Heterologous expression and dough mixing studies of a novel cysteine-rich avenin-like protein

Chen, P.; Li, R.; Zhou, Runtao; He, Guoqing; Shewry, Peter R.

doi:10.1556/crc.38.2010.3.11

Cited by 16 publications

(21 citation statements)

References 40 publications

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“…From these results, it could be concluded that overexpression of Avenin-like b proteins generally led to improve dough elasticity, mixing tolerance and dough resistance to extension. These results were also consistent with our previous report that the direct addition of Avenin-like b proteins into flour resulted in statistically significant increases in peak resistance and mixing tolerance in an in vitro assay [29], indicating that Avenin-like b proteins had advantageous effects on mixing properties of wheat flour.…”

Section: Discussionsupporting

confidence: 92%

“…The first 18 amino acid residues corresponded to the signal peptide and the mature protein with 266 amino acid residues had a predicted molecular mass of 30 kDa [24]. The phylogenetic relationships of Avenin-like b proteins were analyzed in our previous study [29]. Immature scutella from the wheat cultivar Emai 12 were used as targets for transformation by particle bombardment.…”

Section: Resultsmentioning

confidence: 99%

“…This expression pattern was similar to the expression pattern of gliadin and glutenin genes in developing wheat seeds. In a subsequent study, incorporation of purified Avenin-like b proteins, produced in vitro , into wheat flour revealed a positive correlation between Avenin-like b proteins and dough strength by 2 g Mixograph tests [29]. In the present study, two transgenic wheat lines overexpressing avenin-like b gene were generated by particle bombardment to study the influence of Avenin-like b proteins on the functional properties of wheat flour.…”

Section: Introductionmentioning

confidence: 82%

“…The primers (forward 5′-CAGTTGGAAACCACATGT-3′, reverse 5′-TAGCACGCACCACCAGT-3′) were designed to amplify only the avenin-like b gene coding region (without the signal peptide) from the recombinant vector pLRPT-avel. Construction of the expression vector pET-32a-avel, expression and purification of the recombinant Avenin-like b proteins were carried out following the method described by Chen et al [29]. Polyclonal antibodies against Avenin-like b proteins were generated by immunizing Japanese white rabbit with the purified and renatured Avenin-like b proteins in a heterologous system.…”

Section: Methodsmentioning

confidence: 99%

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Overexpression of Avenin-Like b Proteins in Bread Wheat (Triticum aestivum L.) Improves Dough Mixing Properties by Their Incorporation into Glutenin Polymers

et al. 2013

PLoS ONE

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Avenin-like b proteins are a small family of wheat storage proteins, each containing 18 or 19 cysteine residues. The role of these proteins, with high numbers of cysteine residues, in determining the functional properties of wheat flour is unclear. In the present study, two transgenic lines of the bread wheat overexpressing avenin-like b gene were generated to investigate the effects of Avenin-like b proteins on dough mixing properties. Sodium dodecyl sulfate sedimentation (SDSS) test and Mixograph analysis of these lines demonstrated that overexpression of Avenin-like b proteins in both transgenic wheat lines significantly increased SDSS volume and improved dough elasticity, mixing tolerance and resistance to extension. These changes were associated with the increased proportion of polymeric proteins due to the incorporation of overexpressed Avenin-like b proteins into the glutenin polymers. The results of this study were critical to confirm the hypothesis that Avenin-like b proteins could be integrated into glutenin polymers by inter-chain disulphide bonds, which could help understand the mechanism behind strengthening wheat dough strength.

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Section: Discussionsupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 82%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Overexpression of Avenin-Like b Proteins in Bread Wheat (Triticum aestivum L.) Improves Dough Mixing Properties by Their Incorporation into Glutenin Polymers

et al. 2013

PLoS ONE

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“…The a-type ALPs were similar to LMW gliadins and were disengaged from the gluten polymer through S-S bonds (Clarke et al 2003;de Gregorio et al 2009). The b-type ALPs with 18/19 cysteine residues were of particular interest in determining the functional properties of gluten (Kan et al 2006), which was shown to enhance dough mixing properties when b-type ALPs were incorporated into gluten proteins (Chen et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization and evolutionary origins of farinin genes in Brachypodium distachyon L.

Subburaj

Luo

et al. 2015

J Appl Genetics

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Farinins are one of the oldest members of the gluten family in wheat and Aegilops species, and they influence dough properties. Here, we performed the first detailed molecular genetic study on farinin genes in Brachypodium distachyon L., the model species for Triticum aestivum. A total of 51 b-type farinin genes were cloned and characterized, including 27 functional and 24 non-functional pseudogenes from 14 different B. distachyon accessions. All genes were highly similar to those previously reported from wheat and Aegilops species. The identification of deduced amino acid sequences showed that b-type farinins across Triticeae genomes could be classified as b1-, b2-, b3-, and b4-type farinins; however, B. distachyon had only b3- and b4-type farinins. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) revealed that farinin genes are transcribed into mRNA in B. distachyon at much lower levels than in Triticeae, despite the presence of cis-acting elements in promoter regions. Phylogenetic analysis suggested that Brachypodium farinins may have closer relationships with common wheat and further confirmed four different types of b-type farinins in Triticeae and Brachypodium genomes, corresponding to b1, b2, b3 (group 1), and b4 (group 2). A putative evolutionary origin model of farinin genes in Brachypodium, Triticum, and the related species suggests that all b-type farinins diverged from their common ancestor ~3.2 million years ago (MYA). The b3 and b4 types could be considered older in the farinin family. The results explain the loss of b1- and b2-type farinin alleles in Brachypodium.

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Overview of the Wheat Genetic Transformation and Breeding Status in China

Han

Chang

et al. 2017

Methods in Molecular Biology

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In the past two decades, Chinese scientists have achieved significant progress on three aspects of wheat genetic transformation. First, the wheat transformation platform has been established and optimized to improve the transformation efficiency, shorten the time required from starting of transformation procedure to the fertile transgenic wheat plants obtained as well as to overcome the problem of genotype-dependent for wheat genetic transformation in wide range of wheat elite varieties. Second, with the help of many emerging techniques such as CRISPR/cas9 function of over 100 wheat genes has been investigated. Finally, modern technology has been combined with the traditional breeding technique such as crossing to accelerate the application of wheat transformation. Overall, the wheat end-use quality and the characteristics of wheat stress tolerance have been improved by wheat genetic engineering technique. So far, wheat transgenic lines integrated with quality-improved genes and stress tolerant genes have been on the way of Production Test stage in the field. The debates and the future studies on wheat transformation have been discussed, and the brief summary of Chinese wheat breeding research history has also been provided in this review.

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Heterologous expression and dough mixing studies of a novel cysteine-rich avenin-like protein

Cited by 16 publications

References 40 publications

Overexpression of Avenin-Like b Proteins in Bread Wheat (Triticum aestivum L.) Improves Dough Mixing Properties by Their Incorporation into Glutenin Polymers

Overexpression of Avenin-Like b Proteins in Bread Wheat (Triticum aestivum L.) Improves Dough Mixing Properties by Their Incorporation into Glutenin Polymers

Molecular characterization and evolutionary origins of farinin genes in Brachypodium distachyon L.

Overview of the Wheat Genetic Transformation and Breeding Status in China

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