In non-legume the symbiotic-like responses were observed only in nim1/npr1 mutants, or in NahG transformants, both of which suppress the SA signaling (Fig. 1).
The co-bombarded plasmid was detectable in ~30 50% of the paromomycin-resistant transformants regardless of which co-bombarded plasmid was used.
Second, we constructed a yeast open reading frame (ORF) galactose-induced overexpression array by generating a genome-wide set of yeast transformants, each of which carries an individual plasmid-born and sequence-verified ORF derived from the Saccharomyces cerevisiae full-length EXpression-ready (FLEX) collection.
Interactions were assessed by plating several double transformants from "non-selective" media (media that allows for identification of the double transformants) on which growth is possible only if there exists an interaction between the test subjects.
Thus, we did not screen for mutant strains with drastic defects of photosynthesis, but our screening procedure had to differentiate between oxygen-producing transformants and transformants which become anaerobic in the light.
It was used in the present studies to generate the three different transformants populations each of which incorporated one of gene segments of interest, tat (150 bp), rev (243 bp) and nef1 (642 bp).
Figure 2a shows that 10 transformants were arbitrarily chosen for PCR analysis, of which transformants 2, 5, 6, and 10 (lanes 2, 5, 6, and 10, respectively) did not exhibit the expected 1.1 kb band.
A total of 29 independent transformants were examined, out of which 5 lines showed strong growth defects and sterility following treatment with dexamethasone.
To minimize positional effect of reporter gene expression, at least 1000 transformants on the membrane were collected to create a mixed population of transformants, which were cultured and assayed for the strength of GFP florescence.
Consequently, 21 independent transformant lines were obtained, of which 12 lines represented single copy lines, as confirmed by Southern blot analysis using bar and sGFP gene probes (Figure 1B, Table S1).
