TY - JOUR
T1 - molecular cloning and characterization of six novel HMW-GS genes from aegilops speltoides and aegilops kotschyi
AU - Ma, Chaoying
AU - Yang, Yang
AU - Li, Xiaohui
AU - Ge, Pei
AU - Guo, Guangfang
AU - Subburaj, Saminathan
AU - Zeller, Friedrich J.
AU - Hsam, Sai L.K.
AU - Yan, Yueming
PY - 2013/6
Y1 - 2013/6
N2 - Six novel high molecular weight glutenin subunits (HMW-GS) from Aegilops speltoides (SS, 2n = 2x = 14) and Aegilops kotschyi (UUSS, 2n = 4x = 28) were identified by SDS-PAGE and designated as ASy15*, AKx1*, AKx3*, AKx2.3, AKy20* and AKy8*, respectively. Their complete open reading frames (ORFs) were cloned and sequenced by allele-specific PCR (AS-PCR). Sequence comparison demonstrated that these novel genes displayed high single-nucleotide polymorphisms (SNP) and InDel variations. In particular, AKy8* had an extra cysteine residue at position 140 in the central repetitive domain, while AKx2.3 had an unusually long repetitive domain (816 aa), which might have positive effects on gluten quality. Phylogenetic analysis showed that AKx1* and AKx3* belonged to the 1S genome, AKx2.3 to the 1U genome, and AKy20* and AKy8* most likely to the 1S genome. The divergence between the 6 HMW-GS genes from the two Aegilops species and those from Triticum species occurred 4.77-23.54 MYA. The authenticity of these isolated endogenous HMW-GS genes was confirmed through heterologous expression in Escherichia coli and Western blotting.
AB - Six novel high molecular weight glutenin subunits (HMW-GS) from Aegilops speltoides (SS, 2n = 2x = 14) and Aegilops kotschyi (UUSS, 2n = 4x = 28) were identified by SDS-PAGE and designated as ASy15*, AKx1*, AKx3*, AKx2.3, AKy20* and AKy8*, respectively. Their complete open reading frames (ORFs) were cloned and sequenced by allele-specific PCR (AS-PCR). Sequence comparison demonstrated that these novel genes displayed high single-nucleotide polymorphisms (SNP) and InDel variations. In particular, AKy8* had an extra cysteine residue at position 140 in the central repetitive domain, while AKx2.3 had an unusually long repetitive domain (816 aa), which might have positive effects on gluten quality. Phylogenetic analysis showed that AKx1* and AKx3* belonged to the 1S genome, AKx2.3 to the 1U genome, and AKy20* and AKy8* most likely to the 1S genome. The divergence between the 6 HMW-GS genes from the two Aegilops species and those from Triticum species occurred 4.77-23.54 MYA. The authenticity of these isolated endogenous HMW-GS genes was confirmed through heterologous expression in Escherichia coli and Western blotting.
KW - Aegilops HMW-GS
KW - Heterologous expression
KW - Molecular cloning
UR - http://www.scopus.com/inward/record.url?scp=84878996070&partnerID=8YFLogxK
U2 - 10.1111/pbr.12046
DO - 10.1111/pbr.12046
M3 - Article
AN - SCOPUS:84878996070
SN - 0179-9541
VL - 132
SP - 284
EP - 289
JO - Plant Breeding
JF - Plant Breeding
IS - 3
ER -