Vegetative Meristem Research Articles

Characterization of aFLORICAULA/LEAFYHomologue ofGnetum parvifoliumand Its Implications for the Evolution of Reproductive Organs in Seed Plants

The morphological variation among reproductive organs of extant seed plants makes assessment of organ homology difficult. Comparisons of expression patterns of homeotic genes that control organ development will yield new information about the homology of organs to assess inferences deduced from previous morphological studies. In angiosperms, the FLORICAULA/LEAFY (FLO/LFY) genes convert a vegetative shoot meristem to a floral meristem by inducing floral homeotic genes, most of which belong to the MADS‐box gene family. To provide insights into the evolution of reproductive organs in seed plants, a FLO/LFY homologue (GpLFY) was cloned from Gnetum parvifolium. GpLFY mRNA was expressed in both the vegetative shoot apex and the female strobilus. The GpLFY mRNA signal was detected in early developmental stages of the collar and the ovule primordium, including the nucellus and three envelopes. A comparison of FLO/LFY gene expression in Gnetum and the conifer Pinus radiata indicates that the Gnetum collar and ovul...

KNAT2

The KNAT2 (for KNOTTED-like from Arabidopsis thaliana 2) homeobox gene is expressed in the vegetative apical meristem. It is also active during flower development, suggesting a function in the structuring of flowers. To investigate its role, we used a DEXAMETHASONE (DEX)-inducible system to generate transgenic plants that overexpressed a fusion of KNAT2 with the hormone binding domain of the glucocorticoid receptor. DEX-induced plants were similar to plants overexpressing the closely related KNAT1 gene, indicating overlapping functions, although we observed differences as well. In particular, KNAT2-GR activation induced ectopic carpel features. First, KNAT2 induced the homeotic conversion of nucellus into carpel-like structures. Second, KNAT2 induced stigmatic papillae on rosette leaves in the ap2-5 background. Third, ectopic expression of the carpel identity gene AGAMOUS (AG) was observed in carpels and ovules. Interestingly, the homeotic conversion was not dependent on AG activity, because it was maintained in the ag-1 ap2-5 double mutant. Therefore, our data indicate that KNAT2 also must activate other carpel regulators. Together, these results suggest that KNAT2 plays a role in carpel development.

Evolution of Floral Meristem Identity Genes. Analysis ofLolium temulentum Genes Related to APETALA1 andLEAFY of Arabidopsis

Flowering (inflorescence formation) of the grass Lolium temulentum is strictly regulated, occurring rapidly on exposure to a single long day (LD). During floral induction, L. temulentum differs significantly from dicot species such as Arabidopsis in the expression, at the shoot apex, of two APETALA1 (AP1)-like genes, LtMADS1 and LtMADS2, and of L. temulentum LEAFY (LtLFY). As shown by in situ hybridization, LtMADS1 and LtMADS2 are expressed in the vegetative shoot apical meristem, but expression increases strongly within 30 h of LD floral induction. Later in floral development, LtMADS1 and LtMADS2 are expressed within spikelet and floret meristems and in the glume and lemma primordia. It is interesting that LtLFY is detected quite late (about 12 d after LD induction) within the spikelet meristems, glumes, and lemma primordia. These patterns contrast with Arabidopsis, where LFY and AP1 are consecutively activated early during flower formation. LtMADS2, when expressed in transgenic Arabidopsis plants under the control of the AP1 promoter, could partially complement the organ number defect of the severe ap1-15 mutant allele, confirming a close relationship between LtMADS2 and AP1.

WAKs: cell wall-associated kinases linking the cytoplasm to the extracellular matrix.

There are only a few proteins identified at the cell surface that could directly regulate plant cell wall functions. The cell wall-associated kinases (WAKs) of angiosperms physically link the plasma membrane to the carbohydrate matrix and are unique in that they have the potential to directly signal cellular events through their cytoplasmic kinase domain. In Arabidopsis there are five WAKs and each has a cytoplasmic serine/threonine protein kinase domain, spans the plasma membrane, and extends a domain into the cell wall. The WAK extracellular domain is variable among the five isoforms, and collectively the family is expressed in most vegetative tissues. WAK1 and WAK2 are the most ubiquitously and abundantly expressed of the five tandemly arrayed genes, and their messages are present in vegetative meristems, junctions of organ types, and areas of cell expansion. They are also induced by pathogen infection and wounding. Recent experiments demonstrate that antisense WAK expression leads to a reduction in WAK protein levels and the loss of cell expansion. A large amount of WAK is covalently linked to pectin, and most WAK that is bound to pectin is also phosphorylated. In addition, one WAK isoform binds to a secreted glycine-rich protein (GRP). The data support a model where WAK is bound to GRP as a phosphorylated kinase, and also binds to pectin. How WAKs are involved in signaling from the pectin extracellular matrix in coordination with GRPs will be key to our understanding of the cell wall's role in cell growth.

Florogenesis and the effect of temperatures on the development of Allium aflatunense

SummaryThe process of florogenesis of Allium aflatunense (= A. hollandicum) and the effects of temperature and photoperiod on plant development were studied., Intrabulb development of the monocarpic shoot, and the formation of the renewal bulbs of A. aflatunense require a thermoperiodic annual cycle. Initiation of leaf primordia in the renewal bulb begins after planting of the parent plant. The vegetative meristem of the renewal bulb shifts to the reproductive stage during flowering of the parent plant, while the differentiation of individual flowers in the floral meristem begins during the rest period of the bulb, when temperatures are relatively high. New flower primordia are continuously being formed, while older flower buds have already been diferentiated. After floral differentiation, a prolonged period at low temperatures is required for further leaf and floral stalk elongation. After planting, the physiological response of the growing plants was affected by the temperature and length of preceding storage. Storage at 4˚C for 16 weeks, or a combination of 98C for eight weeks, followed by 4˚C for eight weeks, resulted in normal leaf and floral stalk elongation. Leaf elongation requires shorter cold treatment than that of the floral stalk. Storage conditions conducive to scape elongation also resulted in the formation of a renewal bulb and a few daughter bulbs. The development of the leaves, renewal bulbs and inflorescences was significantly affected by growth temperatures, but not by photoperiod. Leaf and renewal bulb growth was best at 17/98C (day/night temperatures), intermediate at 20/12˚C and 23/15˚C, and weakest at 26/18˚C, where leaves were narrower and shorter and renewal bulbs were smaller than in the other environments. Scape elongation was normal at 17/9˚C and 20/12˚C, and inhibited in plants subjected to higher temperatures.

Sexual development and sex chromosomes in hop.

The stages of floral development in staminate and pistillate plants of hop (Humulus lupulus) were defined using scanning electron microscopy and light microscopy. Vegetative meristems of male and female plants are morphologically indistinguishable. On transition to the reproductive phase, inflorescence apices reduce greatly in size and striking developmental sex differences become apparent. The first sex-specific differences occur extremely early in floral ontogeny. Both male and female plants initiate inflorescence meristems at each leaf node, each meristem being enclosed within a bract. Male secondary inflorescence meristems give rise to clusters of asynchronously developing flowers. Female inflorescence meristems produce flowers arranged in 'cones'. Each male floral meristem initiates a whorl of five sepal primordia, followed by an inner whorl of five stamen primordia. There is no sign of carpel development at any stage. In females, two carpel primordia are initiated, surrounded at their base by a vestigial perianth whorl. No stamen development is observed. Several monoecious lines carry bisexual flowers, either within cymose panicles or within the basal bracts of terminal female inflorescences. Bisexual flowers usually possess perianth, stamen and carpel whorls. The central whorls are often highly variable, and range from a pair of stigmas fused to a thin central filament to a well developed gynoecium. Chimaeric central whorls consisting of fused staminoid-carpelloid structures also occur. Sex differences in unisexual hop flowers are determined at an extremely early stage in ontogeny. The inappropriate set of sex organs is suppressed before it becomes visible or, more probably, it is not initiated at all. Genes directing the development of sex are likely to act at an extremely early stage, well in advance of floral organogenesis. The sex chromosomes of dioecious hop plants are described, as well as the chromosome constitutions of monoecious plants and those carrying bisexual flowers.

Differential gene expression during floral transition in an orchid hybrid Dendrobium Madame Thong-In.

We have developed an in vitro orchid (Dendrobium grex Madame Thong-In) system as a model for investigating gene expression during floral transition. Total RNA was isolated from vegetative shoot apical meristems (VSAM) and transitional shoot apical meristems (TSAM). Using the mRNA differential display method, we identified 53 cDNA clones differentially expressed in VSAMs and 16 cDNA fragments specifically expressed in TSAMs. Northern blot analysis confirmed that, on the basis of their transcripts, 12 cDNAs decreased and 8 cDNAs increased during floral transition. Sequence analysis suggested that 5 clones are likely to be transcription factors, including one MADS-box gene of the AGL2 subfamily, one class 1 knox gene and one homolog of the Drosophila seven-up gene. Other clones showed homology with those coding for Arabidopsis SOD copper chaperone, CKI and the tobacco 21D7 protein.

ACAULIS5, an Arabidopsis gene required for stem elongation, encodes a spermine synthase.

Polyamines have been implicated in a wide range of biological processes, including growth and development in bacteria and animals, but their function in higher plants is unclear. Here we show that the Arabidopsis: ACAULIS5 (ACL5) gene, whose inactivation causes a defect in the elongation of stem internodes by reducing cell expansion, encodes a protein that shares sequence similarity with the polyamine biosynthetic enzymes spermidine synthase and spermine synthase. Expression of the recombinant ACL5 protein in Escherichia coli showed that ACL5 possesses spermine synthase activity. Restoration of the acl5 mutant phenotype by somatic reversion of a transposon-induced allele suggests a non-cell-autonomous function for the ACL5 gene product. We also found that expression of the ACL5 cDNA under the control of a heat shock gene promoter in acl5 mutant plants restores the phenotype in a heat shock-dependent manner. The results of the experiments showed that polyamines play an essential role in promotion of internode elongation through cell expansion in Arabidopsis: We discuss the relationships to plant growth regulators such as auxin and gibberellins that have related functions.

Identification and Characterization of Three Orchid MADS-Box Genes of the AP1/AGL9 Subfamily during Floral Transition

Gene expressions associated with in vitro floral transition in an orchid hybrid (Dendrobium grex Madame Thong-In) were investigated by differential display. One clone, orchid transitional growth related gene 7 (otg7), encoding a new MADS-box gene, was identified to be specifically expressed in the transitional shoot apical meristem (TSAM). Using this clone as a probe, three orchid MADS-box genes, DOMADS1, DOMADS2, and DOMADS3, were subsequently isolated from the TSAM cDNA library. Phylogenetic analyses show that DOMADS1 and DOMADS2 are new members of the AGL2 subfamily and SQUA subfamily, respectively. DOMADS3 contains the signature amino acids as with the members in the independent OSMADS1 subfamily separated from the AGL2 subfamily. All three of the DOMADS genes were expressed in the TSAM during floral transition and later in mature flowers. DOMADS1 RNA was uniformly expressed in both of the inflorescence meristem and the floral primordium and later localized in all of the floral organs. DOMADS2 showed a novel expression pattern that has not been previously characterized for any other MADS-box genes. DOMADS2 transcript was expressed early in the 6-week-old vegetative shoot apical meristem in which the obvious morphological change to floral development had yet to occur. It was expressed throughout the process of floral transition and later in the columns of mature flowers. The onset of DOMADS3 transcription was in the early TSAM at the stage before the differentiation of the first flower primordium. Later, DOMADS3 transcript was only detectable in the pedicel tissues. Our results suggest that the DOMADS genes play important roles in the process of floral transition.

Auxin regulates the initiation and radial position of plant lateral organs.

Leaves originate from the shoot apical meristem, a small mound of undifferentiated tissue at the tip of the stem. Leaf formation begins with the selection of a group of founder cells in the so-called peripheral zone at the flank of the meristem, followed by the initiation of local growth and finally morphogenesis of the resulting bulge into a differentiated leaf. Whereas the mechanisms controlling the switch between meristem propagation and leaf initiation are being identified by genetic and molecular analyses, the radial positioning of leaves, known as phyllotaxis, remains poorly understood. Hormones, especially auxin and gibberellin, are known to influence phyllotaxis, but their specific role in the determination of organ position is not clear. We show that inhibition of polar auxin transport blocks leaf formation at the vegetative tomato meristem, resulting in pinlike naked stems with an intact meristem at the tip. Microapplication of the natural auxin indole-3-acetic acid (IAA) to the apex of such pins restores leaf formation. Similarly, exogenous IAA induces flower formation on Arabidopsis pin-formed1-1 inflorescence apices, which are blocked in flower formation because of a mutation in a putative auxin transport protein. Our results show that auxin is required for and sufficient to induce organogenesis both in the vegetative tomato meristem and in the Arabidopsis inflorescence meristem. In this study, organogenesis always strictly coincided with the site of IAA application in the radial dimension, whereas in the apical-basal dimension, organ formation always occurred at a fixed distance from the summit of the meristem. We propose that auxin determines the radial position and the size of lateral organs but not the apical-basal position or the identity of the induced structures.

Herbivory could unlock mutations sequestered in stratified shoot apices of genetic mosaics

Many higher plants have shoot apical meristems that possess discrete cell layers, only one of which normally gives rise to gametes following the transition from vegetative meristem to floral meristem. Consequently, when mutations occur in the meristems of sexually reproducing plants, they may or may not have an evolutionary impact, depending on the apical layer in which they reside. In order to determine whether developmentally sequestered mutations could be released by herbivory (i.e., meristem destruction), a characterized genetic mosaic was subjected to simulated herbivory. Many plants develop two shoot meristems in the leaf axils of some nodes, here referred to as the primary and secondary axillary meristems. Destruction of the terminal and primary axillary meristems led to the outgrowth of secondary axillary meristems. Seed derived from secondary axillary meristems was not always descended from the second apical cell layer of the terminal shoot meristem as is expected for terminal and primary shoot meristems. Vegetative and reproductive analysis indicated that secondary meristems did not maintain the same order of cell layers present in the terminal shoot meristem. In secondary meristems reproductively sequestered cell layers possessing mutant cells can be repositioned into gamete-forming cell layers, thereby adding mutant genes into the gene pool. Herbivores feeding on shoot tips may influence plant evolution by causing the outgrowth of secondary axillary meristems.

Isolation and Characterization of a MADS-box Gene cDNA, PnMADS1, That is Expressed in Both Vegetative and Floral Meristems of Pharbitis nil.

A novel cDNA corresponding to a gene designated PnMADS1 and encoding a MADS-box protein was isolated from the short-day plant Pharbitis nil (Japanese morning glory) cv. Violet. Phylogenetic analysis of the amino acid sequence encoded by PnMADS1 indicated that PnMADS1 does not belong to any of the major sub-groups of MADS-box proteins. The protein exhibited some similarity to proteins in the so-called ‘orphan group’ of unclassified MADS-box proteins that are mainly expressed in vegetative organs. The most similar MADS-box genes were AGL24 and StMADS16 (53.2% and 52.3% identity at the deduced amino-acid level, respectively). We obtained evidence that PnMADS1 mRNA accumulates in both vegetative and floral meristems but not in any other vegetative organs. Our results indicate that PnMADS1 is a novel MADS-box protein in terms of both structure and pattern of expression.

Floral Ontogeny of Pelargonium ×domesticum

Floral ontogeny of two cultivars of Pelargonium ×domesticum L.H. Bailey, (regal pelargonium) `Duchess' and `Jennifer', was examined. Plants of both cultivars were grown together in a growth chamber at 15.5 °C with a photosynthetic photon flux of 10 mol·m-2·d-1. Meristems were examined at 5-day intervals over an experimental period of 170 days. The initial vegetative meristem was convex with leaf primordia initiated on either side in an alternate pattern. Early floral initiation was characterized by formation of two clefts on either side of the meristem. Between the clefts new meristems developed. Proliferation of meristems continued until numerous meristems were organized in a cluster arrangement at the apex of the shoot. New meristems lacked leaf primordia and would develop into flowers. Floral organ primordia on a floral meristem were initiated in a succession of four whorls: sepals, petals, androecia, and gynoecium.

Disturbance in the allocation of carbohydrates to regenerative organs in transgenic Nicotiana tabacum L.

Transgenic tobacco (Nicotiana tabacum L, cv. SR-1) expressing mannitol 1-phosphate dehydrogenase, MTLD, in chloroplasts and myo-inositol O-methyltransferase, IMT1, in the cytosol after crossing of lines which expressed these foreign genes separately has been analysed. Plants expressing both enzymes accumulated mannitol and D-ononitol in amounts comparable to those following single gene transfer and showed phenotypically normal growth during the vegetative stage. Induction of flowering for transgenovar and wild-type occurred at the same time, but during flowering the phenotype of the transformed plants changed. Compared to wild-type, transgenic plants were characterized by curled, smaller upper leaves and elongated stems during flowering; incomplete development of flower buds with shorter sepals and pedicels resulted in increased abortion. Flowers completing development were normal. The vegetative biomass of the transformed plants was slightly higher than that of wild-type. Concentrations of soluble sugars and potassium were lower than in wild-type only in the apical parts of the transgenic plants. Both enzymes, under control of the CaMV 35S promoter, promoted accumulation of mannitol and D-ononitol in the youngest leaves close to the vegetative meristem and in flowers, suggesting that their presence could signal lower sink demand leading to a decrease in carbon import to flowers and developing seed capsules. The interpretation here is that increases of inert carbohydrates in developing sinks interfere with metabolism, such as respiration or glycolysis. This interference may be less significant in source tissues during vegetative growth than in sink tissues during seed development.

FALSIFLORA, the tomato orthologue of FLORICAULA and LEAFY, controls flowering time and floral meristem identity.

Characterization of the tomato falsiflora mutant shows that fa mutation mainly alters the development of the inflorescence resulting in the replacement of flowers by secondary shoots, but also produces a late-flowering phenotype with an increased number of leaves below first and successive inflorescences. This pattern suggests that the FALSIFLORA (FA) locus regulates both floral meristem identity and flowering time in tomato in a similar way to the floral identity genes FLORICAULA (FLO) of Antirrhinum and LEAFY (LFY) of Arabidopsis. To analyse whether the fa phenotype is the result of a mutation in the tomato FLO/LFY gene, we have cloned and analysed the tomato FLO/LFY homologue (TOFL) in both wild-type and fa plants following a candidate gene strategy. The wild-type gene is predicted to encode a protein sharing 90% identity with NFL1 and ALF, the FLO/LFY-like proteins in Nicotiana and Petunia, and about 80 and 70% identity with either FLO or LFY. In the fa mutant, however, the gene showed a 16 bp deletion that results in a frameshift mutation and in a truncated protein. The co-segregation of this deletion with the fa phenotype in a total of 240 F2 plants analysed supports the idea that FA is the tomato orthologue to FLO and LFY. The gene is expressed in both vegetative and floral meristems, in leaf primordia and leaves, and in the four floral organs. The function of this gene in comparison with other FLO/LFY orthologues is analysed in tomato, a plant with a sympodial growth habit and a cymose inflorescence development.

A petunia MADS box gene involved in the transition from vegetative to reproductive development.

We have identified a novel petunia MADS box gene, PETUNIA FLOWERING GENE (PFG), which is involved in the transition from vegetative to reproductive development. PFG is expressed in the entire plant except stamens, roots and seedlings. Highest expression levels of PFG are found in vegetative and inflorescence meristems. Inhibition of PFG expression in transgenic plants, using a cosuppression strategy, resulted in a unique nonflowering phenotype. Homozygous pfg cosuppression plants are blocked in the formation of inflorescences and maintain vegetative growth. In these mutants, the expression of both PFG and the MADS box gene FLORAL BINDING PROTEIN26 (FBP26), the putative petunia homolog of SQUAMOSA from Antirrhinum, are down-regulated. In hemizygous pfg cosuppression plants initially a few flowers are formed, after which the meristem reverts to the vegetative phase. This reverted phenotype suggests that PFG, besides being required for floral transition, is also required to maintain the reproductive identity after this transition. The position of PFG in the hierarchy of genes controlling floral meristem development was investigated using a double mutant of the floral meristem identity mutant aberrant leaf and flower (alf) and the pfg cosuppression mutant. This analysis revealed that the pfg cosuppression phenotype is epistatic to the alf mutant phenotype, indicating that PFG acts early in the transition to flowering. These results suggest that the petunia MADS box gene, PFG, functions as an inflorescence meristem identity gene required for the transition of the vegetative shoot apex to the reproductive phase and the maintenance of reproductive identity.

CYP78A5 encodes a cytochrome P450 that marks the shoot apical meristem boundary in Arabidopsis.

The normal development of shoot structures depends on controlling the growth, proliferation and differentiation of cells derived from the shoot apical meristem. We have identified the CYP78A5 gene encoding a putative cytochrome P450 monooxygenase that is the first member of the CYP78 family from Arabidopsis. This gene is strongly expressed in the peripheral regions of the vegetative and reproductive shoot apical meristems, defining a boundary between the central meristematic zone and the developing organ primordia. In addition, CYP78A5 shows a dynamic pattern of expression during floral development. Overexpression of CYP78A5 affects multiple cell types, causing twisting and kinking of the stem and defects in floral development. To define the relationship of CYP78A5 to genes controlling meristem function, we examined CYP78A5 expression in plants mutant for SHOOT MERISTEMLESS, ZWILLE and ARGONAUTE, and have found that CYP78A5 expression is altered in these mutant backgrounds. We propose that CYP78A5 has a role in regulating directional growth in the peripheral region of the shoot apical meristem in response to cues established by genes regulating meristem function.

Evolution of phenotypic plasticity a comparative approach in the phylogenetic neighbourhood of Arabidopsis thaliana

Abstract The evolution of phenotypic plasticity has rarely been examined within an explicitly phylogenetic framework, making use of modern comparative techniques. Therefore, the purpose of this study was to determine phylogenetic patterns in the evolution of phenotypic plasticity in response to vegetation shade (the ‘shade avoidance’ syndrome) in the annual plant Arabidopsis thaliana and its close relatives. Specifically, we asked the following questions: (i) Do A. thaliana and related species differ within or among clades in the magnitude and/or pattern of plasticity to shade? (ii) Are the phenotypic variance–covariance matrices (phenotypic integration) of these taxa plastic to the changes in light quality induced by the presence of a canopy? (iii) To what extent does the variation in uni- and multivariate plasticity match the phylogeny of Arabidopsis? In order to address these questions we grew individuals from six taxa of known phylogenetic relationship in a greenhouse under full sun and under a grass canopy. Taxa differed in the magnitude, but not in the pattern, of plasticities for all traits. At the univariate level, the late flowering species, A. pumila and A. griffithiana, as well as the late flowering Moscow ecotype of A. thaliana, showed greater plasticity for allocation to vegetative and reproductive meristems. At the multivariate level, several taxa displayed a very low stability of their variance–covariance structures to environmental change, with only one taxon sharing as many as three principal components across environments. We conclude that both univariate and multivariate plasticities to vegetation shade can evolve rapidly within a genus of flowering plants, with little evidence of historical constraints (phylogenetic inertia).

327 High Root-zone Temperature Effects on Floral Initiation in `Allstar' June-bearing Strawberry

Temperature, as a potential environmental stressor, interacts with photoperiod in floral initiation of June-bearing strawberries (Fragaria ×ananassa), such that high-temperature exposure can result in poor floral initiation. Our objectives were to examine the effects of various durations of high root-zone temperature on floral initiation and development and on vegetative growth and development. In a 1998 greenhouse experiment, hydroponically grown `Allstar' June-bearing strawberry plants were subjected day/night temperatures of 31/21 °C in the root zone for one, two, or three continuous periods (of ≈7 days), followed by exposure to 17 °C for the duration of the experiment. Control plants were raised at 17 °C in the root zone throughout the experiment. An additional temperature treatment was exposure to 31/21 °C in the root zone for two periods, each followed by a period at 17 °C. Plants were arranged in a randomized complete-block design with factorial treatments of duration of high root-zone temperature and harvest time. At the end of each period, plants were harvested and the apical meristems dissected for microscopic evaluation of vegetative and floral meristems and the stage of development of the primary flower. We observed floral initiation in all treatments after photoperiodic induction. However, exposure to 31/21 °C in the root zone during key periods of floral initiation in June-bearing strawberry may alter floral development.

Flower Bud Initiation in Primocane-fruiting Blackberry Germplasm

Scanning electron microscopy (SEM) and light microscopy (LM) were used to study the transition of meristems from vegetative to floral phase in erect primocane-fruiting (PF) blackberries [Rubus (Tourn.) L. subgenus Rubus] developed at the Univ. of Arkansas. Dormant root cuttings of A-1836 and APF-13 blackberries were dug from the field and planted on 28 Dec. 1996 and 1 Mar. 1997 to produce plants for use in a greenhouse study. In a field study, terminal buds of field-grown A-1836, APF-13, NC194, and summer-fruiting `Arapaho' were sampled on 21 Mar 1997 (before shoot emergence from soil), and then weekly from 14 to 28 May 1997. Flower bud primordia were first observed at five and six nodes of growth in greenhouse-grown A-1836 and APF-13 plants, respectively, 35 to 42 days after root cuttings were planted (DAP). Under field conditions, floral primordia were not observed until 21 May when A-1836 and APF-13 had at least 20 nodes of growth; NC194 did not differentiate floral structures until 10 July. The developmental patterns of the vegetative apical meristem in the PF selections, both field- and greenhouse-grown plants, were similar to those of `Arapaho'. Opening of the terminal flower of the inflorescence occurred 32 to 35 days after floral initiation in APF-13, and 8 to 10 days later on A-1836. Field-grown NC194 bloomed in late August. The first fruits of greenhouse-grown APF-13 were harvested 120 DAP. These findings demonstrate that PF blackberries form flower buds after a short period of vegetative growth.