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Biomedical subjects

Michael J Holdsworth

Publications and source records attributed to Michael J Holdsworth.

3 recordsLinked to original sources

Transgenesis has less impact on the transcriptome of wheat grain than conventional breeding.

Detailed global gene expression profiles have been obtained for a series of transgenic and conventionally bred wheat lines expressing additional genes encoding HMW (high molecular weight) subunits of glutenin, a group of endosperm-specific seed storage proteins known to determine dough strength and therefore bread-making quality. Differences in endosperm and leaf transcriptome profiles between untransformed and derived transgenic lines were consistently extremely small, when analysing plants containing either transgenes only, or also marker genes. Differences observed in gene expression in the endosperm between conventionally bred material were much larger in comparison to differences between transgenic and untransformed lines exhibiting the same complements of gluten subunits. These results suggest that the presence of the transgenes did not significantly alter gene expression and that, at this level of investigation, transgenic plants could be considered substantially equivalent to untransformed parental lines.

Breeding↗

A transcriptomics resource for wheat functional genomics.

Grain development, germination and plant development under abiotic stresses are areas of biology that are of considerable interest to the cereal community. Within the Investigating Gene Function programme we have produced the resources required to investigate alterations in the transcriptome of hexaploid wheat during these developmental processes. We have single pass sequenced the cDNAs of between 700 and 1300 randomly picked clones from each of 35 cDNA libraries representing highly specific stages of grain and plant development. Annotated sequencing results have been stored in a publicly accessible, online database at http://www.cerealsdb.uk.net. Each of the tissue stages used has also been photographed in detail, resulting in a collection of high-quality micrograph images detailing wheat grain development. These images have been collated and annotated in order to produce a web site focused on wheat development (http://www.wheatbp.net/). We have also produced high-density microarrays of a publicly available wheat unigene set based on the 35 cDNA libraries and have completed a number of microarray experiments which validate their quality.

Journal Article↗

Transcripts of Vp-1 homeologues are misspliced in modern wheat and ancestral species.

The maize (Zea mays) Viviparous 1 (Vp1) transcription factor has been shown previously to be a major regulator of seed development, simultaneously activating embryo maturation and repressing germination. Hexaploid bread wheat (Triticum aestivum) caryopses are characterized by relatively weak embryo dormancy and are susceptible to preharvest sprouting (PHS), a phenomenon that is phenotypically similar to the maize vp1 mutation. Analysis of Vp-1 transcript structure in wheat embryos during grain development showed that each homeologue produces cytoplasmic mRNAs of different sizes. The majority of transcripts are spliced incorrectly, contain insertions of intron sequences or deletions of coding region, and do not have the capacity to encode full-length proteins. Several VP-1-related lower molecular weight protein species were present in wheat embryo nuclei. Embryos of a closely related tetraploid species (Triticum turgidum) and ancestral diploids also contained misspliced Vp-1 transcripts that were structurally similar or identical to those found in modern hexaploid wheat, which suggests that compromised structure and expression of Vp-1 transcripts in modern wheat are inherited from ancestral species. Developing embryos from transgenic wheat grains expressing the Avena fatua Vp1 gene showed enhanced responsiveness to applied abscisic acid compared with the control. In addition, ripening ears of transgenic plants were less susceptible to PHS. Our results suggest that missplicing of wheat Vp-1 genes contributes to susceptibility to PHS in modern hexaploid wheat varieties and identifies a possible route to increase resistance to this environmentally triggered disorder.

Alternative Splicing↗