Number Of Plantlets Research Articles

Effect of Organic Substrates on Macro-propagation of Banana under Island Conditions

Banana is an important tropical fruit crop grown globally and is the world’s most important agricultural food commodity. Plantation crops take a major share of cultivated area in Andaman and Nicobar Islands. Banana is one of the important fruit crops that is highly suitable to grow as intercrop in plantation based cropping system. Local varieties of banana like Korangi, Mitta Champa, Khatta Champa and Cheenakela are cultivated by the farmers traditionally to meet the local demand. However, in recent past growing commercial varieties of banana has become popular in the Island due to consumer demand. Traditionally banana is propagated by suckers and wide variability was observed in sucker production among the commercial varieties. Due to this large scale multiplication of banana through suckers could not meet the planting material demand in the Island. Hence, a study was undertaken during 2020-2022 to standardize the macro-propagation technology using different organic substrates in the commercial variety ‘Poovan’. Nine different organic substrate combinations were used in the present study to enhance the efficacy of lateral bud development and plantlet production. Suckers weighing 1.0 to 1.5 kg were used as the propagule. Among all the organic media combinations tested, significant differences were observed in terms of plantlet production, root and shoot growth. The treatment T8 (Coircompost: sawdust: FYM: vermicompost (1:1:1:1) with Arka microbial consortia @10 kg) produced maximum number of primary (5.60) and secondary buds (12.07) during primary and secondary capitation of decorticated mother rhizome. Early bud emergence (11.5 days), maximum number of plantlets (12-13 per mother rhizome), maximum shoot length (46.43 cm), maximum shoot girth (3.03 cm) and maximum number of leaves (4.50) were also observed in the treatment T 8. Minimum number of days was taken (52.70 days) for plantlet separation during secondary capitation in the same treatment T 8 suggesting that the treatment combination (Coircompost: sawdust: FYM: vermicompost (1:1:1:1) with Arka microbial consortia @10 kg) was effective for macro-propagation of banana Cv. Poovan under Island ecosystem.

Effect of salt stress on plant regeneration efficiency in primed and non-primed seed's calli of rice (Oryza sativa L.) variety Swarna.

This study leads with the primed seeds of rice (var. Swarna) with distilled water (D.W.) and various concentrations of Mg(NO3)2 (0-8mM)/Kinetin (0-5ppm) alone or in combination with screen out the regeneration medium induced tolerance level of NaCl. To fulfill the objective, the primed and non-primed rice seeds were inoculated in MS medium supplemented with 30gL-1 maltose + 1gL-1 casein hydrolysate and 2mgL-1 of 2,4-D for callus induction and cultured up to 45days in two sets: one set for regeneration purpose in NaCl-induced regeneration medium and another set was used to study the physiological potentiality of the callus. The 45-day-old calli were transferred into regeneration medium MSR (MS medium for regeneration) (BAP: NAA: Kinetin = 4:1:1) containing NaCl with a concentration range of 0 to 300mM. The number of regenerating calli and shoot regeneration percentage, number of plantlets obtained from one callus, recovery of plantlets from each concentration of NaCl and proline estimation from the leaf of the regenerated plantlets were determined from one set obtained after 45days. The calli obtained from another set after 45days, the frequencies of total and embryogenic calli induction percentage, fresh and dry weights, proline content, nitrate reductase and superoxide dismutase activities were measured. The calli obtained from 2.5ppm kinetin + 4mMMg(NO3)2 primed seeds were showed best result as compared to the other treatments for the above-mentioned parameters in different concentrations of NaCl-induced medium and survive up to 200mM concentrations of NaCl.

Nano-Integrated Plant Tissue Culture to Increase the Rate of Callus Induction, Growth, and Curcuminoid Production in Curcuma longa.

Turmeric has attracted a significant amount of interest in recent years due to its strong antimicrobial properties. The tissue culture of turmeric is preferred to obtain disease-free, highest number of plantlets with good uniform chemistry. However, there is a need to increase the speed of the whole process to meet the growing demand for planting materials and to save time and resources. Iron oxide nanoparticles (Fe3O4 NPs) showed positive effects on callus initiation time, proliferation rate, percent root response, shoot length, percent rooting, and number of roots per explant. Highest callus induction, i.e., 80%, was recorded in cultures that were grown in the presence of 15 mg/L of Fe3O4 NPs. Callus initiated earlier in culture tubes that received green synthesized iron nanoparticles in a concentration between 10-15 mg/L. Biofabricated nanoparticles were characterized for their size, physiochemical, and optical properties through UV-Vis spectroscopy, FTIR, XRD, and SEM. Curcuminoids profiling was performed by implementing LC-Ms that revealed increased quantities in plantlets grown in nano-supplemented media when compared to the control.

The Influence of Maturity, Storage, and Embryo Size on Coconut Callus Induction Success

Coconut palms (Cocos nucifera L.) are globally significant palms with both economic and cultural value. Despite the increasing demand for coconut products, production is decreasing globally due to palm senility, pests, and diseases. It has been estimated that over half of the world’s coconut palms need to be replaced immediately. The coconut industry has acknowledged that conventional propagation methods are unlikely to yield sufficient high-quality planting material. Therefore, coconut tissue culture is considered a potential solution to this problem. By using coconut tissue culture, a large number of plantlets can be obtained in a short period of time. In this study, the quality of explants and the development stage (visible shoot/non-visible shoot) of coconut used for micropropagation were examined. To our knowledge, little research has been undertaken on this aspect of coconut micropropagation. Our results indicated that tender coconut fruit exhibited an advantage over mature fruits. In addition, coconut plumule explants subjected to an extended storage of 15 days demonstrated enhanced development compared to those without storage. Notably, smaller embryos utilized as explants displayed superior callus formation compared to their larger counterparts. Finally, embryos possessing shoots exhibited improved callus initiation, albeit accompanied by a more pronounced browning effect. Further investigations are required to obtain more knowledge about the most suitable conditions for plumule explants that lead to optimal callus initiation.

Harnessing the potential of transverse thin cell layer culture for high-frequency micropropagation of Thai ginseng (Kaempferia parviflora Wall. Ex Baker)

In vitro plant propagation techniques play an important role in modern agriculture and horticulture, offering efficient methods for the accelerated multiplication of high-value plant species. Thin cell layer (TCL) culture technique has emerged as a promising approach for large-scale in vitro plantlets production. This study explores the potential of TCL culture as a cutting-edge strategy to enhance the efficiency of multiple shoots formation in Kaempferia parviflora. An attempt was also made to look into the synergistic effects of cytokinin and auxin [6-benzyladenine (BA) - indole-3-acetic acid (IAA)] during micropropagation. Shoot tip explants (from rhizome sprouts) were cultured onto Murashige and Skoog (MS) medium enriched with BA (2.0–8.0 mg/L) and IAA (0.2–0.8 mg/L) in combination where the highest shoot number (7.3 shoots/explant) and length (5.9 cm) was obtained in 8.0 mg/L BA and 0.8 mg/L IAA. Subsequently, the transverse thin cell layer (tTCL) culture system was successfully utilized whereby the shoot tip explants were individually sliced from tips at the meristem and cultured onto MS medium supplemented with BA (2.0–8.0 mg/L) and IAA (0.2–0.8 mg/L) in combination. The regenerated tTCLs exhibited the highest shoot multiplication (9.3 shoots/tTCL; 28 shoots/shoot tip) and length (5.3 cm) in 4.0 mg/L BA + 0.8 mg/L IAA, thus significantly increased number of plantlets were obtained from tTCL culture (953.0 plantlets) compared to shoot tip culture (527.7 plantlets) after six sub-culture cycles. The tTCL regenerants were successfully acclimatized (with 90% survival) and displayed active growth. Genetic fidelity assessment via inter-simple sequence repeats, conserved DNA-derived polymorphism, directed amplification of minisatellite-region DNA, and start codon targeted polymorphism marker systems suggested that a very low level (5.8%; 148 out of 2572 scorable bands) of polymorphism was detected whereas the ploidy level remained unchanged, confirmed by flow cytometry analysis. The implementation of tTCL culture, coupled with a controlled sub-culture regimen, holds significant potential for the efficient commercial-scale production of K. parviflora.

Biochemical and Molecular Characterization of Musa sp. Cultured in Temporary Immersion Bioreactor

The genus Musa sp. contains commercially important fleshy fruit-producing plants, including plantains and bananas, with a strong potential for providing food security and sources of revenue to farmers. Concerns with the quality of vegetative tissues along with the possibility of the transmission of phytopathogens makes the availability of healthy plantlets limited for farmers. Micropropagation of plantains offers an alternative to producing large numbers of plantlets. However, conventional methods of micropropagation have high production costs and are labor-intensive. Recently, the temporary immersion bioreactor (TIB) has emerged as an alternative to conventional micropropagation (CM) methods. Our work utilized SEM microscopy (scanning electron microscope) and molecular and biochemical tools (qRT-PCR and ICP-OES) to characterize and compare the morphological properties, elemental composition, and photosynthetic gene expression of plantains cultured on TIB. Additionally, morphological features of growth and propagation rates were analyzed to compare outputs obtained from TIB and CM. Results showed higher growth and multiplication rates for plantlets cultivated in TIB. Gene expression analysis of selected photosynthetic genes demonstrated high transcript abundance of phosphoenolpyruvate carboxylase (PEPC) in plantain tissues obtained by TIB. Elemental composition analysis showed higher content of iron in plantains grown in TIB, suggesting a potential correlation with PEPC expression. These results demonstrate that micropropagation of Musa sp. via the liquid medium in TIB is an efficient and low-cost approach in comparison with solid media in CM.

Somatic embryo formation, plantlet vigorous and greenhouse preliminary test of garlic bulblet obtained through somatic embryogenesis

Garlic production in Indonesia is still low due to limited availability of seedling. Approximately 10% of garlic yield are used for the next cultivation. Therefore about 90 % of garlic consumption are still imported from China. In addition to conventional seed production, alternative routes are needed, such as through somatic embryogenesis. Research consists of three activities, namely 1) induction and multiplication of callus, 2) embryo somatic and plantlet formation and 3) preliminary test of bulblet in vitro in the greenhouse. Parameters observed in the laboratory activities were time of callus emergence, callus weight and morphology, time of somatic embryo emergence, embryo diameter and weight per bottle, plantlet number and vigorous and bulblet number per bottle, while in the greenhouse, parameter observed were survival of bulblet number, plant height, leaf number per plant, and clove number and weight. The result showed that application of 2,4-D 3 mg/l with casein hydrolysate acid was suitable for callus induction of garlic var. Lumbu Kuning, while BAP 2 mg/l was the most suitable formula for callus multiplication, the addition of paclobutrazol 1 ppm and sucrose 20% in MS medium was the most suitable treatment for plantlet vigorous, while the bulblet formation could be developed by using jasmonic acid with sucrose of 12%. The best medium for growjng media of in vitro bulblet in the greenhouse was the combination of sandy soil, cocopeat and manure.

Investigation of the Influence of Polyamines on Mature Embryo Culture and DNA Methylation of Wheat (Triticum aestivum L.) Using the Machine Learning Algorithm Method.

Numerous factors can impact the efficiency of callus formation and in vitro regeneration in wheat cultures through the introduction of exogenous polyamines (PAs). The present study aimed to investigate in vitro plant regeneration and DNA methylation patterns utilizing the inter-primer binding site (iPBS) retrotransposon and coupled restriction enzyme digestion-iPBS (CRED-iPBS) methods in wheat. This investigation involved the application of distinct types of PAs (Put: putrescine, Spd: spermidine, and Spm: spermine) at varying concentrations (0, 0.5, 1, and 1.5 mM). The subsequent outcomes were subjected to predictive modeling using diverse machine learning (ML) algorithms. Based on the specific polyamine type and concentration utilized, the results indicated that 1 mM Put and Spd were the most favorable PAs for supporting endosperm-associated mature embryos. Employing an epigenetic approach, Put at concentrations of 0.5 and 1.5 mM exhibited the highest levels of genomic template stability (GTS) (73.9%). Elevated Spd levels correlated with DNA hypermethylation while reduced Spm levels were linked to DNA hypomethylation. The in vitro and epigenetic characteristics were predicted using ML techniques such as the support vector machine (SVM), extreme gradient boosting (XGBoost), and random forest (RF) models. These models were employed to establish relationships between input variables (PAs, concentration, GTS rates, Msp I polymorphism, and Hpa II polymorphism) and output parameters (in vitro measurements). This comparative analysis aimed to evaluate the performance of the models and interpret the generated data. The outcomes demonstrated that the XGBoost method exhibited the highest performance scores for callus induction (CI%), regeneration efficiency (RE), and the number of plantlets (NP), with R2 scores explaining 38.3%, 73.8%, and 85.3% of the variances, respectively. Additionally, the RF algorithm explained 41.5% of the total variance and showcased superior efficacy in terms of embryogenic callus induction (ECI%). Furthermore, the SVM model, which provided the most robust statistics for responding embryogenic calluses (RECs%), yielded an R2 value of 84.1%, signifying its ability to account for a substantial portion of the total variance present in the data. In summary, this study exemplifies the application of diverse ML models to the cultivation of mature wheat embryos in the presence of various exogenous PAs and concentrations. Additionally, it explores the impact of polymorphic variations in the CRED-iPBS profile and DNA methylation on epigenetic changes, thereby contributing to a comprehensive understanding of these regulatory mechanisms.

In Vitro Regeneration Protocol for Securidaca longepedunculata Fresen., a Threatened Medicinal Plant within the Region of Lubumbashi (Democratic Republic of the Congo)

Securidaca longepedunculata Fresen. is an overexploited forest species in the Lubumbashi region (south-eastern DR Congo), as its roots are highly valued in traditional medicine. Conventional propagation of this species is affected by seed dormancy and a high mortality rate during early seedling development. To improve on existing methods, we developed an in vitro seed germination protocol. After observing the germination rates, the effects of different doses (0.5, 1, 1.5, and 2 mg/L) of cytokinins (6-benzylaminopurine, kinetin, and meta-topolin) on S. longepedunculata seedling development were compared. Our results showed that soaking for 10 min in NaOCl (10%) followed by 5 min in ethanol (70%) effectively reduced the death rate of seeds while increasing the germination rate to almost 77%. The addition of cytokinins improved plantlet growth: a 12.2× increase in the number of plantlets was obtained with 1.5 mg/L meta-topolin, while only a single stem was obtained from the control. The effects of different auxin types on rhizogenesis did not differ significantly. The best recovery and rooting were noted with microcuttings from the basal parts of S. longepedunculata plantlets. Finally, the seedlings produced survived during the acclimatisation phase regardless of the type of substrate used. The established protocol provides a means for large-scale production of S. longepedunculata plantlets for the restoration of degraded landscapes and agroforestry.

Protocol for plant regeneration from encapsulated somatic embryos of Citrus sinensis L.

BackgroundCitrus is an important fruit crop worldwide; artificial seeds are through encapsulation techniques of somatic embryos having many applications such as large-scale propagation and germplasm conservation. The aim of this study is the investigation of the viability of encapsulated citrus somatic embryos after different storage periods and to convert them into plantlets.ResultsCotyledonary-stage somatic embryos (5–7-mm size) regenerated from stigma explants of Washington navel orange (Citrus sinensis L.) were encapsulated individually in 3% sodium alginate. After different preservation periods (1, 2 or 3 months) at 10 °C, the encapsulated somatic embryos were cultured on Murashige and Skooge (MS) medium solidified with 7 g/l agar and supplemented with 50 g/l sucrose for germination. Percentage of regrowth, germination percentage, number of plantlets, plantlets height and number of leaves/shoot were recorded after different recovery periods. The germination percentages of encapsulated embryos were 90, 62.5 and 40% with storage for one, two and three months, respectively. Encapsulated somatic embryos preserved for 1 month developed the highest number of plantlets, while those preserved for 2 months developed the highest length of plantlets and the highest number of leaves/shoot after a recovery period of 10 months. Molecular analysis was performed of plantlets recovered from somatic embryos after preservation by encapsulation, and the results showed that the percentages of polymorphism were 7.7% with the two primers in all treatments.ConclusionsIt could be concluded that callus developed from stigma explants was able to regenerate indirect somatic embryogenesis after 3 months. Cotyledonary-stage somatic embryos of citrus were successfully preserved by encapsulation using 3% sodium alginate. Frequencies of germination of encapsulated somatic embryos increased with increasing the recovery period on the germination medium. A maximum recovery frequency of 60.8% was obtained from encapsulated somatic embryos cultured on germination medium for up to 10 months. Also, recovery frequency of 62.5% was noticed from encapsulated somatic embryos preserved for 2 months at 10 °C. In addition, results indicated that recovered somatic embryos obtained from encapsulated somatic embryos were able to convert to normal plantlets.

Overview of In vitro Strategies to Conserve Rare and Endangered Plant Species in India

India is one of the seventeen mega-diverse countries in the world. The maximum diversity in India has been observed in the Eastern Himalaya, Eastern part of Western Himalaya and the Western Ghats with its endemism. However, due to intrinsic and extrinsic factors, there has been a reduction in the number of species that are threatened, rare and endangered. Therefore, to sustain the populations of these species, immediate action of conservation is required. Biotechnological approaches have been recognized as an efficient method to conserve the rare and endangered plant species. In the present study, a Google search was carried out along with other scientific databases such as PubMed, SciFinder, Web of Science, and Google Scholar to accomplish a scientific literature search and analysis. In this review, we have described a few potential in vitro methods to protect rare and endangered flowering plants such as tissue culture using different explants, nuclear-DNA assay, cold storage techniques and gene banks. These various strategies are known to have a significant role in the production of a number of plantlets with their quality traits which considerably shows the scope to accomplish the research gap.

Conservation of Threatened Medicinal Plants – A Futuristic Approach

Many of the important plant species are going to be threatened due to various natural as well as artificial reasons. The conservation of medicinal plants such as threatened species can be tackled by scientific techniques such as in situ conservation and ex situ conservation. Tissue culture technology gave us an opportunity to conserve them by achieving multiple numbers of plantlets in small space and less time. The present research paper gives us an opportunity to various methodologies for conservation of these valuable plants for future generations.

Improving Runner Production, Total yield and Postharvest Quality of Strawberry Fruits using Plant Growth Regulators

In this study, different plant growth regulators were tested to improve runner production, total yield and postharvestquality of strawberry fruits. For this, different concentrations of gibberellic acid (GA3) (50 ppm, 100 ppm), naphthaleneacetic acid (NAA) (40 ppm, 80 ppm) and benzyl adenine (BA) (150 ppm, 300 ppm) were used as foliar sprays along withonly water as control. Experiment was arranged in Randomized Complete Block Design (RCBD) with three replicationsand data collected was analysed and means were compared. Results showed that maximum survival percentage (100.00),number of runners per plant (18.77), number of plantlet on each runner (6.88), leaf area (58.45 cm2), chlorophyll contentof leaves (67.87 g/ml), , average number of flowers per plant (28.98), average number of fruits per plant (26.78), fruityield per plant (309.78 g), weight of fruit (11.57 g) and size of fruit (15.34 mm) were found in strawberry plants whichwere sprayed with GA3 @ 100 ppm. However, maximum number of days to flowering (55.89) and number of days toharvest (85.76) were observed in strawberry plants which were treated with only water (control). Postharvest qualityparameters like total soluble solids (TSS) (13.21 °Brix), titratable acidity (1.24%), vitamin C (39.12 mg/100 g), ashcontent (0.78), total carotenoids (4.92 mg/100 g FW), total anthocyanins (4.05 mg/100 g FW), total flavonoids (9.09mg/100 g FW), total phenolics (9.12 mg gallic acid/100 g) and antioxidant activity (102.91 μg/100 mg FW) were alsorecorded maximum in plants, sprayed with GA3 @ 100 ppm. Based on current findings it can be concluded that GA3 @100 ppm was found the best plants growth regulator for vegetative growth and postharvest quality attributes of strawberryas compared to all the other growth regulators tested.

Quantification of endogenous phytochemicals and determination of their exogenous effects in somatic embryogenesis pathways of white and water yams

In vitro propagation of yam via organogenesis is constrained with low multiplication rate. Somatic embryogenesis (SE) has shown rapid multiplication potentials in yam. However, it has not been adopted by practical seed system scenarios due to genotype specificity. Reports have shown that SE is regulated endogenously by phytochemicals, but this is yet to be elucidated for yam. This study identified, quantified endogenous, and evaluated effects of exogenous application of selected identified phytochemicals in yam SE. Callus was induced from in vitro axillary bud explants of three Dioscorea rotundata genotypes in Murashige and Skoog (MS) medium containing 9.1 µM 2,4-dichlorophenoxyacetic acid and 5.4 µM naphthaleneacetic acid. Plantlets were regenerated using MS medium containing 4.4 µM benzylaminopurine and 34.0 µM uniconazole-P. Endogenous phytochemicals associated with axillary bud, calluses, and plantlets were identified and quantified using GC/MS. Effect of selected identified phytochemicals on the genotypes was investigated in a 5 × 6 factorial in completely randomized design (r = 3). Data taken on plantlet regeneration was analyzed using ANOVA at α0.05. A total of 27, 22, and 35 phytochemicals were identified in Kpamyo, Ekiti2a, and Asiedu, respectively. Hexamethylcyclotrisiloxane (36.4%, Kpamyo), Tris-tert-butyldimethylsilyloxy-arsane (59.3%, Ekiti2a), and 4-methyl-2-trimethylsililoxy-acetophenone (52.7%, Asiedu) were highest in callus. N-Methyl-1-adamantaneacetamide (31.8%, Kpamyo) and Tris-tert-butyldimethylsilyloxy-arsane (52.7%, Ekiti2a, Asiedu) were highest in plantlets while Tris-tert-butyldimethylsilyloxy-arsane (41.2%, Kpamyo), hexamethylcyclotrisiloxane (55.8%, Ekiti2a), and erythro-9,10-dibromopentacosane (38.9%, Asiedu) were highest in axillary bud. Plantlet regeneration differed significantly among phytochemicals and ranged from 0.7 ± 0.3 (control) to 4.5 ± 0.5 (40.5 µM phenylacetic acid). Also, genotype × phytochemical interactions on number of plantlets regenerated were significant, and mean values ranged from 0.0 ± 0.0 (TDa2014, 4.8 µM decamethyltetrasiloxane) to 7.0 ± 1.7 (TDa2014, 40.5 µM phenylacetic acid). The application of 40.5 µM phenylacetic acid enhanced plantlet regeneration in Kpamyo and TDa2014 by 5.39% and 343.04%, respectively.

Effect of plant growth regulator on tissue culture media for trigger of Kepok banana

Kepok banana (Musa acuminata x Musa balbisiana) can be grown in a wide area of Indonesia and is still cultivated regularly in rural areas. The need for commercial-quality of Kepok banana seedlings is still an obstacle to supporting export success. Seed propagation using micropropagation techniques is an alternative to be developed in order to meet the needs of appropriate, high-quality, and potential seeds for high productivity. The use of growth regulators and suitable growth media will promote the success of in vitro culture. The method used is the media Murashige-Skoog (MS). The design used was completely randomized and arranged by factorial. Factor 1 is a type of growth regulator with a concentration of 2 mg/L. Factor I is Z1 = Gibberellins; Z2 = IAA; and Z3 = Auxin. Factor II is 50 mg of the growth medium, namely: M1 = Agar medium; M2= Potato medium; M3 = Medium with coconut; M4 = Media Aloe vera. Data analysis used F 5% test and 5% BNT test. The use of a gibberellin growth regulator showed a better shoot formation time of 8 days, the number of shoots was 13.24 seeds, the length of the shoot was 1.25 cm, the root length was 1.52 cm, and the number of shoots was 77.48%. The growing medium using potatoes and young coconut showed a root length of 0.43 cm and a root diameter of 0.34 mm. Young coconut media showed better results in the growth of the number of plantlet shoots as much as 12.53%. The function of growth regulators, gibberellins, IAA, and auxin gave a rapid response to plantlet meristematic changes and shoot formation. While the use of agar, potato, young coconut, and aloe vera growing media gave a slow response to plantlet meristematic changes and the formation of Kepok banana shoots.

Influences of Organic Additives on Asymbiotic Seed Germination of Dendrobium cruentum Rchb. f. for In Vitro Micropropagation

Dendrobium cruentum Rchb. f., a commercial orchid in Thailand, faces the risk of extinction and should be conserved, and propagated for commercial. Therefore, the aim of this research was to identify a suitable VW medium supplemented with organic additives for the micropropagation of D. cruentum through asymbiotic seed germination. The VW media supplemented with separate organic additives (coconut water (CW) (100 mL/L), ‘Hom Thong’ banana homogenate (BH) (1000 mg/L), and potato homogenate (PH) (1000 mg/L) or its combination was determined, and the seed germination rate (%), protocorm formation, and plantlet development were observed. The results showed that a combination of BH and PH resulted in the highest seed germination (94.69 ± 1.55 %) when compared to basal VW (control). Protocorm formation was highest with CW alone (47.38 ± 7.17 %) and a combination of CW and PH (52.13 ± 6.20 %). Additionally, after 10 weeks of culture, supplementation with BH yielded the highest number of plantlets. However, after 24 weeks of culture, each organic additives gave the different developments of healthy plantlets. To investigate suitable potting media for acclimatization of well-grown plantlets in greenhouse conditions, 2 pieces of foam, dried coconut husk, and a combination of peat moss: Sphagnum moss: Pumice: Perlite (3:1:1:1) were investigated. The highest survival rate percentage was found in foam-planted plantlets (80 %). Ultimately, the effectively simple mediums containing organic additives and a one-step protocol were provided for the micropropagation of D. cruentum via seeds, and the derived information could be used for D. cruentum conservation and production. HIGHLIGHTS The effects of organic additives on asymbiotic seed germination for in vitro micropropagation of Dendrobium cruentum Rchb. f. was firstly reported The media supplemented with organic additives yield significantly the highest seed germination, protocorm formation, and plantlet formation and development of D. cruentum when compared to basal media In greenhouse conditions, 2 pieces of foam gave the highest survival rate percentage of plantlets (80 %) The practical and beneficial media supplemented with organic additives for micropropagation of D. cruentum was provided GRAPHICAL ABSTRACT

Influence of plant growth regulators on macro propagation in different banana genotypes

In vivo macro-propagation of banana has gained its momentum as an alternative source for mass multiplication of quality planting materials following some principles like decapitation, decortication of apical meristem of corm and hardening. 6- Benzylamino purine (BAP) @ 40ppm and 60ppm and Indole-3-butyric acid (IBA) @1000 ppm and 2000 ppm were used for locally grown banana genotypes viz. Bharatmani, Chini champa and Bhootmanohar to assess the response of plant growth regulators on shoot and root proliferation. The diseases free uniform corm and sterilized sawdust as uniform rooting media were used in experiment under polyhouse condition. The apical meristem of corm was scooped out with sterilized knife to make a cavity. The growth regulators BAP and IBA) were injected into the cavity of apical meristem 2 days after planting. Proliferation of primary (3.33 and 3.67) and secondary (4.0 and 4.33) buds were noticed significantly by Bharatmani and Chini champa treated with BAP @ 40 ppm and also took less time for bud proliferation.The genotype Chini Champa treated with BAP @ 40 ppm generated maximum number of plantlets per corm (14.67) while the highest number of explants (13.66) per corm after detaching was obtained by Bharatmani using BAP @ 40 ppm.

High-efficiency somatic embryogenesis and plant regeneration in brinjal (Solanum melongena L.) var. Mattu Gulla

ABSTRACT The organogenesis protocol was established for a unique brinjal (Solanum melongena L.) variety, “Mattu Gulla”, which has a distinct size, color and flavor. The objective of this study was to develop a reliable and efficient protocol for the initiation and maturation of somatic embryos into plantlets. The explants were cultured on Murashige and Skoog medium augmented with 2,4-dichlorophenoxyacetic acid (2,4-D) (1.0 mg/L) and benzyl aminopurine (BAP) (1.5 mg/L) for callus initiation. The developing calli (45-days old) were transferred onto an MS medium augmented with plant growth regulators to determine the embryogenic potential of the explants. The proportion of embryogenic callus was higher in hypocotyl-derived calli (HC) with indole butyric acid (IBA) (1.5 mg/L) and BAP (1.0 mg/L with 2,4-D and thidiazuron (TDZ) (1.0 mg/L) than in cotyledon and leaf-derived calli. The embryo initiation was recorded on the 23rd day after subculture from HC with the 2,4-D and TDZ (1.0 mg/L) combination, and germination was recorded. The somatic embryos developed from cotyledon-derived calli showed the highest number of plantlets and a significant percentage of ex vitro survival. In contrast, the lowest number of plantlets was noted from the embryos of hypocotyl-derived calli and ex vitro survival of plantlets. Under greenhouse conditions, the acclimatized plantlets thrived and produced fruits with viable seeds. The established protocol in this study should serve as a platform for large-scale somatic embryogenesis and micropropagation of plantlets. The somatic embryo-based manipulation should be exploited as a biotechnological tool in crop breeding and improving desired agronomic traits.

Rhizome Weight and Number of Sectioning per Rhizome Determine Plantlet Growth and Propagation Rate of Hemerocallis citrina Baroni in Cutting Propagation

Rhizome cutting is prevalent in propagation for Hemerocallis citrina Baroni. This study aimed to reveal the influence of rhizome weight on plantlet growth, and evaluate a new cutting protocol. Three levels of rhizome weight (big (BR), medium (MR) and small (SR)) were compared by measuring plantlet growth four times during cutting propagation. Sectioning rhizomes longitudinally along shaft axis of crown into 2, 3 or 4 parts (S2, S3 and S4), was tested on rhizomes with two bud sizes (Large bud (LB), Small bud (SB)). Propagation coefficient, the number of plantlets obtained per rhizome, kept similar regardless of rhizome weight, while S3 and S4 increased propagation coefficient. Rhizome weight changed the relative growth rates on different dates. SR resulted in lower aboveground dry biomass, leaf area, total fibrous root length, relative total fibrous root length growth rate and N deficiency compared to BR. Sucrose concentration in rhizome decreased with time and fructose concentration was lowest in MR at the end of experiment. Aboveground dry biomass and total fibrous root length were lowest in S4, followed by S3 and S2. Rhizome weight and number of sectioning per rhizome influence cutting propagation, and sectioning rhizomes into three pieces has higher propagation coefficient and less retardation on growth. According to N deficiency in plantlets during the propagation, N fertiliser is probably needed during cutting.

Determination of Optimal Doses of Plant Growth Regulators for In vitro Propagation of Four Potato Varieties (Solanum tuberosum) in Niger

Aims: The main limitation of the production of potatoes (Solanum tuberosum L.) in Niger is the external dependence on the supply of quality seeds. In the context of a national potato seed system, one of the critical phases is the rapid in vitro multiplication of virus-free good quality potato plantlets. Fifteen hormonal combinations were formulated and tested to determine a suitable hormonal combination and optimum concentrations for the in vitro production of a high number of plantlets for four farmer-preferred varieties in Niger (ATLAS, PAMELA, STEMSTER and YONA).
 Study Design: A completely randomized design (CRD) was employed to reveal the performance of four potato varieties, as affected by fourteen combinations of NAA and BAP with four replications. Results of the study were subjected to the analysis of variance, and significant differences among treatments were determined using GENSTAT12.01. Segregation between means was made according to the Student Newman-Keuls test, at 5% threshold.
 Place and Duration of Study: The experiment was carried out at the laboratory of Biotechnology and plant improvement of the institute of radio-isotopes in the University Abdou Moumouni of Niamey, in Niger (latitude 13°29' North and longitude 2°10' Est), for 28 days.
 Methodology: Uninodal stem explants of in vitro plantlets were cultured on full-strength Murashige and Skoog media (MS) supplemented with fourteen different combinations of α-Naphthalene Acetic Acid NAA (0; 0.25; 0.50; 1.0; 2.0 mg/l) and Benzyl Amino Purine BAP (0; 0.25; 0.50 mg/l), for 28 days.
 Results: Statistical analysis of the results showed that the variety and hormonal significantly influenced the height of the plant, the number of leaves and number of roots. Treatments with NAA alone without BAP stimulated rhizogenesis. From a NAA concentration of 0.25 to 0.5 mg/l there is a proliferation of the roots and from 1 mg/l there is an increase in roots length. PAMELA and ATLAS cultivars showed better root production.