Based on the cytosine deaminase assay in the presence of inducing materials, wounding, sucrose, abscisic acid (ABA), and methyl jasmonate, Tr349#3 was selected. Seeds of Tr349#3 were treated with mutagen and 66,000 plants were plated on selective medi...
Based on the cytosine deaminase assay in the presence of inducing materials, wounding, sucrose, abscisic acid (ABA), and methyl jasmonate, Tr349#3 was selected. Seeds of Tr349#3 were treated with mutagen and 66,000 plants were plated on selective media containing both fluorocytosine and methyl jasmonate to get wound minus mutant(wnd^(-)) plants. 18 plants survived the second round of screening. Each of these plants was scored for the expression of the wound-inducible cytosine deaminase activity. Four of these plants had lost the wound-inducible pin2-codA expression and they were termed as wnd1, wnd4, wnd6, and wnd7. None of the selected wnd plants could be induced by wounding, methyl jasmonate, sucrose, and ABA. Because exogeneously added methyl jasmonate does not induce the pin2-codA construct, we conclude that the defect in each of the four mutations must occur downstream from methyl jasmonate production. These acquired wound minus mutant plants will be used to find out some factor(s) affecting wound minus expression of pin2-codA in Arabidopsis.