TaMSH7: A cereal mismatch repair gene that affects fertility in transgenic barley (Hordeum vulgare L.)

Lloyd, A. B.; Milligan, Andrew S.; Langridge, Peter; Able, Jason A.

doi:10.1186/1471-2229-7-67

Cited by 47 publications

(44 citation statements)

References 39 publications

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“…Thus, in comparison to model plants (Arabidopsis and rice), few MGs have been functionally characterized in wheat. Characterised wheat MGs include TaASY1 [ 54,55 ], TaMSH7 [ 56 ], TaRAD51 [ 57,58 ], TaDMC1 [ 57,58 ], TaPSH1 [ 59 ], TaZIP4 [ 31-33 ] and RecQ-7 [ 60 ]. Given that, the assessment of whether the three modules contain known MGs was undertaken using orthology informed approaches.…”

Section: Resultsmentioning

confidence: 99%

A co-expression network in hexaploid wheat reveals mostly balanced expression and lack of significant gene loss of homeologous meiotic genes upon polyploidization

Alabdullah

Borrill

Martín

et al. 2019

Preprint

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14Polyploidization has played an important role in plant evolution. However, upon polyploidization the 15 process of meiosis must adapt to ensure the proper segregation of increased numbers of chromosomes 16 to produce balanced gametes. It has been suggested that meiotic gene (MG) duplicates return to a 17 single copy following whole genome duplication to stabilise the polyploid genome. Therefore, upon 18 the polyploidization of wheat, a hexaploid species with three related (homeologous) genomes, the 19 stabilization process may have involved rapid changes in content and expression of MGs on 20 homeologous chromosomes (homeologs). To examine this hypothesis, sets of candidate MGs were 21 identified in wheat using co-expression network analysis and orthology informed approaches. In total, 22 130 RNA-Seq samples from a range of tissues including wheat meiotic anthers were used to define 23 co-expressed modules of genes. Three modules were significantly correlated with meiotic tissue 24 samples but not with other tissue types. These modules were enriched for GO terms related to cell 25 cycle, DNA replication and chromatin modification, and contained orthologs of known MGs. Overall 26 74.4 % of genes within these meiosis-related modules had three homeologous copies which was 27 similar to other tissue-related modules. Amongst wheat MGs identified by orthology, rather than co-28 expression, the majority (93.7 %) were either retained in hexaploid wheat at the same number of 29 copies (78.4 %) or increased in copy number (15.3%) compared to ancestral wheat species. 30 Furthermore, genes within meiosis-related modules showed more balanced expression levels between 31 homeologs than genes in non-meiosis-related modules. Taken together our results do not support 32 extensive gene loss nor changes in homeolog expression of MGs upon wheat polyploidization. The 33 construction of the MG co-expression network allowed identification of hub genes and provided key 34 targets for future studies. 35 36 Author summary 37 All flowering plants have undergone a polyploidization event(s) during their evolutionary history. One 38of the biggest challenges faced by a newly-formed polyploid is meiosis, an essential event for sexual 39 reproduction and fertility. This process must adapt to discriminate between multiple related 40 chromosomes and to ensure their proper segregation to produce fertile gametes. The meiotic 41 mechanisms responsible for the stabilisation of the extant polyploids remain poorly understood except 42 in wheat, where there is now a better understanding of these processes. It has been proposed that 43 meiotic adaptation in established polyploids could involve meiotic gene loss following the event of 44 polyploidization. To test this hypothesis in hexaploid wheat, we have computationally predicted sets 45 of hexaploid wheat meiotic genes based on expression data from different tissue types, including 46 meiotic anther tissue, and orthology informed approaches. We have calculated homeolog expression 47 patterns and numbe...

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Section: Resultsmentioning

confidence: 99%

A co-expression network in hexaploid wheat reveals mostly balanced expression and lack of significant gene loss of homeologous meiotic genes upon polyploidization

Alabdullah

Borrill

Martín

et al. 2019

Preprint

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“…[30]. Autoradiography films were developed using an AGFA CP1000 Developer (AGFA, Nunawading, VIC, Australia).…”

Section: Methodsmentioning

confidence: 99%

The isolation and characterisation of the wheat molecular ZIPper I homologue, Ta ZYP1

Khoo

Able

2012

BMC Res Notes

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BackgroundThe synaptonemal complex (SC) is a proteinaceous tripartite structure used to hold homologous chromosomes together during the early stages of meiosis. The yeast ZIP1 and its homologues in other species have previously been characterised as the transverse filament protein of the synaptonemal complex. Proper installation of ZYP1 along chromosomes has been shown to be dependent on the axial element-associated protein, ASY1 in Arabidopsis.ResultsHere we report the isolation of the wheat (Triticum aestivum) ZYP1 (TaZYP1) and its expression profile (during and post-meiosis) in wild-type, the ph1b deletion mutant as well as in Taasy1 RNAi knock-down mutants. TaZYP1 has a putative DNA-binding S/TPXX motif in its C-terminal region and we provide evidence that TaZYP1 interacts non-preferentially with both single- and double-stranded DNA in vitro. 3-dimensional dual immunofluorescence localisation assays conducted with an antibody raised against TaZYP1 show that TaZYP1 interacts with chromatin during meiosis but does not co-localise to regions of chromatin where TaASY1 is present. The TaZYP1 signal lengthens into regions of chromatin where TaASY1 has been removed in wild-type but this appears delayed in the ph1b mutant. The localisation profile of TaZYP1 in four Taasy1 knock-down mutants is similar to wild-type but TaZYP1 signal intensity appears weaker and more diffused.ConclusionsIn contrast to previous studies performed on plant species where ZYP1 signal is sandwiched by ASY1 signal located on both axial elements of the SC, data from the 3-dimensional dual immunofluorescence localisation assays conducted in this study show that TaZYP1 signal only lengthens into regions of chromatin after TaASY1 signal is being unloaded. However, the observation that TaZYP1 loading appears delayed in both the ph1b and Taasy1 mutants suggests that TaASY1 may still be essential for TaZYP1 to play a role in SC formation during meiosis. These data further suggest that the temporal installation of ZYP1 onto pairing homologous chromosomes in wheat is different to that of other plant species and highlights the need to study this synaptonemal complex protein on a species to species basis.

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“…AtMutSa and AtMutSb heterodimers show a similar recognition pattern as their eukaryotic counterparts, whereas AtMutSc preferentially binds some base-base mispairs [12,13]. Apart from mismatch correction, MutSc also plays an important role during meiosis [14]. Other information about the function of plant MMR proteins has been obtained by monitoring microsatellite instability or recombination events of msh2, mlh1, or pms1 deficient plants (see [15] for a recent review).…”

Section: Introductionmentioning

confidence: 98%

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

2010

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Biochemical and immunological information concerning DNA mismatch repair proteins from higher plants is currently limited, probably due to their low abundance in vivo. An initial analysis of AtMSH2 gene expression by quantitative real-time RT-PCR indicates that calli and seedlings contain 96.7 and 1.4 cDNA copies per ng RNA, respectively, confirming that this gene is predominantly expressed in rapidly dividing tissues. In order to obtain large quantities of AtMSH2, the protein was efficiently expressed in an Escherichia coli system. The expressed gene product has an in-frame N-terminal Trx-His(6)-S-tag. The fusion protein represents about 11% of the soluble protein from IPTG-induced E. coli cells. After a two-step purification procedure the final yield accounts for 0.7 mg/g cells. Digestion of this electrophoretically homogeneous recombinant protein with enterokinase results in an intact protein with only one extra amino acid introduced at the N-terminal end. Purified intact protein was used to induce polyclonal antibodies in rabbits. These antibodies cross-react with a 110-kDa protein from cauliflower inflorescences. Together, our data describe the transcript level, cloning, expression, purification, and polyclonal antibody preparation of AtMSH2. This work will surely be useful for carrying out plant mismatch repair assays in vitro and analyzing protein expression after the exposure of plants to various stresses.

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TaMSH7: A cereal mismatch repair gene that affects fertility in transgenic barley (Hordeum vulgare L.)

Cited by 47 publications

References 39 publications

A co-expression network in hexaploid wheat reveals mostly balanced expression and lack of significant gene loss of homeologous meiotic genes upon polyploidization

A co-expression network in hexaploid wheat reveals mostly balanced expression and lack of significant gene loss of homeologous meiotic genes upon polyploidization

The isolation and characterisation of the wheat molecular ZIPper I homologue, Ta ZYP1

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

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