Meristem Culture Research Articles

The development of an in vitro floral culture transformation system for quinoa

AbstractBecause of its high-quality seed protein and ability to thrive in marginal habitats, Chenopodium quinoa has been identified as an important emerging grain crop for global food security. However, the lack of an efficient and robust transformation system has been a barrier for conducting the advanced genetic studies needed to better understand and improve the species for agronomic traits. Here we present a novel transformation system based on Agrobacterium-mediated transformation of in vitro floral culture. Quinoa floral cultures were established from inflorescences that naturally formed on plants grown in vitro. When placed on a cytokinin-containing medium, chopped inflorescences rapidly generated highly meristematic floral cultures, primarily composed of floral buds, flowers at various developmental stages, inflorescence shoots, and leafy structures. Transformation of these cultures with Agrobacterium carrying selectable and visual markers (NPTII and GUS) produced independent, stably transformed meristematic cultures resistant to paromomycin after an extended selection period (about 3 mo with sub-culture occurring every 15 d). Transformation frequency was about 20% and was calculated as the number of independent transformed events per initial number of floral culture explants used for transformation. In vitro flowers and inflorescences from putative transgenic events self-pollinated naturally and produced viable seeds that germinated without dormancy. We also demonstrated that flowering shoots could be successfully grafted onto wild rootstock to increase the number of seeds generated from T0 floral shoots. Molecular and phenotypic analysis of the progeny confirmed that the transgenes were stably integrated and inherited according to expected Mendelian ratios.

Meristem culture is a quite efficient method for the eradication of Potato Virus Y (PVY) from calibrachoa ‘Pampa Salmon INTA’

Meristem culture is a quite efficient method for the eradication of Potato Virus Y (PVY) from calibrachoa ‘Pampa Salmon INTA’

Determination of phosphinothricin and paromomycin selective concentrations for obtaining transgenic spelt plants

Aim. To determine the selective concentrations of phosphinothricin and paromomycin for the selection of transgenic plants of spelt wheat. Methods. Shoot apical meristem culture, mature embryo culture, Agrobacterium-mediated genetic transformation. Results. Isolation and cultivation of shoot apical meristems of seedlings from three spelt genotypes and mature embryos from three other genotypes were carried out. A high frequency (from 80 to 100 %) of callus induction from explants was observed. It was shown that the addition of 5 mg/l of phosphinothricin or 100 mg/l of paromomycin to the culture medium almost completely inhibited plant regeneration compared to the control. After Agrobacterium-mediated genetic transformation of calli with a vector containing the phosphinothricin-N-acetyltransferase gene, regeneration of spelt shoots for one genotype was observed on a selective medium with 5 mg/l phosphinothricin. Conclusions. The selective concentrations of herbicide and antibiotic for obtaining transgenic spelt wheat plants with the corresponding marker genes are 5 mg/l for phosphinothricin and 100 mg/l for paromomycin.

HSVd elimination from tomato using cryotherapy combined with exogenous HSVd-targeting ds-sRNA Application

The development of viroid elimination technology is essential in tomato (Solanum lycopersicum) production. Combining long-term cold therapy and meristem culture is commonly used; however, tomatoes cannot withstand cold. How the Hop stunt viroid (HSVd) affects the growth potential, activities of oxidoreductases and the contents of soluble matters and total chlorophyll was studied. HSVd-targeting double-stranded small RNAs (ds-sRNAs) were synthesized to match different regions of its genome (ds-sRNAs-H1∼H5), and their permeability in plants with HSVd inhibition effects was tested. The HSVd infected tomato shoots were exogenously treated with ds-sRNAs, and the meristems were thereafter taking from treated shoots and processed using leaf primordia-free meristem culture and cryotherapy. HSVd inhibited tomato stem elongation, leaf growth and root development, it elevated the oxidoreductase activity and disturbed the contents of soluble protein, soluble sugar and chlorophyll content. Exogenous ds-sRNAs can penetrate all types of cells to induce HSVd gene silencing in shoot terminals, causing the accumulation of miRNAs produced by HSVd and differed expression of gene silencing-related genes. The shoot terminal ex vivo treated with ds-sRNA which targeting the central conserved region of HSVd genome (H3) gained the highest inhibition of HSVd titer after 36 h of feeding. Leaf primordia-free meristem culture couldn't obtain viroid-free regenerants from ds-RNA-H3 treated meristems, whereas in cryotherapy, meristems soaked in PVS2 for 15 and 20 min resulted viroid elimination of 6.7 % and 10.0 %, 5 and 10 min of PVS2 caused 0 % HSVd elimination. Growth and physiological state of HSVd-eliminated plants recovered to those of healthy controls. The results provide theoretical and technical basis for the sustainable development of the tomato pathogen-free plant industry.

Optimizing sweet potato production: insights into the interplay of plant sanitation, virus influence, and cooking techniques for enhanced crop quality and food security.

This study investigates the impact of sweet potato plant sanitation on the yield and external and internal quality root storage exploring the nutritional content affected by various cooking methods (raw, boiled, and oven-cooked). The presence of viruses, and concretely of the sweet potato leaf curl virus (SPLCV), in sweet potato propagation material is shown to significantly reduce yield and modify storage root quality. Notably, the research reveals a substantial improvement in crop yield and external quality, reinforcing the efficacy of plant sanitation methods, specifically apical meristem culture, in preserving the overall productivity of sweet potato crops. Furthermore, the investigation identifies a noteworthy decrease in starch content, suggesting a dynamic interaction between plant sanitation and starch metabolism in response to viral diseases. The study also delves into the alteration of mineral absorption patterns, shedding light on how plant sanitation influences the uptake of essential minerals in sweet potato storage roots. While the health status of the plants only slightly affected magnesium (Mg) and manganese (Mn) accumulation, indicating a potential resilience of mineral balance under virus-infected conditions. Moreover, the research identifies significant modifications in antioxidant levels, emphasizing the role of plant sanitation in enhancing the nutritional quality of sweet potatoes. Heat-treated storage roots, subjected to various cooking methods such as boiling and oven-cooking, exhibit notable differences in internal quality parameters. These differences include increased concentrations of total soluble solids (SS) and heightened levels of antioxidant compounds, particularly phenolic and flavonoid compounds. The observed increase in antioxidant capacity underscores the potential health-promoting benefits associated with plant sanitation practices. Overall, the study underscores the critical importance of plant sanitation in enhancing sweet potato production sustainability, contributing to food security, and supporting local agricultural economies. The results emphasize the need for further research to optimize plant sanitation methods and promote their widespread adoption globally, providing valuable insights into the complex relationships in food quality.

Evaluation of Virus-Free Chrysanthemum 'Hangju' Productivity and Response to Virus Reinfection in the Field: Molecular Insights into Virus-Host Interactions.

The shoot apical meristem culture has been used widely to produce virus-free plantlets which have the advantages of strong disease resistance, high yield, and prosperous growth potential. However, this virus-free plant will be naturally reinfected in the field. The physiological and metabolic responses in the reinfected plant are still unknown. The flower of chrysanthemum 'Hangju' is a traditional medicine which is unique to China. In this study, we found that the virus-free 'Hangju' (VFH) was reinfected with chrysanthemum virus B/R in the field. However, the reinfected VFH (RVFH) exhibited an increased yield and medicinal components compared with virus-infected 'Hangju' (VIH). Comparative analysis of transcriptomes was performed to explore the molecular response mechanisms of the RVFH to CVB infection. A total of 6223 differentially expressed genes (DEGs) were identified in the RVFH vs. the VIH. KEGG enrichment and physiological analyses indicated that treatment with the virus-free technology significantly mitigated the plants' lipid and galactose metabolic stress responses in the RVFH. Furthermore, GO enrichment showed that plant viral diseases affected salicylic acid (SA)-related processes in the RVFH. Specifically, we found that phenylalanine ammonia-lyase (PAL) genes played a major role in defense-related SA biosynthesis in 'Hangju'. These findings provided new insights into the molecular mechanisms underlying plant virus-host interactions and have implications for developing strategies to improve plant resistance against viruses.

Elimination of yam mosaic virus from yam using an optimized combination of meristem culture and thermotherapy

Elimination of yam mosaic virus from yam using an optimized combination of meristem culture and thermotherapy

IN VITRO PRODUCTION OF VIRUS-FREE CARNATION (DIANTHUS CARIOPHYLLUS L.) PLANTING MATERIAL

The methods of culture of apical meristems and direct and indirect morphogenesis in vitro were used for production of virus-free planting material of carnation. A scheme for obtaining aseptic material has been developed, which consists of stepwise treatment of explants: Thimerosal - 2 min, 70% ethyl alcohol - 0.5 min and 0.08% AgNO 3 - 1 min, which reduces the level of contamination by fungal infection. Expounded the results of studies of callusogenesis and direct and indirect morphogenesis in the culture of in vitro explants of Dutch carnation, their dependence on the content of growth regulators in the nutrient medium. It was established that there were almost no significant differences in the course of callusogenesis processes within carnation varieties . At the same time, the frequency of callusogenesis was 100%. Under the conditions of indirect morphogenesis realization, it is necessary to take into account the age of callus tissues. The growth and intensive shoot formation of carnations was noted on the Murashige-Skoog nutrient medium supplemented with BAP at a concentration of 0.5 mg/l. The best medium for rooting was the MS medium with half the concentration of macro- and microsalts with the addition of 0.5 mg/l of NAA, which is recommended by us for rooting regenerating carnation plants of various varieties. Peat : perlite in a 1:1 ratio was used as a substrate for the adaptation of regenerating plants . Survival of carnation plants to conditions in vivo for the variety "Raffino Linde" was 90%, while for the variety "Tiya" - 83%, respectively.

Phytofabrication of silver nanoparticles using callus extracts of natural tetraploid Trifolium pratense L. and its bioactivities

One of the main subjects of plant biotechnology is plant tissue culture and in recent years is considered a possible approach model for green and eco-friendly biosynthesis of nanoparticles. This study aimed to present calli produced from the natural tetraploid Trifolium pratense L. containing high amounts of phenolic compounds and glycosidic bioactive macromolecules and the biosynthesis of silver nanoparticles from calli. Combinatorial optimization of silver nanoparticles was achieved for the first time in this study, thanks to the stabilizing and reducing properties of hypocotyl, apical meristem, and epicotyl derived callus extracts of the natural tetraploid T. pratense L. biosynthesized nanoparticles from three different callus extracts. Callus extracts were used to create different experiments with AgNO3 at various concentrations (0.16, 0.5, 0.84, 1.18, 1.52 and 1.96 mg L-1), different temperatures (40, 50, 60, 70, 80, 90, 100°C), and different pH levels (5, 7, 10) to carry out the biosynthesis of AgNPs. Biologically synthesized AgNPs were easily monitored by color change in ultraviolet and UV-Vis spectroscopy proved to be a fast and simple method. Also, TEM, XRD, and FTIR analyses were done to characterize and confirm the formation of crystalline nanoparticles. It was determined that antibacterial activity inhibition was achieved by using the Agar-well diffusion method for antibacterial activity measurements on Gram-positive Staphylococcus aureus ATCC 25923 and Gram-negative Escherichia coli CECT 4972 bacteria. Biosynthesized AgNPs were observed in the wavelength range of 400-500 nm in the UV-VIS spectrum. TEM analysis demonstrated the size and shape of biosynthesized silver nanoparticles under different conditions. It was observed that the smaller silver nanoparticles were spherical and the larger silver nanoparticles were triangular, elliptical, and spherical shape. The XRD analysis proved the presence of Ag0 in nanoparticles and showed crystal structure for silver nanoparticles. By FTIR analysis, O-H hydroxyl groups of functional groups on the AgNP surface, H-linked OH stretching, C-H stretching, -CH stretching of -CH2 and -CH3 functional groups, C-N and carboxylate, aliphatic phosphate and primary amine stretching were expressed. Biosynthesized silver nanoparticles showed antibacterial activity against Gram-positive S. aureus ATCC 25923 bacteria, AgNP hypocotyl (1.7mm), AgNP-epicotyl (1.1mm) against Gram-negative E. coli CECT 4972 bacteria. Among the hypocotyl, apical meristem, and epicotyl callus cultures, the highest antioxidant activity was observed in the AgNPs obtained from hypocotyl-concentration experiments, with a DPPH radical activity of 52% and an ABTS radical activity of 68%. In conclusion, these findings underscore the potential of biotechnological strategies in green nanotechnology, which can be offered for developing metal nanoparticles with potential biomedicine and biotechnology applications.

Micropropagation of Duboisia Species via Shoot Tip Meristem

Duboisia is an Australian native, commercially valuable for tropane alkaloid extraction. Clonal propagation of elite selections is essential to establish highly productive plantations. The current propagation system using stem cuttings is proven to be inefficient, prompting the industry to seek a more efficient and effective propagation tool. Tissue culture is a cost-effective alternative for mass propagation of true-to-type plants, particularly ideal for propagating elite Duboisia selections. In this context, attempts were made to develop a commercially viable high throughput micropropagation system for three Duboisia species: Duboisia myoporoides, Duboisia leichhradtii and Duboisia hopwoodii. Various nutrient media, hormone combinations and incubating conditions were tested to optimise each stage of the micropropagation pipeline. The findings revealed that the tissue culture media composition and hormone requirements are species-specific. With the optimised conditions, an efficient tissue culture system was developed, achieving successful meristem induction and multiplication. Species-specific rooting protocol optimisation resulted in 100% rooting for D. myoporoides and D. leichhardtii, and 70% rooting for D. hopwoodii. Furthermore, an optimised acclimatisation protocol supported 100% survival of D. myoporoides and D. leichhardtii and 80% of D. hopwoodii plantlets. This study, for the first time, demonstrated the capacity of successful meristem culture of three Duboisia species, establishing the foundation for high throughput micropropagation of Duboisia species.

Biotechnological Strategies Adopted for Sugarcane Disease Management in Tucumán, Argentina

Sugarcane diseases can be controlled by an integrated management approach where biotechnological tools can successfully contribute. The Obispo Colombres Agroindustrial Experimental Station (EEAOC) in Tucumán (Argentina’s main sugarcane producer) has successfully implemented multiple strategies that greatly enhance the productivity of sugarcane fields. The local breeding program develops resistant varieties by applying molecular markers to reveal the presence of Bru1 gene for brown rust resistance throughout the EEAOC germplasm collection. In addition, SNP alleles linked to novel sources of resistance were identified following a selective genotyping strategy. Another strategy is the implementation of a seed cane sanitation project using hydrothermal therapy, an in vitro culture technique, molecular diagnosis of diseases, and bionanoparticles. As a result, the incidence of systemic diseases has significantly decreased in the production fields. More recently, the use of biological products has shown to be effective for disease control in EEAOC varieties. In summary, several biotechnological strategies including molecular markers associated with resistant sources, in vitro culture of apical meristems, molecular diagnostic techniques, and the use of bioproducts are being successfully used for the sustainable management of sugarcane diseases in Tucumán, Argentina.

In vitro propagation and microtuberization of potato (Solanum tuberosum L.) Spunta variety in Lebanon

One of the factors that causes low productivity of potatoes in Lebanon is the limited availability of certified seeds. The aim of this study was to establish a rapid protocol for in vitro propagation and microtuberization of potato (Solanum tuberosum L.) of Spunta variety. Meristems culture associated to thermotherapy (one month/37°C) constituted the first step. The highest percentage of reactive meristem (92%) was observed on MS medium devoid of growth regulators while MS medium containing Kin 0.4 mg.l-1, GA3 0.5 mg.l-1 and IBA 0.5 mg.l-1 yielded the highest average number of shootlets (7.8) in the seventh subculture. The lowest number of days obtained for microtuber formation was 10 and the highest average number of microtuber (1.49) was obtained with shootlets incubated under C2 culture conditions (16-h day/8-h night for initial 7 days at 25±2°C; for remaining period: continuous dark at 17±2°C). Contrary the highest microtubers average length (10.75 mm), average width (7.41 mm) and average weight (646.26 mg) were produced under C1 culture conditions (16-h day/8-h night at 25±2°C). Medium supplemented with 5 mg.l-1 BAP and 6% sucrose presented the highest average number of microtubers of 2.36 and 1.94 respectively. Type and concentration of cytokines and sucrose concentration did not have significant effect on the average length, width and weight of microtubers produced.

Pome fruit-virus interactions using combined therapies and meristem culture

Pome fruit-virus interactions using combined therapies and meristem culture

The Effect of Light Quality and Intensity on <em>in vitro</em> Potato Cultures

Purpose: The study investigates the effect of light quality and intensity factors on in vitro potato meristem cultures and multiplication for optimization of pre-basic seed potato productions of some potato varieties.Research Method: In vitro experiments were conducted to study the response of meristems and nodal cuttings of five potato varieties, i.e., Cara, Hermes, Lady Rosetta, Santana, and Spunta, to four LED light qualities (blue, red, red+blue, or white) for five potato varities. Also, the response of nodal cuttings was examined under three light intensities (50, 75 and 100 μmol m-2 s-1).Findings: Significant differences were obtained between the four tested light qualities. Red LED gave the best meristem survival rates of the Cara, Hermes, Lady Rosetta, and Spunta potato varieties. In the multiplication phase, a significantly (p ≤ 0.05) higher plantlet length was obtained from nodal cuttings under red light quality. Also, white and red light produced vigorous plantlets, expressed as higher significant dry weight (82.2 and 80.4 mg/plantlet, respectively). Increasing light intensity from 50 to 75 and 100 μmol m-2 s-1 resulted in increases in leaf number, stem diameter, root length, leaf area, chlorophyll content, fresh weight, and dry weight.Originality/value: White LED light quality enhanced in vitro initiation of potato meristems. Furthermore, Light intensity of 75 μmol m-2 s-1 gave better performance of potato plantlets in vitro.

IMPROVEMENT OF APPROACHES TO ELIMINATION OF VIRUSES IN POTATOES USING IN VITRO CULTURE

Background. Potatoes are considered to be easily susceptible to infections, including viral diseases, which is one of the reasons for the decrease in yield. To date, the only ways to obtain and reproduce healthy potato material are technologies based on meristem-tissue cultures.
 Purpose. The aim of this study was to improve approaches to the release of potato from viral infection using in vitro culture.
 Materials and methods. The objects were tubers, microplants and microtubers of six cultivars of potato affected by PVS, PVA, PVM, PVX and PVY viruses in various combinations. Viruses were eliminated from plant material by thermotherapy at a temperature of 38.0±0.5°C. Depending on the cultivar and object, the exposure was 10-25 days.
 Results. The study showed that the use of potato microtubers as an object for thermotherapy makes it possible to release plant material from viruses in 25.0-50.0% of cases, depending on the pathogen. The advantage of this virus eradication scheme is, firstly, in the increase in the regenerative capacity of explants up to 32.9% due to the exclusion of the sterilization stage after thermotherapy and the absence of apical meristem culture as an additional method of sanitation. Secondly, the efficiency of the release of plant material from viruses increases due to a longer exposure to high temperature up to 25 days.
 Conclusion. The use of this approach made it possible to eliminate not only individual viruses, but also their complex, for example, PVA + PVX, PVM + PVX, PVX + PVS, PVM + PVX + PVS.

Effect of Viusid Agro® on the Growth of Banana (Musa Spp.) Seedlings Under Nursery Conditions

The commercial production of plantains and bananas (Musa spp.) involves several aspects to overcome, including those ones related to abiotic and biotic factors. Catalysis, S. L. Agroveterinary Division has developed the product VIUSID agro® as a plant growth promoter in agriculture. Research centers from several countries have shown field studies that reveal the product efficacy. Studies conducted on this topic support the importance of the use of this product in crops, such as cassava, papaya, pumpkin, cucumber, bean, chickpea, garlic, among others at the Research Institute of Tropical Roots and Tuber Crops (INIVIT). This research is carried out with the aim of determining the effect of VIUSID agro® on the growth of banana seedlings. The experimental design was conducted in February 2020, using banana Musa AAA seedlings, Cavendish subgroup, 'Gran enano' cultivar of six months of age from meristem cultures, transplanted to polyethylene bags with a mixture of red soil and organic matter (70:30%), located at the Center for Accelerated Seed Reproduction (CRAS). VIUSID agro® (0,2 mL L-1) was applied as foliar application during the first, third and fifth weeks after transplanting. The treated seedlings showed significant statistical differences in terms of: basal diameter (mm), height (cm), number of leaves, foliar area (cm2), radial length of emerged roots directly from the corm (cm), biomass of the foliar and belowground area with respect to the control treatment. VIUSID agro® is an excellent biostimulant, which allows the farmer to take to his farms more vigorous and better adapted seedlings to adverse conditions.

Advancements in Lily Viruses Management: Challenges and Solutions in Elimination and Detection

Lilies are important crops that are commonly used as cut flowers (Lilium spp.) and edible bulb crops (Lilium davidii var. unicolor). However, virus infections can significantly impact the quantity and quality of lily production. Various methods have been developed to eliminate viruses in lilies, including in vitro culture and virus detection techniques. Meristem culture is the most effective method, which can be combined with other techniques such as thermotherapy and chemotherapy. Nonetheless, virus elimination is affected by several factors, including cultivar, explants used, virus type, and duration of treatments. Efficient diagnostic methods, such as serological and molecular techniques, have been developed to detect viral infections in lilies, including enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR). However, cross-contamination and multiple-virus contamination can lead to unreliable results, and more sophisticated protocols and systems have been developed to address these issues. The objective of this review is to provide a comprehensive overview of the development of lily virus eradication, detection strategies, challenges, and solutions associated with these procedures, and how more sophisticated approaches such as multiplex RT-PCR, indirect ELISA (ID-ELISA), immunocapture RT-PCR (IC-RT-PCR), and immunochromatographic test strips (ICSs) can alleviate some of these setbacks.

Effects of thermotherapy and meristem culture techniques on macro and micronutrients content in elephant grass cultivars

Elephant grass is a tropical forage crop highly used in dairy cattle production, in Brazil. It has been getting special attention, because of its bioenergy potential, medicinal properties, and bioremediation profile. The aim of the present study is to investigate the effects of thermotherapy based on clonal cleaning methods and meristem culture on the mineral content of elephant grass (Cenchrus purpureus (Schumach.) Morrone). Cultivars “Mineiro”, “Taiwan A-147” and “Pioneiro” were subjected to the following methods: thermotherapy (T) combined to meristem culture (MC), meristem culture and mature stems (control). The experiment assessed the mineral contents of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg) and sulfur (S) at three cuttings, which were performed every 60 days, for 180 days. There was lack of effects from these methods on the mineral content of approximately 66% of the carried out assessments, standing out unanimity for all cultivars, methods, and most of the cuts. T+MC was the only method showing positive effect on P and Ca content, in all cuts, in the cultivars Taiwan A-147 and Pioneiro, respectively. There was clear negative effect of cleaning methods on P and Ca content, in all cuts, for cultivar Mineiro, and on Mg, for cultivar Taiwan A-147. These results, along with the positive effects observed in vegetative and nutritional parameters shown in other articles published in this Journal, show that the clonal cleaning methods are strongly recommended for cultivars with more than 15 years of ripe stem propagation.

Micropropagation of strawberry (Fragaria ananassa “pajaro”)by meristem culture method

Experiments were carried out to examine the effects of different combinations of plant growth regulators in vitro micropropagation of strawberry plants. Specimens were sterilised by a different concentration of sodium hypochlorite - NaOCl (1, 2, and 3%) for 20 mins. A meristem was separated and cultured on Murashige and Skoog (MS) containing 0.3 mg/l BAP for 8 weeks. Initial shoots were transferred to MS with various concentrations of BAP (0, 0.1, 0.3, and 0.5 mg/l) and kinetin (0, 0.1, 0.2, and 0.3 mg/l) in either single or combination. After 6 weeks of culture, the regenerated shoots were transferred on 1/2 MS with different concentrations of NAA (0, 0.1, 0.3, 0.5, 0.7, and 1.0 mg/l) and BAP (0, 0.1, 0.3, and 0.5 mg/l) in either single or combination. The results showed that the lowest percentage of contamination (10%) and the highest percentage of regeneration (80%) response were achieved on MS media by sterilisation with 3% NaOCl for 20 mins. The highest shoot proliferation per explant (16.2 shoots/explant) was reached on MS with 0.3 mg/l BAP plus 0.1 mg/l kinetin. 1/2 MS media with 0.1 mg/l NAA plus 0.1 mg/l BAP induced 14.4 roots per shoot and produced the longest roots (9.1 cm). However, the higher concentration of NAA (0.3-1.0 mg/l) or in combination with BAP resulted in callus formation. The plantlets, thus developed, were hardened and successfully established in soil containing organic fertiliser: Trichoderma: coconut peat (1:1:1 v/v/v).

Shoot and Callus Formation by Type, Gelling agent, pH and Sucrose Concentration in Medium in Apical Meristem Culture of Rosa multiflora ‘Natal Briar’

Shoot and Callus Formation by Type, Gelling agent, pH and Sucrose Concentration in Medium in Apical Meristem Culture of Rosa multiflora ‘Natal Briar’