Evaluation of chilling requirements for six Arkansas blackberry cultivars utilizing stem cuttings

Woody perennial plants including blackberries (Rubus subgenus Rubus) require certain amounts of chilling or rest hours below 7ºC during the dormant season for successful bud break the following year. Arkansas-developed blackberry cultivars are being grown in various climates worldwide and all cultivars need chilling requirement estimates for accurate recommendations of adaptation. Determining chilling requirement using stem cuttings collected from field-grown plants rather than whole plants is a desirable system. We conducted a study to evaluate both artificial and field chilling of six cultivars. For the artificial-chilling study, 12- node stem cuttings were collected 2 days after the first killing frost. These were then placed in a moist medium in a walk-in cooler at 3ºC. At 100 hour chilling intervals, five cuttings of each cultivar were placed under an intermittent mist system. For the field-chilling study, a biophenometer was placed in the field to measure chill, and ten 12-node stem cuttings of each cultivar were collected at 100-hour intervals of chilling up to 1000 hours below 7ºC and placed under mist. For both studies the mist bench was located in a heated greenhouse (min. temperature of 15ºC), and cuttings were placed according to a completely random design. Budbreak was recorded weekly. Studies were analyzed separately by SAS. Results for Study One, artificial-chilling, were inconclusive due to a lack of clear differentiation among the cultivars and chilling intervals. Study Two, using field-chilling, showed a significant chilling interval x cultivar interaction. ‘Arapaho’ appeared to have a chilling requirement of 400 to 500 hours, ‘Kiowa’ 200 hours, ’Shawnee’ 400 to 500 hours, and ‘Chickasaw’ possibly 600 to 700 hours. The cultivars Choctaw and Apache did not provide clear chilling interval differentiation in the study. Our results indicate that the use of stem cuttings receiving field chilling to evaluate chilling requirement of blackberry cultivars has merit and can be a successful method in this research area

Tapping into the maize root microbiome to identify bacteria that promote growth under chilling conditions

Background When maize (Zea mays L.) is grown in the Northern hemisphere, its development is heavily arrested by chilling temperatures, especially at the juvenile phase. As some endophytes are beneficial for plants under stress conditions, we analyzed the impact of chilling temperatures on the root microbiome and examined whether microbiome-based analysis might help to identify bacterial strains that could promote growth under these temperatures. Results We investigated how the maize root microbiome composition changed by means of 16S rRNA gene amplicon sequencing when maize was grown at chilling temperatures in comparison to ambient temperatures by repeatedly cultivating maize in field soil. We identified 12 abundant and enriched bacterial families that colonize maize roots, consisting of bacteria recruited from the soil, whereas seed-derived endophytes were lowly represented. Chilling temperatures modified the root microbiome composition only slightly, but significantly. An enrichment of several chilling-responsive families was detected, of which the Comamonadaceae and the Pseudomonadaceae were the most abundant in the root endosphere of maize grown under chilling conditions, whereas only three were strongly depleted, among which the Streptomycetaceae. Additionally, a collection of bacterial strains isolated from maize roots was established and a selection was screened for growth-promoting effects on juvenile maize grown under chilling temperatures. Two promising strains that promoted maize growth under chilling conditions were identified that belonged to the root endophytic bacterial families, from which the relative abundance remained unchanged by variations in the growth temperature. Conclusions Our analyses indicate that chilling temperatures affect the bacterial community composition within the maize root endosphere. We further identified two bacterial strains that boost maize growth under chilling conditions. Their identity revealed that analyzing the chilling-responsive families did not help for their identification. As both strains belong to root endosphere enriched families, visualizing and comparing the bacterial diversity in these communities might still help to identify new PGPR strains. Additionally, a strain does not necessarely need to belong to a high abundant family in the root endosphere to provoke a growth-promoting effect in chilling conditions

Determination of Chilling Requirement of Arkansas Thornless Blackberry Cultivars

Little research has been done to determine the chilling requirement for blackberry cultivars. However, field observations from areas where fewer hours of chilling occur indicate that ‘Navaho’ requires more hours of chilling than does ‘Arapaho’. The objective of our study was to determine a method for measuring the chilling requirement using whole plants of two blackberry cultivars, Arapaho and Navaho. One-year old, bare-root plants were field-dug on 26 October 1999 and placed in a cold chamber at 3ºC. Ten single-plant replications of each cultivar were removed at 100-hour intervals up to 1000 hours. The plants were potted and placed in a greenhouse (daily minimum temperature 15ºC), and plants were arranged on benches in a completely randomized design. Budbreak was recorded on a weekly basis. Data for budbreak were analyzed as a two-factor factorial (2 cultivars and 10 chilling treatments) by SAS and means were separated by least significant difference (P = 0.05). Data indicated that the chilling requirement for Arapaho is between 400 and 500 hours. For Navaho, the data indicated the chilling requirement was between 800 and 900 hours. These data support previous observations and indicate that the method used was successful in determining the chilling requirement for blackberries

Assessment of Five Chilling Tolerance Traits and GWAS Mapping in Rice Using the USDA Mini-Core Collection

Rice (Oryza sativa L.) is often exposed to cool temperatures during spring planting in temperate climates. A better understanding of genetic pathways regulating chilling tolerance will enable breeders to develop varieties with improved tolerance during germination and young seedling stages. To dissect chilling tolerance, five assays were developed; one assay for the germination stage, one assay for the germination and seedling stage, and three for the seedling stage. Based on these assays, five chilling tolerance indices were calculated and assessed using 202 O. sativa accessions from the Rice Mini-Core (RMC) collection. Significant differences between RMC accessions made the five indices suitable for genome-wide association study (GWAS) based quantitative trait loci (QTL) mapping. For young seedling stage indices, japonica and indica subspecies clustered into chilling tolerant and chilling sensitive accessions, respectively, while both subspecies had similar low temperature germinability distributions. Indica subspecies were shown to have chilling acclimation potential. GWAS mapping uncovered 48 QTL at 39 chromosome regions distributed across all 12 rice chromosomes. Interestingly, there was no overlap between the germination and seedling stage QTL. Also, 18 QTL and 32 QTL were in regions discovered in previously reported bi-parental and GWAS based QTL mapping studies, respectively. Two novel low temperature seedling survivability (LTSS)–QTL, qLTSS3-4 and qLTSS4-1, were not in a previously reported QTL region. QTL with strong effect alleles identified in this study will be useful for marker assisted breeding efforts to improve chilling tolerance in rice cultivars and enhance gene discovery for chilling tolerance

Indicator organisms to determine the use of chilling as a critical point in beef slaughter HACCP

End of project reportDuring chilling, temperatures of carcass surfaces at different sites change over time as do other parameters such as water activity (aw), the structure of the muscle and other tissues, as the carcass enters rigor mortis. Many of these factors are known to have a major effect on cell survival and growth and must be considered in determining the influence of chilling on bacterial survival on carcass surfaces. This study aimed to determine if chilling could be used as a critical control point (CCP) in beef slaughter in relation to pathogens such as E. coli O157:H7 and L. monocytogenes, using E. coli and Listeria innocua as pathogen indicators. The present study was designed to determine the influence of (a) chilling at 10oC for 72 h on the survival of E. coli and (b) chilling at 4oC for 72 h on the survival of L. innocua inoculated at different sites on beef carcasses. Three sites (neck, outside round and brisket) were inoculated (1) immediately after dressing while hot (E. coli and L. innocua) and (2) when cold after chilling (L. innocua). The influence of changes in surface aw was also considered and their relationship to the survival of E. coli and L. innocua over time was assessed. The data are discussed in relation to the use of chilling as a CCP in beef hazard analysis (HACCP) and the monitoring of neck temperature as the most suitable CCP.National Development Pla

Effects of chilling on the expression of ethylene biosynthetic genes in Passe-Crassane pear (Pyrus communis L.) fruits

Passe-Crassane pears require a 3-month chilling treatment at 0 C to be able to produce ethylene and ripen autonomously after subsequent rewarming. The chilling treatment strongly stimulated ACC oxidase activity, and to a lesser extent ACC synthase activity. At the same time, the levels of mRNAs hybridizing to ACC synthase and ACC oxidase probes increased dramatically. Fruit stored at 18 C immediately after harvest did not exhibit any of these changes, while fruit that had been previously chilled exhibited a burst of ethylene production associated with high activity of ACC oxidase and ACC synthase upon rewarming. ACC oxidase mRNA strongly accumulated in rewarmed fruits, while ACC synthase mRNA level decreased. The chilling-induced accumulation of ACC synthase and ACC oxidase transcripts was strongly reduced when ethylene action was blocked during chilling with 1-methylcyclopropene (1-MCP). Upon rewarming ACC synthase and ACC oxidase transcripts rapidly disappeared in 1-MCP-treated fruits. A five-week treatment of non-chilled fruits with the ethylene analog propylene led to increased expression of ACC oxidase and to ripening. However, ethylene synthesis, ACC synthase activity and ACC synthasemRNAs remained at very lowlevel. Our data indicate thatACC synthase gene expression is regulated by ethylene only during, or after chilling treatment, while ACC oxidase gene expression can be induced separately by either chilling or ethylene

Oxidative Stress Associated with Chilling Injury in Immature Fruit: Postharvest Technological and Biotechnological Solutions

Immature, vegetable-like fruits are produced by crops of great economic importance, including cucumbers, zucchini, eggplants and bell peppers, among others. Because of their high respiration rates, associated with high rates of dehydration and metabolism, and their susceptibility to chilling injury (CI), vegetable fruits are highly perishable commodities, requiring particular storage conditions to avoid postharvest losses. This review focuses on the oxidative stress that affects the postharvest quality of vegetable fruits under chilling storage. We define the physiological and biochemical factors that are associated with the oxidative stress and the development of CI symptoms in these commodities, and discuss the different physical, chemical and biotechnological approaches that have been proposed to reduce oxidative stress while enhancing the chilling tolerance of vegetable fruits

Delayed chilling appears to counteract flowering advances of apricot in southern UK

Temperatures are rising across the globe, and the UK is no exception. Spring phenology of perennial fruit crops is to a large extent determined by temperature during effective chilling (endo-dormancy) and heat accumulation (eco-dormancy) periods. We used the apricot flowering records of the UK National Fruit Collections (NFC) to determine the influence of temperature trends over recent decades (1960 to 2014) on apricot (Prunus armeniaca L.) flowering time. Using Partial Least Squares (PLS) regression, we determined the respective periods for calculating chill and heat accumulation. Results suggested intervals between September 27th and February 26th and between December 31st and April 12th as the effective chilling and warming periods, respectively. Flowering time was correlated with temperature during both periods, with warming during chilling corresponding to flowering delays by 4.82 d°C-1, while warming during heat accumulation was associated with bloom advances by 9.85 d°C-1. Heat accumulation started after accumulating 62.7 ± 5.6 Chill Portions, and flowering occurred after a further 3744 ± 1538 Growing Degree Hours (above a base temperature of 4°C, with optimal growth at 26°C). When examining the time series, the increase in temperature during the chilling period did not appear to decrease overall chill accumulation during the chilling period but to delay the onset of chill accumulation and the completion of the the average chill accumulation necessary to start heat accumulation. The resulting delay in heat responsiveness appeared to weaken the phenology-advancing effect of spring warming. These processes may explain why apricot flowering time remained relatively unchanged despite significant temperature increases. A consequence of this may be a reduction of frost risk for early flowering crops such as apricot in the UK

Development of a mathematical model for 'Hayward' kiwifruit softening in the supply chain : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Food Technology at Massey University, New Zealand

Fruit loss is a major concern to the kiwifruit industry as it incurs a high cost to monitor and remove over soft or rotten fruit to meet export standards. Kiwifruit is exposed to various temperature scenarios due to different packhouse cooling practices, and temperature control is difficult to maintain throughout the supply chain. Fruit pallet temperatures are wirelessly monitored in the supply chain. This time temperature data provides valuable rich information which could be used to predict kiwifruit quality. In the laboratory, green ‘Hayward’ kiwifruit were exposed to industry coolchain scenarios to investigate their influence on fruit firmness in subsequent storage. Cooling rate and storage temperature were identified to affect fruit firmness and chilling injury development significantly, where accelerated softening and increased chilling injury development was observed in late storage (> 100 d) when fruit were cooled directly to 0 °C. However, when fast cooled fruit were stored at 2 °C instead of 0 °C, low incidence of chilling injury was observed. The influence of cooling rate and storage temperature on kiwifruit quality suggests that industry should focus on the management practices adopted by packhouses in order to maintain acceptable quality after long term storage. A proportion of the firmness data results were used to develop a mechanistic style mathematical model of kiwifruit softening. Kiwifruit softening was mathematically described based on the correlation with starch degradation, breakdown of cell wall structure, and a description of the incidence of chilling injury development during storage. The model inputs consist of solely commonly collected at-harvest attributes: firmness, dry matter and soluble solids content and time-temperature data. Applying at-harvest attributes as model inputs enabled a capability to predict different softening curves as influenced by fruit maturity, and grower line differences. The developed model demonstrated promising softening prediction with mean absolute errors (MAE) between 0.8 to 2.1 N when fruit were exposed to fluctuating temperatures and cooling profiles. A logistic model was used to estimate the proportion of chilling injured fruit. Based on the given time temperature information, the logistic model was able to predict the proportion of chilling injured fruit reasonably well (R2 = 0.735). This chilling injury prediction was subsequently used to adjust the softening prediction during the late storage period (>100 d). Model validation was performed using the remaining data, identifying a lack of fit in both the rapid (MAE of 20.8 N) and gradual (MAE of 8.0 N) softening phase. The lack of fit in the rapid softening phase is proposed to be explained by the presence of an initial lag phase in softening which the developed model is unable to predict. The magnitude of firmness associated with starch content and cell wall integrity heavily influenced the lack of fit in the gradual softening phase. Fixing the initial amount of firmness associated to cell wall integrity to be constant for all maturities and grower lines improved the softening prediction. Overall, this thesis contributes to the challenge of predictively modelling kiwifruit quality in the supply chain. However, there are still many opportunities for improvement including introducing the influence of: variation within the same batch; fruit maturity on chilling injury development; ethylene in the environment; pre-harvest management practices and extending the model to have more focus on high temperature conditions such as those experienced in the marketplace. Conducting studies on: the effect of curing on kiwifruit; using non-destructive techniques to provide information to help define model parameters for prediction; effect of high temperature exposure on kiwifruit softening are possible opportunities that may contribute to enable better prediction of kiwifruit quality in the supply chain in the future

The capacity to maintain ion and water homeostasis underlies interspecific variation in Drosophila cold tolerance

Many insects, including Drosophila, succumb to the physiological effects of chilling at temperatures well above those causing freezing. Low temperature causes a loss of extracellular ion and water homeostasis in such insects, and chill injuries accumulate. Using an integrative and comparative approach, we examined the role of ion and water balance in insect chilling susceptibility/ tolerance. The Malpighian tubules (MT), of chill susceptible Drosophila species lost [Na+] and [K+] selectivity at low temperatures, which contributed to a loss of Na+ and water balance and a deleterious increase in extracellular [K+]. By contrast, the tubules of chill tolerant Drosophila species maintained their MT ion selectivity, maintained stable extracellular ion concentrations, and thereby avoided injury. The most tolerant species were able to modulate ion balance while in a cold-induced coma and this ongoing physiological acclimation process allowed some individuals of the tolerant species to recover from chill coma during low temperature exposure. Accordingly, differences in the ability to maintain homeostatic control of water and ion balance at low temperature may explain large parts of the wide intra- and interspecific variation in insect chilling tolerance