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New basal media for half-anther culture of Anthurium andreanum Linden ex André cv. Tropical

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Abstract

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.

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References

  • Arzate-Fernández AM, Nakazaki T, Yamagata H, Tanisaka T (1997) Production of double haploid plants from Lilium longiflorum Thunb. anther culture. Plant Sci 123:179–187

    Article  Google Scholar 

  • Atak Ç, Çelik Ö (2009) Micropropagation of Anthurium andraeanum from leaf explants. Pak J Bot 41(3):1155–1161

    CAS  Google Scholar 

  • Bejoy M, Sumitha VR, Anish NP (2008) Foliar regeneration in Anthurium andreanum Hort. cv. Agnihothri Biotech 7(1):134–138

    CAS  Google Scholar 

  • Bohuš O, Dedičová B, Hricová A, Šamaj J, Pretrová A (2004) Flax anther culture: effect of genotype, cold treatment and media. Plant Cell Tissue Organ Cult 79(2):233–238

    Article  Google Scholar 

  • Bouman H, Tiekstra A (2001) Mineral nutrition in tissue culture: influence on propagation and quality of the plantlets. In: Horst WJ et al (eds) Plant nutrition—food Security and sustainability of agro-ecosystems. Kluwer, The Netherlands, pp 316–317

    Google Scholar 

  • Cao H, Biswas MK, Lu Y, Amar MH, Tong Z, Xu Q, Juan Xu J, Guo W, Deng X (2010) Doubled haploid callus lines of Valencia sweet orange recovered from anther culture. Plant Cell Tissue Organ Cult. Published online: 17 Oct 2010

  • Cardi T, Iannamico V, D’ambrosio F, Filippone E, Lurquin PF (1993) In vitro regeneration and cytological characterization of shoots from leaf explants of three accessions of Solanum commersonii. Plant Cell Tissue Organ Cult 34(1):107–114

    Article  Google Scholar 

  • Cellárová E, Rychlová M, Seidelová A, Honriv R (2004) Comparison of mitotic activity and growth in two long term callus cultures of Matricaria recutita L. Acta Biotechnol 10(3):245–251

    Article  Google Scholar 

  • Chée R, Pool RM (1987) Improved organic media constituents for in vitro shoot multiplication of Vitis. Sci Hort 32:85–95

    Article  Google Scholar 

  • Chen FC, Kuehnle AR, Sugii N (1997) Anthurium roots for micropropagation and Agrobacterium-mediated gene transfer. Plant Cell Tissue Organ Cult 49:71–74

    Article  CAS  Google Scholar 

  • Chiancone B, Tassoni A, Bagni N, Germana MA (2006) Effect of polyamines on in vitro anther culture of Citrus clementina Hort. ex Tan. Plant Cell Tissue Organ Cult 87:145–153

    Google Scholar 

  • Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY, Bi FI (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sinica 18:659–668

    Google Scholar 

  • Cristea V, Brummer AT, Jarda L, Miclăuş M (2010) In vitro culture initiation and phytohormonal influence on Dianthus henteri—a Romanian endemic species. Rom Biotechnol Lett 15(1):25–33

    CAS  Google Scholar 

  • Darnaedi D (1991) Informasi Kromosom. Pelatihan Sitogenetika. PAU Ilmu Hayat, IPB, 5 Nov 1991. Bogor

  • Doi H, Takahashi R, Hikage T, Takahata Y (2010) Embryogenesis and doubled haploid production from anther culture in gentian (Gentiana triflora). Plant Cell Tissue Organ Cult 102:27–33

    Article  Google Scholar 

  • Dolezel J, Novak FJ (1984) Cytogenetic effect of plant tissue culture medium with certain growth substances on Allium sativum L. meristem root tip cells. Biol Plant 26(4):293–298

    Article  CAS  Google Scholar 

  • Eeckhaut T, Werbrouck S, Dendauw J, van Bockstaele E, Debergh P (2001) Induction of homozygous Spatiphyllum wallisii genotypes through gynogenesis. Plant Cell Tissue Organ Cult 67:181–189

    Article  CAS  Google Scholar 

  • Fras A, Maluszynska J (2004) The correlation between the chromosome variation in callus and genotype of explants of Arabidopsis thaliana. Genetica 121(2):145–154

    Article  PubMed  Google Scholar 

  • Fu XP, Yang SH, Bao MZ (2008) Factors affecting somatic embryogenesis in anther cultures of Chinese pink (Dianthus chinensis L.). In Vitro Cell Dev Biol Plant 44:194–202

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Gantait S, Mandal N, Bhattacharyya S, Das PK (2008) In vitro mass multiplication with pure genetic identity in Anthurium andreanum Lind. Plant Tissue Cult Biotech 18(2):113–122

    Google Scholar 

  • George EF, Hall MA, De Klerk GJ (2007) Plant propagation by tissue culture. In: The background. exegetic, vol 1, 3rd edn, Basingstone

  • Gorecka K, Krzyzanowska D, Kiszczak W, Kowalska U (2009) Plant regeneration from carrot (Daucus carota L.) anther culture derived embryos. Acta Physiol Plant 31:1139–1145

    Article  Google Scholar 

  • Guo YD, Sewon P, Pulli S (1999) Improved embryogenesis from anther culture and plant regeneration in timothy. Plant Cell Tissue Organ Cult 57:85–93

    Article  Google Scholar 

  • Hamidah M, Karim AGA, Debergh P (1997) Somatic embryogenesis and plant regeneration in Anturium scherzerianum. Plant Cell Tissue Organ Cult 48:189–193

    Article  Google Scholar 

  • Han DS, Niimi Y, Nakano M (1997) Regeneration of haploid plants from anther cultures of the Asiatic Irbid lily ‘Connecticut King’. Plant Cell Tissue Organ Cult 47:153–158

    Article  Google Scholar 

  • Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circ Univ Calif Agric Exp Station Berkley, p 347

  • Huang B, Keller WA (1989) Microspore culture technology. J Tissue Cult Methods 12:171–178

    Article  Google Scholar 

  • Ishizaka H (1998) Production of microspore-derived plants by anther culture of an interspecific F1 hybrid between Cyclamen persicum and C. purpurascens. Plant Cell Tissue Organ Cult 54:21–28

    Article  Google Scholar 

  • Jahan MT, Islam MR, Khan R, Mamun ANK, Ahmed G, Hakim L (2009) In vitro clonal propagation of anthurium (Anthurium andraeanum L.) using callus culture. Plant Tissue Organ Cult Biotech 19(1):61–69

    Google Scholar 

  • Jin S, Mushke R, Zhu H, Tu L, Lin Z, Zhang Y, Zhang X (2008) Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers. Plant Cell Rep 27(8):1303–1316

    Article  PubMed  CAS  Google Scholar 

  • Joseph D, Martin KP, Madassery J, Philip VJ (2003) In vitro propagation of three commercial cut flower cultivars of Anthurium andraeanum L. Hort. Indian J Exp Biol 41:154–159

    Google Scholar 

  • Khorami R, Safarnejad A (2011) In vitro selection of Foeniculum vulgare for salt tolerance. Not Sci Biol 3(2):90–97

  • Knudson K (1946) A new nutrient solution for germination of orchid seeds. Am Orchid Soc Bull 15:214–217

    CAS  Google Scholar 

  • Koleva-Gudeva LR, Spasenoski M, Trajkova F (2007) Somatic embryogenesis in pepper anther culture: the effect of incubation treatments and different media. Sci Hort 111(2):114–119

    Article  CAS  Google Scholar 

  • Kozik EU, Nowak R, Kłosiñska U, Górecka K, Krzyżanowska D, Górecki R (2002) Morphological diversity of androgenic carrot plants. J Appl Genet 43(1):49–53

    PubMed  Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue culture. Physiol Plant 18:100–127

    Article  CAS  Google Scholar 

  • Lloyd G, McCown B (1981) Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Comb Proc Int Plant Propag Soc 30:421–427

    Google Scholar 

  • Maira O, Alexander N, Vargas TE (2009) Micropropagation and organogenesis of Anthurium andreanum Lind cv Rubrun. Plant Sci 585:3–14

    Google Scholar 

  • Martin KP, Joseph D, Madassery J, Philip VJ (2003) Direct shoot regeneration from lamina explants of two commercial cut flower cultivars of Anthurium andraeanum L. Hort. In Vitro Cell Dev Biol Plant 39(5):500–504

    Article  Google Scholar 

  • Metwally EI, Moustafa SA, El-Sawy BI, Shalaby TA (1998) Haploid plantlets derived by anther culture of Cucurbita pepo. Plant Cell Tissue Organ Cult 52:171–176

    Article  CAS  Google Scholar 

  • Miller LR, Murashige T (1976) Tissue culture propagation of tropical foliage plants. In Vitro 12(12):797–813

    Article  PubMed  CAS  Google Scholar 

  • Morard P, Henry M (1998) Optimization of the mineral composition of in vitro culture media. J Plant Nutr 21(8):1565–1576

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Murashige T, Tucker DPH (1969) Growth factor requirement of citrus tissue culture. In: International citrus symposium, 1, Riverside, Proceedings Riverside, vol 3. University of California, pp 1155–1169

  • Niedz RP, Evens TJ (2007) Regulating plant tissue growth by mineral nutrition. In Vitro Cell Dev Biol Plant 43:370–381

    Article  CAS  Google Scholar 

  • Nitsch JP, Nitsch C (1969) Haploid plant from pollen grains. Science 169:85

    Article  Google Scholar 

  • Nomizu T, Niimi Y, Han DS (2004) Haploid plant regeneration via embryogenesis from anther cultures of Hepatica nobilis. Plant Cell Tissue Organ Cult 79:307–313

    Article  Google Scholar 

  • Nowak BK, Miczyński K, Hudy L (2007) The effect of total inorganic nitrogen and the balance between its ionic forms on adventitious bud formation and callus growth of ‘Węgierka Zwykła’ plum (Prunus domestica L). Acta Physiol Plant 29(5):479–484

    Article  CAS  Google Scholar 

  • Ockendon DJ (2008) The ploidy of plants obtained from anther culture of cauliflowers (Brassica oleracea var. botrytis). Ann Appl Biol 113(2):319–325

    Article  Google Scholar 

  • Quoirin M, Lepoivre P (1977) Improved medium for in vitro culture of Prunus sp. Acta Hort 78:437–442

  • Rachmawati F (2005) Kultur anther pada anthurium (Anthurium andreanum Linden ex André). Thesis. Departemen Agronomi dan Hortikultura. Fakultas Pertanian. Institut Pertanian Bogor, 133 Halaman, Indonesia

  • Rimberia F, Adaniya S, Etoh T, Ishimine Y (2006) Sex and ploidy of anther culture derived papaya (Carica papaya L.) plants. Euphytica 149(1–2):53–59

    Article  Google Scholar 

  • Rodrigues IR, Forte BC, Olivera JMS, Mariath JEA, Bodanese-Zanettini MH (2004) Effects of light conditions and 2, 4-d concentration in soybean anther culture. Plant Growth Regul 44:125–131

    Article  CAS  Google Scholar 

  • Saji KV, Sujatha M (1998) Embryogenesis and plant regeneration in anther culture of sunflower (Helianthus anuus L.). Euphytica 103:1–7

    Article  Google Scholar 

  • Sayem MA, Maniruzzaman M, Siddique SS, Al-Amin M (2010) In vitro shoot regeneration through anther culture of Brassica spp. Bangladesh J Agric Res 35(2):331–341

    Google Scholar 

  • Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50:199

    Article  CAS  Google Scholar 

  • Seguí-Simarro JM, Nuez F (2007) Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers. J Exp Bot 58(5):1119–1132

    Google Scholar 

  • Shtereva LA, Zagorska NA, Dimitrov BD, Kruleva MM, Oanh HK (1998) Induced androgenesis in tomato (Lycopersicon esculentum Mill). II. Factors affecting induction of androgenesis. Plant Cell Rep 18:312–317

    Article  CAS  Google Scholar 

  • Supena EDJ, Suharsono S, Jacobsen E, Custers JBM (2006) Successful development of a shed-microspore culture protocol for doubled haploid production in Indonesian hot pepper (Capsicum annuum L.). Plant Cell Rep 25:1–10

    Article  PubMed  CAS  Google Scholar 

  • Sutan AN, Popescu A, Isac V (2010) In vitro culture medium and explant type effect on callogenesis and shoot regeneration in two genotypes of ornamental strawberry. Rom Biotech Lett 15(2):12–18

    Google Scholar 

  • Szarejko I, Falk DE, Janusz A, Nabialkowska D (1997) Cytological and genetic evaluation of anther culture-derived doubled haploids in barley. J Appl Genet 38(4):437–452

    Google Scholar 

  • Teixeira da Silva JA, Tanaka M (2006) Embryogenic callus, PLB and TCL paths to regeneration in hybrid Cymbidium (Orchidaceae). J Plant Growth Reg 25(3):203–210

    Article  CAS  Google Scholar 

  • Teixeira da Silva JA, Yam T, Fukai S, Nayak N, Tanaka M (2005) Establishment of optimum nutrient media for in vitro propagation of Cymbidium Sw. (Orchidaceae) using protocorm-like body segments. Propag Ornam Plants 5(3):129–136

  • Teng WL (1997) Regeneration of anthurium adventitious shoots using liquid or raft culture. Plant Cell Tissue Organ Cult 49:153–156

    Article  Google Scholar 

  • Thengane SR, Joshi MS, Khuspe SS, Mascarenhas AE (1994) Anther culture in Helianthus annuus L., influence of genotype and culture conditions on embryo induction and plant regeneration. Plant Cell Rep 13:222–226

    Article  CAS  Google Scholar 

  • Thimijan RW, Heins HD (1982) Photometric, radiometric, and quantum light units of measure: a review of procedures for interconversion. HortSci 18:818–822

    Google Scholar 

  • Vacin EF, Went EW (1949) Some pH changes in nutrient solution. Bot Gaz 110:605–613

    Article  CAS  Google Scholar 

  • Vargas TE, Mejias A, Oropeza M, de Garcia E (2004) Plant regeneration of Anthurium andraeanum L. cv. Rubrun. Electron J Biotechnol 7(3):5

    Google Scholar 

  • Viégas J, da Rocha MTR, Ferreira-Moura I, Corrêa MGS, da Silva JB, dos Santos NC, Teixeira da Silva JA (2007) Anthurium andraeanum (Linden ex André) culture: in vitro and ex vitro. Floric Ornam Biotechnol 1(1):61–65

  • Wang J, Bao MZ (2007) Plant regeneration of pansy (Viola wittrockiana) ‘Caidie’ via petiole-derived callus. Sci Hort 111(3):266–270

    Article  CAS  Google Scholar 

  • Westfall PH, Tobias RD, Rom D, Wolfinger RD, Hochberg Y (1999) Multiple comparisons and multiple tests: using the SAS system. SAS Publishing, SAS Institute Inc, Cary

    Google Scholar 

  • White PR (1963) The cultivation of animal and plant cells. Ronald Press, New York

    Google Scholar 

  • Winarto B (2006) The effect of explant types and culture media on the in vitro regeneration of adventitious shoots of rose. J Agrotrop 11(2):67–73

    Google Scholar 

  • Winarto B (2007) Response of axillary and adventitious shoot formation of in vitro culture of Anthurium. J Hortic 17(1):17–25

    Google Scholar 

  • Winarto B (2009) Androgenesis: a breakthrough effort for preparing haploid or double-haploid plants in Anthurium. PhD Dissertation. Department of Agronomy and Horticulture, Faculty of Agriculture. Bogor Agriculture Institute, 235 pp

  • Winarto B, Mattjik NA (2009a) Response of Anthurium andreanum Linden ex André cv. Carnaval anther in medium with various combinations of plant growth regulator concentrations. J Agron Indonesia 37(2):138–144

    Google Scholar 

  • Winarto B, Mattjik NA (2009b) Studies on anther culture of local accessions of Anthurium. J Hortic 19(4):386–395

    Google Scholar 

  • Winarto B, Rachmawati F (2007) Preliminary study of anther culture of Anthurium. J Hortic 17(2):127–137

    Google Scholar 

  • Winarto B, Mattjik NA, Purwito A, Marwoto B (2010a) Improvement of selected induction culture media on callus induction in anther culture of Anthurium and a histological study on its callus formation. J Nat Indonesia 12(2):93–101

    Google Scholar 

  • Winarto B, Mattjik NA, Teixeira da Silva JA, Purwito A, Marwoto B (2010b) Ploidy screening of anthurium (Anthurium andreanum Linden ex André) regenerants derived from anther culture. Sci Hortic 127:86–90

    Article  Google Scholar 

  • Winarto B, Rachmawati F, Pramanik D, Teixeira da Silva JA (2011) Morphological and cytological diversity of regenerants derived from Anthurium half-anther culture. Plant Cell Tissue Organ Cult 105(3):375–382

    Article  Google Scholar 

  • Yu YX, Liu L, Liu JX, Wang J (2009) Plant regeneration by callus-mediated protocorm-like body induction of Anthurium andraeanum Hort. Agric Sci China 8(5):572–577

    Article  CAS  Google Scholar 

  • Zhao FC, Nilanthi D, Yang YS, Wu H (2006) Anther culture and haploid plant regeneration in purple coneflower (Echinacea purpurea L.). Plant Cell Tiss Organ Cult 86:55–62

    Article  Google Scholar 

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Winarto, B., Rachmawati, F. & Teixeira da Silva, J.A. New basal media for half-anther culture of Anthurium andreanum Linden ex André cv. Tropical. Plant Growth Regul 65, 513–529 (2011). https://doi.org/10.1007/s10725-011-9622-x

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