Physiological, biochemical and transcriptional analysis of onion bulbs during storage

Abstract: During the transition from endo-dormancy to eco-dormancy and subsequent growth, the onion bulb undergoes the transition from sink organ to source, to sustain cell division in the meristematic tissue. The mechanisms controlling these processes are not fully understood. Here, a detailed analysis of whole onion bulb physiological, biochemical and transcriptional changes in response to sprouting is reported, enabling a better knowledge of the mechanisms regulating post-harvest onion sprout development. Biochemical and physiological analyses were conducted on different cultivars ('Wellington', 'Sherpa' and 'Red Baron') grown at different sites over 3 years, cured at different temperatures (20, 24 and 28 degrees C) and stored under different regimes (1, 3, 6 and 6 1 degrees C). In addition, the first onion oligonucleotide microarray was developed to determine differential gene expression in onion during curing and storage, so that transcriptional changes could support biochemical and physiological analyses. There were greater transcriptional differences between samples at harvest and before sprouting than between the samples taken before and after sprouting, with some significant changes occurring during the relatively short curing period. These changes are likely to represent the transition from endo-dormancy to sprout suppression, and suggest that endo-dormancy is a relatively short period ending just after curing. Principal component analysis of biochemical and physiological data identified the ratio of monosaccharides (fructose and glucose) to disaccharide (sucrose), along with the concentration of zeatin riboside, as important factors in discriminating between sprouting and pre-sprouting bulbs. These detailed analyses provide novel insights into key regulatory triggers for sprout dormancy release in onion bulbs and provide the potential for the development of biochemical or transcriptional markers for sprout initiation. Evidence presented herein also suggests there is no detrimental effect on bulb storage life and quality caused by curing at 20 degrees C, producing a considerable saving in energy and costs.

Shoot yield drives phosphorus use efficiency in Brassica oleracea and correlates with root architecture traits

The environmental and financial costs of using inorganic phosphate fertilizers to maintain crop yield and quality are high. Breeding crops that acquire and use phosphorus (P) more efficiently could reduce these costs. The variation in shoot P concentration (shoot-P) and various measures of P use efficiency (PUE) were quantified among 355 Brassica oleracea L. accessions, 74 current commercial cultivars, and 90 doubled haploid (DH) mapping lines from a reference genetic mapping population. Accessions were grown at two or more external P concentrations in glasshouse experiments; commercial and DH accessions were also grown in replicated field experiments. Within the substantial species-wide diversity observed for shoot-P and various measures of PUE in B. oleracea, current commercial cultivars have greater PUE than would be expected by chance. This may be a consequence of breeding for increased yield, which is a significant component of most measures of PUE, or early establishment. Root development and architecture correlate with PUE; in particular, lateral root number, length, and growth rate. Significant quantitative trait loci associated with shoot-P and PUE occur on chromosomes C3 and C7. These data provide information to initiate breeding programmes to improve PUE in B. oleracea.

Relationships between yield and mineral concentrations in potato tubers

There is concern that modern cultivars and/or agronomic practices have resulted in reduced concentrations of mineral elements essential to human nutrition in edible crops. Increased yields are often associated with reduced concentrations of mineral elements in produce, and a number of recent studies have indicated that, when grown under identical conditions, the concentrations of several mineral elements are lower in genotypes yielding more grain or shoot biomass than in older, lower-yielding genotypes. Potato is a significant crop, grown worldwide, yet few studies have investigated whether increasing yields, through agronomy or breeding, affects the concentrations of mineral elements in tubers. This article examines the hypothesis that increasing yields, either by the application of mineral fertilizers and/or by growing higher-yielding varieties, leads to decreased concentrations of mineral elements in tubers. It reports that the application of fertilizers influences tuber elemental composition in a complex manner, presumably as a consequence of soil chemistry and interactions between mineral elements within the plant, that considerable variation exists between potato genotypes in the concentrations of mineral elements in their tubers, and that, like in other crops, higher-yielding genotypes occasionally have lower concentrations of some mineral elements in their edible tissues than lower-yielding genotypes.

Spatial patterns of gluten protein and polymer distribution in wheat grain

The starchy endosperm is the major storage tissue in the mature wheat grain and exhibits quantitative and qualitative gradients in composition, with the outermost cell layers being rich in protein, mainly gliadins, and the inner cells being low in protein but enriched in high-molecular-weight (HMW) subunits of glutenin. We have used sequential pearling to produce flour fractions enriched in particular cell layers to determine the protein gradients in four different cultivars grown at two nitrogen levels. The results show that the steepness of the protein gradient is determined by both genetic and nutritional factors, with three high-protein breadmaking cultivars being more responsive to the N treatment than a low-protein cultivar suitable for livestock feed. Nitrogen also affected the relative abundances of the three main classes of wheat prolamins: the sulfur-poor ω-gliadins showed the greatest response to nitrogen and increased evenly across the grain; the HMW subunits also increased in response to nitrogen but proportionally more in the outer layers of the starchy endosperm than near the core, while the sulfur-rich prolamins showed the opposite trend.

Expression of epitope-tagged LMW glutenin subunits in the starchy endosperm of transgenic wheat and their incorporation into glutenin polymers

The low-molecular-weight (LMW) glutenin subunits are components of the highly cross-linked glutenin polymers that confer viscoelastic properties to gluten and dough. They have both quantitative and qualitative effects on dough quality that may relate to differences in their ability to form the inter-chain disulphide bonds that stabilise the polymers. In order to determine the relationship between dough quality and the amounts and properties of the LMW subunits, we have transformed the pasta wheat cultivars Svevo and Ofanto with three genes encoding proteins, which differ in their numbers or positions of cysteine residues. The transgenes were delivered under control of the high-molecular-weight (HMW) subunit 1Dx5 gene promoter and terminator regions, and the encoded proteins were C-terminally tagged by the introduction of the c-myc epitope. Stable transformants were obtained with both cultivars, and the use of a specific antibody to the c-myc epitope tag allowed the transgene products to be readily detected in the complex mixture of LMW subunits. A range of transgene expression levels was observed. The addition of the epitope tag did not compromise the correct folding of the trangenic subunits and their incorporation into the glutenin polymers. Our results demonstrate that the ability to specifically epitope-tag LMW glutenin transgenes can greatly assist in the elucidation of their individual contributions to the functionality of the complex gluten system.

Morpho-physiological characterization of spring wheat genotypes under drought stress

Water-deficit is a severe abiotic stress and major constraint to wheat productivity with effect on plant growth and development. The objective of this study was to characterize drought tolerant and susceptible spring wheat cultivars on the basis of physiological and yield attributes. The experiment was comprised of two irrigation regimes i.e. irrigated and 65% drought stress and ten wheat cultivars viz. Anmol, Moomal, Sarsabz, Bhittai, Pavon, SKD-1, TD-1, Kiran, Marvi and Mehran. Results indicated significant effect of water stress on stomatal dimension, stomatal conductance, relative leaf water content and grain yield with no effect on stomatal density. The irrigation × cultivars interaction was non-significant for grain yield only. Cultivars like Anmol, Moomal, Bhittai, Sarsabz proved to be drought tolerant with smaller stomatal dimensions, less stomatal conductance and more relative water content under water stress and produced higher grain yield. While decrease in relative water contents and grain yield, and increase in stomatal attributes was observed in drought susceptible cultivars such as Marvi, TD-1 and SKD-1 hence proved to be drought susceptible.

Temporal sensitivity of rice seed development from spikelet fertility to viable mature seed to extreme-temperature

Extreme temperature during reproductive development affects rice (Oryza sativa L.) yield and seed quality. A controlled-environment reciprocal-transfer experiment was designed where plants from two japonica cultivars were grown at 28/24 ⁰C and moved to 18/14 ⁰C and vice versa, or from 28/24 to 38/34 ⁰C and vice versa, for 7-d periods to determine the respective temporal pattern of sensitivity of spikelet fertility, yield, and seed viability to each temperature extreme. Spikelet fertility and seed yield per panicle were severely reduced by extreme temperature in the 14 d period prior to anthesis; and both cultivars were affected at 38/34 ⁰C while only cv. Gleva was affected at 18/14 ºC. The damage was greater the earlier the panicles were stressed within this period. Later-exserted panicles compensated only partly for yield loss. Seed viability was significantly reduced by 7-d exposure to 38/34 ⁰C or 18/14 ⁰C at 1 to 7 and 1 to 14 d after anthesis, respectively, in cv. Gleva. Cultivar Taipei 309 was not affected by 7 d exposure at 18/14 ⁰C; and no consistent temporal pattern of sensitivity was evident at 38/34 ⁰C. Hence, brief exposure to low or high temperature was most damaging to spikelet fertility and yield 14 to 7 d before anthesis, coinciding with microsporogenesis; and it was almost as damaging around anthesis. Seed viability was most vulnerable to low or high temperature in the 7 or 14 d after anthesis, when histodifferentiation occurs.

High night temperature induces contrasting responses for spikelet fertility, spikelet tissue temperature, flowering characteristics and grain quality in rice

Climate change is increasing night temperature (NT) more than day temperature (DT) in rice-growing areas. Effects of combinations of NT (24-35°C) from microsporogenesis to anthesis at one or more DT (30 or 35°C) at anthesis on rice spikelet fertility, temperature within spikelets, flowering pattern, grain weight per panicle, amylose content and gel consistency were investigated in contrasting rice cultivars under controlled environments. Cultivars differed in spikelet fertility response to high NT, with higher fertility associated with cooler spikelets (P < 0.01). Flowering dynamics were altered by high NT and a novel high temperature tolerance complementary mechanism, shorter flower open duration in cv. N22, was identified. High NT reduced spikelet fertility, grain weight per panicle, amylose content and gel consistency, whereas high DT reduced only gel consistency. Night temperature >27°C was estimated to reduce grain weight. Generally, high NT was more damaging to grain weight and selected grain quality traits than high DT, with little or no interaction between them. The critical tolerance and escape traits identified, i.e. spikelet cooling, relatively high spikelet fertility, earlier start and peak time of anthesis and shorter spikelet anthesis duration can aid plant breeding programs targeting resilience in warmer climates.

Antibiosis in wheat interacts with crowding stress to affect Metopolophiumdirhodum development and susceptibility to malathion

We used a laboratory study to compare the performance of rose-grain aphid, Metopolophium dirhodum(Walker)(Hemiptera:Aphididae),onthewheatcultivars‘Huntsman’(susceptible)and‘Rapier’ (partiallyresistant)inbothlowdensity(uncrowded)andhighdensity(crowded)coloniesandexamined the consequences for aphid susceptibility to malathion. Adult apterae that developed on Rapier wheat had their mean relative growth rate (MRGR) reduced by 6 and 9% under uncrowded and crowded conditions, respectively, whereas the crowding treatment reduced MRGR by 3%, but only in Rapier aphids. Rapier resistance also reduced adult dry weight by 13 and 14% under crowded and uncrowded conditions, respectively, whereas crowding reduced it by 34 and 35% in Rapier and Huntsman aphids, respectively. Development on Rapier substantially reduced the topical LC50 of malathion by 37.8 and 34.8% under crowded and uncrowded conditions, suggesting that plant antibiosis increased malathion susceptibility. By comparison, crowding only reduced the LC50 by 29.5 and 26.0% on Huntsman and Rapier. The LD50 data showed that reductions on aphid body size on Rapier and through crowding did not fully explain the differences in LC50. This was particularly in the values for crowded aphids that were actually 80% higher than for uncrowded ones. Thi sapparent tolerance of crowded aphids, however, may partly be due to loss of insecticide from small aphids at dosing. Evidence of synergy between plant resistance and insecticide susceptibility raisest he possibility of using reduced concentrations of pesticides to control aphids on resistant crop cultivars, with diminished impacts on non-target and beneficial species important in integrated pest management(IPM)program

Behavioural responses of the seven-spot ladybird Coccinella septempunctata to plant headspace chemicals collected from four crop Brassicas and Arabidopsis thaliana, infested with Myzus persicae

1 Insects using olfactory stimuli to forage for prey/hosts are proposed to encounter a ‘reliability–detectability problem’, where the usability of a stimulus depends on its reliability as an indicator of herbivore presence and its detectability. 2 We investigated this theory using the responses of female seven-spot ladybirds Coccinella septempunctata (Coleoptera: Coccinellidae) to plant headspace chemicals collected from the peach-potato aphid Myzus persicae and four commercially available Brassica cultivars; Brassica rapa L. cultivar ‘turnip purple top’, Brassica juncea L. cultivar ‘red giant mustard’, Brassica napus L. cultivar ‘Apex’, Brassica napus L. cultivar ‘Courage’ and Arabidopsis thaliana. For each cultivar/species, responses to plants that were undamaged, previously infested by M. persicae and infested with M. persicae, were investigated using dual-choice Petri dish bioassays and circular arenas. 3 There was no evidence that ladybirds responded to headspace chemicals from aphids alone. Ladybirds significantly preferred headspace chemicals from B. napus cv. Apex that were undamaged compared with those from plants infested with aphids. For the other four species/cultivars, there was a consistent trend of the predators being recorded more often in the half of the Petri dish containing plant headspace chemicals from previously damaged and infested plants compared with those from undamaged ones. Furthermore, the mean distance ladybirds walked to reach aphid-infested A. thaliana was significantly shorter than to reach undamaged plants. These results suggest that aphid-induced plant chemicals could act as an arrestment or possibly an attractant stimulus to C. septempunctata. However, it is also possible that C. septempunctata could have been responding to aphid products, such as honeydew, transferred to the previously damaged and infested plants. 4 The results provide evidence to support the ‘reliability–detectability’ theory and suggest that the effectiveness of C. septempunctata as a natural enemy of aphids may be strongly affected by which species and cultivar of Brassica are being grown.

Foraging strategies of natural enemies of the peach-potato aphid Myzus persicae

The foraging strategies of two natural enemies of the peach-potato aphid Myzus persicae: the seven-spot ladybird Coccinella septempunctata and the parasitoid wasp Diaeretiella rapae, were investigated. Specifically the roles of plant semiochemicals in the location of plants infested with M. persicae by these natural enemies were examined. I investigated the olfactory responses of female C. septempunctata to volatiles collected from M. persicae and four Brassica cultivars; Brassica rapa, B. juncea, B. napus cultivar ‘Apex’ and B. napus cultivar ‘Courage’ and wild-type Arabidopsis thaliana that were: undamaged, previously infested by M. persicae and infested with M. persicae. C. septempunctata showed no attraction to volatiles from M. persicae alone. C. septempunctata significantly changed its searching behaviour in response to plant volatiles from B. rapa, B. napus cv. ‘Apex’ and Arabidopsis infested with M. persicae. C. septempunctata was also found to display a significant turning bias when foraging on a branching horizontal wire stem. A model was developed to investigate how turning biases affect the foraging efficiency of C. septempunctata in dichotomous branched environments. Simulations using this model indicated that turning biases could potentially increase searching efficiency. D. rapae showed a significant preference for volatiles from M. persicae infested wild-type Arabidopsis but no preference to volatiles from M. persicae alone or M. persicae honeydew. Volatile emissions by Arabidopsis were shown to be localised to the area of aphid-infestation rather than systemic. Using gas chromatography plants infested with M. persicae were shown to emit a quantitatively different volatile blend than undamaged plants. In experiments with jasmonate mutants of Arabidopsis the jasmonate (octadecanoid) wound response pathway was implicated as being important for the production of M. persicae induced volatiles, attractive to D. rapae. Other wound response pathways were also found to be involved in the production of the full blend of M. persicae induced volatiles.

Influence of temperature during grain filling on gluten viscoelastic properties and gluten protein composition

BACKGROUND The aim of this study was to investigate the effects of low to moderate temperatures on gluten functionality and gluten protein composition. Four spring wheat cultivars were grown in climate chambers with three temperature regimes (day/night temperatures of 13/10, 18/15 and 23/20 °C) during grain filling. RESULTS The temperature strongly influenced grain weight and protein content. Gluten quality measured by maximum resistance to extension (Rmax) was highest in three cultivars grown at 13 °C. Rmax was positively correlated with the proportion of sodium dodecyl sulfate-unextractable polymeric proteins (%UPP). The proportions of ω-gliadins and D-type low-molecular-weight glutenin subunits (LMW-GS) increased and the proportions of α- and γ-gliadins and B-type LMW-GS decreased with higher temperature, while the proportion of high-molecular-weight glutenin subunits (HMW-GS) was constant between temperatures. The cultivar Berserk had strong and constant Rmax between the different temperatures. CONCLUSION Constant low temperature, even as low as 13 °C, had no negative effects on gluten quality. The observed variation in Rmax related to temperature could be explained more by %UPP than by changes in the proportions of HMW-GS or other gluten proteins. The four cultivars responded differently to temperature, as gluten from Berserk was stronger and more stable over a wide range of temperature

Influence of temperature on the composition and polymerization of gluten proteins during grain filling in spring wheat (Triticum aestivum L.)

One Norwegian and one UK spring wheat cultivar, Bjarne and Cadenza, respectively, were grown in climate chambers to investigate the effects of lower to moderate temperatures during grain filling on the gluten quality. Two experiments were carried out with weekly fertilization until anthesis, while post-anthesis fertilization was applied in a third experiment. The proportions of different gluten proteins were affected by temperature in a similar manner for both cultivars when grown without post-anthesis fertilization. However, whereas low temperature strongly decreased %UPP for Cadenza, Bjarne had high %UPP at all temperature regimes. The results indicated that the assembly of glutenin polymers in Bjarne was less sensitive to variation in temperature than in Cadenza. Thus, our results suggested that the temperature influenced the proportion of different gluten proteins in both cultivars, while its effects on the assembly of the glutenin polymers were cultivar dependent. The duration of grain filling was longer at the lower temperatures, and this was associated with increased grain weight. Temperature had little effect on the amount of protein accumulated per grain, thus the proportion of proteins was strongly decreased at lower temperatures. This was to some extent, but not fully counteracted by post-anthesis fertilization.

Tales of the unexpected: research discoveries that occurred by accident

Data from four experimental research projects are presented which have in common that unexpected results caused a change in direction of the research. A plant growth accelerator caused the appearance of white black bean aphids, a synthetic pyrethroid suspected of enhancing aphid reproduction proved to enhance plant growth, a chance conversation with a colleague initiated a search for fungal DNA in aphids, and the accidental invasion of aphid cultures by a parasitoid reversed the aphid population ranking of two Brussels sprout cultivars. This last result led to a whole series of studies on the plant odour preferences of emerging parasitoids which in turn revealed the unexpected phenomenon that chemical cues to the maternal host plant are left with the eggs at oviposition. It is pointed out that, too often, researchers fail to follow up unexpected results because they resist accepting flaws in their hypotheses; also that current application criteria for research funding make it hard to accommodate unexpected findings.

Susceptibility of faba bean (Vicia faba L.) to heat stress during floral development and anthesis

Experiments were conducted over two years to quantify the response of faba bean (Vicia faba L.) to heat stress. Potted winter faba bean plants (cv. Wizard) were exposed to temperature treatments (18/10; 22/14; 26/18; 30/22; 34/26°C day/night) for five days during floral development and anthesis. Developmental stages of all flowers were scored prior to stress, plants were grown in exclusion from insect pollinators to prevent pollen movement between flowers, and yield was harvested at an individual pod scale, enabling effects of heat stress to be investigated at a high resolution. Susceptibility to stress differed between floral stages, flowers were most affected during initial green-bud stages. Yield and pollen germination of flowers present before stress showed threshold relationships to stress, with lethal temperatures (t50) ~28°C and ~32°C, while whole plant yield showed a linear negative relationship to stress with high plasticity in yield allocation, such that yield lost at lower nodes was partially compensated at higher nodal positions. Faba bean has many beneficial attributes for sustainable modern cropping systems but these results suggest that yield will be limited by projected climate change, necessitating the development of heat tolerant cultivars, or improved resilience by other mechanisms such as earlier flowering times.