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Structural and expressional analysis of the B-hordein genes in Tibetan hull-less barley

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Abstract

The B-hordein gene family was analyzed from two Tibetan hull-less barley cultivars Z09 and Z26 (Hordeum vulgare subsp. vulgare). Fourteen B-hordein genes, designated BZ09-2 to BZ09-5 (from Z09) and BZ26-1 to BZ26-10 (from Z26), were sequenced. Seven of them, similar to a previously reported BZ09-1 from Z09, were predicted to encode putative active proteins each with a signal peptide, a repetitive domain, and a C-terminal region; seven of them were predicted to be pseudogenes. The B-hordein gene family was analyzed using all known representatives of B-hordein sequences and two most similar LMW-GSs of Triticum aestivum. Alignment of these seven putative proteins with known B-hordeins and two most similar LMW-GSs of T. aestivum revealed that they shared a common motif. A large variation was observed between numbers of repeat units of predicted B-hordeins of Z26 and Z09. Phylogenetic analysis revealed that all BZ26 clones were clustered in a subgroup, and BZ09-1 formed another subgroup by itself in the putative eight active genes. In addition, six 5′-upstream regulatory sequences of the B-hordein genes were isolated from the two accessions by a single oligonucleotide nested PCR, and several different mutations were identified in the cis-acting element GLM and two distinctive sequences (Z09P-2 and Z26P-3). Phylogenetic analysis of 5′-upstream regulatory regions of the B-hordein genes showed that members from the same accession were clustered together except for two distinct members. Quantitative real time PCR analysis indicated distinct expression levels of B-hordein genes in four developing stages of developing grains in two accessions. These findings suggest B-hordein genes have intrinsic differences between accessions, and this knowledge will be useful for incorporating the B-hordeins protein in barley breeding programs.

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Acknowledgments

This work was financially supported by the Special Basic Research Funds of the Ministry of Science and Technology of China (2006FY110700) and the National Science and Technology Supporting Programs (2006BAD13B02-13). The authors are thankful for the cooperation between USDA-ARS Dale Bumpers National Rice Research Center and Chengdu Institute of Biology, The Chinese Academy of Sciences. For proofreading the authors are thankful to Ellen McWhirter.

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Authors and Affiliations

  1. College of Life Sciences, Northwest A&F University, 712100, Yangling, Shaanxi, People’s Republic of China

    Zhaoxue Han

  2. College of Life Sciences, Nanjing University, 210093, Nanjing, People’s Republic of China

    Zhaoxue Han

  3. Chengdu Institute of Biology, The Chinese Academy of Sciences, 610041, Chengdu, People’s Republic of China

    Fang Wu, Guangbing Deng, Gang Qian & Maoqun Yu

  4. USDA-ARS Dale Bumpers National Rice Research Center, Stuttgart, AR, 72160, USA

    Yulin Jia

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  1. Zhaoxue Han
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  2. Fang Wu
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Correspondence to Maoqun Yu or Yulin Jia.

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Han, Z., Wu, F., Deng, G. et al. Structural and expressional analysis of the B-hordein genes in Tibetan hull-less barley. Genetica 138, 227–239 (2010). https://doi.org/10.1007/s10709-009-9415-6

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