Reproductive Shoot Apex Research Articles

Auxin Response Factors promote organogenesis by chromatin-mediated repression of the pluripotency gene SHOOTMERISTEMLESS

Specification of new organs from transit amplifying cells is critical for higher eukaryote development. In plants, a central stem cell pool maintained by the pluripotency factor SHOOTMERISTEMLESS (STM), is surrounded by transit amplifying cells competent to respond to auxin hormone maxima by giving rise to new organs. Auxin triggers flower initiation through Auxin Response Factor (ARF) MONOPTEROS (MP) and recruitment of chromatin remodelers to activate genes promoting floral fate. The contribution of gene repression to reproductive primordium initiation is poorly understood. Here we show that downregulation of the STM pluripotency gene promotes initiation of flowers and uncover the mechanism for STM silencing. The ARFs ETTIN (ETT) and ARF4 promote organogenesis at the reproductive shoot apex in parallel with MP via histone-deacetylation mediated transcriptional silencing of STM. ETT and ARF4 directly repress STM, while MP acts indirectly, through its target FILAMENTOUS FLOWER (FIL). Our data suggest that – as in animals- downregulation of the pluripotency program is important for organogenesis in plants.

Sucrose synthesis in Unpollinated ovaries of pomegranate (Punica granatum L.), as well as, in reproductive and vegetative shoot apices

Sugar levels, the enzyme activity of UDP-Glucose Pyrophosphorylase (UGPase) and Sucrose Phosphate Synthase (SPS), key enzymes for sucrose synthesis, and the expression of their respective genes were examined in developing vegetative and reproductive shoot apices and in developing unpollinated ovaries of Pomegranate trees in order to detect differences possibly important for fruit production and plant vigor. The level of total sugars was higher in ovaries, followed by reproductive apices and vegetative apices, suggesting higher energy requirements in the reproductive organs. Moreover, the activities of SPS and UGPase were found to be elevated in the vegetative apices, indicating that sucrose synthesis is possibly differently regulated in comparison to the reproductive organs. Our results suggest that the sink power of each organ primarily regulates the levels of both transported (sucrose, mannitol) and non-transported (fructose, glucose) sugars. However, the endogenous production of sucrose may be a regulatory factor contributing to organ development.

The early inflorescence of Arabidopsis thaliana demonstrates positional effects in floral organ growth and meristem patterning

Key messageLinear modelling approaches detected significant gradients in organ growth and patterning across early flowers of the Arabidopsis inflorescence and uncovered evidence of new roles for gibberellin in floral development.Most flowering plants, including the genetic model Arabidopsis thaliana, produce multiple flowers in sequence from a reproductive shoot apex to form a flower spike (inflorescence). The development of individual flowers on an Arabidopsis inflorescence has typically been considered as highly stereotypical and uniform, but this assumption is contradicted by the existence of mutants with phenotypes visible in early flowers only. This phenomenon is demonstrated by mutants partially impaired in the biosynthesis of the phytohormone gibberellin (GA), in which floral organ growth is retarded in the first flowers to be produced but has recovered spontaneously by the 10th flower. We presently lack systematic data from multiple flowers across the Arabidopsis inflorescence to explain such changes. Using mutants of the GA 20-OXIDASE (GA20ox) GA biosynthesis gene family to manipulate endogenous GA levels, we investigated the dynamics of changing floral organ growth across the early Arabidopsis inflorescence (flowers 1–10). Modelling of floral organ lengths identified a significant, GA-independent gradient of increasing stamen length relative to the pistil in the wild-type inflorescence that was separable from other, GA-dependent effects. It was also found that the first flowers exhibited unstable organ patterning in contrast to later flowers and that this instability was prolonged by exogenous GA treatment. These findings indicate that the development of individual flowers is influenced by hitherto unknown factors acting across the inflorescence and also suggest novel functions for GA in floral patterning.

The RING Finger Protein NtRCP1 Is Involved in the Floral Transition in Tobacco (Nicotiana tabacum)

The transition from the vegetative phase to the reproductive phase is a major developmental process in flowering plants. The underlying mechanism controlling this cellular process remains a research focus in the field of plant molecular biology. In the present work, we identified a gene encoding the C3H2C3-type RING finger protein NtRCP1 from tobacco BY-2 cells. Enzymatic analysis demonstrated that NtRCP1 is a functional E3 ubiquitin ligase. In tobacco plants, expression level of NtRCP1 was higher in the reproductive shoot apices than in the vegetative ones. NtRCP1-overexpressing plants underwent a more rapid transition from the vegetative to the reproductive phase and flowered markedly earlier than the wild-type control. Histological analysis revealed that the shoot apical meristem of NtRCP1-overexpressing plants initiated inflorescence primordia precociously compared to the wild-type plant due to accelerated cell division. Overexpression of NtRCP1 in BY-2 suspension cells promoted cell division, which was a consequence of the shortened G2 phase in the cell cycle. Together, our data suggest that NtRCP1 may act as a regulator of the phase transition, possibly through its role in cell cycle regulation, during vegetative/reproductive development in tobacco plant.

A Molecular Framework for Auxin-Mediated Initiation of Flower Primordia

A classical role of the hormone auxin is in the formation of flowers at the periphery of the reproductive shoot apex. Mutants in regulators of polar auxin transport or in the auxin-responsive transcription factor MONOPTEROS (MP) form naked inflorescence "pins" lacking flowers. How auxin maxima and MP direct initiation of flower primordia is poorly understood. Here, we identify three genes whose expression is directly induced by auxin-activated MP thatfurthermore jointly regulate flower primordium initiation. These three genes encode known regulators offlower development: LEAFY (LFY), which specifiesfloral fate, and two AINTEGUMENTA-LIKE/PLETHORA transcription factors, key regulators of floral growth. Our study thus reveals a mechanistic link between flower primordium initiation and subsequent steps in flower morphogenesis. Finally, we uncover direct positive feedback from LFY to the auxin pathway. The auxin LFY module we describe may have been recruited during evolution to pattern other plant organ systems.

Anatomical characterisation of the foliar colleters in Myrtoideae (Myrtaceae)

Colleters are secretory structures that occur in vegetative or reproductive shoot apices of many botanical families. However, in the order Myrtales, reports of colleters have considered only external morphology. We therefore evaluated apical meristems of 52 species belonging to 17 genera from seven tribes of subfamily Myrtoideae (Myrtaceae), so as to analyse the incidence and morphological types of colleters. The samples were fixed for light and scanning electron microscopy. Histochemical tests were carried out on fresh and methacrylate-embedded material. Proteins of the colleter secretions were analysed by SDS-PAGE. We have classified and described the following three new colleter types: petaloid, conic and euryform. None of the species contained all three colleter types. The petaloid colleters were present in three tribes (Syzygieae, Melaleuceae and Lophostemoneae). The conic colleters were observed in three tribes (Leptospermeae, Myrteae and Melaleuceae) and the euryform type occurred in five tribes (Leptospermeae, Syncarpieae, Myrteae, Syzygieae and Melaleuceae). In the tribe Eucalypteae, we found no evidence of colleters. The presence of mucilaginous secretion that defines colleters was confirmed by histochemical tests, and no proteins were found in the secretion. The colleters in Myrtoideae may help clarify the phylogenetic relationships of the Myrtaceae family.

Structure, biochemistry and ecology of entomogenous galls in Selaginella Pal. Beauv. (Selaginellaceae) from India

We studied the structure of newly found entomogenous galls in three Indian species of Selaginella, and the biochemical changes during gall development as well as their seasonal abundances in 10 squares of 1.5 m2 under two differing habitat conditions, open and covered. Adult wasps (Cynipidae) initiate two types of gall formation by oviposition, spherical galls on vegetative shoots and elongated, club-shaped strobilar galls on reproductive shoots. Galls are anatomically and biochemically different from the unaffected shoots. Vegetative shoot apices bear more galls (84.6%) than reproductive shoot apices (15.4%). Gall frequency is significantly higher in covered (94.8%) than in open habitat (5.2%), and its seasonal peak occurs earlier in the former (43.1% during the rainy season) than in latter habitat (52.8% in autumn). We discuss the relationship between relative species density and gall seasonality as well as the possible role of certain environmental factors that make covered habitats more favorable to the gall inducer.

Distinct regulatory role for RFL , the rice LFY homolog, in determining flowering time and plant architecture

Activity of axillary meristems dictates the architecture of both vegetative and reproductive parts of a plant. In Arabidopsis thaliana, a model eudicot species, the transcription factor LFY confers a floral fate to new meristems arising from the periphery of the reproductive shoot apex. Diverse orthologous LFY genes regulate vegetative-to-reproductive phase transition when expressed in Arabidopsis, a property not shared by RFL, the homolog in the agronomically important grass, rice. We have characterized RFL by knockdown of its expression and by its ectopic overexpression in transgenic rice. We find that reduction in RFL expression causes a dramatic delay in transition to flowering, with the extreme phenotype being no flowering. Conversely, RFL overexpression triggers precocious flowering. In these transgenics, the expression levels of known flowering time genes reveal RFL as a regulator of OsSOC1 (OsMADS50), an activator of flowering. Aside from facilitating a transition of the main growth axis to an inflorescence meristem, RFL expression status affects vegetative axillary meristems and therefore regulates tillering. The unique spatially and temporally regulated RFL expression during the development of vegetative axillary bud (tiller) primordia and inflorescence branch primordia is therefore required to produce tillers and panicle branches, respectively. Our data provide mechanistic insights into a unique role for RFL in determining the typical rice plant architecture by regulating distinct downstream pathways. These results offer a means to alter rice flowering time and plant architecture by manipulating RFL-mediated pathways.

Flower primordium formation at the Arabidopsis shoot apex: quantitative analysis of surface geometry and growth

Geometry changes, especially surface expansion, accompanying flower primordium formation are investigated at the reproductive shoot apex of Arabidopsis with the aid of a non-invasive replica method and a 3-D reconstruction algorithm. The observed changes are characteristic enough to differentiate the early development of flower primordium in Arabidopsis into distinct stages. Primordium formation starts from the fast and anisotropic growth at the periphery of the shoot apical meristem, with the maximum extension in the meridional direction. Surprisingly, the primordium first becomes a shallow crease, and it is only later that this shape changes into a bulge. The bulge is formed from the shallow crease due to slower and less anisotropic growth than at the onset of primordium formation. It is proposed that the shallow crease is the first axil, i.e. the axil of a putative rudimentary bract subtending the flower primordium proper, while the flower primordium proper is the bulge formed at the bottom of this axil. At the adaxial side of the bulge, the second axil (a narrow and deep crease) is formed setting the boundary between the flower primordium proper and the shoot apical meristem. Surface growth, leading to the formation of the second axil, is slow and anisotropic. This is similar to the previously described growth pattern at the boundary of the leaf primordium in Anagallis.

Size and Activity of Shoot Apical Meristems as Determinants of Floret Number in Rice Panicles

The sink capacity (floret number per unit land area) is currently a serious constraint to grain yield production in japonica rice. The size and activity of the early reproductive shoot apex (incipient panicle) are potential determinants of the number of florets generated on the panicle. This hypothesis was tested using eight field-grown japonica rice cultivars (IR65598-112-2, IR65564-44-51, Nipponbare, Akenohoshi, Dobashi 1, Koshihikari, Kochihibiki and Nakateshinsenbon). Using morphometric microscopy, we found that the initial size of the reproductive shoot apex was highly correlated with the number of primary branches, but not with the number of florets per primary branch. The cell division activity of the early reproductive apex examined by in situ hybridization analyses using the histone H4 gene probe as a marker for DNA-replicating cells varied with the cultivar. Akenohoshi had twice as many DNA-replicating cells as Nipponbare and the cell division activity was highly correlated with the number of florets per primary branch, but not with the primary branch number. We concluded that the primary branch number was determined by the initial size of the reproductive apex, and that the floret number per primary branch was determined by the cell division activity in the following apex growth. This result provides the first evidence of a relationship between cell division activity and floret formation in the rice panicle.

Surface growth at the reproductive shoot apex of Arabidopsis thaliana pin-formed 1 and wild type.

With the aid of a non-destructive replica method and computational protocol, surface geometry and expansion at the reproductive shoot apex are analysed for pin-formed 1 (pin1) Arabidopsis thaliana and compared with the wild type. The observed complexity of geometry and expansion at the pin1 apex indicates that both components of shoot apex growth, i.e. the meristem self-perpetuation and initiation of lateral organs, are realized by the pin1 apex. The realization of the latter component, however, is only occasionally completed. The pin1 apex is generally dome-shaped, but its curvature is not uniform, especially later during apex ontogeny, when bulges and saddle-shaped regions appear on its periphery. The only saddle-shaped regions at the wild-type shoot apex are creases separating flower primordia from the meristem. Surface expansion at the pin1 apex is faster than at the wild type. In both pin1 and wild type the apex surface is differentiated into regions of various areal strain rates. In the pin1 apex, but not in the wild type, these regions correspond to the geometrically distinguished central and peripheral zones. Expansion of the central zone of the pin1 apex is nearly isotropic and slower than in the peripheral zone. The peripheral zone is differentiated into ring-shaped portions of different expansion anisotropy. The distal portion of this zone expands anisotropically, similar to regions of the wild-type apex periphery, which contact older flower primordia. The proximal portion expands nearly isotropically, like sites of flower initiation in the wild type. The peripheral zone in pin1 is surrounded by a 'basal zone', a sui generis zone, where areal strain rates are low and expansion is anisotropic. The possible relationships between the observed regions of different expansion and the various gene expression patterns in the pin1 apex known from the literature are discussed.

The liguleless2 gene of maize functions during the transition from the vegetative to the reproductive shoot apex.

During a maize plant's (Zea mays) development, the shoot apical meristem (SAM) generates an apex that proceeds through different phases: juvenile vegetative, adult vegetative and reproductive. During each phase the structures produced are distinguishable from structures produced during the other phases. In this paper, we demonstrate that the LIGULELESS2 (LG2) function is required for an accurate vegetative to reproductive phase transition. The maize gene liguleless2 (lg2) has been shown to encode a basic-leucine zipper (bZIP) protein and to function in narrowing the region from which the ligule and auricle develop in a typical maize leaf. Here we show that lg2 mutant plants can have reduced long tassel branches, extra vegetative leaves and extra husk leaves when compared to wild-type siblings. This indicates a role for the lg2 gene in the vegetative to reproductive phase transition of the shoot apex. We also discuss a potential role for the lg2 gene in general phase transition processes.

The Arabidopsis flowering-time gene LUMINIDEPENDENS is expressed primarily in regions of cell proliferation and encodes a nuclear protein that regulates LEAFY expression.

Mutations in the LUMINIDEPENDENS (LD) gene of Arabidopsis thaliana (L.) Heynh. (Arabidopsis) confer a late-flowering phenotype, indicating that LD normally functions to promote the floral transition. RNA and protein blot analyses, along with the analysis of transgenic plants containing a fusion between a genomic fragment of LD and the reporter gene uidA (GUS), indicate that LD is expressed primarily ipical proliferative regions of the shoot and root, including the shoot apical meristem and leaf primordia. Subcellular localization studies indicate that LD is a nuclear protein, consistent with its previously proposed transcriptional regulatory role. We have also found that in an apetala1 cauliflower (ap1 cal) background the ld mutation converts the reproductive shoot apex to a more vegetative state, a phenotype that is similar to that seen for the leafy (lfy) mutant. Furthermore, in situ hybridization analysis indicates that LFY levels are drastically reduced at the apex of ld ap1 cal plants after bolting. These data are consistent with the idea that at least one function of LD is to participate in the regulation of LFY.

Transgenic tobacco over-expressing a homeobox gene shows a developmental interaction between leaf morphogenesis and phyllotaxy.

The tobacco gene, NTH1, encodes a polypeptide of 326 amino acids and is a member of the class1 KN1-type family of homeobox genes. Expression of NTH1 has mainly been observed in vegetative and reproductive shoot apices, not observed in roots or expanded leaves. Over-expression of NTH1 in transgenic plants caused abnormal leaf morphology, consisting of wrinkling and curvature. Interestingly, the direction of leaf curvature tended to be conserved among almost all of the leaves in any given transformant. In transgenic plants exhibiting clockwise or anticlockwise phyllotaxy, leaves curved to the right or left, respectively, when looking from the shoot apex toward the base. Micro-surgical experiments demonstrated that the presence of the shoot apex is necessary for the development of leaf curvature, indicating that the order of formation of leaves on the stem (the generative spiral) affects leaf development. We found a correlation between the severity of leaf curvature and the value of the plastochron ratio, a parameter of phyllotaxy. Transformants with more severe phenotypes had larger plastochron ratios. From these findings, we discuss the possibility that an increase in the plastochron ratio, caused by over-expression of NTH1 in the shoot apex, may be involved in leaf curvature.

Expression of a rice homeobox gene causes altered morphology of transgenic plants.

We have isolated a cDNA clone encoding a homeobox sequence from rice. DNA sequence analysis of this clone, which was designated as Oryza sativa homeobox 1 (OSH1), and a genomic clone encoding the OSH1 sequence have shown that the OSH1 gene consists of five exons and encodes a polypeptide of 361 amino acid residues. Restriction fragment length polymorphism analysis has shown that OSH1 is a single-copy gene located near the phytochrome gene on chromosome 3. Introduction of the cloned OSH1 gene into rice resulted in altered leaf morphology, which was similar to that of the maize morphological mutant Knotted-1 (Kn1), indicating that OSH1 is a rice gene homologous to the maize Kn1 gene. RNA gel blot analysis has shown that the gene is primarily expressed in the shoot apices of young rice seedlings. This finding is supported by results of transformation experiments in which the 5' flanking region of the gene directed expression of a reporter gene in the shoot apex, particularly in stipules, of transgenic Arabidopsis. To elucidate the biological function of the OSH1 gene product, the coding region was introduced into Arabidopsis under the control of the cauliflower mosaic virus 35S promoter. Almost all transformants showed abnormal morphology. The typical phenotype was the formation of clumps of abundant vegetative and reproductive shoot apices containing meristems and leaf primordia, which did not form elongated shoots. Some transformants with a less severe phenotype formed elongated shoots but had abnormally shaped leaves and flowers with stunted sepals, petals, and stamens. The abnormal phenotypes were inherited, and the level of expression of the introduced OSH1 correlates with the severity of the phenotype. These findings indicate that the abnormal morphologies of the transgenic plants are caused by the expression of the OSH1 gene product and, therefore, that OSH1 is related to the plant development process.

Ontogeny of the reproductive shoot apex ofClethra barbinervis, especially on the superficial view

The structure and the ontogenetic process of the reproductive shoot apex forming a terminal inflorescence ofClethra barbinervis were examined, especially concerning the superficial view of the apex. The system of contact parastichies is 2+3 in phyllotaxis in the vegetative phase, changing to 5+8 for bract arrangement in the reproductive phase. At the same time the size of the apex is conspicuously enlarged. The size of the foliage leaf primordia in the vegetative phase is larger than that of the bract primordia in the reproductive phase. The radial cell files, which are clear in the vegetative shoot apex, are not recognizable at least in the early stage of the reproductive phase. The author proposes a close correlation between the appearance of the radial cell files, as well as the construction of the apical sectors, and the sizes of the shoot apex and leaf primordia. It may be proposed also that the construction of the apical sectors is closely correlated with the phyllotaxis.

Vegetative and reproductive shoot apex and root apex in someSaccharum species

Vegetative shoot apices of 7 varieties belonging to 4 species ofSaccharum exhibit a uniseriate tunica. Periclinal divisions are more the exception than the rule in them. The corpus is distinguishable into a small subapical initials zone and narrow flanking and pith initials zones. During the changeover of the shoot apex from the vegetative to the reproductive, it increases considerably in size and more tunica layers are laid down which persist even in the later stages of inflorescence development. The spikelet primordia originate from the second tunica layer and enlarge with cells contributed by the corpus. The root apex is of the common monocotyledonous type with three tiers of initials, one each for the stele and root cap, and a common one for the epidermis and cortex. It belongs to theZea type based on the pattern of root cap formation.

Evolutionary Trends in the Inflorescence of Angiosperms

The most primitive type of inflorescence in angiosperms is believed to be a leafy cyme. Examples from Magnoliaceae, Winteraceae, Annonaceae, Dilleniaceae, Ranunculaceae, Papaveraceae and Rosaceae indicate that even in these relatively primitive families the solitary flower is a derived condition. Determinate inflorescences are regarded as more primitive than indeterminate ones, a conclusion that is supported by examples from the Ranunculaceae, Papaverales, Rosaceae, Saxifragaceae, Apocynaceae, Campanulales and Violaceae. Trends of reduction and of amplification are both illustrated by examples. In the Compositae and Gramineae, two completely different patterns of development are superimposed upon each other, resulting in complex types of compound inflorescences. The ontogenetic development of inflorescences is related to the timing of the transformation of the shoot apex from the histological and physiological properties characteristic of its vegetative condition to the very different properties characteristic of the reproductive shoot apex. If this transformation coincides with or is immediately followed by the differentiation of the primordial floral appendages, a solitary flower or a simple leafy cyme is produced. In plants having very complex inflorescences, such as the capitulum of the Compositae and the panicle of the Gramineae, the transition occurs before the differentiation of separate floral primordia. In addition, reproductive apices that form complex inflorescences, containing bracts that are strongly differentiated from foliage leaves, are more strongly differentiated from vegetative apices than are reproductive apices that produce relatively simple, less well differentiated inflorescences. The distinctness of the inflorescence relative to the vegetative shoot depends to a large extent upon the abruptness of the transition of the shoot apex from the vegetative to the reproductive condition. The shape of the inflorescence is correlated with the orientation of mitoses in the primordial reproductive apex. The origin of indeterminate from determinate inflorescences has often been a two step process: reduction followed by amplification. These steps have been guided by alternating selective pressures: first for reduction of the length of the reproductive cycle in response to a shorter growing season; and second for increase in seed production in response to the selective pressures of a more favorable environment. Elongate racemes are most adaptive when the reproductive cycle is relatively long, but conditions of growth are less than optimum, because of low light, shortage of minerals, or other restrictions. Inflorescences that have repeated sympodial branching are highly flexible in their reaction to the environment, and so are particularly well adapted to regions where the favorable growing season fluctuates greatly from one year to another. The evolution of inflorescences illustrates very well two general properties of evolutionary trends: conservation of organization and adaptive modification along the lines of least resistance.