Induced sink strength as a prerequisite for induced tannin biosynthesis in developing leaves of Populus

Induced defenses occur predominately in young, developing plant tissues that rely upon carbohydrate import to support their growth and development. To test the hypothesis that the induced production of carbon-based defenses is dependent upon photoassimilate import, we examined the response of developing leaves of hybrid poplar (Populus deltoides 2 P. nigra) saplings to wounding by gypsy moth caterpillars (Lymantria dispar L.) and exogenous jasmonic acid (JA). Growth rates, condensed tannin contents and acid invertase activities were measured for individual leaves and the translocation of ¹³C-labeled resources between orthostichous source-sink pairs was quantified. Results showed a substantial increase in the activity of cell wall invertase in sink leaves wounded by gypsy moth caterpillars and treated with JA. JA-induced sink leaves also imported 3-4 times as much ¹³C-labeled carbon from orthostichous source leaves relative to controls and allocated a significant portion of this imported ¹³C to condensed tannin biosynthesis. Reduced carbohydrate flow to these leaves, caused by source leaf removal, resulted in reduced condensed tannin levels and the emergence of a growth-defense tradeoff. These results indicate that (1) induced sink strength is elicited by insect wounding and JA application in hybrid poplar foliage, (2) imported resources are allocated to the production of carbon-based defenses, and (3) the level of induced defense in leaves can be constrained by the ability of leaves to import carbohydrates from source tissues. Together, these results suggest that within-canopy variations in induced resistance may arise in part because of uneven distribution of resources to induced foliage.     

Is polyphenol induction simply a result of altered carbon and nitrogen accumulation?

Carbon translocation in plants is shaped by phyllotaxis and regulated by source/sink interactions that respond to the demands of growth and defense. We have studied this extensively in poplar saplings, and recently showed that unlike carbon import, nitrogen is not translocated to sink leaves in response to application of jasmonic acid. Here we report that this is also true for young trees in the field. We discuss the importance of transport processes in establishing local C:N ratios, and suggest that the JA-induced flow of C but not N to sink tissues, and their corresponding increases in C-based defenses, may simply reflect a plant adaptation to handle excess reduced carbon and energy.     

A preliminary investigation of the role of auxin and cytokinin in sylleptic branching of three hybrid poplar clones exhibiting contrasting degrees of sylleptic branching

Sylleptic branches grow out from lateral buds during the same growing season in which the buds are formed. This type of branching is present in poplar and in many tropical species. It results in the production of more branches, more leaves and expanded photosynthetic capacity and is thought to assist in increasing the overall growth and biomass of the tree at a young age. However, very little is known about the physiology of sylleptic branching in poplar, which is an extremely important source of fibre and fuel. In the present study of three hybrid poplar clones (11-11, 47-174 and 49-177) of Populus trichocarpa x P. deltoides exhibiting contrasting degrees of sylleptic branching, an analysis was carried out on parent shoot elongation and sylleptic branching, together with a preliminary comparison of the parent shoots' sensitivity to auxin (naphthaleneacetic acid) as a repressor of lateral bud outgrowth, and cytokinin (benzyladenine) as a promoter. Suggestive evidence was found for an inverse correlation between parent shoot sensitivity to auxin and the degree of sylleptic branching, as well as a partially positive correlation with respect to sensitivity to cytokinin. The present data are consistent with the hypothesis that auxin and cytokinin may play repressive and promotive roles, respectively, in the sylleptic branching of hybrid poplar.     

Growth pattern ofAbies mariesii saplings under conditions of open-growth and suppression

The growth pattern was analysed forAbies mariesii Mast. saplings in the subalpine climax forests of the northern Yatsugatake Mountains, Central Japan. The lateral branches were distinguished from the trunk not only with respect to the branching habits and needle arrangement but also with respect to the distribution of dry matter to needles, the slenderness of shoot-stem and the longevity of needles on them. Based on these differences, the contribution of the trunk and lateral branches to the matter production function of saplings is discussed. The lateral branches showed plastic changes in their growth characters in order to expand and maintain an assimilation system effectively, under conditions of suppression. In addition, the trunk growth was inhibited compared with the lateral branch growth and the apical dominance among lateral branches was weakened in the suppressed saplings. It was concluded that these growth habits resulted in the observed difference in the crown shape of saplings in the different habitats. Arguments are presented regarding the adaptive meanings of such a growth pattern for the shade tolerance and survival of the saplings and for the regeneration of the forest.     

The impact of reduced vacuolar invertase activity on the photosynthetic and carbohydrate metabolism of tomato

The impact of reduced vacuolar invertase activity on photosynthetic and carbohydrate metabolism was examined in tomato (Solanum lycopersicon L.). The introduction of a co-suppression construct (derived from tomato vacuolar invertase cDNA) produced plants containing a range of vacuolar invertase activities. In the leaves of most transgenic plants from line INV-B, vacuolar invertase activity was below the level of detection, whereas leaves from line INV-A and untransformed wild-type plants showed considerable variation. Apoplasmic invertase activity was not affected by the co-suppression construct. It has been suggested that, in leaves, vacuolar invertase activity regulates sucrose content and its availability for export, such that in plants with high vacuolar invertase activity a futile cycle of sucrose synthesis and degradation takes place. In INV-B plants with no detectable leaf vacuolar invertase activity, sucrose accumulated to much higher levels than in wild-type plants, and hexoses were barely detectable. There was a clear threshold relationship between invertase activity and sucrose content, and a linear relationship with hexose content. From these data the following conclusions can be drawn. (i) In INV-B plants sucrose enters the vacuole where it accumulates as hydrolysis cannot take place. (ii) There was not an excess of vacuolar invertase activity in the vacuole; the rate of sucrose hydrolysis depended upon the concentration of the enzyme. (iii) The rate of import of sucrose into the vacuole is also important in determining the rate of sucrose hydrolysis. The starch content of leaves was not significantly different in any of the plants examined. In tomato plants grown at high irradiance there was no impact of vacuolar invertase activity on the rate of photosynthesis or growth. The impact of the cosuppression construct on root vacuolar invertase activity and carbohydrate metabolism was less marked.Abbreviations CaMV Cauliflower Mosaic Virus - WT wild type     

Can saliva from moose,Alces alces,affect growth responses in the sallow,Salix caprea?

Herbivores have the potential to affect the morphological traits of the plants they feed on, such as the breaking of apical dominance that occurs when shoot tips are browsed, leading to increased lateral branching and decreased competition among growing points. Despite such effects, however, it remains unclear whether components in the saliva of the browsing animals are directly involved in amplifying and/or reducing the plant growth responses. To test this possibility, a controlled experiment was performed in which herbivory by moose ( Alces alces ) was simulated by tearing 60 sallow saplings ( Salix caprea ) with a moose jaw. Of these 60, half were also treated with moose saliva. Selected growth traits of the torn plants with and without saliva treatment were examined directly after treatment (week 0) and 1, 2, 4, 6, 8, 10, 12 and 15 weeks after the tearing. Also included in the study was an untreated (no tearing, no saliva) control group of another 30 sallow saplings. Traits monitored on the saplings were height and the number of branches, buds and leaves. Application of moose saliva on torn saplings led to significantly more branches than on saplings without saliva treatment. Moose saliva, however, had no significant effects on the other growth traits, although for most of these characteristics both groups of torn saplings showed increased values compared to the undamaged control. Overall, these results demonstrate that moose saliva has a consistent stimulatory effect on branching of sallow saplings but not on the other growth traits examined in this study.     

Comparative investigations on the distribution of sucrose synthase activity and invertase activity within growing, mature and old leaves of some C3 and C4 plant species

Sucrose synthase (EC 2.4.1.13) activity in young growing leaves was highest in the leaf base in eggplants ( Solanum melongena L.), cassava ( Manihot esculenta Crantz), grapevine ( Vitis vinifera L.), and in the leaf sheath of sugar cane ( Saccharum of ficinarum L.) and maize ( Zea mays L.). In addition, increasing sucrose synthase activity was measured towards the edge of growing eggplant leaves while the activity in mature leaves was highest in the midrib. The activity of acid and alkaline invertase was very low in the midrib but higher in the blade of fully expanded eggplant leaves. Highest invertase activities were found in younger growing leaves. It was concluded that in growing leaves a close relationship might exist between the activity of sucrose synthase and the import of sucrose from source leaves.
Detachment of mature eggplant leaves led to a 2- to 3-fold increase of sucrose synthase activity in blade and midrib of these leaves. In contrast, invertase activity decreased after detachment in both leaf blade and midrib. It was concluded that the rise in sucrose synthase activity might have been caused by the observed increase of sucrose concentration in detached leaves and that sucrose synthase might have an important role in the regulation of sucrose content of the conducting tissue.     

Branch architecture,light interception and crown development in saplings of a plagiotropically branching tropical tree,Polyalthia jenkinsii (Annonaceae)

To investigate crown development patterns, branch architecture, branch-level light interception, and leaf and branch dynamics were studied in saplings of a plagiotropically branching tree species, Polyalthia jenkinsii Hk. f. & Thoms. (Annonaceae) in a Malaysian rain forest. Lengths of branches and parts of the branches lacking leaves ('bare' branches) were smaller in upper branches than in lower branches within crowns, whereas lengths of 'leafy' parts and the number of leaves per branch were larger in intermediate than in upper and lower branches. Maximum diffuse light absorption (DLA) of individual leaves was not related to sapling height or branch position within crowns, whereas minimum DLA was lower in tall saplings. Accordingly, branch-level light interception was higher in intermediate than in upper and lower branches. The leaf production rate was higher and leaf loss rate was smaller in upper than in intermediate and lower branches. Moreover, the branch production rate of new first-order branches was larger in the upper crowns. Thus, leaf and branch dynamics do not correspond to branch-level light interception in the different canopy zones. As a result of architectural constraints, branches at different vertical positions experience predictable light microenvironments in plagiotropic species. Accordingly, this pattern of carbon allocation among branches might be particularly important for growth and crown development in plagiotropic species.     

外源茉莉酸诱导的青杨生化抗性及其对舞毒蛾幼虫食物利用的影响

【目的】为探讨外源茉莉酸(jasmonic acid, JA)诱导的青杨 Populus cathayana Rehd.抗性对舞毒蛾 Lymantria dispar (L.)幼虫食物利用的影响。【方法】在室内对青杨苗木喷施0.001和0.1 mmol/L两种浓度的茉莉酸,对照喷0.2%的丙酮水溶液,喷施后1, 5和10 d分别采集叶片,分析其初生和次生物质含量的变化。另选喷施0.1 mmol/L茉莉酸的青杨苗木,喷施后1, 5和10 d分别接舞毒蛾2龄幼虫,单头饲养,测定其取食量、体重和排粪量及发育历期,统计分析其食物消化率、转化率和利用率。【结果】两种浓度茉莉酸处理均使青杨叶片中的蛋白质和可溶性糖含量降低,而木质素、单宁、黄酮和酚类物质含量增加。蛋白质和可溶性糖在处理第10天时含量最低,其中高浓度茉莉酸处理较对照分别降低46.5%和49.1%,低浓度处理分别降低30.6%和22.8%。叶片中酚类物质含量在高浓度处理第10 天时增幅最大,较对照增加102%。木质素、黄酮和单宁酸在处理第5天时的含量最高,其中高浓度处理分别较对照增加113%, 75%和57%。用0.1 mmol/L茉莉酸诱导处理后,舞毒蛾2龄幼虫对叶片的消化率、转化率和利用率均有所降低, 取食处理后1, 5和10 d的青杨叶片的食物利用率较对照分别降低了29.4%, 27.6%和28.2%,且幼虫体重降低、发育历期延长。【结论】结果提示外源茉莉酸诱导青杨可对舞毒蛾产生明显的生化抗性,实践中可利用这种生化抗性防治舞毒蛾的危害。     

Thermal acclimation of leaf respiration but not photosynthesis in Populus deltoides x nigra

Dark respiration and photosynthesis were measured in leaves of poplar Populus deltoides x nigra ('Veronese') saplings to investigate the extent of respiratory and photosynthetic acclimation in pre-existing and newly emerged leaves to abrupt changes in air temperature. The saplings were grown at three temperature regimes and at high and low nitrogen availabilities. Rates of photosynthesis and dark respiration (R(d)) were measured at the initial temperature and the saplings were then transferred to a different temperature regime, where the plants remained for a second and third round of measurements on pre-existing and newly emerged leaves. Acclimation of photosynthesis was limited following transfer to warmer or cooler growing conditions. There was strong evidence of cold and warm acclimation of R(d) to growth temperature, but this was limited in pre-existing leaves. Full acclimation of R(d )was restricted to newly emerged leaves grown at the new growth temperature. These findings indicate that the extent of thermal acclimation differs significantly between photosynthesis and respiration. Importantly, pre-existing leaves in poplar were capable of some respiratory acclimation, but full acclimation was observed only in newly emerged leaves. The R(d)/A(max) ratio declined at higher growth temperatures, and nitrogen status of leaves had little impact on the degree of acclimation.     

花椒叶浸提液对土壤微生物数量和土壤酶活性的影响

通过用花椒叶浸提液浇灌盆栽花椒幼苗,研究浸提液对土壤酶和土壤微生物的影响.结果表明, 花椒叶浸提液使根际土中细菌、真菌和放线菌数量以及微生物总数均有不同程度的减少,根际土中真菌和放线菌的数量变化呈降-升-降-升的趋势.20、60和80 g·L^-1浓度的叶浸提液使非根际土中细菌的数量显著增加21.59%、107.55%和8.62%,而40 g·L-1浓度的叶浸提液则使非根际土中细菌数量显著降低22.56%.叶浸提液使根际土蛋白酶、蔗糖酶和酸性磷酸酶活性明显低于非根际土相应的酶活性,而过氧化氢酶和多酚氧化酶活性则显著升高.土壤的蛋白酶活性与蔗糖酶活性呈显著正相关,与土壤放线菌数量呈显著负相关;多酚氧化酶活性与蔗糖酶活性呈显著负相关,与细菌、真菌、放线菌以及微生物总数呈显著正相关;放线菌只与蛋白酶、多酚氧化酶、蔗糖酶3种酶活性及真菌呈显著相关,与过氧化氢酶、酸性磷酸酶以及细菌和微生物总数的相关性均不显著.     

外源茉莉酸诱导的青杨叶片保护性酶活性变化及其对舞毒蛾幼虫生长发育的影响

为探讨外源茉莉酸对青杨Populus cathayana的诱导抗性及其对舞毒蛾Lymantria dispar的影响, 室内对青杨扦插苗喷施不同浓度的茉莉酸（对照为0.17%丙酮）, 分别在喷施后1, 5和10 d采集叶片分析其保护性酶活性的变化, 并接舞毒蛾幼虫于青杨苗木上观测其长发育情况。结果表明： 0.1 和0.001 mmol/L两种浓度的茉莉酸（JA）处理均使青杨叶过氧化物酶（POD）、 多酚氧化酶（PPO）、 苯丙氨酸解氨酶（PAL）、 胰凝乳蛋白酶抑制剂（CI）和胰蛋白酶抑制剂（TI）活性较对照增加(P<0.05)。取食茉莉酸诱导的青杨苗木后, 舞毒蛾幼虫的发育历期延长, 体重降低。0.1 mmol/L茉莉酸诱导的青杨苗木5 d后接虫, 使舞毒蛾幼虫的发育历期显著延长, 较对照长8 d; 接虫21 d后称重时, 取食茉莉酸诱导的青杨叶片的幼虫体重较对照组降低了50%~100%, 该结果说明外源茉莉酸诱导青杨产生了对舞毒蛾明显的抗虫性。     

Some aspects of the morphology and distribution of Microsoriurn linguaeforrne (Polypodiaceae)

Microsorium linguaeforme is reported for the first time from India. Its rhizome is slender, much elongated with a highly dissected stele and leaf gaps in a single median dorsal row. The characteristic branching pattern is a modification of the common pattern in Polypodiaceae, resulting from the displacement of leaf-associated branches and the precocious development of the most basal secondary branch of each primary branch. The first two or three leaves of juvenile plants have no associated branches; thereafter, abaxially-originating traces t o solitary branches are progressively displaced so that the branch trace is close to the preceding leaf trace. Stomata of adult leaves are Copolomesoperigenous and of juvenile leaves Eupolomesoperigenous. The spores are monolete and with a smooth exine. Spore germination is of the Vitiariatype and prothallial development of the Kauliniatype. It is concluded that M. linguaeforme is closely allied to Leptochilus and is probably parental to it.     

Rapid alkalinization factors in poplar cell cultures. Peptide isolation,cDNA cloning,and differential expression in leaves and methyl jasmonate-treated cells

A family of peptides inducing rapid pH alkalinization in hybrid poplar (Populus trichocarpa x Populus deltoides) cell culture medium was isolated from hybrid poplar leaves. Five related approximately 5-kD peptides were purified by high-performance liquid chromatography and analyzed by matrix-assisted laser desorption ionization-mass spectrometry. The N-terminal sequence of one of the isolated peptides was very similar to a previously characterized peptide from tobacco (Nicotiana tabacum), rapid alkalinization factor (RALF), which causes a rapid increase in culture medium pH when added to tobacco cell cultures (G. Pearce, D.S. Moura, J. Stratmann, C.A. Ryan [2001] Proc Natl Acad Sci USA 98: 12843-12847). Two unique poplar RALF cDNAs (PtdRALF1 and PtdRALF2) were isolated from a poplar cDNA library and used to study RALF expression in poplar saplings and cultured poplar cells. Both genes were found to be expressed constitutively in poplar saplings and cultured cells. However, PtdRALF2 was expressed in leaves at very low levels, and its expression in suspension culture cells was transiently suppressed by methyl jasmonate (MeJa). Although the function of these novel peptides remains enigmatic, our experiments suggest their role may be developmental rather than stress related. Overall, our study confirms the presence of active RALF peptides in other plants, and provides new data on the complexity of the RALF gene family in poplar.     

Effects of Alternating Night Temperature and Cytokinin Application on the Activity of Enzymes of Sucrose Metabolism in Leaves, Young Shoots and Petals of Roses

Different aspects of growth and development of rose plants grownat constant high or low night temperatures in association withthe activity of enzymes of sucrose metabolism and effects ofcytokinin application were previously described (Khayat andZieslin 1987a). It has also been shown that growth and floweringwere affected when rose plants were grown at night temperaturesof 18?C alternating every 2 h with 12?C. Data on the effectsof alternating night-temperature and application of cytokininon the activity of those enzymes in various tissues of roseplants, in comparison to average activity at constant high andlow temperatures are described in the present report. The resultsshowed that the activities of acid invertase in leaves and youngshoots and of sucrose synthase in petals of rose plants grownat alternating night temperatures were similar to the averageactivity. The alternating temperatures reduced the activityof acid invertase in petals, slightly promoted the activityof sucrose-phosphate synthase in the leaves and enhanced theactivity of sucrose synthase in young shoots. Application ofbenzyl adenine to an axillary bud in plants grown at alternatingtemperature caused a 2-fold increase in the activity of sucrosesynthase and reduced the activity of acid invertase in youngshoots and promoted the activity of sucrose-phosphate synthaseand acidic invertase in leaves as compared with the averageactivity in plants treated with benzyladenine and grown at constanthigh and low temperatures. The relevance of these phenomena to flower formation in rosesis discussed. (Received November 6, 1989; Accepted June 16, 1990)     

A mechanical analysis of the relationship between free oscillations of Pinus pinaster Ait. saplings and their aerial architecture

The aim of this study was to investigate the influence of aerial architecture on the dynamic characteristics of young maritime pines (Pinus pinaster Ait.) using a mechanistic approach. For this purpose, three 4-year-old saplings with prominent differences in their branching patterns were submitted to free oscillation tests. The tests were carried out with different methods and directions of mechanical loading in order to initiate the movement of each sapling. The oscillations of the different architectural elements, i.e. stem and branches of different topological order, were measured with inclinometers and strain gauges fixed to saplings. Successive pruning of the architectural elements was carried out to evaluate their relative influence on the dynamic characteristics of the trees. The aerial systems were digitized before the mechanical tests in order to use 3D visualization techniques and to make architectural analyses of the crown structure. Two distinct modes of deformation were detected during free oscillations. The natural swaying frequency ranged from 0.6-0.8 Hz for the saplings tested at the same period of the year. The frequency variations were partly explained by the morphological differences of the experimental subjects. The motions of the axes were found to depend on their topology, i.e. the movement of the axes of a given branching order was forced by the movement of their respective bearing axis. The axes of third branching order had a significant and negative effect on the damping of the natural deformation mode. Results point out the major role played by foliage, qualitatively and quantitatively, on the damping of tree motions and on coupling the motions of the crown components.     

CsRAV1 induces sylleptic branching in hybrid poplar

? Sylleptic branching in trees may increase significantly branch number, leaf area and the general growth of the tree, particularly in its early years. Although this is a very important trait, so far little is known about the genes that control this process. ? This article characterizes the Castanea sativa RAV1 gene, homologous to Arabidopsis TEM genes, by analyzing its circadian behavior and examining its winter expression in chestnut stems and buds. Transgenic hybrid poplars over-expressing CsRAV1 or showing RNA interference down-regulated PtaRAV1 and PtaRAV2 expression were produced and analyzed. ? Over-expression of the CsRAV1 gene induces the early formation of sylleptic branches in hybrid poplar plantlets during the same growing season in which the lateral buds form. Only minor growth differences and no changes in wood anatomy are produced. ? The possibility of generating trees with a greater biomass by manipulating the CsRAV1 gene makes CsRAV1 transgenic plants promising candidates for bioenergy production.     

Activities of Sucrose Synthase and Acid Invertase in Pea Seedling Organs

The organ topography of sucrose synthase and soluble acid invertase in pea seedlings at heterotrophic stage (3–14 days) was studied. Sucrose synthase was most active in the roots, with the highest activity on the 6–8th days. In the leaves, its activity decreased from day 3 to day 14. In the stems, sucrose synthase activity was at an invariantly low level. The patterns of sucrose synthase activity in etiolated and green plants were similar. As distinct from sucrose synthase, invertase activity was the highest in the stem, especially in etiolated plants. The peak of its activity was observed on the 6-8th days. In the leaves, invertase activity was lower but its pattern was the same. In the roots, acid invertase activity decreased from the 3rd day and did not depend on illumination. The conclusion is that differences in sucrose synthase and acid invertase activities in roots, leaves, and stem are determined by differences in the import of hydrolytic products of stored compound from the cotyledons as well as by different demands of these organs for these products for the processes of organ expansion and for the maintenance of organ metabolism.     

Inter- and intraspecific relationships between shade tolerance and shade avoidance in temperate trees

Patterns of physiological and architectural adaptation and acclimation to decreasing light availability were investigated along a light gradient for saplings of 12 common species of temperate deciduous trees in southeastern Ontario, Canada. Physiological adaptation and acclimation (shade tolerance physiology) were quantified at the leaf level by measuring leaf mass per unit area (LMA), dark respiration per unit leaf nitrogen, chlorophyll per unit leaf nitrogen and the chlorophyll a:b ratio for the newest fully expanded leaf on the leader. Architectural adaptation and acclimation (shade avoidance) were quantified by measuring branching intensity and side shoot:main shoot length ratios for the most recent three years of growth on the leader and selected side branches. Within species, increases in LMA, chlorophyll a:b ratio and respiration per unit nitrogen and decreases in chlorophyll per unit nitrogen indicated that shade tolerance physiology generally increased with decreasing canopy openness. Increases in the branching intensity and side shoot:main shoot ratios of the leader and side branches indicated that shade avoidance also increased with decreasing canopy openness for the majority of species; however, in some species, stem bending under deep shade resulted in lateral growth. Interspecific variation in shade tolerance physiology was minimal when species were compared under equal amounts of canopy openness. In contrast, interspecific variation in shade avoidance variables was relatively high under equal canopy openness, with saplings of shade tolerant canopy species exhibiting higher shade avoidance than saplings of shade tolerant understory species.     

Involvement of putative chemical wound signals in the induction of phenolic metabolism in wounded lettuce

Cutting leaves of Romaine lettuce ( Lactuca sativa L. cv. Longifolia) produces a wound signal that induces the synthesis of phenylalanine ammonia lyase (PAL, EC 4.3.1.5) and the accumulation of phenolic compounds in cells up to 2 cm from the site of injury, and tissue browning near the site of injury. The response of leaves within a head of Romaine lettuce to putative chemical wound signals [abscisic acid (ABA), jasmonate (JA) and methyl jasmonate (MeJA)] differed significantly with leaf age. Exposure of harvested heads of lettuce to ABA, JA, MeJA, or salicylic acid (SA) did not induce changes in PAL activity, the concentration of phenolic compounds or browning in mature leaf tissue that was similar to the level induced by wounding. Methyl jasmonate applied as vapour (10, 100 or 1000 µl kg⁻¹ FW), or as an aqueous spray or dip (0.01–100 µ M ) at 5 or 10°C did not produce an effect on PAL activity or browning that differed significantly from the untreated controls. In contrast, JA, MeJA and SA did induce elevated levels of PAL activity in younger leaves. However, the levels induced were far lower than those induced by wounding. Wound induced phenolic metabolism in mature leaves appears to be induced by different signals than those functioning in young leaves.