Fig. 5
Functional analysis of EMB976 via overexpressing truncated EMB976 CDS in WT. a Yellow leaf phenotype of T1 transgenic plants overexpressing the truncated EMB976-14M (bar = 1.5 cm). b RT-qPCR analysis of mRNA levels of the endogenous EMB976 in transgenic lines. c RT-qPCR analysis of mRNA levels of total EMB976 in transgenic lines. d RT-PCR analysis of precursor transcripts of chloroplast genes with specific primers. The number after the decimal point indicates the intron of the gene. e Splicing efficiency of clpP1 intron 2 and ycf3 intron 1 in the cosuppression lines. f Phenotype of T1 plants overexpressing truncated EMB976-14M in rdr6-11 (bar = 1.5 cm). g RT-PCR analysis of mRNA levels of endogenous EMB976, total EMB976, clpP1 and ycf3 precusors in the transformants. h RT-qPCR analysis of mRNA levels of endogenous EMB976 in the transformants. i RT-qPCR analysis of mRNA levels of total EMB976 in the transformants. j RT-qPCR analysis of splicing efficiency of plastid clpP1 intron 2, ycf3 intron 1 and ycf3 intron 2 in the transformants. For RT-qPCR, the values are means of three technique replicates (bars indicate SD). Asterisks indicate significant differences between wild type (WT) and transgenic plants (Student’s t test, *, P < 0.05 and **, P < 0.01). Two to three T1 transformants were analyzed for RT-PCR and RT-qPCR