Conor O'Reilly

An in vivo root hair assay for determining rates of apoptotic-like programmed cell death in plants

In Arabidopsis thaliana we demonstrate that dying root hairs provide an easy and rapid in vivo model for the morphological identification of apoptotic-like programmed cell death (AL-PCD) in plants. The model described here is transferable between species, can be used to investigate rates of AL-PCD in response to various treatments and to identify modulation of AL-PCD rates in mutant/transgenic plant lines facilitating rapid screening of mutant populations in order to identify genes involved in AL-PCD regulation.

Proposed dates for lifting Sitka spruce planting stock for fresh planting or cold storage, based on physiological indicators

The relationship between physiological status and theability to withstand the stresses of lifting, coldstoring and planting was evaluated from 1992–1995 inSitka spruce (Picea sitchensis (Bong.) Carr.)2 + 1 transplants of Washington/northern Oregon originsgrowing in Ireland. Needle primordium initiationcontinued until later and cold hardiness developedmore slowly in the mild lifting season of 1994/95. The seasonal decline in shoot mitotic activity wasgenerally similar each year, with apices becominginactive in December. Cell divisions in roots ceasedin November or December. Shoots began to deharden inearly February each year, but the rates of dehardeningwere slow until March. Cell divisions in root apicesresumed in February each year, compared with March forshoot apices. The vitality of the seedlings followingcold storage was high for most lift dates except theOctober and April dates. The period of highest stressresistance (as indicated by cold hardiness levels) forlifting for immediate planting was from about lateNovember/early December to early March; shoots werecold hardy to about −20 °C (LT50) at thistime. Seedlings could be lifted and cold stored (1–2 °C)until May/June when shoots were cold hardyto about −30 °C or lower, which coincided withthe period from mid- to late December until March.

Stress resistance levels change little during dormancy in ash, sessile oak and sycamore seedlings

The physiological status of ash (Fraxinus excelsior L.), sessile oak (Quercus petraea (Matt.)) and sycamore (Acer pseudoplatanus L.) seedlings sampled from nurseries in Ireland between October and May, 1995/1996 and 1996/1997 was investigated. The results of heat treatments applied to taproots in vitro and evaluated using root electrolyte leakage (REL) indicated that sycamore was more easily damaged than the other species but lift date effects were small in all species. Dormancy release began later each year in ash (mid-November) than in oak or sycamore (late October to early November). The results of the REL and other data indicated that there was little change in stress resistance from November to about March (and April for some parameters). Sycamore was most active (highest shoot xylem pressure potential, REL, and root growth potential) during the winter, suggesting that adverse handling during this period is likely to reduce plant quality than in ash or oak. Field performance was good regardless of lifting/planting date.

Drying and soaking pretreatments affect germination in pedunculate oak

Abstract• Pedunculate oak (Quercus robur L.) acorns are recalcitrant so they are difficult to store. Therefore, deterioration in quality during storage may reduce seedling yield and quality. In an attempt to address this, the effects of drying and soaking treatments on acorn germination were assessed.• After harvesting on two occasions in October, the moisture content (MC) of acorns were adjusted using the following treatments: (1) fresh state or control (46–48% MC); (2) soaked in fresh state (46–48%); (3) dried (40–42%) (standard method); and (4) dried and then soaked (46–48%). The treated acorns were allowed to germinate at 15 °C, both before and after storage at −3 °C.• Treatment effects were generally consistent for each harvest date. Compared to the untreated controls, soaking alone and drying followed by soaking significantly increased germination both before and after storage. Storage reduced germination, except for the dried and soaked acorns.• The results confirmed that drying and soaking improved acorn germination, but this was not mediated through an effect on acorn MC. In a separate nursery experiment, acorns given this treatment resulted in higher seedling yields and better quality plants than those given the standard treatment (dried to 40–42% MC).Résumé• Les glands du chêne pédonculé (Quercus robur L.) sont difficiles à stocker. En conséquence, la détérioration de leur qualité pendant le stockage peut réduire la production et la qualité des semis. Pour répondre à ceci, les effets de prétraitements de séchage et de trempage sur la germination des glands ont été évalués.• Après récolte à deux occasions en octobre, la teneur en eau (MC) des glands a été ajustée en utilisant les traitements suivants : (1) état frais ou témoin (46–48 %MC), (2) trempage à l’état frais (46–48 % MC), (3) séché (40–42 % MC) (méthode standard), (4) séché puis trempé (46–48 % MC). Les traitements permirent une germination des glands à 15 °C, aussi bien avant qu’après un stockage à −3 °C.• Les effets des traitements furent en général cohérents pour chaque date de récolte. Comparés aux traitements témoins, le trempage seul et le séchage suivi par un trempage ont accru significativement la germination aussi bien avant qu’après stockage. Le stockage réduit la germination sauf dans le cas des glands séchés et trempés.• Les résultats confirment que le séchage et le trempage améliorent la germination des glands, mais ceci n’est pas obtenu par l’intermédiaire d’un effet sur la teneur en eau des glands. Dans une expérimentation séparée en pépinière, ces traitements entraînent une plus grande production de semis et une meilleure qualité des plants que ceux issus du traitement standard (séché à 40–42 % MC.

Target seed moisture content, chilling and priming pretreatments influence germination temperature response in Alnus glutinosa and Betula pubescens

Abstract The response of common alder [Alnus glutinosa (L.) Gaertn.] and downy birch (Betula pubescens Ehrh.) seeds to germination temperature was examined following chilling and priming. Seeds of two seed lots of each species were subjected to combinations of chilling (4±1°C) and priming (20°C) treatments in fully imbibed (FI) state or a lower target seed moisture content (TMC) level (30% and 35% in alder and birch, respectively). After treatment, the seeds were allowed to germinate for 56 days at constant temperatures of 7.5, 10, 15, 20, 25 or 30°C. The response to temperature and pretreatment differed between species, but the effect was consistent in each seed lot within each species. In alder, the TMC seeds germinated well across the full range of temperatures, whereas there was an optimum temperature (22–23°C) for seeds given the FI pretreatment. Priming had no significant effect on the germination response of the TMC seeds in alder, but priming greatly improved germination in the FI seeds, especially at the lower germination temperatures (optimum 18–19°C). In contrast, in birch, the TMC seeds germinated better across the full temperature range, but the optimal germination temperature (15°C) was the same for all seed pretreatments. Priming improved germination in both the FI and TMC seeds in birch.

Effect of warm storage and date of lifting on the quality of Douglas-fir seedlings.

The effect of warm storage (15 °C)for up to 21 days on the vitality of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings lifted on four occasions from October to January wasassessed using fine root electrolyte leakage (REL). After 0, 7 and 21 days warm storage, additionalseedlings were potted and placed in a controlledgreenhouse environment to determine root growthpotential (RGP) after 4 weeks and survival after 8weeks. REL was a poor indicator of survival potential;REL increased after 7 days storage, despite the factthat RGP and survival were unaffected. REL increased further during the remainder of the 21-days storage period, but values were often not greatly different from those following 7days storage. Storage for up to 21 days greatlydecreased RGP and survival. Seedlings lifted inOctober and November were most affected by storagetreatments, but differences among lifting dates wererelatively small.

Germination response of alder and birch seeds to applied gibberellic acid and priming treatments in combination with chilling

The effects of seed moisture content (MC), gibberellic acid (GA3) concentration, chilling and priming pretreatments on the germination of common alder (Alnus glutinosa) and downey birch (Betula pubescens) seeds were examined. After treatment, the seeds were allowed to germinate for 42 days at 15 °C or 20 °C (dark)/30 °C (light). Treatment responses were similar at both temperatures and in both species. GA3 treatment of seeds in fully imbibed (FI) state for 30 days, or at the lower, target moisture content (TMC) for 30–90 days, significantly improved germination, but longer treatment periods reduced it (FI seeds) or had no effect (TMC seeds). Priming for two days improved germination in the FI seeds, but more than 4 days reduced it. Priming for up to 14 days had little effect on the germination of the TMC seeds.RésuméLes effets de l’humidité des graines (MC), de la concentration en acide gibbérellique (GA3), du froid et des prétraitements d’amorçage sur la germination des graines d:’Alnus glutinosa et de Betulus pubescens ont été examinés. Après traitement, les graines ont été mise à germer pour 42 jours à 15 °C ou 20 °C (a l’obscurité)/30 °C (à la lumière). Les réponses au traitement ont été similaires aux deux températures et pour les deux espèces. Le traitement, par GA3, des graines totalement imbibées (FI) pour 30 jours, ou de façon moindre, l’objectif d’humidité (TMC) pour 30–90 jours, ont amélioré significativement la germination, mais des périodes de traitements plus longues l’ont diminuée (graines FI) ou n’ont pas eu d’effet (graines TMC). Un amorçage de deux jours améliore la germination pour les graines FI, mais un amorçage de plus de 4 jours la diminue. Un amorçage jusqu’à 14 jours a eu peu d’effet sur la germination des graines TMC.

Impact of genetic variation and long-term limited water availability on the ecophysiology of young Sitka spruce (Picea sitchensis (Bong.) Carr.)

Future limited water availability may reduce the potential of tree improvement to increase timber yields. We investigated ecophysiological variation between full-sibling families of Sitka spruce (Picea sitchensis (Bong.) Carr.) growing under contrasting water availability conditions: control (optimal) water availability and limited water availability. One-year-old seedlings of nine improved families plus an unimproved seed lot were grown in pots in a greenhouse and the two water availability treatments imposed via drip irrigation. Whole-plant water use varied between families. Stomatal conductance and the light-saturated quantum yield of photosystem II at times differed between families, but not consistently. Certain families showed considerably greater increases in electron transport rate with increasing photosynthetically active radiation. Limited water availability resulted in reduced branch water potential, leaf stomatal conductance and transpiration per unit leaf area, and increased whole-plant water-use efficiency, in all genetic material. The responses of plant water use and leaf carbon isotope composition to water limitation, were, however, initially influenced by variation in vigour between families-with conservative growth in some material slowing the decline in substrate moisture content. As the duration of water deficit extended, these variables showed a more uniform response across families. Between-family variation in physiological mechanisms of drought tolerance was not detected. Thus, for Sitka spruce, assessing juvenile material may not allow selection to prevent reductions in productivity associated with long-term sub-optimal growing conditions, but screening for conservative growth (within families as well as between families) may be beneficial where survival of relatively short-term water limitation is the primary concern.

Effect of harvest date, drying, short-term storage and freezing after chilling on the germination of rowan seeds

ABSTRACT The effects of seed pretreatment and harvest date on the germination of European rowan (Sorbus aucuparia L) seeds were examined. In one experiment, seeds were subjected to drying, storage, soaking, warm and chilling treatments after harvesting in mid- and late August. In another experiment, fully imbibed (FI) seeds were given warm treatment for six weeks and then various durations of chilling (4 ± 1°C) for up to 24 weeks. Thereafter, the seeds were adjusted to target moisture content of 35% and 30% or remained in the FI state and were then subjected to either subsequent chilling or freezing (−3°C) for up to 32 weeks. The treated seeds were allowed to germinate at a constant 15°C with 8 hours of lighting per day. Treatment effects were generally consistent for each harvest date. Seeds did not germinate unless they were chilled and germination rates were low unless the seeds were soaked. Warm treatment applied before chilling appeared to induce dormancy. The effect of storage varied with harvest date and storage treatment, with germination being highest for seeds harvested in late August. Mild freezing of ≥16 weeks duration following 16–24 weeks initial chilling resulted in >80% germination, more than could be achieved using chilling alone.

Effect of deployment-type on stem growth, biomass partitioning and crown characteristics of juvenile Sitka spruce clones

Competitive interactions in clonal forestry are not well understood and this needs to be addressed to develop better deployment strategies. Eight juvenile Sitka spruce (Picea sitchensis (Bong.) Carriére) clones were grown in monoclonal and clonal mixtures in a field experiment for three years to assess the effects of genetic diversity on shoot growth, aboveand below-ground biomass partitioning and crown characteristics. Shoot elongation was measured throughout the growing season, while diameter was measured twice annually in May and December. After the third year, crown silhouette area was estimated from digitised images for one ramet per plot and ramets were then destructively harvested. Deployment × clone interactions were observed for tree height and diameter with reductions observed in mixed plots. Mixed plots had significantly greater height and diameter heterogeneity and more asymmetrical competition than monoclonal plots. Results from this study demonstrate that stem growth can be significantly altered when clones are planted in multi-clonal mixtures but for most clones, deployment-type will not significantly reduce their productivity.