An efficient protocol for large-scale plantlet production from male floral meristems of Musa spp. cultivars Virupakshi and Sirumalai

Abstract

In vitro propagation has played a key role for obtaining large numbers of virus free, homogenous plants, and for breeding of plantains and bananas (Musa spp.). Explant sources utilized for banana micropropagation include suckers, shoot tips, and floral buds. The present study employed male floral meristems as explant material for micropropagation of hill banana ecotypes (AAB) 'Virupakshi' and 'Sirumalai.' Immature male floral buds were collected from healthy plants from hill banana growing areas. Exposure of explants to ethyl alcohol (70%, v/v) for 30 s, then mercuric chloride (0.1%, w/v) for 30 s, followed by three independent rinses of 5 min each in autoclaved, double-distilled water satisfactorily reduced the contamination. Male floral bud explants were cultured on Murashige and Skoog (MS) basal medium supplemented with different combinations of 6-benzylaminopurine (BAP), coconut water, naphthaleneacetic acid, gibberellic acid, and additional supplements. MS medium supplemented with 5 mg l −1 BAP and coconut water (15%) was the most efficient media for shoot initiation and multiple shoot formation (15 shoots from a single part of a floral bud). The best response for shoot elongation was obtained using the combination of basal MS, 5 mg l −1 BAP, 1 mg l −1 naphthaleneacetic acid and 1.5 mg l −1 gibberellic acid. Regenerated shoots were rooted in basal MS medium within 15-20 d. The rooted plantlets were transferred to a soil mixture and maintained at a temperature of 25±2°C for 10 d and then at room temperature (30-32°C) for 2 wk, before transferring to a greenhouse. The regenerated plantlets showed 100% survival.

Effect of Medium and Cytokinin Types on Banana Micropropagation during Multiplication Stage

“…The above mentioned results are in agreement with those of Gübbük and Pekmezci (2004), Gubbuk and Pekmezci (2006), Kalimuthu et al (2007), Shiragi et al (2008), Al-Amin et al (2009), Bhosale et al (2011), Mahadev et al (2011, Jafari et al (2013), Rahman et al (2013), Anbazhagan et al (2014), Mahdi et al (2014), Ngomuo and Ndakidemi (2014) and Shankar et al (2014). All of them found that BAP and kinetin increased the shoot number initiated from shoot tips of banana during micropropagation through tissue culture.…”

Penggunaan Aksis Jantung Pisang untuk Penyediaan Sumber Eksplan Bebas Bakteri

“…Induksi tunas in vitro dari jantung pisang telah dilaporkan di mancanegara, yaitu menggunakan eksplan berupa aksis, sisir (hands) berukuran sekitar 0,5 cm, dan seperempat bagian jantung yang dipotong secara vertikal dengan panjang 0,5−2 cm (Darvari et al, 2010;Khalil et al, 2002;Krikorian et al, 1993Krikorian et al, , 1999Mahadev et al, 2011;Resmi dan Nair, 2007). Pada kultur in vitro pisang, sitokinin berupa benzyladenine (BA) paling sering digunakan untuk multiplikasi tunas.…”

“…Mahadev et al (2011) menggunakan ZPT yang lebih bervariasi, yaitu BA, naphthalene acetic acid (NAA), asam giberelin (GA 3 ), ditambah dengan air kelapa (mengandung difenil urea) untuk menginduksi tunas in vitro dari eksplan irisan jantung.…”

“…Darvari et al (2010) melakukan subkultur sebanyak dua kali untuk meregenerasikan eksplan pisang Barangan yang berupa sisir. Mahadev et al (2011) bahkan melakukan subkultur berulang atau secara frekuentif setiap satu bulan dan melakukan empat tahap regenerasi, yaitu induksi tunas, elongasi tunas, multiplikasi tunas, dan induksi perakaran dengan menggunakan media yang berbeda-beda dalam meregenerasi irisan jantung.…”

“…Secara keseluruhan, faktor multiplikasi tunas yang dihasilkan pada penelitian ini lebih tinggi dibanding dengan yang telah dilakukan oleh Darvari et al (2010) dan Mahadev et al (2011). Darvari et al (2010) dilaporkan mampu menghasilkan 13,5 tunas/ sisir, sedangkan Mahadev et al (2011) mampu menghasilkan 60 tunas/jantung.…”