Figure 1.
Kernel phenotype and protein accumulation pattern of Illinois Protein Strains.
(A) Kernel translucency and vitreousness of IHP, IRHP, ILP and IRLP. (B) Zein and non-zein accumulation pattern of B73, IHP, IRHP, ILP and IRLP analyzed by 15% SDS-PAGE. Protein from 500 µg of maize flour was loaded in each lane. M, protein markers from top to bottom being 250, 150, 100, 75, 50, 37, 25, 20, 15 and 10 kDa. The size of each zein band is indicated with numbers in the “kDa” column.
Table 1.
Protein and amino acid composition analysis of four Illinois Protein Strains (IHP, IRHP, ILP and IRLP) and B73.
Figure 2.
Rebalancing of zein and non-zein ratio in IRLP and IHP.
Zeins and non-zeins extracted from six “non-green” and “green” kernels (lanes 1–6 and 7–12, respectively), each from IRLP x P6z1RNAi/- (A) and IHP x P6z1RNAi/- (B). They were scored for GFP with fluorescent light microscope and analyzed individually by 15% SDS-PAGE. (C) Comparison of zein and non-zein ratio in total protein between IHP/-; -/- and IHP/-; P6z1RNAi/-. (D) Comparison of zein and non-zein accumulation patterns in W64A, W64Ao2, IHP/-; -/- and IHP/-; P6z1RNAi/-. The total protein loaded in each lane was equal to 500 µg of maize flour. M, protein markers in the top panels of (A) and (B) from top to bottom being 37, 25, 20, 15 and 10 kDa, and in (D) and the bottom panels of (A) and (B) being 250, 150, 100, 75, 50, 37, 25, 20, 15 and 10 kDa. The size for each zein band is marked with numbers in the “kDa” column.
Figure 3.
Ear phenotype of P6z1RNAi backcrossed to B73, IRLP and IHP.
(A) Kernels were cut in half for observation under a natural light (upper panel) and fluorescence dissection microscope (bottom panel). Opaque phenotype in kernels from the ear with P6z1RNAi backcrossed to B73 for two generations and IRLP x P6z1RNAi/- is linked to the expression of GFP. In IHP x P6z1RNAi/-, the non-green and green kernels are both vitreous. V&N, vitreous and non-green; O&G, opaque and green; V&G, vitreous and green; The ear with P6z1RNAi backcrossed to B73 for two generations (B) and IRLP x P6z1RNAi/- (C) show 1∶1 ratio of vitreous and opaque segregation. Two vitreous and opaque kernels in each ear are indicated by white and black arrows, respectively. (D) In IHP x P6z1RNAi/-, kernels were uniformly vitreous.
Figure 4.
Ear phenotype of P6z1RNAi/- x IRLP (A) and P6z1RNAi/- x IHP (B).
Vitreous and opaque kernels segregate as 1∶1 ratio in both crosses. Two representative kernels for each phenotype are indicated by white and black arrows, respectively.
Table 2.
Protein and amino acid composition analysis of seeds with genotype IHP/-; -/-, IHP/-; P6z1RNAi/-, IRLP/-; -/-, IRLP/-; P6z1RNAi/- and W64Ao2.