@article{fdi:010053607,
  title = {{A}grobacterium-mediated genetic transformation of {C}offea arabica ({L}.) is greatly enhanced by using established embryogenic callus cultures},
  author = {{R}ibas, {A}lessandra and {D}echamp, {E}. and {C}hampion, {A}ntony and {B}ertrand, {B}. and {C}ombes, {M}arie-{C}hristine and {V}erdeil, {J}. {L}. and {L}apeyre, {F}. and {L}ashermes, {P}hilippe and {E}tienne, {H}.},
  editor = {},
  language = {{ENG}},
  abstract = {{B}ackground: {F}ollowing genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. {W}hen gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. {E}mbryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including {C}offea arabica. {R}esults: {W}e identified the conditions required for successful long-term proliferation of embryogenic cultures in {C}. arabica and designed a highly efficient and reliable {A}grobacterium tumefaciens-mediated transformation method based on these conditions. {T}he transformation protocol with {LBA}1119 harboring p{B}in 35{S} {GFP} was established by evaluating the effect of different parameters on transformation efficiency by {GFP} detection. {U}sing embryogenic callus cultures, co-cultivation with {LBA}1119 {OD}600 = 0.6 for five days at 20 degrees {C} enabled reproducible transformation. {T}he maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation) and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%). {A}t the histological level, successful transformation was related to the number of proembryogenic masses present. {A}ll the selected plants were proved to be transformed by {PCR} and {S}outhern blot hybridization. {C}onclusion: {M}ost progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. {T}his is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. {O}ur results make {A}grobacterium-mediated transformation of embryogenic cultures a viable and useful tool both for coffee breeding and for the functional analysis of agronomically important genes.},
  keywords = {},
  booktitle = {},
  journal = {{B}mc {P}lant {B}iology},
  volume = {11},
  numero = {},
  pages = {92},
  ISSN = {1471-2229},
  year = {2011},
  DOI = {10.1186/1471-2229-11-92},
  URL = {https://www.documentation.ird.fr/hor/fdi:010053607},
}