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Hossain M.M.,Chittagong University | Kant R.,Panjab University | Van P.T.,Ehime University | Van P.T.,Vietnam Institute of Agricultural Sciences | And 3 more authors.
Critical Reviews in Plant Sciences | Year: 2013

This review provides an informative and broad overview of orchid biotechnology, addressing several important aspects such as molecular systematics, modern breeding, in vitro morphogenesis, protoplast culture, flowering control, flower color, somaclonal variation, orchid mycorrhiza, pathogen resistance, virus diagnosis and production of virus-free plants, functional genomics, genetic transformation, conservation biotechnology and pharmaceutical biotechnology. This resource will provide valuable insight to researchers who are involved in orchid biology and floriculture, using biotechnology to advance research objectives. Producing an improved orchid through biotechnology for industrial purposes or to serve as a model plant for pure and applied sciences is well within reach and many of the current techniques and systems are already employed at the commercial production level. © 2013 Copyright Taylor and Francis Group, LLC.


Winarto B.,Indonesian Ornamental Crops Research Institute | Rachmawati F.,Indonesian Ornamental Crops Research Institute | Pramanik D.,Indonesian Ornamental Crops Research Institute | Teixeira da Silva J.A.,Kagawa University
Plant Cell, Tissue and Organ Culture | Year: 2011

Anthurium anther culture was successfully established using half-anthers as explants. Explants were cultured on Winarto-Teixeira basal medium (WT-1) containing 0.01 mg/l α-naphthalene acetic acid (NAA), 0.5 mg/l thidiazuron (TDZ), and 1.0 mg/l 6-benzylaminopurine (BAP), or on New Winarto-Teixeira basal medium (NWT-3) supplemented with 0.02 mg/l NAA, 1.5 mg/l TDZ, and 0.75 mg/l BAP for callus initiation. Regenerated calli produced multiple shoots on WT-1, which were then rooted in NWT-3 supplemented with 1% activated charcoal. Plantlets were acclimatized ex vitro using a mixture of burned rice husk, rice husk, and bamboo peat (1:1:1, v/v/v) as the potting medium. There was considerable morphological and cytological diversity of regenerants derived from anther culture, which are described in detail in this study. The callus cluster color ranged from green to light green and had a high regeneration capacity (7.3 and 4.8 shoots/callus cluster), light reddish-yellow callus showed moderate regeneration (2.6 shoots/callus cluster), while reddish-yellow callus had the lowest regeneration capacity (1.5 shoots/callus cluster). Morphological variations clearly observed in regenerants derived from this technique included alterations in plant size, peduncle length, spathe position compared to leaves, the type and number of buds, spathe and spadix color, and spadix length. There were also cytological variations in both in vitro and ex vitro regenerants of anther culture with 23-29% haploids, 5-10% aneuploids, 56-69% diploids, and 3-4% triploids. The results strengthen other studies in which the development of anther cultures, especially via callus formation, resulted in morphological and cytological alterations. These variations have been discussed to great length in this paper. © 2010 Springer Science+Business Media B.V.


Winarto B.,Indonesian Ornamental Crops Research Institute | Teixeira da Silva J.A.,Kagawa University
Scientia Horticulturae | Year: 2012

A new micropropagation protocol for leatherleaf fern (. Rumohra adiantiformis (G. Forst.) Ching) was successfully established using rhizomes as the donor explant, following appropriate sterilization. Murashige and Skoog (MS) medium containing 0.25. mg/l 2,4-dichlorophenoxy acetic acid (2,4-D), 0.2. mg/l α-naphthalene acetic acid (NAA), 1.0. mg/l 6-benzyl adenine (BA), and 0.5. mg/l thidiazuron (TDZ) with 30. g/l sucrose (IM-4) was the most appropriate medium for culture initiation. When entire rhizomes harvested from mother stock plants were cultured on a simple paper bridge containing liquid IM-4 medium, culture initiation improved with 3.4 rhizomes regenerated per sub-culture. The average multiplication rate of newly regenerated rhizomes increased to 5.7/rhizome on half-strength MS medium supplemented with 0.05. mg/l IAA, 0.25. mg/l BA, 0.5. mg/l Kin, 1. g/l activated charcoal and 20. g/l sucrose (MM-5). The level of multiplication peaked in the fifth subculture and retained high quality until the sixth subculture. From the seventh subculture onwards, the quality of regenerated fronds was reduced. The regenerated rhizomes rooted easily on MM-5 and could be acclimatized ex vitro with 97-100% survival. © 2012 Elsevier B.V.


Winarto B.,Indonesian Ornamental Crops Research Institute | Rachmawati F.,Indonesian Ornamental Crops Research Institute | da Silva J.A.T.,Kagawa University
Plant Growth Regulation | Year: 2011

A successful protocol for high frequency callus induction and plant regeneration from Anthurium andreanum Linden ex André cv. Tropical half-anthers is described. Different variables using Winarto and Teixeira and Murashige and Skoog basal media supplemented with several plant growth regulators [2,4-dichlorophenoxy acetic acid (0.1-1.0 mg/l), α-naphthalene acetic acid (0.01-0.2 mg/l), thidiazuron (0.5-2.0 mg/l), 6-benzylaminopurine (0.5-1.0 mg/l), and kinetin (0.5-1.0 mg/l)] were tested for their ability to induce high frequency callusing in half-anthers, indirect regeneration and rooting of shoots. Basal medium, as well as the combination and concentration of hormones applied, had a significant effect on callus formation, shoot regeneration and adventitious root formation. Winarto and Teixeira-1, an original basal medium containing 0.01 mg/l α-naphthalene acetic acid, 0.5 mg/l thidiazuron and 1.0 mg/l 6-benzylaminopurine was suitable for callus formation while an improved basal medium i. e., New Winarto-Teixeira-3 supplemented with 0.25 mg/l 2,4-dichlorophenoxy acetic acid, 0.02 mg/l α-naphthalene acetic acid, 1.5 mg/l thidiazuron and 0.75 mg/l 6-benzylaminopurine enhanced callus formation. High shoot regeneration and multiplication was also possible on New Winarto-Teixeira-3. Shoots formed a strong adventitious root system on New Winarto-Teixeira-3 containing 0.2 mg/l α-naphthalene acetic acid and 1.0 mg/l kinetin. Plantlets that varied in size and performance were successfully acclimatized and adapted to ex vitro conditions. Cytological analysis of 180 acclimatized-plantlets ex vitro revealed that 34 were haploid (n = 14-18), 15 aneuploid (n = 20-26), 126 diploid (n = 28-34) and 5 triploid (n = 45-57). The potential use of this protocol for developing half-anther culture of other Anthurium species or cultivars is discussed. © 2011 Springer Science+Business Media B.V.


Teixeira da Silva J.A.,P.O. Box 7 | Dobranszki J.,Debrecen University | Winarto B.,Indonesian Ornamental Crops Research Institute | Zeng S.,CAS South China Botanical Garden
Scientia Horticulturae | Year: 2015

Anthurium (. Anthurium spp.) is an ornamental that is widely appreciated around the world, primarily for its showy and colorful spadix. A successful tissue culture protocol for anthurium would allow for the mass clonal propagation of this plant to serve the floriculture pot-plant and cut-flower markets. Success has been achieved in anthurium tissue culture using several explant types, and the morphological and cytogenetic stability of such regenerants has also been tested. This review provides a detailed analysis of the conditions required for the successful culture of anthuriums in vitro. Besides micropropagation, this review also highlights selected current applications of in vitro anthurium biotechnology such as anther culture, polyploidy production, genetic transformation, and their importance in breeding work, synthetic seed technology and cryopreservation. © 2014 Elsevier B.V.


Teixeira da Silva J.A.,Miki cho Post Office | Dobranszki J.,Debrecen University | Zeng S.,CAS South China Botanical Garden | Winarto B.,Indonesian Ornamental Crops Research Institute | And 3 more authors.
Plant Cell, Tissue and Organ Culture | Year: 2015

Although Anthurium is an attractive and commercially popular ornamental plant, its genetic enhancement has lagged behind that of other ornamental crops. There are several agronomically important traits in need of improvement. These include novel flower colors and morphologies, increased shelf and vase lives, and resistance to bacterial blight (Xanthomonas axonopodis pv. dieffenbachiae), burrowing nematodes and abiotic stresses. The production of transgenic Anthuriums is critical because the conventional breeding of a cultivar with beneficial traits typically requires 8–10 years. This review evaluates the problems, challenges and progress associated with developing molecular markers for Anthurium and in genetically transforming this ornamental. Recent improvements have hastened the tissue culture and regeneration of transgenic plants primarily using Agrobacterium-based methods. Promoter analyses have focused on constitutive and tissue-enhanced gene expression with the green fluorescent protein being a more reliable reporter than β-glucuronidase. The development of molecular markers assists with phylogenetic analyses and PCR-based markers such as RAPD, SSR, SPAR, ISSR and AFLP can be used to differentiate cultivars and for genetic fingerprinting. The marker-assisted breeding of Anthurium will become more feasible once available data are used for association to specific traits. Work on the identification of quantitative trait loci for disease resistance and other traits such as flower colour is required and should incorporate new approaches, such as next-generation sequencing technologies. By highlighting the aforementioned bottlenecks and successes in this review, it is expected that the pace of Anthurium genetic improvement will increase with the multifaceted incorporation of focused priorities and new technology advancements. © 2015 Springer Science+Business Media Dordrecht


Teixeira da Silva J.A.,Miki cho post office | Winarto B.,Indonesian Ornamental Crops Research Institute | Dobranszki J.,Debrecen University | Zeng S.,CAS South China Botanical Garden
Acta Physiologiae Plantarum | Year: 2015

The use of anther culture to produce haploid plants is a useful breeding technique for Anthurium, but has only seen limited applications thus far. This review describes the advances achieved thus far, and provides practical tips and protocols that would allow researchers new to the field, or already experienced Anthurium tissue culture scientists, to establish a new field of research, or a novel way to derive haploid plants for breeding programs. Anther culture can result in considerable somaclonal variation, but this has translated into a novel way of inducing new color variants of the spathe, the most marketable part of this ornamental plant. Considering this variability in vitro, anther culture is most likely not the most suitable method for clonal propagation of important germplasm, but is certainly a viable method to induce new genetically stable leaf and flower color variants. © 2015, Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.


Pramanik D.,Indonesian Ornamental Crops Research Institute | Rachmawati F.,Indonesian Ornamental Crops Research Institute | Winarto B.,Indonesian Ornamental Crops Research Institute
Acta Horticulturae | Year: 2016

In vitro propagation study of Phalaenopsis 'Puspa Tiara Kencana' via somatic embryogenesis (SE) was successfully performed. Young leaf explants as the suitable explant produced embryogenic callus (EC) up to 19.9%, and 8.8% of them produced somatic embryos (SEs) directly on half-strength MS medium, supplemented with 0.5 mg L-1 of N6-benzylaminopurine (BAP) and 1 mg L-1 of thidiazuron (TDZ) within 38 days after initiation. EC readily developed on the leaf explants 3.25 g EC weight and 0.63 cm3 EC volume, on average, while half-strength MS medium supplemented with 0.5 mg L-1 BAP and 1 mg L-1 TDZ stimulated 0.7 g EC weight and 3.1 cm3 EC volume. EC grew and proliferated optimally in leaf explant with a callus growth of 2.2 g month-1, and on the New Phalaenopsis (NP) medium, supplemented with 1 mg L-1 TDZ and 0.5 mg L-1 BAP, with EC growth of 1.6 g month-1. After 12 weeks of culture, 20 germinated SEs per clump were successfully established on NP medium with 0.05 mg L-1 of BAP, while more vigorous and greener plantlets were determined on half-strength MS medium, containing 0.05 mg L-1 BAP.


Winarto B.,Indonesian Ornamental Crops Research Institute | da Silva J.A.T.,P. O. Box 7
In Vitro Cellular and Developmental Biology - Plant | Year: 2015

To improve the micropropagation of Dendrobium ‘Gradita 31’, coconut water (CW) and fertilizer media were used to enhance growth, proliferation, and germination of protocorm-like bodies (PLBs). PLBs formed when shoots approximately 0.4 cm long were cultured on semi-solid half-strength Murashige and Skoog (MS) basal medium containing 1 mg L−1 thidiazuron (TDZ) and 0.5 mg L−1 N6-benzyladenine (BA). After four rounds of 15-d subcultures, PLBs were initially proliferated in liquid half-strength MS medium supplemented with 0.3 mg L−1 TDZ and 0.1 mg L−1 α-naphthalene acetic acid (NAA) and subcultured monthly into fresh medium. When 15% (v/v) CW was added to half-strength MS medium, optimal growth and proliferation of PLBs resulted with the following principal findings: a maximum of 2.86 g total fresh weight (FW) of PLBs formed from 1 g of PLBs, an average of 0.46 g FW of PLBs added per subculture period, a total of 157.1 PLBs formed, an average of 25.3 PLBs added per subculture period, and a low percentage of PLB browning (20.7%). Half-strength MS medium containing 15% (v/v) CW with Rosasol® medium (liquid fertilizer; 1.5 g L−1 18 N:18P:18 K + 1.5 g L−1 25 N:10P:10 K + TE) resulted in a similar organogenic outcome with 25.5% PLB browning. PLBs germinated easily and rooted on Rosasol-supplemented plant growth regulator-free medium. In total, 180 Dendrobium ‘Gradita 31’ plantlets from all treatments were successfully acclimatized with 100% survival on a Cycas rumphii bulk substrate and grew well after repotting in a mixture of wood charcoal and C. rumphii bulk (1:1, v/v). © 2015, The Society for In Vitro Biology.


Winarto B.,Indonesian Ornamental Crops Research Institute | Rachmawati F.,Indonesian Ornamental Crops Research Institute | Setyawati A.S.,Indonesian Ornamental Research Station | da Silva J.A.T.,P. O. Box 7
Emirates Journal of Food and Agriculture | Year: 2015

Lisianthus (Eustoma grandiflorum (Raf.) Shinn) is an important ornamental commodity in South-East Asia. However, mass propagation of the plant at a commercial scale to satisfy market demands is faced by limited availability of high quality and uniform seedlings as planting material. Using different regeneration media and leaf explants for callus induction, regeneration, proliferation, root formation and acclimatization were studied. High callus induction and adventitious shoot formation were possible from leaf explants of E. grandiflorum 'White Lavender' cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/l thidiazuron (TDZ) and 0.3 mg/l α-naphthalene acetic acid (NAA), but high quality shoots (8.0) was established on MS medium containing 0.5 mg/l N6-benzyladenine (BA) and 0.002 mg/l NAA. In the same medium, adventitious shoots could be multiplied up to the fourth subculture at a rate of 1.74 which decreased to a 1.57 multiplication rate in subsequent subcultures. Shoots rooted easily on MS medium containing 0.1 mg/l BA and 0.02 mg/l NAA with 3.9 roots per shoot. The plantlets, which were successfully acclimatized in a mixture of burned-rice husk and organic manure (1:1, v/v) with 90% survival, grew well after repotting.

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