Effect of cold and drought stress on blueberry dehydrin accumulation

Summary

Dehydrins are a group of plant proteins which respond to any type of stress that causes dehydration at the cellular level, such as cold and drought stress. Previously, three dehydrins of 65, 60, and 14.kDa were identified as the predominant proteins present in cold acclimated blueberry (Vaccinium corymbosumLinn.) floral buds. Levels were shown to increase with cold acclimation and decrease with deacclimation and resumption of growth. In the present study, to determine if dehydrins are induced in other organs in response to low temperature treatment (48C) and in response to drought, accumulation of dehydrins was examined in leaves, stems, and roots of two cultivars and one wild selection (a V. corymbosum cultivar, a V. ashei Reade cultivar, and a V. darrowi Camp selection) of blueberry by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting. Cold treatment involved placing plants in a cold room maintained at 48C for five weeks; drought stress was imposed by withholding water from potted, greenhouse-grown plants for 34.d. Relative water content (RWC) of shoots was determined periodically throughout the drought treatment. Dehydrins accumulated with both cold and drought stress but their molecular masses varied depending upon blueberry species. Dehydrins accumulated to higher levels in stems and roots than in leaves with cold stress and to higher levels in stems than in either roots or leaves with drought stress. Furthermore, cold treatment combined with dark treatment induced higher levels of dehydrins than cold treatment combined with a 10.h light/14.h dark photoperiod, suggesting that dehydrins may be responsive to changes in photoperiod as well. In the cold-stress experiment, the level of dehydrin accumulation was correlated with expected level of plant cold hardiness in the three genotypes. In the drought stress experiment, dehydrins accumulated prior to significant changes in RWC, and dehydrin levels did not appear to be closely correlated with RWC either among or within genotypes.     

Cold tolerance and short day acclimation in perennial Gaura coccinea and G. drummondii

The use of related species to integrate important traits into cultivated crops is a common practice in plant breeding. Gaura coccinea and, potentially its derived species Gaura drummondii, could be donor species for introgressing cold tolerance into non-hardy G. lindheimeri. However, cold tolerance and acclimation has not been studied in these species, so protocols for determining these traits are required. The objectives of this study were to determine the relative cold tolerance of G. coccinea, G. drummondii and whether short day photoperiod is involved in cold acclimation. G. drummondii from Texas, USA and G. coccinea from Minnesota and Texas, USA were subjected to freezing tests using whole plant or electrolyte leakage after natural acclimation or non-acclimation conditions. Minnesota genotypes were able to withstand colder temperatures (−12 °C) than Texas genotypes (−9 °C). Acclimation capacity was determined for whole plant and electrolyte leakage assays using three different plant organs—stem, crown, and rhizome. Underground rhizomes were the best predictors of cold tolerance, however they were difficult to obtain. Stem sections of G. drummondii (Texas) and G. coccinea (Minnesota and Texas) were used to determine a shift in acclimation under short days. The Minnesota G. coccinea genotypes demonstrated greater cold tolerance after 3 weeks of short days while Texas G. coccinea, G. drummondii genotypes did not change even after 5 weeks.     

Flushing and Leaf Growth of Cacao Under Controlled Temperature Conditions

Young clonal cacao trees have been grown for nine months in controlled- environment rooms at 74° F.,80°F. or 86° F. (23·3° C., 26·7° C., or 30·0° C.), or at one of these temperatures during the day and another during the night.

No specific temperature requirements for leaf flushing, which occurred in all the treatments, were found. Flushing was considerably greater at the higher temperatures, partly as a result of the loss of apical dominance, and was especially sensitive to day temperature.

The number of expanded leaves per flush and mean area per leaf increased with a decrease in day or night temperature, as did the duration of the leaves on the trees. The biggest net leaf area increases were made by plants given a day at 80° F. and a night at either 80° F. or 74° F. A night temperature of 86° F. resulted in a rapid turn-over of small unhealthy leaves having a low chlorophyll content.

The gain in total plant dry weight was greatest at a night temperature of 74° F., smallest at a night temperature of 86° F., and the plants which maintained the greatest leaf areas also gained the most dry weight. Leaf dry weight per unit leaf area was greatest at the lowest temperatures, as was the ratio of total plant dry weight to leaf area. The ratio of leaf weight to plant weight showed a small but significant increase with increase in day temperature.     

Growth of Rhododendron cultivars as affected by temperature and light

Summary

The purpose of this study was to investigate the adaptation of four Rhododendron cultivars to contrasting light and temperature conditions. Two evergreen rhododendron cultivars and two deciduous azaleas were grown for 112 d under short day (14 h) and long day (20 h) photoperiods combined with temperatures of 15 and 24°C. Additionally, these cultivars were compared for daylength extension at 24°C/long day under two irradiation treatments (incandescent lamps and fluorescent tubular lamps). The number of flushes of growth increased with increasing photoperiod and temperature in both evergreen cultivars and in R. canadense; azalea #89132 made only one flush in all treatments. In the evergreen cultivars the number of leaves per shoot in the first flush did not differ significantly between treatments, indicating that this character was predetermined by conditions during bud development. The number of leaves in later flushes increased with increasing photoperiod and temperature. The elongation growth of most flushes was also enhanced by longer photoperiod and higher temperature. High irradiation during photoperiodic extension further enhanced the growth. Azalea #89132 made more flower buds under high than low irradiation. The two evergreen cultivars differed in their growth habit. ‘Pohjola’s Daughter’ tended to continue growth in long days or at very high temperatures, and is thus predicted to thrive best in a maritime or semi-maritime cool climate. ‘Helsinki University’ responded to short daylength by ceasing growth regardless of temperature, and could be expected to perform successfully also in continental climates at latitudes around 45° N. R. canadense seemed to do best in a cool climate, but azalea #89132 should in time acclimatize in all kinds of climates within the limits of this study.     

Protocol: Streamline cloning of genes into binary vectors in Agrobacterium via the Gateway® TOPO vector system

Background

Freezing tolerance is an important factor in the geographical distribution of plants and strongly influences crop yield. Many plants increase their freezing tolerance during exposure to low, nonfreezing temperatures in a process termed cold acclimation. There is considerable natural variation in the cold acclimation capacity of Arabidopsis that has been used to study the molecular basis of this trait. Accurate methods for the quantitation of freezing damage in leaves that include spatial information about the distribution of damage and the possibility to screen large populations of plants are necessary, but currently not available. In addition, currently used standard methods such as electrolyte leakage assays are very laborious and therefore not easily applicable for large-scale screening purposes.

Results

We have performed freezing experiments with the Arabidopsis accessions C24 and Tenela, which differ strongly in their freezing tolerance, both before and after cold acclimation. Freezing tolerance of detached leaves was investigated using the well established electrolyte leakage assay as a reference. Chlorophyll fluorescence imaging was used as an alternative method that provides spatial resolution of freezing damage over the leaf area. With both methods, LT₅₀ values (i.e. temperature where 50% damage occurred) could be derived as quantitative measures of leaf freezing tolerance. Both methods revealed the expected differences between acclimated and nonacclimated plants and between the two accessions and LT₅₀ values were tightly correlated. However, electrolyte leakage assays consistently yielded higher LT₅₀ values than chlorophyll fluorescence imaging. This was to a large part due to the incubation of leaves for electrolyte leakage measurements in distilled water, which apparently led to secondary damage, while this pre-incubation was not necessary for the chlorophyll fluorescence measurements.

Conclusion

Chlorophyll fluorescence imaging is an alternative method to accurately determine the freezing tolerance of leaves. It is quick and inexpensive and the system could potentially be used for large scale screening, allowing new approaches to elucidate the molecular basis of plant freezing tolerance.     

Cold treatment modifies the photoperiodic flowering response of Lobelia×speciosa

Lobelia×speciosa Sweet ‘Compliment Scarlet' was grown under a range of photoperiods and low temperature treatments to determine their effects on flowering. In the first experiment, plants were held at 5°C for 0 or 15 weeks, then grown at 20°C under the following photoperiods: 10, 12, 14, 16, or 24 h of continual light or 9 h with a 4 h night interruption (NI). Non-cooled ‘Compliment Scarlet' flowered as a qualitative long-day plant (LDP) with a minimum flowering photoperiod of 14 h. Following cold, flowering was quantitative with respect to photoperiod, until ≈14.2 h, when the calculated rate of progress toward flowering reached a plateau. In cooled plants, node number below the inflorescence decreased from 27 to 16 as the photoperiod increased from 10 to 24 h. Cooled plants developed 61–149% more flowers and were ≥17% taller than non-cooled ones under the same photoperiod. To determine the cold duration required for flowering under short days (SD), plants were held at 0, 3, 6, 9, 12, or 15 weeks at 5°C then grown at 20°C under SD (9 h photoperiod) or long days (9 h photoperiod with a 4 h NI). Under SD, few plants flowered after ≤6 weeks of cold. As cold treatment increased from 9 to 15 weeks, flowering percentage increased, time to flower decreased from 93 to 64 days, and node count decreased from 24 to 13. Cold treatment did not affect flowering percentage or time under NI, but plants always had more flowers and were taller than reproductive ones under 9 h day lengths. Thus, ‘Compliment Scarlet', is a qualitative LDP, but an extended cold treatment can partially substitute for the long day (LD) photoperiodic requirement.     

Manipulation of bolting and flowering in celery (Apium graveolens L. var. dulce). III. Effects of photoperiod and irradiance

SUMMARY

Photoperiods of 8 and 16 h during chilling at 5°C had no effect on bolting and macroscopic flower appearance in celery cv. New Dwarf White. Eight hour photoperiods during chilling however markedly increased the number of plants forming sessile flowers. Short photoperiods (8 h) after chilling decreased the proportion of young, but competent plants that bolted and flowered. Total darkness during chilling completely prevented any subsequent vernalization response either as bolting or as flowering. Reducing irradiance receipt by the plants during chilling from 85 to W m"2 (PAR) had no effect on their vernalization response. After chilling, a reduction in mean daily total irradiance in the glasshouse from 4.05 to 1.57 MJ m"2 d-1 had no effect on bolting and flowering. Confinement of competent plants to darkness for 4-8 d at 20°C just prior to chilling resulted in a highly significant delay (F>0.001) to bolting and reduced the number of plants flowering. Two days of darkness had no significant effect. The inhibitory effects of dark treatments prior to chilling was greater in plants chilled subsequently for six weeks than for nine weeks.     

Changes in the contents of phenolic substances,phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) accompanying chilling-injury of eggplant fruit

The mechanism of browning, which is a typical chilling-injury of eggplant fruit (Solanum melongena L.), was investigated by determining the changes of phenolic substances, phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) either during storage at 1°C or after exposing fruit to low temperature for various periods.Chlorogenic acid and its isomer, the main substrates for browning, were isolated from eggplants by column and paper chromatography. R_f values and various color reactions of the above acids were compared with those of authentic chlorogenic acid.After 2 days of cold storage at 1°C, when browning was initiated, chlorogenic acid content decreased to less than half that of the initial day, rose to a maximum after 4 days, and then decreased rapidly.PAL activity increased to a peak after 2 days at 1°C, then decreased over 10 days as browning increased. TAL activity also increased after transfer from 1°C to 20°C.It is suggested that rapid turn-over of chlorogenic acid occurs in the early stage of cold storage of eggplant fruit, and development of browning is closely related to chlorogenic acid, PAL and TAL.     

Flower formation of Chinese cabbage. I. Response to vernalization and photoperiods

The influence of temperature, vernalization duration and photoperiods on the leaf number and beginning of bolting of Chinese cabbage was tested in growth chambers. An increasing effect of vernalization was indicated by decreasing leaf number and premature bolting. Sensitiveness for vernalization started with germination and remained constant with increasing plant age. During their development on mother plants, seeds were not vernalized.Inductive temperature ranged between 0 and 20°C. The necessary vernalization duration was lowest at 5–8°C. For slight vernalization effects one week was sufficient; for complete vernalization about 3 weeks were needed. To achieve the same effect, the necessary vernalization duration increased considerably at temperatures below 5°C, but increased only slightly at temperatures above 8°C.Long days promoted bolting when the temperature rose after incomplete vernalization. In constant low or high temperature, no photoperiodic effect was perceptible.     

The effect of light,darkness and temperature on micropropagation of the pear rootstock BP10030

There was no effect of irradiance level on surviving percentages of shoot tip explants of the pear rootstock BP10030, but low irradiance stimulated the initial growth of the explant. Irradiance had a strong effect on shoot multiplication. With an increase in photosynthetic photon flux (PPF) from 10 to 80 μmol m^?2s^?1, shoot number and length and shoot fresh and dry weights increased. The greatest number of shoots and the longest ones were obtained with a 16 h photoperiod, while the highest fresh and dry weight of shoots were produced with a 24 h photoperiod. Rooting percentage and the number of roots were markedly promoted under 80 μmolm^?2s^?1 PPF. Photoperiods of 8, 16 and 24 h produced similar effects on rooting percentages and the numbers of roots. Four to seven days of darkness were the optimum for rooting. Rooting percentage and the number of roots increased with increased temperature during darkness between 5 and 25°C. A further increase in dark temperature up to 30°C reduced rooting percentage and root number.     

Effects of exposure of bulbs to smoke and ethylene on flowering of Narcissus tazetta cultivar ‘Grand Soleil d'Or’

Bulbs of ‘Soleil d'Or’, exposed to smoke generated from smouldering wood and fresh leaves for several hours on each of 4 consecutive days during storage, produced flowers earlier and at a higher rate, even when using bulbs which were too small to flower using normal methods. The smoked bulbs showed an earlier start of floral initiation and faster development. A temperature of 25°C was optimal for storage. Application of ethylene also gave similar promotive effects when repeated 4 times at 10 μl 1^?1 for 1–5 h per day. Longer exposure to ethylene or smoke was less effective or had no promotive effect.     

Some physiological changes in strawberry (Fragaria × ananassa ‘Camarosa’) plants under heat stress

Summary

The effects of heat injury induced by long exposures were evaluated in strawberry (Fragaria × ananassa ‘plants’) Camarosa in this study. Seedlings were grown in 14 × 12 cm pots using perlite for three weeks at 25/10°C day/night temperature, and watered daily by modified 1/3 Hoagland nutrient solution. Half of the plants were transferred to a growth chamber with a constant 25°C, 16/8 h (light/dark) photoperiod regime and 1200 lux light intensity for a week to acclimate the plants. Temperature was increased stepwise (5 K per 48 h) to 30, 35, 40°C and finally to 45°C. In addition to others, plants were transferred from the outside to the growth chamber, at each temperature step to impose a heat shock. Leaf relative water content (RWC, %), loss of turgidity, chlorophyll content (Spad value) and heat-stress tolerance (HTS; LT₅₀) were measured in control and stressed plants. Total soluble proteins and total DNA were extracted from the leaves following the above treatments using standard procedures and total protein contents were determined using a Bradford assay. In general, effects of gradual heat stress (GHS) and shock heat stress (SHS) on the variables studied were mostly significant, except for chlorophyll content, while the effect of temperatures was significant for all the variables. Interaction between the heat stress type and temperature treatments was not significant for leaf RWC, loss of turgidity and chlorophyll content. Data also indicated that total protein and DNA contents were changed significantly by heat stress types (GHS and SHS) and/or temperature treatments. The plants exposed to GHS exhibited a significant increase in HST compared with the plants exposed to SHS (LT₅₀ of 41.5°C and 39°C, respectively). Consequently, gradual heat stress increased HST in strawberry leaves. Increased HST may be associated with the accumulation of several heat-stable proteins in GHS plants.     

Seed germination of seven pepper cultivars at constant or alternating high temperatures

Summary

Seeds of seven pepper (Capsicum annuum L.) cultivars (Anaheim TMR 23, California Wonder 300, Coronado, Jalapeno M, Ma Belle, Mercury, and Yolo Wonder B) were germinated at constant day and night temperatures of 25,30,35 and 40°C or at alternating temperatures of 40/25,40/30 and 40/35°C for 14 days. Germination percentages and rates were similar at 25 and 30°C. Largest differences in cultivar responses occurred at 35°C where germination percentages ranged from 24 to 96%, and rates, calculated as summation of the number of seeds germinated on a given day divided by day number, varied from 3 to 26 (theoretical maximum value of 100). At 40°C, germination percentages were less than 5% and rates were less than one for all cultivars. Cultivars with the most heat tolerance were ‘Mercury’ and ‘Yolo Wonder B’. At alternating temperatures, germination percentages and rates were higher than those at constant 40°C. The increases were greatest when the temperature was lowered by 15°C (40/25°C) and least when temperatures were lowered by 5°C (40/35°C). Tetrazolium tests showed that a large percentage of the ungerminated seed was still viable from the highest temperature. At lower temperatures, fewer ungerminated seeds were viable with no viable ungerminated seeds from the lowest temperature.     

Flower formation in Calceolaria × herbeohybrida Voss

At high light intensity Calceolaria × herbeohybrida ‘Zwerg Meisterstück’ is a long-day plant with a critical day-length of 14–15 h. At low light intensity (e.g. as in winter) flowering will take place if the long-day treatment is preceded by a chilling period (10°C) or by short days at 15–20°C. During the chilling period day-length is of little influence. With increasing duration of the chilling period the requirement for long days decreases and the critical day-length becomes shorter. After a sufficient chilling period, flowering occurs both in long and short days. If the chilling period lasts approx. 40 days, however, flowering in short days is delayed, phyllody occurs and only a small number of flowers develop in comparison to long days. After at least 70–75 days of chilling plants show almost the same reaction in short and long days. Plants not chilled under conditions of high light intensity and short days flower either in naturally long days or by day extension with incandescent light. After chilling, fluorescent light of the type L 39 is also effective for day extension. A night break with incandescent light in a 16-h dark period induces flowering only after a chilling period. Incandescent but not fluorescent light causes a slight yellowing and more upright position of the leaves and an elongation of the internodes.     

优质耐寒白菜新品种‘海青’

‘海青’白菜为优质耐寒一代杂种。植株直立，中矮箕类型，生长势强；叶深绿色，叶面光滑，叶柄宽、绿色，品质佳，播种后25 ~ 40 d均可采收，40 d时采收平均产量可达40 500 kg • hm-2，耐寒性强，抗病毒病和霜霉病。适宜在上海及长江中下游类似生态条件地区秋冬季栽培。     

Promotion of floral initiation in ‘Fuerte’ avocado by low temperature and short daylength

Potted avocado (Persea americana Mill., cv. ‘Fuerte’) plants were maintained in growth cabinets for up to 32 weeks and new growth observed for flower formation. Flowers were formed if temperatures were 20°C or below, but with 25° or 30°, even if only for 1 hour per day, flower formation was inhibited. Time to flowering was accelerated, but number of flowers reduced, if daylength was shortened from 15 h to 9 h. With low temperature and short days, full bloom was about 4 months after starting experiments. Spring flowering of cv. ‘Fuerte’ in the field could follow flower induction about 4 months previously with the onset of winter temperatures and daylengths.     

Chilling induced ethylene biosynthesis in ‘Hayward’ kiwifruit following storage

‘Hayward’ kiwifruit were stored at 0, 5, 10, 15 and 20°C for 5, 12 and 17 days before rewarming to 20°C for 10 more days. Ethylene and CO₂ production, ACC, ACC synthase (ACS) and ACC oxidase (ACO) activities, flesh and core firmness, soluble solids content (SSC) and flesh colour were measured. Kiwifruit stored at 0, 5, 10 and 15°C did not ripen, produce ethylene or show increases in ACS or ACO activity. Fruit stored for 5 days at the above temperatures, then rewarmed to 20°C, did not show any change during the following 10 days. Rewarmed fruit, pre-stored at 0–10°C for 12 days, started autocatalytic ethylene production within 24 h, followed by fruit ripening. Fruit stored at 15°C for 12 days needed 72 h to start ethylene autocatalyse and did not fully ripen during 10 days at 20°C. After 17 days storage at 0–15°C kiwifruit started autocatalytic ethylene production with no delay upon exposure to 20°C. Autocatalytic ethylene production correlated with increased ACC content, and increased activities of ACS and ACO. Fruit held continuously at 20°C started autocatalytic ethylene production after 19 days, with concomitant increases in ACC content, ACS and ACO activities and ripening. Respiration increased after rewarming, concomitantly with the increase in ethylene production.We concluded that exposing kiwifruit to chilling temperatures (0–10°C) for 12 days advanced ethylene biosynthesis and ripening when compared with fruit held continuously at 20°C. The advanced ethylene biosynthesis was due to increase ACS and ACO activities immediately upon rewarming of the fruit.     

Influence of temperature on the vegetative and generative development of Haemanthus × Hybridus cv. ‘König Albert’

The influence of temperature on breaking dormancy, and on stimulating flower initiation and development has been studied.The largest number of foliage leaves per plant developed after a dormant period of 80–120 days at a temperature of 10–15°C. The greater the number of foliage leaves, the higher the probability of formation of an inflorescence. The cooling period at 10–15°C must follow immediately after the vegetation period.Plants grown continuously at 25°C developed few or no foliage leaves after dormancy and died after 2 or 3 years. Inflorescences, developed after a dormant period above 17°C, were of poor quality. The later the cooling period began after wilting of the foliage leaves, the greater number of young inflorescences died.Bulbs stored under unfavourable conditions differentiated fewer primordia, and initiation of inflorescences, with a few exceptions, did not occur. A dormancy period at 5°C or less is fatal to the plants.     

Cane Morphology and Anatomy Influence Freezing Tolerance in Vitis vinifera Cabernet franc

In 2014 and 2015, Ohio vineyards were exposed to multiple freeze events of –20 °C or lower, resulting in vine dieback, i.e., complete damage of above ground parts in Vitis vinifera. Grapevines that sustained dieback were rehabilitated for trunk replacement by training 1-year-old shoots with two distinct morphologies, based on internode diameter of large (L) and normal (N). This study evaluated the impact of cane morphology (L and N) in V. vinifera Cabernet franc on freezing tolerance (FT) of bud, phloem, and xylem tissues in relation to their respective anatomical structures and carbohydrate concentrations. Compared to N canes (7–9-mm diameter), L canes (10–15 mm) in Cabernet franc were considered vigorous and had the following morphological characteristics: long and heavy, with long and wide internode, and presence of numerous laterals. Furthermore, cane anatomy was also different with L canes having a significantly higher number of vascular transport units, xylem vessels, and phloem fibers than those in N canes. Freezing tolerance of buds and phloem was also different between the two cane types, with L canes being more cold sensitive than N canes, especially during fall acclimation and late-winter deacclimation. Sugar concentrations, however, were not different between L and N canes. These results suggest that cane morphology and anatomy play a significant role in affecting FT and the large and abundant anatomical structures of phloem and xylem contributed to the reduced FT of these tissues. In all practicality, this study suggests the best cultural practice for trunk replacement, and vine recovery should include the removal of the undesirable vigorous and cold sensitive canes during pruning.     

Temperature and development in Iris × hollandica during preplanting storage. I. Leaf initiation

Summary

Leaf initiation was examined in Dutch iris bulbs during pre-planting storage temperature treatments in the dark. The number of leaves initiated before inflorescence evocation increased with increasing temperature. The base, optimum and maximum temperature for leaf initiation were established as –0.4, 13.1 and 26.7°C respectively. The rate of leaf initiation was shown to be linearly related to temperature. The average thermal-time required for each leaf to be initiated under constant temperatures was 79°Cd but leaves initiated during the transfer temperature treatments required an average of 92°Cd. Rates of leaf initiation predicted from thermal-time equations were similar to those observed in bulbs stored at the lower temperatures (2–13°C) but rates observed at warmer temperature (17–25°C) never reached the predicted high value.