RESUMEN
Plant defensins are divided into 18 groups and are multifunctional proteins. The Zea mays defensin 1 (ZmDEF1) gene encodes the defensin 1 protein, which can inhibit alpha-amylase in the insect gut. In this study, the ZmDEF1 gene was transferred into two maize cultivars, LC1 and LVN99, to improve weevil resistance in maize. The recombinant ZmDEF1 protein was assessed for its ability to inhibit alpha-amylase in the gut of the larvae of the maize weevil (Sitophilus zeamais Motsch.). ZmDEF1 was cloned into a pBetaPhaso-dest vector, which harbours phaseolin, a seed-specific promoter, and the Agrobacterium tumefaciens strain C58 harbouring the pBetaPhaso-ZmDEF1 vector was used to transfer the ZmDEF1 gene into two maize cultivars using immature embryos. Transformed calluses were selected on selection media containing kanamycin. The stable integration of the ZmDEF1 transgene into the transgenic maize plant genome was confirmed using Southern blotting. The recombinant ZmDEF1 protein of approximately 10 kDa was expressed in three transgenic maize lines from the LC1 cultivar (C1, C3, and C5) and two transgenic maize lines from the LVN99 cultivar (L1 and L3). The ZmDEF1 transgenic efficiency based on the results of PCR, as well as Southern and Western blotting, was 1.32% and 0.82%, respectively, which depends on the genotypes of LC1 and LVN99. The recombinant ZmDEF1 protein inhibited the alpha-amylase activity of the maize weevil larvae, and its ability to inhibit alpha-amylase is 54.52-63.09% greater than the ZmDEF1 protein extracted from non-transgenic plants.
Asunto(s)
Defensinas/genética , Control Biológico de Vectores/métodos , Zea mays/genética , Agrobacterium tumefaciens/genética , Animales , Protección de Cultivos/métodos , Larva/metabolismo , Plantas Modificadas Genéticamente/genética , Semillas/genética , Transgenes/genética , Gorgojos/patogenicidad , Zea mays/metabolismo , alfa-Amilasas/antagonistas & inhibidoresRESUMEN
Chitinases play the key role in hydrolysis of chitin, a huge organic carbon reservoir on earth, into monomeric sugars and their eventual conversion into valuable chemicals and energy sources. The Lecanicillium lecanii strain 43H was used as the source for the Endochitinase gene without signal peptide (mchit1). This mchit1 gene was cloned and sequenced. The recombinant Endochitinase non signal peptide was overexpressed in Pichia pastoris X33 with a level of 2.048 U mL-1 culture supernatant. The molecular mass of the purified recombinant Endochitinase (rmchit1) without signal peptide was 43 kDa. Metal ions, detergents, and organic solvents tested indicated a significantly influence on rmchit1 activity. The obtained results demonstrated that signal peptides affect the yield expression, purification methods, recovery as well as the physicochemical properties of the enzyme.
Asunto(s)
Quitinasas/genética , Quitinasas/metabolismo , Hypocreales/enzimología , Pichia/crecimiento & desarrollo , Quitinasas/química , Clonación Molecular , Estabilidad de Enzimas , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Concentración de Iones de Hidrógeno , Peso Molecular , Pichia/genética , Pichia/metabolismo , Ingeniería de Proteínas , Señales de Clasificación de Proteína , Análisis de Secuencia de ADN , TemperaturaRESUMEN
Two new steroidal saponins, 26-O-ß-D-glucopyranosyl-22α-methoxy-5α-furost-25(27)-ene-1ß,3ß,26-triol 1-O-ß-D-glucopyranoside (1), and 26-O-ß-D-glucopyranosyl-22α-methoxy-furosta-5,25(27)-diene-1ß,3ß,26-triol 1-O-α-L-rhamnopyranosyl-(1â2)-ß-D-fucopyranoside (2) were isolated and elucidated from the roots of Cordyline fruticosa (L.) A. Chev. Their structures were established by interpretation of spectroscopic data (1 D and 2 D NMR) and mass spectrometry (HR-ESI-MS).
RESUMEN
Adrinandra megaphylla Hu is a medicinal plant belonging to the Adrinandra genus, which is well-known for its potential health benefits due to its bioactive compounds. This study aimed to assemble and annotate the chloroplast genome of A. megaphylla as well as compare it with previously published cp genomes within the Adrinandra genus. The chloroplast genome was reconstructed using de novo and reference-based assembly of paired-end reads generated by long-read sequencing of total genomic DNA. The size of the chloroplast genome was 156,298 bp, comprised a large single-copy (LSC) region of 85,688 bp, a small single-copy (SSC) region of 18,424 bp, and a pair of inverted repeats (IRa and IRb) of 26,093 bp each; and a total of 51 SSRs and 48 repeat structures were detected. The chloroplast genome includes a total of 131 functional genes, containing 86 protein-coding genes, 37 transfer RNA genes, and 8 ribosomal RNA genes. The A. megaphylla chloroplast genome indicated that gene content and structure are highly conserved. The phylogenetic reconstruction using complete cp sequences, matK and trnL genes from Pentaphylacaceae species exhibited a genetic relationship. Among them, matK sequence is a better candidate for phylogenetic resolution. This study is the first report for the chloroplast genome of the A. megaphylla.
Asunto(s)
Ericales/clasificación , Ericales/genética , Genoma del Cloroplasto , Genómica , Plantas Medicinales/clasificación , Plantas Medicinales/genética , Codón , Biología Computacional/métodos , Genómica/métodos , Anotación de Secuencia Molecular , Estructura Molecular , Sistemas de Lectura Abierta , Filogenia , Secuencias Repetitivas de Ácidos Nucleicos , Secuenciación Completa del GenomaRESUMEN
Capparis kbangensis Sy & D.V. Hai, a new species from Kbang District, Gia Lai Province, Vietnam, is described and illustrated. The new species is morphologically similar to Capparis versicolor but differs by several characters such as emarginate leaf apex, hairy margin of sepals, smaller fruits, and fewer seeds per fruit. Its ecology and conservation status are provided along with a taxonomic key to the closely allied species.