In vitro flowering of green and albino Dendrocalamus latiflorus

To propagate Dendrocalamus latiflorus, we used in vivo inflorescences to produce calli on Murashige and Skoog basal (MS) medium supplemented with 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg/l kinetin, 250 mg/l polyvinyl pyrrolidone (PVP), and 1% coconut milk. Multiple shoots were generated on MS medium supplemented with 0.1 mg/l thidiazuron (TDZ). The green plantlets were successfully transferred to soil. Multiple albino shoots also regenerated and were able to proliferate on medium containing cytokinins, especially TDZ. Albino multiple shoots rooted in medium containing α-naphthaleneacetic acid (NAA), and callus formation was observed in the presence of 2,4-D and picloram. Green and albino regenerates flowered after 8 months of subculture. The flowering ratio increased to 44% after three treatments in medium containing 1 mg/l TDZ. Morphological observations revealed that the in vitro green and albino flower organs were normal. However, pollen derived from the in vitro flowers of both the green and albino plants were sterile.     

Organogenesis and terpenoid synthesis in Mentha arvensis

Leaf discs obtained from field grown plants of Mentha arvensis were used to initiate multiple shoots on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (5 mg l(-1)) and naphthaleneacetic acid (0.5 mg l(-1)). Profuse rooting was achieved when the well-grown shoots were cultured on half strength MS medium supplemented with indole-3-acetic acid (2 mg l(-1)). The regenerated plantlets were hardened and successfully transferred to soil and grown to maturity. Tissues at different stages of differentiation were analyzed for their essential oil content and characteristic monoterpene pattern. Tissue culture raised plants show the same essential oil profile as that of the parent plant. However, tissues at early stages of growth show distinct changes in oil composition, such as high levels of pulegone in shoot cultures.     

“102”杨茎尖培养与植株再生

“102”杨[(Populus tomentosa×P.bolleana)×P.tomentosa]为优良杂种杨。用常规无性繁殖方法育苗均未成功。自1986年起试用茎尖组织培养,现已获得一批再生植株,并移植大田成苗。试验以Ms培养基为基础,分别摸索出适于“102”杨茎尖组织培养的诱导培养基(附加激素为6—BA0.3,1.0mg/l或KT0.3,1.0。Mg/l分别与NAA0.05mg/l配伍)、芽增殖培养基(附加激素6—BA0.3mg/l)和生根培养基(用1/2Ms培养基,蔗糖15g/l,附加NAA0.02mg/l)。     

In vitro plantlet regeneration from Australian Red Cedar (Toona ciliata,Meliaceae)

In vitro regeneration of complete plants from nodal single-bud segments of 2-year-old Australian Cedar (Toona ciliata) trees were obtained under defined nutritional and environmental conditions. Explants were dissected from plants obtained by germination of seeds and growth in pots in a greenhouse. The best medium for shoot regeneration was that of Murashige and Skoog at 1/4 strength with 3% sucrose (1/4 MS), supplemented with 0.1 mg/l IBA and 0.5 mg/l BAP. Rooting of regenerated shoots was observed in MS medium with 0.1 mg/l IBA. Using mature tree material was more difficult. Forced flushing was used to induce shoot development on branches of a 10-year-old tree. Nodal segments of these epicormic shoots formed shoots in vitro on 1/4 MS + 0.01 mg/l IBA + 5 mg/l BAP, but rooting was never observed.     

Plantlet regeneration from leaf protoplasts ofPopulus euphratica

Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the cell density of 9×10⁴/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of 2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and plantlets were regenerated on 1/2 MS medium without a hormone. A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995).     

MICROPROPAGATION IN TISSUE CULTURE OFDAHURIAN LARCH

Dormant buds of Larix gmelinii(4-30years old)were cultured in vitro.Axillary budsgrew on the explants,and60％-65％ of the explants’axillary buds,a differentiation rate of 60％ wasobtained on the explants collected from the 30-year-old trees.The maximum number of axillarybuds was 26 in one induction per initial explant.Bud clusters were separated into individual budsand most of them elongated into shoots.A few roots grew on the shoots.The MS(Murashige andskoog)and SH(Schenk and Hildebrandt)were more efficient media than the WPM.(Woody PlantMedium).The best hormone Combinations for the axillary bud inductions were BA1+NAA0.01 andBA2+NAA0.2(mg/L).The procedure was as follows:(1)Apical buds were explanted on theno—hormone basal agar medium and grown for 1 or 2 weeks;(2)Explants were transferred onto thebud—inducing medium and grown for 2 weeks and then(3)Cultured on the basal medium withouthormones for axillary bud elongation;(4)Bud clusters were separated and cultured continuously toa minimum height of1     

Preservation of Juniperus thurifera L.:a rare endangered species in Algeria through in vitro regeneration

Juniperus thurifera L.is an endemic Cupres saceae from the Aure`s Mountains of north eastern Algeria and endangered,in part,due to the scarcity of viable seeds It is threatened by other abiotic factors and the lack of an effective management strategy will increase its risk o extinction.The dearth of information on its in vitro regeneration impedes its application in forest managemen programs.We therefore developed a micropropagation protocol using microcuttings with auxiliary buds.Cuttings were grown on different combinations of media supplemented with plant growth regulators at different concentrations.The highest number of shoots and branches regenerated from original shoots was obtained on Woody Plant Medium(WPM)supplemented with 6-benzylaminopurine(BAP)(0.5 mg L^-1)and 2,4-dichlorophe noxyacetic acid(2,4-D)(0.25 mg L^-1).The best elongation of shoots was achieved with WPM supplemented with0.5 mg L^-1of BAP and 0.25 or 1 mg L^-1 of 2,4-D.On the second subculture,shoots had a higher number of branches than those of the first.The highest rooting rate,38.8%,was obtained with shoots cultured in 1/2 Murashige and Skoog(MS)medium supplemented with 5.0 mg L^-1each of indol-3-butyric(IBA)and naphthalene acetic acid(NAA).Similarly,the highest root numbers and lengths were produced on 1/2 MS medium supplemented with IBA and NAA(5.0 mg L^-1each).During transfer to acclimatization,rates of plant losses of 50% occurred.The second part of the experiment showed that the best shoot callusing was on WPM supplemented with BAP and 2,4-D,with either the combination 0.5+0.25 or 0.25+0.25 mg L^-1.The results of this research provide a starting point for further studies on in vitro regeneration of J.thurifera for the sustainable management of its unique ecosystem in the Mediterranean basin.     

Shoot proliferation and callus regeneration from nodular buds of Drepanostachyum luodianense

This research reports on an efficient shoot proliferation and callus regeneration system for bamboo.Young, semi-lignified branches with one lateral bud from Drepanostachyum luodianense(Yi et R. S. Wang) Keng f.were used as explants. Disinfection with 0.1% HgCl_2 for 8 min was the optimum treatment and the best medium for bud initiation was Murashige and Skoog(MS) medium containing 3.0 mg L~(-1)6-benzyladenine(BA). Multiple shoots were induced from nodal shoot segments on MS medium containing 5.0 mg L~(-1) BA, 0.5 mg L~(-1) kinetin(Kin), and 1.0 mg L~(-1) naphthaleneacetic acid(NAA). The highest frequency of callus formation(65.6%) was on MS medium containing 4.0 mg L~(-1)2,4-dichlorophenoxyacetic acid(2, 4-D), 0.5 mg L~(-1) NAA, and 0.1 mg L~(-1) thidiazuron(TDZ). The optimum medium for callus proliferation was MS medium with 4 mg L~(-1)2,4-D, 0.5 mg L~(-1) TDZ and 0.5 mg L~(-1) NAA, and the optimum hormone combination was 4 mg L~(-1) BA ? 0.5 mg L~(-1) NAA for callus redifferentiation(up to 85.6%). A 100% rooting was achieved on MS medium supplemented with 2.0 mg L~(-1) NAA and 0.5 mg L~(-1)3-indole butyric acid(IBA). Rooted plantlets were acclimatized in a greenhouse in humus soil ? perlite(1:1) substrate. These micropropagated callus induction and regeneration systems for bamboo will be useful for genetic engineering and multiplication.     

An Efficient Micropropagation Protocol for High-Frequency Plantlet Regeneration from Liquid Culture of Nodal Tissues in a Medicinal Plant,Turnera ulmifolia L.

A highly efficient, stable, and cost-effective micropropagation protocol for the conservation of a medicinal plant Turnera ulmifolia L. was established from nodal tissues via multiple axillary shoot proliferation on using Murashige and Skoog’s (MS) liquid nutrient medium. To begin with, nodal explants were placed on agar gelled medium amended with 2.0 mg L^?1 6-benzylaminopurine (BAP) and 0.1 mg L^?1 indole-3 acetic acid (IAA) for shoot induction. Subsequently, elongation of regenerated shoots could be possible on liquid MS medium supplemented with 0.5 mg L^?1 BAP and Kin (kinetin) each along with 0.1 mg L^?1 IAA where high frequency of regeneration in terms of number of shoots (47.2 shoots/explant) was achieved. Furthermore, long and healthy shoots (4?5 cm in length) were rooted on agar gelled half-strength of MS medium supplemented with 2.0 mg L^?1 indole-3 butyric acid (IBA). Finally, in vitro regenerated plantlets were gradually acclimatized in the greenhouse and transferred to the field successfully.     

弗吉尼亚栎不定芽增殖及试管苗再生影响因子研究

[目的]探析弗吉尼亚栎组织培养中不定芽增殖和生根的关键影响因子,建立弗吉尼亚栎试管苗再生体系,为弗吉尼亚栎无性系选育提供技术支撑。[方法]以弗吉尼亚栎带芽茎段为外植体,采用不定芽增殖途径,研究弗吉尼亚栎组织培养过程中外植体选择、褐化控制以及培养基选择、激素配比对不定芽增殖和生根的影响。[结果]表明:向培养基中添加抗坏血酸、硫代硫酸钠和活性炭均显著降低外植体的褐化半径(p<0.05),其中,外植体在含有3.00、5.00 g·L-1活性碳的培养基中褐化半径最小。基本培养基筛选试验表明:以1/4MS或WPM为基本培养基时,平均芽长和芽数明显优于MS培养基。不定芽增殖最佳培养基为1/4MS+1.20 mg·L-1 6-BA,培养周期40 d,增殖倍数可达6.6。最佳生根培养基为1/4MS+0.50 mg·L-1 IBA+0.50 mg·L-1NAA,生根率达53.33%,且根粗壮,木质化程度较高。组培苗移植到灭菌河沙中,平均成活率达57.78%。[结论]培养基成分和激素配比是影响弗吉尼亚栎不定芽增殖和试管苗再生的主要因子,低盐培养基(1/4MS或WPM)不仅可以促进不定芽增殖,且能够减...     

Establishment of a high-frequency regeneration system in <Emphasis Type="Italic">Cerasus humilis</Emphasis>, an important economic shrub

Cerasus humilis is a species of small, perennial, drought-resistant and multipurpose deciduous shrub grown in arid and semi-arid conditions in northern China. In this study, an efficient protocol for the rapid micropropagation of C. humilis has been standardized using stem and/or leaf explants. Direct multiple shoot induction was observed when the stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. The highest shoot induction was obtained when stem explants from adult trees were cultured on MS medium supplemented with 2.0 mg L^?1 6-benzyladenine (6-BA) and 0.9 mg L^?1 α-naphthaleneacetic acid (NAA). The leaf and stem explants cultured on MS medium with 1.0 mg L^?1 6-BA and 0.6 mg L^?1 NAA, and 0.5 mg L^?1 6-BA and 0.8 mg L^?1 NAA, respectively, produced the highest induction frequency of callus. Maximum proliferation of callus was observed on MS medium containing a combination of 0.5 mg L^?1 6-BA with 0.6 mg L^?1 2,4-dichlorophenoxyacetic acid (2,4-d). Optimal shoots differentiated from callus were obtained on MS medium supplemented with 5.0mg L^?1 6-BA and 0.9 mg L^?1 NAA. In vitro rooting was achieved on half-strength (1/2) MS medium containing 0.5 mg L^?1 NAA. Rooted plantlets were hardened under control conditions and successfully acclimatized under field conditions.     

孔雀草杂交F1代的植株再生

以孔雀草（Tagetes patula）子叶、下胚轴和叶片为外植体,通过器官直接发生途径诱导形成不定芽,探讨植物生长调节剂组合、AgNO3浓度、蔗糖浓度和外植体类型等因素对植株再生的影响,建立了再生体系。结果表明：MS＋6-BA 1.0 · L^- 1＋NAA 0.5 · L^- 1＋AgNO31.0 · L^- 1＋蔗糖40 g·L^-1培养基最适合不定芽的分化和增殖,子叶不定芽分化率达90%以上,平均每外植体分化不定芽数达5.3个。不定芽较适生根培养基为1/2MS＋IAA 0.2 · L^- 1＋NAA 0.1 · L^- 1,生根率达到90%。     

Effect ofl-phenylalanine on rooting of shoots from axillary buds of adultLarix leptolepis

Rooting of shoots derived from axillary buds was examined to establish an efficient shoot culture system of clonal micropropagation in adult tree ofLarix leptolepis Gord. (Japanese larch). Nine out of ten shoots induced calli (90%) on their shoot bases, and the two of them formed root primordia with a red pigment (20%) on the calli surface within 5 weeks after culturing on modified Murashige and Skoog (MS) medium supplemented with 1.5 μM of indolebutyric acid (IBA) and 1.5 μM of naphthaleneacetic acid (NAA). However, the primordia did not elongate actively. The addition of 10 mMl-phenylalanine in the MS medium with the auxins resulted in the formation of roots at high frequency, about 80%, and they elongated actively. Although callus was formed in all the shoots cultured on the medium withl-phenylalanine, it appeared that the callus development was less as compared to the medium withoutl-phenylalanine. Consequently, the rooting might be associated with the suppression of the induced callus.     

火炬松丛生芽诱导及植株再生

采用火炬松带子叶项芽为外植体建立了组织培养再生体系。结果表明,火炬松丛生芽的形成受诱导培养基中激素浓度及外植体年龄的影响较大。适合火炬松丛生芽诱导的培养基为改良GD 6-BA 4 ms／L NAA 0．02 mg/L(诱导率86．7％),截取外植体的最佳苗龄为14-21 d。已分化的丛生芽继代培养在无6-BA但附加0．5- 1．0 g/L活性炭或0．02-0．05 mg/L NAA的培养基上伸长很快。伸长的丛生芽接种在附加0．05 ms／L NAA的1／2改良GD培养基中,1个月后有12．5％产生不定根。     

Rapid Micropropagation of Edible Bamboo Dendrocalamus asper

Abstract

Micropropagation protocols for Dendrocalamus asper using nodal shoots and seeds culture are described. Multiple shoots were induced through forced axillary branching. Ninety-five percent of the nodal shoot explants taken from juvenile primary and lateral branches, produced multiple shoots through axillary buds activation within 2 to 8 weeks on Murashige & Skoog's (MS) medium supplemented with 0.1-15 mg/l benzyladenine (BA). The cultured seeds also produced multiple shoots (1-20) within 6 weeks on this medium. The multiple shoot differentiation was influenced by the concentration of BA in the medium. The in vitro generated shoots were excised and subculture on MS + 3.0 mg/l BAP for further shoot multiplication. Fifteen to 20 fold rate of shoot multiplication was achieved by regular subculturing. These shoots were multiplied for more than 3 years without loss of vigor. Ninety-five percent of the shoots were rooted, when propagules (each consisting of cluster of 3 shoots) were transferred on to MS medium with 3.0 mg/l NAA or 10 mg/l indole-3-butyric acid (IBA).

To date, 18,000 plants (through axillary bud initiated from nodal ex-plants) and 6,000 plants from seed culture have been hardened and acclimatized. 12,000 plants have been field transferred.     

大花萱草新品种“御衣黄”组织培养及植株再生

以大花萱草“御衣黄”花蕾为外植体建立其组织培养再生体系.研究结果表明,大花萱草“御衣黄”花蕾经75％乙醇30 s +0.1％升汞10 min处理后可获得较为理想的灭菌效果；诱导培养基MS+ 6-BA 5 mg/L+ NAA 5mg/L是花蕾理想的脱分化培养基,愈伤组织诱导率达到75％；在降低了激素质量浓度的培养基MS+...     

Shoot multiplication of Syringa reticulata var. mandshurica from in vitro cultured seedlings

We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog(MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine(BA) and indole-3-butyric acid(IBA). The better medium for shoot multiplication and growth was MS + 5 mg L~(-1)BA +0.5 mg L~(-1)IBA + 20 g L~(-1)sucrose +7 g L~(-1)agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium(macro-elements of MS medium are at half-strength) supplemented with 1 mg L~(-1)IBA, and the survival percentage was 80 % at 16 weeks after transplanting.     

Plant recovery from seedling-derived shoot tips ofFaidherbia albida grown in vitro

In vitro cultures were initiated from shoots taken from seedlings ofFaidherbia albida on Murashige and Skoog based medium supplemented with different combinations of 6-benzylaminopurine (BA) and -naphthaleneacetic acid (NAA). The shoots grew and rooted on all media. Rooting and vigorous growth were most successful on medium supplemented with 10^–7M NAA alone on which 87% of the shoots formed roots. Seventy-one percent of plantlets which were transferred to soil were successfully established and nodulated with the nativeRhizobium. The procedures provide a basis for the development of in vitro techniques for rapid multiplication and physiological studies of the species.     

Efficient method of micropropagation and in vitro rooting of teak (Tectona grandis L.) focusing on large-scale industrial plantations

Multiple shoots of high quality were produced in vitro from nodal expiants of Tectona grandis. An average of about 4 shoots/uninodal expiant was obtained within 4 weeks of culture on Murashige and Skoog’s (mMS) medium modified by 50% reduction in NH₄NO₃ concentration, supplemented with benzylaminopurine (1.5 mg L^?1); indole-3-butyric acid (0.01 mg L^?1) and gibberellic acid (0.1 mg L^?1). The latter was applied both in the medium and by soaking the nodal segments for 10 s. in a gibberellic acid solution of 100 mg L^?1. Hundred percent of shoots rooted cultured on modified MS medium containing IBA (0.5 mg L^?1) and putrescine (160 mg L^?1). Putrescine promoted both strong and highly ramified roots and fast growing shoots during the rooting phase, conditioning the plantlets for a good survival and quality. Plantlets were transferred to jiffy pots for a short acclimatization stage in greenhouse where they survived at 100%. This highly reproducible procedure can be adopted for large scale teak propagation.