Everard Edwards

Seed Persistence: A Correlation Between Seed Longevity in the Soil and Ortho-Dihydroxyphenol Concentration

1. Ortho-dihydroxyphenol and cyanoglycoside concentrations were determined in seeds of over 80 species, drawn largely from the native British flora but including a number of weeds and other species of agricultural significance. 2. All seeds examined contained at least trace amounts of hydroxyphenols and threequarters released hydrogen cyanide on hydrolysis. A highly significant correlation was found between (a) seed persistence in the soil and ortho-dihydroxyphenol concentration and (b) between protein content, as a measure of the nutritional value of the seed, and ortho-dihydroxyphenol content. No such relation was found with cyanoglycosides. 3. Particularly high concentrations of ortho-dihydroxyphenols were found throughout species within the order Myrtales (Onagraceae and relatives), all with little or no seed endosperm. 4. From this broad survey we conclude that ortho-dihydroxyphenols, as a class, play an important role in maintaining seed persistence in the soil, probably by deferring or decreasing the rate of decomposition by microbes as well as by defending against herbivory, and that this is one important mechanism, along with morphological adaptations, which confers longevity and persistence on seeds buried in cool temperate and relatively moist soil conditions.

Nitrogen in cell walls of sclerophyllous leaves accounts for little of the variation in photosynthetic nitrogen-use efficiency

Photosynthetic rate per unit nitrogen generally declines as leaf mass per unit area (LMA) increases. To determine how much of this decline was associated with allocating a greater proportion of leaf nitrogen into cell wall material, we compared two groups of plants. The first group consisted of two species from each of eight genera, all of which were perennial evergreens growing in the Australian National Botanic Gardens (ANBG). The second group consisted of seven Eucalyptus species growing in a greenhouse. The percentage of leaf biomass in cell walls was independent of variation in LMA within any genus, but varied from 25 to 65% between genera. The nitrogen concentration of cell wall material was 0.4 times leaf nitrogen concentration for all species apart from Eucalyptus, which was 0.6 times leaf nitrogen concentration. Between 10 and 30% of leaf nitrogen was recovered in the cell wall fraction, but this was independent of LMA. No trade-off was observed between nitrogen associated with cell walls and the nitrogen allocated to ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco). Variation in photosynthetic rate per unit nitrogen could not be explained by variation in cell wall nitrogen.

Expression of ABA synthesis and metabolism genes under different irrigation strategies and atmospheric VPDs is associated with stomatal conductance in grapevine (Vitis vinifera L. cv Cabernet Sauvignon)

The influence of different levels of irrigation and of variation in atmospheric vapour pressure deficit (VPD) on the synthesis, metabolism, and transport of abscisic acid (ABA) and the effects on stomatal conductance were examined in field-grown Cabernet Sauvignon grapevines. Xylem sap, leaf tissue, and root tissue were collected at regular intervals during two seasons in conjunction with measurements of leaf water potential (Ψleaf) and stomatal conductance (gs). The different irrigation levels significantly altered the Ψleaf and gs of the vines across both seasons. ABA abundance in the xylem sap was correlated with gs. The expression of genes associated with ABA synthesis, NCED1 and NCED2, was higher in the roots than in the leaves throughout and highest in the roots in mid January, a time when soil moisture declined and VPD was at its highest. Their expression in roots was also inversely related to the levels of irrigation and correlated with ABA abundance in the roots, xylem sap, and leaves. Three genes encoding ABA 8’-hydroxylases were isolated and their identities confirmed by expression in yeast cells. The expression of one of these, Hyd1, was elevated in leaves when VPD was below 2.0–2.5 kPa and minimal at higher VPD levels. The results provide evidence that ABA plays an important role in linking stomatal response to soil moisture status and that changes in ABA catabolism at or near its site of action allows optimization of gas exchange to current environmental conditions.

Is there a link between greening and light-enhanced glycoalkaloid accumulation in potato (Solanum tuberosumL) tubers?

Potato (Solanum tuberosum L) tubers (cvs Pentland Dell and Record) were treated with the chlorophyll biosynthesis inhibitors 4-amino-5-fluoropentanoic acid (AFPA) and 3-amino-2,3-dihydrobenzoic acid (gabaculine), and subsequently exposed to daylight for up to 10 days prior to pigment and glyco-alkaloid analysis. AFPA inhibited the accumulation of total chlorophyll (Chl) by between 50 and 70% in both cultivars throughout the duration of light exposure. The synthesis of Chl b was inhibited by over 80% in both cultivars. Neither inhibitor had a significant effect on light-enhanced glycoalkaloid accumulation. It is concluded that there is no direct metabolic link between Chl and glycoalkaloid biosynthesis.

Anthocyanin biosynthesis is differentially regulated by light in the skin and flesh of white-fleshed and teinturier grape berries

AbstractMain conclusionLight exclusion reduces the concentration and modifies the composition of grape anthocyanins, by altering the expression of genes involved in anthocyanin biosynthesis and transport, in a cultivar- and tissue-specific manner. Unlike most grapes, teinturier grapes accumulate anthocyanins both in skin and flesh. However, the concentration and composition of anthocyanins in both tissues differ, providing a valuable system to study tissue-specific regulation of anthocyanin synthesis. Furthermore, little is known about the mechanisms controlling the sensitivity of anthocyanin accumulation to light. Here, light was excluded from Gamay (white-fleshed) and Gamay Fréaux (teinturier mutant) berries throughout berry development. Under light-exposed conditions, the skin of Gamay Fréaux accumulated the highest level of anthocyanins, followed by the skin of Gamay, while the pulp of Gamay Fréaux had much lower anthocyanins than the skins. Network analysis revealed the same order on the number of significant correlations among metabolites and transcripts in the three colored tissues, indicating a higher connectivity that reflects a higher efficiency of the anthocyanin pathway. Compared to light conditions, light exclusion reduced the total amount of anthocyanins, most severely in the skin of Gamay and to a lesser extent in the flesh and skin of Gamay Fréaux. Coordinated decrease in the transcript abundance of structural, regulatory and transporter genes by light exclusion correlated with the reduced anthocyanin concentration in a cultivar- and tissue-specific manner. Moreover, light exclusion increased the ratio of dihydroxylated to trihydroxylated anthocyanins, in parallel with F3′H and F3′5′H transcript amounts. Sugars and ABA only play a limited role in the control of anthocyanin synthesis in the berries, in contrast with what has been described in cell suspensions. This study provides novel insights into the regulation of anthocyanin in wild type and teinturier cultivars.

The effect of prior storage on the potential of potato tubers (Solanum tuberosum L) to accumulate glycoalkaloids and chlorophylls during light exposure, including artificial neural network modelling

Potato tubers of four varieties (Brodick, King Edward, Pentland Dell and Record) were stored under commercial conditions and exposed to light for up to 10 days after 0, 10, 20 and 30 weeks. These were analysed for photosynthetic pigment and glycoalkaloid content. There was no significant alteration in either tuber chlorophyll or glycoalkaloid content during dark storage. All four varieties greened in response to light exposure, but only three exhibited a significant increase in glycoalkaloid concentrations during this exposure. Storage duration did not significantly affect pigment accumulation. However, there was a marked effect of storage on the extent of glycoalkaloid accumulation. Tubers of all four varieties stored for more than 10 weeks did not accumulate glycoalkaloids to the same extent as fresh tubers. Indeed, Brodick and Record did not accumulate any glycoalkaloids in response to light after 30 weeks of storage. A number of artificial neural network models of the results were produced. These accurately modelled cultivars individually, but a model encompassing all the data was not successful at predicting cultivar differences.

Changes in Nutritional Value of Cyanogenic Trifolium repens Grown at Elevated Atmospheric CO2

Global food security in a changing climate depends on both the nutritive value of staple crops as well as their yields. Here, we examined the direct effect of atmospheric carbon dioxide on the toxicity of the important pasture crop, Trifolium repens L. (clover). Shoots of T. repens contain cyanogenic glycosides that break down to release toxic hydrogen cyanide when damaged. The ability of animals to tolerate cyanogenic compounds is dependent, in part, on their overall protein intake. We grew T. repens communities at ambient and approximately twice-ambient CO2 in a controlled environment greenhouse experiment. We found that the ratio of total cyanogenic glycosides to total protein ratio was nearly two times higher in leaves of T. repens grown at elevated CO2. This study highlights the importance of assessing the nutritive value of this and other plants in response to rising CO2 so that steps can be taken to address any adverse consequences for herbivores.

VitiCanopy: A Free Computer App to Estimate Canopy Vigor and Porosity for Grapevine

Leaf area index (LAI) and plant area index (PAI) are common and important biophysical parameters used to estimate agronomical variables such as canopy growth, light interception and water requirements of plants and trees. LAI can be either measured directly using destructive methods or indirectly using dedicated and expensive instrumentation, both of which require a high level of know-how to operate equipment, handle data and interpret results. Recently, a novel smartphone and tablet PC application, VitiCanopy, has been developed by a group of researchers from the University of Adelaide and the University of Melbourne, to estimate grapevine canopy size (LAI and PAI), canopy porosity, canopy cover and clumping index. VitiCanopy uses the front in-built camera and GPS capabilities of smartphones and tablet PCs to automatically implement image analysis algorithms on upward-looking digital images of canopies and calculates relevant canopy architecture parameters. Results from the use of VitiCanopy on grapevines correlated well with traditional methods to measure/estimate LAI and PAI. Like other indirect methods, VitiCanopy does not distinguish between leaf and non-leaf material but it was demonstrated that the non-leaf material could be extracted from the results, if needed, to increase accuracy. VitiCanopy is an accurate, user-friendly and free alternative to current techniques used by scientists and viticultural practitioners to assess the dynamics of LAI, PAI and canopy architecture in vineyards, and has the potential to be adapted for use on other plants.

ABA-mediated responses to water deficit separate grapevine genotypes by their genetic background

BackgroundABA-mediated processes are involved in plant responses to water deficit, especially the control of stomatal opening. However in grapevine it is not known if these processes participate in the phenotypic variation in drought adaptation existing between genotypes. To elucidate this question, the response to short-term water-deficit was analysed in roots and shoots of nine Vitis genotypes differing in their drought adaptation in the field. The transcript abundance of 12 genes involved in ABA biosynthesis, catabolism, and signalling were monitored, together with physiological and metabolic parameters related to ABA and its role in controlling plant transpiration.ResultsAlthough transpiration and ABA responses were well-conserved among the genotypes, multifactorial analyses separated Vitis vinifera varieties and V. berlandieri x V. rupestris hybrids (all considered drought tolerant) from the other genotypes studied. Generally, V. vinifera varieties, followed by V. berlandieri x V. rupestris hybrids, displayed more pronounced responses to water-deficit in comparison to the other genotypes. However, changes in transcript abundance in roots were more pronounced for Vitis hybrids than V. vinifera genotypes. Changes in the expression of the cornerstone ABA biosynthetic gene VviNCED1, and the ABA transcriptional regulator VviABF1, were associated with the response of V. vinifera genotypes, while changes in VviNCED2 abundance were associated with the response of other Vitis genotypes. In contrast, the ABA RCAR receptors were not identified as key components of the genotypic variability of water-deficit responses. Interestingly, the expression of VviSnRK2.6 (an AtOST1 ortholog) was constitutively lower in roots and leaves of V. vinifera genotypes and higher in roots of V. berlandieri x V. rupestris hybrids.ConclusionsThis study highlights that Vitis genotypes exhibiting different levels of drought adaptation differ in key steps involved in ABA metabolism and signalling; both under well-watered conditions and in response to water-deficit. In addition, it supports that adaptation may be related to various mechanisms related or not to ABA responses.

A Simple Microplate Assay to Quantify Nonstructural Carbohydrates of Grapevine Tissues

Knowledge of total nonstructural carbohydrates in plant material is useful for understanding the effects of environmental conditions and vineyard management decisions on grapevine physiology. While many techniques are described in the literature, varying in degree of sophistication, few have been validated in grapevines. We describe the miniaturization of a simple extraction method and microplate based colorimetric assay for the measurement of nonstructural carbohydrates in a range of grapevine tissues. The method provides quantitative results for soluble carbohydrates and starch in samples containing as low as 20 μg carbohydrate.