In vitro morphogenesis of juvenile Annona cherimola Mill,bud explants

A micropropagation procedure for juvenile cherimoya (Annona clierimola Mill.) is described. Axillary shoot proliferation was obtained after culturing nodal sections with lateral buds in basal medium supplemented with either 0.66 μM BA or 1,36μM zeatin. For root induction shoots were pre-incubated for 7 d in basal medium supplemented with 1 gl^?1 activated charcoal, then cultured for 10 d (7 dark/3 light) on medium with 500 μM IBA, 58.4 mM sucrose, and 200 mg I“1 citric acid. Afterwards, shoots were transferred to the same medium without auxin and with the macroelements at half strength. Using this procedure, 95% of shoots rooted. The survival rate at the end of the acclimatization period was 70%.     

阔叶猕猴桃叶片离体器官发生和植株再生(英文)

以阔叶猕猴桃(A ctinidia latifolia M err.)叶片为外植体,通过器官发生途径诱导形成不定芽,探讨了不同激素组合、暗培养时间以及不同浓度的蔗糖对叶片器官发生和植株再生的影响。结果表明,BW +0.1μm ol/L N AA +5μm ol/L Zeatin 附加20g/L 蔗糖,先进行21d 暗培养然后转到光下培养效果最好,6周后不定芽再生频率达91.67%,平均每个外植体再生芽数6.87个。再生芽生根良好,生根率可达100%。首次报道了阔叶猕猴桃的器官发生和植株再生,建立了叶片的高效再生体系,为今后的遗传转化研究奠定了基础。     

余甘子下胚轴离体培养中的器官形成与植株再生(英文)

以余甘子(phyllanthusemblicaL.)下胚轴为外植体,通过器官发生途径诱导形成不定芽,探讨不同激素组合对下胚轴器官发生和植株再生的影响,以期建立有效的再生体系,为以后的遗传转化研究奠定基础。结果表明,在附加0.1μmol/LNAA、5μmol/LBA和30g/L蔗糖MS培养基上培养,5周后不定芽再生频率达87.5%,平均每个外植体再生芽数为9.7个,为最佳组合。将分化的不定芽转移至含有10μmol/LIBA的MS培养基上,5周后生根,发育成健康的植株,生根率在29.2%。     

Direct plant regeneration from mature leaf explants of pistachio, Pistacia vera L.

A protocol was developed for direct shoot and plantlet regeneration from in vitro regenerated leaf explants of male Pistacia vera L. cv. ‘Atl?’. Leaves excised from axenic shoot cultures of pistachio were used to induce organogenesis on a Murashige and Skoog (MS) medium with Gamborg vitamins supplemented with combinations of different concentrations of BAP and IAA. The highest adventitious shoot regeneration in 35% of the explants, with the number of shoots ranging from 2 to 3 per explant, occurred in the explants cultured during the establishment phase in the medium with 1 mg l⁻¹ IAA and 2 mg l⁻¹ BAP. For shoot multiplication, the highest number of new microshoot/explants (5.76) was obtained in a culture medium supplemented with 1 mg l⁻¹ BAP, but it was not significantly different from the number obtained at 2 mg l⁻¹ BAP. A high rooting frequency (84%) for microshoots was recorded on a medium supplemented with 2 mg l⁻¹ IBA. In vitro rooted plantlets were transferred to pots filled with a mixture of soil, sand and peat (1:1:1). They were weaned in a growth room and finally moved to a greenhouse. This protocol could be utilized for in vitro clonal propagation of this economically important plant.     

Clonal propagation of mulberry (Morus indica L. cultivar M-5) through in vitro culture of nodal explants

A high frequency of sprouting (80.0%) and shoot differentiation was observed in the primary cultures of nodal explants of Morus indica L. cultivar M-5 on MS medium supplemented with 2,4-D (0.3 mg/l). In vitro proliferated shoots were multiplied rapidly by culture of shoot tips on MS medium with BAP (0.5 and 1.0 mg/l) which produced the greatest multiple shoot formation. Multiplication was also achieved by culture of shoot tips on MS medium with BAP (4.0 mg/l) and GA₃ (0.05 mg/l) which facilitated the elongation of shoots followed by sprouting of axillary buds of in vitro grown shoots. A high frequency of rooting (86.7%) with development of healthy roots was observed from shoots cultured on medium with 2,4-D (1.0 mg/l). Plants with well developed roots were transferred to soil with a survival frequency of 80%.     

In vitro morphogenesis and growth of Narcissus in response to temperature

In vitro morphogenesis and growth of tissues of the Narcissus cultivar ‘Lord Nelson’ have been assessed in constant and alternating temperatures. Tissue of leaf-base and scape origin, producing leafy shoots and bulblets on a defined sterile medium, were grown at constant and alternating temperatures of 15, 20, 25 or 30°C and 15–25 or 20–25°C, respectively, in a 16-h daily photoperiod. The number of leaves, leaf length, volume of tissues, fresh weight (but not percentage dry matter), and the number of shoots that would ultimately produce bulbs, were maximal at a constant temperature of 25°C as compared with all other temperature-regimes tested.     

Rapid micropropagation of Psoralea corylifolia L. using nodal explants cultured in organic additive-supplemented medium

Summary

The influence of different growth regulators and additives on shoot multiplication from nodal explants of Psoralea corylifolia was investigated. Prolific shoot multiplication was achieved within 4 weeks of culture on Murashige and Skoog (MS) medium supplemented with 5 μM benzyladenine (BA), 5 μM ascorbic acid (AA), 100 mg l^–1 casein hydrolysate (CH) and 5% (v/v) coconut water (CW). Shoots elongated on half-strength MS basal medium devoid of inositol, but containing 5 μM 2-isopentenyladenine (2iP), 10 g l^–1 sucrose and 8 g l^–1 agar. Elongated shoots rooted on half-strength MS basal medium supplemented with 3 μM indole-3-butyric acid (IBA), 10 g l^–1 sucrose and 7 g l^–1 agar within 5 d of culture. The in vitro-raised plants were established successfully in 2:1:1 (v/v/v) garden soil:farmyard soil:sand, and maintained in a growth chamber with 100% survival. Acclimatised plants were transferred to a glasshouse and established successfully in the field. Flowers and fruits appeared after 4 months and resembled those on source plants. This system could be used for rapid commercial propagation of P. corylifolia for conservation strategies and to produce phytomedicines.     

In vitro regeneration from mature leaf explants of Pelargonium rapaceum (L.) L’Hérit

Methods to regenerate whole plants from mature leaf explants of Pelargonium rapaceum (L.) L’Hérit were established. To optimize shoot induction, leaf explants were cultured on media containing different types and combinations of plant growth regulators. Growth was initiated within 17–24 days culture, and included callus formation, and root or shoot organogenesis ranging from 20 to 100% regeneration. Shoots were induced only when explants were cultivated on MS medium containing a combination of NAA and kinetin, NAA and BAP, IAA and Kinetin, or IAA and BAP. On media containing NAA and BAP, dark incubation was critical for efficient direct shoot regeneration from explants. Direct shoot formation and the highest number of shoots per explant (17.6) were obtained from leaf explants cultured in the dark for 30 days on MS medium containing 0.1 mg l⁻¹ NAA and 0.1 mg l⁻¹ BAP. Shoots cultured on MS medium containing 0.1 mg l⁻¹ NAA formed tuberous roots with microtubers within 42 days. Healthy regenerated plants were acclimated and transferred to a greenhouse.     

阿月浑子外植体褐变抑制方法

外植体褐变是阿月浑子组培的主要障碍之一。对阿月浑子外植体母树年龄、遮光、外植体枝条部位与采集季节、组培条件、添加剂以及多酚氧化酶活性等与外植体褐变的关系进行了研究,结果表明,阿月浑子外植体的褐变率夏季最高、秋季其次、春季最低,枝条中部最低,而枝条上端最高,1年生与7年生树褐变率差异不大,与母树是否遮光关系不明显,在培养基中添加抑制剂PVP能减轻褐变,而添加维生素C、Na2S2O3与活性碳的效果不显著,低光和低温对控制外植体褐变有显著效果,但从低温下转入常温后褐变率高于一直在常温下培养的,阿月浑子褐变率与多酚氧化酶活性关系不显著,可能主要决定于外植体多酚含量,但需要进一步实验数据支持。     

Effect of clinorotation on in vitro cultured explants of Mentha piperita L

An in vitro culture system was used to study the influence of gravity on axillary shoot formation and adventitious root regeneration in Mentha piperita L. The direction of the gravity vector was altered by displacing stem node explants in different orientations. Also, microgravity conditions were simulated by rotating the explants on a horizontal clinostat so that the main axis of nodes was either parallel (Cpa) or perpendicular to the clinostat axis (Ccp and Ccf, centripetally and centrifugally oriented, respectively). Mint nodes were cultured on solidified Linsmaier and Skoog's medium [Physiol. Plant. 18 (1965) 100] adding a filter-sterilized aqueous solution of 2 mg/l benzyladenine (BA) in half of the cultures. The proliferation of axillary shoots as well as adventitious root formation were not affected by altering upright explant orientation. On the contrary clinorotation was able to modify plantlet development. In absence of BA, leaf width was hindered by Cpa treatment and penultimate internode length was enhanced by Ccp. Furthermore, a negative effect of Cpa treatment was observed in root length parameter, while Ccp increased the root number both in absence and in presence of BA. An effect strictly connected to clinorotation in presence of BA was the occurrence of hyperhydricity. Moreover, explants under clinorotation treatments switched their gravitropic response modifying shoot curvature.     

硝酸银和硫代硫酸银对籽瓜强雌系的诱雄效果

瓜类强雌系可用于简化人工杂交制种,通过选取硝酸银和硫代硫酸银作为诱雄剂,对籽瓜强雌系幼苗进行诱雄处理,为籽瓜强雌系扩繁保存提供理论依据。以籽瓜强雌系‘DK1’为试材,采用不同浓度的硝酸银(100、300、500、700 mg·L-1)和硫代硫酸银(300、600、900、1 200 mg·L-1)溶液在幼苗第1～3片真叶平展期分别进行不同喷施次数(1次、2次、3次)处理,统计比较了2种诱雄剂不同浓度及次数处理后植株主蔓第5～20节位内两性花、雄花、雌花和败育花的数量。结果表明,300 mg·L-1硝酸银和900 mg·L-1硫代硫酸银喷施3次诱导两性花的效果最佳,其中硫代硫酸银溶液的诱导两性花数量优于硝酸银溶液。生产上可参考此诱雄方法扩繁籽瓜强雌系。     

辣椒单倍体离体诱导及育种应用

 概括介绍了辣椒单倍体诱导的重要进展,对影响花药和小孢子培养的材料、预处理、培养基种类、离体雄核发育的细胞学标记及过程,以及单倍体诱导技术在辣椒杂交育种中的应用进行了综述。     

In vitro micrografting and its applications to fruit science

Improvements have been brought to in vitro micrografting. The breaking of dormancy by dipping the seeds of the rootstock in concentrated solutions of benzylaminopurine avoids having recourse to long stratification in the cold (60–120 days according to species). The initial pre-treatment of apices with cytokinin allows their development and facilitates later grafting. Moreover, adding DIECA to the cut sections avoids oxidation and, together with the insertion of a piece of agar between apex and rootstock, increases the success of micrografting. Finally, a special technique of in vivo micrografting of pretreated apices on young plants has been developed.Micrografting is commonly used to obtain peach trees free of Sharka and NRS viruses, and to study incompatibility mechanisms in heterografts, leading to an early diagnosis of incompatibilities.     

Hormone effects on in vitro proliferation and rooting of Grevillea explants

Explants of Grevillea rosmarinifolia A. Cunn. and Grevillea×semperflorens F.E. Brigs ex Mullig., obtained from vegetative stem segments, were grown on a modified MS medium, which was supplemented with different contents of cytokinins (BA or K) to evaluate optimal conditions for bud proliferation. Auxins (NAA or IBA) were tested to evaluate rooting.

The multiplication rate was often higher in G. rosmarinifolia than in G.×semperflorens. Explant activity varied significantly depending on the cytokinin type and its concentration. The addition of cytokinins to the culture medium often promoted an increase in the number of shoots produced by each propagule, but at the highest BA concentration, shoot differentiation was inhibited in Grevillea×semperflorens. Only the higher concentrations studied (5–10 mg/l) produced better results with K, whereas BA was more effective at 1–2 mg/l. Rooting efficiency of the young shoots was lower in G. rosmarinifolia than in G.×semperflorens. IBA promoted root development in both species and was more effective at the highest concentration (1 mg/l). NAA gave better results at low concentrations (0.2 mg/l) in G. rosmarinifolia, while it had slight or negative effect on rooting of G.×semperflorens.     

硝酸银和赤霉素对西瓜雌性系诱雄效果的影响

为了探索硝酸银和赤霉素对西瓜雌性系诱雄效果及生长发育的影响,以雌雄异花同株材料‘XHB’和‘TS409’及其血缘相同的两个雌性系突变体‘XHBGM’和‘TGS409’为试材,于春、秋两季,在西瓜苗期喷施不同质量浓度的硝酸银(100、300、500 mg·L~(-1))和赤霉素(500、1 000、1 500 mg·L~(-1)),调查了处理后植株高度、茎粗度等生长指标及各单株前30个节位内雄花、雌花、两性花的数量。结果表明,赤霉素可促使雌雄异花同株材料产生更多的雄花,但在雌性系材料上无明显的诱雄效果,植株表现徒长现象,植株高度、节间长度明显增加,茎粗度、叶片叶绿素含量显著降低。硝酸银在雌性系和雌雄异花同株材料上均可诱导产生两性花,秋季效果更为明显,且质量浓度越高,诱雄效果越好,但高质量浓度的硝酸银对植株生长产生抑制作用,甚至导致植物死亡;300 mg·L~(-1)是诱导西瓜雌性系有效产生两性花的最佳质量浓度,对植株伤害较小,秋季诱雄效果更佳,生产上可采用此方法来繁殖西瓜雌性系。     

Rootstock influences the response of pistachio (Pistacia vera L. cv. Kerman) to water stress and rehydration

Pistachio cultivation requires the use of rootstock because grafting is the only form of vegetative propagation. The main commercial rootstocks are Pistacia integerrima L., Pistacia atlantica Desf., Pistacia terebinthus L. and Pistacia vera L. Pistachio is considered to be a drought and saline-resistant crop; however, there is little information describing varietal responses of rootstocks to water stress. Some studies have suggested that P. terebinthus L. is the most drought and cold resistant rootstock. The effect of the rootstock on the water relations of the grafted plant is crucial for improving crop performance under water stress conditions and for developing the best irrigation strategy. This work studied the physiological response to water stress of pistachio plants (P. vera L. cv. Kerman) grafted onto three different rootstocks P. terebinthus L., P. atlantica Desf. and a hybrid from crossbreeding P. atlantica Desf. × P. vera L. Plant physiological responses were evaluated during a cycle of drought and subsequent recovery in potted plants. Parameters measured were soil moisture, trunk diameter, leaf area, leaf number, leaf and stem dry weight, stem water potential, leaf stomatal conductance. The results showed different responses of cv. Kerman depending on the rootstock onto which it had been grafted. The hybrid rootstock was associated with a higher degree of stomatal control and reduced leaf senescence compared to P. atlantica and P. terebinthus, despite being associated with the most vigorous shoot growth. P. terebinthus enabled very effective stomatal control but was also associated with the most rapid leaf senescence. P. atlantica was associated with less vigorous shoot growth and similar levels of water stress as occurred with the others rootstocks under conditions of high evaporative demand, which was associated with lower stomatal control. The selection of the most effective rootstock choice for different environmental conditions is discussed.     

梨外植体组培褐变的影响因子及预防措施

以鸭梨、黄金梨、阿巴特梨和杜梨为试材,对梨组培过程中影响外植体褐化的因素及防褐措施进行了研究。结果表明,品种、采样时期以及外植体内酚类物质含量等因素都显著地影响材料褐化率。抗褐剂试验表明,培养基中添加0.2g/L的聚乙烯吡咯烷酮(PVP)或100mg/L抗坏血酸(Vc),或将外植体在200mg/L抗坏血酸水溶液中浸泡30min后接入添加2g/L活性炭的培养基,均能显著抑制鸭梨褐化的发生。低温试验表明,鸭梨外植体经4℃低温处理6h后接种,或接入初期在4℃低温中培养12～24h,褐化程度明显减轻。     

In vitro micropropagation of Ephedra

The in vitro micropropagation of eleven species of Ephedra was investigated. Shoot nodal explants of E. fragilis were cultured on Murashige and Skoog medium supplemented with 3% sucrose, 0.05 jiM 3-indolebutryic acid and 0.0-0.5 |iM kinetin, zeatin or 6-ben- zylaminopurine. In general, the average number of shoots produced per explant increased and the average shoot length decreased with increasing cytokinin concentration. Substitution of 3-indolebutryic acid with 2,4-dichlorophenoxyacetic acid caused callusing and distorted shoot growth. Shoot cultures of ten other species were grown on 0.05 |iM 3-indolebutyric acid with 0.05 kinetin. Indole-3-acetic acid gave healthy rooting. E. equisitina, E. gerardiana, E. minima and E. saxatilis were successfully micropropagated using a single-stage protocol in which shoots were grown and rooted on Sorbarods using half strength Murashige and Skoog medium supplemented with 1% sucrose and 5.0 (iM indole-3-acetic acid. Healthy plantlets were weaned in John Innes No. 1: Perlite (1:1) following treatment with Captan (1.9 g I'1).     

In vitro propagation of spathiphyllum

An in vitro method for propagation of Spathiphyllum cultivar ‘Clevelandii’ is described as an alternative vegetative-propagation method.Leaves, inflorescences, peduncles, buds and stem pieces were tried as explants. Initiation and development of shoots did not occur when leaves and peduncles were used. In a few cases, it was possible to induce shoots from pieces of inflorescence explants. Buds and especially stem pieces were very suitable as explants, with shoots developing on the basal medium both with or without a cytokinin.Further multiplication of the shoots was optimal on the basal medium supplemented with 2 mg/l PBA. On this medium, the basal part of the old shoots became swollen and callus-like, and new shoots emerged from the callused area.Shoots developed roots very easily on the basal medium without hormone additives.     

In vitro propagation of watermelon

A 3-stage, in vitro propagation system for diploid watermelon (Citrullus lanatus (Thumb.) Matsum. and Nakai) has been developed which is also applicable to the economic production of triploid (seedless) watermelon transplants.Stage I involves the stimulation of axillary-bud development from excised seedling shoot tips (1–3 mm) by a high cytokinin (K)/low auxin (IAA) ratio (4.6 μmol/l K 0.28 μmol/l IAA) on a modified Linsmaier and Skoog medium. An average of 4.5 axillary shoots of sufficient size to subculture (> 15 mm) were obtained from each explant in 5 weeks. These shoots were induced to root during 3 weeks subculture on Stage II medium containing 11.5 μmol/l IAA. Well-rooted plantlets were successfully transplanted to a 1 : 1 peat : sand (v/v) substrate and gradually “hardened-off” to greenhouse conditions (Stage III) for 3 weeks, after which they could be transferred to field conditions.Alternatively, axillary shoots from Stage I could be returned to fresh media with 9.3 μmol/l K and 0.28 μmol/l IAA, where an average of 10.3 axillary shoots could be obtained after 5 weeks.Cost estimates for producing 10 000 finished transplants per week project an approximate cost of 16 dollar cents per transplant.