Morphological and cytological diversity of regenerants derived from half-anther cultures of anthurium

Abstract

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 a-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.

In vitro flowering and production of viable pollen of cucumber

“…Whole plants seem to be less susceptible to in vitro culture, and no differences in flower morphology were observed as compared to greenhouse-grown controls. Increasing incidence of morphological changes following regeneration after various cycles of in vitro culture of different explants has been reported in cucumber (Burza et al 1996;Pląder et al 1998), as well as in other plants (Breiman et al 1989;Hee et al 2007;Nikolić et al 2006;Winarto et al 2011). Ameha et al (1998) reported development of tendrils in regenerants from in vitro cultured whole seeds and seed fragments of 'Early Spring Burpless' cucumber.…”

Studi Penyiapan Akar Berkualitas untuk Uji Kromosom dan Penggandaan Kromosom Planlet Hasil Kultur Anter Anthurium

“…Tunas-tunas untuk studi penyiapan akar berkualitas diperoleh dengan menanam kalus hasil kultur anter Anthurium dengan bakal tunas pada medium WinartoTeixeira (WT) tanpa penambahan hormon (Winarto et al 2011a). Tujuh hingga delapan kalus dengan bakal tunas ditanam dalam botol jam yang telah diisi dengan medium semi padat dengan volume medium 35 ml per botol untuk meningkatkan keseragaman eksplan.…”

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

“…Spadices with 50% of their stigmas in a receptive condition-indicated by high secretion of a sticky substance at the tip of the stigma (see Fig. 1 in Winarto et al 2011)-were harvested from 20.1 ± 2.03-days-old flowers (counted from when the spathe started to open) between the second and fourth flower. Spadices were then sterilized by placing them under tap water for 30-60 min then immersed in a 1% pesticide solution of 50% benomyl (Benlox Ò 50 WP, Dharma Guna Wibawa Ltd, Jakarta, Indonesia) and 20% streptomycin sulphate (Agrept Ò 20WP, Mastalin Mandiri Ltd, Jakarta, Indonesia) for 30 min and rinsed with sterile distilled water 5 times (5 min each rinse).…”

“…Fast-type callus was generally green to lightgreen while the slow-type was reddish-yellow to light reddish-yellow. The fast type could produce a high number of regenerated shoots (up to 12 shoots per explant) in 4-5 months after the first subculture while the slow type took 9-12 months to produce 1-3 regenerated-shoots per explant (Rachmawati 2005;Winarto et al 2011). Callus clusters were then sliced into 27 mm 3 ''blocks'' (3 9 3 9 3 mm, All values in mg/l BAP 6-benzylaminopurine, 2,4-D 2,4-dichlorophenoxy acetic acid, Kin kinetin, NAA a-naphthalene acetic acid, TDZ thidiazuron, PGR plant growth regulator l 9 w 9 h).…”

“…This study thus concentrated on formulating a new basal medium suitable for developing a protocol of half-anther culture of Anthurium based on the potential media and selected-cultivar tested previously. In this process, which lead to the successful development to of a callus-induction medium for Anthurium halfanthers, a unique combination of micro-and macronutrients was hence assembled, and that medium was henceforth termed Winarto-Teixeira (WT) medium, already recognized by the plant science peer community (Winarto et al 2010b(Winarto et al , 2011. Reliable induction and proliferation of callus derived from half-anther culture, followed by shoot regeneration and root formation, are the main objectives to develop an anther culture protocol for Anthurium.…”