scholarly journals In vitro culture of Dendrobium lineale Rolfe orchid for plant breeding and propagation

2021 ◽  
Vol 913 (1) ◽  
pp. 012066
Author(s):  
Nureni Dhuha Mustika ◽  
Endang Semiarti

Abstract D.lineale is included in the CITES appendix II due to D. lineale has great potential as a cut flower and the parent of hybrid orchids. In vitro culture can be used for mass propagation of orchids for commercial breeding as well as conservation. Basic culture medium with the addition of organic substances can increase the growth of plantlets. The objective of this research was to determine the best condition for in vitro culture of D.lineale orchids for producing a mass number of plantlets. Methods used include planting orchid seedlings in various variations of culture media, plantlet subcultures, observation of the anatomy of leaves and roots, as well as the measurement of chlorophyll content. This research uses ‘pisang raja’ as organic material in the culture medium. The highest chlorophyll content in the leaves was 1.58 mg/g, while the highest chlorophyll content in the roots was 0.34 mg/g. Both were found in medium NP+100 g/L bananas in light conditions (1317 lux). The anatomical structure of the leaves shows that the mesophyll is thicker in plants placed under lighting 1317 lux because it shows good effectiveness in photosynthesis. The anatomy of the orchid roots showed differences in diameter in each treatment because the addition of bananas to the medium stimulated rapid and abundant root formation, but the size was smaller. Based on these results, it can be concluded that the addition of 100 gr/L bananas as organic substances in in vitro medium culture increases the growth rate of D. lineale orchids, which will be beneficial for conservation efforts.

2021 ◽  
Vol 34 ◽  
pp. 03001
Author(s):  
Natalia Kovalenko ◽  
Nadezhda Polivara

This paper presents the results of the adjustment of 18 modifications of culture medium based on MS medium composition – Murashige and Skoog (1962) for in vitro rhizogenesis of 10 varieties of domestic plum. There were used 4 nutrient media with a full amount of macro- and microelements – MS1 with a different combination of phytohormones, 1 medium without hormones – MSc, 13 media – MS2 with half the amount of macronutrients, differing in hormonal composition. It was found that the maximum number of rooted microshoots from 21.2 to 52.2 % was on MS2–8 medium, from 11.2 to 43.1 % on MS2–5. The analysis showed that into the medium increases the percentage of microplants by 12-17 % comparing with the medium MS2–5 and MS2–7 without FA. The distinctiveness of nutrient media by the type of auxin (IAA, NAA, IBA) made it possible to clarify that IBA is the most optimal of the auxins, and the concentration of 1.7 mg/l is borderline for the growth reactions of plum varieties. It was revealed that in vitro root formation depends not only on the compositions of growth substances in the nutrient medium, but also on the genotype of the variety.


Lankesteriana ◽  
2015 ◽  
Vol 7 (1-2) ◽  
Author(s):  
Melania Muñoz ◽  
Víctor Jiménez

Slipper orchids belonging to the genus Phragmipedium (subfam. Cypripedioideae) are seriously threatened and therefore listed in Appendix I of CITES. Less research has been conducted in this genus than in others belonging to the same subfamily. In this work, we evaluated development of capsules (seed pods) from Phragmipedium humboldtii, P. longifolium and P. peaarcei from the time of pollination until opening. Moreover, seed viability was tested with the tetrazolium method in each of the capsules that were subsequently used to evaluate the effect of light and two culture media (Knudson C vs. Murashige and Skoog half concentrated) on in vitro asymbiotic germination and seedling growth. 100% of the pollinated flowers developed capsules, which differentiated in terms of length and diameter among species. While the length of the capsules remained constant during development, their diameter increased during the first 6-8 weeks and then stopped. Time required for maturity and opening of the capsules also varied among species (31 weeks in P. humboldtii, 16 weeks in P. longifolium and 9.5 weeks in P. percei). Seed viability differed among species as well, averaging 34.3% in P. humboldtii, 44.7% in P. longifolium and 82.3% in P. pearcei. Furthermore, seed viability of each capsule was used to adjust the germination rate measured in each case. While very few P. humboldtii viable seeds germinated under the conditions tested (2.9%), better results were observed in the other two species (close to 40% germination). No significant effect of light/darkness regime or of culture medium was observed on germination. However, better growth of the germinated embryos was observed with the Knudson C medium and darkness conditions. Further subculture of the gro- wing plantlets under light conditions induced development of roots and allowed successful acclimatization of seedlings in the greenhouse. 


Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.


2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.


2021 ◽  
Vol 16 (8) ◽  
pp. 118-120
Author(s):  
Renu Chandola ◽  
Anshulika Upadhyaya

Meizotropis pellita is a rare, endangered and threatened plant species found in Patwadanger forest. In nature this species grows from rootstock in precise and sensitive habitats. However, phenolic compounds provoke browning reaction which is responsible for lethal browning in plant tissue culture and hindering in vitro germination. The aim of this study was to minimize the effect of phenolic compound during seed germination. We evaluate different antibrowning treatments in Meizotropis pellita seeds. The seeds were pretreated in Polyvinylpyrrolidone and then treated seeds were cultured on MS media and sterilized seeds were directly inoculated on PVP containing MS media. These culture media incubate under dark and light conditions at different durations respectively. Our results showed that addition of antibrowning agent, PVP inhibit onset of browning. Dark and light condition also played a crucial role on onset of browning. Delayed and low intensity of browning were observed in case of seeds that were inoculated on MS media containing PVP and were kept in dark. The present research offers a positive in vitro seed germination protocol for Meizotropis pellita conservation.


Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.


Nativa ◽  
2018 ◽  
Vol 6 (1) ◽  
pp. 27
Author(s):  
Marcos Vinícius Marques Pinheiro ◽  
Ana Cristina Portugal Pinto De Carvalho ◽  
Fabrina Bolzan Martins

No intuito de elevar as taxas de sobrevivência durante a etapa de aclimatização e posterior plantio a campo, avaliou-se o enraizamento in vitro de bananeira cv. Pacovan, em diferentes concentrações de sais MS e de sacarose. Utilizou-se DIC, esquema fatorial (6x2x3), com seis meios de cultura [sendo três concentrações de nutrientes do meio MS (100%; 50% de macronutrientes; e 50% dos sais macro e micronutrientes), e duas concentrações de sacarose (1,5/3,0%)], dois fotoperíodos (12/16 h) e três tempos de cultivo (21, 28 ou 35 dias) e seis repetições/tratamento. Analisaram-se: altura da planta, número de folhas/planta, massas frescas e secas das partes aérea e radicular. Para altura da planta, massa fresca da parte aérea e radicular, o meio MS 50% dos sais + sacarose (1,5%) com fotoperíodo de 16 h e tempo de cultivo de 35 dias foi satisfatório. Para massa seca da parte aérea foi MS 50% de sais + sacarose (3%), e para massa seca da parte radicular, MS 100% + sacarose (3%) (em 12hs/28 dias e 16hs/21 dias). Para o alongamento/enraizamento in vitro da bananeira cv. Pacovan sugere-se MS 50% de sais (macro e micronutrientes), redução ou manutenção de sacarose (1,5 ou 3%) em 16h/35 dias de cultivo.Palavra-chave: Musa spp., propagação in vitro, sistema radicular. CHANGES IN CULTURE MEDIUM, PHOTOPERIOD AND TIME OF CULTIVATION AFFECT THE IN VITRO ELONGATION AND ROOTING OF BANANA CV. PACOVAN ABSTRACT:In order to achieve high rates of survival during the acclimatization and later planting in the field, was evaluated the in vitro of banana cv. Pacovan plants under different concentrations of sucrose and MS basal salt mixture. The experiment was assembled in a DIC, in 6x2x3, six different culture media [three different MS salt mixture concentrations (100%; 50% of macronutrients; and 50% of macro/micronutrients) and two sucrose concentrations (1.5/3%)], two photoperiods (12/16 hours) and three cultivation times (21, 28 or 35 days). Each treatment was composed by 6 replicates. Plant height, number of leaves/plant, fresh and dry weight of roots and shoots, were analyzed. Satisfactory results for plant height and shoot and root fresh biomass were observed in MS with macro/micronutrients (50%) + sucrose (3%), 16 hours/35 days. The highest values of shoot dry weight were observed in MS with macro/micronutrients (50%) + sucrose (3%); the highest root dry weight was achieved with MS 100% + sucrose (3%) (12hs/28 and 16hs/21 days). The suggested medium for the in vitro elongation and rooting stage of banana cv. Pacovan is the MS with 50% of salts (macro and micronutrients), reduction or maintenance of sucrose (1.5 or 3%) in 16h/35 days of cultivation.Keywords: Musa spp., in vitro propagation, root system. DOI:


Genetika ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 129-136
Author(s):  
Velichka Rodeva ◽  
Stanislava Grozeva ◽  
Velichka Todorova

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.


2021 ◽  
Vol 51 ◽  
Author(s):  
Kaliana Gottschalk de Freitas ◽  
José Carlos Sorgato ◽  
Jackeline Schultz Soares ◽  
Luan Marlon Ribeiro

ABSTRACT Some factors may influence the in vitro culture of native orchids, especially the culture medium, sealing system and light condition. This study aimed to evaluate the addition of banana pulp and peel in the formulation of culture media, sealing system and irradiance, in the in vitro growth of Cattleya nobilior Rchb. f. A completely randomized design was used in a factorial scheme with five culture media [Murashige & Skoog supplemented or not with banana pulp or peel (100 or 200g L-1)], two light conditions [3,000 K LED lamps (43 µmol m-2 s-1 or 86 µmol m-2 s-1)] and two sealing systems (natural and conventional ventilation), with six replicates of one vial each. The plant height, diameter of the largest pseudobulb, length of the largest root and of the largest leaf, fresh weight and number of leaves, roots and shoots were evaluated. The use of both the banana pulp and peel as additives in the culture medium formulation promoted the C. nobilior growth. The natural ventilation system increased the plant height, pseudobulb diameter and leaf length, while the conventional system promoted tillering.


2018 ◽  
Vol 12 (2) ◽  
pp. 117
Author(s):  
Cecília Moreira Serafim ◽  
Arlene Santisteban Campos ◽  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cecília Ribeiro de Castro ◽  
Ana Cristina Portugal Pinto de Carvalho

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.


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