RESUMO
The purpose of the study was to assess the effects and the protective mechanism of konjac oligosaccharide (KOS) on the ulcerative colitis (UC) model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) in rats. KOS (1.0 and 4.0g/kg/day) was administered for 14days after the induction of colitis with TNBS. The status of the rats was assessed by morphological and biochemical methods. The effect of KOS on the colonic microflora was also assessed by studying the bacteria profile and short chain fatty acids (SCFAs) production in feces by standard culture techniques and gas chromatography, respectively. KOS administration improved rat weight, colonic length, damage score, structure of gut microbiota, production of SCFA, and reduced colon tissue levels of malondialdehyde (MDA), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß). Therefore, our results indicate that KOS is an anti-inflammatory and could be useful as a prebiotic to design functional foods for UC.
Assuntos
Amorphophallus/imunologia , Anti-Inflamatórios/uso terapêutico , Colite Ulcerativa/tratamento farmacológico , Colo/efeitos dos fármacos , Oligossacarídeos/uso terapêutico , Animais , Colite Ulcerativa/induzido quimicamente , Colo/microbiologia , Colo/patologia , Ciclo-Oxigenase 2/metabolismo , Modelos Animais de Doenças , Humanos , Interleucina-1beta/metabolismo , Masculino , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Sprague-Dawley , Ácido Trinitrobenzenossulfônico , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Amorphophallus konjac is an important economic crop widely used in health products and biomaterials. However, this monocotyledonous plant's production is seriously restricted by soft rot disease. Some Bacillus thuringiensis strains generate an endocellular acyl homoserine lactonase (AiiA), which has inhibitory effect on soft rot pathogen through disrupting the signal molecules (N-acylhomoserine lactones, AHL) of their Quorum Sensing system. The aim of our study is to obtain transgenic A. konjac expressing AiiA protein and exhibiting resistance to soft rot. But till now, there is not any report about exogenous gene transformation in A. konjac. In this research, an Agrobacterium-mediated genetic transformation system was constructed. An aiiA gene was synthesized according to the codon usage in A. konjac. Embryogenic callus was infected with the A. tumefaciens strain EHA105 harboring the plant transformation plasmid pU1301 plus synthesized aiiA gene. After antibiotics screening, 34 plants were obtained. PCR analysis showed that positive amplified fragments were present in 21 out of these 34 lines. Southern blot analysis indicated that aiiA gene had integrated into the genome of A. konjac. Western blotting demonstrated that the target protein of interest was reactive with the antibody against AiiA. Further disease resistance detection revealed that all of the tested transgenic A. konjac lines exhibited high resistance to soft rot bacteria Erwinia carotovora subsp. Carotovora (Ecc) SCG1. The protocol is useful for the quality improvement of A. konjac through genetic transformation.
Assuntos
Amorphophallus/genética , Amorphophallus/microbiologia , Bacillus thuringiensis/enzimologia , Hidrolases de Éster Carboxílico/genética , Imunidade Inata/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Amorphophallus/imunologia , Sequência de Bases , Southern Blotting , Western Blotting , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Genoma de Planta/genética , Dados de Sequência Molecular , Pectobacterium carotovorum/fisiologia , Doenças das Plantas/genética , Folhas de Planta/genética , Folhas de Planta/microbiologia , Plantas Geneticamente Modificadas , Reação em Cadeia da PolimeraseRESUMO
Reversed-phase high performance liquid chromatography was used to analyze water-soluble amino acids in the normal Amorphophallus Konjac, Amorphophallus albus, Amorphophallus bulbifer, and the soft rot Amorphophallus Konjac, to determine the relationship of the different soft-rot resistant Konjac varieties and the proportion and content of the multiple water-soluble amino acids. The results showed that there are remarkable differences in the content and proportion of water-soluble amino acids in different resistant varieties and the same variety of normal and diseased leaves of Amorphophallus. In this study, the bank of fingerprint 15 chromatogram was established and can be used to analyze the related characteristic peaks and the resistance of Amorphophallus.
