TheAfGene Regulates Timing and Direction of Major Developmental Events during Leaf Morphogenesis in Garden Pea (Pisum sativum)

The wildtype leaf of the garden pea possesses proximal pairsof leaflets and distal pairs of tendrils in the blade region.Theafila (af) mutation causes leaflets to be replaced by compound(branched) tendrils. We characterized the morphological variationin leaf form along the plant axis and leaf development in earlyand late postembryonic leaves onafilaplants to infer the roleof theAfgene. Leaf forms are more diverse early in shoot ontogenyonafilaplants.Afinfluences pinna length and pinna branchingin addition to pinna type. Pinna initiation in the proximalregion ofafilaleaf primordia is basipetal and delayed comparedto wildtype plants. In addition, pinna development in the proximalregion ofafilaleaves occurs for a longer period of time thanon wildtype leaf primordia. Therefore,Afregulates the timingand direction of leaf developmental processes in the proximalregion of the leaf, but has little effect on the distal region.These data support the heterochronic model of pea leaf morphogenesisproposed by Luet al. (International Journal of Plant Science157:311–355, 1996).Copyright 1999 Annals of Botany Company. afila,Fabaceae, garden pea, heterochrony, leaf morphogenesis,Pisum sativum.     

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     

Growth and Biomass Allocation, with Varying Nitrogen Availability, of Near-isogenic Pea Lines with Differing Foliage Structure

The single-gene mutation afila in pea (Pisum sativum L.) resultsin the replacement of proximal leaflets with branched tendrils,thereby reducing leaf area. This study investigated whethertheafila line could adjust biomass partitioning when exposedto varying nutrient regimes, to compensate for reduced leafarea, compared with wild-type plants. Wild-type and afila near-isogeniclines were grown in solution culture with nitrate-N added toinitially N-starved seedlings at relative addition rates (R_N)of 0.06, 0.12, 0.15 and 0.50 d^-1. The relative growth rate (R_W)of the whole plants closely matched R_Nat 0.06 and 0.12 d^-1,but higher R_Nresulted in a slightly higher growth rate. At agiven R_N, the wild-type line had lower plant nitrogen statusthan the afila line. R_Wof the roots of the afila line was lessthan R_Wof the roots of the wild-type at the three higher ratesof N supply despite a greater accumulation of N in the rootsof the afila plants. Consequently, plant nitrogen productivity(growth rate per unit nitrogen) was lower for afila. Dry matterallocation was strongly influenced by nitrogen status, but nodifferences in shoot–root dry matter allocation were foundbetween wild-type and afila with the same plant N status. Theseresults imply that decreased leaf area as a result of the single-genemutation afila affects dry matter allocation, but only accordingto its effect on the nitrogen status. Copyright 2000 Annalsof Botany Company Pisum sativum, pea, nitrogen limitation, growth, shoot–root allocation, relative growth rate, nitrogen productivity, isolines     

Leaf and Flower Development in Pea (Pisum sativum L.): Mutants cochleata andunifoliata

The stipule mutant cochleata(coch) and the simple-leaf mutantunifoliata(uni) are utilized to increase understanding of the controlof compound leaf and flower development in pea. The phenotypeof the coch mutant, which affects the basal stipules of thepea leaf, is described in detail. Mutant coch flowers have supernumeraryorgans, abnormal fusing of flower parts, mosaic organs and partialmale and female sterility. The wild-type Coch gene is shownto have a role in inflorescence development, floral organ identityand in the positioning of leaf parts. Changes in meristem sizemay be related to changes in leaf morphology. In the coch mutant,stipule primordia are small and their development is retardedin comparison with that of the first leaflet primordia. Thediameter of the shoot apical meristem of the uni mutant is approx.25% less than that of its wild-type siblings. This is the firsttime that a significant difference in apical meristem size hasbeen observed in a pea leaf mutant. Genetic controls in thebasal part of the leaf are illustrated by interactions betweencoch and other mutants. The mutantcoch gene is shown to changestipules into a more ‘compound leaf-like’ identitywhich is not affected by thestipules reduced mutation. The interactionof coch and tendril-less(tl) genes reveals that the expressionof the wild-type Tl gene is reduced at the base of the leaf,supporting the theories of gradients of gene action. Copyright2001 Annals of Botany Company Pisum sativum, garden pea, leaf morphogenesis, compound leaf, leaf mutants, flower morphology     

Roles of the Af and Tl Genes in Pea Leaf Morphogenesis: Shoot Ontogeny and Leaf Development in the Heterozygotes

The wildtype leaf blade of Pisum sativum possesses proximalleaflets and distal tendrils, which may be altered by two recessivemutations that affect pinna morphology, afila (afaf) and tendrilless(tltl). Using morphological observations and SEM, the variationin leaf forms along the plant axis and leaf development werecharacterized for plants heterozygous at the Af and/or Tl loci.The Af and Tl genes interacted to affect many characteristicsof shoot ontogeny, including rate changes in leaf blade lengthand complexity increases, as well as time to flowering. TheAf gene retarded early vegetative development and acceleratedthe time to flowering. The leaf phenotypes of these heterozygousgenotypes were specified mainly by changes in the timing ofmajor developmental events. The data support the hypothesesthat both genes are heterochronic in nature and that the pealeaf blade consists of three genetically- and developmentally-determined regions: proximal, distal and terminal. Copyright2000 Annals of Botany Company Heterochrony, leaf development, shoot ontogeny, Pisum sativum L., garden pea, afila,tendrilless .     

Pea Compound Leaf Architecture Is Regulated by Interactions among the Genes UNIFOLIATA, COCHLEATA, AFILA, and TENDRIL-LESS

The compound leaf primordium of pea represents a marginal blastozone that initiates organ primordia, in an acropetal manner, from its growing distal region. The UNIFOLIATA (UNI) gene is important in marginal blastozone maintenance because loss or reduction of its function results in uni mutant leaves of reduced complexity. In this study, we show that UNI is expressed in the leaf blastozone over the period in which organ primordia are initiated and is downregulated at the time of leaf primordium determination. Prolonged UNI expression was associated with increased blastozone activity in the complex leaves of afila (af), cochleata (coch), and afila tendril-less (af tl) mutant plants. Our analysis suggests that UNI expression is negatively regulated by COCH in stipule primordia, by AF in proximal leaflet primordia, and by AF and TL in distal and terminal tendril primordia. We propose that the control of UNI expression by AF, TL, and COCH is important in the regulation of blastozone activity and pattern formation in the compound leaf primordium of the pea.     

Differences in Architecture and Shoot Growth during Stagnant and Extension Growth Phases of Acanthopanax sciadophylloides(Araliaceae)

The shoot growth of a deciduous tree, Acanthopanax sciadophylloidesFranch. et Savat. shows inter-annual intermittent repetitionof two distinctive phases, a stagnant growth phase (S-phase)and vigorous extension-growth (E-phase). To help understandthe differentiation mechanism, shoot development was studiedover time in both shoot phases. S-phase and E-phase shoots weredistinguished from each other by morphological traits: S-phaseshoots are characterized by higher allocation to leaves anda shorter period of stem growth, while E-phase shoots show continuousstem extension over the growing season. Specific leaf area didnot differ between the two phases. This shoot differentiationwas similar to the morphological differentiation of shoots betweenlong vs. short shoots found in some temperate trees. Leavesof both phases were well-dispersed through adjustment of petiolelength and leaf-blade size to reduce mutual shading within ashoot. Stem-wood density of current-year shoots was lower inE-phase compared with S-phase shoots. Leaves produced earlyin the season affected the growth phase of the following year.These results suggest that annual shoot differentiation of A.sciadophylloides was determined during the previous season andreflects leaf productivity in a given habitat during that growingseason. Copyright 2001 Annals of Botany Company Acanthopanax sciadophylloides, Araliaceae, biomass allocation, intermittent shoot growth, leaf display, shoot architecture, shoot differentiation     

The rates of shoot and root growth in intact plants of pea mutants in leaf morphology

Isogenic lines of pea (Pisum sativum L.) with the genetically determined changes in leaf morphology, afila (af) and tendril-less (tl), were used to study the relationship between shoot and root growth rates. The time-course of shoot and root growth was followed during the pre-floral period in the intact plants grown under similar conditions. The af mutation produced afila leaves without leaflets, whereas in the case of the tl mutations, tendrils were substituted with leaflets, and acacia-like leaves were developed. Due to the changes in leaf morphology caused by these mutations, pea genotypes differed in leaf area: starting from day 7, the leaf area was lower in the af plants and larger in the tl plants as compared to the wild-type plants. Such divergence was amplified in the course of plant development and reached its maximum immediately before the transition to flowering. Plants of isogenic lines did not notably differ in stem surface areas. In spite of significant difference in total leaf area, the wild type and tl plants did not differ in leaf dry weight. Starting from leaf 9, the af plants lagged behind two leaflet-bearing genotypes (wild type and tl) in leaf dry weight, whereas stem dry weight was similar in the wild type and tl forms and slightly lower in the af plants. Root dry weights were practically similar in the wild type and tl plants until flowering. The reduction of leaf area in the af plants drastically reduced root dry weight. In other words, the latter index was related to the total weight and total area of leaves and stems. The correlation analysis demonstrated an extremely low relationship between leaf and stem area and dry weight and those of roots early in plant development (when plants develop five to seven leaves). Later, immediately before flowering (nine to eleven leaves), root weight was positively related to leaf weight and area; however, stem area and root weight did not correlate. Thus, in three genotypes (wild type, af, and tl), at the end of their vegetative growth phase, leaf and root biomass accumulated in proportion, independently of leaf area expansion.     

An Analysis of Spatial Variation in the Nitrogen Content of Leaves from Different Horizons Within a Canopy

An analysis of spatial variation in the specific nitrogen contentof leaves from different positions within a canopy is developed.It is used to examine data obtained for contrasting crop species;the legume crop guar (Cyamopsis tetragonoloba), and crops ofa forage sorghum. The analysis distinguishes two componentsof the leaf nitrogen content: one associated with the metabolicapparatus within the leaf tissues and the other with structuralelements of the leaf. The analysis allows the spatial variationin specific leaf nitrogen content to be quantitatively described. Leaf nitrogen content, crop growth, light interception     

Sunflower (Helianthus annuus) Leaves Contain Compounds that Reduce Nuclear Propidium Iodide Fluorescence

Sunflower leaves have unidentified compounds that interferewith propidium iodide (PI) intercalation and/or fluorescence.Independently prepared pea leaf nuclei show greater PI fluorescencethan nuclei from pea leaves simultaneously processed (co-chopped)with sunflower leaves. Differences in fluorescence persist aftermixing the PI-stained pea and the co-chopped pea/sunflower samples,i.e. PI staining protects the nuclei from the effects of theinhibitor. The current results are significant to practicalflow cytometric determination of plant nuclear DNA content.They show: (1) simultaneous processing of nuclear samples fromthe target and the standard species is necessary to obtain reliableDNA estimates; (2) a test for the presence of inhibitors shouldbe conducted; and (3) when inhibitors are present caution shouldbe taken in interpreting differences in estimated DNA content.The previously reported environmentally-induced variation inDNA content in sunflower populations is most simply explainedby variation in the amount of environmentally-induced inhibitorthat interferes with intercalation and/or fluorescence of PI.Intraspecific variation of DNA content for Helianthus annuusneeds to be re-evaluated using best practice techniques comparingphysiologically uniform tissues that are free of inhibitors.The best estimate for 2C DNA content of H. annuus used in thisstudy is 7.3 pg. Copyright 2000 Annals of Botany Company Helianthus annuus, DNA content, flow cytometry, propidium iodide, endogenous inhibitors     

Effects of Gibberellin on Shoot Development in the dgt Mutant of Tomato

The morphological effects of gibberellin A₃ (GA₃) on the dgtmutant of tomato were investigated. The mutant effectively showedthe normal range of responses, including a promotion of stemlength due to an increased number of longer internodes, a dramaticincrease in apical dominance, and effects on leaf shape andcolour. In the case of stem elongation, the quantitative responseof the mutant was greater than normal. The morphological abnormalitiescharacteristic of the dgt mutant, such as horizontal growth,a thin stem and hyponastic leaves, were not normalized by GA₃. It is concluded that the demonstrated lack of response to auxinof the dgt mutant does not impair its gibberellin responses. Tomato, gibberellin, auxin, mutant, shoot development     

Modification of Leaf Formation by Cytokinin and Chlormequat (CCC) in Vitis

Application of the cytokinin, 6-(benzylamino)-9-(2-tetrahydropyrany1)-9H-purine(PBA, 1mM) and 2-chloroethyl trimethyl ammonium chloride (chlormequat;3 mM), a growth retardant, to grapevines induced the formationof fused leaves (two laminae and two petioles) and double leaves(two separate petioles each with a single lamina). Double leaveswere found in Vitis vinifera L. seedlings, in Vitis ripariaand in Muscadinia (Vitis) rotundifolia. In some of the treatedvines, leaves arose with opposite phyllotaxy. Other anomaliesincluded production of two opposite axillary buds in axils ofopposite leaves and production of two tendrils per node. Sometendrils grew into shoots. In Muscat of Alexandria, applicationof GA₃ (3 µM, 15 µM), followed by an applicationof chlormequat (0.4 mM, 1.2 mM), led to a marked reduction inthe extension growth of axillary shoots. Vitis vinifera L., Vitis riparia, Muscadinia (Vitis) rotundifolia, grapevines, leaf formation, cytokinin, chloromequat     

Does Cytokinin Transport from Root-To-Shoot in the Xylem Sap Regulate Leaf Responses to Root Hypoxia?

We have examined the hypothesis that cytokinins transportedfrom roots to shoots affects leaf growth, stomatal conductance,and cytokinin concentration of leaves of Phaseolus and a hybridpoplar (Populus trichocarpa x Populus deltoides) with hypoxicroots. Because cytokinins may interact with other substances,potassium and calcium concentrations were determined in xylemsap of Populus plants with hypoxic and aerated roots while gibberellin(GA) concentrations were measured in shoot tissues. Root hypoxiadecreased leaf growth and closed stomata in both species. Inboth species, fluxes of cytokinins out of the roots were reduced,but no differences in bulk leaf concentrations were measuredbetween the hypoxic and aerated plants. Shoots with aeratedroots contained slightly higher concentrations of GA₁ and GA₃than shoots from hypoxic plants. There were no differences incalcium or potassium concentrations in xylem sap between aerationtreatments. Exogenously applied cytokinins did not alleviatethe growth or stomatal responses caused by root hypoxia. Informationon the site(s) and mechanism(s) of cytokinin action and theways in which cytokinins are compartmentalized within plantcells will be required to understand the physiological significanceof cytokinin transport in the transpirational stream. Key words: Cytokinins, hypoxia, Populus, Phaseolus     

The Effect of Light Quality on Shoot Extension Growth in Three Species of Grasses

The effect of light quality on the extension growth of vegetativeshoots and on the final size of their leaves was investigatedin plants of Lolium multiflorum, Sporobolus indicus and Paspalumdilatatum. Three experimental approaches were used, (a) redor far-red end-of-day irradiations of sunlight-grown plants,(b) different red/far-red ratios of white light in a growthroom and (c) sunlight enrichment with radiation of differentred/far-red ratios or with different amounts of far-red lightduring the photoperiod. Plants treated with end-of-day far-redor low red/far-red ratios throughout the photoperiod developedlonger leaves and, as a result, longer shoots. This effect wasmore marked in leaf sheaths than in blades. Tiller extensionand leaf sheath length increased with the amount of far-redadded to sunlight in a simple hyperbolic relationship. Theseresults show that vegetative grass shoots respond to light qualityin a way similar to internodes of dicotyledonous plants. Lolium multiflorum Lam., Sporobolus indicus (L.), Paspalum dilatatum (Poir.), leaf growth, tiller growth, photomorphogenesis     

Identification of a Novel Triterpenoid Saponin from Pisum sativum as a Specific Inhibitor of the Diguanylate Cyclase of Acetobacter xylinum

A specific and highly potent inhibitor of diguanylate cyclase,the key regulatory enzyme of the cellulose synthesizing apparatusin the bacterium Acetobacter xylinum, was isolated from extractsof etiolated pea shoots (Pisum sativum). The inhibitor has beenpurified by a multistep procedure, and sufficient amounts ofhighly purified compound (3-8 mg) for spectral analysis wereobtained. The structure of this compound was established as3-O-a-L-rhamnopyranosyl-(l     

Leaf Production and Morphology in the Artichoke Gall of Yew (Taxus baccata L.)

Comparative leaf production rates and leaf morphology studiesfor galled and normal shoots of yew trees have been obtainedthroughout the life cycle of the causative agent, the gall midge,Taxomyia taxi. Normal and galled shoot leaf numbers have beenrelated to those of their parent shoots. It was found that whereasthe annual leaf production of normal shoots was positively relatedto that of the parent shoots, galled shoot leaf production remainedconstant regardless of parent shoot vigour showing leaf stimulationby the midge to be a special case. The midge larva appears tobe determining the rate of leaf production in galled shoots.In galls leaf production continues throughout the winter monthswith no dormant period. From morphological evidence, alternationof leaves and cataphylls is continued in galled buds.     

Can early wilting of old leaves account for much of the ABA accumulation in flooded pea plants?

When pea plants (Pisum sativum L.) were subjected to flooding,abscisic acid (ABA) content in shoots and roots increased upto 8-fold in the following days and stomatal conductance significantlydecreased. Although young leaves of flooded plants had a slightlyhigher water potential than those of the unflooded plants, oldleaves had lower water potential and lost turgor at the timewhen a substantial ABA increase was detected. In plants wherethe old leaves were clipped off, flooding did not cause anyABA increase during 7 d of the experimental period, except underconditions of higher transpiration demand, when the increasein ABA content was both delayed and small in scale (only I-fold).When intact plants were flooded and ABA was assayed separatelyin both old and young leaves, the ABA increase in old leavespreceded that in young leaves. Evidence here suggests that theflooding-induced ABA increase mainly results from the wiltingof old leaves. This suggests that young leaves may be protectedfrom wilting by ABA originating in old leaves under unfavourableenvironmental conditions. Key words: Waterlogging, soil flooding, ABA, leaf water relations, pea, Pisum sativum     

Vegetative Anatomy of Uncinia (Cyperaceae)

The vegetative anatomy of 18 (19) Uncinia spp. (Cyperaceae-Cariceae)including representatives of most taxonomic subdivisions wasstudied to determine the range of variation in certain anatomicaland morphological characters of the vegetative organs withinthe genus. The two South American species U. erinacea and U. kingii differfrom all others, the former in having a closed cylinder of sclerenchymain the culm, and the latter a grooved culm. The three tall SouthAmerican spp., U. brevicaulis, U. hamata, and U. phleoides var.trichocarpa, are characterized by adaxial intercostal fibrestrands in the leaves, and in this respect show affinities withtall New Zealand species, U. sinclairii and U. uncinata. Thelast species does not have the intercostal strands. The muchsmaller U. tenuis from S. America resembles, in the size andshape of transverse sections of leaf and culm, a group of speciesfrom New Zealand comprising U. angustifolia, U. egmontiana,and U. rupestris. To these could be added U. banksii, U. hookeri,and perhaps also U. tenella, although the last two spp. exhibitsome distinctive characters in the transverse section of theleaf and also in the leaf epidermis in surface view. No exactcounterpart to the Australian and New Zealand spp. U. divaricata,U. riparia, U. rubra and U. scabra was found amongst the S.American material. This group of spp. is distinctive, for example,because of the triangular or irregularly triangular shape oftransverse sections of the culm and the large amount of sclerenchymain transverse sections of the leaf. The range of structural variation appears to be particularlywide in the S. American spp., which represent two extreme typesin the shape of the leaf in transverse section. The other talland small species have their respective counterparts in eacharea, although they are more profusely represented in New Zealandthan in America, and therefore show a correspondingly greaterrange of structure.