Inhibition of Abscission of Bean Petiole Explants by Lepidimoide

The effect of lepidimoide on the process leading to abscission was studied in bean (Phaseolus vulgaris L. cv. Masterpiece) petiole explants. The assays, involving observations on the junction of the petiole of primary leaves and the pulvinus, were conducted in the light. Lepidimoide, at concentrations of 1 μm or higher, delayed the abscission process; however, the progression of abscission proceeded at normal rates, and complete abscission resulted. On the other hand indoleacetic acid inhibited the normal senescence resulting in greatly decreased abscission during the observation period. These observations show that lepidimoide only delays abscission, and the kinetics seem to indicate that lepidimoide and indoleacetic acid affect abscission through different mechanisms. Received March 1, 1996; accepted November 4, 1996     

Promotive Effect of C18-Unsaturated Fatty Acids on the Abscission of Bean Petiole Explants

The abscission-promoting effects of C₁₈-unsaturated fatty acidswere studied in bean (Phaseolus vulgaris L. cv. Masterpiece)petiole explants with the junction between the petiole and thepulvinus in the primary leaves in the light. Linolenic, linoleicand oleic acids promoted the abscission of the explants in thelight. Linolenic acid was the most effective among the compoundstested and its promotive effect was evident without any accompanyingincrease in the production of ethylene from the explants, ascompared with non-treated explants. Linolenic acid is easilyconverted to its hydroperoxide during the incubation with explants,as indicated by the formation of the conjugated diene and thegeneration of ethane. The production of ethylene from the explantstreated with linolenic acid was completely inhibited by theaddition of aminoethoxyvi-nylglycine (AVG), but large amountsof ethane were still generated. The promotive effect of linolenicacid was almost eliminated by the addition of scavengers offree radicals. Hydrogen peroxide and tert-butyl hydroperoxidepromoted abscission in the light. From these results, we concludedthat the abscission-promoting effect of linolenic acid are notmediated by the effect of ethylene but by the effect of itshydroperoxide, while the well-established pathway for the biosynthesisof ethylene from S-adenosylmethionine to ethylene, via 1-aminocyclopropane-l-carboxylicacid (ACC), was apparently operative. (Received May 1, 1991; Accepted July 10, 1991)     

Synthesis of Cellulase during Abscission of Phaseolus vulgaris Leaf Explants

When abscission in leaf explants from Phaseolus vulgaris, cultivar Red Kidney, was allowed to proceed while the explants were in ²H₂O, a 1.25% increase in the buoyant density of cellulase in a cesium chloride gradient was observed. These data indicate that the increase in cellulase activity during abscission is a result of the synthesis of new protein. Two differentially soluble forms of cellulase are present in the abscission zone. The form which is soluble only in a high salt buffer seems more closely related to the abscission process than the form which is soluble in dilute buffer. The correlation between changes in pull force and increase in cellulase activity and the effects of several hormones on cellulase activity are discussed.     

乙烯利对菜豆叶枕外植体脱落和离区纤维素酶合成的影响

以乙烯利处理菜豆叶枕外植体,能显著提高对抗的脱落及其离区纤维酶的活力。蛋白质和核酸合成的抑创剂环己酰亚胺和放线菌素Ｄ,对乙烯利促进的脱落与离区纤维素酶活力不仅有明显的抑制作用,而且具有严格的时间顺序。提示乙烯利促进脱落的生理效应与其诱导高区纤维素酶合成时基因表达的转录和翻译过程有密切关系。此外,外植体经乙烯利处理后再分别不同时间加ＩＡＡ,并根据测定其抑制乙烯利诱导的纤维素酶活力变化,与不同时间加放线菌素Ｄ的实验结果相同,推断ＩＡＡ对乙烯利促进脱落的拮抗作用可能是在乙烯利诱导纤维素酶合成的转录过程。     

Fine Structure and Cytochemistry of the Abscission Zone Cells of Phaseolus Leaves: I. Ultrastructural Changes occurring during Abscission

The fine structure of the separation zone cells from the basalabscission zone of bean leaves has been described. Examinationof these cells revealed that fracture occurred primarily asa result of the dissolution of the middle lamellar region ofthe walls, leaving intact cells on the two newly exposed fracturesurfaces. The cytoplasm and organelles within these cells appearedquite normal and showed no signs of autolysis or senescence.A comparison of the organelle numbers in these cells with cellsfrom a similar region of a control plant revealed large increasesin the number of Golgi bodies and the amount of rough endoplasmicreticulum. The significance of this finding is discussed inrelation to the secretion of cell wall hydrolysing enzymes whichare known to be produced in these cells.     

葡萄贮藏过程中落粒与离区酶活性变化及植物生长调节物质的关系

研究了无核白葡萄(Vitis vinifera L.)采后贮藏过程中离区纤维素酶、果胶酯酶(Pectinesterase,PE)、多聚半乳糖醛酸酶(polygalacturonase,PG)、脂氧合酶(lipoxygenase,LOX)和过氧化物酶(peroxidase,POD)活性的变化与落粒的关系及植物生长调节物质对其的影响。结果表明,葡萄在贮藏过程中,伴随浆果落粒的增加,离区纤维素酶、PG、LOX、POD活性升高,PE活性下降。离区纤维素酶、PG、LOX等酶的活性与葡萄落粒程度之间呈显著正相关。外源ABA和CEPA处理能增强离区纤维素酶、PG、LOX活性,促进落粒;GA3,IAA处理则能抑制离区纤维素酶、PG、LOX活性,减轻落粒。ABA对落粒的促进效应及GA3对纤维素酶活性和落粒的抑制效应尤为明显,表明GA3与ABA比值在葡萄采后落粒过程中起重要的作用。     

萌发绿豆子叶衰老期间蛋白质代谢的变化

萌发绿豆子叶自然衰老过程中可溶性蛋白质含量一直下降;从衰老开始到衰老前期,总游离氨基酸含量明显上升;但游离氨基酸各组分在子叶衰老期间的变化趋势并不相同。~3H-亮氨酸掺入蛋白质试验和多聚核糖体的相对量及其与总核糖体的比值(P/T)测定都证明在子叶衰老前期有蛋白质的新合成。子叶衰老期间。氨肽酶活性明显降低;而以酪蛋白为底物的蛋白水解酶活性却急剧上升,承担着催化蛋白质降解的主要功能。     

Protein Synthesis and Accumulation in Bean Cotyledons during Growth

Analysis of total protein, of specific proteins by gel electrophoresis and immunoelectrophoresis, and of protein synthetic activity in vitro confirmed that intense protein synthesis and accumulation occurred as the French bean (Phaseolus vulgaris L). seed grew from 12 to 20 millimeters. These techniques showed that there was no globulin-1 (G1) fraction (requiring high salt for solubility) present in 6-millimeter seeds, and only very small amounts were synthesized in seeds less than 9 millimeters long. The 7- to 9-millimeter stages represent a 2-day transition period over which genetic information for the G1 protein becomes actively expressed, accounting for at least 50% of all protein synthesized in this tissue during the following 14 days. At maturity, the electrophoretic analysis confirmed that G1 globulin was the major storage protein, representing some 50% of the dry seed protein. Cell-free protein synthesis assays, including immunoprecipitation of the in vitro products, clearly showed G1 polypeptides to be among the polysome-directed products.     

Seasonal Changes in the Levels of Some Cellular Components in the Abscission Zone of Coleus Leaves of Different Ages

The present study with intact petioles of Coleus of varyingages suggests an involvement of the phenomenon of mobilizationwith the abscission process. There was a gradual increase inthe levels of chlorophyll and nucleic acids and also solublenitrogen compounds in the proximal tissues with the approachof abscission with a concomitant decrease of these substancesin the distal tissues. This increase of metabolites in the proximaltissues was clearer in petioles at the first node than at thefourth node suggesting a lesser degree of metabolic activityof the proximal tissues in older petioles. The change of levelsof chlorophyll and nucleic acids and also of soluble nitrogenwas more marked in winter than in summer months. The application of NAA to petioles of the third node; just afterdeblading (i.e. in the auxin-inhibited stage 1), not only stoppedthe decline of nucleic acid levels (particularly RNA) at thedistal tissues but also caused an increase of nucleic acidsover the initial levels. This effect was more pronounced insummer than in winter months. If the initially inhibitory concentration of NAA was appliedin the auxin-promoted stage 2 of abscission, instead of an increase,there followed a quicker rate of decrease in nucleic acids (particularlyRNA). This effect was also more prominent in summer.     

Effect of Protein Synthesis Inhibitors, Auxin, and Gibberellic Acid on Abscission

Chloramphenicol, actinomycin D, and other inhibitors of protein synthesis promote abscission in several plant genera. Abscission is accelerated in species where an abscission layer is present, as well as in tissue where no abscission layer develops prior to abscission. The inhibitors promote abscission in species where cell division is reported to precede the separation processes as well as in tissues where no cell division is associated with the initiation of abscission. Indoleacetic acid (IAA) or auxin precursors, when applied with chloramphenicol and aclinomycin D, overcome the promotive effects of the inhibitors on abscission. These inhibitors apparently do not promote abscission through their effects on auxin precursor conversion, IAA transport, and IAA destruction in the petiole. IAA increases the incorporation of leucine-1-¹⁴C into a trichloroacetic acid precipitable fraction of the abscission zone under conditions where abscission is retarded. A low concentration of IAA which accelerates abscission, decreases incorporation of leucine into protein. Other promoters of abscission — chloramphenicol, d-aspartic acid, and gibberellic acid —also decrease the incorporation of leucine into the protein of the abscission zone. The data indicate that enzymes required for the degradative processes associated with abscission are already present in the abscission zone whereas a continuous synthesis of protein is required for the retention of the leaf.     

Abscission of Phaseolus and Impatiens Explants: Effects of Ionizing Radiation upon Endogenous Growth Regulators and de Novo Enzyme Synthesis

Stem-petiole explants from the lower pulvinus of the primary leaves of Phaseolus vulgaris L. cv. Red Kidney and from Impatiens sultani Hook cv. Scarlet Baby were exposed to varying dosages of γ-radiation. With bean, irradiation of 175 to 525 kiloroentgens (kR) significantly accelerated the onset of abscission with a maximum response at 175 to 280 kR. Higher dosages (beginning at 600-700 kR) usually prevented abscission. With Impatiens, 18 to 35 kR significantly accelerated both the onset of abscission and possibly the initial abscission rate; 350 kR cut the time to 100% abscission in half and substantially accelerated the initial abscission rate. Inhibition of abscission in Impatiens was not possible with the available dose rate (35 kR/hour).     

Epidermal Cells Do Not Contribute to Auxin-Induced Ethylene Production in Mung Bean Stem Sections

Auxin-induced and 1-aminocyclopropane-1-carboxylic acid (ACC)-dependentethylene production in mung bean (Vigna radiata [L] Wilczek)hypocotyl sections, from which epidermis had been removed, wasinvestigated. Ethylene production in hypocotyl sections withoutepidermis was induced by treatment with IAA, and also occurredfrom exogenously supplied ACC in the presence of 0.2 M mannitol.Isolated epidermal strips alone failed to produce substantialamounts of ethylene in response to IAA or from exogenous ACC.3,4-[¹⁴C]-Methionone was incorporated into both ACC and ethylenein peeled sections treated with IAA, but not in the isolatedepidermal strips. Radioactive ACC, however, was detected inthe epidermal strips separated from the unpeeled sections previouslyfed with 3,4-[¹⁴C]-methionine in the presence of IAA. We concludethat the Site of auxin-induced ethylene production is not inthe epidermis, but in other hypocotyl cells, and that epidermalcells lack the activity which converts ACC to ethylene. (Received January 28, 1985; Accepted May 4, 1985)     

Collagen-Induced Changes in the Pattern of Protein Synthesis of Fibroblasts

Cells were cultured on plastic, collagen fibrils or gelatin. General protein synthetic activity of cells did not show any significant, difference among the three substrates, whereas the pattern of protein synthesis was substrate-dependent. Profiles of protein synthesis (polypeptide maps) were obtained by subjecting two-dimensional autoradiograms of poly-acrylamide gel electrophoresis to a computer-assisted image analyzer. Major polypeptide spots expressed on gelatin were rather like those on plastic. Collagen fibrils caused significant changes in the polypeptides map. Fibroblasts on collagen fibrils produced 364 spots of polypeptides, 26% of which were synthesized specifically on collagen fibrils. The remaining was shared by cells on plastic and was categorized into three groups: (1) polypeptides whose synthesis was up-regulated by collagen fibrils (26% of the total); (2) polypeptides that were expressed equally on both plastic and collagen fibrils (51%); and (3) polypeptides down-regulated by'collagen fibrils (23%). A protein with molecular weight of 150 K and an isoelectric point (pI) of 7.3 was one of the collagen-induced and worthy of further analysis. This protein was found to change its pi depending upon the amounts of collagen fibrils and was shown to be located in the mitochondrial fraction.     

Transdifferentiation of Mature Cortical Cells to Functional Abscission Cells in Bean

Abscission explants of bean (Phaseolus vulgaris L.) were treated with ethylene to induce cell separation at the primary abscission zone. After several days of further incubation of the remaining petiole in endogenously produced ethylene, the distal two-thirds of the petiole became senescent, and the remaining (proximal) portion stayed green. Cell-to-cell separation (secondary abscission) takes place precisely at the interface between the senescing yellow and the enlarging green cells. The expression of the abscission-associated isoform of β-1,4-glucanhydrolase, the activation of the Golgi apparatus, and enhanced vesicle formation occurred only in the enlarging cortical cells on the green side. These changes were indistinguishable from those that occur in normal abscission cells and confirm the conversion of the cortical cells to abscission-type cells. Secondary abscission cells were also induced by applying auxin to the exposed primary abscission surface after the pulvinus was shed, provided ethylene was added. Then, the orientation of development of green and yellow tissue was reversed; the distal tissue remained green and the proximal tissue yellowed. Nevertheless, separation still occurred at the junction between green and yellow cells and, again, it was one to two cell layers of the green side that enlarged and separated from their senescing neighbors. Evaluation of Feulgen-stained tissue establishes that, although nuclear changes occur, the conversion of the cortical cell to an abscission zone cell is a true transdifferentiation event, occurring in the absence of cell division.