Developmental Genetics and Morphological Evolution of Flowering Plants, Especially Bladderworts (Utricularia): Fuzzy Arberian Morphology Complements Classical Morphology

This review compares new developmental models on flowering andother vascular plants with evolutionary hypotheses formulatedby Agnes Arber (1879–1960) and like-minded botanists.Special emphasis is laid on philosophical basics such as perspectivism,pluralism about evolutionary modelling, continuum way of thinking,and fuzzy logic. Arber's perspective is best labelled as F uzzyA rberian M orphology (FAM Approach). Its proponents (‘FAMmers’)treat structural categories (e.g. ‘roots’, ‘shoots’,‘stems’, ‘leaves’, ‘stipules’)in vascular plants as concepts with fuzzy borderlines allowingintermediates (including transitional forms, developmental mosaics).The FAM Approach complements Cla ssical Plant M orphology (ClaMApproach), which is the traditional approach in botany. ClaMproponents (‘ClaMmers’) postulate that the structuralcategories of vascular plants are regarded as concepts withclear-cut borderlines and without intermediates. However, duringthe evolution of vascular plants, the root-shoot distinctionand the stem-leaf distinction have become blurred several timesdue to developmental changes, resulting in organs with uniquecombinations of features. This happened, for example, in thebladderworts (Utricularia, Lentibulariaceae). When focusingon the ‘leaf’, the FAM Approach is identical toArber's ‘partial-shoot theory of the leaf’ and Sinha's‘leaf shoot continuum model’. A compound leaf canrepeat the developmental pathway of the whole shoot, at leastto some degree. For example, compound leaves of Chisocheton(Meliaceae)with indeterminate apical growth and three-dimensional branchingmay be seen as developmental mosaics sharing some growth processeswith whole shoots! We focus here on the FAM Approach becausethis perspective is especially promising for developmental geneticistsstudying flowering and other vascular plants. Copyright 2001Annals of Botany Company Review, body plan, developmental mosaics, leaf development, history of botany, homeosis, homeotic genes, Lentibulariaceae, morphological evolution, process morphology, stipules, Utricularia, flowering plants     

Topophysis affects the Potential of Axillary Bud Growth, Fresh Biomass Accumulation and Specific Fresh Weight in Single-stem Roses (Rosa hybrida L.)

Topophysis, the effect on growth and differentiation of positionof axillary buds along the shoot, was studied by propagatingfive-leaflet-leaf single-node cuttings which were excised fromseven stem positions and grown as single stemmed plants. InRosahybrida ‘Korokis’ Kiss®, ‘Tanettahn’Manhattan Blue®, and ‘Sweet Promise’ Sonia®,following release of the buds from apical dominance by excision,morphogenetic development was studied until anthesis. The timefrom excision/planting until onset of bud growth, visible flowerbud appearance, and anthesis was generally shorter in plantsoriginating from apical bud positions than from basipetal positions.Topophysis mainly affected the onset of axillary bud growth;the earliest growth and development was found in cuttings fromthe second uppermost node position. This node tended to havethe lowest plastochron value, which indicated the existenceof a transition between sylleptic and proleptic buds. Stem lengthat visible flower bud and at anthesis generally increased asthe cutting position changed basipetally until the second lowestposition, and the number of five-leaflet-leaves at anthesisand the total number of nodes generally increased basipetally.For internode length, growth rate, and fresh biomass efficiencythe cuttings taken from the uppermost and lowermost positionsgenerally had significantly lower values than cuttings fromall medial positions. At anthesis, plants originating from cuttingsexcised from lower medial positions generally had a higher freshweight, greater flower stem diameter, and a significantly higherspecific fresh weight than those plants originating from apicalor basal positions. Among the cultivars, Sonia was the mostefficient in increasing fresh biomass and had the highest growthrate, whereas Manhattan Blue possessed the highest specificfresh weight, indicating a higher plant quality. It is suggestedthat topophysis inRosa is an independent phenomenon intrinsicto the axillary bud. apical dominance; axillary bud growth; fresh biomass accumulation; cut rose; flowering; Rosaceae; Rosa hybrida L.; rose; shoot growth; single-stem roses; specific fresh weight; topophysis; quality     

Narrow-leafed Lupins with Restricted Branching

Crop phenology is one of the most important characters influencingproductivity in a given environment. Narrow-leafed lupin (Lupinusangustifolius L.) is a major grain legume crop in southern Australiawith general phenological adaptation to this Mediterranean-typeenvironment. However, it is an indeterminate crop with severalassociated limitations to productivity, such as overlappingvegetative and reproductive growth, late grain filling and sometimesexcessive vegetative growth. Here we studied two novel typesof narrow-leafed lupin with restricted branching, which mightbe useful for overcoming these problems. These restricted branchinglupins arose spontaneously within a breeding population, inthe case of ‘Tallerack’, and within a farmer's cropin the case of ‘ Hurst’ and we compared them withthe ‘Merrit’, which is widely grown and has thenormal indeterminate branching habit. The morphology and developmentof the main shoot of these genotypes were similar. However,‘Hurst’ had much larger leaves. There were alsostriking differences in the lateral branches of the restrictedbranching types; they had fewer leaves than ‘Merrit’and flowered earlier. These differences were most marked in‘ Hurst’, where the upper main stem branches werereduced to a single floret in the axil of main stem leaves,and these flowers often exhibited abnormal morphology. Copyright2000 Annals of Botany Company Lupinus angustifolius L., narrow-leafed lupin, adaptation, development, morphology, branching, leaves, mutant, plastochron, phyllochron, floral initiation, flowering.     

Growth and Dry-weight Distribution in Callistephus chinensis as Influenced by Lighting Treatment

Plants of two cultivars of Callistephus chinensis (Queen ofthe Market and Johannistag) were grown in 8 h of daylight perday with one of the following treatments given during the 16h dark period: (a) darkness—‘uninterrupted night’,(b) I h of light in the middle of the dark period—a ‘nightbreak’, (c) I min of light in every hour of the dark period—‘cycliclighting’, (d) light throughout—‘continuouslight’. The plants receiving uninterrupted dark periods remained compactand rosetted in habit with small leaves, while leaf expansion,stem extension, and flower initiation were promoted in all threeillumination treatments (b, c, d). Although these three treatmentsproduced similar increases in leaf area, continuous light wasthe most effective for the promotion of both stem growth andflower initiation while cyclic lighting was generally more effectivethan a I-h night break. Continuous light also caused more dry matter to be divertedto stems at any given vegetative dry weight and it was shownthat the stem weight ratio of both varieties was correlatedwith stem length.     

Photothermal Time for Flowering in Lentils (Lens culinaris) and the Analysis of Potential Vernalization Responses

Two cultivars of lentils, Laird and Precoz, were subjected to18 potentially vernalizing treatments, comprising constant temperaturesof 1, 5 or 9 °C in factorial combination with photoperiodsof 8 or 16 h for 10, 30 or 60 d. These seeds or seedlings, togetherwith non-vernalized seeds (as controls), were then transferredto four different growing regimes (‘day’/‘night’temperatures of 18/5 °C or 24/13 °C, factorially combinedwith photoperiods of 11 or 16 h). Variation in the number ofdays from sowing to first flower (f) in the growing regimesfor the controls conformed to the equation I/f = a+b+cP, whereis mean temperature (°C), P is photoperiod (h) and a, band c are genotype-specific constants. Accordingly, when theenvironment varies during development, the photothermal timerequired to flower in day-degrees (°C d) is given by 1/babove a base temperature defined as —(a+cP)/b. Most variationin time to flower could be accounted for by the photothermaltime accumulated in the two successive environments. Therefore,there was no evidence of a specific low-temperature vernalizationresponse in either cultivar. Neither was there evidence of ‘short-day’vernalization, i.e. advancement of flowering resulting frompreliminary short-day treatments. A potential error inherentin the predictive model described arises because it ignoresthe presence of a pre-inductive, photoperiod-insensitive phase;but agro-ecological considerations suggest that this error maynot be important in practice. Lens culinaris, lentil, flowering, photoperiodism, vernalization, photothermal time, screening germplasm     

Influence of Flooding on Net CO2Assimilation, Growth and Stem Anatomy of Annona Species

A series of experiments was conducted to assess net CO₂assimilationand growth responses to waterlogging of grafted and seedlingtrees in the genus Annona. Seedlings of A. glabra, A. muricataandA. squamosa L., and scions of ‘Gefner’ atemoya(A. squamosaxA. cherimola Mill.), ‘49-11’ (‘Gefner’atemoyaxA. reticulata L.), ‘4-5’ (‘Priestley’atemoyaxA. reticulata), A. reticulata grafted onto either A.glabra, A. reticulata orA. squamosa rootstocks were floodedfor up to 60 d. Soil anaerobiosis occurred on the third dayof flooding. Seedlings ofA. glabra and A. muricata, and thescions ‘49-11’, ‘Gefner’ atemoya, andA. reticulata grafted onto A. glabra rootstock were consideredflood tolerant based on their ability to survive and grow inflooded conditions. Scions of the normally flood-sensitive A.reticulata, ‘Gefner’ atemoya, and ‘49-11’tolerated root waterlogging when grafted onto the flood-tolerantspecies, A. glabra. In contrast, flooding of A. squamosa seedlingsand rootstocks, and A. reticulata rootstocks greatly reducedgrowth and net CO₂assimilation rates, and resulted in 20–80%tree mortality. Stem anatomical responses to long-term flooding(12 continuous months) were assessed in seedlings of A. glabraand A. muricata, and trees of ‘49-11’ grafted ontoA. glabra. Flooded trees developed hypertrophied stem lenticels,particularly in A. glabra, and enlarged xylem cells resultingin thicker stems with reduced xylem density. Flooding did notincrease air spaces in pre-existing xylem near the pith or inxylem tissue that was formed during flooding. Thus, flood tolerancedid not involve aerenchyma formation in the stem. Copyright1999 Annals of Botany Company Flood tolerance, net CO₂assimilation, photosynthesis, stem anatomy, shoot growth, anaerobiosis, Annonaceae.     

Relative Sensitivity and Tolerance of some Gladiolus Cultivars to Sulphur Dioxide

Five Gladiolus cultivars, namely ‘Aldebaran’, ‘BrightEye’, ‘Illusion’, ‘Manisha’ and‘Manmohan’, were exposed to 1 and 2 µg l^–1sulphur dioxide to test their relative-sensitivity toleranceto the pollutant Plants were fumigated experimentally for 2h daily Foliar injury symptoms were observed first in ‘Manisha’followed by ‘Aldebaran’ and ‘Illusion’at the higher dose Photosynthetic pigments and leaf extractpH were significantly decreased, particularly in ‘Manisha’and ‘Illusion’ Overall disturbances in the plantmetabolism due to SO₂ treatment led to retarded growth of plants,as evident from decreased shoot length and phytomass valuesThe order of sensitivity of the five Gladiolus cultivars toSO₂ was as follows, with the greatest first Manisha, Illusion,Aldebaran, Bright Eye, Manmohan Cultivars, Gladiolus, sensitivity, sulphur dioxide, tolerance     

Anther Culture of Lolium temulentum, Festuca pratensis and Lolium x Festuca Hybrids. I. Influence of Pretreatment, Culture Medium and Culture Incubation Conditions on Callus Production and Differentiation

This study was conducted with Lolium temulentum, Festuca pratensis,and the two hybrids L. multiflorum x F. pratensis ‘Elmet’and L. perenne x F. pratensis ‘Prior’. In a comparisonof various durations (7–42 d) of pretreatment at 4 or7 °C the highest yield of microspore-derived callus of L.temulentum was obtained after pretreatment of spikes at 7 °Cfor 28 d, conditions which also proved optimal for panicle pretreatmentwith F. pratensis. For ‘Elmet’, durations of 21–42d were optimal, and for ‘Prior’ the responses tendedto decline with increasing duration. In L. temulentum addition of charcoal (1–2 g l^–1)to medium containing 2, 4-D and KN wa     

Temperature Effects on Phenological Development and Yield of Muskmelon

Our goal was to construct a simple muskmelon phenology modelthat could be run with easily obtainable weather station dataand used by growers to quantify phenological development andaid in projecting harvest dates. A growth chamber experimentwas conducted with two cultivars of muskmelon (‘Gold Rush’and ‘Mission’) to determine how main vine leaf appearancerates responded to temperature. We identified three cardinaltemperatures for leaf appearance rate: the base temperature(10 °C) at which leaf appearance rate was zero; an optimumtemperature (34 °C) at which the rate of leaf appearancewas maximal; and an upper threshold temperature (45 °C)at which leaf appearance rate returned to zero. Using thesethree cardinal temperatures, we constructed a simplified thermalunit accumulator for hourly measurements of air temperature.Main vine plastochron interval (PI), thermal time to harvest,and final yield were determined for three cultivars of muskmelon(‘Explorer’, ‘Gold Rush’ and ‘Mission’)grown in the field at Overton, TX, USA, over six transplantingdates from March to June 1998. PI was calculated for each cultivarx transplanting date combination as the reciprocal of the slopeof main vine node number vs. accumulated hourly thermal units(     

Plant Morphology: The Historic Concepts of Wilhelm Troll, Walter Zimmermann and Agnes Arber

Recent molecular systematic and developmental genetic findingshave drawn attention to plant morphology as a discipline dealingwith the phenotypic appearance of plant forms. However, sincedifferent terms and conceptual frameworks have evolved overa period of more than 200 years, it is reasonable to surveythe history of plant morphology; this is the first of two paperswith this aim. The present paper deals with the historic conceptsof Troll, Zimmermann and Arber, which are based on Goethe'smorphology. Included are contrasting views of ‘unity anddiversity’, ‘position and process’, and ‘morphologyand phylogeny’, which, in part, are basic views of currentplant morphology, phylogenetic systematics and developmentalgenetics. Wilhelm Troll established the ‘type concept’and the ‘principle of variable proportions’. Hehas provided the most comprehensive overview of the positionalrelations of plant forms. Agnes Arber started from the universaldynamics of life and attempted to describe all structures asprocesses. She paid attention to ‘repetitive branching’,‘differential growth’, and ‘parallelism’.As a result she has recently been rediscovered by developmentalbotanists. Walter Zimmermann rejected any metaphysical influenceon plant form and instead called for objective procedures. Hewas mainly interested in phylogenetic ‘character transformation’and the ‘reconstruction of genealogical lines’.Guided by the example of flower-like inflorescences, a futurepaper will deal with functional and developmental constraintsinfluencing plant forms. Recent morphological concepts (‘trialectical’,‘continuum’/‘fuzzy’, ‘processmorphology’) will be discussed and related to currentmorphological and developmental genetic research. Copyright2001 Annals of Botany Company Plant form, plant morphology, natural philosophy, homology, phylogeny, Goethe, Troll, Arber, Zimmermann, typology, character transformation, differential growth, complementarity     

Increase in the Activity of Phenylalanine Ammonia-Lyase during the Non-Photoperiodic Induction of Flowering in Seedlings of Morning Glory (Pharbitis nil)

The activity of phenylalanine ammonia-lyase in the seedlingsof Pharbitis nil strains ‘Violet’ and ‘Kidachi’during the non-photoperiodic induction of flowering increasedin close association with the accumulation of phenylpropanoidsand the induction of flower buds. This observation suggestedthat the increased activity of phenyalanine ammonia-lyase causedthe accumulation of the phenylpropanoids and that the activationof the biosynthesis of phenylpropanoids might be involved inthe non-photoperiodic induction of flowering in the seedlingsof ‘Violet’ and ‘Kidachi’. (Received September 6, 1994; Accepted December 15, 1994)     

Diurnal Phototropism in Leaves of Lavatera cretica L. under Conditions of Simulated Solar-Tracking

Diurnal laminar reorientation was followed in solar-trackingleaves of Lavatera cretica L. under simulated conditions. Asimulated ‘sun’ was moved over the lamina in a 180?arc in the vertical plane of the mid-vein, at an angular velocityof 15? h^–1 in a regime of 12-h photoperiods. In one groupof plants the petioles of the experimental leaves were arrangedto face ‘sunrise’, while in the other they werearranged to face ‘sunset’. At ‘sunrise’,the laminae in both groups, which were inclined towards theanticipated direction of ‘sunrise’, changed theirelevation towards the rising ‘sun’, resulting inprogressive reduction in the angle of incidence (AI) of lighton the laminar surface (AI= differential between laminar and‘solar’ elevation). As a result, laminar and ‘solar’elevation converged, and laminar reorientation gradually ceased,until the ‘solar’ elevation had passed the normalto the laminar surface (AI=0?). laminar reorientation was thenresumed, but its direction was reversed to follow the directionof ‘solar’ reorientation. During most of the remaining‘day’, laminar elevation (LE) trailed that of the‘sun’ by an average of 11?-14?. Laminar reorientationthen anticipated ‘sunset’ by starting to slow down60 to 90min in advance. During the 12-h dark period, the laminareoriented towards the anticipated direction of the subsequent‘sunrise’. The time-course of nocturnal reorientationwas qualitatively different in the two groups of experimentalplants. The time-course of diurnal phototropism under naturaland simulated conditions is analysed and compared and differencesand similarities between them are discussed. Key words: Diurnal phototropism, solar-tracking, vectorial excitation     

ERRATA

WARBURG, M. R., 1965. On the water economy of some Australianland snails. Proc. malac. Soc. Lond. 36, 297–305. Page 298: second line from bottom, should read ‘within± 1 µg for Themapupa’. Page 300: Fig. 2 legend, should read ‘Evaporative waterloss from Sinumelon remissum (a), Pleuroxia sp. (b) and Themapupaadelaidae (c)’. Page 300: section 4 heading, should read ‘Continuous curvesfor water loss’. Page 301: second line, for ‘Fig. 9’ read ‘Fig.3’. Page 301: Table 1, last line, for ‘0.120024’ read‘0.12024’. Present address: Israel Institute for Biological Research, Ness-Ziona,Israel.     

Determinate Floral Buds of Plantain (Musa AAB) as a Site of Adventitious Shoot Formation

Floral buds of the ‘False Horn’ plantain clonesMusa (AAB) ‘Harton Verde’, ‘Harton Negra’,and ‘Currare’ terminate in a large single floralstructure. The apices of these floral buds are here designatedas determinate since they have lost the ability to produce additionalfloral initials or buds. Terminal peduncle segments can be culturedin a modified Murashige and Skoog (1962) medium supplementedwith N⁶-benzyl-aminopurine (5 mg I^–1). Under these conditions,this apparent inability to yield buds can be overcome as vegetativeshoot clusters form in the axils of the bracts. Rooted plantletsare obtainable by treating shoots with naphthaleneacetic acid(1 mg I^–1) and activated charcoal (0.025%). The adventitiousorigin of the shoots has been established. Musa cultivars, plantains, floral bud, adventitious buds, tissue culture     

Factors Affecting the Abscission of Reproductive Organs in Yellow Lupins (Lupinus luteus L.): III. ENDOGENOUS GROWTH SUBSTANCES IN VIRUS-INFECTED AND HEALTHY PLANTS AND THEIR EFFECT ON ABSCISSION

Extracts of small and mature-size lupin pods yielded four substancesaffecting the growth of wheat-coleoptile sections: one acidpromotor (A), two acid inhibitors(B and X), and one neutralinhibitor(Y). Inhibitor B was extremely active, however, coleoptile sectionsshowed no signs of toxic effects; they resumed growth at a rapidrate after rinsing them and adding ß-indolylaceticand (IAA) to the medium. 1 µg of IAA was required to counteractthe effect of ‘B’ extracted from 230 mg. Of tissue.On an equal fresh weight basis the inhibiting action of ‘B’in lupin pods was 500–1,500 times more potent than thatof ‘inhibitor ß’ in etiolated pea seedlings. Small pods of plants infected with pea-mosaic virus yielded3 times the amount of ‘A’ of healthy plants (equivalentto 1 µg. IAA 0.3 µg. IAA per 25 g. of tissue respectively),and approximately the amount of ‘B’. Mature podsof virus-infected plants again yielded more‘A’,but also 2? times more ‘B’ than pods of healthyplants. Healthy pods yielded more ‘A’ than virus-infectedpods, and there was no difference in ‘X’. A lupin abscission test was developed and the effects of proximaland distal application of -naphthyl acetic acid (NAA) are presented,and discussed with respect to results of other abscission tests. ‘A’ accelerated abscission when applied proximally,and delayed or prevented it when applied distally. ‘B’strongly accelerated abscission when applied in either way.A possible mechanism explaining the abscission-inducing effectof developing pods on later flowers is discussed in terms ofthe substances ‘A’ and ‘B’. The partlyprevented abscission observed on virus-infected plants was foundto agree well with the proposed mechanism.     

The Nitrogen Content of Plants and the Self-thinning Rule of Plant Ecology: A Test of the Core-skin Hypothesis

The ‘core-skin’ hypothesis postulates that secondarilythickened plants behave energetically as an inert ‘core’covered by an active ‘skin’, the ‘skin’being two-imensional, the ‘core’ three-dimensional.This would explain the ‘self-thinning ‘or‘–3/2’ rule of plant ecology, that is, the tendencyfor log (dry weight per plant) and log (number of plants perunit area) to progress along a straight line relationship, withslope = – 3/2’. The hypothesis was tested as follows. Plant nitrogen contentwas used as an estimate of the mass of ‘skin’ perplant, and dry weight as an estimate of the mass of the ‘core’.As plants mature the slope of the relationship between y = log(mass of nitrogen per plant) and x = log (mass of dry matterper plant) is expected to decline from an initial value of 1.0towards a final value of 0.66. The intercept of the relationshipis expected to reflect the intrinsic content of ‘skin’per unit of ‘core’. Genotypic variation in thisparameter should cause genotypic differences in the maximumattainable yield of biomass per unit area. The expectations were investigated by fitting the function y= p+qx+r exp – x to 30 sets of data on plant nitrogencontent, plant weight and time in 18 different vegetables. Simplelinear regressions of y on x were fitted to more limited setsof data on weights and nitrogen contents of mature trees. Theexpectations were, with some minor exceptions, confirmed. Nitrogen, yield, plant competition, self-thinning     

The Sensitivity of Net Photosynthesis in Several Plant Species to Short-term Fumigation with Sulphur Dioxide

The effects of exposure of up to 2 h with sulphur dioxide ona range of plant species was observed by measuring changes inthe rate of net photosynthesis under closely controlled environmentalconditions. Ryegrass, Lolium perenne ‘S23’ was thespecies most sensitive to SO₂; significant inhibition was detectedat 200 nl l^–1. Fumigations at 300 nl l^–1 also inhibitedphotosynthesis in field bean (Vicia faba cv. ‘Three FoldWhite’ and ‘Blaze’) and in barley (Hordeumvulgare cv. ‘Sonja’). No effect was detected inwheat (Triticum aestivum cv. ‘Virtue’) at concentrationsup to 600 nl l^–1 SO₂, or in oil-seed rape (Brassica napuscv. ‘Rafal’) except at 800 nl l^–1 SO₂). Recoverycommenced immediately after the fumigation was terminated andwas complete within 2 h when inhibition had not exceeded 20%during the SO₂ treatment. Key words: Sulphur dioxide, short-term fumigation, photosynthesis     

CORRECTIONS, VOLUME 29

Part 1, under the frontispiece portrait of Dr. N. B. Eales,the words ‘President 1948–1951’ should havebeen added. Page 103, line 49, for ‘Newton Collection’ read‘Norman Collection (Canon Norman)’. 185, line 37, for ‘capillaris’ read ‘capillacca’. 188, Table 1, for ‘bemoralis’. read ‘nemoralis’. 188, Table 2, for ‘Cochlicella acuta (Müll)? ventrosa(Fér.)’ read ‘Cochlicella ventrosa (Fér.)’. 191, line 24, for ‘araheo-’ read ‘archeo-’.     

Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Freezing tolerance, protein composition, and abscisic acid localization and content of pea epicotyl, shoot, and root tissue 3

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization