We report isolation and characterization of a fragile culm mutant fc116 that displays reduced mechanical strength caused by decreased cellulose content and altered cell wall structure in rice. Map-based cloning revealed that fc116 was a base substitution mutant (G to A) in a putative beta-1,6-N-acetylglucosaminyltransferase (C2GnT) gene (LOC_Os05g07790, allelic to BC10). This mutation resulted in one amino acid missing within a newly-identified protein motif “R, RXG, RA.” The FC116/BC10 gene was lowly but ubiquitously expressed in the all tissues examined across the whole life cycle of rice, and slightly down-regulated during secondary growth. This mutant also exhibited a significant increase in the content of hemicelluloses and lignins, as well as the content of pentoses (xylose and arabinose). But the content of hexoses (glucose, mannose, and galactose) was decreased in both cellulosic and non-cellulosic (pectins and hemicelluloses) fractions of the mutant. Transcriptomic analysis indicated that the typical genes in the fc116 mutant were up-regulated corresponding to xylan biosynthesis, as well as lignin biosynthesis including p-hydroxyphenyl (H), syringyl (S), and guaiacyl (G). Our results indicate that FC116 has universal function in regulation of the cell wall polymers in rice.
Antimicrobial peptides (AMPs) are effective against different plant pathogens and newly considered as part of plant defense systems. From prokaryotes to eukaryotes, AMPs can exist in all forms of life. SM-985 is a cationic AMP (CAMP) isolated from the cDNA library of Mexican teosinte (Zea mays ssp. mexicana). A computational prediction server running with different algorithms was used to screen the teosinte cDNA library for AMPs, and the SM-985 peptide was predicted as an AMP with high probability prediction values. SM-985 is an arginine-rich peptide and composed of 21 amino acids (MW: 2671.06 Da). The physicochemical properties of SM-985 are very promising as an AMP, including the net charge (+8), hydrophobicity ratio of 23%, Boman index of 5.19 kcal/mol, and isoelectric point of 12.95. The SM-985 peptide has amphipathic α-helix conformations. The antimicrobial activity of SM-985 was confirmed against six bacterial plant pathogens, and the MIC of SM-985 against Gram-positive indicators was 8 µM, while the MIC of SM-985 against Gram-negative indicators was 4 µM. The SM-985 interacting with the bacterial membrane and this interaction were examined by treatment of the bacterial indicators with FITC-SM-985 peptide, which showed a high binding affinity of SM-985 to the bacterial membrane (whether Gram-positive or Gram-negative). Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images of the treated bacteria with SM-985 demonstrated cell membrane damage and cell lysis. In vivo antimicrobial activity was examined, and SM-985 prevented leaf spot disease infection caused by Pst DC3000 on Solanum lycopersicum. Moreover, SM-985 showed sensitivity to calcium chloride salt, which is a common feature of CAMPs.
Thyme essential oil‐loaded microcapsules (TMS) were fabricated using natural polysaccharide chitosan (CS) and sodium alginate (SA) as the shell material via the method of layer‐by‐layer (LBL) assembly. The accumulated release rates of thyme oil and microcapsules at 4 °C were 42.50% and 10.16%, respectively. After heating at 100 °C for 5 hr, the release rate of the 0, 2, 4, 6 layers assembled microcapsules were 100%, 48.84%, 28.38%, 19.3%, severally. Microcapsules also had good pH sensitivity in the range of 4 to 10. Antimicrobial function studies showed that the microcapsules are more effective than thyme oil for three tested microorganisms. When the temperature rose from 37 °C to 121 °C, the antibacterial zone of thyme oil gradually decreased from 18.5 ± 0.6 mm to 12.3 ± 0.6 mm, although inhibition rate of microcapsules increased from 87.97% to 99.75%. The antibacterial effect of thyme oil declined with the increase of pH, in terms of microcapsules, the efficiency was better under acidic or alkaline conditions. The thyme oil microcapsules can suppress the growth of Staphylococcus aureus in milk and prolong its shelf life. It was determined that this microcapsule could be a potential alternative as a natural antimicrobial agent in food and pharmaceutical industries.
Practical Application
This work provided release performance and mechanism of layer‐by‐layer (LBL) thyme oil microcapsule under different conditions, and further studies showed its antibacterial ability to explore how herb essential oils can be potentially applied in food packaging and antibacterial areas.
Legumes in the inverted repeat-lacking clade (IRLC) each produce a unique set of nodule-specific cysteine-rich (NCR) peptides, which act in concert to determine the terminal differentiation of nitrogen-fixing bacteroid. IRLC legumes differ greatly in NCR number and sequence diversity. This raises a significant question how bacteroid differentiation is collectively controlled by the specific NCR repertoire of an IRLC legume. Astragalus sinicus is an IRLC legume that forms indeterminate nodules with its microsymbiont Mesorhizobium huakuii 7653R. Here, we performed transcriptome analysis of root and nodule samples at 3, 7, 14, 28 days post inoculation with M. huakuii 7653R and its isogenic ∆bacA mutant. BacA is a broad-specificity peptide transporter required for the host-derived NCRs to target rhizobial cells. A total of 167 NCRs were identified in the RNA transcripts. Comparative sequence and electrochemical analysis revealed that A. sinicus NCRs (AsNCRs) are dominated by a unique cationic group (termed subgroup C), whose mature portion is relatively long (>60 amino acids) and phylogenetically distinct and possessing six highly conserved cysteine residues. Subsequent functional characterization showed that a 7653R variant harboring AsNCR03 (a representative of subgroup C AsNCR) displayed significant growth inhibition in laboratory media, and formed ineffective white nodules on A. sinicus with irregular symbiosomes. Finally, bacterial two-hybrid analysis led to the identification of GroEL1 and GroEL3 as the molecular targets of AsNCR067 and AsNCR076. Together, our data contribute to a systematic understanding of the NCR repertoire associated with the A. sinicus and M. huakuii symbiosis.
The aim of this study was to produce and characterize microcapsules of thyme oil and finally appraise the extent of stability improvement. The optimum process conditions obtained from orthogonal tests were as follows: ratio of core material to wall 0.5, temperature 40 °C, pH value 3.0 and time 20 min, where the practical encapsulation efficiency was 85.17±1.35%. The microcapsules belong to the nanometric range as the average particle diameter was 531.17±77.12 nm. The results from structural analysis indicated that no significant chemical bond occurred during the encapsulation process and the microcapsules remained stable when the encapsulation was conducted at a temperature below 53.1 °C. Especially, the retention rate of thyme oil in microcapsules remained 39.21% at 4 °C, 36.99% at 25 °C and 33.80% at 40 °C after 30 d of storage. Moreover, protection from light exposure presented a positive impact on the storage stability of thyme oil microcapsules.
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