Micropropagation and accumulation of essential oils in wild sage (Salvia fruticosa Mill.)

A protocol for in vitro propagation of the wild three-lobed sage (Salvia fruticosa Mill.) (Synonym, Salvia triloba L.) was developed. Shoot tips were excised from in vitro seedlings and established on MS, Nitch and Nitch (NN), or B5 medium. For shoot proliferation, in vitro nodal and apical explants were cultured on MS medium containing 0.25–2 μM 6-benzylaminopurine (BA), 6-furfurylaminopurine (kinetin), or thidiazuron (TDZ). Proliferated microshoots were rooted on MS medium supplemented with 2.7–11.4 μM indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), or α-naphthaleneacetic acid (NAA). Results indicated that shoots established at 100% regardless of media type, however, shoot height, nodes per shoot, and leaf number were highest for explants established on MS medium compared to NN or B5. Number and height of proliferated shoots, nodes per shoot, and leaf number were highest for nodal explants cultured on a medium containing 0.75 μM BA. Microshoots cultured on a medium supplemented with 2.7 μM IBA exhibited the highest rooting percentage compared to those cultured with IAA or NAA. Essential oil composition in microshoots and shoots of greenhouse-grown plants was determined by gas chromatography/mass spectrometry. The major essential oils detected in both plant materials were α-pinene, 1,8-cineole, camphor, and borneol. No α-thujone or β-thujone was detected. The content of essential oils, camphor, and borneol were higher in the microshoots than in shoots of greenhouse-grown plants.     

Significance of explant preparation and sizing in Aloe vera L.—A highly efficient method for in vitro multiple shoot induction

The present study was carried out to assess the effect of explant preparation and sizing for in vitro micropropagation of Aloe vera L. The stem nodal explants and shoot tips were cultured on modified Murashige and Skoog's medium (1962) supplemented with different concentrations of 6-benzylaminopurine (BA), kinetin (KIN), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and α-naphthaleneacetic acid (NAA) either singly or in combination. The best media composition was found to be MS medium supplemented with IAA (11.42 μM), IBA (9.8 μM) and BA (8.88 μM). The explants were divided into 2 sets, with and without ensheathing leaf base. Explant sizing, pruning and retention of mother tissue was highly significant in induction of multiple shoots and roots. The stem nodal explants with leaf base performed much better than those without such covering. A very high number of shoots and roots grew from these explants. The rooted plantlets were successfully acclimatized and transferred to the green house conditions and finally to field conditions.     

生长调节剂诱导西葫芦、黄瓜雌花分化和发育的研究

应用25、50、100和200mg／kg的吲哚乙酸（LAA）、萘乙（NAA）、赤霉素（GA）喷施2～3叶期西葫芦和黄瓜幼苗及用50、100和200mg／kg的TiBA（三碘苯甲酸）喷施20-30cm高的西葫芦和黄瓜植株,发现50mg／kgIAA和200mg／kgTiBA明显促进西葫芦雌花分化和坐果,NAA的促进效果较差,GA无效;50mg／kgIAA和100mg／kgTiBA促进黄瓜雌花分化和坐果,NAA的捉拼效果较差。低浓座的GA具有一定的促进作用。     

Factors affecting tissue culture of Damask rose (Rosa damascena Mill.)

The present study was conducted to evaluate the regeneration ability of Damask rose. Single-node explants were surface sterilised with 10% chlorox for 15 min and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of N⁶-benzyladenine (BA) or kinetin (Kin) separately or in combination with different concentrations of indole-3-butyric acid (IBA). Combination of 2.5–3 mg/l BA and 0.1 mg/l IBA was the most suitable treatment for proliferation. In vitro derived shoots were subcultured four times on the fresh medium within a 4-week period. Other treatments such as explant orientation (horizontal, vertical and oblique) and explant wounding were also examined but did not affect shoot multiplication rate significantly. Several experiments were carried out to stimulate in vitro rooting of Damask rose. Application of different media such as MS, 1/2 MS, 1/3 MS and 1/4 MS with different concentrations of indole-3-acetic acid (IAA), IBA and naphthaleneacetic acid (NAA) did not produce satisfactory results. Quick-dip method using sterilised 0–2000 mg/l IAA, IBA and NAA solutions was also studied. Other treatments such as using various concentrations of abscisic acid (ABA) in combination with various concentrations of IAA, IBA and NAA, and using various concentrations of sucrose and agar did not produce any roots. The best treatment for rooting of shoots was 2.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for 2 weeks in MS medium and then transferring the explants to MS medium without any growth regulator. Plantlets were acclimatised in a soil mixture consisting of peat moss and sand 1:1 (v/v) and successfully transferred to the greenhouse after 3 weeks.     

High frequency plant regeneration through adventitious multiple shoot organogenesis in epicotyl explants of Indian gooseberry (Emblica officinalis Gaertn)

Shoots regenerated adventitiously on epicotyl segments from in vitro seedlings of Emblica officinalis var. ‘Kanchan’. Epicotyls derived from 2-week-old aseptic seedlings were most responsive and produced a maximum number of 303 shoots per explant in Murashige and Skoog (1962) medium (MS) augmented with 8.8 μM N⁶-benzyladenine (BA) + 1.425 μM indole-3-acetic acid (IAA). Shoots readily elongated in MS lacking growth regulators and rooted in half-salt-strength MS (1/2 MS) supplemented with indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA). The highest rooting response was recorded in 1/2 MS containing 14.7 μM IBA. Plantlets were acclimatized inside the green house and 80% of the plantlets survived on transfer to garden soil.     

Black Currant Leaf Spot: II. Laboratory Tests of Fungicides for the Prevention of Sporing of Pseudopeziza Ribis on Overwintered Leaves

Summary

We have developed a two-step procedure for rooting of tea microshoots in vitro. The effectiveness of different auxin treatments for root formation was found to differ. Among the auxins tested, 25 μM -naphthalene acetic acid (NAA) gave the best results, with 100% rooting, compared to 25 μM indole-3-butyric acid (IBA) or 25 μM indole-3-acetic acid (IAA), which induced 17% and 58% rooting, respectively. Incubation of tea microshoots on 0.33 Murashige and Skoog (MS) medium, supplemented with 25.0 μM NAA or 175.0 μM IBA for 10 d, followed by transfer to auxin-free 0.33 MS medium resulted in 100% rooting, whereas 50.0 μM IAA induced 91.7% rooting. Besides the different auxin treatments, the strength of the MS medium, the duration of incubation of microshoots in auxin-containing medium, the sucrose concentration, the gelling agent, the pH of the medium, the incubation temperature, the light intensity, and the quality of the shoots also played a significant role during in vitro rooting of micropropagated tea shoots. Among the combinations tested, the most effective results were obtained when green microshoots were incubated on 0.33 MS medium supplemented with 25.0 μM NAA, 50.0 mM sucrose, pH 5.5, gelled with 0.2% (w/v) Phytagel^TM for 10 d at 25° – 30°C at a light intensity of 40 μmol m^–2 s^–1, followed by transfer of shoots to auxin-free 0.3 MS medium. This resulted in 100% rooting and, on average, 11 long roots were formed per shoot. Anatomical changes during adventitious rooting of micropropagated tea shoots in vitro were also studied to understand the process of rooting.     

磨盘柿离体叶片愈伤组织发生及不定芽诱导

研究了培养基种类、植物生长调节剂配比、叶片不同部位及培养条件对磨盘柿离体叶片愈伤组织发生及不定芽再生的影响。适宜磨盘柿叶片再生的基本培养基为改良MS(1／2 N),生长调节剂为ZT 4.0~6.0 mg/L或TDZ 1.0mg／L与IAA 0.1 mg／L组合,不宜用BA和高浓度的NAA;ZT和TDZ促进叶片再生的方式不同,前者利于直接再生,后者需降低细胞分裂素浓度,二次诱导使芽点生长成苗;叶片接种后暗培养2~3周再生效率高;叶片不同部位中以叶基部最易再生,其次为叶中部,叶尖最差。     

In vitro propagation of Blandfordia grandiflora (Liliaceae)

Multiple shoots were obtained when single shoot tips of Blandfordia grandiflora were exposed to different concentrations of kinetin, N6~ benzylaminopurine (BAP) and N6- isopentenylaminopurine (2iP). The best multiplication results were obtained when explants were subjected to kinetin at 8 (iM BAP at 0.5,2 and 8 (iM and 2iP at 2,8,32 and 128 |iM. Preliminary rooting trials were performed with three different auxins: indo- lebutyric acid (IB A), naphthalene acetic acid (NAA) and indole acetic acid (I A A). IB A at a concentration of 8 |iM and IAA at a concentration of 32 |xM gave highest root number, but when transplanted to the glasshouse, the best surviving plantlets were those treated with 2 |xM of IBA or 0.5 μM of IAA.     

Effect of gibberellic acid (GA3) on elongation and rooting of ‘St. Julien A’ rootstock in vitro

‘St. Julien A’ (Prunus instititia L.) rootstock was induced to proliferate shoots on a modified half-strength Murashige and Skoog (MS) medium. Cultures treated with 12.5 mg l^?1 gibberellic acid (GA₃) produced elongated shoots suitable for rooting. Elongated shoots were placed in media with indolebutyric acid (IBA) or indole-3-acetic acid (IAA) with or without a 16-day dark incubation. Light (16-h photoperiod) inhibited rooting. IAA (4 mg l^?1) was ineffective in promoting rooting. Rooting was best when shoots were incubated in the dark with IBA (4 mg l^?1). GA₃ was deleterious to shoots, causing chlorosis and apical die-back. Light regime interacted with auxin treatments in affecting shoot condition. Shoot condition was better on shoots treated with IBA and dark-incubated; while those treated with IAA were better when light-incubated.     

Effects of IAA and IAA precursors on the development,mineral nutrition,IAA content and free polyamine content of pepper plants cultivated in hydroponic conditions

The effect of IAA and two IAA precursors, l-tryptophan (Trp) and indole (Ind), on the growth, mineral nutrition and potential development under stress conditions of intact pepper plants cultivated in hydroponic conditions, has been studied.To this end, the growth of both shoots and roots, the plant content of both IAA and free polyamines and the content in leaves and roots of macronutrients and micronutrients have been evaluated.Both Trp and Ind presented a similar pattern of action to that of IAA regarding both plant development and mineral uptake. The application of Ind and Trp at the highest dose (10⁻³ M) was associated with significant increases of IAA content in roots. The application of IAA and IAA precursors was associated with an increase in Spermine and a decrease in Putrescine in leaves, thus suggesting the conversion of Putrescine into Spermidine and Spermine, which in turn could be associated with better plant development under stress.Both Trp and Ind showed auxin-like effects likely due to their conversion into IAA within the plant. However, further studies carried out under axenic conditions are needed in order to clarify this question.     

番茄植株再生体系的建立

选用杂交番茄早丰(07-13)为材料,以下胚轴、子叶作为外植体,根据控制单一变量与多重比较的原则,设计添加不同种类和不同浓度的植物生长物质的培养基配方,建立了一套简便有效地番茄再生体系。结果表明:愈伤组织的诱导及增殖,以配方MS+6-BA 2.0 mg/L+IAA0.3 mg/L为佳;诱导不定芽的分化,以培养基MS+6-BA 2.0 mg/L+IAA 0.1 mg/L为佳;生根培养基以MS+NAA 0.1 mg/L+IAA 0.1 mg/L为佳,生根率高且根粗壮。     

液相色谱-串联质谱法测定糙皮侧耳中9种植物生长调节剂的残留量

建立了液相色谱-串联质谱仪(Liquid Chromatography-Tandem Mass Spectrometry,LC-MS/MS)同时测定糙皮侧耳(Pleurotus ostreatus)中吲哚乙酸、吲哚丙酸、吲哚丁酸、赤霉素、2,4-二氯苯氧乙酸、6-苄基腺嘌呤、4-氯苯氧乙酸、6-糠氨基嘌呤和芸苔素内酯9种植物生长调节剂残留的定量分析方法,样品经含1%(v/v)乙酸的乙腈溶液提取,于分散固相萃取管(含混匀的150mg MgSO_4和50mg C_(18))净化后用LC-MS/MS检测。结果表明:9种植物生长调节剂的峰面积与其浓度呈良好线性关系,方法最低检出限为0.5~1.0μg/kg;回收率为80.3%~96.5%,相对标准偏差为0.88%~7.31%,该方法准确、可靠,可以同时满足糙皮侧耳中9种植物生长调节剂残留检测的需要。对市场上50份糙皮侧耳的测定结果为:吲哚乙酸检出6次(2.3~96.7μg/kg)、吲哚丙酸2次(21.3、87.8μg/kg)、吲哚丁酸5次(3.4~91.2μg/kg)。     

AgNO_3对草莓试管苗生长及抗氧化酶活性的影响

为了探讨重金属离子对果树的毒性机制,以草莓(Fragaria ananassa Duch．)主栽品种丰香(Toyonoka)和章姬(Akihime)组培苗为外植体,研究了不同浓度AgNO3对草莓生长及抗氧化酶活性的影响。结果表明,1／2MS培养基中有AgNO3存在时,2个草莓品种的根干质量和根长明显增加,根系活力随着AgNO3浓度的增加而增加,1．0 mg／L时达最大值,而根系含水量则随着AgNO3浓度的增加逐渐降低。AgNO3浓度为0．5-1．0 mg／L时,植株株高和干质量逐渐下降,而叶绿素、可溶性蛋白的含量和抗氧化酶的活性则逐渐增加。叶片中MDA的含量、O2-的产生速率、脯氨酸的含量和IAA氧化酶的活性随着AgNO3浓度的增加而增加,而IAA的含量则逐渐降低。     

In vitro adventitious rooting of Cornus florida microshoots

Cornus florida L. (flowering dogwood) has been successfully micropropagated, but low rooting of microshoots makes the system inefficient. This study was conducted to increase rooting efficiency of flowering dogwood microshoots over that previously achieved. Microshoots originating from acclimatized axillary and nodal bud stock cultures were excised and treated with different concentrations and combinations of various auxins including indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA). Effect of microshoot age on rooting efficiency was also examined. Of the auxins tested, maximum rooting was observed with 4.4 μM IBA. The age of microshoot explants had a significant effect on rooting. Five to seven-week-old microshoots treated continuously with 4.9 μM IBA in Woody plant medium (WPM) consistently had the best and most consistent rooting efficiency (about 83%).     

Callus induction and plant regeneration in Cardiospermum halicacabum Linn. an important medicinal plant

Cardiospermum halicacabum Linn. is an important medicinal twining herb belonging to the family sapindaceae. A method for rapid micropropagation of C. helicacabum through plant regeneration from leaf and nodal explant derived calli has been developed. The nodal and leaf segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; 0.5–9 μM) for callus induction. Callus production was highest at 5 μM 2,4-D where 96 and 90% of cultured leaf and nodal cuttings produced callus, respectively. The viable calli were maintained at reduced concentration of 2,4-D (2 μM). These calli were transferred to MS medium supplemented with various concentrations of 6-benzyladenine (BA; 2–10 μM) or kinetin (2–10 μM) alone or in combination with indole 3-acetic acid (IAA; 0.2–1.0 μM) for shoot regeneration. The addition of low concentrations of IAA into BA or kinetin containing medium significantly increased the frequency of shoot regeneration in both nodal cuttings and leaf-derived calli. The highest number of adventitious shoots (28 per callus) formed at 8 μM Kin and 0.5 μM IAA. For rooting of the shoots, half-strength MS medium supplemented with different concentrations of indole 3-acetic acid, indole 3-butyric acid (IBA) and (alpha)-naphthalene acetic acid (NAA) 1–5 μM was tried. The optimal result was observed on half-strength MS medium supplemented with 2.5 μM IBA, on which 91% of the regenerated shoots developed roots with an average of 4.2 roots per shoot within 45 days. The in vitro raised plantlets were acclimatized and transferred to soil with 90% success. This in vitro propagation protocol should be useful for conservation as well as mass propagation of this medicinal plant.     

黄瓜再生体系和除草剂抗性筛选体系的建立

试验以黄瓜自交系C-8子叶节为材料,比较了不同浓度的6-BA和IAA组合对其不定芽诱导的影响,建立了高效快速的再生体系;摸索了不同浓度的草铵膦对产生不定芽的影响,建立了C-8的草铵膦抗性筛选体系,为下一步的遗传转化打下基础。结果表明,最佳的诱芽培养基为MS+0.5mg·L-16-BA+2mg·L-1IAA,出芽率为77.5%,生根培养基为1/2MS。对于C-8黄瓜,草铵膦的最佳遗传转化筛选浓度为0.6mg·L-1。     

Adventitious root formation in Grevillea (Proteaceae), an Australian native species

Grevillea (Proteaceae) is a native Australian plant genus with high commercial value as landscape ornamentals. There has been limited research on the culture and propagation of Australian native species. The effect of indole-3-butyric acid (IBA) on the rooting of G.‘Royal Mantle’ and G.‘Coastal Dawn’ in winter, spring and summer was evaluated at University of Queensland Gatton, Southern Queensland in order to determine the rooting ability of this species in different seasons. Both Grevillea cultivars showed seasonal rooting. The more difficult-to-root G.‘Coastal Dawn’ had a reduced response to IBA application than G. ‘Royal Mantle’. Stem and leaf indole-3-acetic acid (IAA) levels were not different between cultivars, therefore rooting ability between the two cultivars does not appear to be due to the differences in endogenous IAA levels.     

Evaluation,tissue culture propagation,and dissemination of ‘Saba’ and ‘Pelipita’ plantains in Costa Rica

Fruit characteristics of the disease-resistant bluggoe-type (ABB) cooking bananas (Musa × paradisiaca L.) ‘Saba’ and ‘Pelipita’ were compared with those of the susceptible ‘Currare’ (‘Horn’). Yields of ‘Saba’ and ‘Pelipita’ were similar to those of ‘Currare’; however, ‘Saba’ and ‘Pelipita’ yielded fewer hands with a greater number of small fruit when compared with ‘Currare’. Shoot-tip cultures of both clones were readily initiated on a modified Murashige and Skoog (MS) basal medium supplemented with N⁶-benzyladenine (BA) in combination with indole-3-acetic acid (IAA). Propagation cultures were initiated by splitting shoot tips along their longitudinal axis and re-culturing the individual pieces to basal medium supplemented with 5 mg l^?1 BA. Transfer of axillary shoots to hormone-free medium resulted in rapid and extensive root formation. Plantlet survival after transfer to methyl bromide-treated soil exceeded 90%. Establishment in the field was achieved following procedures normally used for vegetative propagation of this crop. Trial plantings of in vitro propagated ‘Saba’ and ‘Pelipita’ were established in various provinces of Costa Rica for grower evaluation and for future comparison of growth and reproductive development with plants of these and other cultivars propagated from corms.     

A simple method for large-scale propagation of Dendrocalamus asper

The influence of position of cuttings, auxin treatments and seasonal variation on adventitious rooting of Dendrocalamus asper, a multipurpose edible bamboo of horticultural value, was examined. Single node proximal (culm) and distal (culm-branch) cuttings were collected during the months of April–June and treated for 24 h with water or 2 mM auxin (indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA)). The treated cuttings were planted horizontally in sand beds and maintained under misting conditions for 2 months. Culm cuttings exhibited better adventitious rooting than culm-branch cuttings and the month of May proved to be the best for rooting of both types of cuttings. Auxin treatments also influenced adventitious rooting and their effectiveness was in the order: NAA>IBA>IAA. The interaction among auxin, position of cuttings and month of collection was also found to be significant on root induction and differentiation. Single node culm cuttings, without auxin treatment, resulted in 98% plantlets in May and 67% plantlets in June, and emerged as a suitable material for large-scale vegetative propagation. Administration of IAA or NAA to abundant culm-branch cuttings of low commercial utility (collected in May) also produces ≥50% plantlets.     

桃Aux/IAA家族基因鉴定及在果实成熟过程中的表达分析

通过生物信息学鉴定桃基因组中生长素/吲哚乙酸(Aux/IAA)基因家族,并对其在溶质型和硬质型桃果实成熟阶段的表达水平进行q RT-PCR检测。结果表明:桃Aux/IAA基因家族包含22个成员,这些基因集中分布在5条染色体上,以第1条染色体上含有最多,为8个。大部分Aux/IAA基因包含高度保守的Ⅰ、Ⅱ、Ⅲ和Ⅳ功能域,聚类分析表明其分为10类。基因结构分析显示这些基因包含2~5个外显子。荧光定量PCR分析表明10个Aux/IAA基因在溶质型桃果实的表达量高于对应时期硬质型桃果实,其中5个基因随着溶质型桃果实的成熟上调表达,特别是ppa010303m、ppa010871m和ppa020369m。试验结果表明桃Aux/IAA家族成员在结构上高度保守,其中多个成员(尤其是ppa010303m、ppa010871m和ppa020369m等)可能参与调控桃果实的成熟。