Multiplication of Musa from excised stem tips.
ABSTRACT Rapidly multiplying cultures were established from excised shoot tips of two dessert banana clones ('Philippine Lacatan' and 'Grande Naine') and two plantain clones ('Pelipita' and 'Saba'). Apices cultured on semi-solid media produced single shoots while apices placed in liquid media produced shoot clusters. Individual shoots were induced to form multiple shoot clusters by longitudinally splitting the shoot through the apex. Shoot multiplication was stimulated maximally by 5 mg l-1 benzylaminopurine. Rooted plants were produced by treating shoots with auxins. Growth rates based on increase in f. wt of the four clones were compared. During a 4-week culture period 'Pelipita' showed a fivefold increase in f. wt while 'Grande Naine', 'Philippine Lacatan' and 'Saba' showed increases exceeding tenfold.
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ABSTRACT: RESUMO. O objetivo deste trabalho foi estabelecer um protocolo para a multiplicação in vitro de Bastão-do-imperador (Etlingera elatior). Brotos axilares foram inoculados em meio MS contendo as seguintes concentrações de reguladores de crescimento durante a fase de isolamento: testemunha; BAP 2,25 mg L -1 ; BAP 2,25 mg L -1 + ANA 0,93 mgL -1 ; BAP 2,25 mg L -1 + AIA 0,87 mg L -1 ; BAP 4,95 mg L -1 ; BAP 4,95 mg L -1 + ANA 0,93 mg L -1 ; BAP 4,95 mg L -1 + AIA 0,87 mg L -1 . As brotações provenientes desta fase foram subcultivadas em um novo meio de cultura MS: testemunha; BAP 2,25 mg L -1 ; BAP 3,37 mg L -1 ; BAP 4,50 mg L -1 ; BAP 2,25 mg L -1 + ANA 1,12 mg L -1 ; BAP 3,37 mg L -1 + ANA 2,25 mg L -1 ; BAP 4,50 mg L -1 + ANA 3,37 mg L -1 e avaliou-se a taxa de multiplicação dos brotos por repetição. Durante a fase de isolamento, o melhor tratamento foi BAP 4,95 mg L -1 + AIA 0,87 mg L -1 . Durante a fase de multiplicação, ocorreram em média três a quatro brotações por repetição, independentemente das concentrações e combinações de reguladores de crescimento utilizadas. As plântulas enraizaram e formaram touceiras no meio MS sem reguladores de crescimento, resultando em oito a dez plântulas por touceira. ABSTRACT. Establishing a protocol for in vitro multiplication of philippine wax flower (Etlingera elatior) Jack RM Sm. The objective of the present paper was to establish a protocol for in vitro multiplication of the Philippine wax flower. Axillary shoots were inoculated in MS medium, holding the following concentrations of plant growth regulators during the isolation stage: without plant growth regulator; benzylaminopurine(BAP) 2.25 mg L -1 ; BAP 2.25 mg L -1 + naphthaleneacetic acid (NAA) 0.93 mg L -1 ; BAP 2.25 mg L -1 + indoleacetic acid (IAA) 0.87 mg L -1 ; BAP 4.95 mg L -1 ; BAP 4.95 mg L -1 + NAA 0.93 mg L -1 ; BAP 4.95 mg L -1 + IAA 0.87 mg L -1 and shootings from this stage were cultivated in a new MS medium following different combinations of plant growth regulators: without plant growth regulator; BAP 2.25 mg L -1 ; BAP 3.37 mg L -1 ; BAP 4.50 mg L -1 ; BAP 2.25 mg L -1 + NAA 1.12 mg L -1 ; BAP 3.37 mg L -1 + NAA 2.25 mg L -1 ; BAP 4.50 mg L -1 + NAA 3.37 mg L -1 and shooting multiplication rate per replicate was evaluated. The best treatment, during the isolation stage, was BAP 4.95 mg L -1 + IAA 0.87 mg L -1 . During the multiplication stage, there were three to four shootings per replicate, regardless of the concentrations and combinations of plant growth regulators used. The seedlings rooted, forming clusters on MS medium without plants growth regulators, resulting in eight to ten seedlings per cluster.Acta Scientiarum Agronomy 12/2010; 32(4). DOI:10.4025/actasciagron.v32i4.4830 · 0.63 Impact Factor
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ABSTRACT: Regenerated plants were established using male inflorescence of Musa acuminata cv. Berangan. Explants were cultured on Murashige and Skoog (MS) solid medium supplemented with three different concentrations of 6-benzylaminopurine (BAP). After 2 weeks, whitish bud-like structures (WBLS) were obtained and MS media supplemented with 70.0 µM BAP was observed as the best media for the growth of WBLS. After 3 months, shoot-like structures emerged and MS media supplemented with 31.0 µM BAP gave a large number of shoot formation. Normal looking plantlets were obtained within 4 -6 months with an average of 80 -130 shoots regenerated from each male inflorescence. Random amplified polymorphic DNA (RAPD) was carried out to determine the clonal fidelity on in vitro Musa acuminata cv. Berangan micropropagated from male inflorescence derived from the same mother plant. Eighteen arbitrary decamer primers were used to amplify DNA from in vitro plants materials. All RAPD profiles from regenerated plants were monomorphic thus no somaclonal variation was detected. A total of 81 bands were scored from PCR amplification of genomic DNA from 15 micropropagated plants. This was further confirmed by the value of similarity index which was equaled to 1. This result implied that male inflorescence could be used as an alternative explant for commercial planting Keywords. in vitro, clonal propagation, male inflorescence, banana, similarity index, RAPD IINTRODUCTION In vitro propagation has played a key role in clonal propagation for obtaining a large number of homogenous regenerated plants and breeding of plaintains and bananas (Musa spp.) (Pierik, 1987). The application of micropropagation for clonal propagation of Musa spp. have been reported by several authors using different explant sources such as meristems (Ma and Shii 1972; 1974; Banerjee et. al., 1986), shoot tips (Cronauer and Krikorian, 1984; Wong, 1986; Vuylsteke, 1998; Kanchanapoom and Chanadang, 2001) and floral apices (Cronauer and Krikorian, 1985a; 1985b; Balakrishnamurthy and Sree Rangaswamy, 1988; Doreswamy and Sahijram, 1989). These methods were successfully performed on a wide range of banana cultivars. RAPD technique has successfully been used for the assessment of clonal fidelity in chestnut (Carvalho et al., 2003), Piper spp. (Chaveerach et al., 2002) and turmeric (Neeta et al., 2001). The validation of RAPD bands obtained is crucial through repetitions of experiments performed (Innis et al., 1998). RAPD is also applicable for studies on diversity in Ocimum spp. (Singh et al., 2004), Heliconia spp. (Prakash et al., 1998), gingers (Rout et al., 1998) and Musa spp. (Bhat and Jarret, 1995, Howell et al., 1994, Kaemmer et al., 1992). In this study, the method established for in vitro propagation from male inflorescence offers an efficient and relatively simple method for clonal and mass propagation of Musa spp. compared to previous reports. Moreover, no studies on the assessment of clonal fidelity on the male inflorescence derived regenerants were carried out. Based on this study, male inflorescence could be used as an alternative explant instead of meristems cultures currently used for commercial propagation. In addition, male inflorescence displayed low risk of latent contamination which is a major problem in banana micropropagation.
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ABSTRACT: Banana shoot tips extracted from in vitro grown plant materials were encapsulated in different types of alginate beads. The results obtained after conservation for one month at 4˚C indicated that single layered synseeds (where the encap-sulated shoot tip was covered by one layer of the artificial endosperm) containing activated charcoal were the best con-struction in stimulating conversion of synseeds, where they expressed the highest; conversion frequency, number of shoots/synseed, length of shoot and fresh weight/shoot cluster. These parameters decreased when the shoot tips were encapsulated within two layers of endosperm, except the ex vitro conversion. Conversion of banana synseeds was in-fluenced by explant orientation inside the alginate beads, the highest conversion frequency was obtained when the shoot tip was directed upward inside the synthetic endosperm. Synthesis of seed coats around the artificial endosperm im-proved the conversion frequency, number of shoots/synseed and shoot length.