• Xuye Du

      Articles written in Journal of Genetics

    • Cloning and characterization of novel fast $\omega$-gliadin genes in Triticum monococcum

      Xuye Du Jinglan Zhao Xin Ma Huayan Yin Hongwei Wang Anfei Li Lingrang Kong

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    • Molecular cloning and functional characterization of two novel high molecular weight glutenin subunit genes in Aegilops markgrafii


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      The high molecular weight glutenin subunits (HMW-GS) in bread wheat are major determinants of the viscoelastic properties of dough and the end-use quality of wheat flour. Two novel HMW-GSs, 1Cx1.1 and 1Cy9.1, from the diploid speciesAegilops markgrafii (CC) were identified in the present study. The corresponding open-reading frames of the genes of 1Cx1.1 and 1Cy9.1 were isolated and sequenced using allele-specific polymerase chain reaction. Sequence comparison demonstrated that the HMW-GSs from Ae. markgrafii possess a similar primary structure to the homologous proteins in wheat and related species. A tandem tripeptide exists in the central repetitive domain of 1Cx1.1, and this unique structure is very rare in the HMW-GSs of other genomes. To confirm the authenticity of these isolated endogenous HMW-GS, the heterologous proteins produced by removing the signal peptides expressed by E. coli exhibited the same electrophoretic mobility as the native proteins. Subsequently, the singleprotein was purified at a sufficient scale for incorporation into flour to performsodium dodecyl sulphate (SDS) sedimentation testing. Notably, the SDS sedimentation volume was less with the addition of 1Cx1.1 than it was with 1Cy9.1.

    • Cloning and characterization of a novel low-molecular-weight glutenin subunit gene with an unusual molecular structure of Aegilops uniaristata


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      Low-molecular-weight glutenin subunits (LMW-GSs) are one of the important factors for the dough processing quality. In this study, a novel LMW-GS, designated LMW-N13, from the wheat relative species Aegilops uniaristata PI 554421 was cloned and characterized. Unlike previously published LMW-GSs, LMW-N13 has a large molecular weight and is the largest LMW-GS published thus far. Sequence alignments demonstrated that LMW-N13 is a LMW-i-type subunit but contains nine cysteine residues which is one more than typical LMW-i-type subunits. In addition, four insertions are present in the repetitive domain that resulted in the large molecular weight. In vitro analysis showed that LMW-N13 could improve the dough quality of different base flours.

    • Identification of NRAMP4 from Arabis paniculata enhance cadmium tolerance in transgenic Arabidopsis


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      Arabis paniculata has been reported as a hyperaccumulator and functions in cadmium (Cd) tolerance and accumulation. However, the genes involved in Cd stress resistance in A. paniculata are still unknown. In this work, genes of the natural resistanceassociated macrophage proteins (NRAMPs) were characterized in A. paniculata, and their evolutionary relationship and expression patterns were analysed. Expression profiles indicated that ApNRAMPs showed large differences in response to Cd stress. It was highly induced by Cd in root and shoot tissues. To investigate the function of ApNRAMP4 under Cd stress, ApNRAMP4 was cloned and expressed in yeast and Arabidopsis. The results indicated that yeast and Arabidopsis expressing ApNRAMP4 showed normal growth under Cd stress. In addition, transgenic yeast and Arabidopsis showed the ability to concentrate Cd. Under 20 μM CdCl2, Cd concentrations in wild type (WT) and transgenic yeast were 3.11 and 5.92 mg/kg, respectively. Cd concentrations in root tissues of WTand transgenic Arabidopsis were 0.18 and0.54 mg/kg, respectively. In shoot tissues of WT and transgenic Arabidopsis, Cd concentrations were 0.13 and 0.49 mg/kg, respectively. This report provides genomic information on hyperaccumulator A. paniculata. In addition, the present work identified key NRAMP genes that may serve as resources for heavy metal phytoremediation.

    • Arabis paniculata ApHIPP3 increases Cd tolerance by interacting with ApCHC1


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      Cadmium (Cd) is a common hazardous element that shows potential chronic toxicity in plants and animals. Arabis paniculata functioning as a hyperaccumulator has been found in the Yunnan–Guizhou plateau in China. This study characterizes the function of ApHIPP3 in A. paniculata to Cd stress, showing that the expression of ApHIPP3 was highly induced by Cd stress. Arabidopsis thaliana overexpressing ApHIPP3 showed stronger growth potential than wild type (WT). Cd accumulation capacity was significantly higher intransgenic A. thaliana than in WT. In addition, transgenic A. thaliana showed the ability to inhibit electrolyte leakage and eliminate reactive oxygen species (ROS). A strong interaction of ApHIPP3 with the clathrin heavy chain ApCHC1, which is known to play an important role in biotic and abiotic stresses, could be detected by a yeast two-hybrid assay. This was further confirmed by pull-down and co-immunoprecipitation (Co-IP) assays. Overall, these results demonstrate the function of ApHIPP3 to Cd stress and suggest a regulatory mechanism in response to Cd stress.

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