Maize kernel zein accumulation of liao2345/<i>o2</i>-1, liao2345/<i>o2</i>-2, liao2345, and CA339 in mature seeds, detected by SDS/PAGE.

<p>Total zein loaded in each lane is equal to 1 mg of mature seed endosperm, and the relative molecular weights of proteins are marked at the right side, the unit is kDa.</p

Multiple alignment of partial sequences of the sixth exon of the <i>O2</i> gene.

<p>Sequences of x15544 and B73 are supplied by GenBank and MaizeSequence, respectively. The other five sequences are sequenced by us. Dhuang212 and liao2345 are wild-type inbred lines in China. Dashes indicate gaps and asterisks indicate identical residues. The stop codons are indicated by blue characters, and the insertions and deletions are indicated by red characters.</p

Analysis of chromosome segments introgression of <i>O2</i> based on SSR markers mapping to bin7.01.

<p>The positions of SSR markers are based on a map from IBM2 2008 Neighbors 7. The red fragments (A) are of recurrent parent origin, blue fragments (B) are of donor origin, and gray fragments (U) represent markers that have no polymorphism between the two parents, and have no information for that genotype.</p

Structure of the <i>O2</i> gene and specific site of each sequence primer.

<p>Structure of the <i>O2</i> gene and specific site of each sequence primer.</p

Examination of the <i>O2</i> gene transcript abundance in developing endosperm (18 DAP) by RT-PCR.

<p>O2RT1 and O2RT2 are two specific primers spanning the third and the fifth exon of the <i>O2</i> gene, the amplified region of ORT2 is larger than that of O2RT1; GAPDH is the reference gene. The templates of RT-PCR are labeled in each line, H₂O represents the blank control; RNA represents the negative control; cDNA is used to detect abundance of the <i>O2</i> transcript.</p

Maize kernel phenotypes of liao2345/<i>o2</i>-1, liao23455/<i>o2</i>-2, and liao2345; <i>o2o2</i>, <i>o2o2</i>, <i>O2O2</i>.

<p>A: Photographs of intact kernels taken under normal light on light box. B: Photographs of intact kernels taken with transmitted light. C: Photographs of decapped kernels taken under normal light.</p

Test of three SSR markers inside the <i>O2</i> gene by PAGE.

<p>phi112, umc1066, and phi057 can distinguish the three genotype of the <i>O2</i> gene. The amplified region of phi112 located in the promoter of <i>O2</i> the gene; and umc1066 in the first exon of the <i>O2</i> gene; and phi057 in the sixth exon of the <i>O2</i> gene.</p

Name and pedigree of maize lines.

<p>Name and pedigree of maize lines.</p

Multiple alignment of deduced OPAQUE2 protein sequences.

<p>Dashes indicate gaps and asterisks indicate identical residues. The SSR site of umc1066 is underlined and the SSR site of phi057 is indicated by double underlining. The basic motif is marked with a red box, and the leucine zipper domain is marked by a yellow box. Blue ellipses indicate sites that differ between the two <i>o2</i> NILs.</p

Analysis of promoter difference between two <i>o2</i> NILs by normal PCR on agarose gel.

<p>The size of the amplified region of three pairs primers are 582 bp, 648 bp, and 723 bp. This region contains the TATA box and initiation codon of <i>O2</i> gene.</p