DEVELOPMENTAL ANATOMY OF JUVENILE AND ADULT SHOOTS OF MARCGRAVIA RECTIFOLIA L

Marcgravia rectifolia L. is a dimorphic vine having distinct juvenile and adult shoots. The juvenile shoot is a climber characterized by an orthotropic growth habit, a flattened stem, adventitious roots, and ovate leaves. The adult shoot, on the other hand, possesses a plagiotropic growth habit, has a cylindrical stem, few or no adventitious roots, and lanceolate leaves. Both phases have distichous phyllotaxy, however the plastochron is shorter for the adult phase than for the juvenile. Internode elongation occurs earlier for adult shoots than for juvenile shoots. Cytological analyses show the flattened stem of the juvenile results from differential production of cells, especially in the pith region. On the other hand, internodes of the adult phase are longer than juvenile internodes, a result of more cells produced rather than longer cells. In juvenile stems a perivascular band of elongated fibers develops, while in adult stems this band consists of brachyosclereids. Both phases undergo secondary growth and have non-storied cambia. Cambial activity begins in the 6th internode of each phase. As secondary growth proceeds, the adult stem produces much more xylem than juvenile stems of the same age. Adventitious roots produced in the juvenile stem are located in vertical rows at the “corners” of flattened stems and are attachment structures aiding the climbing habit of this vine. Phase changes occur regularly in this species. The juvenile phase usually transforms into the adult, however the adult phase can spontaneously revert back into the juvenile phase. The anatomical features and the phase changes are discussed and compared to Hedera helix, a vine whose phase changes have been studied in some detail. It is suggested that the anatomical features of Marcgravia rectifolia L. including its phase changes, may provide an alternative system to study physiological changes similar to those done with Hedera helix.     

THE BEHAVIOR OF TISSUE CULTURES FROM ENGLISH AND ALGERIAN IVY IN DIFFERENT GROWTH PHASES

In both the English and Algerian ivies, Hedera helix L. and Hedera canariensis Willd., leaf dimorphism of the juvenile and the mature, fruiting growth phases is pronounced, the former being a vine with lobed leaves and the latter a shrubby, upright form with entire leaves. Tissue cultures of English ivy started from stems of the two growth phases on the same plant consistently behaved differently, those from the juvenile stage invariably having the higher proliferation rate and larger cells. These differences were maintained consistently in monthly transfers over a period of two years. No medium was found which would support the growth of tissue subcultures of the adult stage of Algerian ivy, but all growth phases of the English ivy were cultured freely in a modified White's medium with additions of coconut water, naphthaleneacetic acid, enzymatic casein hydrolysate, and inositol. Cultures from individual open-pollinated seedlings of both species varied greatly in proliferation rate but were usually high.     

COMPARATIVE DEVELOPMENTAL ANATOMY OF SHOOTS OF JUVENILE AND ADULT HEDERA HELIX

Apices of adult Hedera helix have a larger meristematic area, composed of smaller cells, than those of the juvenile shoots. In shoot tips of juvenile plants, cell divisions in the subapical area occur over a longer portion of the shoot, and cell division is continued for a longer period of time. These features are reminiscent of GA-induced changes in other plants. It is suggested that in experiments designed to shift Hedera from the juvenile to the adult form, consideration should be given to agents which affect both rates of cell division and distribution of dividing cells in the young shoot.     

Proteins and chlorophyllase activity in Hedera helix L. with green or variegated leaves

Abstract

Chlorophyllase activity and photosynthetic membrane proteins of two lots of Hedera helix were studied. Both the HV variety with variegated leaves and HC variety with green leaves were observed. Proteins from green, white and white-green portions of leaves differing in age were compared using dodecyl-sulfate-polyacrylamide gel electrophoresis. The white or white-green regions of Hedera helix var. aureomarginata (HV) did not differ significantly in chlorophyllase activity.     

Photosynthesis in leaves of the juvenile and adult phase of ivy (Hedera helix)

Abstract Photosynthetic and anatomical parameters of leaves from the juvenile and adult part of an ivy plant (Hedera helix L.) have been determined and compared with each other. Light-saturated net photosynthesis (per unit leaf area) was about 1.5 times higher in adult leaves than in juvenile ones. The lower photosynthetic capacity of juvenile leaves was caused by a lower stomatal and especially a lower residual conductance to the CO₂-transfer. This corresponds with anatomical features of the leaves, i.e. lower stomatal frequency, fewer chloroplasts per cell, and – most important – thinner leaves, as well as with a less efficient photosynthetic apparatus measured as Hill reaction of isolated broken chloroplasts and activity of ribulose bisphosphate carboxylase. No differences in the respiration in light (relative to net photosynthesis) and in the CO₂-compensation concentration could be detected between the two leaf types. These observed anatomical and photosynthetic parameters of the juvenile and adult ivy leaves resemble those reported for shade and sun leaves, respectively, although the leaves investigated originated from the same light environment.     

Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix)

Hoflacher, H. and Bauer, H. 1982. Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix). – Physiol. Plant. 56: 177–182. Light acclimation was investigated during the juvenile and adult life phases of the whole-plant-development in Hedera helix L. For this purpose, cuttings of the juvenile and adult parts of one single parent plant were grown under low-light (PAR 30–50 μmol photons m^?2 s^?1) and high-light (PAR 300–500 μmol m^?2 s^?1) conditions: CO₂ exchange, chloroplast functions, and specific anatomy of fully developed leaves differentiated under these conditions were determined. In juvenile plants the leaves formed under low and high light had light-saturated rates of net photosynthesis of 6.5 and 11.1 mg CO₂ (dm leaf area)^?2 h^?1, respectively. In adult plants the rates were 9.4 and 22.2 mg dm^?2 h^?1, indicating a more pronounced capacity for acclimation to strong light in the adult life phase. Higher photosynthetic capacities were accompanied by higher conductances for the CO₂ transfer through the stomata, leading to almost the same CO₂ concentration in the intercellular spaces. Thus, stomatal conductances were not primarily responsible for the different photo-synthetic capacities. The higher rates in adult and high-light grown leaves were mainly the result of formation of thicker leaves with more chloroplasts per unit leaf area. Expressed per chloroplast, the photosynthetic capacity, the Hill reaction, and the activity of ribulose bisphosphate carboxylase were almost identical in plants grown in low-light and high-light. Measurements of photosynthetic capacity and thickness of leaves of Hedera sampled from field habitats with contrasting light regimes confirm the results of growth chamber studies. It is, therefore, concluded that both life phases of Hedera are capable of acclimating to strong light, but that during the juvenile phase this capacity is not fully developed.     

Frost hardiness in the juvenile and adult life phase of ivy (Hedera helix L.)

The common ivy (Hedera helix L.) remains juvenileat its northern, eastern and altitudinal distribution limits although juvenileparts are largely killed by severe frost spells. In order to explain thisdiscrepancy we investigated the seasonal course of frost resistance in variousorgans of juvenile and adult parts of the same H. helixplants. Maximum frost resistance of leaves (LT₅₀–25°C) and axis (xylem parenchyma:LT₅₀ –29°C; cambium: LT₅₀–35°C) was quite the same in juvenile and adultparts. Thus, H. helix is able to acquire full frostresistance in its juvenile phase. However, hardening of leaves was slower anddehardening of axis was faster in juvenile parts. Leaves of juvenile partsremain 2 to 4 K less resistant than those of adult parts untilattaining the maximum resistance. This explains why mainly leaves of thejuvenile parts were damaged following severe frost episodes with temperaturesbelow –20°C. The occurrence of H.helix in its juvenile phase at the frost-caused distribution limitsmay be explained as follows: Leaves of juvenile plants may occasionally bekilled by severe frosts, but regeneration from dormant eyes enables survival.Loss of leaves may impede the change to the adult phase, but even if the plantsbecame adult frost killing of rest buds (2 to 3 K less resistantthan leaves) would induce rejuvenation.     

Ribosomal RNA Gene Redundancy in Juvenile and Mature Ivy (Hedera helix)

Two-fold variation was found in rRNA gene redundancy in differentpopulations of ivy (Hedera helix). No consistent differencein rRNA gene number was observed between the juvenile and matureforms of the plant. Small significant differences between therRNA gene content of mature and juvenile forms and also betweenundeveloped buds and expanded leaves were found but these werein different directions in different populations.     

PLASTOCHRON INDICES FOR JUVENILE AND MATURE FORMS OF HEDERA HELIX L. (ARALIACEAE)

English ivy (Hedera helix) plants were assessed for the applicability of the plastochron index (PI). Juvenile ivy satisfied all requirements for the use of the PI and showed a plastochron of 4.23 days. Mature ivy grown under long day conditions flowered after 11–12 leaves. Two distinct groups of leaves were produced with different plastochrons (0.83 and 3.2 days, respectively) and leaf morphologies. Long-day-grown ivy did not satisfy the requirements for the use of the PI. Short-day-grown mature ivy continued production of leaves beyond the 12th leaf. Vegetative growth was perpetuated for at least 25 plastochrons. By 19 plastochrons (ca. 41 days after budbreak) a linear PI vs. time relationship was established with a plastochron of 3.16 days. This newly acquired ability to maintain vegetative growth in mature ivy plants may allow a direct comparison with the vegetative indeterminant juvenile in order to assess possible anatomical factors responsible for phase stability and phase change using the PI as a basis for comparison.     

Phase Change in Hedera helix L: I. GIBBERELLIN-LIKE SUBSTANCES IN THE TWO GROWTH PHASES

Gibberellin-like substances were extracted from all parts ofthe juvenile and adult growth phases of Hedera helix. A numberof extraction techniques were employed and estimatos of theamount of gibberellin-like activity present were made usingthree bioassays. Gibberellin like substances were present inapproximately equal amounts in leaves at the two growth phasesand appeared to be localized, in the main, in chloroplasts.Apical ‘buds’ of the juvenile phase were shown tocontain higher levels of gibberellin-like substances than theiradult counterparts. Roots, which under normal circumstancesare associated only with the juvenile phase, contained highlevels of extractable gibberellin-like substances.     

GROWTH AND HABITUATION IN TISSUE CULTURES OF ENGLISH IVY,HEDERA HELIX

Tissue cultures from both juvenile and adult stems of English ivy, Hedera helix L., were established in White's medium supplemented by coconut water and auxin (usually naphthalene acetic acid). With repeated transfers, cultures were habituated in less than a year to grow well without coconut water by using an auxin and kinetin. Cultures from juvenile seedlings were less demanding in requirements for growth. In all types of cultures occasionally small areas of rapidly growing cells were noticed. These when isolated gave rise to rapidly growing cultures with many cells of unusual appearance. Abnormally long cells and chain-type cells were abundant. When 0.1 mg/ liter of kinetin was added to the medium, these cells grew well without auxin and coconut water.     

Responses of ivy (Hedera helix L.) to combinations of gibberellic acid,paclobutrazol and abscisic acid

A reduction in concentration of gibberellins has been implicated in the phase change from juvenile to mature forms of ivy (Hedera helix L.). Attempts were made to increase the effective internal concentration of gibberellins by exogenous application of GA₃, and to decrease them by various applications of abscisic acid (ABA) and paclobutrazol (PP333), alone or in combination with GA₃. ABA and GA₃ were fed directly into the xylem of ivy plants by a wick system (a less drastic procedure than the defoliation or decapitation used by earlier workers) whereas PP333 was applied as a soil drench.Mature ivy responded to the application of GA₃ by reversion to the juvenile form, although this reversion was incomplete with respect to leaf lobing and red (anthocyanin) pigmentation and could occur spontaneously without the application of GA₃. Contrary to expectation, applications of ABA and PP333 caused the stimulation of growth in juvenile ivy. No adult characteristics were induced. As similar concentrations of ABA and PP333 produced severe retardation of growth (which could be alleviated by the application of GA₃) in other species, it is suggested that ivy may be an unsuitable model system for the investigation of phase change in woody plants.     

Behavioural responses of the vine weevil, Otiorhynchus sulcatus, to semiochemicals from conspecifics, Otiorhynchus salicicola, and host plants

The vine weevil Otiorhynchus sulcatus is a parthenogenetic reproducing species which forages for suitable host plants at night, but is found congregated in dark places during the day. Frass of this weevil species is suspected to contain attractive compounds that are host‐plant related. Using a still‐air olfactometer, we tested adult vine weevils at night for their behavioural response to odours from conspecifics, feeding on a mixture of spindle tree (Euonymus fortunei) and yew (Taxus baccata), and to a sexually reproducing related species (Otiorhynchus salicicola), feeding on a mixture of ivy (Hedera helix) and cherry laurel (Prunus laurocerasus). Their attraction to conspecifics and O. salicicola appeared to be related to frass production. Freshly collected frass from O. sulcatus and from O. salicicola males and females was attractive. Prunus laurocerasus and H. helix have not been observed to be hosts of the vine weevil in the field. However, our tests showed that the vine weevil was attracted to mechanically damaged leaves of both plant species, whereas undamaged leaves were not attractive. Only undamaged young unfolding leaves of H. helix were also attractive. The attraction to odours from mechanically damaged host and non‐host plants suggested the involvement of compounds that are commonly found in many plant species. The involvement of plant compounds and/or aggregation pheromones in attraction to frass of the vine weevil and frass of the related weevil species O. salicicola is discussed.     

Phase Change in Hedera helix L: II. THE POSSIBLE BOLE OF ROOTS AS A SOURCE OF SHOOT GIBBERELLIN-LIKE SUBSTANCES

When synthesis was estimated by the agar diffusion techniqueboth basal and lateral adventitious roots of Hedera helix L.in the juvenile growth phase were shown to synthesize gibberellin-likesubstances. Seedlings and cuttings from juvenile ivy could begrown in water culture for several weeks. When roots were excisedfrom these plants shoot growth was reduced in comparison withthat of intact plants. The stems, apices, and leaves of derootedseedlings and cuttings contained lower levels of extractablegibberellin-like substances than comparable organs of intactplants. The major zone of gibberellin-like activity in intactplants co-chromatographed with gibberellins A₁ and A₃. Whenthese gibberellins were applied to plants grown in culture theywere found to promote growth of intact but not derooted plants.These findings are discussed in relation to the role of rootfactors and particularly gibberellins in phase change.     

Phase Change in Hedera helix: Induction of the Mature to Juvenile Phase Change by Gibberellin A3

A sensitive and reproducible method to obtain GA₃ induced morphological reversion of mature Hedera helix to the juvenile form has been developed. Dose response experiments indicate that GA₃ stimulates reversion over a 50–100 fold range with a half maximal response at approximately 0.5 μg GA₃ per plant. The individual characteristics involved in phase change revert to the juvenile form in a sequential manner as GA₃ dose is increased. Variations in light intensity from 1.2–3.6 × 10⁴ lux and temperature from 15 to 26°C do not affect this hormonal response. Other growth regulators including indoleacetic acid, kinetin, abscisic acid and (2-chloroethyl)phosphonic acid (Ethephon) are inactive but other gibberellins (GA₁ and a mixture of A₄–A₇) are active in stimulating reversion. Therefore, the response is specific for gibberellins as a class of hormones but non-specific for a particular form of gibberellin. The significance of this response in relation to juvenility in woody plants is discussed.     

The effect of light on the Og of plant tissues. I

Summary The increment of O_g in the guard cells in response to light as reported by previous investigators was found to be true qualitatively but not quantitatively.A special device was constructed, which permitted the elimination of the heat factor when the leaves were irradiated.In experiments 1 and 2 the relative values of the bell and the device were determined.In experiments 3 and 4 the relative effects of light were studied on younger and older leaves.In experiments 5 and 6 the effects were studied on old and young leaves ofHedera helix. This plant serves as a standard for comparisons of results obtained by subjecting plants to various factors of the environment.In experiments 7 and 8 the effect of light on young, middle-aged and old leaves ofHedera helix, when water was supplied, in one case but not in the other were studied. In experiment 8 the same leaf was treated in the device and then in the dark chamber, and once more in the light. The results are particularly enlightening.In the supplementary experiments, it was verified that the phenomenon of an increment of the guard cells and a decrement of the spongy parenchyma in O_g, when irradiated, is probably general.     

REGULATORY ROLE OF INDOLE-3-ACETIC ACID AND GIBBERELLIC ACID IN VEGETATIVE DEVELOPMENT OF XANTHIUM PENNSYLVANICUM

A single application of gibberellic acid to young internodes significantly accelerated the rate of internode growth and the rate of leaf production in shoots of Xanthium pennsylvanicum Wallr. The average duration of one plastochron in treated plants was reduced to 43% of control levels. Gibberellic acid also had a pronounced morphogenetic effect on leaves so that the area and leaf length of treated plants were both significantly reduced. Depending upon concentration, auxin had both inhibitory and promotive effects on Xanthium shoots. Indole-3-acetic acid markedly altered the response of the gibberellic acid-treated internodes and those located above and below the site of application. In addition, high auxin concentrations induced the formation of adventitious roots in treated internodes. Auxin also brought about significant reductions in the length and area of leaves developed under the influence of this hormone.     

Polyamines and Adventitious Root Formation in the Juvenile and Mature Phase of English Ivy

The involvement of polyamines during adventitious root formationwas evaluated using a de-bladed petiole rooting assay for theeasy-to-root juvenile and difficult-to-root mature phase ofEnglish ivy (Hedera helix L.). Auxin (NAA 0.1 mM) stimulatedroot formation in juvenile phase cuttings, but failed to promoterooting in the mature phase. The addition of putrescine, spermineor spennidine (1.0 mM) with or without NAA (0.1 mM) did notaffect the rooting response in either the juvenile or maturephase cuttings. There was a significant increase in endogenouslevels of putrescine and spermidine in NAA-treated cuttings,but the only significant difference between the root formingjuvenile and the non-root forming mature phase cuttings wasan increase in putrescine levels. In NAA-treated juvenile cuttings,the polyamine biosynthesis inhibitor DFMA (1.0 mM) promotedroot formation from 9.2 to 14.5 roots per cutting, while DFMO(1.0 mM) reduced root formation from 9.1 to 1.4 roots per cutting.The promotion of rooting by DFMA was completely reversed byputrescine (1.0 mM), but putrescine, spermine or spermidine(1.0 mM) could not reverse the inhibitory effect of DFMO. NeitherDFMA nor DFMO promoted root formation in mature phase cuttings.DFMA was also added to NAA-treated juvenile petioles at variousstages during the root formation process. DFMA promoted rootingwhen applied during the early stages of root induction (0–3d), but became inhibitory to root formation when applied duringthe organization (6–9 d) or root elongation stages (9–12d). Key words: Hedera helix, organogenesis, root initiation, polyamines, DFMA, DFMO     

Stomatal Responses to Light and Drought Stress in Variegated Leaves of Hedera helix

Direct and indirect mechanisms underlying the light response of stomata were studied in variegated leaves of the juvenile phase of Hedera helix L. Dose response curves of leaf conductance were measured with blue and red light in leaves kept in normal or in an inverted position. In the green portions of the leaves, the sensitivity to blue light was nearly 100 times higher than that to red light. No response to red light was observed in the white portions of the leaves up to 90 micromoles per square meter per second. Red light indirectly affected leaf conductance while blue light had a direct effect. Leaf conductance was found to be more sensitive to drought stress and showed a more persistent aftereffect in the white portions of the leaves. A differential effect of drought stress on the responses to blue and red light was also observed.     

Proserpinaca: photoperiodic and chemical differentiation of leaf development and flowering

Proserpinaca palustris L. produced juvenile leaves on 8-hour photoperiods, adult leaves on 12-hour photoperiods, and adult leaves and flowers on 14-hour photoperiods. Treatment of plants growing on 8- and 14-hour photoperiods with gibberellic acid caused stem elongation and inhibited flowering. The treated plants on 8-hour photoperiods produced adult-like leaves.