Effect of primary culture medium composition on high frequency somatic embryogenesis in different Coffea species

Abstract

High frequency somatic embryogenesis can be obtained over a 7-8 month culture period when using current routing coffee tree micropropagation protocols. To reduce this response time and improve the embryo formation yield, eight different media were tested for primary culture. These media differed from the classically used ones by their mineral nitrogen and plant growth regulator concentrations. An increase from 0.66 to 0.75 in the NO 3 / NO 3 +NH 4 ratio and a 2-fold lower plant growth regulator concentration in the primary culture medium led to substantial improvements in terms of rapidity and embryo/plantlet regeneration frequencies. Embryo development time was reduced by up to 3 months with a 5-fold increase in the number of formed embryos. These results were obtained for the two cultivated coffee tree species, Coffea canephora and C. arabica, and for a wild one, C. heterocalyx, but not for a second wild species, C. sp. Moloundou showing a species-specific response. The new conditions described in this paper led to a substantial enhancement that should be particularly helpful for clonal propagation and genetic engineering of cultivated coffee plants.

Plant regeneration via indirect somatic embryogenesis and optimisation of genetic transformation in Coffea arabica L. cvs. Caturra and Catuaí

“…The regeneration medium R (van Boxtel and Berthouly, 1996) contained 17.6 µM of BAP whereas the S3 medium (Quiroz-Figueroa et al 2002) contained 2.32 µM of kinetin and 0.27 µM of napthaleneacetic acid (NAA). Somatic embryo development from embryogenic callus or suspension cultures has been accomplished previously in coffee mainly on semisolid culture media (Barry- Etienne et al 2002b;Barrueto et al 2004;Samson et al 2006). Temporary immersion culture systems have improved the development of somatic embryos in Citrus deliciosa, Hevea brasiliensis, Musa spp and C. arabica F1 hybrid (Caturra x E531) (Escalant et al 1994;Cabasson et al 1997;Etienne et al 1997;Barry-Etienne et al 1999;Barry-Etienne et al 2002a).…”

EFISIENSI MEDIA KULTUR DAN APLIKASI TEMPORARY IMMERSION SYSTEM PADA EMBRIOGENESIS SOMATIK KOPI ARABIKA / Efficiency of Culture Media and Aplication Temporary Immersion Systemon on Somatic Embryogenesis Arabica Coffee

“…Pada kultur in vitro sukrosa pada umumnya diberikan dalam jumlah 2 -5% (George et al 2008). Dalam kultur in vitro kopi melalui embriogenesis somatik, sukrosa diberikan pada kisaran 2 -4% tergantung pada tahap perkembangan embrio (Etienne 2005;Samson et al 2006;Rezende et al 2012).…”

Micropropagation of Coffea sp. by somatic embryogenesis cultures technique

“…Vi nh©n gièng ® ®−îc øng dông trong nh©n nhanh cµ phª vµ nu«i cÊy ph«i v« tÝnh lµ kü thuËt tiªn tiÕn cã nhiÒu tiÒm n¨ng ph¸t triÓn nh©n gièng cµ phª quy m« lín [10,11].…”

“…Sù biÖt hãa tÕ bµo ph«i v« tÝnh cµ phª ®−îc ®iÓu khiÓn bëi m«i tr−êng vËt lý hay c¸c chÊt kÝch thÝch sinh tr−ëng vµ sù c©n b»ng auxin/cytokinin trong nu«i cÊy [5,13], dinh d−ìng kho¸ng [11]. Ph«i v« tÝnh cµ phª ® ®−îc nghiªn cøu vÒ m« häc [10] vµ ® x©y dùng ®−îc ph−¬ng ph¸p chän dßng m« sÑo mµu vµng chanh cã hiÖu suÊt ph¸t sinh ph«i v« tÝnh cao [13].…”

“…H¹t cµ phª mÊt søc nÈy mÇm sau 2 n¨m tån tr÷. Nh¸nh chåi v−ît dïng cho gi©m cµnh th× cã h¹n vµ ph−¬ng ph¸p gi©m cµnh th−êng mang theo mÇm bÖnh tõ c©y mÑ ë cµ phª [12].Vi nh©n gièng ® ®−îc øng dông trong nh©n nhanh cµ phª vµ nu«i cÊy ph«i v« tÝnh lµ kü thuËt tiªn tiÕn cã nhiÒu tiÒm n¨ng ph¸t triÓn nh©n gièng cµ phª quy m« lín [10,11].Nu«i cÊy ph¸t sinh vµ t¸i sinh ph«i v« tÝnh ë c©y cµ phª phô thuéc vµo nhiÒu ®iÒu kiÖn nh− t×nh tr¹ng sinh lý cña l¸ ®−a vµo nu«i cÊy, lo¹i m« l¸ [12], kÝch th−íc mÉu nu«i cÊy nhá hay lín, ® nu«i cÊy ®Ønh chåi v−ît, thêi gian cÊy truyÒn [14], ®iÒu kiÖn khÝ hËu, kiÓu gen [8].Sù biÖt hãa tÕ bµo ph«i v« tÝnh cµ phª ®−îc ®iÓu khiÓn bëi m«i tr−êng vËt lý hay c¸c chÊt kÝch thÝch sinh tr−ëng vµ sù c©n b»ng auxin/cytokinin trong nu«i cÊy [5,13], dinh d−ìng kho¸ng [11]. Ph«i v« tÝnh cµ phª ® ®−îc nghiªn cøu vÒ m« häc [10] vµ ® x©y dùng ®−îc ph−¬ng ph¸p chän dßng m« sÑo mµu vµng chanh cã hiÖu suÊt ph¸t sinh ph«i v« tÝnh cao [13].…”