The aim of this research was to determine the effect of development and UV‐B on flavonols and the regulation of gene activity in Vitis vinifera L. var. Sauvignon blanc grapes. Particular emphasis was ...placed on gene activity associated with the low and high fluence UV‐B responses. Flavonols, particularly quercetin and kaempferol glycosides, increased substantially upon fruit exposure due to UV‐B, with spatial analysis locating the changes to the berry skin. Of five VvFLS genes in grapes, two (VvFLS4 and 5) were found to be transcriptionally active, with VvFLS4 also being responsive to UV‐B but VvFLS5 was not. Of the transcription factors known to regulate FLS (VvMYB12, VvMYCA1 and VvWDRs), only VvMYB12 was found to be responsive to UV‐B. A number of candidate genes associated with the low and high UV‐B fluence responses were also studied (VvUVR8, VvHY5, VvCOP1 and VvCHS; PR genes and VvMAPK3; respectively). The genes associated with the low fluence response exhibited transcriptional regulation in line with reports from other species, while the PR genes and VvMAPK3 only appeared to be responsive in a high UV‐B fluence environment. Together, these data supports the view flavonol biosynthesis in grape is stimulated predominantly through the low fluence UV‐B response pathway.
This research investigates the effect of development and UV‐B on flavonols and the regulation of gene activity in Vitis vinifera L. var. Sauvignon blanc grapes. Results suggest flavonol biosynthesis and gene activity are stimulated by UV‐B and the low fluence UV‐B pathway is a major determinant of this response. There is also a strong influence of development on gene expression. This is the first research to analyse signal transduction associated with the recently discovered UV‐B photoreceptor UVR8 and makes a substantial contribution to understanding UV‐B responses in an important commercial species.
Methoxypyrazines are present in the grapes of certain Vitis vinifera varieties including Sauvignon blanc and contribute herbaceous/green aromas to wine. Environmental factors such as light exposure ...and temperature can influence methoxypyrazine levels, and viticultural interventions such as canopy manipulation have the ability to reduce methoxypyrazine accumulation in grapes. We assessed methoxypyrazine levels and showed that leaf removal significantly reduces accumulation in Sauvignon blanc grapes. The main effect of reducing methoxypyrazines was preveraison, as postveraison treatments had no effect on concentrations at harvest. Methoxypyrazine concentrations in controls peaked preveraison and decreased through harvest. Dilution due to an increase in berry weight was found to be the major driver of decreasing concentrations, as methoxypyrazine levels on a per berry basis were found to increase through development in two of three seasons. In the one year of our study that showed contrasting results, analyses of weather data indicate that warmer than average temperatures appear to be the principal factor affecting the berries’ ability to accumulate and retain methoxypyrazines. We also explored the expression of potential biosynthetic O-methyltransferase genes VvOMT1, VvOMT2, and VvOMT3; no significant differences were observed with respect to effect of leaf removal and light exposure.
•first comparison of controlled and vineyard environments to interpret flavonoid biosynthesis in response to high/low UV-B fluence.•controlled environments and vineyard trials present similar UV-B ...responses in flavonol accumulation and gene expression.•PR genes were not responsive to the natural UV-B fluence but regulated by berry development.
UV-B radiation is an environmental challenge affecting a number of metabolic functions in plants. Plants protect themselves from this potentially damaging radiation through synthesising UV-absorbing compounds such as flavonoids. This study aims to investigate the effect of UV-B on flavonoid biosynthesis in Sauvignon blanc grapes. In particular, a comparison has been made between controlled environment (CE) and vineyard trials to better understand molecular mechanisms of low/high fluence UV-B responses and how the results relate to each other in the context of flavonoid biosynthesis. Following exposure to supplemental UV-B in the CE, both flavonols and gene expression exhibited UV-B induced response. Flavonols, particularly quercetin/kaempferol 3-O-glycosides were increased at distinct stages of berry development. All genes measured showed a significant developmental regulation. VvFLS4, VvCHS1, VvMYB12, VvHY5 and PR (VvTL1 and VvChi4A/4B) increased due to UV-B in the CE experiments. However, PR were not responsive to the natural UV-B fluence in vineyard but were significantly induced at later stages of development. Overall, despite very different conditions in the CE and vineyard the majority of UV-B induced responses are similar. Only PR activities in the CE cabinets reflect a higher fluence stress response that is not reflected in the natural lower UV-B fluence environment.
Belclare and Cambridge are prolific sheep breeds, the origins of which involved selecting ewes with exceptionally high litter
size records from commercial flocks. The variation in ovulation rate in ...both breeds is consistent with segregation of a gene
(or genes) with a large effect on this trait. Sterile ewes, due to a failure of normal ovarian follicle development, occur
in both breeds. New naturally occurring mutations in genes for the oocyte-derived growth factors growth differentiation factor
9 (GDF9) and bone morphogenetic protein 15 (BMP15) are described. These mutations are associated with increased ovulation
rate in heterozygous carriers and sterility in homozygous carriers in both breeds. This is the first time that a mutation
in the gene for GDF9 has been found that causes increased ovulation rate and infertility in a manner similar to inactivating
mutations in BMP15, and shows that GDF9 is essential for normal folliculogenesis in sheep. Furthermore, it is shown, for the
first time in any species, that individuals with mutations in both GDF9 and BMP15 have a greater ovulation rate than sheep with either of the mutations separately.
The increase of temperature due to climate change at different phenological stages of grapevine has already been demonstrated to affect accumulation of primary and secondary metabolites in grape ...berries. This has a significant implication for Pinot noir especially in New Zealand context as these compounds can have direct and indirect effects on wine quality.
This study investigates how varying bunch microclimate through changes in temperature applied at veraison stage can affect: fresh weight, total soluble solids, the accumulation of anthocyanins, total phenolics and amino acids of the grape berries. This was studied over two growing seasons (2018/19 and 2019/20) with Pinot noir vines being grown at two different temperatures in controlled environment (CE) chambers. The vines were exposed to 800 µmol/m2/s irradiance with diurnal changes in day (22°C or 30°C) and night (15°C) temperatures. This experimental set up enabled us to determine the accumulation of these metabolite at harvest (both seasons) and throughout berry development (second season).
The results showed that berry weight was not influenced by temperature increase. The total soluble solids (TSS) were significantly increased at 30°C, however, this was not at the expense of berry weight (i.e., water loss). Anthocyanin content was reduced at higher temperature in the first season but there was no change in phenolic content in response to temperature treatments in either season. The concentrations of total amino acids at harvest increased in response to the higher temperature in the second season only. In addition, in the time course analysis of the second season, the accumulation of amino acids was increased at mid-ripening and ripening stage with the increased temperature. Significant qualitative changes in amino acid composition specifically the α-ketoglutarate family (i.e., glutamine, arginine, and proline) were found between the two temperatures.
This study is the first to provide detailed analysis and quantification of individual amino acids and phenolics in Pinot noir in response to changes in temperature applied at veraison which could aid to develop adaptation strategies for viticulture in the future.
Inflammation and matrix degradation are the hallmarks of high-risk atherosclerosis that leads to myocardial infarction and stroke. Toll-like receptors (TLRs), key players in innate immunity, are ...upregulated in atherosclerotic lesions, but their functional role in human atherosclerosis is unknown. We explored the effects of blocking TLR-2, TLR-4, and myeloid differentiation primary response gene 88 (MyD88), a signaling adaptor shared by most TLRs and interleukin-1 receptor (IL-1R), in an in vitro model of human atherosclerosis.
Carotid endarterectomies were obtained from patients with symptomatic carotid disease. Cells were isolated via enzymatic tissue dissociation and cultured in the presence or absence of TLR signaling blockers. A dominant-negative form of MyD88 (MyD88(DN)) decreased the production of monocyte chemotactic protein-1/CCL2 (P=0.000), IL-8/CXCL8 (P=0.006), IL-6 (P=0.002), matrix metalloproteinase-1 (MMP-1; P=0.002), and MMP-3 (P=0.000), as well as nuclear factor-kappaB activation (P<0.05) in atheroma cell cultures. IL-1R antagonist, TLR-4 blocking antibodies, or overexpression of a dominant-negative form of the TLR-4 signaling adaptor TRIF-related adaptor molecule reduced nuclear factor-kappaB activity but did not have a broad impact on the production of the mediators studied. In contrast, TLR-2 neutralizing antibodies inhibited nuclear factor-kappaB activation (P<0.05) and significantly reduced monocyte chemotactic protein-1/CCL2 (P=0.000), IL-8/CXCL8 (P=0.009), IL-6 (P=0.000), and MMP-1 (P=0.000), MMP-2 (P=0.004), MMP-3 (P=0.000), and MMP-9 (P=0.006) production.
Our data indicate that TLR-2 signaling through MyD88 plays a predominant role in inflammation and matrix degradation in human atherosclerosis. TLR-2 blockade may represent a therapeutic strategy for atherosclerosis and its complications.
Molecular characterisation of vulnerable atherosclerosis is necessary for targeting functional imaging and plaque-stabilising therapeutics. Inflammation has been linked to atherogenesis and the ...development of high-risk plaques. We set to quantify cytokine, chemokine and matrix metalloproteinase (MMP) protein production in cells derived from carotid plaques to map the inflammatory milieu responsible for instability. Carotid endarterectomies from carefully characterised symptomatic (n=35) and asymptomatic (n=32) patients were enzymatically dissociated producing mixed cell type atheroma cell suspensions which were cultured for 24 hours. Supernatants were interrogated for 45 analytes using the Luminex 100 platform. Twenty-nine of the 45 analytes were reproducibly detectable in the majority of donors. The in vitro production of a specific network of mediators was found to be significantly higher in symptomatic than asymptomatic plaques, including: tumour necrosis factor α, interleukin (IL) 1β, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), CCL5, CCL20, CXCL9, matrix metalloproteinase (MMP)-3 and MMP-9. Ingenuity pathway analysis of differentially expressed analytes between symptomatic and asymptomatic patients identified a number of key biological pathways (p< 10(-25)). In conclusion, the carotid artery plaque culprit of ischaemic neurological symptoms is characterised by an inflammatory milieu favouring inflammatory cell recruitment and pro-inflammatory macrophage polarisation.
Recent discoveries that high prolificacy in sheep carrying the Booroola gene ( FecB ) is the result of a mutation in the BMPIB receptor and high prolificacy in Inverdale sheep ( FecX I ) is the ...result of a mutation in the BMP15 oocyte-derived growth factor gene have allowed direct marker tests to be developed for FecB and FecX I . These tests were carried out in seven strains of sheep (Javanese, Thoka, Woodlands, Olkuska, Lacaune, Belclare, and Cambridge)
in which inheritance patterns have suggested the presence of major genes affecting prolificacy and in the prolific Garole
sheep of India, which have been proposed as the ancestor of Australian Booroola Merinos. The FecB mutation was found in the Garole and Javanese sheep but not in Thoka, Woodlands, Olkuska, Lacaune, Belclare, and Cambridge
sheep. None of the sheep tested had the FecX I mutation. These findings present strong evidence to support historical records that the Booroola gene was introduced into
Australian flocks from Garole (Bengal) sheep in the late 18th century. It is unknown whether Javanese Thin-tailed sheep acquired
the Booroola gene directly from Garole sheep from India or via Merinos from Australia. The DNA mutation test for FecB will enable breeding plans to be developed that allow the most effective use of this gene in Garole and Javanese Thin-tailed
sheep and their crosses.
Abstract
The aim of this research was to determine the effect of development and
UV
‐
B
on flavonols and the regulation of gene activity in
V
itis vinifera
L
. var.
S
auvignon blanc grapes. ...Particular emphasis was placed on gene activity associated with the low and high fluence
UV
‐
B
responses. Flavonols, particularly quercetin and kaempferol glycosides, increased substantially upon fruit exposure due to
UV
‐
B
, with spatial analysis locating the changes to the berry skin. Of five
VvFLS
genes in grapes, two (
VvFLS4
and
5
) were found to be transcriptionally active, with
VvFLS4
also being responsive to
UV
‐
B
but
VvFLS5
was not. Of the transcription factors known to regulate
FLS
(
VvMYB12
,
VvMYCA1
and
VvWDRs
), only
VvMYB12
was found to be responsive to
UV
‐
B
.
A number of candidate genes associated with the low and high
UV
‐
B
fluence responses were also studied (
VvUVR8
,
VvHY5
,
VvCOP1
and
VvCHS
;
PR
genes and
VvMAPK3
; respectively). The genes associated with the low fluence response exhibited transcriptional regulation in line with reports from other species, while the
PR
genes and
VvMAPK3
only appeared to be responsive in a high
UV
‐
B
fluence environment. Together, these data supports the view flavonol biosynthesis in grape is stimulated predominantly through the low fluence
UV
‐
B
response pathway.
This research investigates the effect of development and
UV‐B
on flavonols and the regulation of gene activity in
V
itis vinifera
L
. var. Sauvignon blanc grapes. Results suggest flavonol biosynthesis and gene activity are stimulated by
UV‐B
and the low fluence
UV‐B
pathway is a major determinant of this response. There is also a strong influence of development on gene expression. This is the first research to analyse signal transduction associated with the recently discovered
UV‐B
photoreceptor
UVR8
and makes a substantial contribution to understanding
UV‐B
responses in an important commercial species.
Belclare and Cambridge are prolific sheep breeds, the origins of which involved selecting ewes with exceptionally high litter size records from commercial flocks. The variation in ovulation rate in ...both breeds is consistent with segregation of a gene (or genes) with a large effect on this trait. Sterile ewes, due to a failure of normal ovarian follicle development, occur in both breeds. New naturally occurring mutations in genes for the oocyte-derived growth factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are described. These mutations are associated with increased ovulation rate in heterozygous carriers and sterility in homozygous carriers in both breeds. This is the first time that a mutation in the gene for GDF9 has been found that causes increased ovulation rate and infertility in a manner similar to inactivating mutations in BMP15, and shows that GDF9 is essential for normal folliculogenesis in sheep. Furthermore, it is shown, for the first time in any species, that individuals with mutations in both GDF9 and BMP15 have a greater ovulation rate than sheep with either of the mutations separately.
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