干旱胁迫对高山离子芥试管苗抗氧化系统及其活性氧代谢的影响

该试验以高山离子芥试管苗(Chorispora bungeana)为试材,采用固液培养法,设置对照(不添加PEG-6000,CK),轻度干旱胁迫(5%PEG-6000)、中度干旱胁迫(20%PEG-6000)、重度干旱胁迫(40%PEG-6000)4个干旱处理水平,分析干旱胁迫对高山离子芥幼苗抗氧化系统、活性氧代谢等部分生理特征的影响,以揭示高山离子芥在干旱胁迫下的生理响应特征,为进一步探讨其对干旱环境的适应机制奠定基础。结果显示:(1)随着干旱胁迫程度的增加以及在各时间胁迫处理下,抗氧化酶SOD活性及可溶性糖含量显著升高,POD活性、丙二醛含量、CAT活性和APX活性均经历了先升后降的过程。(2)超氧阴离子(O-·2)的产生速率和过氧化氢(H2O2)的含量均显著升高;高山离子芥试管苗叶片相对电导率呈现出升-降-升的变化趋势。(3)相关分析结果显示,MDA与相对电导率、可溶性糖、SOD、APX、O-·2及H2O2呈极显著正相关关系,可溶性糖与SOD、POD、O-·2及H2O2呈极显著正相关关系;相对电导率以及保护酶系均与O-·2、H2O2呈极显著正相关关系。研究表明,高山离子芥具有较强的耐旱性,高山离子芥试管苗在响应干旱胁迫过程中,抗氧化酶系、活性氧代谢、脂质过氧化及渗透调节物等共同参与了高山离子芥试管苗对干旱胁迫的综合抗逆性形成,从而积极启动应对外界干旱环境的耐旱响应机制。     

Development of suitable protocol to overcome hyperhydricity in carnation during micropropagation

Hyperhydricity during micropropagation of carnation (Dianthus caryophyllusL.) was reduced by media modifications. Three commercial varieties (White sim, Exquisite and Scania) tested, varied in optimal growth without hyperhydricity. Increased concentration of iron and/or magnesium reduced hyperhydricity with 0.7–0.8% agar. At some concentrations, hyperhydricity was reduced to 0% and shoot multiplication was increased. All nonhyperhydrified micropropagated plantlets survived in a glasshouse during acclimatization.     

彩叶草对模拟干旱胁迫的生理响应

采用盆栽试验,对彩叶草进行PEG-6000浓度为0(对照)、5%、10%、15%、20%(W/V)模拟干旱胁迫,研究在干旱胁迫下彩叶草的生长、渗透调节能力及抗氧化酶活性的变化。结果表明:与对照相比,随着PEG-6000浓度的增加,鲜质量、干质量、含水量、水势、根系脱氢酶活性、无机离子含量包括K+、Na+、Ca2+、Mg2+等均呈下降趋势;NO-3含量呈先下降后上升趋势;硝酸还原酶活性、可溶性蛋白含量、可溶性糖含量、超氧化物歧化酶活性均呈先上升后下降趋势;脯氨酸含量、游离氨基酸含量、过氧化物酶活性、过氧化氢酶活性、超氧阴离子(O-2·)产生速率、质膜透性则呈上升趋势。因此,模拟干旱胁迫对彩叶草生长有抑制作用,且随着PEG-6000浓度增加,其生长受抑制和水分胁迫程度加重;模拟干旱胁迫下,彩叶草不积累K+、Na+、Ca2+、Mg2+和NO-3等无机离子进行渗透调节,而积累脯氨酸、可溶性蛋白、可溶性糖、游离氨基酸等有机小分子物质进行渗透调节,但这4种小分子物质增加幅度不尽相同;轻度模拟干旱胁迫虽增强彩叶草抗氧化酶活性,但仍表现轻度的氧化伤害;重度模拟干旱胁迫加重彩叶草氧化伤害。研究结果可为彩叶草耐旱生理机制的研究积累资料,也为其节水型栽植和养护提供依据。     

AgNO<Subscript>3</Subscript> prevents the occurrence of hyperhydricity in <Emphasis Type="Italic">Dianthus chinensis</Emphasis> L. by enhancing water loss and antioxidant capacity

Hyperhydricity symptoms are common and significant during the in vitro culture of Dianthus chinensis L. and greatly affect the micropropagation and regeneration of cultured plantlets. However, effective measures for preventing such abnormalities have not been developed for this species. Silver nitrate (AgNO₃) has been shown to revert hyperhydric plantlets to a normal state. Nevertheless, the effect of AgNO₃ on the prevention of hyperhydricity and the underlying mechanisms remain unclear. In the present study, 98.7% of the Dianthus chinensis L. plantlets cultured in a hyperhydricity induction medium (HIM) developed symptoms of hyperhydricity; however, hyperhydricity symptoms were inhibited to different degrees when D. chinensis L. plantlets were cultured in HIM supplemented with various concentrations of AgNO₃. In particular, approximately 97% of the D. chinensis L. plantlets grew normally and did not show any symptoms of hyperhydricity when cultured in HIM supplemented with 30 μmol L^?1 AgNO₃. Compared with the plantlets cultured in HIM alone, the plantlets cultured in HIM containing AgNO₃ displayed dramatic decreases in water content, ethylene content, and reactive oxygen species (ROS) production (particularly regarding H₂O₂ accumulation in guard cells) and showed increased antioxidant enzyme activity, stoma aperture, and water loss. These changes not only prevented excess water from accumulating in the tissues of plantlets but also improved the antioxidant capacity of plantlets, ultimately resulting in the prevention of hyperhydricity.     

Analysis of ultrastructure and reactive oxygen species of hyperhydric garlic (Allium sativum L.) shoots

Hyperhydricity is a physiological disorder frequently affecting shoots propagated in vitro. Since it negatively affects shoot multiplication vigor, and impedes the successful transfer of micropropagated plants to in vivo conditions, hyperhydricity is a major problem in plant tissue culture. In commercial plant micropropagation, there are reports of up to 60% of cultured shoots or plantlets which demonstrate hyperhydricity, which reflects the pervasiveness of this problem. The phenomenon has been correlated to water availability, microelements, and/or hormonal imbalance in the tissue culture. In this study, the ultrastructure and the characteristics of reactive oxygen species between hyperhydric and normal shoots of garlic were studied. We observed that in some cells of hyperhydric tissues, the intranuclear inclusion was separated, the mitochondrion was swollen and its intracristae had splits, the organelles were compressed against the cell wall, and the chloroplasts and intergranal thylakoids were also compressed. Additionally, the content of chlorophyll and soluble protein in hyperhydric shoots decreased significantly. For instance, chlorophyll a decreased 43.61%, chlorophyll b decreased 49.29%, chlorophyll a+b decreased 48.10%, and soluble protein dropped 47.36%. In contrast, the O₂ generation rate and H₂O₂ level increased 45.36% and 63.98%, respectively, obviously higher than the normal shoots. Lipoxygenase activity and malondialdehyde content in the hyperhydric shoots increased significantly, while the electrolyte leakage rose, indicating a serious membrane lipid peroxidatic reaction. Superoxide dismutase, peroxidase, catalase, glutathione peroxidase, and ascorbate peroxidase activities in hyperhydric tissue were all significantly higher than in normal leaf tissue. The antioxidant metabolism demostrated a close connection between hyperhydricity and reactivated oxygen species.     

Isolation and Pathogenicity of Rhizosphere Fungi of Cocoyam in Relation to Cocoyam Root Rot Disease

Cocoyam is the second most important staple crop of Cameroon and root rot is a destructive disease of this plant. Pythium myriotylum (Pm), Fusarium solani (Fs), and Rhizoctonia solani (Rs) were isolated from the rhizosphere of root rot affected cocoyams and from the soil of a cocoyam experimental field plot temporarily devoid of same in Mamu, Cameroon. Pm was isolated from the above soil by the cocoyam leaf disc baits. Fs and Rs were also isolated from the same soils by the water dilution method and from the roots of diseased cocoyams but were always associated with mycelial growth of Pm. Pathogenicity of Pm and in combinations with Fs or Rs or Fs + Rs all developed cocoyam root rot disease (CRRD) symptoms on 3– and 7–month old cocoyam plantlets 2–7 days after inoculation. Symptoms included rotted roots and wilting with general chlorosis of inoculated plantlets. No symptoms of CRRD were noted on cocoyam plantlets inoculated with Fs, Rs, Fs + Rs, and distilled water. Results indicated that CRRD is not caused by several pathogens but only by Pm. Pm isolates from the soils and roots of diseased cocoyams and those maintained in the ROTREP laboratory have significantly bigger diameter of mycelial colony growth in 24 h–period at 31 °C on lima bean sucrose agar, V–8 juice sucrose agar, and potato sucrose agar than on potato dextrose agar and 2 % water agar. The cocoyam plantlets were raised axenically from tissue culture of explants in the laboratory.     

Effect of PEG-6000 imposed drought stress on RNA content,relative water content (RWC), and chlorophyll content in peanut leaves and roots

Drought, one of the environmental stresses, plays crucial role in reduction in plant production on majority of agricultural fields of world, In order to evaluate drought stress on RNA content Relative water content (RWC), and chlorophyll content, Water deficit was induced by Polyethylene glycol (PEG) in peanut (Arachis hypogaea), accession number ICGV 91114. In this current study we evaluate RNA content and Relative water content (RWC) both in leaves and roots and chlorophyll content in leaf. The present study was undertaken with the aim to investigate the effect of water deficit imposed by PEG-6000, 40 old day seedlings were treated with varying concentrations of polyethylene glycol-6000 (PEG-6000; w/v-5%, 10%, 15% & 20%) for 24 h. The results showed that RNA content and Relative water content (RWC) content was significantly reduced in both leaves and roots with increased concentration of PEG, In leaves, a concentration dependent decline in chlorophyll content with increasing concentration of polyethylene glycol-6000 (PEG-6000). Reduction in chlorophyll ‘a’ level was to a greater extent than the chlorophyll ‘b’. Thus, this attributes can be used as screening tool for drought tolerance in peanut.     

水杨酸对高温胁迫下铁皮石斛幼苗耐热性的影响

为研究不同浓度水杨酸对铁皮石斛幼苗耐热性的诱导效应,以移栽半年的铁皮石斛幼苗为实验材料,对不同浓度水杨酸诱导高温胁迫下铁皮石斛幼苗的耐热性进行外观评价及叶绿素、可溶性蛋白质、可溶性糖、丙二醛(MDA)、游离脯氨酸(Pro)含量以及超氧化物歧化酶(SOD)、过氧化酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性的动态测定,观察其动态变化趋势并筛选最佳施用浓度.结果显示:在高温胁迫环境下,随着胁迫时间的延长,不同浓度SA处理均能有效缓解高温对铁皮石斛植株伤害;1.5～2.0 mmol·L1SA处理使铁皮石斛叶片中SOD、POD活性显著提高,0.5～1.5 mmol·L-1 SA处理叶片中CAT、APX活性显著提高;1.5～2.0 mmol·L-1SA处理可显著促进叶片Pro、可溶性糖和可溶性蛋白的积累,有效抑制MDA含量的增加,且不同浓度处理之间差异显著.研究表明,适宜浓度水杨酸处理能提高铁皮石斛幼苗的耐热性,并以1.5 mmol·L-1浓度处理效果最好.     

植物色素在24-表油菜素内酯调节油菜耐盐性中的作用

为了探索24表油菜素内酯(24-EBL)对盐胁迫下油菜生长的调节效应和植物色素在油菜耐盐性中的作用,采用盆栽实验,在盐胁迫下外源喷施1 000、10、0.1、0.001 nmol·L-1 24-EBL处理油菜幼苗,测定植株的生物量、电解质渗漏率(ELP)、净光合速率(Pn)、光合色素、酚类、类黄酮、花青素含量以及抗氧化能力(T-AOC).结果显示:(1) 24-EBL可显著缓解盐胁迫对油菜幼苗的氧化伤害,提高盐渍下油菜幼苗Pn和光合色素含量,并以0.1nmol·L-1 24 EBL(EBL3)对生长的调控效应最佳.(2)盐胁迫下,植株不同部位叶片的β-胡萝卜素(β-Car)和叶黄素(Lut)含量均显著下降,EBL3处理可显著提高其上部叶的β-Car含量,以及上部和中部叶的Lut含量;EBL3处理可显著提高盐胁迫下油菜所有叶片和叶柄的酚类含量,以及叶柄中类黄酮含量;EBL3处理可显著提高盐胁迫下油菜幼苗所有器官的花青素含量.(3) EBL3仅能够诱导上部叶和中部叶类胡萝卜素(Car)提取液的抗氧化能力(T-AOC)提高,但可诱导植株所有器官的酚类提取液的T-AOC提高.(4)不同部位的叶片Car、β-Car和Lut含量均与其Car提取液的T-AOC呈极显著正相关;而上部叶的总酚和花青素含量、中部叶和叶柄的花青素含量及茎秆中总酚、类黄酮和花青素含量与各自的酚类提取液的T-AOC呈极显著正相关.研究表明,外源喷施适宜浓度的24-EBL能够显著促进盐渍条件下油菜幼苗的光合能力,提高其抗氧化能力,从而增强其对盐渍胁迫的适应性,而光合色素和花青素水平被24-EBL诱导上升在油菜幼苗抗氧化过程中起着重要的作用.     

Agar and ammonium nitrate influence hyperhydricity,tissue nitrate and total nitrogen content of serviceberry (Amelanchier arborea) shoots in vitro

Shoot tip cultures of Amelanchier arborea Michx.f. were grown on Murashige & Skoog or Woody Plant (WP) medium containing 4.4 M benzyladenine and various concentrations of agar. Increases in agar concentration affected various culture growth variables, decreased culture hyperhydricity and increased tissue nitrate concentration. Additions of ammonium nitrate to cultures grown on WP medium containing 0.4% agar increased all growth variables measured except percent dry weight. Hyperhydricity and tissue nitrate concentration also increase in response to increasing ammonium nitrate in the medium. Since hyperhydricity was shown to be both positively and negatively correlated with increases in tissue nitrate content, it is unlikely that tissue nitrate level alone directly affects hyperhydricity.Abbreviations BA benzyladenine - MS Murashige & Skoog - WP Woody Plant     

Endogenous cytokinins in shoots of Aloe polyphylla cultured in vitro in relation to hyperhydricity, exogenous cytokinins and gelling agents

The process of hyperhydricity in tissue cultured plants of Aloe polyphylla is affected by both applied cytokinins (CKs) and the type of gelling agent used to solidify the medium. Shoots were grown on media with agar or gelrite and supplemented with different concentrations of N⁶-benzyladenine (BA) or zeatin (0, 5 and 15 μM). Endogenous CKs were measured in in vitro regenerants after an 8-weeks cycle to examine whether the hyperhydricity-inducing effect of exogenous CKs and gelling agents is associated with changes in the endogenous CK content. On media with agar a reduction in hyperhydricity occurred, while the gelrite treatment produced both normal and hyperhydric shoots (HS). The content of endogenous CKs, determined by HPLC-mass spectrometry, in the shoots grown on CK-free media comprised isopentenyladenine-, trans-zeatin- and cis-zeatin-type CKs. The application of exogenous CKs resulted in an increase in the CK content of the shoots. Following application of zeatin, dihydrozeatin-type CKs were also detected in the newly-formed shoots. Application of BA to the media led to a transition from isoprenoid CKs to aromatic CKs in the shoots. Shoots grown on gelrite media contained higher levels of endogenous CKs compared to those on agar media. Total CK content of HS was higher than that of normal shoots grown on the same medium. We suggest that the ability of exogenous CKs and gelrite to induce hyperhydricity in shoots of Aloe polyphylla is at least partially due to up-regulation of endogenous CK levels. However, hyperhydricity is a multifactor process in which different factors intervene.     

Micropropagation of <Emphasis Type="Italic">Cotoneaster wilsonii</Emphasis> Nakai—a rare endemic ornamental plant

A simple and efficient micropropagation system was developed for Cotoneaster wilsonii through node and shoot tip explants obtained from mature field-grown plants. Of the two explants, node explants were found to be the most effective for axillary shoot proliferation. The highest frequency of shoot induction was achieved when nodal explants were incubated on Murashige and Skoog (MS) medium supplemented with 0.5 mg L⁻¹ thidiazuron (TDZ) and 0.1 mg L⁻¹ α- naphthaleneacetic acid (NAA) with an average of 34 shoots per explant. The microshoots were separated from the multiple shoots and subcultured on MS medium supplemented with 3% (w/v) sucrose and 0.8% (w/v) agar for further shoot growth. Maximum rooting was obtained on half-strength MS medium supplemented with 0.5 mg L⁻¹ indole-3-butyric acid (IBA). The in vitro-grown plantlets were successfully acclimatized in a glasshouse with 98% of survival. High concentrations of TDZ (1.5–2.0 mg L⁻¹) and repeated subcultures resulted hyperhydric shoots. Supplementation of the culture medium with silicon significantly reduced the induction of hyperhydric shoots. Increasing silicon concentration significantly decreased malondialdehyde content of the regenerated shoots. Data indicate that addition of silicon to the culture medium can effectively control hyperhydricity.     

Somatic embryogenesis and plantlet production using rejuvenated tissues from serial grafting of a mature <Emphasis Type="Italic">Kalopanax septemlobus</Emphasis> tree

An effective micropropagation technique via somatic embryogenesis has been developed using tissue from serially grafted shoots generated from a mature Kalopanax septemlobus tree (~40 y old). Callus was induced from leaf segments obtained from the grafts by culturing the explants in Murashige and Skoog (MS) medium supplemented with 2,4-D and 3% w/v sucrose under darkness. The effects of sucrose, coconut water, and polyethylene glycol (PEG-3350) were evaluated as factors to promote development of somatic embryos (SEs) from embryogenic callus. More than 90% of explants formed callus; however, only 2.5%, or 20 leaf segments out of 800 explants, formed embryogenic callus after 8 wk of culture. High sucrose concentrations (3% and 5% w/v) were effective in inducing SEs. Treatment with 2–10% v/v coconut water also had a positive effect on embryo induction. A synergistic effect on SE induction was obtained using sucrose and PEG, with presence of the latter compound resulting in smaller, more uniform SEs. Embryo germination and conversion to plantlets were significantly influenced by the gelling agents. In general, gelrite-gelled medium was superior to agar-gelled medium. In gelrite-gelled medium, gibberillic acid (GA₃) enhanced embryo germination. Converted plantlets in an artificial soil mixture showed a 91% survival rate and displayed no distinct morphological variations. Our results indicate that reliable somatic embryogenesis and plant production can be achieved with rejuvenated tissues after repeated grafting of shoots derived from a mature Kalopanax septemlobus tree.     

水分胁迫对白刺幼苗生物量和渗透调节物质积累的影响

以PEG-6000模拟不同程度的水分胁迫对白刺幼苗进行处理,研究了其植株干重及其K 、Na 、丙二醛、游离脯氨酸、可溶性糖的含量变化.结果表明,15%PEG胁迫下白刺幼苗的生物量最高,丙二醛含量最低,且二者与对照的差异均显著;随着水分胁迫程度的增强,K 含量逐渐降低并与对照差异显著,Na 含量先减少后增加而其总积累量无显著变化;游离脯氨酸和可溶性糖含量随着水分胁迫程度的增强而显著增加.因此,轻度水分胁迫有利于白刺幼苗的生长,有机溶质游离脯氨酸和可溶性糖是白刺适应干旱环境的主要渗透调节物质.     

硝酸镧和兰科菌根真菌对铁皮石斛生理特性的影响

利用盆栽的方式研究了不同硝酸镧水平(0、1.0、3.0、5.0和7.0mg·L-1)下接种兰科菌根真菌对铁皮石斛生物量、多糖和蛋白质合成的影响,并分析了叶绿素含量、丙二醛(MDA)含量以及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)活性的变化,以探讨硝酸镧和兰科菌根真菌对铁皮石斛生理特性的影响。结果表明:(1)添加适量的硝酸镧有利于菌根真菌侵染和菌根发育,提高铁皮石斛幼苗生物量。(2)在接种菌根真菌的同时添加5.0mg·L-1的硝酸镧,铁皮石斛的根重、茎叶重和总生物量均达到最大,分别是未添加硝酸镧以及未接种对照组的4.26倍、4.98倍和4.87倍,其菌根侵染率也高达92.8%;而且可显著提高叶片中叶绿素含量,并显著降低细胞内的丙二醛含量。(3)在适量(5.0mg·L-1)的硝酸镧水平下接种菌根真菌能促进铁皮石斛幼苗多糖和蛋白质的合成,并显著提高细胞内SOD、CAT和POD活性。研究认为,菌根真菌与适宜浓度硝酸镧(5.0 mg·L-1)联合使用能显著促进铁皮石斛菌根的形成,增强植株的生理活性和适应能力,提高其生物量和多糖等活性成分的积累,有效改善铁皮石斛的药用品质。     

芦笋在PEG模拟干旱条件下的生理生化变化

用聚乙二醇(PEG-6000)胁迫处理芦笋幼苗,检测了幼苗的渗透调解物质含量、生物膜透性、抗氧化特性等指标。结果表明:脯氨酸、可溶性糖含量随着处理时间的延长显著增加,且以20%的PEG浓度处理增加最明显;SOD、POD、CAT活性在PEG处理后明显增强;相对电导率随着处理浓度的增大和时间的延长而增大;MDA含量随着处理时间的延长明显增加,且以10%和20%浓度处理增加显著。由此表明,芦笋在干旱胁迫下通过增加渗透调解物质含量,降低水势来提高其抗旱能力;通过增强抗氧化酶活性,提高抗氧化能力,来减轻干旱胁迫伤害。     

Promoting root induction and growth of in vitro macadamia (<Emphasis Type="Italic">Macadamia tetraphylla</Emphasis> L. ‘Keaau’) plantlets using CO<Subscript>2</Subscript>-enriched photoautotrophic conditions

In this study, a rooting protocol was developed for macadamia plantlets with healthy roots and enhanced growth performance, along with enhanced photosynthetic capability. In vitro-grown shoots rooted in vented vessels containing vermiculite as the supporting material exhibited 100% frequency of root induction, whereas when shoots were grown in non-vented vessels containing a solidified Murashige and Skoog (MS) medium, the frequency of root induction was less than 30%. The formation of root with callus, hyperhydricity, and leaf necrosis was observed in this photomixotrophic closed system. The modification of the vented photoautotrophic system with different concentrations of CO₂ and sucrose were investigated using vermiculite as the supporter. The number of roots, root length, root surface area, fresh weight, and dry weight were significantly higher in plantlets grown in CO₂-enriched (1,000 μmol CO₂ mol⁻¹) photoautotrophic conditions. The water content in both root and shoot tissues of plantlets cultured under photoautotrophic conditions was maximized. In addition, shoot and leaf performances were enhanced in plantlets cultured under CO₂-enriched photoautotrophic conditions. The supplementation of sucrose (29–88 mM) to culture media in both ambient and elevated CO₂ conditions affected a reduction in the shoot and root performance of in vitro plantlets. Chlorophyll a, chlorophyll b, and total carotenoids in the leaf tissues of plantlets acclimatized in CO₂-enriched photoautotrophic conditions were enriched, leading to increasing photosynthetic abilities, including chlorophyll fluorescence and net photosynthetic rate. From this investigation, a root induction protocol was established and the production of healthy macadamia plantlets was successfully implemented using CO₂-enriched photoautotrophic conditions.     

旱盐互作对冬小麦幼苗生长及其抗逆生理特性的影响

采用水培方法,以不同浓度的PEG-6000（0、8.3%、12.6%（W/V））和NaCl（0、25、50 mmol·L^-1）溶液模拟不同程度的干旱胁迫及盐胁迫,研究了盐分对干旱胁迫条件下冬小麦沧-6001幼苗生长及其抗逆生理特性的影响.结果表明：在8.3%或12.6% PEG-6000处理条件下,添加25 mmol·L^-1NaCl均使植株干物质积累和植株含水量比单一PEG处理增加,同时叶片可溶性糖和可溶性蛋白质含量增加,丙二醛和脯氨酸含量下降,植株各部位Na⁺含量升高、K⁺含量下降;在12.6% PEG-6000处理条件下,添加50 mmol·L^-1NaCl对植株的胁迫效应高于单一PEG处理.表明在干旱胁迫条件下,加入适量盐分可缓解干旱胁迫对冬小麦幼苗生长的抑制.     

Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity in in vitro pear cultivar shoots

This study was made to clarify the effects of cytokinin type and their concentrations on shoot proliferation and hyperhydricity in in vitro Pyrus pyrifolia N. (`Hosui' and `Kosui') shoots. The shoots were subcultured in a woody plant medium supplemented with 0.5 M 3-indolyl-butyric acid, 3% (w/v) sorbitol, 0.8% (w/v) agar, and with cytokinins kinetin, 6-benzylaminopurine (BA), N-(2-chloro-4-pyridyl)-N9-phenylurea (CPPU), 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ) added at concentrations 0.44, 4.40, 11.0 and 44.0 M. The highest number of shoots was induced by adding BA at concentration 11.0 M, then by 4.4 M BA, in both cultivars. TDZ and CPPU caused greater hyperhydricity in cultured explants than BA and kinetin. `Kosui' was more susceptible to hyperhydricity compared with `Hosui'.     

Effects of sucrose concentration,supporting material and number of air exchanges of the vessel on the growth of in vitro coffee plantlets

Growth of coffee (Coffea arabusta) plantlets cultured in vitroas affected by sugar, types of supporting material and number of air exchanges of the vessel was investigated. Single node cuttings of in vitro coffee plantlets were cultured on half strength MS medium with or without 20 g l⁻¹ sucrose. Two types of supporting material, agar and Florialite, and two levels of air exchange expressed by number of air exchanges per vessel, 0.2 and 2.3 h⁻¹, were studied. At the end of a 40-day culture period, fresh weight, shoot length, root length and leaf area of plantlets when cultured on Florialite soaked in sugar-free medium and under the higher number of air exchanges were greater than those in sugar containing medium. Callus was observed at the shoot base of plantlets grown on agar medium containing sucrose. Photosynthetic ability of coffee plantlets in vitro was also significantly increased when grown on sugar-free medium with the high number of air exchanges and Florialite as a supporting material. This revised version was published online in June 2006 with corrections to the Cover Date.