In another study, Armin et al. [ 20] also speculated that the application of NAA completely inhibited the growth of single nodes of in vitro grown potato plantlets.
The leaves, stem and root were collected from 45 days old whole in vitro grown potato plantlets and immediately frozen in liquid nitrogen, subsequently stored at -80°C until used.
Based on the initial real-time quantitative reverse transcription PCR (qRT-PCR) assay of the in vitro cultured potato plants, the transcript levels of the StuPPO1 genes were reduced by 68 to 98% in six amiRPPO1 series transgenic lines (clones).
Relative expression level of target gene, was assayed using in vitro cultured potato leaf and stem tissues by qRT-PCR; the data is the average of three technical repeats and their standard deviation of using one original sample from each transgenic line; n.v., no value; n.t.
For cloning and sequencing of the GSL genes, genomic DNA was isolated from in vitro shoots of potato, Solanum tuberosum L., cv Iwa based on the method described by Bernatzky and Tanksley [ 46].
The successful in vitro multiplication of potatoes depends on the presence of a suitable combination of auxins with gibberellic acid (GA3) in the propagation medium [ 4– 7].
Adding exogenous GA3 with different auxins is a good way to reduce micropropagation time and increase the number of plantlets for in vitro micropropagation of potatoes [ 25].
For the collection of root, stem, old leaf and young leaf samples, in vitro raised plantlets of potato cv.
During in vitro culture, the transgenic potato plants showed a normal morphologic characteristic and growth pattern in patatin hpRNAi lines 2, 4, 7, 8 (See Additional file 2A).
