Molecular characterization of LMW-GS genes from C, N, U and S&lt;sup&gt;s&lt;/sup&gt; genomes among &lt;i&gt;Aegilops&lt;/i&gt; species

Wang, Siyu; Chen, D.; Guo, Guangfang; Zhang, T.; Jiang, Shanshan; Shen, Xing; Perović, Dragan; Prodanovic, S.; Yan, Yueming

doi:10.1556/crc.2012.0006

Cited by 7 publications

(8 citation statements)

References 34 publications

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“…Genomic DNA was extracted from dry seeds by use of CTAB protocol (Murray and Thompson 1980). The w-secalin complete gene sequences (accessions FJ561478, FJ816038, FJ823443, X60294, AF000227, and FJ561465) from GenBank (www.ncbi.nlm.nih.gov/) were used for designing allelic specific PCR (AS-PCR) primers according to Wang et al (2012a) named as forward Sec1 (5 0 -ATGAAGACCTTACTCATGCTTG-3 0 ) and reverse Sec2 (5 0 -CTATACCACTACTA CAAATGGT-3 0 ). The AS-PCR amplifications were performed in a total volume of 50 mL containing 2.5 U LA Taq polymerase (TaKaRa), 60 ng of template DNA, 25 mL 2X GC buffer I (MgCl 2 plus), 0.4 mM dNTP, 0.5 mM of each primer, and double-distilled H 2 O.…”

Section: Dna Extraction As-pcr Amplification and Cloning Of Secalinmentioning

confidence: 99%

Fast identification of wheat 1BL.1RS translocation by reversed-phase ultra-performance liquid chromatography (RP-UPLC)

et al. 2013

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Abstract. The 1BL.1RS chromosomal translocation in wheat is the result of replacement of the short arm of chromosome 1B of wheat by the short arm of chromosome 1R of rye, which had been widely used as a parental line in worldwide wheat breeding, resulting in a high percentage of wheat cultivars containing this translocation. A fast and reliable approach to identify this translocation is highly desirable in modern wheat breeding. This study compared reversed-phase ultraperformance liquid chromatography (RP-UPLC), acidic polyacrylamide gel electrophoresis (A-PAGE), liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), allelic-specific PCR, and reversed-phase highperformance liquid chromatography (RP-HPLC) approaches to identify the 1BL.1RS translocation in 76 bread wheat cultivars. Two gliadin bands in the Gli-B1 region of A-PAGE separation were confirmed by LC-MS/MS to be omega secalins from the 1BL.1RS translocation, and they can be used as reliable protein markers for identifying the translocation. A few specific minor peaks eluted at 12-13 min on the RP-UPLC patterns can readily differentiate the 1BL.1RS translocation. Of the 76 wheat cultivars tested, 40 were identified as carrying the 1BL.1RS translocation by RP-UPLC, which was consistent with the results of A-PAGE, HPLC, and PCR. Compared with other established methods, RP-UPLC showed a clear advantage in fast identification of the 1BL.1RS translocation with higher reliability and lower costs, and it is therefore ideal for large-scale screening of the 1BL.1RS translocation in wheat breeding.

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Section: Dna Extraction As-pcr Amplification and Cloning Of Secalinmentioning

confidence: 99%

Fast identification of wheat 1BL.1RS translocation by reversed-phase ultra-performance liquid chromatography (RP-UPLC)

et al. 2013

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“…Previous reports indicated that the U and C genomes are closely related to each other . The N‐terminal sequences of LMW‐GS from the A genome were similar to those from the U genome, suggesting an evolutionary relationship . The correlations between allelic variations of Glu‐3 and breadmaking quality in common wheat are well established .…”

Section: Introductionmentioning

confidence: 99%

“…Subsequently, some novel LMW‐GS genes were isolated and characterized from Ae. umbellulata , but their functional properties and application potential are still unclear …”

Section: Introductionmentioning

confidence: 99%

“…26 The N-terminal sequences of LMW-GS from the A genome were similar to those from the U genome, suggesting an evolutionary relationship. 27 The correlations between allelic variations of Glu-3 and breadmaking quality in common wheat are well established. 28 Recent studies have shown that Glu-A3a and Glu-B3h contribute to superior flour quality.…”

mentioning

confidence: 99%

“…umbellulata, but their functional properties and application potential are still unclear. [25][26][27] Recently, we developed the derived CNU609 line by crossing between Chinese Spring (CS, T. aestivum L., 2n = 6x = 42, AABBDD) and the wheat-Ae. umbellulata (2n = 2x = 14, UU) 1U(1B) substitution line.…”

mentioning

confidence: 99%

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Low‐molecular‐weight glutenin subunits from the 1U genome of Aegilops umbellulata confer superior dough rheological properties and improve breadmaking quality of bread wheat

Wang

Zhen

et al. 2017

J Sci Food Agric

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Molecular characterisation of high molecular weight glutenin allele Glu-B1h encoding 1Bx14+1By15 subunits in bread wheat (Triticum aestivum L.)

et al. 2014

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In this study, the authentic high molecular weight glutenin (HMW-GS) allele Glu-B1 h encoding for subunits 1Bx14 and 1By15 from German bread wheat cultivars Hanno and Imbros was identified and cross-verified by a suite of established protein analysis technologies, including sodium dodecyl sulfate-polyacrylamide gel electrophoresis, reversed-phase high-performance liquid chromatography, reversed-phase ultra-performance liquid chromatography, and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). The complete encoding sequences were isolated by allele-specific PCR, and consist of 2367 bp for 1Bx14 and 2151 bp for 1By15 and encode 789 and 717 amino acid residues, respectively. The deduced molecular masses of two subunit genes were 82 340.13 Da and 74 736.13 Da, corresponding well to those determined by MALDI-TOF-MS. The presence and authenticity of 1Bx14 and 1By15 subunits were further confirmed by liquid chromatography coupled to tandem mass spectrometry and heterologous expression in E. coli. Comparative analysis demonstrated that 1Bx14 possessed one deletion and 20 single-nucleotide polymorphism variations compared with seven other Glu-B1 x-type HMW-GS genes that mainly resulted from C–T substitutions, whereas compared with five other Glu-B1 y-type HMW-GS genes, 1By15 displayed few variations. Phylogenetic analysis based on the complete coding sequences of the published HMW-GS genes showed that 1Bx14 had a high divergence with other 1Bx subunit genes, whereas 1By15 displayed greater similarity with 1By20. A possible evolutionary route for 1Bx14 gene formation is proposed, which might have resulted from an intra-strand illegitimate recombination event that occurred ~1.32 million years ago.

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Molecular characterization of LMW-GS genes from C, N, U and S<sup>s</sup> genomes among <i>Aegilops</i> species

Cited by 7 publications

References 34 publications

Fast identification of wheat 1BL.1RS translocation by reversed-phase ultra-performance liquid chromatography (RP-UPLC)

Fast identification of wheat 1BL.1RS translocation by reversed-phase ultra-performance liquid chromatography (RP-UPLC)

Low‐molecular‐weight glutenin subunits from the 1U genome of Aegilops umbellulata confer superior dough rheological properties and improve breadmaking quality of bread wheat

Molecular characterisation of high molecular weight glutenin allele Glu-B1h encoding 1Bx14+1By15 subunits in bread wheat (Triticum aestivum L.)

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