Two different strategies of Mediterranean macchia plants to avoid photoinhibitory damage by excessive radiation levels during summer drought

The adaptive strategies to high radiation and water stress of the drought tolerant evergreen sclerophylls Quercus coccifera and Arbutus unedo are compared to those of the semi-deciduous Cistus spp. (C. albidus and C. monspeliensis). Cistus spp. partially avoided drought by a marked reduction of their transpirational surface through leaf abscission during summer, when predawn water potential declined below -5.5 MPa. Chlorophyll fluorescence measurements revealed a reversible diurnal decrease of maximum photochemical efficiency of PSII (F_v/F_m), which became more accentuated during summer drought in all species. An important strategy to avoid damage by excessive radiation levels in Cistus spp. was the structural regulation of light interception through leaf angle changes, from a more horizontal orientation in spring (< 35°) to a more vertical orientation in summer (> 70°). Horizontal orientated leaves were highly susceptible to photoinhibition, and excessive radiation often resulted in irreversible photodamage followed by leaf abscission during summer, whereas vertical leaf orientation appeared to protect the leaf from severe photoinhibition. Still, these mechanisms were not fully successful in avoiding chronic photoinhibition, and predawn F_v/F_m values remained low in Cistus spp. during summer (only exhibiting a partial overnight recovery). Evergreen sclerophylls were less susceptible to photoinhibition, and the diurnal decline in F_v/F_m remained fully reversible during drought. Structural regulation of light interception was not found to be an important strategy in these species, and only small, though significant changes in leaf angle occurred. The ecological importance of the adaptive strategies of each functional group is discussed.     

Abscission: the initial effect of ethylene is in the leaf blade

The leaf blade of cotton (Gossypium hirsutum L. cv. Stoneville 213) was investigated as the initial site of ethylene action in abscission. Ethylene applied at 14 μl/l to intact 3-week-old plants caused abscission of the third true leaf within 3 days. However, keeping only the leaf blade of this leaf in air during ethylene treatment of the rest of the plant completely prevented its abscission for up to 7 days. This inhibition of abscission was apparently the result of continued auxin production in the blade since (a) the application of an auxin transport inhibitor to the petiole of the air-treated leaf blade restored ethylene sensitivity to the leaf in terms of abscission; (b) repeated applications of naphthaleneacetic acid to the leaf blade of the third true leaf, when the entire plant was exposed to ethylene, had the same preventive effect on abscission of this leaf as keeping its leaf blade in air; and (c) the inhibitory effect of ethylene on auxin transport in the petiole, which is reduced by auxin treatment, was also reduced by placing the leaf blade in air.     

Involvement of ethylene in the action of the cotton defoliant thidiazuron

The effect of the defoliant thidiazuron (N-phenyl-N′-1,2,3-thiadiazol-5-ylurea) on endogenous ethylene evolution and the role of endogenous ethylene in thidiazuron-mediated leaf abscission were examined in cotton (Gossypium hirsutum L. cv Stoneville 519) seedlings. Treatment of 20- to 30-day-old seedlings with thidiazuron at concentrations equal to or greater than 10 micromolar resulted in leaf abscission. At a treatment concentration of 100 micromolar, nearly total abscission of the youngest leaves was observed. Following treatment, abscission of the younger leaves commenced within 48 hours and was complete by 120 hours. A large increase in ethylene evolution from leaf blades and abscission zone explants was readily detectable within 24 hours of treatment and persisted until leaf fall. Ethylene evolution from treated leaf blades was greatest 1 day posttreatment and reached levels in excess of 600 nanoliters per gram fresh weight per hour (26.7 nanomoles per gram fresh weight per hour). The increase in ethylene evolution occurred in the absence of increased ethane evolution, altered leaf water potential, or decreased chlorophyll levels. Treatment of seedlings with inhibitors of ethylene action (silver thiosulfate, hypobaric pressure) or ethylene synthesis (aminoethoxyvinylglycine) resulted in an inhibition of thidiazuron-induced defoliation. Application of exogenous ethylene or 1-aminocyclopropane-1-carboxylic acid largely restored the thidiazuron response. The results indicate that thidiazuron-induced leaf abscission is mediated, at least in part, by an increase in endogenous ethylene evolution. However, alterations of other phytohormone systems thought to be involved in regulating leaf abscission are not excluded by these studies.     

Anatomy of leaf abscission in the Amur honeysuckle (Lonicera maackii, Caprifoliaceae): a scanning electron microscopy study

Lonicera maackii (Rupr.) Maxim. (Amur honeysuckle) is native to Asia and an important ornamental in China. However, the anatomy of leaf abscission (shedding) in L. maackii had not been studied previously. Such work is needed not only because knowledge of the leaf abscission process is important for a horticultural species like L. maackii but also because leaf abscission is probably the least understood abscission process, as it occurs so rapidly. Therefore, our objective was to use scanning electron microscopy (SEM) to examine the progression of leaf abscission in L. maackii at the cellular level. L. maackii branches with leaves were regularly collected in Beijing, China over the 2-month period in which leaves abscise, and examined with SEM. We found that, unlike in model species, the cortex is involved in abscission, forming an “abaxial gap.” We discovered that there is no discrete abscission zone prior to the onset of abscission and that no cell divisions precede abscission. An abscission zone did become evident well after the abscission process had begun, but its cells were enlarged, not constricted as in typical abscission zones. In the abaxial gap, intact cells separated at their middle lamella, but in the abscission zone, cell separation involved the entire wall, which is not typical. We did observe expected mechanical fission of vascular tissues. While the leaf abscission process we observed in L. maackii has similarities with model systems, aspects deviate from the expected.     

Effects of morphactin and other auxin transport inhibitors on soybean senescence and pod development

Because triiodobenzoic acid increases pod number, albeit variably, in soybean (Glycine max), we tested other auxin-transport inhibitors. Morphactins, especially methylchlorflurenol (MCF), were found to be very active (optimal concentration 10 micromolar) when sprayed onto the foliage. Applications at 1 week after the start of flowering were most effective, producing a 40% increase in pod number with little inhibition (12%) of stem elongation. MCF increased the number of pods initiated (reaching 1 cm length) at least partially by prolonging the initiation period, while pod abortion (failure of pods > 1 cm long) remained low. Generally, MCF did not increase seed yield (dry weight/plant); more, but smaller seeds, were formed by the treated plants. The promotive effect of MCF on pod initiation seems to be independent of its inhibition of stem elongation, which is insignificant at 10 micromolar. MCF delayed pod maturation by 3 to 4 days, while foliar yellowing, blade abscission, and petiole abscission were retarded by 2, 4, and 2 days, respectively. MCF has only a small effect on senescence and that could be indirect, due to a delay in pod development. Other auxin-transport inhibitors tested, including N-1-naphthylphthalamic acid, produced little or no increase in pod number; however, 0.1 millimolar 5-[2′-carboxyphenyl]-3-phenylpyrazole caused a 27% increase. These results implicate auxin as a potential regulator of pod development, and they show that soybean seed yield is not simply sink limited.     

Auxin Transport as Related to Leaf Abscission during Water Stress in Cotton

Plant water deficits reduced the basipetal transport of auxin in cotyledonary petiole sections taken from cotton (Gossypium hirsutum L.) seedings. A pulse-labeling technique was employed to eliminate complications of uptake or exit of ¹⁴C-indoleacetic acid from the tissue. The transport capacity or the relative amount of radioactivity in a 30-minute pulse which was basipetally translocated was approximately 30% per hour in petioles excised from well watered seedlings (plant water potentials of approximately -4 to -8 bars). No cotyledonary leaf abscission took place in well watered seedlings. Plant water potentials from -8 to -12 bars reduced the transport capacity from 30 to 15% per hour, and although the leaves were wilted, cotyledonary abscission did not increase appreciably at these levels of stress. The threshold water potential sufficient to induce leaf abscission was approximately -13 bars and abscission increased with increasing stress while the auxin transport capacity of the petioles remained relatively constant (15% per hour). The basipetal transport capacity of well watered petioles tested under anaerobic conditions and acropetal transport tested under all conditions were typically less than basipetal transport under the most severe stress conditions. Cotyledonary abscission took place during and 24 hours after relief of stress with little or no abscission taking place 48 hours after relief of stress. Although the water potential returned to -4 bars within hours after rewatering the stressed plants, partial recovery of the basipetal transport capacity of the petioles was not apparent until 48 hours after rewatering, and at least 72 hours was required to return the transport capacity to near normal values. These data support the view that decreased levels of auxin reaching the abscission zone from the leaf blade influence the abscission process and further suggest that the length of time that the auxin supply is maximally reduced is more critical than the degree of reduction.     

Ethylene Evolution following Treatment with 1-Aminocyclopropane-1-carboxylic Acid and Ethephon in an in Vitro Olive Shoot System in Relation to Leaf Abscission

1-Aminocyclopropane-1-carboxylic acid (ACC) supplied via the cut base of detached olive shoots caused a burst of ethylene from leaves, but other cyclopropanes tested did not exhibit this effect. Ethephon (ET) and another ethylene-releasing compound caused a prolonged increase in ethylene evolution. ACC had only a very limited effect on leaf abscission regardless of concentration, whereas shoots placed with cut bases in ET for 60 to 80 minutes exhibited 100% leaf abscission within 90 hours. Shoots with inflorescences treated with ET just prior to anthesis began to wilt in vitro within 20 to 30 hours and failed to exhibit leaf abscission. At earlier stages of development, ET induced more leaf abscission on reproductive shoots than on vegetative shoots. It is suggested that the duration of ethylene evolution from the leaves governs their potential for abscission and that bursts of ethylene evolution even though large in amount may not induce abscission.     

Starch Synthetase, Phosphorylase, ADPglucose Pyrophosphorylase, and UDPglucose Pyrophosphorylase in Developing Maize Kernels

Three types of whole plant experiments are presented to substantiate the concept that an important function of ethylene in abscission is to reduce the transport of auxin from the leaf to the abscission zone. (a) The inhibitory effect of ethylene on auxin transport, like ethylene-stimulated abscission, persists only as long as the gas is continuously present. Cotton (Gossypium hirsutum L. cv. Stoneville 213) and bean (Phaseolus vulgaris L. cv. Resistant Black Valentine) plants placed in 14 μl/l of ethylene for 24 or 48 hours showed an increase in leaf abscission and a reduced capacity to transport auxin; but when returned to air, auxin transport gradually increased and abscission ceased. (b) Ethylene-induced abscission and auxin transport inhibition show similar sensitivities to temperature. A 24-hour exposure of cotton plants to 14 μl/l of ethylene at 8 C resulted in no abscission and no significant inhibition of auxin transport. Increasing the temperature during ethylene treatment resulted in a progressively greater reduction in auxin transport with abscission occurring at [unk]27 C where auxin transport was inhibited over 70%. (c) Auxin pretreatment reduced both ethylene-induced abscission and auxin transport inhibition. No abscission occurred, and auxin transport was inhibited only 18% in cotton plants which were pretreated with 250 mg/l of naphthalene acetic acid and then placed in 14 μl/l of ethylene for 24 hours. In contrast, over 30% abscission occurred, and auxin transport was inhibited 58% in the corresponding control plants.     

Synthesis of Cellulase during Abscission of Phaseolus vulgaris Leaf Explants

When abscission in leaf explants from Phaseolus vulgaris, cultivar Red Kidney, was allowed to proceed while the explants were in ²H₂O, a 1.25% increase in the buoyant density of cellulase in a cesium chloride gradient was observed. These data indicate that the increase in cellulase activity during abscission is a result of the synthesis of new protein. Two differentially soluble forms of cellulase are present in the abscission zone. The form which is soluble only in a high salt buffer seems more closely related to the abscission process than the form which is soluble in dilute buffer. The correlation between changes in pull force and increase in cellulase activity and the effects of several hormones on cellulase activity are discussed.     

Induction of resistance to dark abscission by malformin in white light

When cuttings or seedlings of Phaseolus aureus were treated proximally with malformin for 2 days in continuous white light, resistance to subsequent leaf abscission in the dark resulted. The amount of resistance diminished as the concentration of malformin decreased from 10 to 0.1 micromolar. Resistance to dark abscission persisted for 7 days in continuous light. Little resistance was obtained when cuttings were taken from seedlings grown under low irradiance and short photoperiods, but resistance gradually increased as the photoperiod increased. Resistance to dark abscission induced by malformin in light differs from inhibition of abscission by indoleacetic acid because when malformin is applied in the dark it stimulates abscission after distal or proximal application. Malformin induces resistance only in conjunction with light treatment.     

Transdifferentiation of Mature Cortical Cells to Functional Abscission Cells in Bean

Abscission explants of bean (Phaseolus vulgaris L.) were treated with ethylene to induce cell separation at the primary abscission zone. After several days of further incubation of the remaining petiole in endogenously produced ethylene, the distal two-thirds of the petiole became senescent, and the remaining (proximal) portion stayed green. Cell-to-cell separation (secondary abscission) takes place precisely at the interface between the senescing yellow and the enlarging green cells. The expression of the abscission-associated isoform of β-1,4-glucanhydrolase, the activation of the Golgi apparatus, and enhanced vesicle formation occurred only in the enlarging cortical cells on the green side. These changes were indistinguishable from those that occur in normal abscission cells and confirm the conversion of the cortical cells to abscission-type cells. Secondary abscission cells were also induced by applying auxin to the exposed primary abscission surface after the pulvinus was shed, provided ethylene was added. Then, the orientation of development of green and yellow tissue was reversed; the distal tissue remained green and the proximal tissue yellowed. Nevertheless, separation still occurred at the junction between green and yellow cells and, again, it was one to two cell layers of the green side that enlarged and separated from their senescing neighbors. Evaluation of Feulgen-stained tissue establishes that, although nuclear changes occur, the conversion of the cortical cell to an abscission zone cell is a true transdifferentiation event, occurring in the absence of cell division.     

The Anticyclic Timing of Leaf Senescence in the Parasitic Plant Viscum album Is Closely Correlated with the Selective Degradation of Sulfur-Rich Viscotoxins

Leaf senescence and abscission have been studied in the semi-parasitic plant mistletoe (Viscum album). Leaf senescence and abscission occur in the summer, when the metabolic activity of the host has reached its maximum. In contrast with their hosts, mistletoes selectively degrade only one major leaf protein during leaf senescence, the sulfur-rich viscotoxin, whereas most of the remaining leaf proteins are lost during abscission. The changes in viscotoxin content are paralleled by changes in the concentration of the corresponding mRNA. Shortly before the onset of leaf senescence, the mRNA for viscotoxin has disappeared from the leaves. The anticyclic timing of leaf senescence and the degradation of only one major leaf protein seems to reflect an adaptation of the parasite to its habitat.     

Ethylene-induced leaf abscission in cotton seedlings : the physiological bases for age-dependent differences in sensitivity

The speed of ethylene-induced leaf abscission in cotton (Gossypium hirsutum L. cv LG-102) seedlings is dependent on leaf position (i.e. physiological age). Fumigation of intact seedlings for 18 hours with 10 microliters per liter of ethylene resulted in 40% abscission of the still-expanding third true (3°) leaves but had no effect on the fully expanded first true (1°) leaves. After 42 hours of fumigation with 50 microliters per liter of ethylene, total abscission of the 3° leaves occurred while <50% abscission of the 1° leaves was observed. On a leaf basis, endogenous levels of free IAA in 1° leaves were approximately twice those of 3° leaves. Free IAA levels were reduced equally (approximately 55%) in both leaf types after 18 hours of ethylene (10 microliters per liter) treatment. Ethylene treatment of intact seedlings inhibited the basipetal movement of [¹⁴C]IAA in petiole segments isolated from both leaf types in a dose-dependent manner. The auxin transport inhibitor N-1-naphthylphthalamic acid increased the rate and extent of ethylene-induced leaf abscission at both leaf positions but did not alter the relative pattern of abscission. Abscission-zone explants prepared from 3° leaves abscised faster than 1° leaf explants when exposed to ethylene. Ethyleneinduced abscission of 3° explants was not appreciably inhibited by exogenous IAA while 1° explants exhibited a pronounced and protracted inhibition. The synthetic auxins 2,4-D and 1-naphthaleneacetic acid completely inhibited ethylene-induced abscission of both 1° and 3° explants for 40 hours. It is proposed that the differential abscission response of cotton seedling leaves is primarily a result of the limited abscission-inhibiting effects of IAA in the abscission zone of the younger leaves.     

The never ripe mutation blocks ethylene perception in tomato.

Seedlings of tomato fruit ripening mutants were screened for their ability to respond to ethylene. Ethylene induced the triple response in etiolated hypocotyls of all tomato ripening mutants tested except for one, Never ripe (Nr). Our results indicated that the lack of ripening in this mutant is caused by ethylene insensitivity. Segregation analysis indicated that Nr-associated ethylene insensitivity is a single codominant trait and is pleiotropic, blocking senescence and abscission of flowers and the epinastic response of petioles. In normal tomato flowers, petal abscission and senescence occur 4 to 5 days after the flower opens and precede fruit expansion. If fertilization does not occur, pedicel abscission occurs 5 to 8 days after petal senescence. If unfertilized, Nr flowers remained attached to the plant indefinitely, and petals remained viable and turgid more than four times longer than their normal counterparts. Fruit development in Nr plants was not preceded by petal senescence; petals and anthers remained attached until they were physically displaced by the expanding ovary. Analysis of engineered 1-aminocyclopropane-1-carboxylate (ACC) synthase-overexpressing plants indicated that they are phenotypic opposites of Nr plants. Constitutive expression of ACC synthase in tomato plants resulted in high rates of ethylene production by many tissues of the plant and induced petiole epinasty and premature senescence and abscission of flowers, usually before anthesis. There were no obvious effects on senescence in leaves of ACC synthase overexpressers, suggesting that although ethylene may be important, it is not sufficient to cause tomato leaf senescence; other signals are clearly involved.     

Cassava Response to Water Deficit in Deep Pots: Root and Shoot Growth, ABA, and Carbohydrate Reserves in Stems, Leaves and Storage Roots

Cassava (Manihot esculenta, Crantz) is an important staple crop for tropical climates worldwide, including drought-prone environments where it is valued for its reliable yield. The extent to which stress tolerance involves regulation of growth and carbon balance aided by remobilization of carbohydrate from various plant parts was investigated. Plants were grown in 1-meter high pots to permit observation of deep rooting while they were subjected to four soil water regimes over a 30-d period. Transpiration declined abruptly in conjunction with leaf ABA accumulation and severe leaf abscission. In water stressed plants, growth of all plant parts decreased substantially; however, a basal rate of leaf growth continued to provide some new leaves, and although growth of fibrous lateral roots was reduced, main root elongation to deeper regions was only modestly decreased by stress. In leaf blades and petioles, sugars were the predominant form of nonstructural carbohydrate and about one third was in starch; these reserves were depleted rapidly during stress. In contrast, stems and storage roots maintained relatively high starch concentrations and contents per organ until final harvest. Stems gradually lost starch and had sufficient reserves to serve as a prolonged source of remobilized carbohydrate during stress. The amount of starch stored in stems represented about 35 % of the reserve carbohydrate in the plant at the onset of water stress (T₀), and 6 % of total plant dry mass. We suggest that this pool of carbohydrate reserves is important in sustaining meristems, growing organs, and respiring organs during a prolonged stress and providing reserves for regrowth upon resumed rainfall.     

Citrus leaf abscission. Regulatory role of exogenous auxin and ethylene on peroxidases and endogenous growth substances

Abstract The abscission of citrus leaf explants demonstrates the well-known enhancing effect of ethylene and the delaying one of auxin when treatment is started at excision time. Total peroxidase activity increases differently in tissues of the blade, abscission zone, and petiole. The highest activity at zero time is recovered in abscission zone in which also the response to the abscission regulators is the most visible. Isoperoxidase profiles are modified in opposite directions by ethylene and auxin respectively. Both regulators affect the activity of the same cathodic and anodic isoperoxidases without any qualitative changes. By the same time, auxin-like compounds increase in isolated abscission zones at 24 h from excision and decrease at 48 h. The level of one inhibitor complex undergoes an inverse variation. It is suggested that the increase in auxin during the first stage of abscission is necessary for influencing the growth of cells which is required to cause abscission.     

Drought stress and recovery of riparian cottonwoods due to water table alteration along Willow Creek,Alberta

A 5-m-deep gravel pit was excavated from 1996 to 1998 in the floodplain between Willow Creek, Alberta, and a grove of balsam poplars ('cottonwoods', Populus balsamifera L.) and water level at the pit was lowered 2.5 m through pumping. This interrupted the infiltration of stream water into the riparian groundwater and imposed drought stress on the cottonwoods. Trees in the drought-affected grove displayed extensive leaf senescence and abscission in late August 1998, while trees in nearby control groves remained green until autumnal senescence in late September. The precocious senescence was accompanied by a two-thirds reduction in leaf stomatal conductance (g _s) but mid-day leaf xylem water potentials (ψ_l) were only slightly reduced (?1.55 vs 1.42 MPa). Pumping ceased in 1999, the pit was partially refilled, and the hydraulic linkage between the stream and the riparian zone recovered. Subsequently in August 1999, g _s and ψ_l were similar for trees in the affected and control groves and senescence phenologies were similar in 1999 and 2000. Annual branch growth increments varied 3-fold across years between 1994 and 1999, but there was no reduction in these growth increments in the drought-affected trees in 1998 or 1999. This study supports the hydraulic linkage between a stream and the adjacent riparian zone in a semi-arid region and demonstrates the vulnerability of riparian cottonwoods to drought due to water table depletion. It also indicates rapid physiological recovery of cottonwoods following restoration of water availability.