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.     

In vitro clonal propagation of dioecious Carica papaya

A procedure for in vitro propagation of dioecious papaya clones is described. A high rate of success in culture estbalishment was obtained when axillary buds were taken from lateral shoots of hedged rooted cuttings grown in a greenhouse. Seasonal endophytic contamination was suppressed by shaking propagules for 24 h in 300 mgl^-1 rifampicin or by incorporating it at 50 mgl^-1 into the medium. Murashige & Skoog (MS) basal medium supplemented with 0.5 mgl^-1 6-benzyladenine and 0.1 mgl^-1 naphthaleneacetic acid was used for establishment and proliferation. The addition of 160 mgl^-1 adenine sulfate improved multiplication and shoot growth. An elongation stage on MS medium supplemented with 1.0 mgl^-1 kinetin and 0.05 mgl^-1 naphthaleneacetic acid was necessary before rooting. Rooting was obtained at a high rate on half-strength macroelements of MS medium supplemented with 1.0 mgl^-1 indole-3-butyric acid. Commercial plots of papaya plants obtained through this procedure already exist.     

In vitro propagation of Ancistrocladus abbreviatus Airy Shaw (Ancistrocladaceae)

Ancistrocladus abbreviatus Airy Shaw (Ancistrocladaceae), a West African liana producing naphthylisoquinoline alkaloids, was successfully raised from seeds in vitro. Clonal propagation was best achieved growing nodal stem segments on 1/5 Linsmaier and Skoog medium with full strength organics and supplemented with 0.02 μM thidiazuron, 4.44 μM 6-benzylaminopurine and 0.05 μM 1-naphthaleneacetic acid. Detached axillary shoots were grown on Anderson's Rhododendron medium devoid of phytohormones and rooted within one month when dipped in 4.92 μM indole-3-butyric acid. Rooted plants became acclimatized to nonsterile greenhouse conditions. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro shoot tip multiplication of cowpea Vigna unguiculata (L.) walp

Summary Shoot multiplication was induced in cowpea, cv. Georgia-21, from shoot tip explants. Shoot tips, 5 mm long, were isolated from in vitro-grown seedlings and cultured on MS medium containing N⁶-benzyladenine (BA) at 1, 2.5, or 5 mg/liter (4.4, 11.1, or 22.2 μM) or 6-furfurylaminopurine (kinetin) at 1, 2.5, or 5 mg/liter (4.6, 11.6, or 23.2 μM) combined with 2,4-dichlorophenoxyacetic acid (2,4-D) at 0.01, 0.1, or 0.5 mg/liter (0.05, 0.5, or 2.3 μM) or naphthaleneacetic acid (NAA) at 0.01, 0.1, or 0.5 mg/liter (0.05, 0.5, or 2.7 μM). Cultures were maintained at a 12-h photoperiod (40 μmol·m⁻²·s⁻¹) and 23 ± 2° C. Treatments with BA induced greater shoot proliferation than those with kinetin. The highest number of shoots was produced on 5 mg (22.2 μM) BA per liter in combination with NAA or 2,4-D at 0.01 mg/liter (0.05 μM). Callus proliferated from the basal ends of shoot pieces in all treatments. The cultures also formed roots in the presence of kinetin, but not on BA-containing medium. To produce whole plants, the shoots were separated and rooted on 0.1 mg (0.5 μM) NAA per liter. Resulting plants grew normally under greenhouse conditions. Shoot tips provide an excellent explant source for cowpea micropropagation and can be used for callus induction.     

小叶兜兰的种子发育和无菌萌发

为建立小叶兜兰的繁育技术体系,本研究通过无菌播种的方法,辅以TTc生活力测定等方法,比较了小叶兜兰种子在授粉后不同发育时期和培养条件下的萌发率,对小叶兜兰种胚的发育过程进行了显微观察,探讨种胚发育程度与萌发的关系。结果表明,小叶兜兰种胚的发育阶段对萌发的影响最大,授粉后255d的种子萌发率最高（90．71％）,该阶段种子仍呈白色但微干燥,种胚刚发育至球形胚阶段,胚柄尚存。1／4MS和1／2MS为小叶兜兰适宜的基本培养基,添加100mg·L^-1的土豆汁对小叶兜兰的无菌萌发有良好的促进作用。     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。     

Asymbiotic Seed Germination and in vitro Seedling Rapid Development of Paphiopedilum spicerianum

In this paper, we studied the asymbiotic seed germination and seedling development of Paphiopedilum spicerianum, a wild plant with extremely small populations (PSESP) in China. By analogizing the influences of different media, seed maturity, pretreatments and lights on the seed germination, the results showed that the best germination condition is using the seeds at 270 days after pollination, pretreating with 1% NaOCl for 40 min, on the 1/4MS+10% coconut water medium under 24 hours darkness for 4 weeks. For seedling formation, 30g·L-1 Hyponex No 1 medium with 10mg·L-1 α naphthlene acetic acid, 05g·L-1 activated charcoal and 10% banana homogenate was the most effective. For seedling development, the same medium used for seedling formation with supplemented 10g·L-1 6 benzyladenine was most suitable.     

In vitro propagation of Amaryllis belladonna

Amaryllis belladonna L. plants were multiplied successfully by means of tissue culture techniques. Different plant parts were tested as explant material, but plantlets could only be generated from the twin-scales and immature scapes. These in vitro-formed plantlets were divided into four parts and used for further multiplication. The twin-scale explants had the highest multiplication rate when a medium with 22.2 M benzyladenine and 0.54 M naphthaleneacetic acid was used. The sucrose concentration played an important role in the initiation of new plantlets, and the best results were obtained when a sucrose concentration of 2–3% was used. Anatomical observations were made during the initiation of the new plantlets.Abbreviations BA benzyladenine - NAA naphthaleneacetic acid - Benomyl (methyl [1-[(butylamino) carbonyl]-1H-benzimidazol-2-yl] carbamate) - Folpet (2-[(trichloromethyl)thio]-H-isoindole-1,3(2H)-dione phthalimide(I))     

五唇兰种子离体培养的研究

授粉90d的五唇兰种子95%具球形胚,无菌条件下播种于培养基3个月后种子最高萌发率达到 90%。ABA和高浓度的NH4NO3抑制种子的萌发。NAA和BA促进种子的萌发,最适浓度为BA0.2mg· L 1+NAA0.5mg·L 1。400mg·L 1的谷氨酰胺可促进原球茎生长。2000mg·L 1的蛋白胨促进原球茎发 育成苗。     

In vitro asymbiotic germination and seedling development of Limodorum abortivum (Orchidaceae)

Abstract

Germination tests were carried out using immature seeds of Limodorum abortivum and applying in vitro techniques. The results proved that BM₁ culture medium is suitable to promote both germination and further growth stages. Details of the developmental pattern, and some micromorphological features, are described.     

In vitro propagation of prairie gentian

Explants of shoot tips, internodal stem sections, and leaf segments of Lisianthus, Eustoma grandiflorum (Griseb.) Schinners, Dwarf Purple were cultured in vitro on modified Murashige and Skoog (MS) media. Explants of shoot tips and internodal stem sections developed into multiple shoots, whereas, leaf segments turned chlorotic on a medium supplemented with 3 mgl^-1 benzyladenine (BA) and 0.2 mgl^-1 naphthalene acetic acid (NAA). Shoot proliferation was obtained on shoot tips and leaf segments with 3 mgl^-1 BA, but internodal stem sections became necrotic and died on this medium. Rooting was induced in cultures with multiple shoots by subculturing explants on a half-strength MS medium supplemented with 2 mgl^-1 indole-3-acetic acid (IAA). Rooted plantlets were successfully transferred to soil.     

In vitro propagation of mature Liquidambar styraciflua

Mature specimens of liquidambar styraciflua were propagated in vitro. Components of the nutrient medium and culture conditions were first determined for one-year-old seedling material. Mature material responded similarly to seedling material in culture, but alterations in frequency of early transfers and components of the medium were required. Explants responded best to Woody Plant Medium of Lloyd and McCown supplemented with 0.2 mg l^-1 BA and 0.05 mg l^-1 NAA. Root formation occurred on shoots placed on media containing 0.5–1.0 mg l^-1 IBA. Growth in culture and percentage of rooting of mature explants were markedly affected by the individual selection, with rooting percentages varying from 33–100% among selections.     

In vitro propagation of Pelecyphora aselliformis Ehrenberg and P. strobiliformis Werdermann (cactaceae)

Summary Development of efficient in vitro propagation, systems for Pelecyphora aselliformis Ehrenberg and P. strobiliformis Werdermann, two endangered Mexican species of cacti, are described. Multiple shoot formation from areoles of in vitro-germinated plantlets was achived in two types of explants (apical and transversal) cultured in Murashige and Skoog (MS) basal media supplemented with 30 or 50 gl⁻¹ sucrose, 10 gl⁻¹ agar and various treatments with cytokins. Shoot production in these proliferation media was evaluated after one (60 d) and three (180 d) culture cycles. In P. asellifornis 13.7 shoots per explant were produced after the first cycle using apical explants in medium with 8.8 μM 6-benzylaminopurine (BA) and 30 gl⁻¹ sucrose. In P. strobiliformis the highest proliferation rate (12.4 shoots per explant) was reached using 8.8 μM Ba and 50 gl⁻¹ sucrose with shoot transverse segments as explants. After the third proliferation cycle, 128.1 and 136.3 shoots per explant were obtained in P. aselliformis and P. strobiliformis, respectively. The shoots were clongated in MS basal medium with 3 gl⁻¹ activated charcoal and rooted in MS basal media indoleacetic acid (2.85 or 5.71 μM) or indolebutyric acid (2.46 or 4.90 μM). On averge, rooting efficiency was 89% for P. aselliformis and 87% for P. strobiliformis. The survival frequency, of the plants once transferred, to soil was on average 88%.     

In vitro propagation of Dipterocarpus alatus and Dipterocarpus intricatus

Seedlings were grown in vitro from embryos of Dipterocarpus alatus and D. intricatus. The problem of explant browning could be overcome by growing embryos initially on a filter paper bridge in liquid medium with activated charcoal. The best basal medium was Woody Plant Medium without the ammonium nitrate. Cytokinin appeared to stimulate seedling growth, 5×10^-5 M 2-isopentenyladenine and 10^-4 M 6-benzyladenine (BA) being the optimum concentrations for D. alatus and D. intricatus respectively. Cotyledonary nodes, excised from the seedlings, were induced to form axillary shoots and in the case of D. intricatus these could be multiplied rapidly. D. intricatus shoots elongated by reducing the BA level from 10^-5 M to 5×10^-7 M. Roots developed when shoots were dipped in 10^-3 M indolebutyric acid for two minutes and subsequently grown in liquid medium supported by a filter paper bridge.Abbreviations AC activated charcoal - BA 6-benzyladenine - 2iP 2-isopentenyladenine - IBA indolebutyric acid - MS Murashige & Skoog medium - PVP polyvinylpyrrolidone - PVPP polyvinylpolypyrrolidone - WPM Woody Plant Medium - 1/2 WPM Woody Plant Medium with half-strength macro salts - WPM (-NH₄NO₃) Woody Plant Medium without ammonium nitrate     

In vitro propagation of a semi-dwarfing cherry rootstock

A successful in vitro propagation system for the semi-dwarfing cherry rootstock Maxma-14 (Prunus avium L.) has been developed. Shoot tips and axillary buds were successfully established in vitro. Multiplication rate of about 6 was achieved over a 4-week period using Murashige and Skoog medium with 4.44 μM benzyladenine and 0.49 μM indole-3-butyric acid (IBA). Rooting occurred within 4 weeks on liquid and agar-gelled media containing 0.49 μM NAA or 0.49, 2.45 μM IBA. On liquid media, a maximum rooting efficiency of up to 100% was obtained. However, high concentrations of auxins delayed the time of root initiation for 3–5 days. Acclimatization was affected directly by rooting conditions. Survival was best when plantlets were transferred to pots after a short period of root emergence on rooting media. Multiplication medium was also important for successful acclimatization. Shoots transferred to rooting media from that with kinetin resulted in better acclimatization and survival than that derived from media with benzyladenine. Further, plantlets rooted on liquid media had better survival than that rooted on agar-gelled media. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro propagation of cauliflower,Brassica oleracea var. botrytis for hybrid seed production

Methods for obtaining heterotic F₁ and maintaining purebred lines for breeding of Brassica oleracea are limited by absence of male sterile lines and occurrence of inbreeding depression, respectively. The use of vegetative (stem, petiole, leaf, leaf rib) and floral (peduncle, pedicel, flower bud, curd) explants of cauliflower to regenerate purebred lines for crossing were examined. Of four growth regulator treatments and explant types used, best results were obtained with curd explants on MS medium with 6-benzyladenine (cytokinin) and gibberellic acid. Although 6-benzyladenine alone promoted formation of shoots in floral explants, both 6-benzyladenine and α-napthaleneacetic acid were required for vegetative explants. Use of α-napthaleneacetic acid, however, often increased callus formation. These culture techniques to maintain purebred regenerated plants will complement newly-derived nuclear-based male sterile lines obtained by the introduction of antisense copies of the gene BcpI, which is required for pollen fertility. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro propagation of Agave fourcroydes Lem. (Henequen)

In vitro plant regeneration of Agave fourcroydes Lem. (Agavaceae) is described. Results suggest that the NO₃ ^-:NH₄ ⁺ balance in the culture medium is a key factor controlling callus growth and organogenesis in rhizome cultures. Stem callus showed limited organogenic capacity, but high cytokinin concentrations induced adventitious shoot formation on stem explants. When these shoots were excised and subcultured, new callus formed at their base from which new shoots arose. The shoots from stem explants and rhizome callus formed extensive root systems in vitro and were transferred to pot culture with a 90% survival rate.     

In vitro propagation of Halesia carolina L. and the influence of explantation timing on initial shoot proliferation

Halesia carolina L., a small, ornamentally valuable tree, is difficult to propagate due to the complexity of seed propagation and the unavailability of propagules for conventional vegetative propagation. A micropropagation system was developed to facilitate easy propagation of this species. Actively growing shoot tips achieved optimum shoot proliferation from axillary buds when placed on Woody Plant Medium supplemented with 1.0 to 2.5 mg/l benzyladenine. The addition of 0.1 mg/l naphthaleneacetic acid had little effect on culture performance. Murashige and Skoog medium was incapable of supporting vigorous shoot proliferation. Non-sterile rooting conditions provided better rooting and subsequent plantlet growth, when compared to an in vitro rooting method. The seasonal fluctuations in the stock plant dramatically affected the shoot proliferating potential of the explants in vitro. Rapidly elongating shoots formed shoot proliferating cultures more slowly than explants taken either before or after the rapid elongation phase.     

In vitro propagation of cassava (Manihot esculenta Crantz)

A method is presented for the rapid in vitro propagation of cassava (Manihot esculenta Crantz). Nodal explants were induced to grow as multiple-shoot cultures on a medium containing 1.0 M 6-benzylamino purine (BAP), supplemented with 0.25 M -naphthaleneacetic acid (NAA). Nodes were removed from the shoots after three weeks of growth and subcultured on fresh culture medium. An average of 7.0 nodes were produced from each explanted node after three weeks in culture. Nodal explants were transferred to a medium containing 2.5 M indole-3-butyric acid (IBA) to improve root initiation on the developing plantlets. Plant establishment was possible upon transfer to soil. In vitro propagation offers enhanced rates of multiplication over more conventional methods of propagation. In addition, in vitro propagation facilitates the storage and international exchange of cassava germplasm.