Cloning and expression analysis of CCoAOMT gene in fruit of ‘Dangshansuli'
Notice bibliographique
Résumé
[Objectives]Lignin is widely distributed in plant cell walls and confers strength,rigidity and impermeability,which are the principal components of stone cells in pear. Caffeoyl-CoA O-methyltransferase( CCoAOMT) plays an important role in the biosynthesis of lignin,and the 3-O-methylation of caffeoyl CoA leading to the formation of feruloyl Co A is catalyzed by the enzyme CCoAOMT.[Methods]In order to investigate the function of CCoAOMT in the synthesis of pear stone cells,15-year-old trees of Pyrus bretschneideri‘Dangshansuli'were used in this study,and we cloned Pb CCoAOMT gene and analyzed its biological information,and studied the gene expression at different stages during pear fruit development. These results contributed to providing some theoretical reference and practical basis to improve the quality of pear. Comparing CCoAOMT gene sequences associated with lignification obtained from from the NCBI database with pear genome database,the Pb CCoAOMT gene cDNA sequence involved in lignin biosynthesis was cloned by RT-PCR techniques. Gene structure analysis relied on GSDS. Sequence alignments and homology sequence searchers in databases were carried out using NCBI BLAST webpage. Deduced amino acid sequences were obtained by Primer Premier 5.0 software. Phylogenetic relationships among sequences were performed with MEGA 6.06. The molecular structure and physicochemical properties of proteins were analyzed by Prot Param. Hydrophobic analysis of deduced protein sequences was demonstrated by means of Prot Scale. Signal peptide of deduced protein sequences was predicted by Bio XM 2. 6. The transmembrane domain of deduced protein sequences was predicted by TMPRED. Secondary and tertiary structures of deduced protein sequences were aligned by SOPMA and SWISS-MODEL separately. For gene expression analysis,total genomic RNA was isolated from fruit samples collected at different stages during pearfruit development according to the CTAB method and c DNAs were generated following the manufacturers' recommendations to a final volume of 20 μL,gene quantifications were performed according to the manufacturers' instructions and Pyrus EFα1( EFα1,accession No.: AY338250) was used as internal gene to evaluate the RT-qPCR assays. [Results]The full-length cDNA of Pb CCoAOMT( Gen Bank accession No.: KJ577544) in pear was cloned. The sequence consists of 1 133 bp with an open reading frame of 744 bp,which encodes a polypeptide of 247 amino acids residues. Sequence alignment and phylogenetic analysis showed that CCoAOMT is a member of O-methyltransferases family and shared high similarity with CCoAOMT proteins from other species,especially with the highest similarity of 98.79% and phylogenetic relationship compared with the loquat. Real-time PCR results showed that the change of Pb CCoAOMT expression level was similar to that of stone cell content during fruit development,which showed a rise-down tendency.[Conclusions]Pb CCoAOMT gene was firstly isolated and characterized from pear,which may be involved in metabolism of lignin.
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,001 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».