Floral Organ Abscission Research Articles

Ethylene-dependent and -independent processes associated with floral organ abscission in Arabidopsis.

Abscission is an important developmental process in the life cycle of the plant, regulating the detachment of organs from the main body of the plant. This mechanism can be initiated in response to environmental cues such as disease or pathogen, or it can be a programmed shedding of organs that no longer provide essential functions to the plant. We have identified five novel dab (delayed floral organ abscission) mutants (dab1-1, dab2-1, dab3-1, dab3-2, and dab3-3) in Arabidopsis. These mutants each display unique anatomical and physiological characteristics and are governed by three independent loci. Scanning electron microscopy shows delayed development of the flattened fracture plane in some mutants and irregular elongation in the cells of the fracture plane in other mutants. The anatomical observations are also supported by breakstrength measurements that show high breakstrength associated with broken cells, moderate levels for the flattened fracture plane, and low levels associated with the initial rounding of cells. In addition, observations on the expression patterns in the abscission zone of cell wall hydrolytic enzymes, chitinase and cellulose, show altered patterns in the mutants. Last, we have compared these mutants with the ethylene-insensitive mutants etr1-1 and ein2-1 to determine if ethylene is an essential component of the abscission process and find that although ethylene can accelerate abscission under many conditions, the perception of ethylene is not essential. The role of the dab genes and the ethylene response genes during the abscission process is discussed.

Molecular characterization of two highly homologous receptor-like kinase genes, RLK902 and RKL1, in Arabidopsis thaliana.

Receptor-like kinases (RLKs) constitute a large family of signal perception molecules. We characterized two highly homologous RLK genes, RLK902 and RKL1, in Arabidopsis. RLK902 and RKL1 showed a 75% amino acid sequence identity over their entire regions. In the RLK902 pro::GUS transgenic lines, GUS activity was strong in the root tips, lateral root primordia, stipules, and floral organ abscission zones, while the RKL1 promoter activity was dominant in the stomata cells, hydathodes and trichomes of young rosette leaves, and floral organ abscission zones. Neither the rlk902 mutant line, rkl1 mutant line nor rlk902/rkl1 double-knockout mutant line showed any significant phenotypes under normal growth conditions. These results suggest that RLK902 and RKL1 might mediate the signal transduction pathway in which at least one other complementary signaling pathway to these two RLKs might exist.

Effect of regulated overexpression of the MADS domain factor AGL15 on flower senescence and fruit maturation.

We have examined the effect of regulated overexpression of AGL15, a member of the MADS domain family of regulatory factors, on reproductive tissues. Using molecular and physiological markers, we show that constitutive overexpression of AGL15 in Arabidopsis leads to delay and down-regulation of senescence programs in perianth organs and developing fruits and alters the process of seed desiccation. Through genetic crosses, we show that the rate of water loss in the maturing seeds is dictated by the genetic composition and physiological state of the maternal tissue, rather than the embryo. To define the developmental time and/or place when senescence programs are most affected by elevated AGL15 levels, we expressed AGL15 under the control of various promoters. Expression during senescence or in abscission zone cells did not produce delays in floral organ senescence or abscission. Using a glucocorticoid-inducible expression system, we show that an increase in AGL15 levels around the time of flower opening is necessary to delay senescence and increase floral organ longevity.

Temporal and spatial expression of a polygalacturonase during leaf and flower abscission in oilseed rape and Arabidopsis.

During leaf abscission in oilseed rape (Brassica napus), cell wall degradation is brought about by the action of several hydrolytic enzymes. One of these is thought to be polygalacturonase (PG). Degenerate primers were used to isolate a PG cDNA fragment by reverse transcriptase-polymerase chain reaction from RNA extracted from ethylene-promoted leaf abscission zones (AZs), and in turn a full-length clone (CAW471) from an oilseed rape AZ cDNA library. The highest homology of this cDNA (82%) was to an Arabidopsis sequence that was predicted to encode a PG protein. Analysis of expression revealed that CAW471 mRNA accumulated in the AZ of leaves and reached a peak 24 h after ethylene treatment. Ethylene-promoted leaf abscission in oilseed rape was not apparent until 42 h after exposure to the gas, reaching 50% at 48 h and 100% by 56 h. In floral organ abscission, expression of CAW471 correlated with cell separation. Genomic libraries from oilseed rape and Arabidopsis were screened with CAW471 and the respective genomic clones PGAZBRAN and PGAZAT isolated. Characterization of these PG genes revealed that they had substantial homology within both the coding regions and in the 5'-upstream sequences. Fusion of a 1,476-bp 5'-upstream sequence of PGAZAT to beta-glucuronidase or green fluorescent protein and transformation of Arabidopsis revealed that this fragment was sufficient to drive expression of these reporter genes in the AZs at the base of the anther filaments, petals, and sepals.

Temporal and Spatial Expression of a Polygalacturonase during Leaf and Flower Abscission in Oilseed Rape and Arabidopsis

During leaf abscission in oilseed rape (Brassica napus), cell wall degradation is brought about by the action of several hydrolytic enzymes. One of these is thought to be polygalacturonase (PG). Degenerate primers were used to isolate a PG cDNA fragment by reverse transcriptase-polymerase chain reaction from RNA extracted from ethylene-promoted leaf abscission zones (AZs), and in turn a full-length clone (CAW471) from an oilseed rape AZ cDNA library. The highest homology of this cDNA (82%) was to an Arabidopsis sequence that was predicted to encode a PG protein. Analysis of expression revealed that CAW471 mRNA accumulated in the AZ of leaves and reached a peak 24 h after ethylene treatment. Ethylene-promoted leaf abscission in oilseed rape was not apparent until 42 h after exposure to the gas, reaching 50% at 48 h and 100% by 56 h. In floral organ abscission, expression of CAW471 correlated with cell separation. Genomic libraries from oilseed rape and Arabidopsis were screened with CAW471 and the respective genomic clones PGAZBRAN and PGAZATisolated. Characterization of these PG genes revealed that they had substantial homology within both the coding regions and in the 5′-upstream sequences. Fusion of a 1,476-bp 5′-upstream sequence ofPGAZAT to β-glucuronidase or green fluorescent protein and transformation of Arabidopsis revealed that this fragment was sufficient to drive expression of these reporter genes in the AZs at the base of the anther filaments, petals, and sepals.

Involvement of polyamines in the control of fruitlet physiological abscission in grapevine (Vitis vinifera)

The potential contribution of polyamines (PAs) in the regulation of physiologically induced fruitlet abscission was investigated in cuttings from two cultivars of Vitis vinifera L., Pinot noir (PN) and Merlot (MRT). Abscission was higher in MRT than in PN and was preceded by a decrease in free PA levels. This decline was more pronounced in inflorescences than in leaves of the sensitive cultivar. Soluble conjugated PA showed an opposite trend in both cultivars. This suggests a cause‐effect relationship between free and/or conjugated PA levels in floral organs and susceptibility to abscission. Spermidine (Spd), but not putrescine (Put) or diaminopropane, supplied at 0.5–1 mM to the nutritive medium prior to the anthesis, increased free and conjugated PA levels in the inflorescences and markedly inhibited abscission. α‐Difluoromethylarginine, an inhibitor of arginine decarboxylase, but not α‐difluoromethylornithine, an inhibitor of ornithine decarboxylase, lowered PA levels and increased abscission. Treatment with cyclohexylamine or β‐hydroxyethylhydrazine as potent inhibitors of Spd synthase and PA oxidases, respectively, reduced the Spd and/or spermine levels and enhanced free Put in the inflorescences, inducing an increased abscission of floral organs shortly after anthesis. These data suggest that PAs, particularly Spd, could be involved in the regulation of grapevine fruitlet physiological abscission.

Fruit development is actively restricted in the absence of fertilization in Arabidopsis.

Flowering plants usually require fertilization to form fruit and seed and to initiate floral organ abscission in structures that do not contribute to the fruit. An Arabidopsis mutant that initiates seedless fruit without fertilization (fwf) or parthenocarpy was isolated and characterized to understand the factors regulating the transition between the mature flower and the initiation of seed and fruit development. The fwf mutant is fertile and has normal plant growth and stature. It sets fertile seed following self-pollination and fertilization needs to be prevented to observe parthenocarpy. The initiation of parthenocarpic siliques (fruit) was found to be dependent upon carpel valve identity conferred by FRUITFULL but was independent of the perception of gibberellic acid, shown to stimulate parthenocarpy in Arabidopsis following exogenous application. The recessive nature of fwf is consistent with the involvement of FWF in processes that inhibit fruit growth and differentiation in the absence of fertilization. The enhanced cell division and expansion in the silique mesocarp layer, and increased lateral vascular bundle development imply FWF has roles also in modulating silique growth post-fertilization. Parthenocarpy was inhibited by the presence of other floral organs suggesting that both functional FWF activity and inter-organ communication act in concert to prevent fruit initiation in the absence of fertilization.

HAESA, an Arabidopsis leucine-rich repeat receptor kinase, controls floral organ abscission

Abcission, the natural shedding of leaves, flowers and fruits, is a fundamental component of plant development. Abscission is a highly regulated process that occurs at distinct zones of cells that undergo enlargement and subsequent separation. Although some components of abscission, including accumulation of the hormone ethylene and cell wall-degrading enzymes, have been described, the regulatory pathways remain largely unknown. In this paper we describe a critical component required for floral organ abscission in Arabidopsis thaliana, the receptor-like protein kinase HAESA. Histochemical analysis of transgenic plants harboring a HAESA promoter:: beta-glucuronidase reporter gene and in situ RNA hybridization experiments show HAESA expression in the abscission zones where the sepals, petals, and stamens attach to the receptacle, at the base of pedicels, and at the base of petioles where leaves attach to the stem. Immunodetection, immunoprecipitation, and protein kinase activity assays reveal HAESA is a plasma membrane serine/threonine protein kinase. The reduction of function of HAESA in transgenic plants harboring an antisense construct results in delayed abscission of floral organs, and the severity of the phenotype is directly correlated with the level of HAESA protein. These results demonstrate that HAESA functions in developmentally regulated floral organ abscission.

Changes of Respiration Rate, Ethylene Evolution, and Abscisic Acid Content in Developing Inflorescence and Young Fruit of Olive (Olea europaea L. cv. Konservolia).

Simultaneous measurements of respiration, ethylene production, and abscisic acid (ABA) concentrations, as well as the growth parameters length, fresh weight (FW), and dry weight (DW) of olive (Olea europaea L. cv. Konservolia) inflorescence were carried out at short intervals (3-7 days) during the period from bud burst until the 3rd week after full bloom (AFB), when young fruit reached 8 mm in length. The axis of inflorescence elongated remarkably during the 3rd week after bud burst (ABB), massive bract shedding occurred during the 4th week ABB, full bloom (FB) was observed 7 weeks ABB, and massive floral organ abscission 1 week AFB. The results showed a continuous increase in inflorescence FW and DW from bud burst until 4 days before FB. Respiration rate, ethylene production, and levels of ABA were relatively high during the first 3 weeks ABB. After this period, respiration and ethylene followed a similar pattern of changes, inversely to that of ABA concentration. An accumulation of inflorescence ABA 6 and 4 days before FB was associated with the minimum values of respiration and ethylene production on the same dates. The sharp decrease in the ABA concentration during FB and 3 days later was followed by a high rise in ethylene and an increase in respiration rate, which both rose further 1 week AFB. The results suggest a possible correlation of ABA with the early stage of floral abscission, whereas ethylene production seems to be correlated with the terminal separatory activity in olive inflorescence abscission processes.Key Words. Abscisic acid-Abscission-Ethylene-Inflorescence-Olive-Respirationhttp://link.springer-ny.com/link/service/journals/00344/bibs/18n1p1.html

Structure and regulation of the Arabidopsis thaliana allene oxide synthase gene.

Allene oxide synthase (AOS) is encoded by a single intronless gene in Arabidopsis thaliana (L.) Heynh. The promoter region of the AOS gene exhibits, in addition to the clements of a minimal promoter and the presence of general enhancers, cis-elements that, in other promoters, are responsible for stress- and ethyleneresponsiveness. Arabidopsis thaliana and Nicotiana tabacum L. were transformed with a chimaeric gene consisting of a 1.9-kb 5'-upstream sequence and the first 95 nucleotides of the AOS coding sequence translationally fused to uidA encoding beta-glucuronidase (GUS). Using histochemistry, GUS activity was seen in older leaves, in the bases of petioles and in stipules, during the early stages of carpel development, in maturing pollen grains and at the base of elongated filaments, as well as in abscission-zone scars. A role for jasmonates in floral organ abscission is suggested by these findings. Furthermore, the AOS promoter was activated both locally as well as systemically upon wounding. Jasmonic acid, 12-oxophytodienoic acid and coronatine strongly induced GUS activity. This induction remained confined to the treated leaf when agonists were applied locally to a leaf, suggesting that neither jasmonic acid nor 12-oxophytodienoic acid are physiologically relevant components of the systemic wound signal complex. Rather, the data show that jasmonates behave as local response regulators produced at or around the sites of action in response to appropriate triggers of their synthesis.

POLLINATION REGULATION OF FLOWER DEVELOPMENT.

Pollination regulates a syndrome of developmental responses that contributes to successful sexual reproduction in higher plants. Pollination-regulated developmental events collectively prepare the flower for fertilization and embryogenesis while bringing about the loss of floral organs that have completed their function in pollen dispersal and reception. Components of this process include changes in flower pigmentation, senescence and abscission of floral organs, growth and development of the ovary, and, in certain cases, pollination also triggers ovule and female gametophyte development in anticipation of fertilization. Pollination-regulated development is initiated by the primary pollination event at the stigma surface, but because developmental processes occur in distal floral organs, the activity of interorgan signals that amplify and transmit the primary pollination signal to floral organs is implicated. Interorgan signaling and signal amplification involves the regulation of ethylene biosynthetic gene expression and interorgan transport of hormones and their precursors. The coordination of pollination- regulated flower development including gametophyte, embryo, and ovary development; pollination signaling; the molecular regulation of ethylene biosynthesis; and interorgan communication are presented.

Inflorescence Abscission in Olive: Anatomy and Histochemistry in Response to Ethylene and Ethephon

Naturally occurring postanthesis abscission in olive (Olea europaea L. Manzanillo) results in approximately 99% loss of flowers. Ethylene gas treatment induces sequential abscission of flowers, rachis branches and internodal sections, and peduncles of mature (expanded) inflorescences on shoot explants. Treatment with 2-chloroethyl phosphonic acid (ethephon) triggers sequential abscission of flowers and peduncles of mature inflorescences but only very infrequently rachises and never internodal sections. Immature (not fully expanded) inflorescences on shoot explants abscise intact in response to ethylene gas or ethephon. Sites of floral abscission on trees are limited to bracts, petals, individual flowers, and peduncles. With the exception of bracts, floral organs on trees do not abscise until after pollination. Naturally occurring abscission is preceded by plasmolysis of abscission zone cells, loss of cell wall materials as evidenced by changes in stain intensity, and lacunar cell separation. Abscission of floral organs of trees and of explants treated with ethephon or ethylene gas occurs at localized sites and is preceded by cell wall gelatinization and swelling (in ethephon-treated samples only), loss of cell wall and middle lamella pectins, lacunar cell separation, and senescence of cortical parenchyma cells. Phytotoxic symptoms, plasmolysis and cell senescence throughout all explant tissues occurred with ethephon treatments. All active abscission zone cells exhibit small size, isodiametric shape, dense cytoplasm, and starch accumulation. Cell division does not occur in any abscission zone with any treatment.

ウンシュウミカンの花器官の離脱時におけるデンプン粒の分布に関する解剖学的ならびに組織化学的研究

During the flowering stage of Wase satsuma mandarin, Citrus unshiu Marc. var. praecox Tanaka, the abscission layer formed at the junction of the base of the ovary and the filament or petal. The abscission layer was first evident one to three days after flowering. This layer was composed of 15 to 20 rows of cells in the constricted zone at the attachment points of filaments or petals and was first identified by its high affinity for iodine-potassium iodide. Cell separation occurred with rupturing of the cell walls.The abscission of floral organs was preceded by the deposition of starch grains in each of the cells which formed the abscission zone. After this starch accumulation was complete, no additional starch was deposited in the cells during abscission. Cellular separation occurred within the starch layer itself, and the starch was present in cells on both sides of the separation layer following completion of abscission. Subsequently, starch gradually disappeared from the parenchyma cells adjacent to the abscission layer.