Deletion mutagenesis identifies a haploinsufficient role for γ-Zein in opaque2 endosperm modification

Lingling Yuan, Yongchao Dou, Shahryar F. Kianian, Chi Zhang, David R. Holding

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)119-130
Number of pages12
JournalPlant physiology
Volume164
Issue number1
DOIs
StatePublished - Jan 1 2014

Fingerprint

Zein
Endosperm
zein
mutagenesis
Mutagenesis
endosperm
Zea mays
corn
Proteins
Modifier Genes
protein bodies
modifiers (genes)
RNA Sequence Analysis
Quantitative Trait Loci
proteins
quantitative trait loci
lysine
Lysine
sequence analysis
chromosomes

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

Cite this

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.

In: Plant physiology, Vol. 164, No. 1, 01.01.2014, p. 119-130.

Research output: Contribution to journalArticle

Yuan, Lingling ; Dou, Yongchao ; Kianian, Shahryar F. ; Zhang, Chi ; Holding, David R. / Deletion mutagenesis identifies a haploinsufficient role for γ-Zein in opaque2 endosperm modification. In: Plant physiology. 2014 ; Vol. 164, No. 1. pp. 119-130.
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