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Deletion mutagenesis identifies a haploinsufficient role for γ-zein in opaque2 endosperm modification.
Yuan, Lingling; Dou, Yongchao; Kianian, Shahryar F; Zhang, Chi; Holding, David R.
Affiliation
  • Yuan L; Center for Plant Science Innovation, Beadle Center for Biotechnology , University of Nebraska, Lincoln, Nebraska 68588.
Plant Physiol ; 164(1): 119-30, 2014 Jan.
Article in En | MEDLINE | ID: mdl-24214534
Quality Protein Maize (QPM) is a hard kernel variant of the high-lysine mutant opaque2. Using γ-irradiation, we created opaque QPM variants to identify opaque2 modifier genes and to investigate deletion mutagenesis combined with Illumina sequencing as a maize (Zea mays) functional genomics tool. A K0326Y QPM deletion mutant was null for the 27- and 50-kD γ-zeins and abolished vitreous endosperm formation. Illumina exon and RNA sequencing revealed a 1.2-megabase pair deletion encompassing the 27- and 50-kD γ-zein genes on chromosome 7 and a deletion of at least 232 kb on chromosome 9. Protein body number was reduced by over 90%, while protein body size is similar to the wild type. Kernels hemizygous for the γ-zein deletion had intermediate 27- and 50-kD γ-zein levels and were semivitreous, indicating haploinsufficiency of these gene products in opaque2 endosperm modification. The γ-zein deletion further increased lysine in QPM in its homozygous and hemizygous states. This work identifies 27-kD γ-zein as an opaque2 modifier gene within the largest QPM quantitative trait locus and may suggest the 50-kD γ-zein also contributes to this quantitative trait locus. It further demonstrates that genome-wide deletions in nonreference maize lines can be identified through a combination of assembly of Illumina reads against the B73 genome and integration of RNA sequencing data.
Subject(s)

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Zein / Zea mays / Endosperm / Haploinsufficiency Type of study: Prognostic_studies Language: En Journal: Plant Physiol Year: 2014 Type: Article

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Zein / Zea mays / Endosperm / Haploinsufficiency Type of study: Prognostic_studies Language: En Journal: Plant Physiol Year: 2014 Type: Article