Shoot Induction Research Articles

Efeito combinado de citocininas na propagação in vitro de três cultivares de morango

ABSTRACT In the in vitro propagation of strawberry plants, techniques have been applied to obtain disease-free plants with high multiplication rates. To improve the efficacy of micropropagation protocols, it is necessary to determine the effect of growth regulators, mainly cytokinins; however, there is no information regarding the combined use of trans zeatin (Zt) and other cytokinins for shoot induction. The objective of this work was to evaluate the effect of Zt combined with 6-benzylaminopurine (BAP), 6-furfurylaminopurine (KIN) or 2-isopentenyladenine (2iP) on the in vitro propagation of three strawberry cultivars. Combinations of cytokinins, Zt and BAP, Zt and KIN, and Zt and 2iP were proposed to evaluate shoot induction. For the acclimatization of the seedlings, two types of substrates were used (Pro-Mix, Pro-Mix plus humus 2:1). The use of Zt and KIN increased the average number of shoots per explant, and the greatest number of leaves and roots was obtained when Zt and 2iP or Zt and BAP were used. The Pro-Mix and Pro-Mix plus humus substrates did not significantly affect the number of leaves or roots, or the SPAD. These results indicate that the use of the combination of Zt and KIN allowed us to obtain the greatest number of shoots per explant that did not produce calli and promoted the development of the root system; likewise, the Pro-Mix or Pro-Mix plus humus substrate were adequate for acclimatization, allowing the growth and development of strawberry plants.

Propagação rápida da planta ornamental Homalomena gigantea usando estacas de rizoma

Abstract This study investigates the critical factors influencing shoot and root regeneration from Homalomena gigantea rhizome cuttings, providing valuable insights for rhizome cutting practices for this plant species. Three key experiments were conducted to assess the impact of plant growth regulator (PGR) concentrations, cutting length, and cutting section on regeneration. In the first experiment, different concentrations of α-NAA and BAP were examined, revealing that PGR selection and concentration play a pivotal role in shoot and root induction and plantlet development. The highest success rates were observed at 150 ppm α-NAA and 100 ppm BAP, although there was a trade-off between the quantity and quality of plantlets, emphasizing the need for a balanced plant regulator selection. The second experiment investigated the influence of rhizome cutting length. Longer cuttings, particularly at 5 and 7 cm, consistently led to better shoot and root development, highlighting the importance of selecting appropriate cutting lengths. In the third experiment, cutting sections (apical, middle, and distal) without PGR concentrations were assessed, with all sections showing high regeneration rates. However, the apical section produced the highest-quality plantlets with superior growth parameters, followed by the middle section, while the distal section exhibited lower effectiveness. In summary, these findings underscore the significance of PGR choice and concentration, cutting length, and cutting section for successful regeneration of H. gigantea from rhizome cuttings. These insights provide valuable guidance for enhancing rhizome cuttings methods for this plant species and promoting efficient propagation.

In vitro regeneration of black pepper (Piper nigrum L.)

This investigation was done to determine how various plant growth regulators such as benzyl adenine (BA) and indole-3-butyric acid (IBA) affected the in vitro plant regeneration of the black pepper plant. The largest number of shoots was produced when the shoot tips and nodal segments of black pepper were utilized as explants and inoculated in Murashige Skoog (MS) media supplemented with 1.5 mg/L BA. The treatment with 2.0 mg/L BA at 8 weeks after induction (WAI) produced the most leaves (3.2). In the 2.0 mg/L treatment of IBA, the greatest number of roots was regenerated. The treatment 2.0 mg/L BA + 2.0 mg/L IBA produced the highest results for shoot induction, shoot length, and overall leaf production when the effects of both hormones were combined. The same treatment also produced the highest proportion of root induction and longest roots. Regenerated plantlets survive at a 46.67% rate in a shaded building and at a 57.14% rate in the open air under bright sunlight. In order to produce black pepper on a big scale, an effective approach for in vitro regeneration of black pepper has been established. Bangladesh J. Agri. 2023, 48(2): 75-85

A multiple shoot induction system for peptide-mediated gene delivery into plastids in Arabidopsis thaliana, Nicotiana benthamiana, and Fragaria×ananassa.

The plastid is a promising target for the production of valuable biomolecules via genetic engineering. We recently developed a plastid-specific gene delivery system for leaves or seedlings using KH-AtOEP34, a functional peptide composed of the polycationic DNA-binding peptide KH and the Arabidopsis thaliana plastid-targeting peptide OEP34. Here, we established a liquid culture system for inducing multiple shoots in the model plants A. thaliana and Nicotiana benthamiana and the crop plant strawberry (Fragaria×ananassa) and tested the use of these plant materials for peptide-mediated gene delivery to plastids. Our liquid culture system efficiently induced multiple shoots that were enriched in meristems. Using these meristems, we performed KH-AtOEP34-mediated gene delivery to plastids and tested the delivery and integration of a cassette composed of the spectinomycin resistance gene aadA, the GFP reporter gene, and sequences homologous to plastid DNA. Genotyping PCR revealed the integration of the cassette DNA into plastid DNA several days after delivery in all three plants. Confocal laser scanning microscopy and immunoblotting confirmed the presence of plasmid-derived GFP in the plastids of meristems, indicating that the plasmid DNA was successfully integrated into plastid DNA and that the cassette was expressed. These results suggest the meristems developed in our liquid culture system are applicable to peptide-mediated delivery of exogeneous DNA into plastids. The multiple shoots generated in our liquid novel culture system represent promising materials for in planta peptide-mediated plastid transformation in combination with spectinomycin selection.

An efficient direct organogenesis protocol for in vitro clonal propagation of Rubia cordifolia L.

This comprehensive study delves into the intricate process of in vitro clonal propagation of R. cordifolia, with a particular focus on optimizing a method of direct organogenesis. The choice of direct organogenesis over indirect organogenesis is driven by the propensity for somaclonal variations associated with the latter. The presented research explores the critical factors influencing direct organogenesis, including the selection of appropriate explants and the optimum use of plant growth regulators. Notably, nodal explants have demonstrated great potential for regeneration, and the choice of cytokinins such as kinetin (KIN) and 2-isopentenyladenine (2-iP) for shoot induction has proven highly effective, with 1.0 mg/L KIN supplementation to MS media emerging as the most favourable choice resulting in 12.14 ± 1.58 shoots and 29.78 ± 1.93 axillary buds. Subsequent adventitious rooting using indole-3 butyric acid (IBA) and indole-3-acetic acid (IAA) resulted in a 100% rooting response. Among the concentrations tested, 1.0 mg/L IBA was found most effective, yielding the best rooting response (22.9 ± 1.06 roots/ micro-shoot). Acclimatization of in vitro cultures was achieved by gradually exposing them to natural conditions, maintaining high humidity and minimizing photorespiration. The morphological transformations observed, including robust leaf development contributed to the high acclimatization success (100% survival). Additionally, the genetic stability was confirmed using DNA-based ISSR markers, with complete monomorphism indicating no variations among the regenerants. The efficient direct organogenesis protocol developed in this study may prove a valuable contribution towards the mass clonal propagation and conservation of R. cordifolia, an industrially and medicinally important plant.

Plant Regeneration via Organogenesis in Jerusalem Artichokes and Comparative Analysis of Endogenous Hormones and Antioxidant Enzymes in Typical and Atypical Shoots.

The Jerusalem artichoke (Helianthus tuberosus) is a tuberous plant with considerable nutrient and bioactive compounds. The optimization of the in vitro clonal propagation protocol is critical for large-scale reproduction and biotechnological applications of Jerusalem artichoke production. In this work, in vitro plant regeneration from the stem nodes of the Jerusalem artichoke via direct organogenesis is presented. In the shoot induction stage, the stem segments produced more shoots with vigorous growth on MS medium containing 0.5 mg/L 6-benzylaminopurine (6-BA). The concentrations of 6-BA and gibberellic acid (GA3) were both optimized at 0.5 mg/L for shoot multiplication, and the combination of 0.05 mg/L indole-3-butyric acid (IBA) and 0.05 mg/L 1-naphthylacetic acid (NAA) was the most responsive for root induction, yielding the largest number of roots. The regenerated plantlets were successfully hardened at a 96% survival rate and vigorously grew in the field. The genetic stability of the regenerated plants was confirmed by flow cytometry and simple sequence repeat (SSR) analysis. However, 17.3% of shoots on the optimum shoot induction medium had withered leaves and excessive callus (atypical shoots), which greatly reduced the induction efficiency. Enzyme activity in the typical and atypical shoots was compared. The atypical shoots had significantly higher levels of endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA), as well as increased activity of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD), whereas the content of 6-BA, zeatin (ZT), and GA3 was significantly reduced. The activity of the three enzymes was positively correlated with the content of IAA and ABA, while being negatively correlated with that of 6-BA, ZT, and GA3. The results suggest that the poor growth of the atypical shoots might be closely related to the significant accumulation of endogenous IAA and ABA, thus significantly increasing antioxidant enzyme activity.

Allele-Specific Transcriptional Regulation of Shoot Regeneration in Hybrid Poplar

Plant tissue regeneration is a key process for genetic transformation and genome editing. The exploration of regulatory mechanisms in plant regeneration would improve regeneration efficiency. In comparison to some model plants, the genomic heterozygosity is much higher in forest trees, increasing the complexity of transcriptional regulation. Here, we report the allele-specific transcriptional analysis in hybrid poplar 84K (Populus alba × P. tremula var. glandulosa cv. 84K) during the shoot regeneration process. Firstly, 180 regeneration-related genes (REGs) and 2446 REG-homologous genes (REGHs) were identified in hybrid poplar. The expression patterns of REGs exhibited that about half of them were positively correlated between poplar and Arabidopsis at the locus level. The expression levels of REGHs vary among the gene family at different stages during callus and shoot induction. Among the gene clusters with similar expression patterns, the distribution of gene families in poplar and Arabidopsis also exhibits large variations. At the allele level, most of the allele pairs of REGs were positively correlated in expression. The expression patterns of genes in auxin synthesis, transport, and signaling pathways agree with the general patterns. Due to the presence/absence of variations between two subgenomes, two YUC alleles and two IAA alleles are present only in one subgenome, and the expression patterns of the two alleles are greatly different. Our analysis indicates the conservativeness and diversity of transcriptional regulation during shoot regeneration in poplar and Arabidopsis. The complexity in allele expression contributed by heterozygosity suggests the importance of genotyping in the screening of explants for plant regeneration.

Establishment of Dendrobium wilsonii Rolfe in vitro regeneration system

Tissue culture processes have been established for Dendrobium wilsonii Rolfe, a valued ornamental and medicinal orchid. Pods aged 180 days served as seed sources. These seeds underwent surface sterilization and were then aseptically germinated on Murashige and Skoog (MS) medium supplemented with 0.5 mg L−1 Benzyl adenine (BA), 0.5 mg L−1 Naphthalene acetic acid (NAA), 20 g L−1 Sucrose (Su), 7 g L−1 Agar, 1 g L−1 Activated charcoal (AC), 1 g L−1 Peptone and 1 g L−1 Hyponex No.1 incubated in a growth chamber at 25.2°C with a 12-hour photoperiod and a light intensity of 2200 lx. Multiple shoot induction and proliferation was on MS medium in vitro produced shoots were rooted on MS medium with Hyponex No.1, 25 g L−1 Su, 1 g L−1 AC and 1.5 g L−1 Peptone. Peptone enhanced seedling growth and promoted clumped shoot development. The establishment of Dendrobium wilsonii Rolfe regeneration system provides a methodology for conserving germplasm resources and for commercial propagules production.

Influence of High Concentrations of Copper Sulfate on In Vitro Adventitious Organogenesis of Cucumis sativus L.

Copper (Cu) is an essential plant micronutrient. This report is the first to assess the effects of high copper sulfate (CuSO4) levels on in vitro callus and shoot formation of cucumber. Four-day-old cotyledon explants from the inbred line Wisconsin 2843 and the commercial cultivars Marketer and Negrito were used. Murashige and Skoog (MS)-derived callus and shoot induction medium containing 0.5 mg L−1 indole-3-acetic acid (IAA) and 2.5 mg L−1 6-benzylaminopurine (BAP) was supplemented with CuSO4 (0.2–5 mg L−1). The response on callus-derived shoots showed that the optimal concentration of CuSO4 was 8- to 200-fold greater than in standard MS medium. Shoot frequency (SF) and shoot number (SN) were assessed, and Marketer > Negrito > Wisconsin 2843, where 1, 0.2 and 5 mg L−1 CuSO4 produced the highest results, respectively. SF and SN increased 6- and 10-fold in Wisconsin 2843 and twice in the other cultivars. All explants formed calluses, and in two of the three cultivars, callus extension was significantly affected by CuSO4 application. SN showed a strong relationship with CuSO4 levels and no association with callus extension. The results show that specific CuSO4 concentrations higher than in standard MS medium increase adventitious cucumber shoot organogenesis.

Effect of graphene oxide nanoparticles on in vitro growth of Fragaria x ananassa (Cameron Highlands white Strawberry) and evaluation of genetic stability using DAMD and ISSR markers

Graphene oxide (GO) is a novel nanomaterial with distinct physical properties and significant biological applications. The use of GO in plant tissue culture offers several new properties and potential applications. This research is vital due to the growing need for innovative techniques to promote plant growth, improve plant productivity and mitigate challenges posed by environmental stressors. This study focused on the rare Cameron Highlands white strawberry plants (Fragaria x ananassa) and addressed issues such as callus production during direct shoot induction and hyperhydricity. The research aimed to investigate the effects of GO on the regeneration process and genetic stability of white strawberry plants and to use molecular markers to ensure that plants propagated in vitro are true to type. For this purpose, shoot tip explants were used and different concentrations of GO (0, 2.5, 5.0, 7.5, 10 mg/L) were added to the Murashige and Skoog (MS) medium for six weeks. The results showed that the optimum concentration for promoting the development of white strawberry seedlings was 7.5 mg/L of GO. The study also revealed that the addition of 7.5 mg/L GO in combination with 8 μM TDZ to the MS medium facilitated the induction of multiple shoots. Moreover, the clonal fidelity of the in vitro plants treated with GO showed a genetic similarity of over 97%. These results confirm that lower GO concentrations improve plant development and stability. Consequently, this nanomaterial has a positive effect on the growth of strawberry plants and is therefore well suited for strawberry tissue culture.

Establishment of In Vitro Cultures and Genetic Fidelity Analysis in Valeriana jatamansi -a High Value Endangered Medicinal Herb

The present study was conducted in the Department of Biotechnology, College of Horticulture and Forestry, Neri, Hamirpur, Himachal Pradesh, India during 2018–2020 for 18 months to develop an efficient, rapid and reproducible protocol for in vitro establishment along with analysing antioxidant activity. In the present study, surface sterilization protocol was standardized using Sodium hypochlorite and mercuric chloride as sterilant. Maximum direct shoot induction frequency from nodal explant was achieved on Murashige and Skoog medium enriched with BAP (5.0 mg l-l), TDZ (3.0 mg l-l) and NAA (1.0 mg l-l) for 18 days. Microshoots inoculated on MS media supplemented with 4.0 mg l-l BAP, 2.0 mg l-l TDZ, 1.0 mg l-l NAA and 0.5 mg l-l GA3) showed maximum frequency of shoot multiplication with average shoot length (4.36±0.08 cm) and average shoot number (14.00±0.57). A 75.00±1.15% rooting with significantly high mean root number (8.00±0.57) and root length was achieved in full strength MS medium, supplemented with same concentration i.e. 4.0 mg l-l BAP, 2.0 mg l-l TDZ, 1.0 mg l-l NAA and 0.5 mg l-l GA3) combination. Maximum plantlets survive after 1 year of acclimatization. RAPD and ISSR markers were used to confirmed genetic stability of in vitro raised plants by showing 100% monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant.

In vitro Propagation of Lagenaria Siceraria: A Plant of the Cucurbitaceae Family with High Medicinal Value

Background and Objective: Lagenaria siceraria is extensively grown as a vegetable crop in tropical and subtropical regions of the world as a source of food and for it’s medicinal potential. The purpose of this research was to determine the optimal conditions for plantlet regeneration of Lagenaria siceraria microshoots on various concentrations of benzyl amino purine and naphthalene acetic acid in vitro.
 Materials and Methods: The research was designed as a completely randomized design. Shoot tips of Lagenaria siceraria were surface sterilized with sodium hypochlorite(NaOCl) at different concentrations (0,3,4,5,6 and 7 mg/L) and cultured on Murashige and Skoog (MS) solidified medium supplemented with 6-benzylaminopurine (BAP) at different concentrations of (0,0.5,1.0,1.5,2.0,2.5 and 3.0mg/L) for shoot induction and the obtained shoots were transferred to MS medium containing naphthaleneacetic Acid(NAA) at different concentrations (0,0.5,1.0,1.5,2.0,2.5 AND 3.0mg/L) for root induction.
 Results: Surface disinfection was optimal after immersion of shoot tips and/or nodal explants in cleaning detergent (5 min), ethanol 70% (1 min), and sodium hypochlorite 5mg/L (15 min). MS medium supplemented with 2.0 mg/L BAP was most effective in inducing shoot multiplication (100%), with an average of 4,33 shoots per explant after 4 weeks. MS medium supplemented with 2.0 mg/L NAA was most appropriate for rooting (100%) with an average of 5.33 roots per shoot after 4 weeks.
 Conclusion: shoots sterilized with 5mg/L NaOCl and inoculated in MS medium supplemented with 2.0 mg/LBAP and 2.0 mg/L NAA were optimal conditions for the surface sterilization, shoot and root induction of Lagenaria siceraria shoots.

Ex vitro shoot induction of rodent tuber (Typhonium flagelliforme) with benzyl amino purine and naphthalene acetic acid

A medicinal plant called rodent tuber has the potential to both prevent and treat various diseases. There is no Standard Operating Procedure (SOP) governing the growth method of rodent tuber because it still grows wild and is rarely cultivated. This research aimed to study the optimal growth of rodent tuber for mass seed propagation based on the type of explants and the concentration of auxin and cytokinin combinations for inducing shoots. While generative parameters (harvesting of tubers/shoots) were observed up to 13 WAP (Week After Planting), vegetative parameters were observed up to 10 WAP. The results showed that the best type of explants for shoot induction and growth of rodent tuber were whole plants with a survival rate of 100%; the number of shoots was 6.65; the plant height was 21.27 cm with 6.34 leaves; 26.56 roots with a length of 16.05 cm; and 40.80% flowering plants. PGR (Plant Growth Regulator) combination of 50 ppm BAP (Benzyl Amino Purine) and 100 ppm NAA (Naphthalene Acetic Acid) became the best explant soaking solution by forming eight shoots and the height of plants was 24.13 cm with seven leaves. The explant produced the highest root length in a combination of 100 ppm BAP and 50 ppm NAA, which was 17.70 cm. The interaction between whole plant explants immersed in 0 ppm BAP and 100 ppm NAA solution resulted in the most significant number of roots (41). Those goals can support the SOP of propagated methods and ensure the consistency, quality, and availability of raw materials for traditional medicine.

Micropropagation of autochthonous olive varieties from Türkiye

Olive (Olea europaea L.) is one of the oldest cultivated fruit species in the world. Fruits and oils of autocthonous olive varieties (native Turkish olive varieties) with unique sensory properties (taste, smell and aroma) gain importance recently. Particularly olive oil companies looking for varieties that have distinct taste, smell and aroma in their oil. Propagation of olive varieties by cuttings and grafting is very difficult and expensive, therefore it is important to find solution for easy and mass propagation. Micropropagation is particularly beneficial to propagate plants that are difficult to reproduce conventionally or to ensure virus-free plants or plants with particular qualities. In this study, in vitro micropropagation success of two autocthonous olive varieties (‘Mavi’ and ‘Guleki’) grown in the origin center of olive, Southeast Anatolia, Türkiye was investigated. Both varieties, have distinct smell, taste and aroma and are difficult to root by cuttings. The effects of three different medium OM (Olive Medium), WPM (Woody Plant Medium) and DKW (Driver-Kuniyuki Walnut Medium) and two different growth regulators BAP (6-Benzylaminopurine) and Zeatin on shoot induction (proliferation) in the in vitro micropropagation of the olive varieties were examined. Obtained shoots were later subjected to in vitro rooting and acclimatization. The highest proliferation efficiency and shoot length for both varieties were obtained with the use of 1 mg Zeatin+0.1 mg GAɜ hormone combinations on OM medium. ‘Mavi’ variety formed more roots compared to ‘Guleki’ (3.33 vs. 2.75 per shoots), and gave the highest rooting rate of 74.33% with 2 mg IBA (Indole Butyric Acid) treatment on ½ OM medium. In terms of rooting rate, Guleki gave the highest rooting (100%) on medium containing ½ OM, 0.2 mg GA3 and 4 mg IBA. 60% and 75% of the micropropagated plants of ‘Mavi’ and ‘Guleki’ varieties adapted well to the external conditions.

Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar.

A series of tissue culture regeneration protocols were conducted on gray poplar (P. tremula × P. alba) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L-1 kinetin (KT) and 1.00 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L-1 thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2-41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L-1 6-benzylaminopurine (6-BA), 0.10 mg L-1 indole butyric acid (IBA) and 0.001 mg L-1 TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L-1 naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L-1 NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L-1 NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2-6 weeks compared to that of the indirect shoot induction pathway.

Plant Regeneration via Somatic Embryogenesis and Indirect Organogenesis in Blue Honeysuckle (Lonicera caerulea L.)

Blue honeysuckle (Lonicera caerulea L.), which belongs to the Caprifoliaceae family, is an emerging fruit crop worldwide. For the development of a transgenic system and multipurpose tissue culture, this study for the first time established an in vitro regeneration system via somatic embryogenesis, as well as improving the previously established indirect organogenesis-based regeneration system. For embryogenesis, Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) showed the highest induction rate of the embryogenic callus (97.6%), and MS supplemented with 0.1 mg/L 6-benzyladenine (6-BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 0.5 g/L activated carbon (AC) achieved the highest somatic embryo rate (28.3%). For indirect organogenesis, MS medium supplemented with 1.0 mg/L 6-BA and 0.1 mg/L NAA resulted in the highest non-embryogenic callus induction rate (98.9%) and adventitious shoot induction rate (51.6%). For adventitious root induction, MS supplemented with 1.0 mg/L indole-3-butyric acid (IBA) achieved the highest root induction rate (96.0%) and average root length (4.6 cm), whereas MS supplemented with 0.5 mg/L indole-3-acetic acid (IAA) resulted in the highest average regenerated root number (8.8). The total time for the regeneration from explants to soil-planted seedlings (10 euphylla) was 105 and 150 days with an efficiency of 44.1% and 23.9% through organogenesis and somatic embryogenesis, respectively. This study provides a powerful tool for rapid propagation, proliferation, and transformation, as well as laying a technological foundation for gene function research and genetic improvement of blue honeysuckle.

Methodology of Micropropagation of Elite Genotype in Lotus (Nelumbo nucifera) Genotype Lakshmi

The lotus (Nelumbo nucifera Gaertn.) is an aquatic plant grown extensively throughout Asia, mostly for commercial and ornamental purposes. It is mostly propagated via rhizomes. A potent and reproducible plant regeneration system for lotus has been developed in this work employing rhizomes, shoot tips from the mother plant, and mature and immature embryos collected from eighteen and twenty-six-day-old aseptically fertilized seed. Shoot induction, number of shoots, and length of roots were induced on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of 6-Benzylaminopurine (BAP) mgL-1 (0.5, 1, 2, 3, 4). The response of the explant varied with different BAP concentrations. The most favorable outcomes were achieved using MS medium treated with 0.5 mgL-1 6-Benzylaminopurine (T2).

In vitro Regeneration of Multiple Shoots in Abelmoschus esculentus (L.) Moench (Okra) via Apical Shoot Meristem Culture

Introduction: To develop the efficient protocol for multiple shoot regeneration of A.esculentus by apical root culture method. Abelmoschus esculentus L., commonly known as okra, is a member of the Malvaceae family, which is widely consumed due to its high nutrient content and potential therapeutic properties. Okra contains various bioactive compounds that ex-hibit antibacterial properties and may be useful in treating type-2 diabetes, digestive diseases, and liver detoxification. To select the plant for the present and prepare the efficient protocol for the development of multiple shoot regeneration culture method. Methods: In this study, we developed an efficient protocol for multiple shoot regeneration of A. esculentus using the apical shoot culture method. Mature shoot apex explants of the germinated A. esculentus genotype CoBhH1 were cultured on Murashige and Skoog (MS) medium supple-mented with different concentrations of cytokinins (BAP and TDZ) and auxins (IAA and NAA) to determine the optimal conditions for shoot induction. Results: The highest number of multiple shoots (27.04 shoots) was obtained with 0.8 mg/L TDZ. Excised shoots were cultured on MS medium supplemented with GA3, which induced elongation of the shoots to a maximum of 8-10 cm. Regenerated plantlets were successfully transferred to soil, with a 100% survival rate and no differences in morphology or growth characteristics com-pared to control plants. Rooting was achieved with 1 mg/L IBA. Conclusion: This study provides an efficient protocol for multiple shoot regeneration of A. esculentus through apical shoot culture, which has potential applications in plant breeding and genetic engineering.

The Effect of PVP (Polivinil Pirolidon) and BAP (6-benzylamino purine) on Shoots Induction of Teak Plus Perhutani (Tectona grandis)

The goal of this study is to see how BAP and PVP concentrations affect in vitro teak shoot induction. The research was carried out at Diponegoro University’s Tissue Culture Laboratory, Faculty of Animal Husbandry and Agriculture, between April and June 2022. The research method was a 4x4 factorial design based on a completely randomized design (CRD) with three replications. The first factor was the concentration of PVP (mg.L-1): 0, 110, 220, and 330, and the second was the concentration of BAP (ppm): 0, 1.3, 2.6, and 3.9. Duncan’s Multiple Range Test (DMRT) and orthogonal polynomials were used to test the data for variance. The interaction between the 0 mg.L-1 PVP treatment and the 110, 220, and 330 mg.L-1 PVP treatments after the addition of 2.6 ppm BAP had the quickest callus emergence time, with an average of 7 days. PVP treatments of 110 mg.L-1 and 220 mg.L-1 had a significant effect on the shoot length parameter. BAP concentrations of 1.3 and 2.6 ppm were found to increase the shoot length, number of shoots, and number of leaves response parameters. The 1.3 ppm BAP treatment also reduced browning by 17% in the explants. There was no effect of PVP or BAP treatments on shoot emergence time, leaf emergence time, or the percentage of explants that formed shoots.

Role of phytohormone on in vitro regeneration of tea [Camellia sinensis (L.) O. Kuntze

Phytohormones are small molecules present in very low concentrations in plants and provide a promising strategy to improve mass culture biotechnology due to their intrinsic role in growth, development, and survival. This study is aimed at evaluating the effect of different plant growth regulators on in vitro callus induction and shoot regeneration in tea. The experiment was conducted at Complete Randomize Design (CRD) with three replications. Callus induction and shoot regeneration was done by using Murashige & Skoog (MS) media supplemented with various concentrations and combinations of Naphthalene Acetic Acid (NAA), Benzyl Adenine (BA), and 2,4-Dichlorophenoxyacetic Acid (2,4-D). Shoot tips and nodal segments were used as explant. The highest percentage (86.51%) of callus induction was observed in the combined treatment of 2.00 milligram per litre (mg/L) BA + 1.50 mg/L 2, 4-D in 2.12 weeks. The same treatment produced the highest weight of calli, which was 0.18, 0.56, and 1.11 grams (g) for 6, 10, and 14 weeks after inoculation (WAI), respectively. The highest percentage (84.39%) of shoot induction was recorded with 2.00 mg/L BA + 2.00 mg/L NAA in 32.43 days. The maximum number (1.42, 2.46, and 4.49) and the highest length of the shoot (1.41, 3.28, and 4.61 cm) at 60, 90, and 120 DAI (days after inoculation) were also recorded from the same treatment. No callus or shoot induction was recorded in the controlled treatment. The protocol developed from the present study might be useful for the large-scale production of healthy, disease-free planting material for tea.