Nutritional requirements for germination and in vitro development of three Orchidaceceae species in the southern Brazilian Amazon

Tissue culture is an alternative form of producing healthy, vigorous and regular plants on a large scale. The purpose of this study was to evaluate the most efficient culture medium for in vitro plantlet germination and development of three Orchidaceae species. Seeds disinfested of three species were dispersed in distilled water and dripped into basic Murashige and Skoog (MS) medium. The experimental design was completely randomized in a factorial 3 x 4 (three species x four culture media), with 5 replications. Four treatments were established: (1) full-strength MS medium, with the full nutrient concentration (MSØ), (2) full-strength MS medium plus 0.3% activated charcoal (MSØ ACh), (3) half- strength MS medium (½ MS) and (4) half- strength MS medium with 0.3 % activated charcoal (½ MS ACh). Germination was evaluated after 15, 20, 25, 30, and 60 days. The shoot height, leaf number and length, root number and length of plantlets of the three studied species were assessed. In A. variegata, 73% germinated after 60 days in ½ MS ACh medium. In the same period, 100% of E. viparum and S. gloriosa seeds germinated in MSØ ACh medium. The plant height, leaf number and length, root number and length were significantly higher for the species A. variegata and E. viviparum in MSØ ACh medium. The culture media ½ MS and MSØ with addition of activated charcoal favored in vitro germination for the three orchid species of this study.

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Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽

10.21273/hortsci.44.1.106 ◽

2009 ◽

Vol 44 (1) ◽

pp. 106-112 ◽

Cited By ~ 15

Author(s):

Alice Noemí Aranda-Peres ◽

Lázaro Eustáquio Pereira Peres ◽

Edson Namita Higashi ◽

Adriana Pinheiro Martinelli

Keyword(s):

New Media ◽

Mineral Composition ◽

Culture Media ◽

Dry Weight ◽

Defined Medium ◽

Ms Medium ◽

Media Formulation ◽

Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

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In vitro culture and acclimatization of Cattleya xanthina (Orchidaceae), an endangered orchid of the Brazilian Atlantic Rainforest

Rodriguésia ◽

10.1590/2175-7860201970014 ◽

2019 ◽

Vol 70 ◽

Cited By ~ 1

Author(s):

Monique Cristine R. Juras ◽

Jackeline Jorge ◽

Rosete Pescador ◽

Wagner de Melo Ferreira ◽

Vivian Tamaki ◽

...

Keyword(s):

Photosynthetic Pigments ◽

Total Protein ◽

Culture Media ◽

Atlantic Rainforest ◽

Ms Medium ◽

Ammonium Ions ◽

Biometric Parameters ◽

Half Strength ◽

Brazilian Atlantic Rainforest

Abstract Cattleya xanthina is a Neotropical orchid endemic to the Brazilian Atlantic Rainforest, at high risk of extinction. In this paper, we investigated the effects of different culture media on C. xanthina as well as on their endogenous nitrogen status. Culture media studied: Knudson C (KC), Vacin and Went (VW), and Murashige and Skoog (MS), the latter used at two different concentration (full and half-strength; MS/2). After 180 days, plants were transferred to MS medium with different NAA and BA concentrations. In each treatment, biometric parameters were measured and the endogenous levels of photosynthetic pigments, total protein, nitrate and ammonium ions were quantified. Plants grown on KC medium had the lowest concentration of nitrogen but exhibited the greatest shoot development, production of photosynthetic pigments and total protein. Results of growth regulators showed that the highest concentration of auxin stimulated root development and the production of photosynthetic pigments, and that a higher concentration of cytokinin promoted protein synthesis and the development of shoots. Most successful acclimatization was obtained when a mixture of Sphagnum and Pinus bark was used as the substrate.

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In vitro and ex vitro rooting of Spiraea betulifolia subsp. aemiliana (Rosaceae), an ornamental shrub

BIO Web of Conferences ◽

10.1051/bioconf/20202400058 ◽

2020 ◽

Vol 24 ◽

pp. 00058

Author(s):

Dinara Muraseva ◽

Vera Kostikova

Keyword(s):

Ms Medium ◽

Pulse Treatment ◽

Root Number ◽

Ex Vitro ◽

Effective Technique ◽

Nutrient Media ◽

Half Strength ◽

Regenerated Plantlets ◽

Ornamental Shrub

Two methods of rhizogenesis – in vitro and ex vitro of Spiraea betulifolia subsp. aemiliana (C.K. Schneid.) H. Hara microshoots have been compared. Pulse treatment of microshoots with aqueous solutions of 4% “Heteroauxin” or 2% succinic acid (ex vitro rooting) did not effective -rooting frequency ranged from 3 to 19%. It was established that the in vitro rooting on nutrient media supplemented with auxins was a more effective technique, providing a high percentage of rooted microshoots. The use of half- strength MS medium supplemented with 0.1 μM indolyl-3-butyric acid (rooting frequency 8 8%, root number 3.5 ± 0.3 per plantlet) was found to be the most efective for in vitro rooting. The in vitro rooted regenerated plantlets were successfully acclimatized with 55% of survival rate.

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Effect of Seed Orientation and Medium Strength on In vitro Germination of Pterocarpus marsupium Roxb.

Notulae Scientia Biologicae ◽

10.15835/nsb549124 ◽

2013 ◽

Vol 5 (4) ◽

pp. 476-479 ◽

Cited By ~ 3

Author(s):

Yogeshwar MISHRA ◽

Rimi RAWAT ◽

Brajesh NEMA ◽

Fatima SHIRIN

Keyword(s):

Shoot Length ◽

Ms Medium ◽

Root Number ◽

Pterocarpus Marsupium ◽

Shoot Number ◽

Medium Strength ◽

Half Strength ◽

Varied Response ◽

Strength Medium

A method was developed for optimization of In vitro germination of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. The seeds inoculated in different orientation on different strengths of MS medium without any hormones showed varied response to the seed positions. The seeds inoculated on half strength medium in horizontal position recorded to produce a maximum germination (78.23%), shoot number (0.86) and root number (7.99). However, a maximum of shoot length of 3.67 cm was recorded in the quarter strength medium in the seeds inoculated in vertical down position, which was significantly higher than other media strength and positions. Our results indicate that the seed orientation including medium strength have tremendous effect on germination and seeds inoculated horizontally on half strength MS medium can be utilized to enhance In vitro seed germination of Pterocarpus marsupium.

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Study on in vitro propagation of Sophora tonkinensis Gagnep through stem node culture

Ministry of Science and Technology, Vietnam ◽

10.31276/vjst.63(12).59-63 ◽

2021 ◽

Vol 63 (12) ◽

pp. 59-63

Author(s):

Thi Lai Nguyen ◽

◽

Thi Binh Nguyen ◽

Anh Duc Pham ◽

Huong Son Pham ◽

...

Keyword(s):

Activated Charcoal ◽

Coconut Water ◽

High Survival ◽

Ms Medium ◽

Shoot Height ◽

Sophora Tonkinensis ◽

Carrot Puree ◽

Node Culture ◽

Stem Segments

In the present study, authors propagated Sophora tonkinensisGagnep plants using stem nodal culture. The results indicated that on MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 0.75 mg/l TDZ shoots proliferated from stem segments have the best count and height of shoots. The most appropriate medium for multiplication of shoots was the MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 0.75 mg/l TDZ, 0.5 mg/l IBA, 2.0 g/l peptone, 30 g/l carrot puree, pH 5.5 with the results of 20.60 shoots/explant, shoot height of 3.75 cm and 4.6 leaves/shoot after 8 weeks of culture. Root formation of shoots carried out on the MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 1.0 mg/l αNAA gave the best result. In the nursery, a mixture of humus + coconut fiber powder (70:30 ratio) was regarded as the best substrate due to the high survival rate of plantlets (92%) and healthy plantlets (10.30 cm high with 7.2 leaves and 4.3 new roots/a plantlet) at 10 weeks after planting

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Clonal Micropropagation of representatives of the genus Dactylorhiza Neck. ex Nevski

АгроЭкоИнфо ◽

10.51419/20214417 ◽

2021 ◽

Vol 4 (46) ◽

pp. 17-17

Author(s):

Alexander Saakian ◽

◽

Keyword(s):

Survival Rate ◽

Growth Regulators ◽

In Vitro Propagation ◽

Culture Medium ◽

Culture Media ◽

Ms Medium ◽

Organic Additives ◽

Clonal Micropropagation ◽

Half Strength

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES

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Direct Organogenesis from Rhizome Explants in Marsilea quadrifolia L.: A Threatened Fern Species

Advances in Biology ◽

10.1155/2015/639678 ◽

2015 ◽

Vol 2015 ◽

pp. 1-6 ◽

Cited By ~ 2

Author(s):

Mahipal S. Shekhawat ◽

M. Manokari

Keyword(s):

Multiple Shoots ◽

Direct Organogenesis ◽

Root Induction ◽

Ms Medium ◽

Full Strength ◽

Marsilea Quadrifolia ◽

Fern Species ◽

Half Strength

An efficient micropropagation protocol has been developed for Marsilea quadrifolia L. through direct organogenesis. The mature rhizomes were used as explants and successfully sterilized using 0.1% HgCl2 for the establishment of cultures. The multiple shoots were differentiated from the explants on Murashige and Skoog (MS) medium augmented with 6-benzylaminopurin (BAP). Full strength MS medium was reported to be effective for the induction of sporophytes from the rhizomes after four weeks of inoculation. Maximum response (96%) with average of 6.2 shoots (2.72 cm length) was achieved on full strength of MS medium augmented with 0.5 mg/L BAP in culture initiation experiments. The cultures were further proliferated in clusters (79.0±0.37 shoots per explant) with stunted growth on half strength MS medium supplemented with 0.25 mg/L BAP after four weeks. These stunted shoots were elongated (5.30 cm long) on half MS medium devoid of growth hormones. Root induction and proliferation (3.0–4.0 cm long) were observed after 4th subculture of sporophytes on hormone-free half strength MS medium. The rooted plantlets were hardened in the fern house for 4-5 weeks and transferred to the field with 92% survival rate. There were no observable differences in between in vivo grown and in vitro propagated plantlets in the field.

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In vitro propagation and growth of ex vitro Chrysanthemum indicum L. in Da Lat - Lam Dong

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/14441 ◽

2021 ◽

Vol 19 (1) ◽

pp. 175-184

Author(s):

Phan Xuan Huyen ◽

Truong Ngoc Thao Vy ◽

Nguyen Thi Phuong Hoang ◽

Nguyen Thi Thanh Hang ◽

Dinh Van Khiem

Keyword(s):

Plant Height ◽

Shoot Regeneration ◽

Activated Charcoal ◽

In Vitro Propagation ◽

Ms Medium ◽

Coconut Fiber ◽

Regeneration Rate ◽

Shoot Height ◽

Chrysanthemum Indicum

Chrysanthemum indicum plant is a valuable and beneficial herb for human health. In this paper, we present the results of in vitro propagation of this plant in Da Lat. The results showed that MS medium supplemented with 25 g/l sucrose, 9 g/l agar, pH 5.8 was the best for shoot regeneration and growth (shoot height was 2.41 - 2.47 cm, 1 shoot/explant). MS media supplemented with BA (0.1, 0.5, 1, 1.5, 2 mg/l), Kinetin (0.1, 0.5, 1, 1.5, 2 mg/l ) and TDZ (0.1, 0.5, 1 mg/l) were unsuitable for shoot regeneration and growth of C. indicum. The regeneration and growth of shoots on the medium supplemented with 1 g/l of activated charcoal was better than (plant height of 3.45 cm) medium without activated charcoal (plant height of 2.46 cm). MS medium supplemented with 0, 0.1, 0.5, 1 mg/l IBA, 25 g/l sucrose, 9 g/l agar, pH 5,8 were suitable for in vitro root regeneration, rate of root formation was 100%. Coconut fiber powder was the best substrate to transfer the plantlets to the greenhouse, with survial rate of 100%, plantlets grew well. Nitrophoska irrigation was the best for the growth and development of C. indicum cultivated on coconut fiber powder under greenhouse conditions (plant height of 61.70 cm, 100.80 flowers/tree, flower diameter of 1.68 cm, fresh weight of 0.376 g/flower). The results also show that seedlings derived from tissue culture can be used to cultivate C. indicum and C. indicum grew well in the climate of Da Lat - Lam Dong and flowered all year round.

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The Effect of MS strength, pH and Sucrose Concentrations on In vitro Propagation of Okra (A belmoschus esculentus L.) from Shoot Tip Explants .

10.21203/rs.3.rs-153475/v1 ◽

2021 ◽

Author(s):

Girma Gebrehiwot Woldeyes ◽

Tileye Feyissa Senbeta ◽

Alelegne Yeshamebel Adugna ◽

Agegnehu Wasse Abegaz

Keyword(s):

Activated Charcoal ◽

Root Length ◽

Sucrose Concentration ◽

Shoot Multiplication ◽

Shoot Tips ◽

Leaf Number ◽

Root Number ◽

Inhibiting Effect ◽

Ph Level

Abstract BackgroundOkra (Abelmoschus esculentus) is belongs to the family Malvaceae and genus Hibiscus. Conventional propagation of okra in a large scale is limited due to bacterial, fungal, viral disease and seed dormancy. Hence, micropropagation offers great potential to increasing the commercial availability of okra. This study aimed to investigate the effect of MS strength, sucrose concentration, and pH on the in vitro propagation of A. esculentus from shoot tips. ResultsOkra seeds were sterilized with 70% alcohol, 30% (v/v) Berekina and 0.13% HgCl2 (w/v) for 5, 40 and 10 minutes respectively. The shoots were excised from in vitro germinated seedlings and transferred to full strength MS liquid medium containing 1mg/L BAP. The in vitro initiated shoot tips were transferred to different MS strength (1, ½, ⅓, ¼) medium, sucrose concentration (30 g/L, 20 g/L, 15 g/L, 10 g/L) and pH level (pH 6.6, 6.2, 5.8, 5.4, 5.0) that contained 1.5 mg/L BAP and 200 mg/L activated charcoal. For rooting, different MS strength (full, ½, ⅓, ¼), sucrose (30 g/L, 20 g/L, 15 g/L and 10 g/L) and pH (6.6, 6.2, 5.8, 5.4, 5.0) that contained 1mg/L IBA with 200 mg/L activated charcoal were used. Full strength MS medium containing 30 g/L sucrose at pH 5.8 produced the highest mean shoot number (7.93), mean leaf number (14.37) and mean length (6.4 cm) per explant. Similarly, at pH 6.2 and 6.6 maximum shoot mean number (6.53, 5.80), leaf number (12.77, 10.43) and shoot length (6.32, 5.82 cm) were produced respectively. Maximum mean root number (15.73) and root length (6.05 cm) were recorded at ½ MS strength. At pH 5.8, 6.2 and 6.6 maximum mean root number (1.77, 1.47, 1.80) and root length (1.38, 1.25, 1.39 cm) were obtained. After acclimatization, 86% of plants survived in greenhouse. ConclusionDecreasing the MS strength, sucrose concentration and pH level had an inhibiting effect on both shoot multiplication and root formation of okra. Besides increasing pH value showed inhibiting effect on shoot multiplication.

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Effect of activated charcoal and ascorbic acid on in vitro morphogenesis and o-dihydroxyphenols content in Paphiopedilum insigne

Horticultural Science ◽

10.17221/68/2020-hortsci ◽

2022 ◽

Author(s):

Monika Poniewozik ◽

Marzena Parzymies ◽

Paweł Szot

Keyword(s):

Ascorbic Acid ◽

Phenolic Compounds ◽

Activated Charcoal ◽

Root Formation ◽

Multiplication Rate ◽

Ms Medium ◽

In Vitro Morphogenesis ◽

Half Strength ◽

Positive Effect

Phenolic compounds limit micropropagation of many orchids in vitro. The aim of the study was to estimate the effect of activated charcoal (AC);1, 2 or 4 g/L) or ascorbic acid (AA; 10, 20 or 30 mg/L) added to the half strength MS medium on the growth and o-dihydroxyphenols content in Paphiopedilum insigne in vitro. A positive effect of AC on the shoot and root formation has been found. The highest multiplication rate (5.6 shoots/explant) and rooting frequency were obtained on medium containing 2 g/L of AC. However, AC reduced the leaf number as compared to the control. The lowest content of o-dihydroxyphenols was marked in Paphiopedilum insigne leaves when the shoots were grown on medium with 10 mg/L AA, followed by AC at 1 or 2 g/L.

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