Multiple Molecular Forms of Ficus glabrata Ficin. Their Separation and Relative Physical, Chemical, and Enzymatic Properties

Six of the proteolytic enzyme components of Ficus glabrata ficin have been isolated and shown to be chromatographically homogeneous. The molecular weights, the amino acid compositions, the electrophoretic and chromatographic behavior of the tryptic peptides, and the relative specificities of these 6 components have been determined. Within the experimental precision of the methods all 6 components are identical. They also have identical solubilities in sodium chloride and ammonium sulfate solutions. However, they are markedly different in their chromatographic properties. These multiple molecular forms of Ficus glabrata ficin may differ only in their conformational forms (conformers) or they may have minor differences in amino acid sequences which are sufficient to give different conformations and yet not be detected by the usual peptide mapping techniques. At the moment, we favor the latter possibility.     

Ultrastructure and Development of the Laticifers of Ficus carica L

Laticifers of Ficus caricaL. are of the branched, non-articulatedtype. They occur in the cortex and pith of the plant axis andpenetrate leaves and inflorescences. Observations were madeon laticifers located in shoot apices. Growing regions of laticifers undergo a sequence of ultrastructuralchanges. These are: (a) a pronounced increase in the vacuolarspace which divides the cytoplasm into separated masses; (b)a development of numerous vesicular structures in the cytoplasm.The vesicular structures are released into the vacuolar space.The whole process is accompanied by disintegration of cytoplasm.Apparently isolated masses of cytoplasm occur in the luminaof laticifer tips in sections taken from dormant apices. Itis assumed that these masses have a role in the initiation ofnew laticifer regions in the next growing season. Ficus caricaL., laticifers, ultrastructure, development differentiation     

Development of microsatellite markers in the common fig,Ficus carica L.

We present a new set of 15 polymorphic microsatellite primer sequences developed from Ficus carica L. The variability of specific microsatellite regions was assessed in wild population of figs from the northern Adriatic coast and all 15 primer pairs showed single‐locus amplification with a total of 65 alleles and an observed heterozygosity ranging from 0.285 to 0.863. The 15 new microsatellite loci represent a significant tool for population genetic structure studies and will be further used to investigate the origin and maintenance of genetic variation within and between populations of figs along the Adriatic coastal region.     

Growth and Respiratory Response of Fig (Ficus carica L. cv. Mission) Fruits to Ethylene

Growth in diameter of the fig (Ficus carica L. cv. Mission) fruit takes place in three distinct periods; two periods (I and III) of rapid growth are separated by a period (II) of slow growth. With respect to exposure to ethylene, the fruit exhibits a two phase response. Ethylene inhibits fruit growth in phase A (period I), the period of cell division, stimulates growth in early phase B (early period II), and stimulates both growth and ripening during the remainder of phase B (late period II and period III). The adverse effect of exogenous ethylene on fruits during phase A is thought to be due to inhibition of cell division. The gradual transition occurring in the response of fruits during phase B was interpreted in terms of carbohydrate level in the fruits.     

交联葡聚糖结合无花果蛋白酶的制备与性质

本文报道了将Sephadgx G-200与对β-硫酸酯乙砜基苯胺(SESA)首先醚化制备对氨基苯砜乙基交联葡聚糖(ABSE-Sephadex G-200),然后经重氮化固定无花果蛋白酶。固定化酶的活力回收达69％。BANA对该酶固定化过程中的活性变化有保护作用。天然酶与固定化酶都具有良好的耐热性,在69°～70℃,80min固定化酶较天然酶稳定。 用苯甲酰-DL-精氨酰-β-荼胺(BANA)为底物,在半胱氨酸存在下,测定了两种形式酶的动力学性质。在pH7.7的磷酸盐缓冲系统中,37℃,天然无花果蛋白酶的K_m=0.32mol/L;在间歇振摇下固定化酶的表观K′m=1.02mmol/L。最适pH无明显改变,均为pH7.7。     

无花果花芽分化与内源激素含量的关系

在‘布兰瑞克’无花果花芽分化形态学研究的基础上,对花芽分化期无花果新梢第7或第8节位花芽中的玉米素核苷(ZRs)、脱落酸(ABA)、赤霉素(GA1 3)、生长素(IAA)4种内源激素含量的变化进行了探讨。结果表明,在无花果花芽分化阶段,GA1 3和IAA初期含量较高,后快速下降,后期稳定在较低水平;ZRs和ABA在初期含量较低,后大幅提高,后期稳定在较高水平。可见,较高水平的内源ZRs、ABA和较低水平的内源GA1 3、IAA,以及较高的ABA/IAA、ABA/GA1 3、ZRs/GA1 3和ZRs/IAA比值有利于无花果花芽分化。     

Identification and characterization of microsatellite loci in the common fig (Ficus carica L.) and representative species of the genus Ficus

We developed microsatellites in fig (Ficus carica L.). A TC and TG‐enriched genomic library was screened, and after sequencing, primers were designed for 20 microsatellites. Eight primer pairs produced amplification products that were both interpretable and polymorphic in 14 fig cultivars and two French wild‐growing populations of F. carica (n₁ = 9 and n₂ = 10). Number of alleles per locus ranged from three to six. Except for one microsatellite locus, the observed heterozygosity was higher than the expected value. The F. carica microsatellites gave amplification products in 17 other Ficus species in 86% of the cases.     

Auxins and Gibberellin-like Substances in Parthenocarpic and Non-parthenocarpic Syconia of Ficus carica L., cv. King

In the King cultivar of fig, the first crop is parthenocarpic, develops on previous year's growth, and a series of supernumerary ovules develops within the original ovules. The second crop, formed on current-season's growth, requires fertilization. To determine whether the 2 crops differed in types, and in patterns of concentrations of total `free' auxins and acidic gibberellins, they were extracted from weekly fruit samples. Timing of the 3 peaks of total auxins and the single peak of gibberellins was identical in the 2 crops. The first auxin peak in both occurred at the end of fruit growth period I (first rapid growth period), the second shortly before the end of period II (period of slow growth), and the rise and fall in concentrations of the third peak accompanied the rise and fall of the fruit growth rate in period III. The end of period II was marked by the single gibberellin peak. Additional peaks before the first sampling dates, of auxins in the first crop, of gibberellins in the second, were indicated by high concentrations in the first samples and subsequent rapid decline. The same 4 individual auxins appeared in both crops. Auxins I and II were highest in concentration in the first total auxin peak of both crops. In the second peak of the first crop, auxins II and III were highest, whereas in that peak of the second crop auxins II and IV were highest. Qualitative differences in gibberellins occurred in the 2 crops. In general, auxin concentrations were higher in the first than the second crop, and gibberellin concentrations higher in the second. High concentrations of gibberellins coincided with low ones of auxins, and vice versa.     

Factors affecting in vitro regeneration of Ficus carica L. and genetic fidelity studies using molecular marker

Journal of Plant Biochemistry and Biotechnology - A facile method for regeneration of fig (Ficus carica L.) is in demand given the inability of the varieties having persistent type fruiting habit...     

Use of morphological traits and microsatellite markers to characterize the Tunisian cultivated and wild figs (Ficus carica L.)

We used 8 morphological traits and 17 simple sequence repeats loci to characterize 71 cultivated and wild Tunisian fig trees (Ficus carica L.). Significant morphological differences were inferred from leaf traits. The statistical analysis showed two major fig groups that indicated a common morphological basis. A total of 74 SSR alleles was revealed, defining 63 unique multilocus genotypes indicating a substantial genetic diversity. Based on multilocus SSR genotypes an identification key was established using MFC30, MFC3, MFC11 and MFC19 loci to identify figs. Analysis of variance components of linkage disequilibrium shown that among 136 pairs of loci, 32 present a significant gametic disequilibrium. The parameter D′²_IS (0.1284) was greater than D′²_ST (0.0079), a pointer of close to zero variance in total simple, and consequently the more pronounced independence of the 17 SSR loci. The majority of Ohta's variance components of linkage disequilibrium followed a pattern caused by genetic drift or a non-systematic disequilibrium profiles and natural selection occurs only for LMFC24-MFC8 pair loci in cultivated figs. Our results suggest that the morphological and SSR markers are suitable to characterize figs and should be recommended in conservation management strategy.     

Conversion of Tubercidin to Toyocamycin: Some Properties of Tubercidin Derivatives (Nucleosides and Nucleotides. 47)1

Abstract

A facile method of conversion of tubercidin to the 5-methyl derivative and toyocamycin is described. The NMR and CD spectra of 5- and 6-substituted tubercidins are presented. These data show that the 6-substituted tubercidins are in the syn-conformations in solution.     

Isolation and Some Properties of the Enzyme That Transforms Eremofortin C to PR Toxin

PR toxin and eremofortin C are secondary metabolites of Penicillium roqueforti. The chemical structures of these two compounds are closely related to each other and differ only by an aldehyde and an alcohol group at the C-12 position. In an effort to better understand the biosynthesis of PR toxin, we discovered the enzyme of P. roqueforti that is responsible for the transformation of eremofortin C to PR toxin. The maximum activity of the enzyme in the culture medium was found to occur on day 13, which corresponded to the maximal production of PR toxin in the medium. The enzyme was isolated and purified from the culture medium and the mycelium of the fungus, respectively, through a procedure involving ammonium sulfate fractionation and DEAE-cellulose chromatography. The specific activity increased 20- and 8-fold, respectively, and the yield was 33.3 and 21.6%, respectively, for the enzyme from the medium and mycelium. The optimal pH for the enzyme reaction was ca. pH 5.6. The enzyme reaction was temperature dependent. The rates followed a linear time course when it catalyzed the transformation at 30°C and decayed with time when reacted at higher temperatures. At 100°C, the enzyme activity was completely lost. The K_m and V_max of the enzyme as determined at 30°C were 0.02 mM and 4.0 μmol/min per mg, respectively. The molecular weight of the enzyme was estimated by gel filtration on a high-pressure liquid chromatography I-250 protein column to be ca. 40,000.     

盐胁迫下无花果细胞质膜和液泡膜H+-ATPase活性对脯氨酸积累的影响

盐胁迫降低无花果振荡培养细胞培养液pH ,添加质膜H ATPase活性抑制剂Na3VO4 则抑制盐诱导的培养液pH下降 ,表明盐诱导培养液pH下降主要是细胞质膜H ATPase活性增加的结果。NaCl处理提高活体细胞质膜H ATPase活性 ,而降低膜微囊H ATPase活性。培养液中添加Na3VO4 5 0 μmol/L完全抑制盐胁迫下无花果细胞游离脯氨酸积累 ,但添加更高浓度Na3VO4 ,则提高细胞液泡膜H ATPase活性 ,同时Na3VO4 抑制脯氨酸积累的效应下降 ,暗示盐胁迫下无花果细胞质膜和液泡膜H ATPase共同参与细胞质pH调节 ,影响游离脯氨酸积累。     

Some Physicochemical Properties and Substrate Specificity of Acid Protease of Rhodotorula glutinis K-24

Some physicochemical properties, amino acid composition, and substrate specificity of the acid protease of Rhodotorula glutinis K-24 were determined. The molecular weight was estimated to be about 30,000 by the sedimentation equilibrium method. This value also coincided with the data obtained from Andrews’s method.

The isoelectric point was pH 4.5, and the amino terminal amino acid was identified to be alanine. The enzyme contained 14.5% of nitrogen and was composed of 285 residues of amino acid. Substrate specificity toward synthetic peptides was similar to that of pepsin, but its activity was considerably weak.

The enzyme was inactivated by diazoacetyl glycine ethylester, p-bromophenacyl bromide, et al., which attacked the active center of pepsin.     

The Occurrence and Some Properties of a New Filamentous Component of Plant Extracts

The electron microscopic image and some properties of a filamentouscomponent of plant extracts are described. The filaments are3·5 nm in diameter, non-rigid and present a beaded appearancewhen positively stained with uranyl and lead salts. They arepresent in the microsomal supernatant fractions of a varietyof plants, and can be obtained in extracts of fresh tissuesor of acetone powder preparations. Specific tissues, successfullyextracted for filaments, include phloem, xylem and corticalparenchyma. They are not present in extracts of cell wall preparationsor of hair cells of cotton bolls. An outstanding characteristicof these filaments is their great stability to a wide varietyof treatments which includes variation in temperature, ionicenvironment, pH and the presence of urea, thiol reagents orthe detergent Nonidet. Lateral aggregation of the filamentsis evident below pH 3·0 and above pH 8·0. Althoughcellulase was the only enzyme of those tested which digestedthe filaments, it is unlikely, for several reasons, that thefilaments are a form of pure cellulose. Hydroxyproline was presentin all filament fractions after partial purification by variousmethods. The filaments are discussed in relation to known fibrillarcomponents of plant cells.     

Assessment of genetic stability on in vitro and ex vitro plants of Ficus carica var. black jack using ISSR and DAMD markers

Molecular Biology Reports - Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It...     

Indirect regeneration of Ficus carica by the TCL technique and genetic fidelity evaluation of the regenerated plants using flow cytometry and ISSR

Plant Cell, Tissue and Organ Culture (PCTOC) - Calcium-dependent protein kinases (CDPKs), as an important calcium sensor in plants, are widely involved in the signal transmission process of growth...