Somatic embryogenesis in Citrus sinensis, C. reticulata AND C. nobilis x C. deliciosa

Most of the plant regeneration processes in citrus, through tissue culture, involve indirect somatic embryogenesis. The optimization of these processes is important for the development of in vitro plant improvement and micropropagation studies. Studies to evaluate the effect of different carbohydrates in somatic embryogenesis were conducted using calli from 'Ponkan' mandarin (Citrus reticulata, Blanco), 'Cravo' mandarin (C. reticulata), 'Itaboraí' sweet orange (C. sinensis L. Osbeck.), 'Valencia' sweet orange (C. sinensis) and 'Kinnow' mandarin (C. nobilis Loureiro x C. deliciosa Tenore). The culture medium used was MT supplemented with sucrose, galactose, glucose, maltose or lactose with the following concentrations of 18, 37, 75, 110, and 150 mM. The culture medium used for the maturation of somatic embryos had 0, 15, 29, 44, 58 and 73 mM of sucrose, in presence or absence of 0.5 g L-1 of activated charcoal. Seventy-three mM of sucrose with 0.1 mg L-1 of GA3 in the presence or absence 0.5 g L-1 of activated charcoal was also tested. Overall, the carbohydrates galactose or lactose induced a higher number of somatic embryos. Sucrose concentrations of 58 and 73 mM generated a higher number of plantlets from mature embryos of 'Ponkan' mandarin and 'Valencia' sweet orange.

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'Ponkan' mandarin (Citrus reticulata Blanco) immature fruits storage

Ciência e Agrotecnologia ◽

10.1590/s1413-70542006000500029 ◽

2006 ◽

Vol 30 (5) ◽

pp. 1017-1020

Author(s):

Moacir Pasqual ◽

Leonardo Ferreira Dutra ◽

Aparecida Gomes de Araujo ◽

Milene Alves de Figueiredo

Keyword(s):

Light Intensity ◽

Activated Charcoal ◽

Aerial Part ◽

Sweet Orange ◽

Citrus Reticulata ◽

Ms Medium ◽

Growth Room ◽

Polyethylene Bags ◽

Ponkan Mandarin

The aim of this work was to evaluate the effect of 'Ponkan' mandarin (C. reticulata) x 'Pêra' sweet orange (C. sinensis) immature fruits storage and sucrose concentrations on embryos in vitro culture. Fruits with 3 to 4 cm in diameter were harvested and placed inside black polyethylene bags with lateral openings and stored at 5±1ºC during 135 days. Every 15 days a sample was removed, its embryos were excised and individually inoculated in test tubes containing 15 mL of MS medium (Murashige & Skoog, 1962) with sucrose (0, 1.5, 3, 6, 12, 18 and 24 g L-1) and 0.3 mg L-1 GA3 and 1 g L-1 activated charcoal. Those treatments rested 48 hours in the dark and later in a growth room at 27 ± 1ºC with a 16-h photoperiod and 32 µmol m-2 s-1 light intensity. Immature fruits can be stored for posterior excision and embryos culture. Fruits with 120 days after the pollination can be stored for at most 135 days without damaging the embryos viability. It was observed a better development of the aerial part and root system of plantlets from 'Ponkan' mandarin x 'Pêra' sweet orange embryos in MS medium with 12-18 g L-1 sucrose.

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Somatic Embryogenesis and Plant Regeneration in Viola canescens Wall. Ex. Roxb.: An Endangered Himalayan Herb

Plants ◽

10.3390/plants10040761 ◽

2021 ◽

Vol 10 (4) ◽

pp. 761

Author(s):

Arun Kumar Khajuria ◽

Christophe Hano ◽

Narendra Singh Bisht

Keyword(s):

Somatic Embryogenesis ◽

Culture Medium ◽

Casein Hydrolysate ◽

Dichlorophenoxyacetic Acid ◽

Ms Medium ◽

Mature Embryos ◽

2,4 Dichlorophenoxyacetic Acid ◽

Root Axis ◽

Somatic Embryo Induction

Viola canescens Wall. ex. Roxb. is an important but threatened medicinal herb found at 1500–2400 m above mean sea level in the Himalayas. Overexploitation and habitat preference have put the plant under serious threat. Thus, the present study was undertaken to develop an efficient protocol for in vitro propagation via somatic embryogenesis. The results revealed that plant can be regenerated successfully through somatic embryogenesis using leaf derived calli. Regular subculturing of calli on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D)/indole-3-butyric acid (IBA)/kinetin (Kn) and varying combinations of 2,4-D+Kn induced somatic embryogenesis. The maximum average number of somatic embryos (SE) (19.15 ± 2.66) was induced on the medium with 0.15 + 0.05 mg L−1 of 2,4-D and Kn, respectively, and this medium was used as a control. To enhance somatic embryo induction, the control MS medium was supplemented with l-glutamine (200–400 mg L−1) and casein hydrolysate (1–4%). The maximum average number of SE (27.66 ± 2.67) and average mature SE (13.16 ± 3.48) were recorded on the medium having 2 % l-glutamine and 50 mg L−1 casein hydrolysate. The induced SE were asynchronous, so, to foster their maturation, the culture medium (free from growth regulators) was supplemented with abscisic acid (ABA) and silver nitrate (AgNO3). The maximum average number (35.96 ± 3.68) of mature SE was noticed on MS medium supplemented with 1.5 mg L−1 ABA. Mature embryos had two well-developed cotyledons and an elongated hypocotyl root axis. The development of SE into plantlets was significant for embryos matured on the medium with AgNO3 and ABA, with 86.67% and 83.33% conversion on the medium with 0.20 mg L−1 6-benzylaminopurine (BAP). The plantlets thus produced acclimatized in a growth chamber before being transferred to the field, which showed 89.89% survival. The plants were morphologically similar to the mother plant with successful flowering.

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Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Jurnal Hortikultura ◽

10.21082/jhort.v29n2.2019.p137-146 ◽

2020 ◽

Vol 29 (2) ◽

pp. 137

Author(s):

Fitri Rachmawati ◽

Dewi Permanik ◽

Ronald Bunga Mayang ◽

Budi Winarto

Keyword(s):

Somatic Embryogenesis ◽

Activated Charcoal ◽

Embryogenic Callus ◽

Culture System ◽

In Vitro Propagation ◽

Clonal Propagation ◽

Ms Medium ◽

Indirect Somatic Embryogenesis ◽

Half Strength

Protokol perbanyakan klonal yang efektif dan efisien sangat diperlukan untuk produksi benih berkualitas pada komersialisasi produk unggulan hasil pemuliaan. Penelitian bertujuan untuk mendapatkan protokol perbanyakan klonal Dendrobium ‘Balithi CF22-58’ melalui embriogenesis tidak langsung. Percobaan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Januari hingga Desember 2017. Penelitian ini menekankan pada penggunaan jenis eksplan, media, dan sistem kultur. Jenis eksplan yang diuji adalah tunas pucuk, tunas lateral, dan pangkal plantlet dengan tiga media inisiasi [½ Murashige and Skoog (MS) dikombinasikan dengan 1,5 mg/l thidiazuron (TDZ) dan 0,5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2,5 mg/l metathopolin (mT) dan 0,05 mg/l BAP (MI-2), dan 5 mg/l mT dan 0,05 mg/l BAP (MI-3)]; empat media proliferasi, yaitu ½ MS dengan kombinasi: MP-1 (0,75 mg/l TDZ + 0,25 mg/l BAP), MP-2 (1,5 mg/l TDZ + 0,5 mg/l BAP), MP-3 (2,5 mg/l mT + 0,05 mg/l BAP), dan MP-4 (5,0 mg/l+ 0,05 mg/l BAP); dua sistem kultur (padat dan cair); dan tiga media regenerasi MPP-1 (½ MS dengan vitamin penuh (1/2 MS-FV) + 2% charcoal); MPP-2 (½ MS-FV); dan MPP-3 (2 g/l Rosasol 18:18:18 TE). Percobaan disusun menggunakan rancangan acak kelompok faktorial dengan lima ulangan. Hasil penelitian menunjukkan bahwa inisiasi kalus embriogenik (KE) tertinggi, yaitu 38,3% dengan waktu inisiasi 16,8 hari dihasilkan dari eksplan pangkal plantlet pada medium MI-1. Medium MP-2 dan sistem kultur cair mampu mempertahankan proliferasi KE sampai 83,1% dengan rasio penggandaan 3,23 kali. Perkecambahan embrio terbaik sampai 86,9% embrio berkecambah dengan 18,2 kecambah per rumpun dalam waktu 21,3 hari, ditunjukkan pada medium MPP-1, sedangkan pembesaran plantlet terbaik mencapai tinggi plantlet sampai 5 cm, jumlah daun hingga 4,9 helai, dan jumlah akar 2,8, dengan 2,6 cm panjang akar dan 0,27 g bobot basah plantlet, diperoleh pada medium MPP-3. Perbanyakan anggrek dengan protokol ini diperkirakan dapat menghasilkan sekitar 3.000–4.000 plantlet/eksplan/tahun. Protokol hasil penelitian ini sangat potensial diaplikasikan pada perbanyakan klonal Dendrobium melalui kultur jaringan. KeywordsDendrobium; Embriogenesis somatik; Perbanyakan masal; Proliferasi; Sistem kultur AbstractThe effective and efficient clonal propagation protocol is significantly needed for producing qualified seedling for commercialization of superior breeding products. The objective of the study was to establish clonal propagation protocol for Dendrobium ‘Balithi CF22-58’ via indirect somatic embryogenesis. The study was conducted at the Tissue Culture Laboratory in Indonesian Ornamental Crops Research Institute from January to December 2017. The study emphasized to utilize explant source, culture media, and culture system. Explant types were shoot tip, lateral shoot, and basal part of plantlets; three initiation media [half strength Murashige and Skoog (MS) medium containing 1.5 mg/l thidiazuron (TDZ) and 0.5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2.5 mg/l metathopolin (mT) and 0.05 mg/l BAP (MI-2), and 5 mg/l mT and 0.05 mg/l BAP (MI-3)]; four proliferation media (half strength MS medium supplemented with: MP-1 (0.75 mg/l TDZ and 0.25 mg/l BAP), MP-2 (1.5 mg/l TDZ and 0.5 mg/l BAP), MP-3 (2.5 mg/l mT and 0,05 mg/l BAP), and MP-4 (5.0 mg/l and 0.05 mg/l BAP); two culture system were solid and liquid; and three regeneration media viz, MPP-1 (half strength MS medium with full vitamin and 2% activated charcoal); MPP-2 (MR-1 activated charcoal free), and MPP-3 (2 g/l Rosasol 18:18:18 TE). These experiments were arranged using a factorial randomized complete block design with five replications. Results of the study revealed that the highest initiation rate of embryogenic callus (EC) was up to 38.3% in 16.8 days after culture. The EC was regenerated from a basal part on MI-1 medium, MP-2 medium and liquid culture system were able to maintain proliferation of embryogenic callus up to 83.1% with 3.23 multiplication rate. The best embryo germination up to 86.9% with 18.2 germinated embryos per clump within 21.3 days was determined on MPP-1 medium. While the best plantlet performances with 5 cm height of plantlets, 4.9 number of leaves, 2.8 number of roots, 2.6 cm root length, and 0.27 g plantlet fresh weight was obtained MPP-3 medium. With this propagation protocol, 3,000 - 4,000 plantlets/explant/year can be produced. Results of the study have high potential to be applied for in vitro propagation of Dendrobiums.

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Increasing the somatic embryogenesis frequency of Panax vietnamensis Ha et Grushv. by disinfection of leaf explant using nano silver and the addition of nano silver in culture medium

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/18/3/12712 ◽

2020 ◽

Vol 18 (3) ◽

pp. 517-527

Author(s):

Do Manh Cuong ◽

Hoang Thanh Tung ◽

Hoang Dac Khai ◽

Vu Quoc Luan ◽

Vu Thi Hien ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Culture Medium ◽

Leaf Explants ◽

Dry Weight ◽

Contamination Rate ◽

Nano Silver ◽

Plantlet Formation

Somatic embryo is a developmental method for mass multiplication of valuable medicinal plants. In this study, leaf explants of Ngoc Linh ginseng were disinfected with nano silver at different concentrations and exposure times to eliminate infectious agents and induce embryogenic callus for the production of somatic embryos. The results show that the lowest contamination rate (20.00%) was observed when leaf explants were treated with 0.5 g/L nano silver for 15 minutes while the highest embryogenic callus induction rate (72.22%) and fresh weight (0.77 g) was determined at 0.2 g/L nano silver for 20 minutes. High frequency of somatic embryogenesis formation and germination were occurred on MS medium supplemented 1.0 mg/L 2,4-D; 0.5 mg/L NAA; 0.2 mg/L Kin and 1.6 mg/L nanosilver. After 8 weeks of culture, the number somatic embryos derived from nano silver treated-leaves was increased 2 times than non-treated explants. Addition of 1.0 mg/L NAA and 1.2 mg/L nano silver was showed the highest shoot and root length, root number, fresh and dry weight of plantlets. This research showed that pre-treatment and supplement of nano silver in culture medium is potentially useful for improving embryogenesis frequency, and plantlet formation of Ngoc Linh ginseng cultured in vitro.

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Genomic Methylated Cytosine Level during the Dedifferentiation and Cellular Competence in Coffea arabica Lines: Insights about the Different In Vitro Responses

Forests ◽

10.3390/f12111536 ◽

2021 ◽

Vol 12 (11) ◽

pp. 1536

Author(s):

João Paulo de Morais Oliveira ◽

Natália Arruda Sanglard ◽

Adésio Ferreira ◽

Wellington Ronildo Clarindo

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Coffea Arabica ◽

Cytosine Methylation ◽

Leaf Explants ◽

Embryogenic Calli ◽

Ploidy Stability ◽

Indirect Somatic Embryogenesis ◽

Cellular Competence

Coffea arabica genotypes present distinct responses in vitro, and somaclonal variation occurrence has been reported. Global cytosine methylation is one of the epigenetic mechanisms that influences the Coffea in vitro responses. We aimed to establish the indirect somatic embryogenesis in C. arabica ‘Catuaí Vermelho’, ‘Caturra’ and ‘Oeiras’, associate the distinct responses to the methylated cytosine genomic level, and check the ploidy stability. Leaf explants were cultured in callus induction and proliferation medium. The resulted calli were transferred to the regeneration medium, and the mature cotyledonary somatic embryos were transferred to the seedling medium. ‘Oeiras’ exhibited the highest number of responsive leaf explants, followed by ‘Caturra’ and ‘Catuaí Vermelho’. Global methylated cytosine level increased over time in the ‘Catuaí Vermelho’ and ‘Caturra’ friable calli, remaining constant in ‘Oeiras’. ‘Oeiras’ did not regenerate somatic embryos, while ‘Catuaí Vermelho’ exhibited the highest number. Somatic embryo regeneration was associated with the increase of the methylated cytosine level. However, the ‘Catuaí Vermelho’ embryogenic calli showed a lower methylated cytosine level than ‘Caturra’. Recovered plantlets exhibited the same 2C value and chromosome number to the explant donors. Therefore, cytosine hypermethylation occurred during C. arabica indirect somatic embryogenesis, influencing cell competence and somatic embryos regeneration.

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Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

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Influence of Abscisic Acid and Sucrose on Somatic Embryogenesis in CactusCopiapoa tenuissimaRitt. formamostruosa

The Scientific World JOURNAL ◽

10.1155/2013/513985 ◽

2013 ◽

Vol 2013 ◽

pp. 1-7 ◽

Cited By ~ 13

Author(s):

J. Lema-Rumińska ◽

K. Goncerzewicz ◽

M. Gabriel

Keyword(s):

Somatic Embryogenesis ◽

Abscisic Acid ◽

Somatic Embryos ◽

Sucrose Concentration ◽

Turgor Pressure ◽

Direct Somatic Embryogenesis ◽

Fresh Weight ◽

High Concentration ◽

Globular Stage ◽

Indirect Somatic Embryogenesis

Having produced the embryos of cactusCopiapoa tenuissimaRitt. formamonstruosaat the globular stage and callus, we investigated the effect of abscisic acid (ABA) in the following concentrations: 0, 0.1, 1, 10, and 100 μM on successive stages of direct (DSE) and indirect somatic embryogenesis (ISE). In the indirect somatic embryogenesis process we also investigated a combined effect of ABA (0, 0.1, 1 μM) and sucrose (1, 3, 5%). The results showed that a low concentration of ABA (0-1 μM) stimulates the elongation of embryos at the globular stage and the number of correct embryos in direct somatic embryogenesis, while a high ABA concentration (10–100 μM) results in growth inhibition and turgor pressure loss of somatic embryos. The indirect somatic embryogenesis study in this cactus suggests that lower ABA concentrations enhance the increase in calli fresh weight, while a high concentration of 10 μM ABA or more changes calli color and decreases its proliferation rate. However, in the case of indirect somatic embryogenesis, ABA had no effect on the number of somatic embryos and their maturation. Nevertheless, we found a positive effect of sucrose concentration for both the number of somatic embryos and the increase in calli fresh weight.

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Agrobacterium-mediated transformation of the wild orchid Cattleya maxima Lindl

Universitas Scientiarum ◽

10.11144/javeriana.sc23-1.amto ◽

2018 ◽

Vol 23 (1) ◽

pp. 89

Author(s):

Augusta Yadira Cueva-Agila ◽

Rino Cella

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Attractive Alternative ◽

Gene Encoding ◽

Anatomical Structures ◽

Large Numbers ◽

Hygromycin Resistance Gene ◽

Growing Conditions ◽

Somatic Embryogenesis Receptor Kinase

Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the propagative nature of PLBs and the easiness by which they can be generated, make these structures an attractive alternative to seed-mediated production for growing large numbers of plants. To increase somatic embryogenesis and optimize the procedure, PLBs of Cattleya maxima were transformed using the Agrobacterium tumefaciens method. The T-DNA carried a Hygromycin-resistance gene, a visible marker (GFP5-GUSA) and a rice gene encoding the Somatic Embryogenesis Receptor Kinase, deemed to be important for somatic embryogenesis. Treated PLBs generated somatic embryos developing Hygromycin-resistant plantlets. The insertion of T-DNA was confirmed by PCR, and GFP expression was observed using a fluorescent stereomicroscope. Transformed Cattleya maxima PLBs were more efficient in forming somatic embryos (60-80%) than untransformed controls (45-57%), and this contrast was maximized in hormone-free, Murashige and Skoog (MS) medium (80% of the transformed plants compared to 57% of the untransformed ones). This finding supports the notion that SERK plays an important role in Orchid embryogenesis.

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EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v17n1.2016.p9-16 ◽

2017 ◽

Vol 17 (1) ◽

pp. 9

Author(s):

Yosi Zendra Joni ◽

Riry Prihatini ◽

Darda Efendi ◽

Ika Roostika

Keyword(s):

Somatic Embryogenesis ◽

Plant Growth ◽

Embryogenic Callus ◽

Plant Growth Regulator ◽

Somatic Embryos ◽

Casein Hydrolysate ◽

Malt Extract ◽

Mass Propagation ◽

Embryogenic Callus Induction

Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.

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ACTIVATED CHARCOAL APPLICATION FOR THE MICROPROPAGATION OF Cattleya crispata (Thunb.) Van den Berg

Nativa ◽

10.31413/nativa.v9i4.12164 ◽

2021 ◽

Vol 9 (4) ◽

pp. 352-358

Author(s):

Denys Matheus Santana Costa Souza ◽

Sérgio Bruno Fernandes ◽

Letícia Vaz Molinari ◽

Maria Lopes Martins Avelar ◽

Gilvano Ebling Brondani

Keyword(s):

Endemic Species ◽

Activated Charcoal ◽

Culture Medium ◽

Genetic Conservation ◽

Campo Rupestre ◽

In Vitro Germination ◽

Germination Speed

Micropropagation is an alternative for the genetic conservation and propagation of endemic species from “Campo Rupestre Ferruginoso”, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, seedling number, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of seedlings of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots.

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