RESUMEN
A novel genetic male sterile germplasm was developed by successively crossing of (C. annuum x C. chinense) x C. pubescens and by chemical mutagenesis in pepper. The sterile anthers showed morphological abnormalities, but pistils developed normally with fine pollination capability. We investigated fertility segregation through sib-crossing of the same strains and test crossing by male sterile plants with 6 advanced inbred lines. The results showed that male fertility in the pepper was dominant in the F1 generation and segregated at a rate of 3:1 in the F2 generation, suggesting that monogenic male sterility was recessive and conformed to Mendelian inheritance. Cyto-anatomy analysis revealed that microspore abortion of sterile anthers occurred during telophase in the microspore mother cell stage when tapetal cells showed excessive vacuolation, resulting in occupation of the loculi. The microspore mother cells self-destructed and autolyzed with the tapetum so that meiosis in pollen mother cells could not proceed past the tetrad stage.
Asunto(s)
Capsicum/genética , Infertilidad Vegetal/genética , Polen/citología , Capsicum/citología , Hibridación Genética , Mutagénesis , Polen/genética , TelofaseRESUMEN
Opening the porphyrin macrocycle of pheophorbide a and forming the primary fluorescent chlorophyll catabolites are key steps in the chlorophyll catabolism pathway. These steps are catalyzed by pheophorbide a oxygenase and red chlorophyll catabolite reductase (RCCR). In this study, a novel RCCR gene, CaRCCR, was isolated from the pepper (Capsicum annuum L.). The full-length CaRCCR complementary DNA is comprised of 1173 bp, contains an open reading frame of 945 bp, and encodes a 314-amino acid protein. This deduced protein belongs to the ferredoxin-dependent bilin reductase family. Amino acid sequence alignment showed that CaRCCR shared a high homology to other higher plant RCCR proteins. CaRCCR expression, as determined by quantitative real-time polymerase chain reaction, was higher in the leaves than the roots, stems, flowers, and immature fruits. CaRCCR expression was almost constant during all phases of leaf development. It was upregulated by abscisic acid, methyl jasmonate, and salicylic acid. Moreover, CaRCCR was induced by high salinity and drought stress treatments; it was also slightly regulated by Phytophthora capsici. Taken together, these results suggest that CaRCCR is involved in defense responses to various stresses.
Asunto(s)
Capsicum/enzimología , Capsicum/genética , Clorofila/metabolismo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Capsicum/efectos de los fármacos , Clonación Molecular , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Datos de Secuencia Molecular , Filogenia , Reguladores del Crecimiento de las Plantas/farmacología , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Proteínas de Plantas/química , Alineación de Secuencia , Análisis de Secuencia de ADN , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/genéticaRESUMEN
Twenty-two neutral O-linked oligosaccharides ranging from monosaccharides to octasaccharides were identified in bovine submaxillary-gland-mucin glycoprotein by a combination of liquid secondary-ion mass spectrometry, methylation analysis and 1H-NMR. Only five of these have been previously detected in bovine submaxillary-gland mucin although several have been described from other sources of mucin. The structures include short linear sequences 3-linked to N-acetylgalactosaminitol (GalNAcol) and branched structures based on either a GlcNAc(beta 1-6) [Gal(beta 1-3)]GalNAcol or GlcNAc(beta 1-6)[GlcNAc(beta 1-3)]GalNAcol core region. Oligosaccharides not previously characterised from any source were the disaccharide GalNAc alpha 1-6GalNAcol (GalNAc, N-acetylgalactosamine and the hexasaccharide GlcNAc(beta 1-6) [GalNAc(alpha 1-3)( Fuc (alpha 1-2)]Gal(beta 1-4)GlcNAc(beta 1-3)]GalNAcol (Fuc, L-fucose). Oligosaccharides of the blood-group-A type have not been detected previously in bovine submaxillary-gland mucin although their occurrence on bovine gastric-mucosal glycoproteins has been established by classical immunochemical studies.
Asunto(s)
Mucinas/metabolismo , Oligosacáridos/metabolismo , Glándula Submandibular/metabolismo , Sistema del Grupo Sanguíneo ABO/metabolismo , Animales , Secuencia de Carbohidratos , Bovinos , Cromatografía en Gel , Cromatografía de Gases y Espectrometría de Masas , Espectroscopía de Resonancia Magnética , Metilación , Datos de Secuencia MolecularRESUMEN
As part of the establishment of a data base for core and backbone sequences of O-linked oligosaccharides of human meconium glycoproteins, the minor tetra- to hepta-saccharides released from mild-acid treated blood group [corrected] H-active glycoproteins have been studied. These oligosaccharides are heterogeneous and difficult to isolate, and a t.l.c.-m.s. microsequencing procedure has been applied to the neoglycolipid derivatives, in conjunction with 1H-n.m.r. spectroscopy, methylation analysis, and mass spectrometry (m.s.) of native and methylated oligosaccharides. Among an array of oligosaccharides characterised are those having the branched beta-GlcNAc-(1----6)[beta-Gal- (1----3)]-GalNAcol core, and others with the following linear sequences not characterised previously from this source: beta-Gal-(1----3)-beta-GlcNAc-(1----3)-beta-Gal-(1----3)-GalNAcol, beta-Gal-(1----4)-beta-GlcNAc-(1----3)-beta-Gal-(1----3)-GalNAcol, alpha-GalNAc- (1----3)-beta-Gal-(1----3/4)-beta-GlcNAc-(1----3)-GalNAcol, beta-GlcNAc- (1----3)-beta-Gal-(1----3/4)-beta-GlcNAc-(1----3)- GalNAcol, beta-Gal-(1----3/4)-beta-GlcNAc-(1----3)-beta-Gal-(1----3/4)-beta- GlcNAc-(1----3)-beta-Gal-(1----3)-GalNAcol, and beta-GlcNAc-(1----3)-beta-Gal-(1----3/4)-beta-GlcNAc-(1----3)-beta-Gal- (1----3/4)-beta-GlcNAc-(1----3)-GalNAcol.
Asunto(s)
Glucolípidos/química , Glicoproteínas/química , Meconio/química , Oligosacáridos/química , Antocianinas , Secuencia de Carbohidratos , Cromatografía en Capa Delgada , Humanos , Recién Nacido , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Datos de Secuencia Molecular , Coloración y EtiquetadoRESUMEN
In the course of characterising neoglycolipid products derived from mucin oligosaccharide alditols after periodate oxidation and coupling by reductive amination to the aminolipid dipalmitoylglycerophosphoethanolamine, we have obtained evidence that oxidative cleavage occurs specifically at the C4-C5 bond of core N-acetylgalactosaminitol. Two lipid-linked fragments thus obtained from each oligosaccharide alditol are well resolved on thin-layer chromatography and can be sensitively analysed by liquid-secondary-ion mass spectrometry to assign the sequence and branching patterns of oligosaccharides linked at C6 and C3 to the N-acetylgalactosamitol. These conclusions have been reached from detailed studies of the neoglycolipid derivatives of several oligosaccharides (di- to hexasaccharides) which were isolated from human meconium and characterised previously by MS and NMR studies as the free alditols.
Asunto(s)
Glucolípidos/análisis , Oligosacáridos/análisis , Fosfolípidos/análisis , Alcoholes del Azúcar/análisis , Aminación , Conformación de Carbohidratos , Secuencia de Carbohidratos , Cromatografía en Capa Delgada , Humanos , Espectrometría de Masas , Meconio/análisis , Microquímica , Datos de Secuencia Molecular , Oxidación-Reducción , Ácido PeryódicoRESUMEN
The sensitivity of detection and extent of information on structure obtainable by liquid-secondary-ion mass spectrometry (l.s.i.-m.s.) of neoglycolipids on the conventional target probe and directly from the surface of silica plates following t.l.c. has been assessed. Neoglycolipids were derived from malto-oligosaccharides, chitin oligosaccharides, and a range of deoxyhexose-, hexose-, 2-acetamido-2-deoxyhexose-, and sialic acid-containing mammalian oligosaccharides by reductive amination using phosphatidylethanolamine dipalmitoate (PPEADP). Sub-pmol amounts of the maltopentaose-PPEADP derivative applied directly to the target probe provided information on molecular weight, whereas approximately 1 pmol was required when analysed on the silica gel t.l.c. plate. With a biantennary octasaccharide derivative, the sensitivity of detection was 20-50 times lower and the other oligosaccharides had intermediate sensitivities. Information on composition and sequence was obtained readily from fragment ions, using 5 pmol of the maltopentaose derivative and 50 pmol of the octasaccharide derivative on the target probe, and 50 and 200 pmol, respectively, on the silica gel chromatogram. The optimised conditions formed the basis for characterising the structures of the components of mixtures of oligosaccharides generated from glycoproteins.
Asunto(s)
Glucolípidos , Oligosacáridos , Conformación de Carbohidratos , Secuencia de Carbohidratos , Fenómenos Químicos , Química , Espectrometría de Masas , Datos de Secuencia Molecular , Fosfatidiletanolaminas , Sensibilidad y EspecificidadRESUMEN
Positive ion fast atom bombardment mass spectrometric sensitivities of seven amino acids and the corresponding di-isopropylphosphorylated derivatives were carefully compared. Results showed that an improvement in sensitivity by factors of 4-29, mostly above 10, were achieved after the derivatization. The chemical noise derived from glycerol matrix was also greatly reduced by this derivatization.