Molecular cloning and characterization of six novel HMW‐GS genes from Aegilops speltoides and Aegilops kotschyi |
| |
Authors: | Chaoying Ma Yang Yang Xiaohui Li Pei Ge Guangfang Guo Saminathan Subburaj Friedrich J. Zeller Sai L. K. Hsam Yueming Yan |
| |
Affiliation: | 1. College of Life Sciences, Capital Normal University, , Beijing, 100048 China;2. Lehrstuhl für Pflanzenzüchtung, Technische Universit?t München, , D‐85354 Freising‐Weihenstephan, Germany |
| |
Abstract: | 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. |
| |
Keywords: | molecular cloning heterologous expression Aegilops HMW‐GS |
|
|