02799naa a2200457 a 450000100080000000500110000800800410001910000170006024501470007726000090022452014210023365000300165465000240168465000140170865000140172265000300173665000130176665000210177965000250180065300230182565300230184865300240187165300310189570000190192670000190194570000190196470000170198370000240200070000200202470000270204470000200207170000160209170000170210770000230212470000230214770000260217070000250219670000210222170000230224277300760226520754812018-01-15       2017    bl uuuu     u00u1 u    #d1 aBASSO, M. F.  aImproved genetic transformation of sugarcane (Saccharum spp.) embryogenic callus mediated by Agrobacterium tumefaciens.h[electronic resource]  c2017  aSugarcane (Saccharum spp.) is a monocotyledonous semi-perennial C4 grass of the Poaceae family. Its capacity to accumulate high content of sucrose and biomass makes it one of the most important crops for sugar and biofuel production. Conventional methods of sugarcane breeding have shown several limitations due to its complex polyploid and aneuploid genome. However, improvement by biotechnological engineering is currently the most promising alternative to introduce economically important traits. In this work, we present an improved protocol for Agrobacterium tumefaciens-mediated transformation of commercial sugarcane hybrids using immature top stalk-derived embryogenic callus cultures. The callus cultures are transformed with preconditioned A. tumefaciens carrying a binary vector that encodes expression cassettes for a gene of interest and the bialaphos resistance gene (bar confers resistance to glufosinate-ammonium herbicide). This protocol has been used to successfully transform a commercial sugarcane cultivar, SP80-3280, highlighting: (i) reduced recalcitrance and oxidation; (ii) high yield of embryogenic callus; (iii) improved selection; and (iv) shoot regeneration and rooting of the transformed plants. Altogether, these improvements generated a transformation efficiency of 2.2%. This protocol provides a reliable tool for a routine procedure for sugarcane improvement by genetic engineering.  aAgrobacterium radiobacter  aGenetic engineering  aSaccharum  aSugarcane  aAgrobacterium Tumefaciens  aBiomassa  aCana de açúcar  aEngenharia genética  aAgrotransformation  aCultivar SP80-3280  aImmature top stalks  aTransgenic bioenergy crops1 aDIAS, B. B. A.1 aRIBEIRO, A. P.1 aMARTINS, P. K.1 aSOUZA, W. R.1 aOLIVEIRA, N. G. DE.1 aNAKAYAMA, T. J.1 aCASARI, R. A das C. N.1 aSANTIAGO, T. R.1 aVINECKY, F.1 aJUNGMANN, L.1 aSOUSA, C. A. F. de1 aOLIVEIRA, P. A. de1 aSOUZA, S. A. C. D. de1 aCANCADO, G. M. de A.1 aKOBAYASHI, A. K.1 aMOLINARI, H. B. C.  tCurrent Protocols in Plant Biologygv. 2, n. 3, p. 221-239, Sept. 2017.