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Sparks, C. A.

Publications and source records attributed to Sparks, C. A..

3 recordsLinked to original sources

Can small reductions in Rubisco content improve nitrogen use in wheat without negatively impacting biomass or grain yield?

In this study, the level of Rubisco protein was reduced in wheat using RNAi, to test the hypothesis that photosynthesis, growth, and grain yield could be maintained whilst improving nitrogen use efficiency. The RNAi Rubisco wheat plants, with a Rubisco activity of less than 70% of wild type (WT) plant levels, had reduced photosynthesis, reductions in leaf and stem biomass and decreased seed yield. Interestingly, in the wheat RNAi Rubisco lines that had a small (<30%) reduction in Rubisco activity, the seed number, total seed weight and harvest index were comparable to that of WT type plants. However, no improvement in photosynthetic nitrogen use efficiency (PNUE) was evident in any of the RNAi Rubisco lines. Notably, PNUE was lower than for WT wheat plants in the RNAi lines with more than a 30% reduction in Rubisco activity. This result was unexpected and caused by an accumulation of N in both the leaves and seeds. At present we do not have an explanation for this but one possible hypothesis is that it could be due to slower growth caused by a reduction in source strength in the RNAi plants, which in turn resulted in changes to carbon and nitrogen allocation. HighlightWheat RNAi plants with small reductions in the amount and activity of Rubisco had a similar biomass and total seed weight to that of untransformed controls but no improvement in nitrogen use efficiency was evident.

plant biology↗

A high soluble-fibre allele in wheat encodes a defective cell wall peroxidase responsible for dimerization of ferulate moieties on arabinoxylan

Increasing dietary fibre (DF) intake is an important target to improve health and an attractive strategy for this is to increase the fibre content of staple foods, particularly white bread which is the staple food in many countries. DF in wheat white flour is derived principally from the endosperm cell wall polysaccharide arabinoxylan (AX) and the water-extractable form of this (WE-AX) accounts for the majority of soluble dietary fibre (SDF), which is believed to confer particular health benefits. We previously identified QTLs for soluble dietary fibre (SDF) on 1B and 6B chromosomes in wheat in biparental populations. Here we show that the 6B high SDF allele encodes a peroxidase protein (PER1-v) with a single missense compared to the more common low SDF form (PER1). Wheat lines with the natural PER1-v allele and with an induced knock-out mutation in PER1 showed similar characteristics of reduced dimerization of ferulate associated with water-extractable WE-AX. Decreased ferulate dimerization is associated with decreased cross-linking of the WE-AX chains and increased solubility of AX. Transiently expressed PER1_RFP fusion driven by native promoter in wheat endosperm was shown to localise to cell walls whereas PER1-v_RFP did not; we therefore propose that PER1-v lacks capacity to dimerise AX ferulate in vivo due to mis-localisation. PER1 is the first peroxidase reported to be responsible for oxidative coupling of ferulate on AX, a key process in all grass cell walls. Understanding its role and the effect of variants on AX properties offers a route to control the properties of wheat DF in the human diet.

plant biology↗

Efficient CRISPR/Cas-mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties

The CRISPR/Cas technology has recently become a molecular tool of choice for gene function studies in plants as well as crop improvement. Wheat is a globally important staple crop with a well annotated genome and there is plenty of scope for improving its agriculturally important traits using genome editing technologies, such as CRISPR/Cas. As part of this study we targeted three different genes in hexaploid wheat Triticum aestivum: TaBAK1-2 in the spring cultivar Cadenza as well as Ta-eIF4E and Ta-eIF(iso)4E in winter cultivars Cezanne, Goncourt and Prevert. The primary transgenic lines carrying CRISPR/Cas-induced indels were successfully generated for all targeted genes. As winter wheat varieties are generally less amenable to genetic transformation, the successful experimental methodology for transformation and genome editing in winter wheat presented in this study will be of interest to the research community working with this crop.

plant biology↗