Ester-related volatile compounds reveal the diversity and commonalities of different types of late-ripening peaches.

To recognize the key ester-related volatile compounds, 5 types of peaches including 54 late-ripening peach materials were examined by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry and E-nose. Here, a large number of esters were identified to be released by ripe peach fruits and were mainly characterized by fruity, green, and fatty notes. The variety and content of esters had greatly changed within or between cultivars, indicating that the fruit volatiles were highly differentiated depending on the specific genotypes and cultivation conditions. The ester types showed that fatty acid-derived C6 alcohols and methyl-/ethyl- short-chain alcohol were the main ester precursors, which were more likely to be utilized and well selected by alcohol acyltransferases, whereas the preference of acyl donors was not observed. The common peach type, which exhibited a unique volatile profile, displayed broader diversity and more abundant characteristics in ester-related volatiles than the other four types. A total of 19 key esters were identified as the main components and the content of most esters showed no significant difference among different peach types. Some key esters had even been enriched in nectarines. Moreover, the multiple discriminant analysis revealed a possible relationship between peach types and the domestication of the peach evolution. This study investigated ester-related volatiles released by different types of peach fruits and can be further used to evaluate the peach qualities, providing an important reference for peach breeding and processing.

MdNup62 involved in salt and osmotic stress tolerance in apple.

Abiotic stress of plants has serious consequences on the development of the apple industry. Nuclear pore complexes (NPCs) control nucleoplasmic transport and play an important role in the regulation of plant abiotic stress response. However, the effects of NPCs on apple salt and osmotic stress responses have not been reported yet. In this study, we analyzed the expression and function of NUCLEOPORIN 62 (MdNup62), a component of apple NPC. MdNup62 expression was significantly increased by salt and mannitol (simulated osmotic stress) treatment. The MdNup62-overexpressing (OE) Arabidopsis and tomato lines exhibited significantly reduced salt stress tolerance, and MdNup62-OE Arabidopsis lines exhibited reduced osmotic stress tolerance. We further studied the function of HEAT SHOCK FACTOR A1d (MdHSFA1d), the interacting protein of MdNup62, in salt and osmotic stress tolerance. In contrast to MdNup62, MdHSFA1d-OE Arabidopsis lines showed significantly enhanced tolerance to salt and osmotic stress. Our findings suggest a possible interaction of MdNup62 with MdHSFA1d in the mediation of nuclear and cytoplasmic transport and the regulation of apple salt and osmotic stress tolerance. These results contribute to the understanding of the salt and osmotic stress response mechanism in apple.

Identification and Analysis of Long Non-Coding RNAs Related to UV-B-Induced Anthocyanin Biosynthesis During Blood-Fleshed Peach (Prunus persica) Ripening.

Blood flesh is a key fruit trait in peaches (Prunus persica) and can be attributed to the accumulation of anthocyanins. The roles of long non-coding RNAs (lncRNAs) have been highlighted by multiple studies in regulating fruit ripening, anthocyanin accumulation, and abiotic stress responses in many flowering plants. Such regulatory functions of lncRNAs in Prunus persica, nonetheless, have not been reported. In this research, we sequenced and analyzed the complete transcriptome of C3-20 (a blood-fleshed peach) fruit at four developmental stages. Analyses of the correlated genes and differentially expressed lncRNA target genes helped to forecast lncRNAs' possible functions. The RNA-seq data were generated using high-throughput sequencing. In total, 17,456 putative lncRNAs, including 4,800 intergenic lncRNAs, 2,199 antisense lncRNAs, and 10,439 intronic lncRNAs were discovered, of which 4,871 differentially expressed lncRNAs (DE-lncRNAs) were annotated in the fruit developmental processes. The target genes of these DE-lncRNAs and their regulatory relationship identifying 21,795 cis-regulated and 18,271 trans-regulated targets of the DE-lncRNAs were in a similar way predicted by us. The enriched GO terms for the target genes included anthocyanin biosynthesis. Flavonoid biosynthesis and plant hormone signal transduction were also included in the enriched KEGG pathways. Co-expression network construction demonstrated that the highly expressed genes might co-regulate multiple other genes associated with auxin signal transduction and take effect in equal pathways. We discovered that lncRNAs, including LNC_000987, LNC_000693, LNC_001323, LNC_003610, LNC_001263, and LNC_003380, correlated with fruit that ripened and could take part in ethylene biosynthesis and metabolism and the ABA signaling pathway. Several essential transcription factors, such as ERFs, WRKY70, NAC56, and NAC72, may in a similar way regulate fruit ripening. Three DE-lncRNAs, XLOC_011933, XLOC_001865, and XLOC_042291, are involved in UV-B-induced anthocyanin biosynthesis and positively regulating UVR8 and COP10, were identified and characterized. Our discovery and characterization of XLOC_011933, XLOC_001865, and XLOC_042291 provide a more precise understanding and preliminarily establishes a theoretical framework for UV-B-induced flesh anthocyanin biosynthesis. This phenomenon might encourage more in-depth investigations to study the molecular mechanisms underlying peach flesh coloring.

MdNup54 Interactions With MdHSP70 Involved in Flowering in Apple.

Flowering-related problems in "Fuji" apple have severely restricted the development of China's apple industry. Nuclear pore complexes (NPCs) control nucleoplasmic transport and play an important role in the regulation of plant growth and development. However, the effects of NPCs on apple flowering have not been reported. Here, we analysed the expression and function of MdNup54, a component of apple NPC. MdNup54 expression was the highest in flower buds and maintained during 30-70 days after flowering. MdNup54-overexpressing (OE) Arabidopsis lines displayed significantly earlier flowering than that of the wild type. We further confirmed that MdNup54 interacts with MdHSP70, MdMYB11, and MdKNAT4/6. Consistent with these observations, flowering time of MdHSP70-OE Arabidopsis lines was also significantly earlier. Therefore, our findings suggest a possible interaction of MdNup54 with MdHSP70 to mediate its nuclear and cytoplasmic transport and to regulate apple flowering. The results enhance the understanding of the flowering mechanism in apple and propose a novel strategy to study nucleoporins.

Study of Saponin Components after Biotransformation of Dioscorea nipponica by Endophytic Fungi C39.

This study conducted the solid fermentation process of Dioscorea nipponica using endophytic fungi C39 to determine the changes in the diosgenin concentration. The results revealed that endophytic fungi C39 could effectively biotransform the saponin components in D. nipponica. Moreover, the maximum increase in the diosgenin concentration reached 62.67% in 15 days of solid fermentation. MTT assay results demonstrated that the inhibitory effects of the fermentation drugs on four types of cancer cells (liver cancer cells (HepG2), stomach cancer cells (BGC823), cervical cancer cells (HeLa), and lung cancer cells (A549)) were better than those of the crude drugs obtained from D. nipponica. The chemical composition of the samples obtained before and after the biotransformation of D. nipponica was analyzed by UPLC-Q-TOF-MS. A total of 32 compounds were identified, 21 of which have been reported in Dioscorea saponins and the ChemSpider database and 11 compounds were identified for the first time in D. nipponica. The biotransformation process was inferred based on the variation trend of saponins, which included transformation pathways pertaining to glycolytic metabolism, ring closure reaction, dehydrogenation, and carbonylation. The cumulative findings provide the basis for the rapid qualitative analysis of the saponin components of D. nipponica before and after biotransformation. The 11 metabolites obtained from biotransformation are potential active ingredients obtained from D. nipponica, which can be used to further identify pharmacodynamically active substances.

Analysis of the mechanism of saponin biotransformation in Dioscoreae nipponicae rhizoma by the endophytic fungus Fusarium sp. C39 using whole-genome sequencing.

Fusarium sp. C39 is an endophytic fungus of Dioscorea nipponica Makino. Symbiosis of Fusarium sp. C39 with Rhizoma Dioscoreae Nipponicae (RDN) can significantly increase the content of saponin, which provides a new approach for saponin production and reduces the pressure on natural sources of saponins. However, the underlying mechanism is not clear, limiting its application. In this study, the genome of Fusarium sp. C39 was sequenced, the gene functions were predicted via gene annotation, and the genome was compared to the genomes of four related species. Fusarium sp. C39 is predicted to encode many key enzyme genes involved in saponin synthesis, which could transform the mevalonate, isopentenyl pyrophosphate, and various intermediate compounds present in the RDN extract into saponins. The Fusarium sp. C39 genome contains specific genes that are conducive to its endophytic lifestyle and can provide abundant raw materials for saponin synthesis. Based on the genomic analysis, we proposed the mechanism by which Fusarium sp. C39 generates saponins and provides a theoretical basis for rapid, efficient, low-cost production of saponins.

Advances in steroidal saponins biosynthesis.

MAIN CONCLUSION: This work reviews recent advances in the pathways and key enzymes of steroidal saponins biosynthesis and sets the foundation for the biotechnological production of these useful compounds through transformation of microorganisms. Steroidal saponins, due to their specific chemical structures and active effects, have long been important natural products and that are irreplaceable in hormone production and other pharmaceutical industries. This article comprehensively reviewed the previous and current research progress and summarized the biosynthesis pathways and key biosynthetic enzymes of steroidal saponins that have been discovered in plants and microoganisms. On the basis of the general biosynthetic pathway in plants, it was found that the starting components, intermediates and catalysing enzymes were diverse between plants and microorganisms; however, the functions of their related enzymes tended to be similar. The biosynthesis pathways of steroidal saponins in microorganisms and marine organisms have not been revealed as clearly as those in plants and need further investigation. The elucidation of biosynthetic pathways and key enzymes is essential for understanding the synthetic mechanisms of these compounds and provides researchers with important information to further develop and implement the massive production of steroidal saponins by biotechnological approaches and methodologies.

Immunoregulatory mechanism studies of ginseng leaves on lung cancer based on network pharmacology and molecular docking.

Panax ginseng is one of the oldest and most generally prescribed herbs in Eastern traditional medicine to treat diseases. Several studies had documented that ginseng leaves have anti-oxidative, anti-inflammatory, and anticancer properties similar to those of ginseng root. The aim of this research was to forecast of the molecular mechanism of ginseng leaves on lung cancer by molecular docking and network pharmacology so as to decipher ginseng leaves' entire mechanism. The compounds associated with ginseng leaves were searched by TCMSP. TCMSP and Swiss Target Prediction databases were used to sort out the potential targets of the main chemical components. Targets were collected from OMIM, PharmGKB, TTD, DrugBank and GeneCards which related to immunity and lung cancer. Ginseng leaves exert its lung cancer suppressive function by regulating the several signaling proteins, such as JUN, STAT3, AKT1, TNF, MAPK1, TP53. GO and KEGG analyses indicated that the immunoreaction against lung cancer by ginseng leaves might be related to response to lipopolysaccharide, response to oxidative stress, PI3K-Akt, MAPK and TNF pathway. Molecular docking analysis demonstrated that hydrogen bonding was interaction's core forms. The results of CCK8 test and qRT-PCR showed that ginseng leaves inhibit cell proliferation and regulates AKT1 and P53 expression in A549. The present study clarifies the mechanism of Ginseng leaves against lung cancer and provides evidence to support its clinical use.

Psychological resilience, self-acceptance, perceived social support and their associations with mental health of incarcerated offenders in China.

OBJECTIVES: We aimed to analyze the mental health status of incarcerated offenders in the detention centers and related factors; and introduce psychological resilience, self-acceptance and perceived social support for further analyses, to explore the relationships between them and mental health in these special populations. METHODS: Incarcerated offenders in two detention centers of Guangdong province were recruited. General demographic questionnaire, Self-acceptance Questionnaire (SAQ), Perceived Social Support Scale (PSSS), Connor and Davidson Resilience Scale (CD-RISC) and General Health Questionnaire (GHQ-20) were addressed to the participants for investigation. Pearson correlation analysis, multiple linear regression and Structural Equation Modeling (SEM) were used to identify the associations and related factors. RESULTS: Scores for GHQ-20, CD-RISC, SAQ and PSSS were 7.72 ± 4.49, 57.85 ± 17.30, 40.94 ± 5.40 and 42.99 ± 9.90, respectively. Some socio-demographic factors influenced significantly to the mental health status. After adjusting for socio-demographic factors, the scores of self-acceptance (B = -0.23, P < 0.001), perceived social support (B = -0.10, P < 0.001) and psychological resilience (B = -0.06, P < 0.001) were negatively associated with scores of mental health. Further SEM analyses suggested self-acceptance and resilience had not only mediating effects respectively, but also multiple mediating effects between perceived social support and mental health (r = -0.11, P < 0.01; r = -0.12, P < 0.01; r = -0.04, P < 0.01). CONCLUSION: The levels of mental health, psychological resilience, self-acceptance and perceived social support among incarcerated offenders are generally low, and influenced by socio-demographic factors. In addition, self-acceptance, perceived social support and resilience are protective factors for mental health status of offenders; and self-acceptance and resilience mediate the association between perceived social support and mental health.

A mutation near the active site of S-RNase causes self-compatibility in S-RNase-based self-incompatible plants.

KEY MESSAGE: The structurally simplest amino acid glycine could make contribution to nuclease activity of S-RNase and self-incompatibility in S-RNase-based plants. S-RNase is regarded as inhibitor of self-pollen tube in S-RNase-based self-incompatibility plants. Certain residues like histidine are necessary for RNase activity and self-incompatibility; however, it is unknown whether any other residues contribute to this. Previously, we identified an association between the self-compatible Chinese pear (Pyrus × bretschneideri) cultivar 'Yanzhuang' (YZ) and a mutation causing a residue shift (glycine-to-valine) in the 2nd conserved region (C2) of S21-RNase; however, it was unclear how this nonpolar aliphatic amino acid substitution caused self-compatibility. In this study, we observed that 'YZ' offspring were self-compatible when S21-RNases were all mutated. In vitro pollen tube (S21S21) growth was not completely arrested by the mutated S21-RNase. Residue frequency analysis showed that the glycine residue is highly conserved in diverse S-RNases across many plant species. We therefore generated a mutated petunia SV'-RNase (glycine to valine) and transformed it into S3LS3L petunia. The transformed pistil could not inhibit SV pollen tubes. Three-dimensional protein prediction suggested that the glycine-to-valine mutation alters the spatial structure near the active site, and RNase activity of mutated S-RNase was reducing. Thus, the glycine residue in the C2 is essential for RNase activity, substitution of this residue leads to a failure of self-incompatibility.

Dynamic changes of the main active constituents in valerian rhizome and root (Valeriana amurensis Smir. ex Kom.) during different harvest periods / Cambios dinámicos de los principales constituyentes activos del rizoma y raíz valeriana (Valeriana amurensis Smir. Ex Kom.) durante diferentes períodos de cosecha

Valeriana amurensis Smir. ex Kom. widely distributed in the northeast region of China and some region in Russia and Korea, and its underground parts (roots and rhizomes) being used to cure nervous system diseases such as insomnia. The active components including the essential oil and iridoids of underground parts were investigated in different harvest periods in order to evaluate the quality for the roots and rhizomes of V. amurensis. The content of the essential oil was obtained by hydrodistillation and bornyl acetate in the oil was quantitated by GC-EI. The iridoids, valepotriates were determined by potentiometric titration and the main component, valtrate was quantitated by HPLC-UV. The factors of biomass were considered in the determination of collection period. Statistical analysis of results showed that, the highest content of the essential oil per plant was 22.69 µl in withering period and then 21.58 µl in fruit ripening period, while the highest contents of bornyl acetate, valepotriates and valtrate per plant were 2.82 mg, 31.90 mg and 0.98 mg in fruit ripening period separately. Fruit ripening period was decided as the best harvest period for the content of active constituents and output of drug, and it would provide scientific basis for the artificial cultivation of V. amurensis.

Valeriana amurensis Smir. ex Kom. Se distribuye ampliamente en la región noreste de China y en algunas regiones de Rusia y Corea, y sus partes subterráneas (raíces y rizomas) se utilizan para curar enfermedades del sistema nervioso como el insomnio. Se investigaron los componentes activos, incluidos el aceite esencial y los iridoides de las partes subterráneas de V. amurensis en diferentes períodos de cosecha para evaluar la calidad de las raíces y rizomas. El contenido del aceite esencial se obtuvo mediante hidrodestilación y el acetato de bornilo en el aceite se cuantificó por GC-EI. Los iridoides, valepotriatos se determinaron mediante valoración potenciométrica y el componente principal, el valtrato se cuantificó por HPLC-UV. Los factores de biomasa fueron considerados en la determinación del período de recolección. El análisis estadístico de los resultados mostró que el mayor contenido de aceite esencial por planta fue de 22,69 µl en el período de marchitación y luego de 21,58 µl en el período de maduración de la fruta, mientras que el mayor contenido de acetato de bornilo, valepotriatos y valtrato por planta fue de 2.82 mg, 31.90 mg y 0,98 mg, respectivamente, en el período de maduración de la fruta por separado. Se definió el período de maduración de la fruta como el mejor período de cosecha para el contenido de constituyentes activos y la producción de droga, lo cual proporcionaría una base científica para el cultivo artificial de V. amurensis.

Qualitative Classification of Shear Wave Elastography for Differential Diagnosis Between Benign and Metastatic Axillary Lymph Nodes in Breast Cancer.

Purpose: To examine diagnostic performance of qualitative shear wave elastography (SWE) for evaluation of status of axillary lymph nodes (ALN) in comparison with conventional ultrasonograghy (US) and quantitative SWE parameters. Methods: A total of 118 patients were enrolled, who were all scheduled for breast cancer surgery and core needle biopsy. Conventional US and SWE were performed before biopsy. Based on qualitative evaluation of each ALN, the SWE images were classified into four color patterns: Color Pattern 1: homogeneous; Color Pattern 2: filling defect within lymph node (LN); Color Pattern 3: homogeneous within LN with a localized colored area at the margin; and Color Pattern 4: filling defect within LN with a localized colored area at the margin. The diagnostic performances of the three methods were compared. Results: There were 60 metastatic nodes and 61 benign nodes in the 121 ALNs. Benign ALNs were presented as Color Pattern 1 while metastatic ALNs usually were presented as Color Pattern 2 to 4 (p < 0.05). The AUC of qualitative SWE classification was 0.983, higher than that of quantitative SWE parameters and conventional US (p<0.05). The highest diagnostic performance, with AUC of 0.998, could be achieved if both conventional US and qualitative SWE were applied. Conclusion: The qualitative SWE classification of ALNs proposed in our study exhibited better diagnostic performance than quantitative SWE parameters and conventional US, especially for differentiating metastatic ALNs from benign reactive ALNs. More accurate diagnosis could be reached with this new method and unnecessary biopsy might be avoided in the meantime.

Screening for differentially expressed genes in endophytic fungus strain 39 during co-culture with herbal extract of its host Dioscorea nipponica Makino.

Strain 39 is an endophytic fungus which was isolated from Dioscorea nipponica Makino (DNM). After Strain 39 co-cultured with ethanol extract of DNM rhizomes for several days, the content of saponins in this culture mixture would be obviously increased. To analyze the mechanism of this microbial transformation, we used the differential display reverse transcription polymerase chain reaction (DDRT-PCR) method to compare the transcriptomes between Strain 39 cultured in normal PD medium and in PD medium which added ethanol extract of DNM rhizomes. We amplified 29 DDRT-PCR bands using 12 primer combinations of three anchored primers and five random primers, and six bands were re-amplified. Analysis of real-time PCR and sequence alignment showed that three clones were up-regulated in sample group: squalene epoxidase, squalene synthase, and catalase, one clone was expressed only in sample group. The possible roles and origins of the above genes were discussed, and the molecular mechanism of Strain 39 biotransformation was speculated. This study is the first report of the molecular biotransformation mechanism of saponins production by endophytic fungus of DNM.

A caryophyllane-type sesquiterpene, caryophyllenol A from Valeriana amurensis.

A new caryophyllane derivative, caryophyllenol A, and a new germacrane derivative, isovolvalerenal D, together with 11 known sesquiterpenoids, were isolated from a petroleum ether partition of the roots and rhizomes of Valeriana amurensis. Structure elucidation of caryophyllenol A and isovolvalerenal D was accomplished on the basis of various spectroscopic techniques including HRESIMS and 2D NMR analyses. The structure of caryophyllenol A was further confirmed by X-ray crystallography and using quantum-chemical ECD calculation adopting TDDFT method. Caryophyllenol A and other eight sesquiterpenoids were evaluated for sedative activity with the model of Drosophila melanogaster, and eight of them showed the effect of prolonging the total sleeping time (TST) of D. melanogaster, displaying significant sedative action.

Genetic features of the spontaneous self-compatible mutant, 'Jin Zhui' (Pyrus bretschneideri Rehd.).

'Jin Zhui' is a spontaneous self-compatible mutant of 'Ya Li' (Pyrus bretschneideri Rehd. S21S34 ), the latter displaying a typical S-RNase-based gametophytic self-incompatibility (GSI). The pollen-part mutation (PPM) of 'Jin Zhui' might be due to a natural mutation in the pollen-S gene (S34 haplotype). However, the molecular mechanisms behind these phenotypic changes are still unclear. In this study, we identified five SLF (S-Locus F-box) genes in 'Ya Li', while no nucleotide differences were found in the SLF genes of 'Jin Zhui'. Further genetic analysis by S-RNase PCR-typing of selfed progeny of 'Jin Zhui' and 'Ya Li' × 'Jin Zhui' progeny showed three progeny classes (S21S21 , S21S34 and S34S34 ) as opposed to the two classes reported previously (S21S34 and S34S34 ), indicating that the pollen gametes of 'Jin Zhui', bearing either the S21 - or S34 -haplotype, were able to overcome self-incompatibility (SI) barriers. Moreover, no evidence of pollen-S duplication was found. These findings support the hypothesis that loss of function of S-locus unlinked PPM expressed in pollen leads to SI breakdown in 'Jin Zhui', rather than natural mutation in the pollen-S gene (S34 haplotype). Furthermore, abnormal meiosis was observed in a number of pollen mother cells (PMCs) in 'Jin Zhui', but not in 'Ya Li'. These and other interesting findings are discussed.

Identification of a canonical SCF(SLF) complex involved in S-RNase-based self-incompatibility of Pyrus (Rosaceae).

S-RNase-based self-incompatibility (SI) is an intraspecific reproductive barrier to prevent self-fertilization found in many species of the Solanaceae, Plantaginaceae and Rosaceae. In this system, S-RNase and SLF/SFB (S-locus F-box) genes have been shown to control the pistil and pollen SI specificity, respectively. Recent studies have shown that the SLF functions as a substrate receptor of a SCF (Skp1/Cullin1/F-box)-type E3 ubiquitin ligase complex to target S-RNases in Solanaceae and Plantaginaceae, but its role in Rosaceae remains largely undefined. Here we report the identification of two pollen-specific SLF-interacting Skp1-like (SSK) proteins, PbSSK1 and PbSSK2, in Pyrus bretschneideri from the tribe Pyreae of Rosaceae. Both yeast two-hybrid and pull-down assays demonstrated that they could connect PbSLFs to PbCUL1 to form a putative canonical SCF(SLF) (SSK/CUL1/SLF) complex in Pyrus. Furthermore, pull-down assays showed that the SSK proteins could bind SLF and CUL1 in a cross-species manner between Pyrus and Petunia. Additionally, phylogenetic analysis revealed that the SSK-like proteins from Solanaceae, Plantaginaceae and Rosaceae form a monoclade group, hinting their shared evolutionary origin. Taken together, with the recent identification of a canonical SCF(SFB) complex in Prunus of the tribe Amygdaleae of Rosaceae, our results show that a conserved canonical SCF(SLF/SFB) complex is present in Solanaceae, Plantaginaceae and Rosaceae, implying that S-RNase-based self-incompatibility shares a similar molecular and biochemical mechanism.

[Pharmacological effects of volatile oil of Valeriana amurensis on CNS].

OBJECTIVE: To study the pharmacological effects of volatile oil of Valeriana amurensis on central nervous system. METHODS: The pharmacological effects of volatile oil of Valeriana amurensis on the autonomic activities of mice, the sleeping synergistic action of mice with pentoharbital sodium at subthreshold and hypnotic dosages, the sleep phases of rats, the writhing response of mice caused by acetic acid and the convulsion of mice induced by thio-semicarbazide were investigated. RESULTS: The autonomic activities of mice were significantly inhibited by the volatile oil of Valeriana amurensis. The rate of falling sleep and the extension of sleeping time of mice were significantly increased by the synergic action of pentobarbital sodium with the volatile oil. The sleep phases of SWS2 and REMS of rats were obviously extended by the volatile oil of Valeriana amurensis. In addition, the frequency of writhing response of mice caused by acetic acid was reduced, and the convulsion of mice induced by thio-semicarbazide was antagonized with the administration of the volatile oil. CONCLUSION: The volatile oil of Valeriana amurensis has significantly sedative analgesic and anti-hyperspasmia effects.