiation potential towards MSCs, hPSC-MSCs are emerging as an attractive alternative for regenerative medicine. This review summarizes the progress on derivation of MSCs from human pluripotent stem cells, disease modelling and drug screening using hPSC-MSCs, and various applications of hPSC-MSCs in regenerative medicine. In the end, the challenges and concerns with hPSC-MSC applications are also discussed.     

The influence of plant growth regulators on explant performance, bud break, and shoot growth from large stem segments of Acer saccharinum L

Benzyladenine (BA) and/or gibberellic acid (GA₃) were applied in 20% white exterior latex paint separately at 0, 0.3, 1, 3, 10, or 30 mM; and at 1, 10, or 30 mM of each plant growth regulator (PGR) in a 3 × 3 factorial to 40 cm long stem segments of Acer saccharinum L. Softwood shoots were forced from these stem segments at various times of the year in a greenhouse and in a laboratory, these resulting shoots were surface disinfested and used as explants in vitro on Driver and Kuniyuki Walnut medium with 0 or 0.01 μM thidiazuron (TDZ). There was some response to the plant growth regulators applied in paint for shoot production from the stem segments and in vitro. Explants from softwood shoots forced from stems painted with 3 mM BA and cultured on medium with 0.01 μM TDZ produced more shoots than explants taken from softwood shoots forced with other BA concentrations or controls. Callus also grew significantly more on explants from stems treated with 3 mM BA cultured on 0.01 μM TDZ than explants harvested from stems painted with other concentrations of BA excluding 10 mM BA. When stem segments treated with BA plus GA₃ were compared as a group to controls, more and longer softwood shoots grew on the stems painted with PGRs when all four runs were pooled (Sept. 2005 through Feb. 2006). Application of PGRs in paint extends the season of production of softwood shoots that may be used as explant materials and their subsequent performance in vitro.     

Size structure of current-year shoots in mature crowns

Characteristics of current-year shoot populations were examined for three mature trees of each of three deciduous broad-leaved species. For first-order branches (branches emerging from the vertical trunk) of the trees examined, lengths or diameters of all current-year shoots were measured. Total leaf mass and total current-year stem mass of first-order branches were estimated using an allometric relationship between leaf or stem mass and length or diameter of current-year stems. For each tree, the number of current-year shoots on a first-order branch was proportional to the basal stem cross-sectional area of the branch. On the other hand, first-order branches had shoot populations with size structures similar to each other. As a result, the leaf mass of a first-order branch was proportional to the basal stem cross-sectional area of the branch, being compatible with the pipe-model relationship. All current-year shoot populations had positively skewed size structures. Because small shoots have a larger ratio of leaf mass to stem mass than large shoots, first-order branches had an extremely large ratio of leaf mass to current-year stem mass. This biased mass allocation will reduce costs for current stem production, respiration and future radial growth, and is beneficial to mature trees with a huge accumulation of non- photosynthetic organs. The allometric relationships between leaf mass and basal stem diameter and that between leaf mass and current-year stem mass of first-order branches were each similar across the trees examined. Characteristics of shoot populations tended to offset inter-species diversity of shoot allometry so that branch allometry shows inter-species convergence.     

Strategies to improve regenerative potential of mesenchymal stem cells

In the last few decades, stem cell-based therapies have gained attention worldwide for various diseases and disorders. Adult stem cells, particularly mesenchymal stem cells (MSCs), are preferred due to their significant regenerative potential in cellular therapies and are currently involved in hundreds of clinical trials. Although MSCs have high self-renewal as well as differentiation potential, such abilities are compromised with “advanced age” and “disease status” of the donor. Similarly, cell-based therapies require high cell number for clinical applications that often require in vitro expansion of cells. It is pertinent to note that aged individuals are the main segment of population for stem cell-based therapies, however; autologous use of stem cells for such patients (aged and diseased) does not seem to give optimal results due to their compromised potential. In vitro expansion to obtain large numbers of cells also negatively affects the regenerative potential of MSCs. It is therefore essential to improve the regenerative potential of stem cells compromised due to “in vitro expansion”, “donor age” and “donor disease status” for their successful autologous use. The current review has been organized to address the age and disease depleted function of resident adult stem cells, and the strategies to improve their potential. To combat the problem of decline in the regenerative potential of cells, this review focuses on the strategies that manipulate the cell environment such as hypoxia, heat shock, caloric restriction and preconditioning with different factors.     

Dormancy and spring development of lateral buds in mulberry (Morus alba)

Patterns of spring development of lateral buds of mulberry (Morus alba L. cv. Shin-ichinose) coppice shoots on 11-year-old low-pruned stumps varied in response to girdling, pruning and arching. The erect controls showed a weak acrotonic (apex-favoring) growth habit, in which the majority of the buds, including the basal ones, sprouted and elongated in mid- and late April, and hence there was a prolonged imposition of dominance on the upper laterals in mid- and late May. In contrast, early spring girdling or pruning enhanced the activity of the upper buds of the proximal (lower) halves of the girdled stems or of the pruned stems, resulting in considerable dominance of the laterals from such buds in late April. Arching markedly inhibited buds on the under side of the arched stems, leading to poor shoots. By late April, the buds on the adaxial (upper) side readily grew into new vertical shoots, which dominated over the lateral ones. When studied by a multiple-node-cutting test, increased length of segments of post-dormant mulberry stems was accompanied by decreased bud activity of the segments and by decreased breaking ability of the lower buds within the segments, suggesting the importance of roots in the weak acrotonic habit of the erect stem in spring. By contrast, the acropetal influences of the attached stems can in part affect dominance relationships, perhaps mediated through competition for factors translocated from the roots. Continuous basal applications of abscisic acid inhibited bud break and shoot growth of the postdormant stem segments, but these inhibitory effects could be reversed by applied gibberellic acid A₃ (GA₃). Two phases of lateral bud dormancy in erect mulberry coppice shoots were identified. The first was characterized by a smaller breaking capacity in the upper buds than in the lower ones and hence by a basitonic (base-favoring) gradient in bud growth potential. The second phase corresponded to a restoration of these capabilities in the upper buds and to a change towards a linear gradient in bud growth potential, with disappearance of the dormant condition, in February and March. This gradient change during dormancy release may represent the physiological basis for the weak acrotonic habit of erect mulberry stems in spring.     

黄土高原北部典型灌丛枝条生物量估算模型

于2015年8月末在陕西神木县六道沟小流域采集200个柠条和210个沙柳枝条,测定枝条的基径(D)、长度(H)、含水量(W₀)、鲜质量(W_F)和干质量(W),选用指数函数和异速生长方程建立了4种由枝条形态指标估算枝条生物量的简易模型,并对模型的拟合效果进行验证. 结果表明: 对于柠条和沙柳灌丛,基于D、H二者组合变量(D²H)的异速生长方程是估算枝条生物量的最优模型,该模型经线性转化后可以消除生物量数据的异方差性,且拟合效果最优,决定系数(R²)最大,平均误差(ME)、平均绝对误差(MAE)、总相对误差(TRE)、平均系统误差(MSE)和平均绝对百分误差(MPSE)整体上最小,基本满足生态学研究的精度要求.     

Auxin transport and the regulation of petiole abscission in cotton (Gossypium hirsutum L.): A comparison of indol-3yl-acetic acid and phenylacetic acid

Distal applications of indol-3yl-acetic acid (IAA) to debladed cotyledonary petioles of cotton (Gossypium hirsutum L.) seedlings greatly delayed petiole abscission, but similar applications of phenylacetic acid (PAA) slightly accelerated abscission compared with untreated controls. Both compounds prevented abscission for at least 91 h when applied directly to the abscission zone at the base of the petiole. The contrasting effects of distal IAA and PAA on abscission were correlated with their polar transport behaviour-[1-¹⁴C]IAA underwent typical polar (basipetal) transport through isolated 30 mm petiole segments, but only a weak diffusive movement of [1-¹⁴C]PAA occurred.Removal of the shoot tip substantially delayed abscission of subtending debladed cotyledonary petioles. The promotive effect of the shoot tip on petiole abscission could be replaced in decapitated shoots by applications of either IAA or PAA to the cut surface of the stem. Following the application of [1-¹⁴C]IAA or [1-¹⁴C]PAA to the cut surface of decapitated shoots, only IAA was transported basipetally through the stem. Proximal applications of either compound stimulated the acropetal transport of [¹⁴C]sucrose applied to a subtending intact cotyledonary leaf and caused label to accumulate at the shoot tip. However, PAA was considerably less active than IAA in this response.It is concluded that whilst the inhibition of petiole abscission by distal auxin is mediated by effects of auxin in cells of the abscission zone itself, the promotion of abscission by the shoot tip (or by proximal exogenous auxin) is a remote effect which does not require basipetal auxin transport to the abscission zone. Possible mechanisms to explain this indirect effect of proximal auxin on abscission are discussed.     

Stem biomechanics of three columnar cacti from the Sonoran Desert

The allometric relationship of stem length L with respect to mean stem diameter D was determined for 80 shoots of each of three columnar cactus species (Stenocereus thurberi, Lophocereus schottii, and S. gummosus) to determine whether this relationship accords with that predicted by each of three contending models purporting to describe the mechanical architecture of vertical shoots (i.e., geometric, stress, and elastic similitude, which predict L proportional to D(alpha), with alpha = 1/1, 1/2, and 2/3, respectively). In addition, anatomical, physical, and biomechanical stem properties were measured to determine how the stems of these three species maintain their elastic stability as they increase in size. Reduced major axis regression of L with respect to D showed that alpha = 2.82 ± 0.14 for S. thurberi, 2.32 ± 0.19 for L. schottii, and 4.21 ± 0.31 for S. gummosus. Thus, the scaling exponents for the allometry of L differed significantly from that predicted by each of the three biomechanical models. In contrast, these exponents were similar to that for the allometry previously reported for saguaro. Analyses of biomechanical data derived from bending tests performed on 30 stems selected from each of the three species indicated that the bulk stem tissue stiffness was roughly proportional to L2, while stem flexural rigidity (i.e., the ability to resist a bending force) scaled roughly as L3. Stem length was significantly and positively correlated with the volume fraction of wood, while regression analysis of the pooled data from the three species (i.e., 90 stems) indicated that bulk tissue stiffness scaled roughly as the 5/3-power of the volume fraction of wood in stems. These data were interpreted to indicate that wood served as the major stiffening agent in stems and that this tissue accumulates at a sufficient rate to afford unusually high scaling exponents tot stem length with respect to stem diameter (i.e., disproportionately large increments of stem length with respect to increments in stem diameter). Nevertheless, the safety factor against the elastic failure of stems (computed on the basis of the critical buckling height divided by actual stem length) decreased with increasing stem size tot each species, even though each species maintained an average safety factor equal to two. We speculate that the apparent upper limit to plant height calculated for each species may serve as a biomechanical mechanism for vegetative propagation and the establishment of dense plant colonies by means of extreme stem flexure and ultimate breakage, especially for S. gummosus.     

Micropropagation of <Emphasis Type="Italic">Theobroma cacao</Emphasis> L. using somatic embryo-derived plants

Summary A micropropagation protocol was developed using cacao somatic embryo-derived plant as a source for nodal and apical stem explants, and apical microcuttings. Microcuttings were efficiently rooted and developed into plantlets. Axillary meristems within the remaining decapitated plantlets subsequently developed and were used for production of additional microcuttings, with an average 2.4 growing shoots per decapitated stem. The remaining plantelts were maintained as microcutting stock plants. When nodal stem explants were cultured on thidiazuron medium, axillary buds proliferated and developed into shoots, which were excised and rooted. However, the efficiency of this method is lower than rooting of apical microcuttings harvested directly from stock plants. During root induction, short treatment with indole-3-butyric acid (IBA) increased the total percentage of rooted microcuttings up to 89%. Longer exposures to IBA increased the average number of roots per microcutting (from 1.7 to 5.2). Plant acclimatization after rooting was achieved with an average success of 87%. During several months of growth in the greenhouse, the micropropagated plants developed functional taproots. Currently, cocoa plants produced by this micropropagation method have been successfully acclimated to field conditions in Ivory Coast, Ghana, and Saint Lucia.     

Mesenchymal stromal cells from umbilical cord blood

Mesenchymal Stromal Cells (MSC) are key candidates for cellular therapies. Although most therapeutic applications have focused on adult bone marrow derived MSC, increasing evidence suggests that MSC are present within a wide range of tissues. Umbilical cord blood (CB) has been proven to be a valuable source of hematopoietic stem cells, but its therapeutic potential extends beyond the hematopoietic component suggesting regenerative potential in solid organs as well. There is evidence that other stem or progenitor populations, such as MSC, exist in CB which might be responsible for these effects. Many different stem and progenitor cell populations have been postulated with potential ranging from embryonic like to lineage-committed progenitor cells. Based on the confusing data, this review focuses on a human CB derived, plastic adherent fibroblastoid population expressing similar characteristics to bone marrow derived MSC. It concentrates especially on concepts of isolation and expansion, comparing the phenotype with bone marrow derived MSC, describing the differentiation capacity and finally in the last the therapeutic potential with regard to regenerative medicine, stromal support, immune modulation and gene therapy.     

Mass acquisition of human periodontal ligament stem cells

The periodontal ligament (PDL) is an essential fibrous tissue for tooth retention in the alveolar bone socket. PDL tissue further functions to cushion occlusal force, maintain alveolar bone height, allow orthodontic tooth movement, and connect tooth roots with bone. Severe periodontitis, deep caries, and trauma cause irreversible damage to this tissue, eventually leading to tooth loss through the destruction of tooth retention. Many patients suffer from these diseases worldwide, and its prevalence increases with age. To address this issue, regenerative medicine for damaged PDL tissue as well as the surrounding tissues has been extensively investigated regarding the potential and effectiveness of stem cells, scaffolds, and cytokines as well as their combined applications. In particular, PDL stem cells (PDLSCs) have been well studied. In this review, I discuss comprehensive studies on PDLSCs performed in vivo and contemporary reports focusing on the acquisition of large numbers of PDLSCs for therapeutic applications because of the very small number of PDLSCs available in vivo.     

Ethylene Evolution Changes in Tilted Fraxinus mandshurica Rupr. var. japonica Maxim. Seedlings in Relation to Tension Wood Formation

The effects of ethylene on tension wood formation were studied in 3-year-old Fraxinus mandshurica Rupr. var. japonica Maxim, seedlings in two separate experiments. In experiment 1, ethylene evolution of buds and stems was measured using gas chromatography after 0, 2,4, 7,14, and 21 d of treatment; in experiment 2, both aminoethoxyvinylglycine (AVG) and AgNO3 were applied to the horizontally-placed stems, and the cell numbers on sites of applications were measured after 40 d. Ethylene evolution from buds was found to be much greater in tilted seedlings than in upright ones. The cell numbers of wood fibers in shoots and 1-year-old stems were reduced in treatments with 12.5 x 10-7 μmol/L AVG, 12.5 x 10-8 μmol/L AVG, and 11.8 x 10-8 μmol/L AgNO3; whereas the horizontal and vertical diameters were reduced by treatment of 12.5 x 10-7 μnol/L AVG. Ethylene evolutions of shoots and 1-year-old stems were inhibited greatly in comparison with the control by applying 12.5 x 10-7 μmol/L AVG. The formation of a gelatinous layer of wood fibers was affected by neither AVG nor AgNO3 application. These results suggest that ethylene regulates the quantity of wood production, but does not affect G-layer formation in F. mandshurica Rupr. var. japonica Maxim, seedlings.     

干旱胁迫下接种丛枝菌根真菌对七子花非结构性碳水化合物积累及C、N、P化学计量特征的影响

以濒危植物七子花二年生幼苗为研究材料,采用盆栽试验方法,研究干旱胁迫和接种丛枝菌根真菌(AMF)处理对幼苗不同器官C、N、P化学计量关系和非结构性碳水化合物(NSC)含量的影响。试验共设计4个处理:对照(CK)、干旱胁迫(D)、接种AMF(AMF)、干旱胁迫和接种AMF(D+AMF)。结果表明: 在干旱胁迫下七子花根系AMF的侵染率显著下降,但接种AMF处理植株的株高、叶片数显著高于未接种处理。接种AMF显著提高了干旱胁迫下植株根、叶可溶性糖和NSC含量及茎、叶淀粉含量,且茎和叶可溶性糖与淀粉比显著下降。干旱胁迫导致植株C含量在根和叶中显著增加,P含量在茎中显著减少;与干旱胁迫相比,胁迫下接种AMF植株根、茎、叶P含量及叶C含量显著提高,而根C、N含量及茎C含量显著降低。胁迫下接种AMF植株根、茎C∶N、C∶P、N∶P和叶N∶P均显著低于单一胁迫处理。NSC与C∶N∶P计量比的相关性分析表明,根、叶P含量与可溶性糖和NSC含量呈显著正相关,茎P含量与淀粉和NSC含量呈显著正相关,各器官N∶P与NSC含量呈显著负相关。综上,干旱胁迫显著抑制了七子花幼苗的生长,接种AMF通过提高植株根和叶的可溶性糖含量、根的可溶性糖/淀粉,增加地上部分淀粉含量,促进P元素吸收和降低各器官N∶P来增强植株耐旱性,从而提高七子花幼苗在干旱环境中的存活率。     

Differentiation of human adipose-derived adult stem cells into neuronal tissue: Does it work?

Adipose tissue contains many cells and proteins that are of value not only for their potential therapeutic applications, but also for the low cost of their harvest and delivery. Mesenchymal stem cells (MSC) were originally isolated from the bone marrow, although similar populations have been isolated from adipose and other tissues. At one time, neural tissues were not regarded as regenerative populations of cells. Therefore, the identification of cell populations capable of neuronal differentiation has generated immense interest. Adipose tissue may represent an alternative source of cells that are capable of neuronal differentiation, potentially enhancing its use in the treatment of neurological disease. The aim of this review is to cover the current state of knowledge of the differentiation potential of human adipose-derived stem (ADAS) cells, specifically their ability to give rise to neuronal cells in vitro. This review presents and discusses different protocols used for inducing human ADAS cells to differentiate in vitro, and the neuronal markers utilized in each system.     

The control of in vitro direct main stem formation of Lilium longiflorum derived from receptacle culture, and rapid propagation by using in vitro stem nodes

The control of in vitro direct main stem formation by culturing receptacles, and a protocol for the micropropagation of Lilium longiflorum using in vitro main stem nodes derived from receptacle culture were developed. Receptacles from flowers cultured on MS medium containing 1.0 mg l^–1 gibberellic acid (GA₃) and 0.5 mg l^–1 6-benzyladenine (BA) resulted in direct main stem formation after 3 months culture. These stems were isolated and cut into nodal stem segments, which were then cultured on MS medium supplemented with 0.2 mg l^–1 BA. Shoots formed on each node after one month culture. These shoots were subcultured on MS medium containing 0.5 mg l^–1 BA for their mass propagation. An average of 30 vigorous and uniform shoots were formed per single shoot after each subculture. A cyclic and continuous system of propagation by multiplication of shoots was developed. Shoots were rooted on 1/2 MS medium containing 0.2 mg l^–1-naphthaleneacetic acid (NAA). One hundred plantlets that were acclimatized in the greenhouse had a 100% survival. A comparison was made with the traditional culture of explants derived from bulb-scales and with that from main stems.