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The recessive  opaque-2  mutant gene ( o2 ) reduces α-zeins accumulation in maize endosperm, changes the amino acid composition of maize kernels, induces an opaque endosperm, and increases the lysine content of kernels. The quality protein maize (QPM) inbred line CA339 ( o2o2 ) and an elite normal inbred line liao2345 ( O2O2 ) were used to construct  o2  near-isogenic lines (NILs) by marker-assisted selection (MAS) using the co-dominant SSR marker phi057. Two specific  o2  NILs were constructed, named liao2345/ o2 -1 and liao2345/ o2 -2. However, the kernel phenotypes of the two  o2  NILs were different from each other. liao2345/ o2 -1 had the wild-type vitreous endosperm, which is similar to its recurrent parent liao2345, while the endosperm of liao2345/ o2 -2 was opaque, identical to typical  o2  mutant individuals. In comparison to their recurrent parent liao2345, the lysine concentration of liao2345/ o2 -1 was similar and the lysine concentration in liao2345/ o2 -2 was doubled. SDS-PAGE analysis indicated that liao2345/ o2 -1 had the same zeins ratio as liao2345, whereas the zeins concentration of liao2345/ o2 -2 was markedly lower. Sequence and transcript abundance analyses indicated that the CDS of two  o2  NILs are derived from CA339, but they have different promoters. The  O2  transcript of liao2345/ o2 -2 is largely inhibited because of an  rbg  transposable element inserted between the TATA box and initiator codon of liao2345/ o2 -2. We concluded that different crossing-over patterns during the process of  o2  NIL construction resulted in the different kernel phenotypes of the two  o2  NILs. We surmise that the reversion of liao2345/ o2 -1 to wild type was due to the recombination with the wild type liao2345 promoter during introgression and backcrossing. The  o2  mutant gene of donor (CA339) is a null mutant because of low  O2  expression. However, its CDS probably encodes a protein with normal function which can maintain the normal accumulation of zeins in maize endosperm.

Transposable Element rbg Induces the Differential Expression of opaque-2 Mutant Gene in Two Maize o2 NILs Derived from the Same Inbred Line