Effects of phytosulfokine-α on growth and tropane alkaloid production in transformed roots of Atropa belladonna

Phytosulfokine (PSK)- is a sulphated pentapeptide, isolated fromthe medium of cultured Asparagus officinalis mesophyllcells, that promotes cell proliferation. It is a putative key factor inconditioned medium required for the growth of low-density plant cell cultures.The present study investigates the effect of PSK- on growth and tropanealkaloid production in Atropa belladonna hairy rootstransformed with Agrobacterium rhizogenes (MAFF 03-01724). Although the growth rates ofhairy roots cultured in medium with orwithout PSK- for 4 weeks did not show any differences, the productivityof tropane alkaloids, especially of hyoscyamine, was enhanced by10^–7 or 10^–8 M PSK-. Inaddition, the content of tropane alkaloids in transformed roots treated withPSK- was 1.4 times higher than that of untreated roots after 4 weeks ofculture. The time course of growth and tropane alkaloid production inAtropa belladonna transformed roots suggested thatPSK- influenced the growth of transformed roots during the activegrowingphase, but not tropane alkaloid production.     

RFLP mapping of three new loci for resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) in barley

Three new major, race-specific, resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) were identified in three barley lines, RS42-6*O, RS137-28*E, and HSY-78*A, derived from crosses with wild barley (Hordeum vulgare ssp. spontaneum). The resistance gene origining from wild barley in line RS42-6*O, showed a recessive mode of inheritance, whereas the other wild barley genes were (semi)-dominant. RFLP mapping of these three genes was performed in segregating F₂ populations. The recessive gene in line RS42-6*O, was localized on barley chromosome 1S (7HS), while the (semi)-dominant genes in lines RS137-28*E, and HSY-78*A, were localized on chromosomes 1L (7HL) and 7L (5HL), respectively. Closely linked RFLP clones mapped at distances between 2.6cM and 5.3 cM. Hitherto, specific loci for powdery mildew resistance in barley had not been located on these chromosomes. Furthermore, tests for linkage to the unlocalized resistance gene Mlp revealed free segregation. Therefore, these genes represent new loci and new designations are suggested: mlt (RS42-6*O), Mlf (RS137-28*E), and Mlj (HSY-78*A). Comparisons with mapped QTLs for mildew resistance were made and are discussed in the context of homoeology among the genomes of barley (H-vulgare), wheat (Triticum aestivum), and rye (Secale cereale). Duplications of RFLP bands detected in the neighbourhood of Mlf and mlt might indicate an evolutionary interrelationship to the Mla locus for mildew resistance.     

Genetic analysis of brown planthopper resistance in twenty varieties of rice,Oryza saliva L.

Summary The inheritance of resistance to brown planthopper, Nilaparvata lugens (Stol.), of 20 rice cultivars was studied. Single dominant genes that are allelic to Bph 3 condition the resistance in cultivars Ptb 19, Gangala (Acc. 7733), Gangala (Acc. 15207), Horana Mawee, Kuruhondarwala, Mudu Kiriyal and Muthumanikam. Single recessive genes that are allelic to bph 4 govern the resistance in cultivars Gambada Samba, Heenhoranamawee, Hotel Samba, Kahata Samba, Kalukuruwee, Lekam Samba, Senawee, Sulai, Thirissa and Vellai Illankali. The resistance in Ptb 33, Sudu Hondarwala, and Sinna Sivappu is governed by one dominant and one recessive gene which segregate independently of each other. The dominant resistance genes in these cultivars appear allelic to either Bph 1 or Bph 3. Similarly, the recessive genes in these cultivars seem allelic to either bph 2 or bph 4. Further investigations are needed to conclusively determine the allelic relationships of resistance genes in Ptb 33, Sudu Hondarwala and Sinna Sivappu.     

Structures of asparagine linked oligosaccharides of immunoglobulins (IgY) isolated from egg-yolk of Japanese quail

Structures of the Asn linked oligosaccharides of quail egg-yolk immunoglobulin (IgY) were determined in this study. Asn linked oligosaccharides were cleaved from IgY by hydrazinolysis and labelled withp-aminobenzoic acid ethyl ester (ABEE) afterN-acetylation. The ABEE labelled oligosaccharides were then fractionated by a combination of Concanavalin A-agarose column chromatography and anion exchange, normal phase and reversed phase HPLC before their structures were determined by sequential exoglycosidase digestion, methylation analysis, HPLC, and 500 MHz¹H-NMR spectroscopy. Quail IgY contained only neutral oligosaccharides of the following categories: the glucosylated oligomannose type (0.6%, Glc1-3Glc1-3Man₉GlcNAc₂; 35.6%, Glc1-3Man_7–9GlcNAc₂). oligomannose type (15.0%, with the structure Man_5–9GlcNAc₂) and biantennary complex type with core structures of-Man1-3(-Man1-6)Man1-4GlcNAc1-4GlcNAc (9.9%),-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4GlcNAc (25.1%) and-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4(Fuc1-6)GlcNAc (11.4%). Although never found in mammalian proteins, glucosylated oligosaccharides (Glc₁Man_7–9GlcNAc₂) have been located previously in hen IgY.Abbreviations IgG, IgM, IgA, IgY immunoglobulin G, M, A and Y, respectively - ABEE p-aminobenzoic acid ethyl ester     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

RNA polymerase sigma-related proteins in Escherichia coli: detection by antibodies against a synthetic peptide

Summary Antibodies were raised against a synthetic tetradecameric peptide with an amino acid sequence, DLIQEGNIGLMKAV, which corresponds to the most highly conserved region of bacterial RNA polymerase factors. In a Western-blot analysis of total Escherichia coli proteins, the antiserum reacted specifically with at least three proteins with apparent molecular weights of 75 kDa, 27 kDa and 23 kDa, in addition to the known factors (⁷⁰ and ³²). The majorities of ⁷⁰ and ³² were recovered as associated forms with the RNA polymerase on glycerol gradient centrifugation, while the other cross-reacting proteins were not. Unambiguous evidence was obtained which indicated that the intracellular level of ³² increased rapidly upon heatshock, at least in the strain containing high copy numbers of the rpoH gene.     

Glycerol-, inositol-, and reducing end hexose-containing oligosaccharides in human urine

Ten previously unreported oligosaccharides have been purified from the urines of human subjects using a combination of gel filtration, ion exchange, and thin-layer chromatographies. Their structures were determined by direct probe mass spectrometry, methylation analysis, and proton NMR spectroscopy of the permethylated oligosaccharide alditols.On the basis of composition, the oligosaccharides could be divided into three groups. Five oligosaccharides containing glycerol were characterized as glucosyl1-1glycerol; glucosyl1-1glycerol; galactosyl1-1glycerol; glucosyl-1-1(fucosyl-1-2)glycerol and/or fucosyl-1-1(glucosyl-1-2)glycerol; and glucosyl-1-1(galactosyl-1-2)glycerol or galactosyl-1-1(glucosyl-1-2)glycerol. Four inositol-containing oligosaccharides were characterized as galactosyl1 (fucosyl1)inositol,N-acetylgalactosaminyl1 (fucosyl1)inositol, fucosyl1-2galactosyl1 (N-acetylgalactosaminyl1)inositol and fucosyl1-2galactosyl1-4-N-acetylglucosaminyl1(N-acetylgalactosaminyl1)inositol. Finally, galactosyl1-3(fucosyl1-2)galactosyl1-6galactosyl1-4(fucosyl1-3)glucose, an oligosaccharide with glucose at its reducing end, was tentatively identified. The significance and possible origins of the carbohydrate structures are discussed.     

Inheritance of callus formation ability in anther cultures of rice,Oryza sativa L.

Summary Inheritance of ability to form callus in rice anther culture was studied using the diallel technique. Anthers containing uninucleate microspores from two japonica cultivais (Minehikari and Taipei 309), two indica cultivars (Mingolo and Suweon 290), and 12 F₁'s of the diallel crosses involving these four parents were cultured on Chaleffs R₂ medium and evaluated for callus induction. The parents showed significant differences in anther callus formation, from 41.9% (Taipei 309) to 0% (Suweon 290). Callus induction ability was inherited as a recessive character conditioned by a single block of genes. Additive gene effects were predominant. The japonica types seemed to be good combiners for callus induction. The order of dominance among the four parents was Suweon 290, Mingolo, Minehikari and Taipei 309.     

Post-translational modifications of α-tubulin in<Emphasis Type="Italic"> Zea mays</Emphasis> L. are highly tissue specific

To further understand post-translational modifications (PTMs) of plant -tubulin, post-translationally modified -tubulin isoforms from selected tissues of Zea mays L. were examined using two-dimensional electrophoresis and immunoblotting. Except for polyglycylated tubulin, tyrosinated, detyrosinated, acetylated and polyglutamylated -tubulin isoforms were all present in maize tissues. Tyrosinated -tubulin was the predominant variant in all cases, with isoforms 1–4 (5) being the most common components. Leaves exhibited a striking difference in PTM patterns of -tubulin isoforms compared to other tissues examined. In leaves, several major specific isoforms were highly modified by detyrosination, acetylation and polyglutamylation. In pollen and anthers, only the most abundant isoform 3 was acetylated to an appreciable extent, and no acetylated isoform was found in roots. Similarly, in pollen, anthers and roots, only 3 was appreciably polyglutamylated. Additionally, a detyrosinated isoform 6 was present in anthers and in leaves, while the tyrosinated isoform 6 seemed to be pollen specific. These results indicate that certain types of PTM of plant -tubulin preferentially occur in a tissue-specific way.Abbreviations 1-, 2-D one-, two-dimensional - MT microtubule - PTM post-translational modification     

Alfalfa (Medicago sativa) Somatic Embryogenesis: Genetic Control and Introduction of Favourable Alleles into Elite Argentinean Germplasm

The objectives of this work were to isolate regenerative genotypes from the alfalfa Canadian cultivar Rambler to perform the genetic analysis of somatic embryogenesis in this species and to introgress regeneration favourable alleles into Argentinean alfalfa elite germplasm. For regeneration inheritance studies, greenhouse established plants from a cross between a Rambler derived regenerative clone and a non-regenerative Argentinean clone as well as self-pollinated progenies of these clones were analyzed. The results showed that somatic embryogenesis is under the control of two complementary genes. The introgression of somatic embryogenesis favourable alleles was performed using two populations derived from crosses between regenerative clones, obtained in the first step of this work, and non-regenerative clones of Argentinean alfalfa elite populations. Twenty-seven plants obtained through this process showed somatic embryogenesis ability. Two of them, designated as INTA R1 and INTA R2, exhibited a high regeneration capacity as well as high regeneration multiplicity. Our results demonstrate that Rambler derived somatic embryogenesis favourable alleles are successfully expressed in Argentinean elite alfalfa genetic background rendering it suitable for tissue culture-based breeding methodologies.     

Correlations between in vivo resistance to Fusarium and in vitro response to fungal elicitors and toxic substances in carnation

Summary With the aim of ascertaining the existance of a correlation between in vivo resistance to Fusarium oxysporum f. sp. dianthi and in vitro response to fungal elicitors and toxic substances, phenylalanine ammonialyase and phytoalexin accumulation, on one hand, and resistance to culture filtrate, on the other, were assayed in in vitro cultures of three susceptible and four resistant Dianthus caryophyllus cultivars. Cultivars showing varying degrees of resistance in vivo either tolerated higher culture filtrate concentrations (Niki) or showed high PAL activity and phytoalexin production when treated with Fusarium elicitor (Duca), or responded positively to both treatments (Mei-Ling, Pulcino). No such responses were shown in tissue cultures of susceptible cultivars. The differential response to the fungal elicitor seemed to be highly specific as genetic differences between cultivars were not observed in tissue cultures treated with other biotic (Phytophthora infestans) and abiotic (HgCl₂) elicitors.Abbreviations FuCWC cell wall components from Fusarium oxysporum f. sp. dianthi race 2 - PhCWC cell wall components from Phytophthora infestans - PAL phenylalanine ammonia-lyase (EC 4.3.1.5)     

Production of nematocidal compounds by hairy root cultures of Tagetes patula L.

Summary Marigold (Tagetes patula L.) hairy roots induced by infection with Agrobacterium rhizogenes produced -terthienyl when grown in darkness, and an n-hexane extract of the roots showed nematocidal activity. Depending on the hairy root line used, the level of -terthienyl varied from 15 to 1268 g per g dry weight, a level that corresponded to 0.15 to 12.7-fold that in intact roots. Analysis by HPLC indicated that the nematocidal activity was due predominantly to -terthienyl. However, it is suggested that nematocidal compounds other than -terthienyl are present in hairy roots cultured in the dark for long periods or in the light.     

Dependence of nitrogen- and phosphorus-regulation of β-lactam antibiotic production byStreptomyces clavuligerus on aeration level

Interference with -lactam production inStreptomyces clavuligerus by ammonium and phosphate ions, normally observed with optimum levels of aeration, was eliminated by restriction of the air supply. Under such a restriction, ammonium slightly stimulated and phosphate markedly stimulated -lactam antibiotic production. These are rare examples of regulation reversal by an environmental modification.     

Characterization of groups of the zygomycete genusRhizopus

Rhizopus is a zygomycetous genus. Several species of this taxon may infect humans and lower animals. Seventeen isolates ofRhizopus species in three distinct morphological groups were studied: the stolonifer group (sporangiophores greater than 1 mm in height, sporangial diameters of 100–275 µm, branched rhizoids); the arrhizus group (sporangiophores greater than 1 mm in height, branched rhizoids, sporangial diameters of 100–240 µm); and the microsporus group (sporangiophores less than 0.8 mm in height, sporangial diameters less than 100 µm, simple rhizoids). Maximal growth temperatures were characteristic: the stolonifer group grew at 30°C, the arrhizus group grew at 36°C, and the microsporus group grew at 45°C. The DNA mol% G + C base composition of all isolates ranged from 34.9 to 40.2% Species within the three groups were grouped by DNA differences. The arrhizus group was most distinctive with a value of 34.9–36.3%; the stolonifer and microsporus groups had G + C values of 37.0–39.3% and 37.8–40.2%, respectively. Our research clarifies and defines the G-C values of the three important groups ofRhizopus species.     

Inheritance of the dwarf plant type in blackgram (Vigna mungo (L.) Hepper)

Summary The inheritance of the dwarf plant type was studied in blackgram (V. mungo (L.) Hepper). Type 9 has erect plant type with normal internode length. The mutant line, EMSD has reduced internode length. The F₁, F₂ and F₃ generations of a cross between Type 9 and EMSD and its reciprocal were studied. The extreme dwarf plant type appeared to be governed by a single recessive gene, dw ₁ dw ₁ with no cytoplasmic effect.Part of Ph.D. Thesis submitted by the first author     

Effect of 2-deoxyglucose, α-methylglucoside,and glucosamine on aflatoxin production by Aspergillus parasiticus

The effects of 2-deoxyglucose (2-DOG), -methylglucoside (-MG), and glucosamine (GA) on aflatoxin production by Aspergillus parasiticus were studied using conidia-initiated and replacement cultures. In conidia-initiated, 2-DOG, -MG, and GA supported varying amounts of growth when employed as sole carbon sources. In both conidia-initiated and replacement cultures, 2-DOG, but not -MG nor GA, as sole carbon sources support toxin formation. None of the compounds inhibited aflatoxin production when used in combination with glucose. It appears that neither 2-DOG, -MG, nor GA can be considered nonmetabolizable analogs of glucose in A. parasiticus.Abbreviations YES yeast extract sucrose - PMS peptone-mineral salts - 2-DOG L-deoxyglucose - -MG -methylglucoside - GA glucosamine     

The endomembrane system and mechanism of membrane flow in the green alga,Gloeomonas kupfferi (Volvocales,Chlorophyta) II. A cytochemical analysis

Summary Cytochemical analysis of the endomembrane system of the chlamydomonad flagellate,Gloeomonas kupfferi (Chlorophyta), reveals distinct compartmentalization. Phosphatase localization shows that: IDPase is located throughout all cisternae of the dictyosome and vesicles associated with the contractile vacuole. Other alkaline phosphatases like TPPase, ATPase and ITPase were localized within the trans-face cisternae and vesicles of the contractile vacuole. IMPase was localized at the plasmamembrane and not within the endomembrane system. Acid phosphatases, incl. CMPase, NADPase and -glycerophosphatase, were localized in vesicles emerging from the central terminus of the trans-face of the dictyosome and in the peripheral vacuolar network. Silver proteinate labeling was noted in the dictyosome, contractile vacuole and on the anterior plasmamembrane. A summary of endomembrane compartmentalization and a putative interpretation of membrane flow and economy are presented.Abbreviations ER endoplasmic reticulum - IDPase inosine 5-diphosphatase - ITPase inosine 5-triphosphatase - ATPase adenosine 5-triphosphatase - TPPase thiamine pyrophosphatase - CMPase cytidine 5-monophosphatase - NADPase -nicotinamide adenine diphosphatase - AcPase acid phosphatase     

A simple method for maintaining high multiplication of Narcissus shoot cultures in vitro

A simple method for stimulating and maintaining high in vitro multiplication of Narcissus shoot clump cultures was developed. Shoot clumps were subjected either to normal cutting where leaves were trimmed to 20 mm in length at the beginning of each culture passage or to severe cutting where shoot clumps were cut down to the basal plate region removing all green tissue. Severe cutting at the beginning of each culture passage initially doubled the leaf multiplication, compared to normal cutting, but the difference between cutting treatments declined in successive passages. The improvement in leaf multiplication was maintained when shoot clumps were subjected to severe cutting only at every other culture passage, with no cutting in the alternate recovery passages. In vitro multiplication was increased by severe cutting in all seven Narcissus cultivars which were tested.Abbreviations NAA-1 naphthylacetic acid - BAP benzylaminopurine     

Effect of stalk damage on growth and yield of certain maize cultivars by the maize stalk borer Chilo partellus

The effects of different degrees of stem tunneling caused by different levels of artificial infestation by the larvae of Chilo partellus (Swinhoe) on growth and yield of 4 maize cultivars were studied. The susceptible Inbred A suffered much more tunneling than ICZ1-CM, ICZ2-CM and Katumani Composite B. This tunneling influenced significantly the plant growth of all the four cultivars. There was a significant negative correlation between tunneling and yield for the cultivars Inbred A and Katumani Composite B, but such a relationship was not significant for ICZ1-CM and ICZ2-CM. Such differences in the relationships between tunneling and yield among the 4 cultivars are due to different degrees of tunneling caused by C. partellus larvae to the plants of these cultivars.

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Résumé L'étude a porté sur la croissance et la production avec différents niveaux de minage des tiges de 4 cultivars de maïs, lors de divers degrés de contamination artificielle par des chenilles de C. partellus. Le cultivar sensible Inbred A souffre plus des mines de C. partellus que ICZ1-CM, ICZ2-CM et Katumani composite B. Les mines influent significativement sur la croissance des 4 cultivars. Il y a une corrélation négative significative entre le minage des tiges et la production pour les cultivars Inbred A et Katumani composite B, mais la relation n'est pas significative pour ICZ1-CM et ICZ2-CM. Les différences entre le minage des tiges et la récolte des 4 cultivars sont dues aux différents degrés de minage des tiges de ces cultivars par C. partellus.