影响喜树腋芽离体再生因素研究(英文)

本文以温室三年生喜树腋芽为外植体,研究了不同的基本培养基和基本培养基中添加不同浓度生长调节物质(BA 或TDZ)、蔗糖、琼脂以及培养基的pH 值对喜树腋芽分化的影响。结果表明:WPM 和B5 培养基适合喜树腋芽的诱导,MS 培养基不利于喜树腋芽的诱导。适合喜树腋芽增殖和分化的最佳生长调节物质为BA 1.0mg/L 或TDZ 0.1mg/L;最佳的蔗糖浓度为30g/L;最佳的琼脂浓度为6g/L;pH5.8-6.6 的范围均适合喜树腋芽的诱导,但最佳的pH 值为5.8。图1 表5 参14。     

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from <Emphasis Type="Italic">in vitro</Emphasis> cultured stem of <Emphasis Type="Italic">Populus alba</Emphasis>×<Emphasis Type="Italic">P. berolinensis</Emphasis>

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba~P berolinensis. Stems ofPopulus alba~P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.     

Multiplication of Elaeagnus angustifolia L. from a Single Shoot in Vitro

1INTRODUCTIONElaeagnusangusifoliaLisahighlytolerantnativewoodyspeciesinsouthernEuropeandAsia.Itmaybeusedinreforestationandsoil-waterconservationandforanexcellentwindbreaktoarrestwind(Bertrand,1985;LiShaozhong,etal,1997),materialsoffragranceandmedicineandnutritivefood(ChangZhaofeng,etal,1994;Goncharova,1997;Rasekh,1999;Ahmadiani2001;JiangFashou,etal,2002).Insomepreviousresearches,BA,2ip,KN(Bertrand,1985,Economou,1988)andBA(Iriondo1995;Mariella,1996)wereusedinshootmultiplicationofE…     

Direct shoot organogenesis from leaf explants of Embelia ribes Burm. f.: a vulnerable medicinal plant

An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Embelia ribes Burm. f., a vulnerable medicinal woody climber of the Western Ghats of India. The in vitro procedure involved three steps that included induction of shoot initials from leaf tissue, regeneration and elongation of shoots from the shoot initials, and rooting of shoots. The induction of shoot initials was achieved on Murashige and Skoog (MS) solid medium supplemented with different concentrations of thidiazuron (TDZ). The best medium for shoot induction was MS with 0.272 μM TDZ. Numerous shoot primordia developed within 2–3 weeks on the leaf margin as well as on the midrib region, without any callus phase. In the second step, the shoot clumps separated from the leaf explant on transfer to MS basal medium, resulting in the differentiation of 90% of the shoot initials into well-developed shoots. The 2- to 3-cm-long shoots rooted on half-strength MS basal medium supplemented with 4.90 μM indole-3-butyric acid (IBA) and 3% (w/v) sucrose in the third stage. The rooted plants could be established in soil with 70% success. This protocol could be utilized for in vitro propagation and conservation of this important threatened medicinal plant.     

楸树试管丛生芽继代增殖影响因素的研究

以楸树的幼嫩茎段为外植体诱导的腋芽丛生芽为试验材料,研究了不同因素对试管丛生芽继代增殖的影响.结果表明:继代增殖培养以MS+IBA 0.5 mg/L+BA 4.0 mg/L+PVP(100 mg/L)添加食糖30 g/L、琼脂0 g/L、pH为5.8的培养基为宜.食糖、葡萄糖和蔗糖对丛生芽增殖的影响没有明显区别.当培养基中不添加琼脂时,可以显著促进丛生芽增殖,而且有利于降低成长.     

银中杨茎段离体培养再生植株研究

以银中杨茎段为外植体,建立组织培养系统.试验结果表明,MS为最适基本培养基;各激素对诱导不定芽的作用大小为TDZ＞6-BA＞KT,诱导不定芽的最佳培养基为MS+0.1 mg·L-1TDZ+蔗糖20 g·L-1+琼脂0.7%;诱导不定芽再生的最佳光质为白光或黄光;生根培养基以1/2MS+0.5 mg·L-1IBA+蔗糖2...     

华富苹果离体叶片再生的主要影响因素研究

以花药培养选育的富士品种"华富"组培苗的叶片为试材,研究了离体叶片诱导再生不定芽过程中不同激素种类与组合、暗处理时间等因素对不定芽再生率的影响。结果表明:试管苗继代培养3～4代,取顶部嫩叶3～4片,剪除叶片边缘,叶片以近轴面接种到最佳诱导再生培养基MS+TDZ 2.0mg/L+IAA 0.75mg/L+蔗糖30g/L,pH值5.8,黑暗培养10～15d后,转入光照培养条件下,再生率达83.3%～86.7%。再生不定芽分化15～20d后转接到增殖培养基MS+BA 0.5mg/L+IBA 0.1mg/L上,当不定芽长度约2cm时再转接到生根培养基1/2MS+IAA 1.0mg/L+蔗糖2%,15d后,生根率可达95.0%以上。     

Factors influencing direct shoot regeneration from leaves,petioles, and plantlet roots of triploid hybrid <Emphasis Type="Italic">Populus</Emphasis> sect. <Emphasis Type="Italic">Tacamahaca</Emphasis>

Since the generation of full-sib artificial triploid families, rapid clone establishment and genetic improvements have been needed. Here, we report an in vitro method of direct shoot regeneration of a triploid hybrid poplar [(Populus simonii × P. nigra ‘Italica’) × (P. × ‘popularis’)]. Using different randomized block designs, we selected one triploid to evaluate the explant type, optimal concentrations of plant growth regulators and agar, and culture time under light or dark conditions over 60 days. The highest rate of shoot induction, 80.0%, was obtained using Murashige and Skoog (MS) medium supplemented with 0.2 mg/L benzyladenine, 0.04 mg/L naphthaleneacetic acid (NAA), and 5.5 g/L agar for the first 30 days in the dark, then 3 g/L agar for the next 30 days in light. This last medium yielded the best rate of shoot induction (6.32 shoots/explant). These three media were also used to evaluate the influence of the genotypes of the parents and hybrid triploids on regeneration. Two parents and three of the four full-sib triploids were regenerated successfully; different genotypes and explant types significantly affected the rate of shoot induction and average number of shoots. Leaves but not petioles were a suitable explant. One genotype produced the highest rate of shoot induction of 96.67%. Half-strength MS medium supplemented with 0.2 mg/L indole butyric acid and 0.04 mg/L NAA was the most effective for rooting; rooting rate was 96.67%, survival rate of transplants was 73.33%, and rooting frequency surpassed 85% for each genotype. Overall, this in vitro regeneration system will be useful for the propagation and genetic modification of triploid poplars.     

Micropropagation of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E. urophylla</Emphasis> AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.     

尾叶桉的组织培养及植株再生

研究了以尾叶桉(Eucalyptusurophylla)优树(U6无性系)无菌苗的叶子和茎段作为外植体诱导愈伤组织、丛生芽发生以及植株再生的过程。通过多种生长调节剂不同浓度组合的对比试验,确定了U6快繁体系的最适宜培养条件:(1)愈伤组织诱导培养基:MS 1-2mg/L2,4-D;(2)芽增殖培养基:MS 0.5mg/L6-BA;(3)生根培养基:1/2MS 2.0mg/LNAA。     

一品红组织培养技术研究

一品红组织培养以花轴序、叶片或顶芽、腋芽为外殖体，在MS BA2．0mg／L NAA0．50nm／L 蔗糖30g/L 琼脂5g/L CH70mg／L pH5．8的培养基上进行初代培养后转接到MS BA0．5 NAA0．20 蔗糖30g／L 琼脂5g／L。 pH5．8的培养基上扩繁，增殖率为13．1，丛生芽分化快；生长速度快，将2cm左右嫩梢剪下接种在1．／2MS NAA0．1 蔗糖15g/L 琼脂5mg／L pH5．8的培养基上诱导生根，10d左右即可生根。一品红组培苗生根与继代次数关系密切，继代次数10次以下，生根率达100％，之后逐渐下降，需要重新建立培养系。试验总结出一品红12个品种组织培养的一整套技术，为一品红工厂化生产提供了技术依据。     

西南桦节间茎段植株再生体系的建立

目的 探究西南桦间接器官发生过程的愈伤组织诱导、不定芽分化、生根诱导等各阶段的适宜培养基配方,建立西南桦高频再生体系,为其遗传转化和良种繁育提供理论依据和技术支持。 方法 以西南桦TC2号无性系的节间茎段为外植体开展愈伤组织诱导、不定芽分化以及生根培养基筛选试验,并优化了预培养条件,揭示了愈伤组织诱导阶段基本培养基、激素浓度、暗培养时间,愈伤组织分化阶段激素组合,以及预培养条件等因素对不定芽分化的影响。 结果 (1) 预培养的适宜条件为：弱光培养 (1 000 lx) 15 d后转至暗培养7 d,再正常光照 (2 000 lx) 培养8 d,该条件下可获得适度黄化的植株,其平均株高和节间长度分别为6.6 cm和3.1 cm; (2) 适合西南桦节间茎段愈伤组织诱导的培养基为WPB5 + 1.0 mg·L−1 TDZ + 0.2 mg·L−1 NAA + 20 g·L−1蔗糖 + 5.8 g·L−1琼脂 (pH5.8),适宜暗培养时间为15 d;(3) 适合愈伤组织分化的培养基为WPM + 0.8 mg·L−1 6-BA + 0.5 mg·L−1 GA3 + 30 g·L−1蔗糖 + 5.8 g·L−1琼脂 (pH5.8);(4) 采用上述最优方案,其茎段诱导的愈伤组织分化率和净增殖系数为88.9%和6.2以上,平均每个预培养植株可产生56.8个不定芽;(5) 适宜生根培养基为WPM + 0.1 mg·L−1 NAA + 20 g·L−1蔗糖 + 5.8 g·L−1琼脂 (pH5.8),培养30 d后生根率可达100%。 结论 本研究完整构建了高效稳定、重复性好的西南桦节间茎段高频再生体系,该体系不仅愈伤组织分化率和增殖系数较高,还可提升节间茎段取材的效率,为今后开展西南桦组培快繁以及利用基因工程进行其遗传性状改良奠定了基础。     

Direct plant regeneration from nodal explants of <Emphasis Type="Italic">Balanites aegyptiaca</Emphasis> L. (Del.): a valuable medicinal tree

This report describes in vitro shoot induction and plant regeneration from nodal segments of Balanites aegyptiaca on Murashige and Skoog (MS) medium fortified with 6-benzyladenine (BA), thidiazuron (TDZ) and kinetin (Kin) (0.5–20.0 μM). MS medium supplemented with BA (12.5 μM) was the most effective in inducing bud break and growth and also in initiating multiple shoot proliferation. However, the optimal level of TDZ supplementation to the culture medium was 5.0 μM. Shoot cultures were established by repeatedly subculturing the original nodal explants on the same medium. Highest number of shoots (11.5 ± 0.7) and shoot length (5.0 ± 0.2 cm) were achieved when cultures were subcultured on MS medium supplemented with 12.5 μM BA and 1.0 μM α-naphthalene acetic acid (NAA). The shoots regenerated from TDZ supplemented medium when subcultured to hormone free MS basal medium considerably increased the rate of shoot multiplication and shoot length by the end of fifth subculture. Rooting of the shoots was achieved on MS medium augmented with 1.0 μM indole-3-butyric acid (IBA) plus 0.5% activated charcoal followed by their transfer to half strength MS basal medium. The in vitro raised plantlets with well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse with 70% survival rate. The results of this study provide the first successful report on in vitro direct plant regeneration of B. aegyptiaca.     

Shoot multiplication and plant regeneration in <Emphasis Type="Italic">Caragana fruticosa</Emphasis> (Pall.) Besser

Different nutrient media can affect in vitro culturing protocols, and experimentation under varied growth conditions is valuable in plants where in vitro methods are in preliminary stages. We carried out the first in vitro propagation studies for the endangered species Caragana fruticosa (Fabaceae). We evaluated various nutrient media for their impact on shoot elongation and axillary bud proliferation using different concentrations of 6-benzylaminopurine (BA) and α-naphthaleneacetic acid (NAA). Shoot elongation was evaluated based on adventitious shoot primary culture and subculture regeneration from Caragana seedlings. Our goal was to improve both micropropagation and regeneration in C. fruticosa. MS nutrient media was superior to 1/2MS macronutrients, DKW, QL, and WPM for shoot elongation and axillary shoot proliferation. Shoots grown on 1/2MS and WPM exhibited some chlorosis, and shoots on QL produced larger leavers than plants growing on normal medium. The shoot proliferation coefficient on MS media supplemented with 2.22 μM BA and 0.44 μM BA + 2.69 μM NAA was significantly higher than that with other treatments in the primary culture. Shoots on 2.22 μM BA showed a higher proliferation coefficient (3.17) than others in the subculture. Shoots were rooted on 1/2MS medium with the addition of different concentrations of NAA. The optimal concentration for rooting was 0.27 μM NAA (74%). Roots exhibited many stout and long root hairs. Survivl of established plantlets was 82% at 30 days after transfer to soil. Plants established in the green house showed normal growth and displayed no apparent morphological differences compared to stock plants.     

In vitro plant regeneration system for <Emphasis Type="Italic">Cassia siamea</Emphasis> Lam., a leguminous tree of economic importance

A method for rapid in vitro propagation of Cassia siamea Lam. using cotyledonary node explants, excised from 14-day old aseptic seedlings, has been established. Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) singly or in combination with auxins was used for regeneration studies. Among the single treatment of three cytokinins BA at 1.0 μM was found to be optimum for direct shoot regeneration as it induced an average of 8.20 ± 0.66 shoots per explant. The regeneration frequency further enhanced with the application of auxin along with optimal BA concentration. The highest frequency for shoot regeneration (90%), the maximum number of shoots per explant (12.20 ± 0.73) and the maximum shoot length (6.40 ± 0.07) cm were obtained on the medium consisted of MS + 1.0 μM BA + 0.5 μM NAA. Successful in vitro rooting was induced from cut end of the microshoots when placed on half-strength MS + IBA (2.5 μM). The regenerated shoots with well developed root system were successfully acclimatized and established in pots containing sterilized garden soil and garden manure (1:1) and grown under greenhouse conditions with 85% survival rate.     

白刺花胚性愈伤组织诱导及体细胞胚发生

【目的】探讨不同植物生长调节剂对白刺花胚性愈伤组织诱导的作用,以及培养基中氮源和无机盐浓度对白刺花体细胞胚发生和植株再生的影响,以期建立白刺花体细胞胚发生、发育及调控技术体系,为白刺花种苗快速繁殖体系建立及遗传转化研究提供参考。【方法】以白刺花叶片为外植体,研究生长调节剂2,4-D(1.0、2.0、3.0、4.0 mg ·L -1 )、NAA(0、0.5、0.8、1.0 mg ·L -1 )、6-BA(0.2、0.5、1.0、2.0 mg ·L -1 )和TDZ(0、0.2、0.5、1.0 mg ·L -1 )组合对胚性愈伤组织诱导,及NAA(0、0.2、0.5 mg ·L -1 )、6-BA(0、0.5、1.0 mg ·L -1 )和TDZ(0、0.2、0.5 mg ·L -1 )组合对体细胞胚发生的调控作用,筛选最优生长调节剂组合;并研究培养基中KNO 3和NH 4NO 3比例对体细胞胚发生的作用,及MS培养基中无机盐浓度(1/5MS 、1/4MS、1/3MS、1/2MS)对体细胞胚萌发的影响,筛选最佳的体细胞胚发育及成熟萌发条件。【结果】白刺花叶片外植体胚性愈伤组织诱导适宜培养基为MS + 2,4-D 3.0 mg ·L -1 + NAA 0.5 mg ·L -1 + 6-BA 0.2 mg ·L -1 + TDZ 1.0 mg ·L -1 +蔗糖40 g ·L -1 +琼脂7.0 g ·L -1 ,诱导率为42.0%。采用MS基本培养基时,最佳的体细胞胚发生培养基为MS + NAA 0.5 mg ·L -1 + 6-BA 1.0 mg ·L -1 + TDZ 0.5 mg ·L -1 +蔗糖40 g ·L -1 +谷氨酰胺100 mg ·L -1 +琼脂7.0 g ·L -1 ,体细胞胚发生率为78.46%,总胚数为对照的3.6倍;MS培养基中,KNO 3浓度提高1倍、NH 4NO 3降至1/2时,体细胞胚发生率可提高至91.33%,总胚数为采用MS基本培养基时的1.4倍;1/3MS培养基有利于体细胞胚的萌发,萌发率为82.75%,幼苗长势良好,单株平均鲜质量为76 mg,幼苗驯化移栽1个月后成活率达90%以上。【结论】白刺花叶片接种于添加2,4-D、NAA、6-BA和TDZ不同组合的诱导培养基上,可脱分化形成愈伤组织或胚性愈伤组织,2,4-D浓度对愈伤组织形态和质地有较大影响。TDZ有利于体细胞胚的形成,适宜浓度的生长素与细胞分裂素组合及硝态氮和铵态氮的比例对体细胞胚的形成和发育具有调控作用,降低MS无机盐浓度可提高体细胞胚萌发率,本试验体系的再生植株移栽成活率达90%以上。     

粗皮桉再生系统的研究(英文)

该研究在国内外首次报导了粗皮桉的再生系统 .利用粗皮桉的外植体通过两种不同的再生途径建立了高效的再生系统 .首先探讨了粗皮桉的下胚轴和子叶的体细胞胚胎发生 .研究发现 ,下胚轴和子叶在含有 2 ,4 D(添加有BA或激动素 )的培养基上可诱导三种典型不同的愈伤类型 .其中 ,来源于下胚轴的颗粒状的、松脆的胚性愈伤在分化培养基上可获得高频率的再生小植株 .讨论了不同类型的愈伤组织和其对应的分化再生率之间的关系 .此外 ,本研究探讨了来源于粗皮桉的下胚轴、子叶及成熟叶片在添加有一系列BA和NAA的不同浓度组合的B5培养基上的器官发生途径 ,三种不同的外植体在不同的再生培养基上的最高再生频率不同 ,其中成熟叶片、子叶和下胚轴的最高再生率逐渐提高 ,分别为 4 1 6 7%、6 6 6 7%和 83 33% .该文还讨论了两种不同再生系统的意义     

An efficient in vitro process for recurrent production of cloned plants of Vitex negundo L

An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and ??-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 ??M, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4?C5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 ??M indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants.     

Modulation of in vitro morphogenesis in nodal segments of Salix tetrasperma Roxb. through the use of TDZ, different media types and culture regimes

A comparative performance of two different media formulations (woody plant medium (WPM) and Murashige and Skoog??s (MS) medium) for their ability to inflict in vitro shoot development in nodal segments of Salix tetrasperma Roxb. has been carried out. Thidiazuron (TDZ) in various concentrations was used as a supplement to the basal media. Media types, TDZ concentrations, exposure duration and culture regimes played an important role in affecting multiple shoot production. WPM supplemented with 2.5???M TDZ for 4?weeks exposure was found to be the best for maximum (4.53?±?0.27) shoots production in vitro. Transfer to a secondary medium consisting of 6-benzyladenine (1.0???M) and ??-naphthalene acetic acid (0.5???M) enhanced the multiplication rate and by the end of 12?weeks, 20.33?±?0.33 shoots with shoot length, 4.70?±?0.26?cm were produced on WPM. Rooting of the regenerated shoots was achieved on half strength basal media (either WPM or MS) containing 0.5???M indole-3-butyric acid. In all the experiments, different growth parameters were scored and WPM was found to be superior to MS medium. The regenerated plantlets were successfully acclimatized in the field with about 81?% survival.     

Rapid clonal multiplication of a woody tree, <Emphasis Type="Italic">Vitex negundo</Emphasis> L. through axillary shoots proliferation

An efficient and improved shoot regeneration technique for the micropropagation of Vitex negundo, an aromatic and medicinal shrub through in vitro culture of nodal segments with axillary buds, is described. Thidiazuron (TDZ) used at 1.0 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 25 microshoots per nodal explant with axillary buds, after 4 weeks of culture. By repeated subculturing of nodal explants, a high-frequency multiplication rate was established. Optimum shoot multiplication and elongation was achieved when TDZ exposed explants were subcultured on Murashige and Skoog (MS) media containing a combination of 1.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA). Efficient rooting was achieved directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 10 min which was the most effective in inducing roots, as 97% of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered. No phenotypical differences for morphogenesis were observed among the regenerants.