Shoot Induction Rate Research Articles

In vitromass propagation and acclimatization ofHaworthia truncata

The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA 1 ~ 2 mgL-1 treatment, and NAA 1 mgL-1 + TDZ 2 mgL-1 treatment. The flower stalk showed 75% callus formation rate, at NAA 2 mgL-1 + TDZ 2 mgL-1 treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA 0.1 mgL-1 treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA 1 mg L-1 + BA 0.1 mgL-1 treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

Development of a new culture medium and efficient protocol for in vitro micropropagation of Ceratonia siliqua L.

A new basal culture medium was developed and tested using a rapid and efficient protocol of in vitro axillary shoot bud proliferation of Ceratonia siliqua L., an important Mediterranean Fabaceae plant species. In a first experiment, the new formulated ‘LA’ mineral composition significantly improved shoot growth and proliferation as compared with Murashige and Skoog medium (MS, 1962) in both solid and liquid culture media. However, the liquid culture system proved to be the most suitable for shoot induction, shoot length (about fourfold higher), and multiplication rate (about two-fold higher), the difference being significant. The measured growth and proliferation parameters were further improved when LA mineral composition was optimized, in a second experiment. The highest multiplication rate (6.3) was achieved during the second subculture using the optimized ‘LAC’ medium. Noticeably, hyperhydricity and shoot-tip necrosis symptoms were absent in both formulated LA and LAC compositions when using the liquid culture system. In vitro rooting in solid medium showed 41.7 to 46.3% response on a solid medium which was more suitable than the liquid culture system, the difference being significant. In contrast, pretreated microcuttings with 3 μM IBA (indole-3-butyric acid) were successfully rooted ex vitro, showing significantly higher response (91.7%), average root number (8.3), and root length (31.5 mm). The plantlets were successfully acclimatized showing more than 90% survivability and normal morphology. The present study is a first cost-effective protocol for carob micropropagation combining the use of the newly formulated LAC basal medium, a liquid culture system, and ex vitro rooting.

Establishment of tissue culture and acclimatization method for in vitro mass propagation of Echeveria laui and Echeveria elegans

The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with 0 ~ 2 mgL-1 NAA and 0 ~ 4 mgL-1 BA. The upper and middle parts of leaves both showed 100% callus formation rate with NAA 1 mgL-1 and BA 1 mgL-1 treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at NAA 2 mgL-1 and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at NAA 0.1 mgL-1 and BA 3 mgL-1 treatment in E. laui and NAA 0.3 mgL-1 in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in NAA 1 mg L-1 and BA 1 mgL-1 treatment in E. laui and 12.0 shoots in most effective NAA 1 mgL-1 and BA 0.1 mg L-1 treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.

Factors affecting the efficiency of in vitro regeneration from seedling-derived nodal explants of Nyctanthes arbor-tristis L. and evaluation of genetic fidelity

An in vitro micropropagation protocol for Nyctanthes arbor-tristis was developed through the culture of seedling-derived nodal explants. The highest direct shoot induction rate, the mean number of shoots per explant and average shoot length was obtained on MS medium supplemented with 5 mg/l 6-benzylaminopurine (BAP). Full-strength MS medium was found better than other concentrations for organogenesis. Their lower or higher concentration from optimum level was inhibitory for shoot regeneration. Among different carbon sources tested for organogenesis, MS medium supplemented with 87.64 mM sucrose was superior to other carbon sources for in vitro morphogenesis. Best rooting of microshoots was achieved on one-quarter strength MS medium supplemented with 2 mg/l indole-3-butyric acid (IBA). Under this treatment, maximum mean number of roots/shoot and mean root length were 9.40 ± 0.70 and 2.47 ± 0.25 cm, respectively. The genetic fidelity among regenerates and donor plant has been confirmed by ISSR markers.

Factors influencing direct shoot regeneration from leaves, petioles, and plantlet roots of triploid hybrid Populus sect. Tacamahaca

Since the generation of full-sib artificial triploid families, rapid clone establishment and genetic improvements have been needed. Here, we report an in vitro method of direct shoot regeneration of a triploid hybrid poplar [(Populus simonii × P. nigra ‘Italica’) × (P. × ‘popularis’)]. Using different randomized block designs, we selected one triploid to evaluate the explant type, optimal concentrations of plant growth regulators and agar, and culture time under light or dark conditions over 60 days. The highest rate of shoot induction, 80.0%, was obtained using Murashige and Skoog (MS) medium supplemented with 0.2 mg/L benzyladenine, 0.04 mg/L naphthaleneacetic acid (NAA), and 5.5 g/L agar for the first 30 days in the dark, then 3 g/L agar for the next 30 days in light. This last medium yielded the best rate of shoot induction (6.32 shoots/explant). These three media were also used to evaluate the influence of the genotypes of the parents and hybrid triploids on regeneration. Two parents and three of the four full-sib triploids were regenerated successfully; different genotypes and explant types significantly affected the rate of shoot induction and average number of shoots. Leaves but not petioles were a suitable explant. One genotype produced the highest rate of shoot induction of 96.67%. Half-strength MS medium supplemented with 0.2 mg/L indole butyric acid and 0.04 mg/L NAA was the most effective for rooting; rooting rate was 96.67%, survival rate of transplants was 73.33%, and rooting frequency surpassed 85% for each genotype. Overall, this in vitro regeneration system will be useful for the propagation and genetic modification of triploid poplars.

In Vitro Culture of Heuchera villosa ‘Caramel’

The procedure for Heuchera villosa ‘Caramel’ propagation was investigated, which involves shoot regeneration, rooting of regenerated shoots, and acclimation of regenerated plantlets. Petioles, as explants, were cultured on MS medium supplemented with 1-naphthylacetic acid (NAA), benzylaminopurine (BA), thidiazuron (TDZ) and callus formed on all media. Shoots were observed to proliferate from callus on media with BA and NAA, whereas no shoots regenerated on media with TDZ and NAA. On media containing 0.5 or 1.0 mg·L−1 BA in combination with NAA, the regenerated shoots showed severe hyperhydricity, whereas on media containing 0.1 mg·L−1 BA in combination with NAA, the regenerated shoots grew normally. The highest shoot induction rate, 90.6%, was obtained on media containing 0.1 mg·L−1 BA and 0.01 mg·L−1 NAA. The effects of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and NAA on rooting of H. villosa ‘Caramel’ was explored. The highest rooting rate (95%) was obtained on 1/2 MS medium containing 0.2 mg·L−1 NAA. In the subsequent acclimation experiments, about 85% of rooted plantlets survived and grew normally.

English

An efficient micropropagation protocol was developed for the medicinal plant Launaea cornuta using green house axillary buds as explants. The best sterility was obtained at 30% (v/v) local bleach (JIK). Maximum shoot induction rate was achieved when axillary buds were cultured on Murashige and Skoog (MS) Media supplemented with 0.5 mg/L of 6-benzylaminopurine (BAP) for 3 weeks. The highest number of shoot multiplication was obtained when induced shoots were culture on MS media supplemented with 0.5 mg/L BAP and 0.2 mg/L NAA for 30 days. The best rooting response with regard to average root length, rooting percentage and number of roots was achieved within 4 weeks of culture of excised shoots on MS media having 0.5 mg/L BAP. Regenerated plants were successfully acclimatized and about 80 to 90% of plantlets survived under ex vitro conditions. About 170 plants were produced from a single nodal bud of L. cornuta after 60 days. A reproducible protocol was established for in vitro propagation of L. cornuta, an important indigenous vegetable with high medicinal value. Key words: Launaea cornuta, tissue culture, micropropagation, axillary buds, tissue culture.

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.

En-masse production of elite clones of Dendrobium crepidatum: A threatened, medicinal orchid used in Traditional Chinese Medicine (TCM)

Orchids are one of the promising medicinal plant families of which Dendrobium crepidatum. Lindl & Paxton figures out prominently because of its multi directional medicinal attributes. In the present report, in vitro regeneration protocol has been developed from the nodal segments of D. crepidatum, to cater sustainable commercial exploitation and conservation needs. Thidiazuron (TDZ) at 3mg/l singly resulted in the response frequency of 55% which could be increased to 97% by incorporating NAA at 0.5mg/l and TDZ at 2mg/l in combination in the medium. Shoot induction rate was further enhanced with the use of polyamines and at 0.8mM putrescine along with 2mg/l TDZ and 0.5mg/l NAA in the medium 11.8 shoots/explant could be obtained. Highest rooting frequency of the shoots was achieved in medium containing 2mg/l IBA. Genetic stability of the acclimatized plants was assessed using Start Codon Targeted (SCoT) polymorphism and inters simple sequence repeats (ISSR). SCoT revealed a total variability of 10% within the micropropagated plants whereas the cumulative ISSR and SCoT data revealed 6.25% clonal variability indicating high genetic fidelity amongst the regenerates. A comprehensive higher yield of the secondary metabolites along with significant higher antioxidant potentials as compared to the mother plant was revealed using DPPH and FRAP assays. An increased regeneration frequency with a comparative higher yield of secondary metabolite and genetic stability reported in the present communication ensure the ingenuity of this clonal propagation protocol developed for D. crepidatum which can be further utilized in the sustainable commercial utilization and conservation of other medicinally important orchid species.

Direct Organogenesis and Plantlet Establishment via Cotyledon Explants in Medicinally IDirect Organogenesis and Plantlet Establishment via Cotyledon Explants in Medicinally Important Herb Silybum marianum (L.)mportant Herb Silybum marianum (L.)

Objectives: The species Silybum marianum (L.) (Asteraceae) commonly known as Milk thistle is medicinally valuable and due to over exploitation the species may become threatened. Hence, we have made an attempt to optimize the protocol for its direct regeneration through cotyledon explants using in vitro culture technology. Methods/Statistical Analysis: For in vitro multiple-shoot induction, cotyledon explants were excised from surfacesterilized seeds and cultured on MS Basal Medium (MBM) enriched with diverse conc. (10.0–20.0 μM) of BAP/KIN alone and also 10.0 μM BAP/KIN in combination with 1.25–5.0 μM NAA/IAA/IBA. The micro-shoots were cut out and cultured on 1/2 strength MBM enriched with diverse concentrations (5.0-20.0 μM) of NAA/IAA/IBA alone and also coupled with 1.25–5.0 μM BAP/KIN. Findings: In this study, superlative rate of shoot induction, maximal Number of Shoots Per Explant (NSPE) and longest shoot-length were achieved on MBM enriched with 10.0 μM BAP + 2.5 μM NAA. Superlative rate of root induction, mean number of roots per shoot and mean root length were documented on MBM supplemented with 15.0 μM NAA + 2.5 μM BAP. Application/Improvement: Being over exploited, the current technique can be operated for accelerated multiplication and conservation of the species. Further, this protocol can also be operated for transfer of novel genes for enhancement and modulation of bioactive molecules production in S. marianum – a medicinally valuable plant.

Rapid in vitro Propagation of Dodonaea viscosa (Sapindaceae) Through Axillary Bud Multiplication and Indirect Organogenesis

Protocols of axillary bud multiplication were established for Dodonaea viscosa (Sapindaceae). Murashig & Skoog’s medium with 0.2 mg/l BAP with 0.01 Naphthalene acetic acid induced high rate of shoot induction and an average of five shoots per node. Subsequent culture enhanced the number of shoots. Callus initiated from the basal cut end explants differentiated in to more than 10 shoots on MS Medium with 0.4mg/l Benzylaminopurine and 0.02mg/l Indole Butric Acid. Eighty per cent of the rooted shoots survived when transferred to greenhouse and subsequently to the field.

PROPAGATION OF Portulaca oleracea L. IN LIQUID MEDIUM: IMPLICATIONS OF PLANT GROWTH REGULATORS IN CULTURE

Portulaca oleracea L. is a medicinal plant, growing in warm and moist regions of north hemisphere of the world. A protocol for in vitro propagation using nodal shoot segments as explants has been outlined. The percent shoot response with shoot induction rate, 6.4 ± 0.7 shoots per explant, was achieved when cultured on agar-gelled Murashige and Skoog (MS) medium containing 2.0 mg/L of BAP (6-benzylaminopurine). The cultures were amplified by passages on MS medium with 1.0 mg/L each BAP and kinetin (Kn). The best shoot amplification (37.5±0.9 shoots per vessel) was achieved by subculturing of in vitro regenerated shoot clumps on liquid MS medium. Shoots regenerated in vitro by both the processes were rooted on ½ strength of MS medium + 2.0 mg/L of indole-3 butyric acid (IBA). Ninety six percent of the shoots rooted in vitro. The in vitro rooted plantlets were hardened under different regimes of temperature and humidity in the greenhouse. The hardened plantlets were transferred to mixture of soil and manure in polybags.

<i>In Vitro</i> Micropropagation of a Valuable Medicinal Plant, <i>Plectranthus amboinicus</i>

The effect of the plant growth regulators benzyl amino purine (BAP), 1-naphthaleneacetic acid (NAA) and kinetin (KIN) on in vitro shoot induction and proliferation of Plectranthus amboinicus was examined. Explants obtained from lateral shoots and apical shoots of P. amboinicus were inoculated on Murashige and Skoog (MS) culture medium supplemented with different concentrations of BAP, NAA and KIN. When the effect of each growth regulator was considered singly, the highest rate of shoot induction (80% of explants producing shoots) and highest number of shoots produced (2.4 shoots per explant) were obtained from lateral shoot explants cultured on MS media supplemented with 3.0 mg/L BAP within 6 - 7 weeks. Better results were obtained using MS medium supplemented with 1 mg/L BAP + 5 mg/L NAA. Shoot proliferation rose to 85%, while 5.7 shoots per explants were recorded. Among the different media tested for rooting, MS medium supplemented with 1.0 mg/L IBA was the most effective for root induction. The quality of the roots obtained was better than that obtained using MS media supplemented with NAA or IAA.

Development of an efficient regeneration and rapid clonal multiplication protocol for three different Morus species using dormant buds as explants

SummaryAn efficient in vitro protocol using dormant buds as explants was standardised for three Morus (mulberry) species: M. alba, M. indica, and M. laevigata. Explants were collected from field-grown plants and cultured on 1.0_ Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of phytohormones. Thidiazuron (TDZ) at 0.1 mg l-1 or benzylaminopurine (BAP) at 1.0 mg l-1, added separately, gave the best rates of shoot initiation and shoot induction, and required less time for bud sprouting in all three species. Basal MS medium resulted in the lowest rate of shoot induction and the longest duration for shoot initiation. Shoot multiplication was induced by a combination of 0.5 mg l-1 BAP, 0.5 mg l-1 kinetin (Kn), and 0.1 mg l-1 indole-3-acetic acid (IAA). Shoot proliferation and the elongation of adventitious shoots were observed using 1.0 mg l-1 BAP plus 0.2 mg l-1 gibberellic acid (GA3). The rooting percentage was highest (90%) on 0.5_ MS medium supplemented with 0.5 mg l-1 indole-3-butyric acid (IBA), which was more effective than rooting (70%) on 1.0 mg l-1 _-naphthaleneacetic acid (NAA). Well-rooted plantlets exhibited normal growth under greenhouse conditions, with a 98% survival rate. Plants derived from this multiple shoot induction protocol showed no apparent differences in phenotype in the greenhouse. This protocol would be of use for large-scale propagation of mulberry to support ex situ conservation.

Improvement of seed germination and in vitro propagation of Bupleurum latissimum Nakai through embryogenesis

Abstract: Bupleurum latissimum Nakai is a critically important endangered plant belonging to the family Apiaceae. Seed germination was promoted by soaking in the dormancy breaker gibberellic acid (GA3) (50% germination as compared with 4% of the control). An optimal temperature for germination of seed previously soaked in GA3 solution was determined by incubation at various temperatures. Seed germination of 31.8% was observed at 25 °C. These results indicate that the seeds of B. latissimum are difficult to germinate, even when treated with GA3. The greatest callus induction (94.8%) was observed in root explants of seedlings grown on MS medium containing a specific concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) (1.0 mg L-1) and N6-benzyladenine (BA) (3.0 mg L-1). Induction of somatic embryo was observed in 78.5% of the root segment cultured on MS medium containing 3.0 mg L-1 2,4-D alone. The highest shoot induction rate was obtained in MS medium containing 30 g L-1 sucrose (number of shoots, 6.8; average length of shoot, 8.8 cm). After acclimation in artificial soil (1:1:1 mixture of peat moss, perlite, and sand) and transfer to the greenhouse, 98% of plantlets survived over 2 months. This in vitro propagation protocol will be very useful for the conservation of this critically endangered plant.

<i>In Vitro</i> Induction and Identification of Tetraploid in Watermelon [<i>Citrullus lanatus</i> (Thunb.) Matsum and Nakai

This research induced tetraploid watermelon through tissue culture. The cotyledons of a diploid mini-watermelon (A7) were treated with different concentrations of colchicine on medium for different time. The autotetraploid plants were identified basing on morphology, determination of the number of chloroplasts in stomatal guard cells and flow cytometry analysis. A stable autotetraploid material was observed. The results showed that tetraploid watermelons could be obtained under different treatments, and the highest tetraploid induction rate was up to 25 %. The most effective way was cutting the proximal cotyledons at the 7th day after sowing, then explants were cultivated on MS medium with 0.1 %(w/v) colchicine for 72 h, the adventitious shoot induction rate was 62.5 %, and multiplication coefficient was 3.6.

Effect of seaweed liquid extracts and plant growth regulators on in vitro mass propagation of brinjal (Solanum melongena L.) through hypocotyl and leaf disc explants

The effect of seaweed liquid extracts (SLEs) made from Gracilaria salicornia, Padina gymnospora, Padina boergesenii, Gelidiella acerosa and plant growth regulators (PGRs) were examined on in vitro mass propagation using hypocotyls and leaf disc explants of brinjal (Solanum melongena L.) cultivar Pusa purple long. For the germination bioassay, seeds germinated with 20–40 % SLEs exhibited enhanced germination. Initially, hypocotyls and leaf discs were cultured on Murashige and Skoog (MS) medium containing 6-benzylaminopurine, zeatin and thidiazuron for shoot induction. The best responding cytokinin, 6-benzylaminopurine, was employed with different auxins (indole-3-acetic acid, indole-3-butyric acid and 1-naphthaleneacetic acid) for shoot proliferation. In a second experiment, all the four SLEs (10–60 %) combined with MS medium were studied for shoot propagation. Augmented shoots transferred to half-strength MS medium and supplemented with auxins and SLEs (10–70 %) individually to induce rooting. In these experiments high rate of shoot induction (96.2 %), proliferation (6 cm) and rooting (95.3 %) was found with 20–40 % of SLEs. Well-matured plantlets were transferred to soil cups, maintained in a growth chamber for a week to control humidity and then shifted to a greenhouse. This study demonstrated that SLEs could serve as an alternative to phytohormones as they were easy to extract and gave quick and high-frequency mass propagation.

In vitro propagation of Caralluma tuberculata and evaluation of antioxidant potential

AbstractPresent study describes rapid in vitro propagation of Caralluma tuberculata, a traditional medicinal plant, and antioxidant potential of calli and plants extracts. The highest callus induction rate (93.3%) with maximum weight of calli 5.2 g was achieved from shoot tip explants on MS medium supplemented with 9.04 μM 2,4-D and 4.44 μM BA. The maximum shoot induction rate (71.1%) with mean number of shoots 3.66 ± 1.53 and 4.6 cm average shoot length was observed on 13.32 μM BA, 4.52 μM 2,4-D and 2.89 μM GA3 appended in MS medium. The developed shoots were best rooted in the presence of 5.07 μM IAA with 3.0 ± 0.15 roots per plantlet. The plants were successfully acclimatized under in vivo conditions. The plants and calli extracts exhibited good antioxidant activities, however, plant extract activities were more pronounced. The phenolic compounds in plant and calli extracts were 0.16% and 0.057%, respectively. While the flavonoids were 0.092% in plant and 0.039% in calli extract. Total Phenolics, flavonoids; DPPH radical scavenging activity and reducing power potential distributed among different fractions depending upon polarity of the solvent. The highest DPPH scavenging activity and reducing power was exhibited by water fractions; 4.95 mg/mL and 0.729 OD at 10 mg/mL, respectively. The micropropagation protocol can be successfully used for large-scale multiplication and conservation of germplasm of this threatened plant. Furthermore, antioxidant value describes importance of this valuable plant as food and medicine.

<b>Plant regeneration and histological study of the somatic embryogenesis of sugarcane (<i>Saccharum</i> spp.) cultivars RB855156 and RB72454</b> - doi: 10.4025/actasciagron.v36i1.16342

The development of an efficient regeneration system is crucial for the genetic transformation of sugarcane. The aims of this study were to regenerate plants from somatic embryos and analyze the origin and division pattern of cells during the different embryonic developmental stages of the sugarcane cultivars RB855156 and RB72454. For both cultivars, the best results for embryogenic callus induction were obtained with explant cultures on culture medium containing 13.5 µM 2,4-dichlorophenoxyacetic acid (2,4-D) followed by callus culture on solid medium with 4.5 µM 2,4-D. A higher rate of shoot induction was observed in RB855156 with the addition of 8.9 and 17.8 µM benzylaminopurine (BAP); this higher rate was also observed in RB72454 with the addition of 17.8 µM BAP. Murashige and Skoog (1962) (MS) medium containing 2.5 and 5.0 µM indolebutyric acid (IBA) for RB855156 and RB72454, respectively, was suitable for rooting. The plantlets were successfully acclimatized and the plant survival rate was 100% for both cultivars. Histological analysis revealed that the shoots in both cultivars originated from somatic embryogenesis

Ιn vitro plant regeneration from leaf explants of the cherry rootstocks CAB-6P, Gisela 6, and MxM 14 using sodium nitroprusside

Conventional multiplication of cherry (Prunus cerasus L.) rootstocks utilizes division, cuttings, and propagation through seed, which are relatively slow and labor intensive and result in genetic variability. Tissue culture, on the other hand, ensures rapid, large-scale, and low-cost production of genetically identical, physiologically uniform, and pathogen-free plants. In the cherry rootstocks CAB-6P, Gisela 6, and MxM 14, sodium nitroprusside (SNP) promoted callus induction, in vitro shoot proliferation, and rooting from leaf explants in a medium containing 17.6 μM benzyladenine and 2.68 μM α-naphthaleneacetic acid. CAB-6P explants treated with 10 μM SNP gave the maximum shoot number (5), whereas 30 μM SNP gave the longest shoots and the greatest shoot induction rate (26.67%). Best rooting was obtained with 50 μM SNP. In Gisela 6 rootstock, the shoot number (10) and shoot length (20.5 mm) were maximal in the control group without plant growth regulators. The shoot induction rate was enhanced (40%) with 40 μM SNP. SNP at 40 μM resulted in root formation, while 30 μM produced the largest callus size, and 10 μM SNP resulted in the maximum callus fresh weight. MxM 14 leaves treated with 30 μM SNP gave the maximum shoot number (3), root number (7.56), and shoot induction rate (40%), whereas 40 μM SNP gave the longest shoots (12 mm) and roots (20 mm). Best results for callus size, callus fresh weight, and callus induction rate (100%) in the CAB-6P and MxM 14 rootstocks were observed with 30 and 40 μM SNP, respectively. Rooted explants with shoots were gradually acclimatized to the external environment with a high survival percentage (85%). An efficient protocol of indirect organogenesis was established for the three cherry rootstocks using SNP.