Effect of 1,2-benzisoxazole-3-acetic acid on adventitious shoot regeneration and in vitro rooting in apple

 The effect of 1,2-benzisoxazole-3-acetic acid (BOA), compared to 1-naphthaleneacetic acid (NAA), on adventitious shoot formation in leaf portions and compared to indolebutyric acid (IBA), on in vitro rooting in the apple (Malus domestica Borkh) cultivars McIntosh and Gala, and one rootstock, Jork 9, was investigated. BOA at 43.0 μm or 2.7 μm at NAA in combination with 17.8 μm benzyladenine (BA), induced the highest number of explants to produce adventitious shoots in Jork 9. In Gala, the combination of 21.5 μm BOA with 1.0 μm thidiazuron (TDZ) or with 22.0 μm BA induced the highest regeneration percentages, 58 and 54%, respectively, giving more satisfactory results than NAA (where only 42% of leaf explants exhibited shoot formation). In McIntosh, the highest percentage of regeneration was obtained with 1.3 μm NAA and 22.0 μm BA, while 51% was the highest response obtained with the BOA treatment. The combination of BOA with TDZ completely inhibited regeneration activity in leaf portions of this cultivar. The shoots of all the genotypes obtained with the most morphogenetic NAA or BOA treatments were excised, multiplied and successfully rooted and hardened. The results demonstrate that the synthetic auxin BOA is active in inducing shoot regeneration from leaf explants of apple and that the activity of BOA in plant regeneration is genotype dependent. When BOA was used to induce rooting in apple microcuttings, lower rooting percentages were obtained than with IBA, showing that the effect of BOA in inducing root formation is very low and that it cannot be used routinely to replace IBA in the in vitro rooting of microcuttings. Received: 18 June 1998 / Revision received: 4 January 1999 / Accepted: 29 January 1999     

Responses ofHevea brasiliensis Muell. Arg. plants to low temperatures under controlled conditions

In this study we have examined the responses ofHevea plants to low temperatures in growth chambers simulating winter conditions in the tropics. The low temperatures increased the leaf diffusive resistance even with a rise in the leaf temperatures toward the end of day. The total chlorophyll levels were reduced by chilling, with no changes in the chlorophyll a/b ratio. Neither organic solute nor electrolyte leakage were observed at low temperatures. These results explain the low growth rate of plants and the lack of serious chilling symptoms (necrosis) in mature leaves during winter conditions near latitude 22° S in Brazil.     

Influence of soil drying on root development,water relations and leaf growth of Ceratonia siliqua L.

Summary Seedlings of Ceratonia siliqua L., an evergreen sclerophyll species native to the Mediterranean region, were grown in 30-cm deep tubes of John Innes II potting compost in a growth cabinet maintained at 15° C during a 12-h day where PAR was 400 mol m^–2 s^–1. After a period of acclimatisation to the conditions in the cabinet during which plants were watered every day, water was withheld from the soil in some tubes for 24 days. These conditions may be regarded as a simulation of the natural situation. Estimates of leaf and root water potential and solute potential, leaf growth and root development were made at intervals during the soil drying cycle on both watered and unwatered plants. Water potential and solute potential measurements were made both on young expanding and on fully expanded leaves. During the experimental period, root growth of C. siliqua was not much affected by soil drying, and roots in both the watered and the unwatered columns penetrated to the bottom of the soil tubes by the end of the drying treatment. Expanded leaves showed significant limitation in stomatal conductance as soil drying progressed. Leaf water potential of fully expanded leaves of unwatered plants declined substantially. In contrast, water potential of young expanding leaves on unwatered plants declined to only a limited extent and turgor was sustained. As the soil dried, stomatal conductance of young leaves was always higher than that of mature leaves; also, placticity and elasticity of young leaves slowly decreased whereas mature leaves became stiff. Changing leaf cell wall properties may determine different patterns of water use as the leaves age. A mechanism of continuous diffusion of water through the soil towards the tip and pumping towards the young leaves is proposed.     

Influence of root temperature on growth of Pinus sylvestris,Fagus sylvatica,Tilia cordata and Quercus robur

One-year-old tree seedlings were incubated in a greenhouse from April to July, under natural daylight conditions, with their root systems at constant temperatures of 5, 10, 15, 20, 25, 30 and 35 °C and with the above ground parts kept at a constant air temperature of 18–20 °C. The course of height growth, total mass increment, root, shoot and leaf weight as well as leaf areas were measured. The results indicate that clear differences exist in the optimal root zone temperatures for various growth parameters in different tree species. Pinus sylvestris had a maximal height increment at about 5–10 °C and maximal total mass increment at 15 °C root temperature. In contrast, the optimum for Quercus robur was at 25 °C. Tilia cordata and Fagus sylvatica had their optima for most growth parameters at 20 °C. The root temperature apparently indirectly influenced photosynthesis (dry weight accumulation) and respiration loss. From the observed symptoms and indications in the literature it seems probable that a change in hormone levels is involved as the main factor in the described effects. Variation of root temperature had only an insignificant effect on bud burst and the time at which the shoots sprouted. Apparently species of northern origin seem to have lower root temperature optima than those of more southern origin. This is to be verified by investigation of other tree species.     

Physiological Responses of Apple Trees to Supraoptimal Root Temperature

Ungrafted apple rootstocks were grown in sand cultures at constant root temperatures between 20°C to 40°C. Temperatures of 30°C and above reduced root and shoot growth. Serious damage to the leaves occurred at 35°C and above. The O₂ consumption, CO₂ evolution and respiratory quotient (RQ) of the roots showed maximum values at 35°C. Different rootstock cultivars varied greatly in their susceptibility to damage by supraoptimal root temperatures apparently due to anaerobic respiration. The more susceptible ones differed from resistant types in the larger amount of ethanol they accumulated in their roots at supraoptimal root temperature, and the more severe reduction in the malic acid content of the roots at such temperature. Acetaldehyde was also found in roots and leaves at supraoptimal root temperatures, whereas the organic acid content of the leaves tended to decrease. Supraoptimal root temperature also caused a reduction of cytokinins in both roots and leaves accompanied by a reduction in the leaf chlorophyll content. This could be prevented by the application of kinetin or benzyladenine to the leaves. In a short experiment a rise in root temperature up to 40°C caused an increase in transpiration and a decrease in the resistance of the leaves to the passage of water vapor, whereas in prolonged experiments transpiration reached a maximum and leaf resistance a minimum at 30°C. The leaf water potential increased also with increasing root temperature. Leaf temperature increased with increasing root temperature, irrespective of increasing or decreasing transpiration rates.     

Growth of Escherichia coli O157:H7 in Bruised Apple (Malus domestica) Tissue as Influenced by Cultivar, Date of Harvest, and Source

Four of five apple cultivars (Golden Delicious, Red Delicious, McIntosh, Macoun, and Melrose) inoculated with Escherichia coli O157:H7 promoted growth of the bacterium in bruised tissue independent of the date of harvest (i.e., degree of apple ripening) or the source of the apple (i.e., tree-picked or dropped fruit). Apple harvest for this study began 4 September 1998 and ended 9 October, with weekly sampling. Throughout this study, freshly picked (<2 days after harvest) McIntosh apples usually prevented the growth of E. coli O157:H7 for 2 days. Growth of E. coli O157:H7 did occur following 6 days of incubation in bruised McIntosh apple tissue. However, the maximum total cell number was approximately 80-fold less than the maximum total cell number recovered from Red Delicious apples. When fruit was stored for 1 month at 4°C prior to inoculation with E. coli O157:H7, all five cultivars supported growth of the bacterium. For each apple cultivar, the pH of bruised tissue was significantly higher and °Brix was significantly lower than the pH and °Brix of undamaged tissue regardless of the source. In freshly picked apples, changes in the pH did not occur over the harvest season. Bruised Golden Delicious, McIntosh, and Melrose apple tissue pHs were not significantly different (tree-picked or dropped), and the °Brix values of McIntosh, Macoun, and Melrose apple tissue were not significantly different. Single-cultivar preparations of cider did not support growth of E. coli, and the cell concentration of inoculated cider declined over an 11-day test period. The rate of decline in E. coli cell concentration in the McIntosh cider was greater than those in the other ciders tested. The findings of this study suggested that the presence of some factor besides, or in addition to, pH inhibited E. coli growth in McIntosh apples.     

Leaf senescence and grain filling affected by post-anthesis high temperatures in two different wheat cultivars

High temperature is a major factor affecting grain yield and plant senescence in wheat growing regions of central and east China. In this study, two different wheat cultivars, Yangmai 9 with low-grain protein concentration and Xuzhou 26 with high-grain protein concentration, were exposed to different temperature regimes in growth chambers during grain filling. Four day/night temperature regimes of 34°C/22°C, 32°C/24°C, 26°C/14°C, and 24°C/16°C were established to obtain two daily temperatures of 28 and 20°C, and two diurnal day/night temperature differences of 12 and 8°C. Concentration of a lipid peroxidation product malondialdehyde (MDA), activities of the antioxidants superoxide dismutase (SOD) and catalase (CAT), chlorophyll concentration (SPAD) in flag leaves and kernel weight were determined. Results show that activities of SOD and CAT in leaves increased markedly on 14 days after anthesis (DAA) for the high-temperature treatment (34°C/22°C) and then declined. As a result, MDA concentration in leaves increased significantly under high temperature (34°C/22°C and 32°C/24°C). Compared with optimum temperature treatment, high temperature reduced the concentration of soluble protein and SPAD values in flag leaves. Grain-filling rate increased slightly initially, but decreased significantly during late grain filling under high temperature. As a result, final grain weight was reduced markedly under high temperature. Decreases in the activities of SOD and CAT and increases in MDA concentration in leaves were more pronounced with a 12°C of day/night temperature difference when under high temperatures. Kernel weight was higher under 12°C of day/night temperature difference under optimum temperatures (24°C/16°C and 26°C/14°C). The responses to high-temperature regimes appeared to differ between the two wheat cultivars with different grain protein concentrations. It is concluded that a larger diurnal temperature difference hastened the senescence of flag leaves under high-temperature conditions, but retarded senescence under optimum temperature treatments of 26°C/14°C and 24°C/16°C.     

Effects of growth temperature and winter duration on leaf phenology of Erythronium japonicum,a forest spring geophyte

The effects of growth temperature and winter duration on the leaf phenology of Erythronium japonicum were examined in two experiments. Bulbs wintered in the field were cultivated at 10 and 20° C and the bulbs were cultivated at 15° C after chilling treatment at 3° C for 60 and 120 days and without chilling in winter. The plants cultivated at 20° C showed significantly earlier leaf emergence, a more rapid rate of leaf extension and shorter leaf longevity than those cultivated at 10° C. The decrease in the leaf longevity at 20° C resulted from the decreases in the durations of all of the developmental, mature, and senescent phases. The bulbs without chilling treatment did not sprout leaves and those with chilling treatment sprouted leaves. The increase in the length of chilling treatment from 60 to 120 days affected leaf phenology in same manner as the increase in the growth temperature from 10 to 20° C.     

Paraheliotropic leaf movement in Siratro as a protective mechanism against drought-induced damage to primary photosynthetic reactions: damage by excessive light and heat

Damage to primary photosynthetic reactions by drought, excess light and heat in leaves of Macroptilium atropurpureum Dc. cv. Siratro was assessed by measurements of chlorophyll fluorescence emission kinetics at 77 K (-196°C). Paraheliotropic leaf movement protected waterstressed Siratro leaves from damage by excess light (photoinhibition), by heat, and by the interactive effects of excess light and high leaf temperatures. When the leaves were restrained to a horizontal position, photoinhibition occurred and the degree of photoinhibitory damage increased with the time of exposure to high levels of solar radiation. Severe inhibition was followed by leaf death, but leaves gradually recovered from moderate damage. This drought-induced photoinhibitory damage seemed more closely related to low leaf water potential than to low leaf conductance. Exposure to leaf temperatures above 42°C caused damage to the photosynthetic system even in the dark and leaves died at 48°C. Between 42 and 48°C the degree of heat damage increased with the time of exposure, but recovery from moderate heat damage occurred over several days. The threshold temperature for direct heat damage increased with the growth temperature regime, but was unaffected by water-stress history or by current leaf water status. No direct heat damage occurred below 42°C, but in water-stressed plants photoinhibition increased with increasing leaf temperature in the range 31–42°C and with increasing photon flux density up to full sunglight values. Thus, water stress evidently predisposes the photosynthetic system to photoinhibition and high leaf temperature exacerbates this photoinhibitory damage. It seems probable that, under the climatic conditions where Siratro occurs in nature, but in the absence of paraheliotropic leaf movement, photoinhibitory damage would occur more frequently during drought than would direct heat damage.Abbreviations and symbols PFD photon flux area density - PSI, PSII photosyntem I, II - F _M, F _O, F _V maximum, instantaneous, variable fluorescence emission - PLM paraheliotropic leaf movement; all data of parameter of variation are mean ± standard error     

Growth of Escherichia coli O157:H7 in bruised apple (Malus domestica) tissue as influenced by cultivar, date of harvest, and source

Four of five apple cultivars (Golden Delicious, Red Delicious, McIntosh, Macoun, and Melrose) inoculated with Escherichia coli O157:H7 promoted growth of the bacterium in bruised tissue independent of the date of harvest (i.e., degree of apple ripening) or the source of the apple (i.e., tree-picked or dropped fruit). Apple harvest for this study began 4 September 1998 and ended 9 October, with weekly sampling. Throughout this study, freshly picked (<2 days after harvest) McIntosh apples usually prevented the growth of E. coli O157:H7 for 2 days. Growth of E. coli O157:H7 did occur following 6 days of incubation in bruised McIntosh apple tissue. However, the maximum total cell number was approximately 80-fold less than the maximum total cell number recovered from Red Delicious apples. When fruit was stored for 1 month at 4 degrees C prior to inoculation with E. coli O157:H7, all five cultivars supported growth of the bacterium. For each apple cultivar, the pH of bruised tissue was significantly higher and degrees Brix was significantly lower than the pH and degrees Brix of undamaged tissue regardless of the source. In freshly picked apples, changes in the pH did not occur over the harvest season. Bruised Golden Delicious, McIntosh, and Melrose apple tissue pHs were not significantly different (tree-picked or dropped), and the degrees Brix values of McIntosh, Macoun, and Melrose apple tissue were not significantly different. Single-cultivar preparations of cider did not support growth of E. coli, and the cell concentration of inoculated cider declined over an 11-day test period. The rate of decline in E. coli cell concentration in the McIntosh cider was greater than those in the other ciders tested. The findings of this study suggested that the presence of some factor besides, or in addition to, pH inhibited E. coli growth in McIntosh apples.     

Long-term chilling of young tomato plants under low light and subsequent recovery

The influence of unfavourable climatic conditions at the onset of the growth period on chilling-sensitive tomato (Lycopersicon esculentum Mill., cv. Abunda) was studied by exposing young plants to combinations of low temperature and low light (60–100 mol quanta · m^–2 · s^–1) for several weeks. When the temperature did not decrease below a critical point (8 ° C) no loss of developmental capacity of the plants was detected. However, while new leaves were readily formed upon return to normal growth conditions (22/18 °C, day/night, in a greenhouse), net accumulation of biomass showed a lag phase of approximately one week. This delay was accompanied by a strong, irreversible inhibition of photosynthesis in the fully expanded leaves which had been exposed to the chilling treatment. When plants were subjected to temperatures below 8 ° C, survival rates decreased after three weeks at 6 ° C and irreversible damage of apical meristematic tissue occurred. Drought-hardening prior to chilling ensured survival at 6 ° C and protected the plants against meristem loss.Abreviation Chl chlorophyll Thanks are due to G.P. Telkamp for technical assistance. This research is financially supported by the Netherlands Technology Foundation (STW, Utrecht, The Netherlands), and is coordinated by the Foundation for Biological Research (BION, 's-Gravenhage, The Netherlands).     

The effect of air temperature in successive periods of the growing season on sylleptic shoot formation in young apple trees

In two day-light phytotron rooms young apple trees were exposed to either 15 or 25°C in three successive periods of six weeks (Periods I, II and III) starting in the beginning of the growing season. A control group was kept outside throughout. Exposure to 15°C in either period had little effect on sylleptic growth; there was also hardly any difference with the outside treatment. At 25°C total sylleptic growth was greatly favoured especially when that temperature was applied in all three periods. However, 25°C given continuously did not affect sylleptic shoot number in Period II and III, but applied only in Period II or III greatly enhanced it. In a few cases there was a positive carry-over effect of 25°C given in Period I or II on sylleptic growth in the next period. No such carry-over effect was found for shoot number. Although temperature did influence growth of the parent shoot, that influence was much smaller than for sylleptic growth. The outgrowth of sylleptic shoots mainly occurred in periods when growth vigour of the parent shoot was highest. The distribution of sylleptic shoots along the parent shoot was greatly affected by the period in which the temperature of 25°C was applied suggesting that buds are only able to respond to outgrowth-inducing factors in a certain stage of development.     

Effects of Chilling Temperature on the Activity of Enzymes of Sucrose Synthesis and the Accumulation of Saccharides in Leaves of Three Sugarcane Cultivars Differing in Cold Sensitivity

The effects of short-term exposure to chilling temperature (10 °C) on sucrose synthesis in leaves of the cold-tolerant sugarcane cultivars Saccharum sinense R. cv. Yomitanzan and Saccharum sp. cv. NiF4, and the cold-sensitive cultivar S. officinarum L. cv. Badila were studied. Plants were grown at day/night temperatures of 30/25 °C, and then shifted to a constant day/night temperature of 10 °C. After 52-h exposure to the chilling temperature, sucrose content in the leaves of NiF4 and Yomitanzan showed a 2.5- to 3.5-fold increase relative to that of the control plants that had been left on day/night temperatures of 30/25 °C. No such increase was observed in Badila leaves. Similarly, starch content in the leaves of NiF4 and Yomitanzan was maintained high, but starch was depleted in Badila leaves after the 52-h exposure. During the chilling temperature, sucrose phosphate synthase (SPS; E.C.2.4.1.14) activity was relatively stable in the leaves of NiF4 and Yomitanzan, whereas in Badila leaves SPS activity significantly decreased. There was no significant change in cytosolic fructose-1,6-bisphosphatase activity for the three cultivars at the chilling temperature. This supports the hypothesis that: (1) on exposure to chilling temperature, sucrose content in sugarcane leaves is determined by the photosynthetic rate in the leaves, and is not related to SPS activity; (2) SPS activity in sugarcane leaves at chilling temperature is to be determined by sugar concentration in the leaves.     

Influence of internal sugar levels on apoplasmic retrieval of exogenous sucrose in source leaf tissue

Sugar levels in Beta vulgaris leaves were increased by heat-girdling the petiole and returning the plant to the controlled-environment chamber for 10 and 34 hours. After 10 hours, sucrose influx into the treated leaves was similar to the controls, although sucrose levels increased from 2.1 to 5.3 micromoles per milligram chlorophyll. However, after a 34-hour treatment, sucrose levels increased from 2.1 to 11.5 micromoles per milligram chlorophyll. In this instance, sucrose influx decreased relative to the untreated controls. Decreasing sugar levels by DCMU treatment resulted in a small stimulation of sucrose influx. A similar DCMU treatment applied to leaves of Allium cepa also resulted in an increase in sucrose influx. However, in A. cepa we could not attribute this increase to a lowering of sugar levels, as the kinetic profiles obtained from control leaves did not vary from each other throughout the day, despite considerable changes in sugar levels. Additionally, it appeared that sucrose uptake in onion may be set at some point and remains invariant throughout the day. Similar studies were also conducted on discs cut from mature leaves of Spinacia oleracea var America. Between 1 and 8 hours after the onset of the photoperiod, the sucrose content of the spinach leaves increased from 2.6 to 9.3 micromoles per milligram chlorophyll. A comparison of the kinetic profiles obtained from leaf discs, taken at these times, indicated that sucrose uptake was not influenced by these changes in internal sugar levels. The relationship between the above findings and `trans' inhibition of exogenous sucrose uptake is discussed. Although intermediate changes in sugar levels in sugar beet leaves did not appear to affect sucrose influx, autoradiographic studies revealed that these changes dramatically affected the partitioning of exogenously supplied [¹⁴C]sucrose. Our results indicate that while intermediate changes in internal sugar levels have little effect on sucrose influx across the plasmalemma, they may dramatically affect partitioning between the phloem and the mesophyll vacuole.     

Spring behavioral patterns of the apple blossom weevil

Individual pairs of overwintered adult apple blossom weevils, Anthonomus pomorum (L.), confined with apple twigs under different ambient temperatures in the laboratory and on apple trees in the field, were observed through day and night for their spring activities. Flight behavior in relation to ambient temperature was also investigated under laboratory conditions using flight stands. Both sexes displayed predominantly nocturnal behavior patterns in both the laboratory and the field. Feeding, crawling, and mating activities increased following sunset in the field or onset of scotophase in the laboratory while resting occurred most frequently during daylight hours. Results of the laboratory experiments showed that temperature affected significantly the activity patterns. The diel pattern of activities became less distinctive at higher temperatures (above 15°C), and total activities in crawling, feeding, and mating were suppressed significantly at lower temperatures (below 5°C). Over 97% of the test weevils initiated take-off response from flight stands at 20°C within the 30 min trial period; however, flight initiation rarely occurred at temperatures 12°C or below. Overall, results of the laboratory and field experiments indicate that A. pomorum is a remarkably cold-adapted insect with ability to crawl, feed, and mate at a few degrees above freezing, a physiological attribute necessary for the exploitation of early stages of apple bud development in the cold early spring.     

Up-regulation of sucrose phosphate synthase in rice grown under elevated CO2 and temperature

Rice (Oryza sativa L. cv. IR-30) was grown season-long in outdoor, controlled-environment chambers at 33 Pa CO₂ with day/night/paddy-water temperatures of 28/21/25 °C, and at 66 Pa CO₂ with five different day/night/paddy-water temperature regimes (25/18/21, 28/21/25, 31/24/28, 34/27/31 and 37/30/34 °C). Sucrose phosphate synthase (SPS) activities in leaf extracts at 21, 48 and 81 days after planting (DAP) were assayed under saturating and selective (limiting) conditions. Diel SPS activity data indicated that rice SPS was light regulated; with up to 2.2-fold higher rates during the day. Throughout the growth season, leaf SPS activities were up-regulated in the CO₂-enriched plants, averaging 20 and 12% higher than in ambient-CO₂ grown plants in selective and saturating assays, respectively. Similarly, SPS activities increased 2.4% for each 1 °C rise in growth temperature from 25 to 34 °C, but de creased 11.5% at 37 °C. Leaf sucrose content was higher, and mirrored SPS activity better, than starch, although starch was more responsive to CO₂ treatment. Leaf sucrose and starch contents were significantly higher throughout the season in plants at elevated CO₂, but the N content averaged 6.5% lower. Increasing growth temperatures from 25 to 37 °C caused a linear decrease (62%) in leaf starch content, but not in sucrose. Consequently, the starch:sucrose ratio declined with growth temperature. The data are consistent with the hypothesis that the up-regulation of leaf SPS may be an acclimation response of rice to optimize the utilization and export of organic-C with the increased rates of inorganic-C fixation in elevated CO₂ or temperature growth regimes.     

Influence of root temperature on the potassium requirements of young tomato plants

The effect of low root temperature on the growth and K requirements of young tomato (Lycopersicon esculentum Mill. cv Sonatine) plants was investigated. When K was supplied in solution at high concentration (5 mM), lowering the temperature of the root system from 25° to 15°C reduced the relative growth rate so that after ten days plant dry weight was 60% and leaf area 44% of that of controls maintained at 25°C. Shoot: root dry weight ratio was initially increased by cooling, but declined to 84% of controls after ten days. In spite of these changes in shoot: root ratio the concentration of K in whole plants, expressed on the basis of tissue water, was stable throughout the experiment and was significantly higher than that of controls. Further, the critical concentration of K for shoots (the concentration in the shoot associated with 90% maximum growth) was also increased at root temperatures of 15° and 30°C compared with 24°C. It is suggested that the higher concentration of K at low root temperature may reflect a real increase in requirement for the element at the physiological level. Preliminary measurements of the solute potential demonstrated a less osmotically active sap in leaves of root-cooled plants, thus there may be a greater reliance on K as an osmoticum in these individuals. When supplies of K limited growth, root-cooling had no effect on any of the parameters determining the efficiency of its use; the minimum concentration to which roots could deplete the solution of K was identical for cooled and control plants and at the same stage of visible deficiency there was no significant difference in the efficiency ratio (mg DW, mg^-1 K) or utilization efficiency (mg DW mM ^-1 K), in spite of large differences in the partitioning of dry matter.     

Apple leaf senescence: leaf disc compared to attached leaf

Attached apple leaves (Pyrus malus L., Golden Delicious) began to lose protein in early August as the first sign of senescence. Apple leaf discs prepared from samples before early August gained protein for up to 7 days after detachment. After early August, the loss of protein from leaf discs was no greater than the loss from attached leaves in 7 days. The loss of chlorophyll from leaf discs began over 2 months before attached leaves began to lose chlorophyll naturally and before leaf discs lost protein. Leaf discs from presenescent leaves did not senesce significantly faster when maintained in darkness instead of 12 hours of light. In general, the loss of protein and chlorophyll from apple leaf discs after 7 days was much less than for most other leaf types studied.