Metabolic changes during rooting in stem cuttings of five mangrove species

Vegetative propagation through rooting in stem cuttings in five tree mangroves namely Bruguiera parviflora, Cynometra iripa, Excoecaria agallocha, Heritiera fomes, and Thespesia populnea using IAA, IBA and NAA was reported. Spectacular increase in the root number was noted in the cuttings of H. fomes and C. iripa treated together with IBA (5000 ppm) and NAA (2500 ppm). The highest number of roots was obtained with IBA (2500 ppm) and NAA (500 ppm) in E. agallocha. B. parviflora and T. populnea responded better to IAA and IBA treatment. The species specific variation in the rooting response to exogenous application of auxins was reflected in the metabolic changes during initiation and development of roots in cuttings. Biochemical analysis showed increase of reducing sugar in the above-girdled tissues at initiation as well as subsequent development of roots which was further enhanced by the use of auxins. Decreases in the total sugar, total carbohydrate and polyphenols and increase in total nitrogen were recorded in the girdled tissues and the high C/N ratio at the initial stage helped in initiation of roots in all the species. Interaction of IBA and NAA promoted starch hydrolysis better than IAA and IBA during root development and subsequently reduced the C/N ratio and increased the protein-nitrogen activity during root development which suggest the auxin influenced mobilization of nitrogen to the rooting zone.Abbreviations IAA Indole-3-acetic acid - IBA Indole-butyric acid - NAA A-naphthalene acetic acid     

Auxin Synergists in Rooting of Cuttings

Leafy cuttings of Eranthemum tricolor were treated with tannic acid, gallic acid, p-hydroxybenzoic acid and salicylic acid at the concentrations of 1000, 100, 10 and 1 nig/1 for 24 hours, whereafter they were dipped quickly in a 1000 mg/l solution of IAA, IBA and NAA for ten seconds. None of the phenolics showed any root promoting effect when used singly. In combination with NAA and IBA tannic acid promoted rooting, however, with IAA there was no effect to be seen. Gallic acid also markedly increased the number of roots of cuttings treated with NAA and IBA. Even in this case there was no effect with IAA. Synergism was also recorded between p-hydroxybenzoic acid and IAA or NAA but not with IBA. Salicylic acid greatly promoted rooting in combination with both IAA, IBA and NAA.     

High Frequency Clonal Propagation of Cymbopogon martinii var motia (palmarosa) Through Rhizome Culture and True to Type Assessment using ISSR Marker

A rapid and high frequency plant propagation system has been established in Cymbopogon martinii var motia using rhizome culture. Different concentrations of auxins, such as 5.36 μM Napthalene acetic acid (NAA) and 5.71μM Indole 3acetic acid (IAA) satisfactorily induced shoot buds when applied individually or in combination with 4.40μM N⁶-Benzyladenine (BA) on MS medium. Multiple shoot proliferation was noted when induced microshoots were cultured on MS medium supplemented with BA either alone or with NAA. Shoot bud induction and multiple shoot formation were enhanced significantly with the application of growth additives like coconut water (CW) and biotin. Out of the two auxins (IAA and IBA) tested, rooting percentage and number of roots/shoot was significantly higher on 1/2 MS medium supplemented with 4.90 μM Indole 3- butyric acid (IBA). Twelve reproducible ISSR primers efficiently screened 16 randomly chosen regenerants of C. martinii with similar monomorphic banding profiles exhibiting genetic stability of the regenerants.     

Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of <Emphasis Type="Italic">Encyclia mariae</Emphasis> (Orchidaceae), a threatened Mexican orchid

Two procedures for the in vitro propagation of Encyclia mariae, a threatened Mexican orchid, were developed. In the first procedure, leaves from in vitro germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with the range of 2.21–4.4 μM 6-benzylaminopurine (BA) in combination with 2.69–10.74 μM naphthalene acetic (NAA), 2.07–8.29 μM indole-3-butyric (IBA), or 2.85–11.42 μM indole-3-acetic acid (IAA) to determine the best medium for the induction of shooting. Maximum direct shoot formation from leaves was observed on MS containing 22.21 μM BA and 10.74 μM NAA (25 shoots/explant). The second procedure began with the culture of protocorms on media containing NAA, IBA, or IAA, which induced callus formation with high regenerative potential in the form of protocorm-like-bodies (PLBs) that eventually differentiated into shoots. The optimal response was attained when these structures were cultured on medium with 4.14 μM IBA (30 shoots/PLB). To promote the elongation of shoots derived from PLBs, the material was subcultured onto MS medium containing 22.21 μM BA and 5.37 μM NAA. Through the exploration of the effects of auxins and matrix on the rooting of shoots, it was determined that the optimal rooting occurred on media supplemented either with 5.71 μM IAA or 4.14 μM IBA either on agar-gelled medium or in liquid media with coir as the matrix. Rooting was found to be 20% higher in liquid media than in agar-gelled medium.     

Auxin stability and accumulation during in vitro shoot morphogenesis influences subsequent root induction and development in <Emphasis Type="Italic">Eucalyptus grandis</Emphasis>

Recent results showed that after 16 months in the field, micropropagated eucalyptus plants have an inferior root system to cuttings. Such differences may be due to the plant growth regulators supplied during the culture stages of standard protocols, which are targeted at optimising plantlet yields and not root quality. This study investigated such a proposal, focusing on auxins in an easy-to-root clone. Initial results showed that the auxin provided in the standard protocol (NAA for multiplication and IBA for elongation) enabled 100% rooting in auxin-free medium, where rooting was faster than on IBA-rooting media. When auxin supply was omitted from multiplication and restricted to NAA or IAA during elongation, rooting in an auxin-free medium was reduced to 68 and 31%, respectively, reflecting the stabilities of these auxins in plant tissues. Additionally, 15% of shoots from the NAA-medium and 65% from the IAA-medium produced roots with altered graviperception. GC–MS analysis of these shoots revealed a relationship between free IAA-availability and altered graviperception. This was further tested by adding the IAA-specific transport inhibitor 2,3,5-triiodobenzoic acid to rooting media with IBA, IAA or NAA, which resulted in 100, 70.9 and 20.6% rooting, respectively. At least 40% of the sampled root tips had atypical starch grain deposition and abnormal graviperception. It is proposed that, at least in this clone, while IBA and NAA can be used for in vitro root induction, IAA is necessary for development of graviresponse.     

植物生长调节剂对几种园林植物插条生根的影响

 我们用NAA、IBA、IAA等植物生长调节剂促进多种园林植物插条发根的试验。结果表明：菊花、大丽花插枝,经0.1mg/L和0.2 mg/L NAA处理,具有促进发根的明显效果;0.1mg／L和0.2 mg/L的NAA和IBA处理较难发根的印度黄檀插枝,其发根率则有显著提高;月季、大叶黄杨、水杉、池杉、海南石梓等植物插枝,虽都具有较高的发根率,但经不同浓度NAA和IBA浸渍处理后,其发根率仍有不同程度的提高。     

Effect of Auxins on Adventitious Root Development from Single Node Cuttings of Camellia sinensis (L.) Kuntze and Associated Biochemical Changes

An attempt was made to induce rooting from single node cuttings of Camellia sinensis var. TV-20 under controlled conditions and study its biochemical changes during rooting. The nodal cuttings were pretreated with different concentrations of IAA, NAA and IBA and kept in a growth chamber (25 ±2 °C, 16 h photoperiod (55 μ mol m⁻² s⁻¹) with cool, white fluorescent lamps and 65% relative humidity) for 12 h. Among the three auxins used for pretreatment, IBA showed more positive response on rooting as compared to IAA and NAA within 2 weeks of transfer to potting medium. Among four concentrations of IBA tested, 75 ppm gave maximum percentage of rooting, number of roots and root length. Therefore, IBA was used further in experiments for biochemical investigation. The adventitious rooting was obtained in three distinct phases i.e. induction (0–12 days), initiation (12–14 days) and expression (14–18 days). IAA-oxidase activity of IBA-treated cuttings increased slightly as compared to control. The activity was found to decrease during induction and initiation phases and increase during expression phase. The peroxidase activity in IBA-treated cuttings increased up to initiation phase and declined at the expression phase. Polyphenoloxidase activity increased both in IBA-treated and control cuttings during induction and initiation phase but declined slowly during expression phase. Total phenolic content was higher in IBA-treated cuttings, particularly in initiation and expression phases and it also correlated with peroxidase activity. Phenolics might be playing key role for induction of adventitious rooting, and phenolic compounds can be used as rooting enhancer in tea plant.     

Shoot organogenesis and plant regeneration in <Emphasis Type="Italic">Metabriggsia ovalifolia</Emphasis>

An efficient propagation and regeneration system via direct shoot organogenesis for an endangered species, Metabriggsia ovalifolia, was established. High activity cytokinins [6-benzyladeneine (BA) and thidiazuron (TDZ)] and low activity auxins [α-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA)] could directly induce adventitious shoots from leaf or petiole explants within 5 weeks. Cytokinins (TDZ or BA) combined with auxin (NAA) in the induction media induced more adventitious shoots than when auxins or cytokinins were used alone. Adventitious shoots could be induced and also mass-propagated on media containing 2.5–5.0 μM TDZ (or BA) and 0.25–0.5 μM NAA. Adventitious roots differentiated at the proximal end of shoots on rooting media containing half-strength MS salts and 0.5 μM IBA, 0.5 μM NAA, 0.1% activated charcoal or no plant growth regulators. Over 90% of plantlets survived following acclimatization and transfer to a potting mixture (1:1, sand:vermiculite) in basins.     

Integration of trichoderma pseudokoningii with agrochemicals for disease management and plant development in mulberry

Vegetative propagation of mulberry (Morus spp.) is greatly afflicted by soilborne diseases. Stem canker (Botryodiplodia theobromae) and cutting rot (Fusarium solani) are the major diseases which cause high rate of mortality of stem‐cuttings (vegetative clones), thus affecting the initial establishment of mulberry. Apart from diseases, the poor rooting ability of stem‐cuttings of many promising mulberry varieties also leads to unproductive propagation. An effective biocontrol agent (Trichoderma pseudokoningii), chemical fungicide (Dithane M‐45) and phytohormones (Indole‐3‐acetic acid, IAA and 1‐Naphthalene acetic acid, NAA) were evaluated individually and in combination for their effects on disease control and plant development. In plates, T. pseudokoningii was found to be compatible with Dithane M‐45 at 0.1%, and IAA and NAA at 20 ppm concentrations. Under field conditions, the integration of soil application of T. pseudokoningii (in the form of its formulated product, ‘NURSERY‐GUARD') and dipping of stem‐cuttings in 0.1% solution of Dithane M‐45, or 20 ppm solution of IAA showed better results than their individual application. NURSERY‐GUARD with Dithane M‐45 (0.1%) or IAA (20 ppm) provided 47.6 to 52.4% disease control to stem‐cuttings which resulted in significant increase (over check) in sprouting of stem‐cuttings (53.1–63.3%) and plant survival (52.6–57.8%).     

An efficient in vitro propagation protocol for <Emphasis Type="Italic">Pinguicula lusitanica</Emphasis>, a rare insectivorous plant

In this study, an efficient protocol was developed for in vitro propagation of Pinguicula lusitanica L., a rare insectivorous plant with pharmacological value and limited reproductive capacity. The effects of two concentrations (0.1 and 0.5 mg l⁻¹) of a range of plant growth regulators, including cytokinins (BA, KIN, and ZEA) and auxins (IAA, IBA, and NAA), and three concentrations of MS medium macronutrients (total, 1/2 and 1/4MS) on proliferation and rooting, were investigated. P. lusitanica shoots showed abundant proliferation and rooting capacity, both of which were significantly influenced by MS medium strength. The use of 1/2MS supplemented with 0.5 mg l⁻¹ BA or KIN ensure a 29-fold rate of proliferation. Best rooting frequency and higher root number and length were attained in 1/4MS medium containing 0.2 mg l⁻¹ IAA. Sixty percent of the plantlets were successfully acclimatized to ex vitro conditions, exhibiting normal development. These plantlets can be used to replenish declining populations in the wild and/or for the extraction of bioactive compounds, reducing pressure on wild stocks.     

IBA和NAA对山杜英组培苗生根过程中内源IAA、ABA含量变化的影响

本文研究山杜英组培苗生根过程中内源IAA、ABA含量变化规律。结果表明,培养基添加IBA和NAA后,在生根过程中内源IAA、ABA含量变化类似,根点出现前内源IAA、ABA含量一直上升,根点出现后含量开始下降,产生愈伤组织时两种处理的IAA/ABA分别是2.526和3.226。在不添加外源生长素情况下,内源IAA含量一直维持在较低水平,而内源ABA含量一直呈现上升趋势,IAA/ABA始终都在1.211以下。     

Shoot regeneration from cotyledonary leaf explants of <Emphasis Type="Italic">Jatropha curcas</Emphasis>: a biodiesel plant

A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP, and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 μM BAP and 8.5 μM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l⁻¹ activated charcoal. Elongated shoot treated with 15 μM IBA, 5.7 μM IAA, and 11 μM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification.     

渐狭叶烟草组培苗生根条件优化及在基因转化中的应用

针对渐狭叶烟草(Nicotiana attenuata)组培苗生根困难的问题，本研究测试了不同浓度萘乙酸(NAA)、吲哚丁酸(IBA)、吲哚乙酸(IAA)、嘧啶醇、矮壮素、生根粉及活性炭等外源添加物对组培苗生根的影响。结果表明，对照处理的组培苗在90 d内基本不生根；不同浓度矮壮素处理也不能诱导组培苗生根；而合适浓度的NAA、IBA或IAA处理均能较好地诱导组培苗生根，其中0.8 mg·L^-1 IBA不仅具有较好的诱导生根效果，生根率达72.73%，且根及地上部分均生长良好。进一步测试发现0.8 mg·L^-1 IBA处理对转基因组培苗具有良好的生根效果，20 d后生根率达60%以上，而对照组未生根；经伤口诱导和0.8 mg·L^-1 IBA处理，40 d内总生根率可达90%以上。该研究为高效获得渐狭叶烟草转基因生根苗提供了解决方案，也为渐狭叶烟草作为烟草属模式植物的研究提供了便利。     

In vitro regeneration of the tropical multipurpose leguminous tree Sesbania grandiflora from cotyledon explants

Summary A system using cotyledon pieces as explants and a BAP/NAA containing medium was developed for in vitro mass propagation of Sesbania grandiflora, a tropical nitrogen-fixing leguminous tree. The age and the lighting conditions of seedlings providing the explants were shown to be critical factors for both bud induction and bud elongation. Optimal choice for an efficient and reproducible bud induction process consisted of dark-grown seedlings, 24/36 h-old-post-imbibition, that yielded up to 96% of explants producing more than 30 buds each, after one week in culture. Bud development occurred throughout a direct organogenesis pathway, from the proximal and adaxial cut surface of the explants as proved by histological studies. Additional sites of regeneration were also obtained after wounding on the epidermal surface of explants, suggesting a large distribution of regenerative cells all along the explants. Bud elongation, i.e. stem differentiation and leaf growth, was improved by bud isolation from cotyledon explants and their further subculture in liquid bud elongation media for one week. Rooting was obtained on an auxin medium after 3 weeks and plants were established in soil with 92% success.Abbreviations BAP 6-BenzylAminoPurine - NAA a-Naphthalene Acetic Acid - IBA Indole-3-Butyric Acid - IAA Indole-3-Acetic Acid - GA3 Gibberellic Acid - MS Murashige and Skoog (1962) medium     

外源激素处理对三峡消落带落羽杉扦插生根的影响

为优选落羽杉大龄母树插穗的扦插繁殖技术,以期培育大量的落羽杉良种壮苗满足三峡消落带植被修复的需要,采用正交试验设计,探究不同外源激素、浓度以及处理时间对三峡消落带落羽杉大龄母树插穗扦插生根的影响,运用隶属函数法对各处理生根情况进行综合评价。结果表明：（1）落羽杉插穗皮部和愈伤组织处均有不定根伸出,两处不定根数量分别占不定根总数的63.9%和36.1%;（2）4种外源激素中,吲哚丁酸（IBA）+萘乙酸（NAA）（等质量比）和IBA处理的插穗生根效果最好,NAA处理次之,生根粉（ABT）处理效果最差;（3）5种浓度（50 mg/L、100 mg/L、150 mg/L、200 mg/L和250 mg/L）之间的处理效果无显著差异;（4）4种处理时间（2 h、4 h、6 h和8 h）中,4 h处理的插穗生根效果最佳;（5）26个处理组合中,（IBA+NAA）×150 mg/L×4 h处理组合和NAA×250 mg/L×4 h处理组合的平均隶属函数值最高,分别为0.83和0.82,清水对照的平均隶属函数值最低,为0.05。研究初步验证了皮部生根是落羽杉大龄母树插穗的主要生根方式,筛选出三峡消落带原位适生落羽杉大龄母树插穗扦插的两种较佳处理组合为（IBA+NAA）×150 mg/L×4 h和NAA×250 mg/L×4 h。     

Effect of auxins and plant oligosaccharides on root formation and elongation growth of mung bean hypocotyls

The interaction of auxins – IAA, IBA or NAA – with galactoglucomannan oligosaccharides (GGMOs) on adventitious root formation and elongation growth of mung bean hypocotyl cuttings was studied. GGMOs induced adventitious roots in the absence of auxins; however, their effect was lower compared with IBA or NAA. On the other hand, in the presence of auxins, GGMOs inhibited adventitious root induction. Their effect depended on the concentration of oligosaccharides and the type of auxin used. The highest inhibition effect of GGMOs at a concentration of 10⁻⁸ M in the presence of IBA and NAA was observed. In the presence of IAA their inhibition was non-significant in regard to the concentration. The interaction of auxins with GGMOs resulted in the formation of adventitious roots on a shorter part of hypocotyls compared with the effect of auxins alone. However, roots were induced more extensively along the hypocotyls treated with GGMOs compared with the control. GGMOs inhibited the length of induced adventitious roots in the presence of IAA, while in combination with IBA or NAA they were ineffective. The elongation of hypocotyls induced by IAA or IBA was inhibited by GGMOs, too. However, in the presence of NAA or by endogenous growth they were without any significant effect on elongation growth. These findings suggest that GGMOs in certain concentrations might inhibit rooting and the elongation process dependant on auxin used.     

Improving propagation methods of Ricinodendron heudelotti Baill. from cuttings

Ricinodendron heudelotii is a valuable multipurpose tree from Africa that is lacking an efficient and inexpensive vegetative propagation method. To improve multiplication, a series of nursery experiments were conducted to assess the effect of propagation media, plant growth regulators (NAA and IBA), accession source harvest timing, cutting type, and pre-treatment with honey. Maximum survival and rooting percentages of 90.2 and 93.7% respectively were achieved in all experiments. The maximum number of leaves and roots were respectively, 7.4 and 8.7; maximum root length was 15.6 cm. Fine sand was superior to sawdust or fine sand:sawdust mixture. IBA, at 100 μg/l was more effective than NAA for rooting and growth. Principal component analysis showed that, independent of the accession source, cuttings harvested from April to May were more suitable for propagation, giving the highest rates of rooting and growth. In contrast, variable results were achieved with apical cuttings (highest rooting), basal single-node leafy cuttings (highest number of leaves and length of root) and basal two-node leafy cuttings (highest number of roots). Cuttings were more successful when they were soaked in honey for 60 min. After acclimatization, maximum survival rate of plants was 67.6%. This improved protocol can be incorporated into agro forestry system in which propagation of R. heudelotii is a component.     

Root Development Enhanced by Using Indole-3-butyric Acid and Naphthalene Acetic Acid and Associated Biochemical Changes of In Vitro Azalea Microshoots

Based on the importance of producing in vitro adventitious roots, this study was carried out to investigate the effects of indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA) at a concentration of 2 mg L^?1 on the formation of adventitious roots of azalea and their impact on biochemical changes and endogenous hormones. The rooting percentage, root number, and root length were increased in the microshoots of both studied cultivars (‘Mingchao’ and ‘Zihudie’) when the growth medium was supplemented with IBA. Additionally, peroxidase, indole acetic acid oxidase, hydrogen peroxide, and soluble protein contents were improved in both cultivars by auxin treatments especially during the first 7 days of the rooting period. However, application of IBA and NAA increased catalase and polyphenol oxidase in both cultivars during the first 14 and 28 days of culture. The increase in endogenous indole acetic acid (IAA) levels was accompanied by low activity of IAAO during most periods of root induction of microshoots in all treatments. Endogenous gibberellic acid levels were increased after 7 days of culture and then increased again after 28 days of culture. In contrast, the levels of endogenous zeatin riboside and isopentenyl adenosine were decreased with auxin treatments in the first period of the rooting process and then increased after 21 and 28 days of culture. The present study demonstrated that IBA at a concentration of 2 mg L^?1 has a strong effect on azalea rooting. Moreover, the efficiency of IBA and NAA effects on biochemical changes during adventitious root induction was investigated, which may provide new horizons of in vitro rooting production and provide valuable information for the micropropagation of Rhododendron plants.     

Changes in peroxidase activity,auxin level and ethylene production during root formation by poplar shoots raised in vitro

Shoots of poplar (Populus tremula × P. tremuloïdes) were multiplied in vitro and rooted on a rooting medium in the presence of NAA. No rooting occurred in the absence of exogenous auxin. A peak of soluble peroxidase activity, which corresponded to a decrease in the free IAA level in the shoots, preceded rooting These events were considered as corresponding to the initiative phase of rooting. They are preceded by a peak in free IAA activity which might initiate the inductive phase of the rooting process. A burst of ethylene production was measured in both rooting and non-rooting shoots, but the ethylene peak from rooting shoots appeared earlier and was higher. The use of ACC indicated that the exogenous auxin might have enhanced ACC-synthetase activity.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - NAA naphthaleneacetic acid - IAA indole-3-acetic acid - 2-iP 2-isopentenyladenine - IAAsp indole-3-acetylaspartic acid - IBA indole-3-butyric acid - GC gas-chromatography     

Chitosan and silver nanoparticles are attractive auxin carriers: A comparative study on the adventitious rooting of microcuttings in apple rootstocks

Nanocarriers for encapsulation and sustained release of agrochemicals such as auxins have emerged as an attractive strategy to provide enhanced bioavailability and efficacy for improved crop yields and nutrition quality. Here, a comparative study was conducted on the effectiveness of chitosan-as a biopolymeric nanocarrier- and silver-as a metallic nanocarrier- on in vitro adventitious rooting potential of microcuttings in apple rootstocks, for the first time. Auxins indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) loaded silver (nAg) or chitosan nanoparticles (nChi) were synthesized. Scanning electron microscopy and transmission electron microscopy studies showed the spherical shape of the nanoparticles. The average particle size of IAA-nChi was 167.5 ± 0.1 nm while that of IBA-nChi was 123.2 ± 2.6 nm. The hydrodynamic diameter of the nAg-IAA and nAg-IBA particles were measured as 93.66 ± 5 nm and 71.41 ± 3 nm, respectively. Fourier transform infrared spectroscopy analyses confirmed the encapsulation of IAA or IBA in the chitosan nanoparticles. Meanwhile, the characteristic peaks of IAA or IBA were detected on silver nanoparticles. In-vitro adventitious rooting of microcuttings of Malling Merton 106 (MM 106) was significantly higher both in chitosan and silver nanoparticles loaded with IAA or IBA (91.7%–62.5%) compared to free IAA or IBA applications (50.0%–33.3%), except for 2.0 mg L^–1 IBA (66.7%). However, the application of 2 mg L^–1 IBA and IBA-nChi at all concentrations caused an undesirable large callus development.