In Vitro Regeneration through Direct Organogenesis from Protocorms of Oncidium tigrinum Llave & Lex. (Orchidaceae), an Endemic and Threatened Mexican Species

A protocol for in vitro propagation from protocorms of Oncidium tigrinum Llave & Lex., a threatened species distributed in Mexico and highly appreciated as an ornamental, was developed. Two different explants, entire protocorms and longitudinal halves of protocorms, were used. In addition, the effect of two different culture media, Murashige and Skoog (MS) and modified Knudson (KCm), supplemented with N6-benzyladenine (BA) (0, 0.5, 1, 2, 3, and 5 mg·L−1) and/or α-naphthaleneacetic acid (NAA) at 0, 0.1, and 0.5 mg·L−1 was investigated. Adventitious shoot formation by direct organogenesis was obtained in all treatments; in some cases, the formation of protocorm-like bodies was induced. Shoot formation was greater for entire protocorms; the best treatment was MS medium containing at BA 1 to 2 mg·L−1 in combination with at NAA 0.1 mg·L−1. The average height of shoots was three times greater in MS medium than in KCm medium. Subculturing individual shoots in MS medium without plant growth regulators, but with 1 g·L−1 activated charcoal, allowed further development and rooting. An ex vitro survival rate of almost 100% was achieved. This study represents a comprehensive application for propagation, conservation, and sustainable use of this valuable natural resource.

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In vitro studies on Dendrobium fimbriatum Hk.F – An endangered orchid

Journal of Applied and Advanced Research ◽

10.21839/jaar.2019.v4i4.69 ◽

2019 ◽

pp. 100-103

Author(s):

T. Ramesh ◽

P. Renganathan ◽

M. Prabhakaran

Keyword(s):

Culture Media ◽

Adventitious Shoot ◽

Shoot Formation ◽

Direct Organogenesis ◽

Naphthalene Acetic Acid ◽

Average Height ◽

Ms Medium ◽

Adventitious Shoot Formation ◽

Dendrobium Fimbriatum

A protocol for in vitro propagation from protocorms of Dendrobium fimbriatum a distributed in all over world and highly appreciated as an ornamental, was developed. Two different explants, entire protocorms and longitudinal halves of protocorms, were used. In addition, the effect of two different culture media, Murashige and Skoog (MS) and modified Knudson (KCm), supplemented with N6- benzyladenine (BA) (0, 0.5, 1, 2, 3, and 5 mg_L–1) and/or a-naphthalene acetic acid (NAA) at 0, 0.1, and 0.5 mg_L–1 was investigated. Adventitious shoot formation by direct organogenesis was obtained in all treatments; in some cases, the formation of protocorm like bodies was induced. Shoot formation was greater for entire protocorms; the best treatment was MS medium containing at BA 1 to 2 mg_L–1 in combination with at NAA 0.1 mg_L–1. The average height of shoots was three times greater in MS medium than in KC m medium. Sub culturing individual shoots in MS medium without plant growth regulators, but with 1 g_L–1 activated charcoal, allowed further development and rooting. An ex vitro survival rate of almost 100% was achieved. This study represents a comprehensive application for propagation, conservation, and sustainable use of this valuable natural resource.

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In Vitro Regeneration from Longitudinal Sections of Seedlings of Beaucarnea purpusii Rose, an Endemic and Endangered Species

HortScience ◽

10.21273/hortsci.51.3.279 ◽

2016 ◽

Vol 51 (3) ◽

pp. 279-284 ◽

Cited By ~ 2

Author(s):

María del Carmen Vadillo-Pro ◽

Luis Hernández-Sandoval ◽

Guadalupe Malda-Barrera ◽

María Luisa Osorio-Rosales ◽

Martín Mata-Rosas

Keyword(s):

Liquid Culture ◽

Culture Media ◽

In Vitro Regeneration ◽

Sustainable Use ◽

Adventitious Shoot ◽

Shoot Formation ◽

Direct Organogenesis ◽

Adventitious Shoot Formation ◽

Efficient Alternative

The present study establishes an efficient protocol for in vitro propagation from longitudinal sections of seedlings of Beaucarnea purpusii, a threatened and highly appreciated ornamental species. The effect of three cytokinins: N6-benzyladenine (BA), kinetin (Kin), and thidiazuron, 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (TDZ), in semisolid media and three different concentrations, as well as the effect of BA and TDZ pulses at higher concentrations in liquid culture medium, were investigated. Adventitious shoot formation by direct organogenesis was observed from all treatments. Additionally, adventitious shoot formation was recorded from the leaves of the new shoots; this particular response was exclusive to treatments supplemented with TDZ. In the experiment using semisolid culture media, the highest means of shoots per explant were obtained from treatments containing TDZ, particularly at a concentration of 0.45 μm (25.8 shoots per explant). For the pulses experiment, the liquid culture media supplemented with TDZ at 22.35 µm for 24 hours and 136.21 µm for 96 hours, induced a mean of 3.9 and 3.3 shoots per explant, respectively. Subculturing individual shoots on MS and half-strength MS (1/2MS) media, both supplemented with activated charcoal at 1 g·L−1, induced rooting in 85% to 95% of shoots. A survival rate of 100% under greenhouse conditions was achieved. The results of this study provide an efficient alternative for mass propagation of B. purpusii and may also contribute to the conservation and sustainable use of this valuable natural resource.

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In vitro Regeneration of the Medicinal Plant, Plumbago zeylanica L. with Reference to a Unique Population in Maruthamalai, the Western Ghats, India

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v18i2.3648 ◽

1970 ◽

Vol 18 (2) ◽

pp. 173-179 ◽

Cited By ~ 1

Author(s):

T. Mallikadevi ◽

P. Senthilkumar ◽

S. Paulsamy

Keyword(s):

Medicinal Plant ◽

In Vitro Regeneration ◽

Basal Medium ◽

Adventitious Shoot ◽

Shoot Formation ◽

Garden Soil ◽

Plumbago Zeylanica ◽

Adventitious Shoot Formation ◽

The Western Ghats

The in vitro regeneration of Plubago zeylanica exhibited that the callus was initiated in the basal medium containing BAP, NAA, 2, 4-D, and IBA. The high amount (90%) of organic calli was induced in the basal medium supplemented with 2, 4-D, alone at 2.0 mg/l. In the subculture the adventitious shoot formation was prominently higher (83%) in the basal medium containing BAP, and NAA at 3.5 and 0.3 mg/l, respectively. IAA (1.0 mg/l)effectively produced higher percen-tage (90) of roots and root growth. After sequential hardening, survivability rate was observed to be significantly higher (80%) in the hardening medium containing garden soil, sand and vermicompost in the ratio of 1 : 1 : 1 by volume under greenhouse condition. Key words: Plumbago zeylanica, In vitro regeneration, Medicinal plant D.O.I. 10.3329/ptcb.v18i2.3648 Plant Tissue Cult. & Biotech. 18(2): 173-179, 2008 (December)

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882 PB 276 IN VITRO PROPAGATION AND CHIMERAL TRAITS OF Cryptanthus `Marian Oppenheimer' (WIDE LEAF CLONE)

HortScience ◽

10.21273/hortsci.29.5.560c ◽

1994 ◽

Vol 29 (5) ◽

pp. 560c-560

Author(s):

Yong Cheong Koh ◽

Fred T. Davies

Keyword(s):

In Vitro Propagation ◽

Callus Formation ◽

Adventitious Shoot ◽

Shoot Formation ◽

Induction Medium ◽

Ms Medium ◽

Periclinal Chimera ◽

Adventitious Shoot Formation ◽

L1 And L2

The leaves of vegetative stolons of greenhouse grown Cryptanthus `Marian Oppenheimer' (wide leaf clone) were cultured in modified MS media to induce adventitious shoot formation via callus formation. The best callus induction medium was basal MS medium with 10 μM NAA, IBA and BA. Pure green (843), maroon (3), striped (2) and albino plantlets were obtained. Most of the albino plantlets were stunted, tightly clumped together and impossible to score. The medium which produced the highest average number of non-albino plantlets was basal MS medium with 0.3 μM NAA, IBA and BA All non-albino plantlets were rooted in MS medium with 5.4 μM NAA and transplanted ex vitro with a survival rate of 96.7%. The maroon plantlets became green two weeks after transplanting. Histological studies revealed that C. `Marian Oppenheimer' (wide leaf clone) has two tunicas (L1 and L2) and a corpus (L3). Callus on the leaf explant arose mainly from the L2 and L3. Apparently C. `Marian Oppenheimer' (wide leaf clone) is a GWG periclinal chimera.

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883 PB 279 REGENERATING ADVENTITIOUS PLANTS FROM IN VITRO CULTURE OF LIATRIS SPICATA (L.) WILLD. COTYLEDONS

HortScience ◽

10.21273/hortsci.29.5.560d ◽

1994 ◽

Vol 29 (5) ◽

pp. 560d-560

Author(s):

Dennis P. Stimart ◽

John C. Mather

Keyword(s):

In Vitro Culture ◽

Adventitious Shoots ◽

Adventitious Shoot ◽

Shoot Formation ◽

Ms Medium ◽

Ex Vitro ◽

Adventitious Shoot Formation ◽

Murashige Skoog ◽

Micropropagated Plants

Cotyledons from developing embryos 6 to 8 weeks old of Liatris spicata (blazing star) were cultured on Murashige-Skoog (MS) medium containing 0, 0.4, 4.4, and 44.4 μ M benzyladenine (BA) or 0, 0.2, 2.2, and 22.2 μ M thidiazuron (TDZ) to induce adventitious shoot formation. The highest percent of cotyledons forming shoots with highest shoot counts was on medium containing 2.2 μ M TDZ. Vitreous shoots formed on medium with 22.2 μ M TDZ. Callus derived from cotyledons and cultured on medium containing 4.44 μ M BA or 2.2 μ M TDZ formed adventitious shoots with highest shoot counts on 4.44 μ M BA. Adventitious shoots derived from cotyledons and callus were rooted on MS medium with 5.0 μ Mindole-3-butyric acid, acclimatized and grown ex vitro. All micropropagated plants appeared similar to each other.

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Regenerating Adventitious Shoots from in Vitro Culture of Liatris spicata (L.) Willd. Cotyledons

HortScience ◽

10.21273/hortsci.31.1.154 ◽

1996 ◽

Vol 31 (1) ◽

pp. 154-155

Author(s):

Dennis P. Stimart ◽

John C. Mather

Keyword(s):

In Vitro Culture ◽

Adventitious Shoots ◽

Adventitious Shoot ◽

Shoot Formation ◽

Ms Medium ◽

Adventitious Shoot Formation ◽

Regenerated Plants ◽

Murashige And Skoog Medium ◽

Homogeneous Chemical

Cotyledons from developing 6- to 8-week-old embryos of Liatris spicata (L.) Willd. (blazing star) were cultured on Murashige and Skoog medium containing 0, 0.4, 4.4, or 44.4 μm BA or 0, 0.2, 2.2, or 22.2 μm TDZ to induce adventitious shoot formation. The highest percentage of cotyledons forming the most shoots was on medium containing 2.2 μm TDZ. Cotyledon-derived callus cultured on medium containing 4.4 μm BA formed ≈16 times more adventitious shoots than on 2.2 μm TDZ. Adventitious shoots derived from cotyledons or callus produced roots when placed on MS medium containing 5.0 μm IBA. Regenerated plants that flowered in the field appeared homogeneous. Chemical names used: N6-benzyladenine (BA), thidiazuron (TDZ), indole-3-butyric acid (IBA).

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Adventitious Shoot Formation on Excised Hypocotyls of Antirrhinum majus L. (Snapdragon) In Vitro

HortScience ◽

10.21273/hortsci.34.4.736 ◽

1999 ◽

Vol 34 (4) ◽

pp. 736-739

Author(s):

Kenneth R. Schroeder ◽

Dennis P. Stimart

Keyword(s):

Adventitious Shoot ◽

Shoot Formation ◽

Antirrhinum Majus ◽

Ms Medium ◽

Optimal Conditions ◽

Dominant Trait ◽

Continuous Darkness ◽

Adventitious Shoot Formation ◽

Murashige Skoog

Hypocotyls from Antirrhinum majus L. were excised at 2 weeks of age from seedlings grown under a 16-hour photoperiod or continuous darkness. Explants were cultured on modified Murashige-Skoog (MS) medium containing 0, 0.44, 2.22, 4.44, 8.88, or 44.4 μm BA to investigate adventitious shoot formation. Excised hypocotyls from eight commercial cultivars, three inbred lines, and an F1 hybrid between two of the inbreds were cultured on MS medium containing 2.22 μm BA to assess genotypic effects on adventitious shoot formation. The influence of seedling age was assessed by excising hypocotyls from seedlings at 6, 10, 14, 18, 22, 26, or 30 days. Optimal conditions for adventitious shoot formation on excised hypocotyls included: seedling growth in a lighted environment, use of hypocotyls from 10-day-old seedlings, and culture on medium containing 2.22 μm BA for 3 weeks. Under these conditions, up to a 5-fold improvement in number of shoots per hypocotyl over previous studies was achieved. Adventitious shoot formation was genotype-dependent and appeared to be a dominant trait. Chemical name used: N6-benzyladenine (BA).

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In Vitro Regeneration of Valencia-type Peanut (Arachis hypogaea L.) from Cultured Petiolules, Epicotyl Sections and Other Seedling Explants1

Peanut Science ◽

10.3146/i0095-3679-19-2-5 ◽

1992 ◽

Vol 19 (2) ◽

pp. 82-87 ◽

Cited By ~ 25

Author(s):

Ming Cheng ◽

David C. H. Hsi ◽

Gregory C. Phillips

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Shoot Formation ◽

Direct Organogenesis ◽

Naphthaleneacetic Acid ◽

Regeneration System ◽

Arachis Hypogaea L ◽

Wide Range ◽

Cultivated Peanut

Abstract This study evaluated plant development via direct organogenesis from in vitro-cultured young seedling tissues of cultivated peanut, especially the valencia-type peanut. Complete plants were regenerated from in vitro-cultured petiolule-with-blade-attached explants, leaflet segments, and epicotyl andpetiole sections. Multiple shoots arose on Murashige and Skoog medium (MS) supplemented with 6-benzylaminopurine (BA) (5–25 mg/L) plus 1-naphthaleneacetic acid (NAA) (0.5–3 mg/L). After 30 d culture on 25 mg/L BA + 1 mg/L NAA, 1.6 buds or shoots/explant were regenerated from the petiolule-with-blade-attached explants. Comparable numbers of shoots were obtained from epicotyl sections of the first node region of the seedling after 60 d culture using 10 mg/L BA + 1 mg/L NAA. Leaflet segments and petiole sections were less responsive for shoot formation. Excised shoots developed roots in vitro upon transfer for 15 d to MS medium supplemented with NAA at 1 mg/L. Plantlets were transferred to soil and grown in a greenhouse to maturity. A wide range of cultivated peanut genotypes was evaluated for organogenic responsiveness, using the petiolule-with-blade-attached explant source. Only valencia-type cultivars, or a hybrid derivative with a Valencia background, were responsive with this regeneration system.

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Improved Adventitious Shoot Production in White Ash

HortScience ◽

10.21273/hortsci.30.4.874c ◽

1995 ◽

Vol 30 (4) ◽

pp. 874C-874

Author(s):

Sharon A. Bates ◽

John E. Preece ◽

John H. Yopp

Keyword(s):

Adventitious Shoot ◽

Shoot Elongation ◽

Shoot Formation ◽

Separate Experiment ◽

Ms Medium ◽

Adventitious Shoot Formation ◽

White Ash ◽

Shoot Initiation ◽

Shoot Production

To increase adventitious shoot formation, we investigated the effects of the number of weeks on medium with high levels of plant growth regulators and seedcoat removal. Dissected white ash seeds were placed on a solidified MS medium containing 10 μM TDZ and 1 μM 2,4-D (shoot initiation medium). After 2, 3, or 4 weeks in vitro, explants were transferred to shoot elongation medium (3 μM TDZ, 1 μM BA, and 1 μM IBA). After 12 weeks, the greatest number (1.8) and longest shoots (18.7 mm) were in cultures incubated on the shoot formation medium for 3 weeks. In a separate experiment, dissected seeds were placed on shoot formation medium. Seedcoats were removed after 10 days in vitro. Explants were transferred to shoot elongation medium after 4 weeks in vitro. There were more shoots (2.5) on 12-week-old explants without seedcoats than on explants with seedcoats (0.9). This result may be related to inhibitors in the testa.

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Establishment of Cell Suspension Culture and Plant Regeneration in Abrus precatorius L., a Rare Medicinal Plant

Notulae Scientia Biologicae ◽

10.15835/nsb417187 ◽

2012 ◽

Vol 4 (1) ◽

pp. 86-93 ◽

Cited By ~ 6

Author(s):

Mohammad Serajur RAHMAN ◽

Mohammad Abdul Bari MIAH ◽

Mohammad Shahadat HOSSAIN ◽

Ahmad Humayan KABIR ◽

Mohammad Motiur RAHMAN

Keyword(s):

In Vitro Regeneration ◽

Shoot Formation ◽

Naphthaleneacetic Acid ◽

Root Induction ◽

Ms Medium ◽

Liquid Media ◽

Indolebutyric Acid ◽

Abrus Precatorius ◽

Isolated Cell

A new protocol has been developed for cell culture and in vitro regeneration of Abrus precatorius that holds enormous potentiality for preparation of medicines. In vitro grown calli were cultured in Murashige and Skoog (MS) liquid media in agitated condition fortified with 0.5 mg/l 6-Benzylaminopurine. Growth curve of cells revealed that the cells continued to grow until 12 days of culture and got the highest peak from day 6-8. Isolated cell was found to produce highest 8.2% calli when suspended on MS medium supplemented with 0.5 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. Callus derived from single cell produced highest number of embryo (25-28%) cultured on MS medium fortified with 2.0 mg/l 6-Benzylaminopurine and 0.2 mg/l 1-Naphthaleneacetic acid. The bipolar embryos were selected and optimum shoot formation was recorded on MS medium supplemented with 2.0 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. The optimum root induction was noticed in MS medium supplemented with 1.0 mg/l 3-Indolebutyric acid. Rooted plantlets were successfully transferred to potting soil and acclimatized to outdoor conditions.

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