In vitro culture of Aloe Barbadensis Mill.: Micropropagation from vegetative meristems

A method for rapid and highly effective plant micropropagation from vegetative meristems was established for Aloe barbadensis Mill. Plant micropropagation was achieved culturing apices on medium containing 1.1 M 2,4-dichlorophenoxyacetic acid and 2.3 M kinetin for 15–30 days. High morphogenetic ability was maintained by transferring explants (after 60 days) on media containing 0.11 M 2,4-dichlorophenoxyacetic acid and 2.2 M 6-benzylaminopurine.     

Clonal propagation of papaya in vitro

A procedure for the rapid tissue culture propagation of papaya is being developed. Tissue culture methods using apices of nursery and orchard trees of Carica papaya cv. Sunrise Solo were evaluated. The explants were established in a modified Murashige and Tucker (1969) basal medium with half-strength inorganic salts, 0.5mgl^-1 6-benzylaminopurine (BA) and 0.2mgl^-1 naphthaleneacetic acid (NAA). Established explants were transferred to a proliferation medium consisting of Murashige and Tucker (1969) basal medium, 0.5mgl^-1 BA and 0.1mgl^-1 NAA, which caused extensive multiplication of shoots. Rooting was induced at a higher frequency by subculturing plantlets onto media with indole-3-butyric acid (IBA) than with NAA.     

Rapid propagation of agave by in vitro tissue culture

A procedure for rapid propagation of Agave (A. cantala Roxb., A. fourcroydes Lem. and A. sisalana Perrine, (Agavaceae) have been developed. The explants were excised from stolon plantlets, sterilized and cultivated on Murashige and Skoog (MS) basal medium containing 2% sucrose, 10% coconut water and 0.8% agar. The addition of following combination of growth substances—0.075 mgl^-1 naphthalenacetic acid (NAA)+0.1 mgl^-1 indolylbutyric acid (IBA)+0.5 mgl^-1 kinetin (KIN) caused an extensive proliferation of multiple shoot primordia. Subcultures of these on the same medium were successful for the multiplication with an index of 3–4 times per 4 weeks subculture period. Shoots were rooted on hormone free MS medium and then transferred into a sand bed for acclimation before field planting.     

Rapid clonal propagation of Dioscorea zingiberensis

A protocol was developed for rapid in vitro propagation of Dioscorea zingiberensis Wright using stems as explants. MS medium with the macroelements at half strength and supplemented with 20.0 g l^–1 sucrose and 8.0 g l^–1 agar was used as basal medium. Lateral buds on nodal cuttings grew into shoots within 20 days after culture on basal medium supplemented with 4.4 M 6-benzylaminopurine (BAP) and 1.1 M -naphthalene acetic acid (NAA). The shoots were cut into segments and cultured on medium with 8.9 M BA and 5.4 M NAA for 30 days for callus formation. The callus was cut into pieces and cultured on medium containing 22.2 M BAP and 1.1 M NAA, on which 87.5% of the callus pieces regenerated multiple shoots within 50 days. The shoots were rooted on medium containing 4.9 M indole-3-butyric acid (IBA) for 20 days. Adventitious buds and shoots could be repeatedly formed after the calli were cut into pieces and cultured on the medium containing 8.9 M BAP plus 1.1 M NAA. More than 85% of the regenerated plantlets survived and grew vigorously 1 month after they were transplanted in vermiculite and each plant formed 2–4 microtubers 3 months of transplanting.     

In vitro propagation of Vicia faba L. by micro-cutting and multiple shoot induction

The influences of nitrogen sources, culture temperature and activated charcoal supplements were studied in relation to the rooting ability of V. faba cuttings. The interaction of these factors led to quantitative and qualitative modifications of the culture responses. Low temperatures (14–18°C) were suitable for in vitro culture, limiting the formation of phenolics in plant material and making activated charcoal supplement unnecessary. Nitrogen supplements contributed in modifying the different plant responses, in accordance with temperature. Multiple shoot formation was obtained from the cotyledonary node and from the stem nodes cultivated in the presence of 6-benzylaminopurine (BAP). BAP at 4 mg l^-1 was the most effective concentration in promoting high rates of shoot development. The original position of stem nodes was found to determine the explant response to plant growth regulator treatments, possibly due to the effect of residual apical dominance.     

Theobroma cacao L.: an axillary bud in vitro propagation procedure

An axillary bud derived in vitro propagation procedure for Theobroma cacao is described. The method allows bud expansion and elongation, including leaf development, as early as 13 days after placing explants on establishment medium. Shoots have been maintained in culture vessels for six months with little evidence of decline. Propagules have been induced to form roots, hardened off, and moved to the greenhouse within six weeks of initial explant establishment. Some plants have been in the greenhouse for twelve months and are growing in an apparently normal manner. No major differences in response from UF-667 or EQX-100 derived seed-grown trees were found suggesting that the method may not be limited by genotype. Long-term growth without added plant growth regulators has been seen in over 2400 explants.Abbreviations NAA naphthaleneacetic acid - IBA indolebutyric acid - BAP benzylaminopurine - WPM Lloyd-McCown woody plant medium Published as paper No. 8148, Journal Series, Pennsylvania Agricultural Experiment Station     

木立芦荟组织培养pH分化特性及快速繁殖的研究

以木立芦荟的叶片、叶鞘、带腋芽的茎段为外植体进行试管培养 ,结果叶鞘和茎段可诱导形成愈伤组织 ,腋芽直接萌生。经试验筛选出各培养阶段最适宜的培养基为 :( 1 )愈伤组织诱导 ,MS BA 2 .5mg/L NAA0 .1 5mg/L ;( 2 )腋芽萌生 ,MS BA 2 .0mg/L NAA 0 .1 5mg/L ;( 3 )丛生芽分化及继代 ,MS BA 2 .0mg/L NAA0 .1 0mg/L ;( 4)生根 ,MS BA 0 .3～ 0 .5mg/L IBA 0 .2mg/L 活性碳 0 .5%。研究还发现 ,培养基酸碱度对木立芦荟组织培养分化效果的影响非常显著。     

Rapid in vitro propagation,conservation and analysis of genetic stability of Viola pilosa

A protocol for in vitro propagation was developed for Viola pilosa, a plant of immense medicinal value. To start with in vitro propagation, the sterilized explants (buds) were cultured on MS basal medium supplemented with various concentrations of growth regulators. One of the medium compositions MS basal + 0.5 mg/l BA + 0.5 mg/l TDZ + 0.5 mg/l GA₃ gave best results for in vitro shoot bud establishment. Although the problem of shoot vitrification occurred on this medium but this was overcome by transferring the vitrified shoots on MS medium supplemented with 1 mg/l BA and 0.25 mg/l Kn. The same medium was found to be the best medium for further in vitro shoot multiplication. 100 % root induction from in vitro grown shoots was obtained on half strength MS medium supplemented with 1 mg/l IBA. In vitro formed plantlets were hardened and transferred to soil with 83 % survival. Additionally, conservation of in vitro multiplying shoots was also attempted using two different approaches namely slowing down the growth at low temperature and cryopreservation following vitrification. At low temperature retrieval rate was better at 10 °C than at 4 °C after conservation of in vitro multiplying shoots. In cryopreservation–vitrification studies, the vitrified shoot buds gave maximum retrieval of 41.66 % when they were precooled at 4 °C, while only 16.66 % vitrified shoots were retrieved from those precooled at 10 °C. Genetic stability of the in vitro grown plants was analysed by RAPD and ISSR markers which indicated no somaclonal variation among in vitro grown plants demonstrating the feasibility of using the protocol without any adverse genetical effects.     

Novel bioactive maloyl glucans from aloe vera gel: isolation, structure elucidation and in vitro bioassays

In this study, three novel maloyl glucans were isolated at temperatures below 15 degrees C from aloe vera gel (Aloe barbadensis Miller). These compounds were characterized using NMR spectroscopy, ESIMS, MALDITOF-MS and capillary electrophoresis. The compounds were characterized as 6-O-(1-L-maloyl)-alpha-,beta-D-Glcp (veracylglucan A), alpha-D-Glcp-(1-->4)-6-O-(1-L-maloyl)-alpha,-beta,-D-Glcp (veracylglucan B) and alpha-D-Glcp-(1-->4)-tetra-[6-O-(1-L-maloyl)-alpha-D-Glcp-(1-->4)]-6-O-(1-L-maloyl)-alpha,-beta-D-Glcp (veracylglucan C). These unusual malic acid acylated carbohydrates were then tested in vitro for effects on cell proliferation and gene expression of proinflammatory cytokines, IL-6, IL-8 and ICAM-1, using RT-PCR. Veracylglucan B demonstrated potent anti-inflammatory and anti-proliferative effects, while Veracylglucan C, on the other hand, exhibited significant cell proliferative and anti-inflammatory activities. Veracylglucan A could only be isolated in smaller quantities, and it proved to be very unstable. Thus no biological effects could be observed in this respect. The in vitro bioassays also indicated that Veracylglucan B and C are antagonistic and competitive in their effects on cell proliferation. The results of this work represent a major step forward in the research on aloe vera gel. This is the first time that two fully chemically characterized compounds are shown to be responsible for known biological activities of aloe vera gel.     

Vegetative propagation of Begonia venosa Skan in vitro from inflorescence explants

Methods are described for the vegetative propagation of Begonia venosa Skan. Young flower buds are capable of producing callus which, contrasting to callus from leaves of adult plants, is very organogenic. For callus induction are required: BA and NAA at a conc. of 0.5 mgl^–1, 21 °C and low irradiance. Subculture of organogenic callus is optimal on a medium with 0.1 mgl^–1 BA and 2% glucose, whereas NAA is ommitted. Shoot development from preformed adventitious buds is enhanced by lowering the BA and glucose conc. Optimal rooting of excised shoots is obtained on a medium without regulators and a low glucose level. No visible mutations can be detected in the plant material, even not at the flowering stage.Abbreviations (BA) 6-Benzylaminopurine - (2iP) N⁶-(2-isopentenyl)adenine - (MS)(1) Murashige and Skoog - (NAA) 1-Naphthaleneacetic acid Publ. 535     

In vitro propagation of the alkaloid producing plant Datura insignis Barb. Rodr.

A method is presented for the in vitro propagation of Datura insignis Barb. Rodr. Nodal explants were cultured on Murashige and Skoog medium supplemented either with BA alone or in combination with 2,4-D or IAA. Shoot multiplication and elongation were obtained in various growth regulator concentrations. Best results were obtained in a medium with 1.0 mgl^-1 of BA. Individual shoots were excised and transfered to growth regulator-free medium for rooting. Additional multiplication was obtained by single-node culture using explants from these in vitro rooted shoots. Rooted plantlets were successfully grown in soil.     

In vitro tuberization in white yam (Dioscorea rotundata Poir)

Nodal cuttings of white yam were induced to produce microtubers on a MS-revised medium supplemented with various concentrations of sucrose, 20 mgl^–1 L-cysteine, 0.5 mgl^–1 kinetin and 0.7% agar. The frequency of tuberization was affected by the daylength, which is optimal at 12 and 16 h of light depending on the sucrose concentration. The microtubers were planted in a seed bed and grown to maturity. The importance of in vitro tuberization of yam as a means of international germplasm distribution or exchange as well as for the propagation of planting material is discussed.     

海南龙血树离体快速繁殖

以3月龄高约25cm的海南龙血树为基本材料,以茎节纵切块为外植体,在改良MS 6-BA2.0mg/L NAA0.1mg/L培养基中培养1个月后萌芽率为78.6%。萌芽在MS 6-BA1.5mg/L NAA0.2mg/L培养基中连续两次继代培养,2个月后芽增殖倍数为2.9。丛芽及单芽都在MS 6-BA1.0mg/L NAA0.2mg/L培养基中进行培养,1个月后丛芽的单芽增殖倍数为4.6,单芽增殖倍数为0.8。萌芽放入MS(改良) NAA0.2mg/L培养基中进行培养,生根率为100%,移栽成活率为100%。     

Requirements for in vitro propagation of seven nitrogen-fixing Alnus species

Studies on the in vitro propagation of Alnus crispa, A. glutinosa, A. incana, A. japonica, A. rubra, A. sinuata and A. viridis indicated interspecific as well as intraspecific variations in their requirements for in vitro culture. The WPM and Blaydes media supported, respectively, growth of A. glutinosa and A. crispa but not that of both species, while the MS medium induced equal or significantly better growth than WPM and Blaydes media for both species. The optimum type and concentration of sugar to be used in the multiplication medium varied with species. Only A. glutinosa showed good growth on sucrose while glucose was optimum for all other species but at different concentrations. All species rooted in 3 weeks on half-strength MS medium including 1 M IBA. All clones of A. glutinosa and A. rubra rooted 100%, whereas easy-to-root and difficult-to-root clones were observed in the other species. In the rooting medium, glucose promoted rooting of the difficult-to-root clones better than sucrose. Survival following transfer to an artificial substrate was 100% for all species. Nodulation tests using pure cultures of two Frankia strains showed 100% nodulation on all Alnus clones.     

In vitro clonal propagation of Pisum sativum L.

A system of in vitro clonal propagation has been developed in Pisum sativum L. (cv. Bohatýr). A modified MS-medium supplemented with 20 M 6-benzylaminopurine (BAP) and 0.1 M -naphthaleneacetic acid (NAA) was used to induce multiple shoot formation from shoot apices, axillary buds of the first normal leaf, axillary buds of the first and second primary scales and axillary buds of cotyledons of 4 to 6 day old pea seedlings. Meristem explants maintained a high proliferation ability in each subculture in the course of 20 months of the culture. Regenerated shoots were rooted in the same basal medium containing 5 M NAA. Rooted plants were cultured in hydroponic pots filled with half-strength MS-medium to attain anthesis and seed maturity. The phenotypic uniformity of the regenerants was evaluated. Cytological investigation confirmed the diploid stage (2n=14) of regenerants and their progeny. Histological studies revealed that proliferating shoots originated from axillary and adventitious buds. In vitro propagation is discussed as related to pea breeding.     

Factors affecting the establishment and maintenance of ‘Titan’ red raspberry root organ cultures

Root organ cultures of red raspberry (Rubus ideaus cv. Titan) were established from two cm long terminal segments of adventitious roots induced in vitro on micropropagated shoot cultures. Growth of root organ cultures was almost entirely by initiation and elongation of lateral roots rather than by terminal elongation from the apical meristem of the original root explants. Indolebutyric acid (IBA) was required for lateral root initiation and elongation. The optimal IBA concentration for lateral root initiation decreased from 0.5 to 0.1 mg/L from the first to the fifth passage in culture. Two cm root explants initiated more lateral roots per unit length than four cm explants. Liquid Anderson's medium was superior to other basal nutrient and vitamin formulations tested. Root growth in liquid culture was stimulated by aeration. Root organ cultures also grew on media solidified with gelrite or agar. A rapid transfer technique was developed for subculturing these which involved cutting and transferring 1 cm discs of roots and underlying medium. Adventitious bud formation occurred spontaneously, but sporadically in liquid cultures, and was not influenced by cytokinin, auxin (spat) light, or chilling.     

In vitro propagation of green-foliaged Dracaena fragrans Ker.

Callus tissue was induced in young stem segments cultured on MS based media supplemented with 0.25–0.5 mg l^-1 2, 4-D. Shoots were differentiated on media containing 0.5–1.0 mg l^-1 BA and 0.5–2.0 mg l^-1 IBA or 0.1–0.2 mg l^-1 NAA. The same media were suitable for shoot multiplication. Shoot elongation and rooting were strongly inhibited by BA and stimulated by auxins IBA and NAA. Medium containing 0.5 mg l^-1 IBA was optimal for rooting. Root elongation was stimulated by light and inhibited in darkness. Transfer of rooted plantlets to outdoor conditions was feasible and special hardening procedures were not required. Among more than 5000 plants produced by this procedure only 9 off-type plants with variegated leaves were found.     

褐纹报春苣苔组织培养与快速繁殖

褐纹报春苣苔(Primulina glandaceistriata)是一种极具观赏价值的喀斯特地区野生花卉,目前尚未有褐纹报春苣苔组培快繁的研究报道。该研究以褐纹报春苣苔的叶片为外植体,通过两种途径建立其组培快繁体系。结果表明:适宜的不定芽诱导培养基为MS+6-BA 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1),适宜的不定芽增殖培养基为MS+ZT 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1)+活性炭0.05 g·L~(-1),增殖系数为11.09;适宜的愈伤组织诱导培养基为MS+TDZ 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1),适宜的愈伤组织分化培养基为MS+ZT 1.0 mg·L~(-1)+NAA 0.10mg·L~(-1),分化系数为12.46;适宜的生根培养基为1/2MS+IBA 0.1 mg·L~(-1)+活性炭0.05 g·L~(-1)或1/2MS+IBA0.5 mg·L~(-1)+活性炭0.05 g·L~(-1),生根率为100%。该研究结果成功建立了褐纹报春苣苔的组培快繁体系,为今后褐纹报春苣苔的种苗繁殖和遗传转化提供了技术支持。     

青钱柳的快速繁殖技术

该文以青钱柳幼嫩茎段为外植体,研究其种苗快速繁殖技术。结果表明:青钱柳最佳采样时间为4月—6月,最佳外植体为轻微木质化茎段;最佳的外植体表面消毒方法为用0.1%HgCl₂浸泡5～7 min,消毒成功率54.1%,无菌外植体存活率88.7%;初代芽诱导培养基为MS+6-BA 2.0 mg·L^-1+IBA 0.2 mg·L^-1+蔗糖30.0 g·L^-1,芽诱导率80.5%,培养21 d初代芽苗平均高度3.0 cm;最佳继代增殖培养基为MS+6-BA 0.5mg·L^-1+IBA 0.05 mg·L^-1+TIBA 0.02 mg·L^-1+蔗糖30.0 g·L^-1,培养35 d,增殖系数7.0/35 d,平均苗高4.5cm,芽苗健壮且无玻璃化;生根前的壮苗培养基为MS+6-BA 0.5 mg·L^-1+IBA 0.05 mg·L^-1+蔗糖30.0 g·L^-1,...     

木薯离体培养与快速繁殖

选取3个木薯品种的腋芽作为外植体,分别接种于MS基本培养基上进行离体培养及快速繁殖。结果表明, 6-BA不同浓度对木薯品种不定芽诱导效果不一,在MS+6-BA 0.5~1.0mg/L中,3个品种均可诱导产生幼芽;在MS+6-BA 3.0mg/L中,3个品种的幼芽诱导率高达100%;MS+6-BA 3.0mg/L+NAA 0.1mg/L对继代效果较好;MS+NAA 0.1mg/L对生根效果最佳。