Development and characterization of wheat–Aegilops kotschyi 1Uk(1A) substitution line with positive dough quality parameters
Exploring novel high molecular weight glutenin subunits (HMW-GSs) from wild related species is a strategy to improve wheat processing quality. The objective of the present investigation was to identify the chromosomes of the wheat-alien introgression line N124, derived from the hybridization between Triticum aestivum with Aegilops kotschyi, and characterize the effects on quality-related traits. Fluorescence in situ hybridization karyotypes showed that N124 is a disomic 1Uk(1A) substitution line. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase high-performance liquid chromatography verified N124 expressed two HMW-GSs of the Ae. kotschyi parent. PacBio RNA sequencing and phylogenetic analysis confirmed that the two HMW-GSs were Ukx and Uky. Compared to the wheat parent, the substitution line had no obvious agronomic defects except fewer grains per spike but improved several major quality parameters. It can be served as a donor or bridge material for wheat quality improvement.
The serine proteases of Mycobacteria tuberculosis (Mtb) are important contributors to the process of bacterial invasion and its pathogenesis. In the present study, we systematically characterized the role of the Rv1043c protein in Mycobacterium infection by purifying the Rv1043c protein in Escherichia coli and constructing a Mycobacterium smegmatis (Msg) strain overexpressing Rv1043c (Msg_Rv1043c). We found that Rv1043c had serine protease activity and localized to the surface of Mtb. We determined that the optimal pH and temperature for the Rv1043c serine protease were 9.0 and 45°C, respectively. Moreover, the serine protease activity of Rv1043c was enhanced by divalent metal ions of Ca2+ and Mg2+. Site-directed mutagenesis studies demonstrated that the serine 279 residue in Rv1043c plays a catalytic role. Additionally, mouse model studies confirmed that Rv1043c significantly enhanced the survival of Msg in vivo, induced pulmonary injury and lung cell apoptosis, and promoted the release of pro-inflammatory cytokines interleukin-1β and interleukin-6 in mice. This study presents novel insights into the relationship between mycobacterial serine protease and the pathogenesis of the disease.