Transformation of tetraploid potato cv Spunta and cv. Kennebec with TALEN for site mutagenesis

MASSA GABRIELA ALEJANDRA; CECILIA DÉCIMA ONETO; NATALIA ALMASIA; COLAVITA, MÓNICA LILIANA; VALERIA PERALTA; CECILIA VAZQUEZ-ROVERE; FEINGOLD SERGIO ENRIQUE

Congreso; 11 th International plant molecular biology congress; 2015

In the last years site-directed mutagenesis paved the way to predictableand precise editing of specific DNA sequences. This technology presents enormouspotential in plant breeding, especially in clonal crops. All genome editingtechniques involve a meganuclease that cleaves DNA and machinery that guidesthe meganuclease to a specific DNA sequence. Transcription activator?like (TAL) effector proteins guide FokImeganuclases in TALEN technique.   In this study we usedacetolactate synthase gene (als) as a target for non-homologous end joining(NHEJ). Leaves from in vitro propagated shoot cultures of Solanum tuberosum cv. Spunta and cv. Kennebec were transformedusing Agrobacterium. Two different tetraploid potato cultivars (Spuntaand Kennebec) were used under their specific protocols. Transformationefficiency (transgenic events per transformed explants) was 2.66 % and 4.65 %,respectively. Four to eleven shoots per explant were obtained. A total of 8 transgenicevents from cv. Spunta and 22 from cv. Kennebec were obtained. The higherefficiency in Kennebec can be attributed to major modifications introduced tothe Millam transformation protocol, not presented here.   In the transformed plants, both cv Spunta and cv.Kennebec, the expression of TALEN was induced by β-stradiol and ALS-target was sequenced in eachtransgenic plant. At this time the sequenced regions analysis is being carried out in order to confirmthe presence of mutations in alsgene. This work is the first attempt on genome editing in two commercial potatocultivars like Spunta and Kennebec and if successful will open the door forinnovative incremental potato breeding.