Muhammad NAWAZ, İhsan ULLAH, Naeem IQBAL, Muhammad Zafar IQBAL, Muhammad Aslam JAVED

Improving in vitro leaf disk regeneration system of sugarcane (Saccharum officinarum L.) with concurrent shoot/root induction from somatic embryos

Genome engineering experiments are impeded by poor performance of regeneration systems. The present study was aimed at establishing a short and cost-effective in vitro regeneration system for elite sugarcane cultivars through simultaneous shoot/root induction. The innermost spindle leaf and shoot tip were used as explants. For callus induction, Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) was used and 5.0 mg/L of 2,4-D supported maximum callus induction (84.5%). Three-week-old calli were treated with different levels of benzylaminopurine (BAP) ranging from 0.00 to 3.5 mg/L in MS medium, where 2.5 mg/L BAP was proven to be the best level for regeneration. In a multiplication and root formation medium, 0.5 mg/L naphthalene acetic acid supported the maximum number of roots per plant. Finally, a direct somatic embryogenesis protocol was established, competent enough for simultaneous root/shoot induction. The results indicated that the plantlets were established within 12 weeks only. This in vitro regeneration protocol was fast and cost-effective and may be used for large-scale in vitro regeneration of sugarcane cultivars to save time and resources. The sugarcane cultivar SPF-234 remained the most responsive, followed by HSF-242 and CPF-246.

Anahtar Kelimeler:

Key words: Sugarcane, somatic embryogenesis, fast regeneration

Improving in vitro leaf disk regeneration system of sugarcane (Saccharum officinarum L.) with concurrent shoot/root induction from somatic embryos

Keywords:

Key words: Sugarcane, somatic embryogenesis, fast regeneration,

PDF

___

Ahloowalia BS, Maretzki A (1983). Plant regeneration via somatic embryogenesis in sugarcane. Plant Cell Rep 2: 21–25.
Ahmadabadi M, Rauf S, Bock R (2007). A leaf-based regeneration and transformation system for maize (Zea mays L.). Transgenic Res 16: 437–448.
Ali K, Afghan S (2001). Rapid multiplication of sugarcane through micro propagation technique. Pak Sugar Journal 16: 11–14.
Arencibia AD, Carmona R, Cornide MT, Menedez E, Molina P (2000). Transgenic sugarcane (Saccharum species). In: Bajaj YPS, editor. Biotechnology in Agriculture and Forestry 46. Transgenic Crops 1. Heidelberg, Germany: Springer, pp. 188– 20
Baksha R, Alam R, Karim MZ, Paul BSK, Hossain MA, Miah MAS, Rahman ABMM (2002). In vitro shoot tip culture of sugarcane (Saccharum officinarum) variety LSD28. Biotechnol 1: 67–72.
Bano S, Jabeen M, Rahim F, Ilahi I (2005). Callus induction and regeneration in seed explants of rice (Oryza sativa cv. Swat-II). Pak J Bot 37: 829–836.
Block MD, Herrera EL, Vanmontagu M, Schell J, Zambryski P (1984). Expression of foreign genes in regenerated plants and in their progeny. EMBOJ 3: 1681–1690.
Brisibe EA, Miyake H, Taniguchi T, Maeda E (1994). Regulation of somatic embryogenesis in long term callus cultures of sugarcane (Saccharum officinarum L.). New Phytol 126: 301– 30
Chattha MA, Abida A, Muhammad I, Akhtar A (2001). Micropropagation of sugarcane (Saccharum species hybrid). Pak Sugar Journal 16: 2–6.
Chengalrayan K, Gallo-Meagher M (2011). Effect of various growth regulators on shoot regeneration of sugarcane. In Vitro Cell Dev-Pl 7: 434–439.
Desai NS, Suprasanna P, Bapat VA (2004). Simple and reproducible protocol for direct somatic embryogenesis from cultured immature inflorescence segments of sugarcane (Saccharum spp.). Curr Sci 87: 764–768.
Eldessoky DS, Ismail RM, Hadi AA, Hadi A, Abdallah N (2011). Establishment of regeneration and transformation system of sugarcane cultivar GT54-9 (C9). GM Crops 2: 1–9.
Enriquez GA, Trujillo LE, Menendez C, Vazquez RI, Tiel K, Dafhnis F, Arrieta J, Selman G, Hernandez L, Arencibia AD (2000). Sugarcane (Saccharum hybrid) genetic transformation mediated by Agrobacterium tunefaciens: production of transgenic plants expressing proteins with agronomic and industrial value. In: Plant Genetic Engineering: Towards the Third Millennium: Proceedings of the International Symposium on Plant Genetic Engineering, Havana, Cuba, 6–10 December 1999; pp. 76–80.
Finer JJ, Kriebel HB, Becwar MR (1989). Initiation of embryonic callus and suspension cultures of eastern white pine (Pinus strobes L.). Plant Cell Rep 8: 203–206.
Geetha S, Padmanabhan D (2001). Effect of hormones on direct somatic embryogenesis in sugarcane. Sugar Technol 3: 120– 1
Hiei Y, Ohta S, Komari T, Kumashiro T (1994). Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6: 271–282.
Jadhav AB, Vaidya ER, Aher VB, Pawar AM (2001). In vitro multiplication of co–86032 sugarcane (S. officinarum) hybrid. Indian J Agr Sci 71: 113–115.
Jones HD, Doherty A, Wu H (2005). Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat. Plant Methods 1: 5–6.
Khalil SM (2002). Regeneration via somatic embryogenesis and microprojectile-mediated co-transformation of sugarcane. Arab J Biotechnol 5: 19–32.
Khan SA, Rashid H, Chaudhary FM, Chaudhary Z, Afroz A (2008). Rapid micro propagation of three elite Sugarcane (Saccharum officinarum L.) varieties by shoot tip culture. Afr J Biotechnol 7: 2174–2180.
Kharinarain RP, Dolgikh IY, Guzhov LY (1996). Selection of media for mass regeneration of sugarcane plants from callus culture. Plant Physiol 43: 97–100.
Klóska L, Cegielska-Taras T, Piętka T (2012). Regeneration capacity of selected genotypes of white mustard (Sinapis alba L.). In Vitro Cell Dev-Pl 48: 180–188.
Kumar S, Bhat V (2012). High-frequency direct plant regeneration via multiple shoot induction in the apomictic forage grass Cenchrus ciliaris L. In Vitro Cell Dev-Pl 48: 241–248.
Lal N (2003). High frequency plant regeneration from sugarcane callus. Sugar Technol 5: 89–91.
Malabadi RB, Mulgund GS, Nattaraja K, Kumar SV (2011). Induction of somatic embryogenesis in different varieties of sugarcane (Saccharam officinarum L.). RPB 1: 39–48.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Plant Physiol 15: 473–497.
Nasir ID, Jahangir GZ, Qamar Z, Rahman Z, Hussain T (2011). Maintaining the regeneration potential of sugarcane callus for longer span. Afr J Agric Res 6: 113–119.
Naz S (2003). Micro propagation of promising varieties of sugarcane and their acclimatization response. In: Activities on Sugar Crops in Pakistan. Proceedings of Fourth Workshop on Research Development, pp. 1–9.
Nishimura A, Aichi I, Matsuoka M (2007). A protocol for Agrobacterium- mediated transformation in rice. Nat Protoc 1: 2796–2802.
Okumuş V, Pirinç V, Onay A, Başaran D (2011). In vitro propagation of Diyarbakır watermelons and comparison of direct-seeded and transplanted watermelon. Turk J Biol 35: 601–610.
Popelka JC, Altpeter F (2003). Evaluation of rye (Secale cereale L.) inbred lines and their crosses for tissue culture response and stable genetic transformation of homozygous rye inbred line L22 by biolistic gene transfer. Theor Appl Genet 107: 583–590. Rashid H, Ghani RA, Chaudhry Z, Naqvi SMS, Quraishi A (2002). Effect of media, growth regulators and genotypes on callus induction and regeneration in wheat (Triticum aestivum). J Biotechnol 1: 49–54.
Razi-ud-din, Shah SS, Hussan SW, Ali S, Zamir R (2004). Micropropagation of sugarcane though bud culture. Sarhad J Agr 20: 1–
Snyman SJ, Meyer GM, Koch AC, Banasiak M, Watt M (2011). Applications of in vitro culture systems for commercial sugarcane production and improvement. In Vitro Cell Dev-Pl 47: 234–249.
Verma SK, Yucesan BB, Gurel S, Gurel E (2011). Indirect somatic embryogenesis and shoot organogenesis from cotyledonary leaf segments of Digitalis lamarckii Ivan., an endemic medicinal species. Turk J Biol 35: 743–750.
Xu LP, Que YX, Xu JS, Fang SR, Zhang MQ, Chen YQ, Chen RK (2008). Establishment of genetic transformation system and obtaining transgenic sugarcane (var.badila) transformed with RS gene. Sugar Technol 10: 128–132.