Phylogenetic placement of Yunnanopilia (Opiliaceae) inferred from molecular and morphological data

Yunnanopilia longistaminea (W.Z. Li) C.Y. Wu & D.Z. Li, which is a controversial species in Opiliaceae, is treated as a variety of Champereia manillana (Blume) Merrill in the Flora of China and morphological analysis has produced conflicting results regarding its affinity to Melientha and Champereia. To determine the systematic position of Yunnanopilia in Opiliaceae, we selected two nuclear regions (internal transcribed spacer [ITS4‐ITS5] and 18S rDNA) and four chloroplast regions (rbcL, matK, psbA‐trnH, and trnS‐trnG) to test the phylogenetics of the family Opiliaceae using maximum likelihood and Bayesian inference analysis. Morphological characteristics were also examined to clarify the similarities and differences among Y. longistaminea and two closely related species. Agonandra was located at the basal position in the family Opiliaceae; in the large clade including other remaining genera, two main clades were clearly identified and correlated with inflorescence morphological characteristics. All samples of Y. longistaminea formed a clade. Yunnanopilia, Melientha, and Champereia were more closely related than other genera of Opiliaceae. Yunnanopilia longistaminea was sister to M. suavis Pierre and was more closely related to M. suavis than to C. manillana. Morphological analysis also showed that differences in the inflorescences and flowers between Y. longistaminea and M. suavis were substantial enough to warrant the retention of Y. longistaminea in its current genus. Thus, we suggest that the monotypic Yunnanopilia be treated as a distinct genus and that the name Y. longistaminea should be adopted.     

Improvement of γ-decalactone production by stimulating the import of ricinoleic acid and suppressing the degradation of γ-decalactone in Saccharomyces cerevisiae

A number of yeast species can transform ricinoleic acid into γ-decalactone, a high-value compound with fruity aroma, through β-oxidation. This study investigated the effect of l-carnitine on γ-decalactone production by Saccharomyces cerevisiae MF013 to increase the β-oxidation rate. Results showed that l-carnitine shortened the biotransformation period by approximately 10?h and increased γ-decalactone production by 19.5%. γ-Caprolactone, γ-octalactone, and γ-dodecalactone were separately added to the medium to prevent γ-decalactone degradation by yeast cells at the end of biotransformation. γ-Octalactone competitively inhibited γ-decalactone from binding to lactonase, resulting in an 11% increase in γ-decalactone production. This research proposed an effective approach to improve the γ-decalactone production rate, shorten the biotransformation period, and suppress the γ-decalactone degradation in S. cerevisiae.     

A genetic analysis of the quality of northern-style Chinese steamed bread

Dissecting the genetic basis for the traits of northern-style Chinese steamed bread (NCSB) is of great significance for wheat quality breeding. Quantitative trait loci (QTLs) for the processing quality of NCSB were studied using a recombinant inbred line (RIL) consisting of 173 lines derived from a “Shannong01–35 × Gaocheng9411” cross. Twenty-four putative additive QTLs were detected on chromosomes 1A, 1B, 1D, 3A, 3B, 4A, 4B, 5B, 6B, and 7B. Of these QTLs, QTex1A.1-27, QHei5B.5-488, and QGum4B.4-17 had the highest contribution and accounted for 9.33, 10.9, and 12.0% of the phenotypic variations, respectively. Several co-located QTLs with additive effects were detected on chromosomes 1A, 1D, 4B, and 5B. Two clusters (RFL_CONTIG2160_524-WSNP_CAP12_C2438_1180601 and EX_C101685_705-RAC875_C27536_611) for height, total score, and texture and for chewiness, gumminess, and hardness were detected on chromosomes 1A and 4B, respectively. Two QTLs for chewiness and hardness (QCh1D-4, QHa1D-4) with additive effects were detected; these alleles could be good targets for improving the processing quality of steamed bread from common wheat (Triticum aestivum L.). In addition, QTLs for wheat flour quality and the associated correlations with NCSB were simultaneously analyzed. Negative correlations were detected between chewiness and the wet/dry gluten content (WGC/DGC) or protein content. Two QTLs (QCh4B.4-17 and QPr4B.4-17) and three QTLs (QCh4B.4-13, QWG4B.4-13, and QDG4B.4-13) clustered in the same chromosomal region. The detected QTL clusters should be further investigated during wheat breeding and could be used by breeders to improve wheat quality and especially the processing quality of NCSB.     

Male germline recombination of a conditional allele by the widely used Dermo1‐cre (Twist2‐cre) transgene

Conditional gene knockout using the Cre/loxP system is instrumental in advancing our understanding of the function of genes in a wide range of disciplines. It is becoming increasingly apparent in the literature that recombination mediated by some Cre transgenes can occur in unexpected tissues. Dermo1‐Cre (Twist2‐Cre) has been widely used to target skeletal lineage cells as well as other mesoderm‐derived cells. Here we report that Dermo1‐Cre exhibits spontaneous male germline recombination activity leading to a Cre‐mediated recombination of a floxed Ptk2 (Protein tyrosine kinase 2, also known as Fak [Focal adhesion kinase]) allele but not a floxed Rb1cc1 (RB1 inducible coiled‐coil 1, also known as Fip200 [FAK‐family Interacting Protein of 200 kDa]) allele at high frequency. This ectopic germline activity of Dermo1‐Cre occurred in all or none manner in a given litter. We demonstrated that the occurrence of germline recombination activity of Dermo1‐Cre transgene can be avoided by using female mice as parental Dermo1‐Cre carriers.     

Investigation of an “alternate water supply system” in enzymatic hydrolysis in the processive endocellulase Cel7A from Rasamsonia emersonii by molecular dynamics simulation

Cel7A from Rasamsonia emersonii is one of the processive endocellulases classified under family 7 glycoside hydrolase. Molecular dynamics simulations were carried out to obtain the optimized sliding and hydrolyzing conformations, in which the reducing ends of sugar chains are located on different sites. Hydrogen bonds are investigated to clarify the interactions between protein and substrate in either conformation. Nine hydrogen bonding interactions are identified in the sliding conformation, and six similar interactions are also found correspondingly in the hydrolyzing conformation. In addition, four strong hydrophobic interactions are also determined. The domain cross‐correlation map analysis shows movement correlation of protein including autocorrelation between residues. The root mean square fluctuations analysis represents the various flexibilities of different fragment in the two conformations. Comparing the two conformations reveals the water‐supply mechanism of selective hydrolysis of cellulose in Cel7A. The mechanism can be described as follow. When the reducing end of substrate slides from the unhydrolyzing site (sliding conformation) to the hydrolyzing site (hydrolyzing conformation), His225 is pushed down and rotated, the rotation leads to the movement of Glu209 with the interstrand hydrogen bonding in β‐sheet. It further makes Asp211 close to the hydrolysis center and provides a water molecule bounding on its carboxyl in the previous unhydrolyzing site. After the hydrolysis takes place and the product is excluded from the enzyme, the Asp211 comes back to its initial position. In summary, Asp211 acts as an elevator to transport outer water molecules into the hydrolysis site for every other glycosidic bond.     

Protective mechanism of FSH against oxidative damage in mouse ovarian granulosa cells by repressing autophagy

Oxidative stress-induced granulosa cell (GCs) death represents a common reason for follicular atresia. Follicle-stimulating hormone (FSH) has been shown to prevent GCs from oxidative injury, although the underlying mechanism remains to be elucidated. Here we first report that the suppression of autophagic cell death via some novel signaling effectors is engaged in FSH-mediated GCs protection against oxidative damage. The decline in GCs viability caused by oxidant injury was remarkably reduced following FSH treatment, along with impaired macroautophagic/autophagic flux under conditions of oxidative stress both in vivo and in vitro. Blocking of autophagy displayed similar levels of suppression in oxidant-induced cell death compared with FSH treatment, but FSH did not further improve survival of GCs pretreated with autophagy inhibitors. Further investigations revealed that activation of the phosphoinositide 3-kinase (PI3K)-AKT-MTOR (mechanistic target of rapamycin [serine/threonine kinase]) signaling pathway was required for FSH-mediated GCs survival from oxidative stress-induced autophagy. Additionally, the FSH-PI3K-AKT axis also downregulated the autophagic response by targeting FOXO1, whereas constitutive activation of FOXO1 in GCs not only abolished the protection from FSH, but also emancipated the autophagic process, from the protein level of MAP1LC3B-II to autophagic gene expression. Furthermore, FSH inhibited the production of acetylated FOXO1 and its interaction with Atg proteins, followed by a decreased level of autophagic cell death upon oxidative stress. Taken together, our findings suggest a new mechanism involving FSH-FOXO1 signaling in defense against oxidative damage to GCs by restraining autophagy, which may be a potential avenue for the clinical treatment of anovulatory disorders.     

Exosomes derived from human amniotic epithelial cells accelerate wound healing and inhibit scar formation

Wound healing is a highly orchestrated physiological process consisting of a complex events, and scarless wound healing is highly desired for the development and application in clinical medicine. Recently, we have demonstrated that human amniotic epithelial cells (hAECs) promoted wound healing and inhibited scar formation through a paracrine mechanism. However, exosomes (Exo) are one of the most important paracrine factors. Whether exosomes derived from human amniotic epithelial cells (hAECs-Exo) have positive effects on scarless wound healing have not been reported yet. In this study, we examined the role of hAECs-Exo on wound healing in a rat model. We found that hAECs, which exhibit characteristics of both embryonic and mesenchymal stem cells, have the potential to differentiate into all three germ layers. hAECs-Exo ranged from 50 to 150 nm in diameter, and positive for exosomal markers CD9, CD63, CD81, Alix, TSG101 and HLA-G. Internalization of hAECs-Exo promoted the migration and proliferation of fibroblasts. Moreover, the deposition of extracellular matrix (ECM) were partly abolished by the treatment of high concentration of hAECs-Exo (100 μg/mL), which may be through stimulating the expression of matrix metalloproteinase-1 (MMP-1). In vivo animal experiments showed that hAECs-Exo improved the skin wound healing with well-organized collagen fibers. Taken together, These findings represent that hAECs-Exo can be used as a novel hope in cell-free therapy for scarless wound healing.