Modified expression of ZmMYB167 in Brachypodium distachyon and Zea mays leads to increased cell wall lignin and phenolic content

One of the challenges to enable targeted modification of lignocellulosic biomass from grasses for improved biofuel and biochemical production lies within our limited understanding of the transcriptional control of secondary cell wall biosynthesis. Here, we investigated the role of the maize MYB transcription factor ZmMYB167 in secondary cell wall biosynthesis and how modified ZmMYB167 expression in two distinct grass model species affects plant biomass and growth phenotypes. Heterologous expression of ZmMYB167 in the C3 model system Brachypodium led to mild dwarf phenotypes, increased lignin (~7% to 13%) and S-lignin monomer (~11% to 16%) content, elevated concentrations of cell wall-bound p-coumaric acid (~15% to 24%) and reduced biomass sugar release (~20%) compared to controls. Overexpression of ZmMYB167 in the C4 model system Zea mays increased lignin (~4% to 13%), p-coumaric acid (~8% to 52%) and ferulic acid (~13% to 38%) content but did not affect plant growth and development nor biomass recalcitrance. Taken together, modifying ZmMYB167 expression represents a target to alter lignin and phenolic content in grasses. The ZmMYB167 expression-induced discrepancies in plant phenotypic and biomass properties between the two grass model systems highlight the challenges and opportunities for MYB transcription factor-based genetic engineering approaches of grass biomass

Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.)

Genetic transformation of perennial ryegrass (Lolium perenne L.) is critical for fundamental and translational research in this important grass species. It often relies on Agrobacterium-mediated transformation of callus tissue. However, callus induction is restricted to a few genotypes that respond well to tissue culture. Here, we report callus induction from different perennial ryegrass genotypes and explants, such as shoot tips, seeds, and anthers, which were transformed with several plasmids for functional genomics. β-glucuronidase (GUS) histochemical staining showed the LmdsRNAbp promoter sequence was active in stigmas, spikelets, anthers, and leaves. We also transformed calli with plasmids allowing gene silencing and gene knock-out using RNA interference and CRISPR/Cas9, respectively, for which genotypic and phenotypic investigations are ongoing. Using 19 different constructs, 262 transgenic events were regenerated. Moreover, the protocol regenerated a doubled haploid transgenic event from anther-derived calli. This work provides a proof-of-concept method for expanding the range of genotypes amenable to transformation, thus, serving research and breeding initiatives to improve this important grass crop for forage and recreation

Prospectus, October 14, 1974

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Exploring the experiences of healthcare professionals in South Africa and Uganda around communicating with children about life-threatening conditions: a workshop-based qualitative study to inform the adaptation of communication frameworks for use in these settings

OBJECTIVES: This study aimed to explore how published communication frameworks could be amended to ensure applicability and cultural appropriateness for professionals to support family-centred conversations by investigating’ healthcare professionals’ (1) experiences of providing support to families when a caregiver or a dependent child (<18 years old) has a life-threatening condition, (2) perceived challenges for caregivers and healthcare professionals in communicating with children about illness, (3) perceptions of how clinicians could be equipped to facilitate conversations between caregivers and children about an adult or the child’s own life-threatening condition and (4) suggestions for amendments to previously published guidelines to ensure cultural relevance in South Africa and Uganda. DESIGN: A qualitative study involving two 2-day workshops with embedded focus group discussions, break out rooms and consensus discussions. SETTING: Health and social care and third sector organisations in South Africa and Uganda. PARTICIPANTS: Thirty-two professionals providing care to families affected by life-threatening conditions in South Africa or Uganda who were aged 18 years or older and able to converse in English. RESULTS: Participants identified obstacles to having conversations with caregivers about children and to telling children about serious illness during consultations. These included patients’ beliefs about illness, medicine and death, language barriers between families and the healthcare team, and emotional and practical challenges for professionals in having these conversations. Culturally appropriate adaptations were made to previously published communication frameworks for professionals to support family-centred conversations. CONCLUSIONS: Culturally sensitive communication frameworks could help healthcare professionals to talk with families about what children need to know when they or a caregiver have a serious illness. More broadly, effective communication could be facilitated by promoting healthcare professionals’ and communities’ understanding of the benefits of telling children about illness within the family. Together these strategies may mitigate the psychological impact of global disease on children and their families

Modification of perceived beer bitterness intensity, character and temporal profile by hop aroma extract

The effect of hop aroma on perceived bitterness intensity, character and temporal profile of beer was investigated. A hop aroma extract was added at 3 levels (0, 245, 490 mg/L) to beers at low, medium and high bitterness. Beers were evaluated for perceived bitterness intensity, harshness, roundedness and linger by a trained panel using a rank-rating technique at each bitterness level, with and without nose clips. The use of nose clips enabled the olfactory aspect to be decoupled from taste and mouthfeel aspects of bitterness perception. Results showed significant modification of perceived bitterness in beer by hop aroma depending on the inherent level of bitter-ness. These modifications were mainly driven by olfaction – in an example of taste-aroma interactions, as well as certain tactile sensations elicited by the hop aroma extract in the oral cavity. At low bitterness, beers with hop aroma added were perceived as more bitter, and of ‘rounded’ bitterness character relative to those without hop aroma. When judges used nose clips, this effect was completely eliminated but the sample was perceived to have a ‘harsh’ bitterness character. Conversely, at high bitterness, even when nose clips were used, judges still perceived beers containing hop aroma to be more bitter. These increases in bitterness perception with nose clips indicates the stimulating of other receptors, e.g. trigeminal receptors by hop aroma extract, which in tandem with the high bitterness, cause perceptual interactions enhancing bitterness intensity and also affecting bitterness character. Bitterness character attributes such as ‘round’ and ‘harsh’ were found to significantly depend on bitterness and aroma levels, with the second level of aroma addition (245 mg/L) giving a ‘rounded’ bitterness in low bitterness beers but ‘harsh’ bitterness in high bitterness beers. The impact of aroma on temporal bitterness was also confirmed with time-intensity measurements, and found to be mostly significant at the highest level of hop aroma addition (490 mg/L) in low bitterness beers. These findings represent a significant step forward in terms of understanding bitterness flavour perception and the wider impact of hop compounds on sensory perception