Search

Browning presents a significant challenge in the in vitro culture of economically important woody plants, primarily due to high levels of lignification and the accumulation of secondary metabolites. This phenomenon hampers the development of efficient regeneration and genetic transformation systems across diverse species. This review examines the internal and external factors contributing to browning, including genetic attributes, tree genotypes, physiological state of explants, explant surface sterilization, medium composition, and overall culture conditions. It explores the underlying mechanisms of browning, particularly enzymatic browning caused by the oxidation of phenolic compounds, and highlights the crucial role of redox pathways and phenolic metabolism. Conventional methods for assessing browning, such as sensory evaluation by researchers and the examination of paraffin sections stained with toluidine blue, are commonly used but introduce significant delays and potential biases. The review emphasizes the importance of accurate and timely browning assessment methods, notably the use of Fluorescein diacetate (FDA) staining, as a reliable and quantitative measure of cell viability to better evaluate browning intensity and progression. Additionally, this review explores the potential manipulation of key genes in the phenylpropanoid pathway to lower phenolic biosynthesis. Advanced strategies, such as regenerative gene manipulation and natural product encapsulation, are also discussed for their potential to improve regeneration outcomes. By integrating recent advancements in molecular biology and tissue culture techniques, this review offers novel insights and potential solutions for mitigating browning, thereby enhancing the regeneration capacities of woody plants. This comprehensive approach addresses the mechanistic bases of browning and underscores the importance of optimizing cultural practices and genetic strategies to overcome this challenge.

Abstract It is widely known that some soils have strong levels of disease suppression and prevent the establishment of pathogens in the rhizosphere of plants. However, what soils are better suppressing disease, and how management can help us to boost disease suppression remain unclear. Here, we used field, greenhouse and laboratory experiments to investigate the effect of management (monocropping and rotation) on the capacity of rhizosphere microbiomes in suppressing peanut root rot disease. Compared with crop rotations, monocropping resulted in microbial assemblies that were less effective in suppressing root rot diseases. Further, the depletion of key rhizosphere taxa in monocropping, which were at a disadvantage in the competition for limited exudates resources, reduced capacity to protect plants against pathogen invasion. However, the supplementation of depleted strains restored rhizosphere resistance to pathogen. Taken together, our findings highlight the role of native soil microbes in fighting disease and supporting plant health, and indicate the potential of using microbial inocula to regenerate the natural capacity of soil to fight disease.

Ginkgo biloba, usually referred to as a “living fossil,” is widely planted in many countries because of its medicinal value and beautiful appearance. Owing to ginkgo’s high resistance to drought stress, ginkgo seedlings can even survive withholding water for several days without exhibiting leaf wilting and desiccation. To assess the physiological and transcriptomic mechanisms involved in the drought stress and re-watering responses of Ginkgo biloba, ginkgo seedlings were subjected to drought treatment for 15 d (D_15 d) and 22 d (D_22 d) until they had severely wilted, followed by re-watering for 3 d (D_Re3 d) to restore normal growth. Variations in physiological characteristics (relative water content, malondialdehyde (MDA) content, stomatal aperture, and antioxidant enzyme activity) during drought and re-watering were assessed. In total, 1692, 2031, and 1038 differentially expressed genes (DEGs) were upregulated, while 1691, 2820, and 1910 were downregulated in D_15 d, D_22 d, and D_Re3 d, respectively, relative to the control. Three pathways, namely, plant hormone signal transduction, plant–pathogen interaction, and the plant MAPK signaling pathway, were enriched during drought stress and re-watering. The DEGs involved in plant hormone signal transduction pathways (those of IAA, CTK, GA, ABA, ETH, BR, SA, and JA) and the major differentially expressed transcription factors (TFs; MYB, bHLH, AP2/ERF, NAC, WRKY, and bZIP) were identified. Quantitative real-time PCR revealed six TFs as positive or negative regulators of drought stress response. These phenotype-related physiological characteristics, DEGs, pathways, and TFs provide valuable insights into the drought stress and re-watering responses in G. biloba.

In exploring the male–female differentiation and differences in the seedling stage of the F1 generation of ginkgo hybrids, an early selection test for the cultivation and research of leafy or medicinal ginkgo industry was conducted, which may provide a basis for boosting the precision of the ginkgo industry. The hybrid F1 generation obtained from the cross-mating was used as material for the determination of growth and development, as well as of the physiology and biochemistry of the monocots, and the male and female differential genes were obtained based on the data of SNPs obtained from the GBS sequencing of the hybrid progeny. In the seedling stage of ginkgo hybrid offspring, male plants had a significantly higher nutrient growth capacity than female plants, while the total flavonoid and terpene lactone contents in female plants were higher than those of the male plants. This result can provide a corresponding theoretical basis for the use of ginkgo germplasm resources, which can make full use of the male and female differences in the seedling stage and maximize the benefits of early sex identification.

Abstract Nitrate is a primary nitrogen source for plant growth, and previous studies have indicated a correlation between nitrogen and browning. Nitrate transporters (NRTs) are crucial in nitrate allocation. Here, we utilized a genome-wide approach to identify and analyze the expression pattern of 74 potential GbNRTs under nitrate treatments during calluses browning in Ginkgo, including 68 NITRATE TRANSPORTER 1 (NRT1)/PEPTIDE TRANSPORTER (PTR) (NPF), 4 NRT2 and 2 NRT3. Conserved domains, motifs, phylogeny, and cis-acting elements (CREs) were analyzed to demonstrate the evolutionary conservation and functional diversity of GbNRTs. Our analysis showed that the NPF family was divided into eight branches, with the GbNPF2 and GbNPF6 subfamilies split into three groups. Each GbNRT contained 108–214 CREs of 19–36 types, especially with binding sites of auxin and transcription factors v-myb avian myeloblastosis viral oncogene homolog (MYB) and basic helix-loop-helix (bHLH). The E1X1X2E2R motif had significant variations in GbNPFs, indicating changes in the potential dynamic proton transporting ability. The expression profiles of GbNRTs indicated that they may function in regulating nitrate uptake and modulating the signaling of auxin and polyphenols biosynthesis, thereby affecting browning in Ginkgo callus induction. These findings provide a better understanding of the role of NRTs during NO3 − uptake and utilization in vitro culture, which is crucial to prevent browning and develop an efficient regeneration and suspension production system in Ginkgo.

Quercus gilva is a precious timber tree species in middle subtropical China. In order to investigate the phenotypic diversity, phenotypic variation pattern and the major geographic and climatic factors influencing phenotypic variation of Q. gilva populations, 15 growth and leaf traits of 115 individuals from 14 natural populations were measured, and then the phenotypic diversity, variation pattern and the correlation between phenotypic variation and geographic and climatic factors were studied using nested analysis of variance, phenotypic differentiation coefficient, diversity index, correlation analysis, principal component analysis and cluster analysis. The results were as follows: (1) The average variation coefficient and the average Shannon-Wiener index of 15 phenotypic traits were 35.070% and 1.998, respectively. Variation coefficient and Shannon-Wiener index of 14 populations ranged from 14.94% (Dongkou) to 35.56% (Longshan) and from 1.127 (Songyang) to 1.980 (Changning), respectively. These results indicated that Q. gilva showed a relatively high level of phenotypic diversity. (2) Significant differences in 15 phenotypic traits were found among and within populations (P

Soil salinization significantly impacts the ecological environment and agricultural production, posing a threat to plant growth. Currently, there are over 400 varieties of Bougainvillea with horticultural value internationally. However, research on the differences in salt tolerance among Bougainvillea varieties is still insufficient. Therefore, this study aims to investigate the physiological responses and tolerance differences of various Bougainvillea varieties under different concentrations of salt stress, reveal the effects of salt stress on their growth and physiology, and study the adaptation mechanisms of these varieties related to salt stress. The experimental materials consisted of five varieties of Bougainvillea. Based on the actual salinity concentrations in natural saline-alkali soils, we used a pot-controlled salt method for the experiment, with four treatment concentrations set: 0.0% (w/v) (CK), 0.2% (w/v), 0.4% (w/v), and 0.6% (w/v). After the Bougainvillea plants grew stably, salt stress was applied and the growth, physiology, and salt tolerance of the one-year-old plants were systematically measured and assessed. The key findings were as follows: Salt stress inhibited the growth and biomass of the five varieties of Bougainvillea; the ‘Dayezi’ variety showed severe salt damage, while the ‘Shuihong’ variety exhibited minimal response. As the salt concentration and duration of salt stress increase, the trends of the changes in antioxidant enzyme activity and osmotic regulation systems in the leaves of the five Bougainvillea species differ. Membrane permeability and the production of membrane oxidative products showed an upward trend with stress severity. The salt tolerance of the five varieties of Bougainvillea was comprehensively evaluated through principal component analysis. It was found that the ‘Shuihong’ variety exhibited the highest salt tolerance, followed by the ‘Lvyehuanghua’, ‘Xiaoyezi’, ‘Tazi’, and ‘Dayezi’ varieties. Therefore, Bougainvillea ‘Shuihong’, ‘Lvyehuanghua’, and ‘Xiaoyezi’ are recommended for extensive cultivation in saline-alkali areas. The investigation focuses primarily on how Bougainvillea varieties respond to salt stress from the perspectives of growth and physiological levels. Future research could explore the molecular mechanisms behind the responses to and tolerance of different Bougainvillea varieties as to salt stress, providing a more comprehensive understanding and basis for practical applications.

Arrow bamboo (Fargesia qinlingensis) is endemic to the Qinling Mountains and has remarkable adaptive resilience to changing climates. However, its complete mitogenome remains unknown. Using the Illumina and PacBio HiFi sequencing platforms, we found that the mitogenome assembly of the F. qinlingensis has a multi-branched skeleton comprising three linear molecules (M1, M2, and M3), with a length of 442,368 bp and a GC content of 44.05%. Thirty-five unique PCGs were identified in the complete mitogenome, including twenty-four core structural genes, eleven noncore structural genes, three rRNAs, and sixteen tRNAs. The GCU for alanine and CAA for glutamine represented the most significant frequency (RSCU = 1.55) in the codon usage preference. A total of 51, 28, and 14 SSRs were determined on M1, M2, and M3, respectively. The mitogenome contained 149 pairs of dispersed repeats with lengths greater than 30 bp, the most abundant of which were 82 forward and 67 palindromic repeats. A long repeat sequence (14,342 bp) was characterized in mediating mitogenome recombination. DNA transfer analyses suggested that 44 MTPTs (30,943 bp, 6.99%) originated from the plastome. Among the 482 potential C-U/T RNA-editing sites predicted in 35 PCGs, ccmFn (38 times) and ccmC (36 times) shoed the highest frequency. Collinearity and phylogenetic trees revealed the close relationship between F. qinlingensis and Bambusa oldhamii. The primary features of the mitogenome of F. qinlingensis will help decipher the functional mitochondrial traits related to growth performance and climate resilience. Moreover, our findings provide insights into the evolution, environmental adaptation, and sustainable use of subalpine bamboo resources in the Qinling Mountains.

Abstract Introduction Endoscopic ultrasound (EUS) may play a role in evaluating treatment response after definitive chemoradiation therapy (dCRT) for esophageal squamous cell carcinoma (ESCC). This study explored the prognostic markers of EUS with biopsies and developed two nomograms for survival prediction. Methods A total of 821 patients newly diagnosed with ESCC between January 2015 and December 2019 were reviewed. We investigated the prognostic value of the changes in tumor imaging characteristics and histopathological markers by an interim response evaluation, including presence of stenosis, ulceration, tumor length, tumor thickness, lumen involvement, and tumor remission. Independent prognostic factors of progression-free survival (PFS) and overall survival (OS) were determined using Cox regression analysis and further selected to build two nomogram models for survival prediction. The receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA) were used to respectively assess its discriminatory capacity, predictive accuracy, and clinical usefulness. Results A total of 155 patients were enrolled in this study and divided into the training (109 cases) and testing (46 cases) cohorts. Tumor length, residual tumor thickness, reduction in tumor thickness, lumen involvement, and excellent remission (ER) of spatial luminal involvement in ESCC (ER/SLI) differed significantly between responders and non-responders. For patients undergoing dCRT, tumor stage (P = 0.001, 0.002), tumor length (P = 0.013, 0.008), > 0.36 reduction in tumor thickness (P = 0.004, 0.004) and ER/SLI (P = 0.041, 0.031) were independent prognostic markers for both PFS and OS. Time-dependent ROC curves, calibration curves, and DCA indicated that the predicted survival rates of our two established nomogram models were highly accurate. Conclusion Our nomogram showed high accuracy in predicting PFS and OS for ESCC after dCRT. External validation and complementation of other biomarkers are needed in further studies.

Ginkgo is an economic tree species with high medicinal value, and flavonols are its main medicinal components. This research was conducted to investigate the molecular mechanism underlying the influence of Ultraviolet A (UVA) treatment on the synthesis of ginkgo flavonols with the aim of increasing their content. Ginkgo full-sib hybrid offspring were used as test materials. The phenylalanine ammonialyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumarate: CoA ligase (4CL) enzyme activities, as well as flavonol contents, were measured under the same intensity of white light (300 μmol·m−2·s−1) with the addition of 20, 40, and 60 μmol·m−2·s−1 UVA separately after 20 days of treatment. The control check (CK) and treatment with the highest flavonol content were chosen for transcriptome sequencing analysis. The results showed that the PAL, C4H, and 4CL enzyme activities, as well as the flavonol and totalflavonol glycoside contents, of ginkgo hybrid progeny differed significantly under different UVA treatments. They showed a tendency to increase and then decrease, reaching a maximum value under UVA-4 (40 μmol·m−2·s−1 ultraviolet UVA light intensity) treatment. Ribonucleic acid (RNA) sequencing revealed the presence of 4165 genes with differential expression, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the metabolic pathways commonly enriched across all four comparison groups included ‘phenylpropanoid biosynthesis’, while the pathways commonly enriched in green-leaf ginkgo UVA-4 treatment (TL), yellow-leaf ginkgo mutant CK treatment (CKY), and green-leaf ginkgo CK treatment (CKL) were related to ‘flavonoid biosynthesis’. Treatment with UVA light led to the increased expression of PAL and 4CL enzymes in the phenylpropanoid biosynthesis pathway, as well as increased expression of chalcone synthase (CHS), Flavanone 3-hydroxylase (F3H), and flavonol synthase (FLS) enzymes in the flavonoid biosynthesis pathway, thereby promoting the synthesis of ginkgo flavonols. In summary, the use of 40 μmol·m−2·s−1 UVA treatment for 20 days significantly increased the flavonol content and the expression of related enzyme genes in ginkgo hybrid offspring, enhancing ginkgo flavonoids and increasing the medicinal value of ginkgo.

The Plateau Hongliu (Myricaria elegans Royle.) is a woody shrub halophyte that thrives in arid areas of western Tibet, in the Himalayan Mountains. It is acclaimed as superior in saline stress acclimation and as a critical pharmaceutical resource of the Tibetan traditional herb. Nevertheless, the mitogenome in the genus Myricaria remains unknown. Here, using the Illumina and PacBio sequencing assays, the first complete mitogenome of the M. elegans revealed a multi-branched skeleton with a total length of 416,354 bp and GC content of 44.33%, comprising two circular molecules (M1 and 2). The complete mitogenome annotates 31 unique protein-encoding genes (PEGs), fifteen tRNAs, and three rRNA genes. The UAA exhibits the most prominent codon usage preference as a termination, followed by UUA codons for leucine. The mitogenome contains 99 simple sequence repeats and 353 pairs of dispersed repeats, displaying the most frequent in palindromic repeats. Gene transfer analyses identified 8438 bp of 18 homologous fragments from the plastome, accounting for 2.03% of the total length. Using the PREP suite, 350 C-U RNA editing sites were predicted, of which nad4 and ccmB were on the top frequency. Syntenic and phylogenetic analyses suggested weakly conserved patterns of M. elegans in Caryophyllales owing to the genome rearrangement. In summary, the deciphered unique features and complexities of the mitogenome in M. elegans provide novel insights into understanding the evolution and biological conservation underlying climate resilience in halophytes.

Establishing plant regeneration systems and efficient genetic transformation techniques plays a crucial role in plant functional genomics research and the development of new crop varieties. The inefficient methods of transformation and regeneration of recalcitrant species and the genetic dependence of the transformation process remain major obstacles. With the advancement of plant meristematic tissues and somatic embryogenesis research, several key regulatory genes, collectively known as developmental regulators, have been identified. In the field of plant genetic transformation, the application of developmental regulators has recently garnered significant interest. These regulators play important roles in plant growth and development, and when applied in plant genetic transformation, they can effectively enhance the induction and regeneration capabilities of plant meristematic tissues, thus providing important opportunities for improving genetic transformation efficiency. This review focuses on the introduction of several commonly used developmental regulators. By gaining an in-depth understanding of and applying these developmental regulators, it is possible to further enhance the efficiency and success rate of plant genetic transformation, providing strong support for plant breeding and genetic engineering research.

Hippophae gyantsensis Lian is a pioneering tree species in Chinese forestry ecological engineering, known for its robust stress tolerance, water retention capacity, and soil improvement qualities. However, the lack of rapid nursery technology has been a significant impediment to the development of the H. gyantsensis industry. In the present study, we have successfully established a tissue culture regeneration system for H. gyantsensis. The most effective methods for seed disinfection, ensuring sterility in seedlings, were found to be 75% alcohol disinfection for 40 s and 10% sodium hypochlorite disinfection for 10 min. The best media tested for callus induction in cotyledons and hypocotyls of sterile seedlings were 1/3 MS + 0.5 mg/L KT + 0.75 mg/L NAA and 1/3 MS + 0.3 mg/L 6-BA + 1.5 mg/L IBA, respectively. As the explants, cotyledons yielded larger calli with a greater size and differentiation ability than hypocotyls. For the induction of indeterminate shoots and proliferation, the most suitable media were 1/3 MS + 0.5 mg/L IAA + 0.75 mg/L 6-BA and 1/3 MS + 1.0 mg/L 6-BA + 0.05 mg/L IBA + 0.1 mg/L KT, respectively. Lastly, the best worked rooting formulation was 1/4 MS + 0.3 mg/L IBA. This study marks a significant milestone in the establishment of a systematic tissue culture regeneration system for H. gyantsensis, which will facilitate the industrial rapid propagation of high-quality seedlings and provide the foundation for improvement through genetic transformation.

Flavonoids are abundant secondary metabolites in Ginkgo biloba and have a wide range of medicinal values. Salicylic acid (SA) can induce flavonoid accumulation in plants, but the detailed regulatory mechanism remains unclear. Here, we established the optimal media for ginkgo callus induction and subculture, and found that exogenous SA greatly increased the content of flavonol, including quercetin, kaempferol, isorhamnetin. Transcriptome changes in SA-treated calli showed that most structural genes involved in flavonoid biosynthesis were upregulated. Particularly, overexpression of GbF3′H in ginkgo calli significantly increased the content of flavonol, suggesting the vital role of GbF3′H in flavonoid biosynthesis. We further identified that a R2R3-MYB, GbMYB36, were significantly upregulated in SA treated calli. Transient overexpression and a LUC assay indicate that GbMYB36 act as an activator, and improve flavonoid biosynthesis through regulating the expression of GbF3′H. Our findings provide insight into the molecular basis of SA-induced flavonoid biosynthesis in ginkgo.

Alcoholic liver disease (ALD) is primarily caused by long-term excessive alcohol consumption. Cyanidin-3-O-glucoside (C3G) is a widely occurring natural anthocyanin with multiple biological activities. This study aims to investigate the effects of C3G isolated from black rice on ALD and explore the potential mechanism. C57BL/6J mice (male) were fed with standard diet (CON) and Lieber-DeCarli liquid-fed (Eth) or supplemented with a 100 mg/kg/d C3G Diet (Eth-C3G), respectively. Our results showed that C3G could effectively ameliorate the pathological structure and liver function, and also inhibited the accumulation of liver lipids. C3G supplementation could partially alleviate the injury of intestinal barrier in the alcohol-induced mice. C3G supplementation could increase the abundance of Norank_f_Muribaculaceae, meanwhile, the abundances of Bacteroides, Blautia, Collinsella, Escherichia-Shigella, Enterococcus, Prevotella, [Ruminococcus]_gnavus_group, Methylobacterium-Methylorubrum, Romboutsia, Streptococcus, Bilophila, were decreased. Spearman’s correlation analysis showed that 12 distinct genera were correlated with blood lipid levels. Non-targeted metabolic analyses of cecal contents showed that C3G supplementation could affect the composition of intestinal metabolites, particularly bile acids. In conclusion, C3G can attenuate alcohol-induced liver injury by modulating the gut microbiota and metabolites, suggesting its potential as a functional food ingredient against alcoholic liver disease.

The WUSCHEL-related homeobox (WOX) gene family plays a crucial role in regulating embryonic development, organ formation, and stress resistance. Yellowhorn (Xanthoceras sorbifolia Bunge), a drought-resistant tree known for its oil production, lacks sufficient information regarding the WOX gene family. To understand the evolutionary mechanisms and potential functions of this gene family in yellowhorn, we conducted a comprehensive investigation on its expression patterns and evolutionary characteristics. Our analysis revealed the presence of nine XsWOX genes in the yellowhorn genome, which could be categorized into three distinct clades through a phylogenetic analysis. A chromosomal localization analysis indicated that these nine XsWOX genes were situated on six out of the fifteen chromosomes. An intra-species collinear analysis revealed only one pair of tandem duplicated genes within the XsWOX family. The promoter regions of the XsWOX family were found to contain responsive cis-acting elements associated with plant growth and development, stress responses, and hormone signaling. Moreover, an analysis of the gene expression profiles in different developmental stages of callus revealed significant expressions of XsWOX1, XsWOX4, and XsWOX5 in embryogenic callus and somatic embryo formation, suggesting that they have special roles in regulating yellowhorn’s somatic embryogenesis. Furthermore, the expression level of XsWOX5 indicated its potential involvement not only in organ formation but also in responding to low temperature, salt, and saline-alkali stresses. Overall, our findings lay a solid foundation for future in-depth studies on the functionality and evolution of XsWOX genes in yellowhorn.

Sea buckthorn (Hippophae rhamnoides L.), as an economically and ecologically valuable plant with rich nutritional and bioactive compounds, has garnered significant interest. The demand for Sea buckthorn has explosive growth, highlighting the urgent need for the cultivation of fast-growing, high-quality Sea buckthorn seedlings. However, there are still some controversies in Sea buckthorn germplasm resource research. This review provides a comprehensive summary of the recent research findings on Sea buckthorn plants, encompassing their classification, distribution, propagation methods, medical functions, and valorization. It aims to offer strong support for the industrial utilization of the Sea buckthorn and explores the prospects for molecular breeding in Sea buckthorn.

Breeding early maturing cultivars is one of the most important objectives in pear breeding. Very early maturing pears provide an excellent parental material for crossing, but the immature embryo and low seed germination of their hybrid progenies often limit the selection and breeding of new early maturing pear cultivars. In this study, we choose a very early maturing pear cultivar ‘Pearl Pear’ as the study object and investigate the effects of cold stratification, the culture medium, and the seed coat on the germination and growth of early maturing pear seeds. Our results show that cold stratification (4 °C) treatment could significantly improve the germination rates of early maturing pear seeds. A total of 100 days of cold-temperature treatment in 4 °C and in vitro germination on White medium increased the germination rate to 84.54%. We also observed that seed coat removal improved the germination of early maturing pear seeds, with middle seed coat removal representing the optimal method, with a high germination rate and low contamination. The results of our study led to the establishment of an improved protocol for the germination of early maturing pear, which will greatly facilitate the breeding of new very early maturing pear cultivars.

BackgroundThe spatial distribution of tumor-infiltrating T cells and its dynamics during chemoradiotherapy combined with PD-1 blockade is little known in esophageal squamous cell carcinoma (ESCC).MethodsWe applied the multiplex immunofluorescence method to identify T cells (CD4+, CD8+ T cells, and their PD-1− or PD-1+ subsets) and myeloid-derived cells (CD11c+ dendritic cells, CD68+ macrophages, and their PD-L1+ subpopulations) in paired tumor biopsies (n = 36) collected at baseline and during combination (40 Gy of radiation) from a phase Ib trial (NCT03671265) of ESCC patients treated with first-line chemoradiotherapy plus anti-PD-1 antibody camrelizumab. We used the FoundationOne CDx assay to evaluate tumor mutational burden (TMB) in baseline tumor biopsies (n = 14). We dynamically assessed the nearest distance and proximity of T-cell subsets to tumor cells under combination and estimated the association between T-cell spatial distribution and combination outcome, myeloid-derived subsets, TMB, and patient baseline characteristics.FindingsWe found that the tumor compartment had lower T-cell subsets than the stromal compartment but maintained a comparable level under combination. Both before and under combination, PD-1− T cells were located closer than PD-1+ T cells to tumor cells; T cells, dendritic cells, and macrophages showed the highest accumulation in the 5–10-μm distance. Higher CD4+ T cells in the tumor compartment and a shorter nearest distance of T-cell subsets at baseline predicted poor OS. Higher baseline CD4+ T cells, dendritic cells, and macrophages were associated with worse OS in less than 10-μm distance to tumor cells, but related with better OS in the farther distance. Higher on-treatment PD-1-positive-expressed CD4+ and CD8+ T cells within the 100-μm distance to tumor cells predicted longer OS. T cells, dendritic cells, and macrophages showed a positive spatial correlation. Both high TMB and smoking history were associated with a closer location of T cells to tumor cells at baseline.ConclusionsWe firstly illustrated the T-cell spatial distribution in ESCC. Combining chemoradiotherapy with PD-1 blockade could improve the antitumor immune microenvironment, which benefits the treatment outcome. Further understanding the precision spatiality of tumor-infiltrating T cells would provide new evidence for the tumor immune microenvironment and for the combination treatment with immunotherapy.

In this study, we investigated the feasibility of Ginkgo biloba seeds powder by solid-state fermentation with Eurotium cristatum for developing high-value ginkgo seeds products. The optimum fermentation medium was consisted of 10 g of 40-mesh ginkgo seeds powder loaded in 100 mL Erlenmeyer flask with 50% (w/w) of water content, 4% (w/w) of MgSO4 and 5% (w/w) of KH2PO4 addition. The optimum fermentation conditions were pH 5.0, 2 × 108 CFU/g of inoculum size, 3 mL of sterilized water supplemented every two days during the four days of fermentation. Through fermentation, the spore number of E. cristatum was improved by about 36 times with the production of lovastatin reaching (54.10±0.16) µg/g. The antioxidant activity of fermented ginkgo seeds powder also got obvious enhancement, which could help eliminate excess free radicals produced by normal metabolism. The content of free amino acids increased by 82.32%. Except that the sugar was consumed in some degree, the other nutritional and functional components were well preserved while the content of detrimental ginkgolic acids was reduced by 44.97%. In addition, fermented ginkgo seeds powder possessed better digestibility and showed pleasant orange-like smelling. In conclusion, the quality of ginkgo seeds powder was remarkably improved through solid-state fermentation by using E. cristatum, which could be a promising way for functional applications of ginkgo seeds.

Ginkgo biloba L. is a tree species of significant economic and ecological importance. Prior studies of the Ginkgo biloba seed coat have predominantly focused on the sarcotesta and sclerotesta, with less attention paid to the endotesta. In this study, the development and formation of Ginkgo endotesta were examined using light microscopy and transmission electron microscopy. The structural properties of the mature endotesta were analyzed using micro-CT imaging and scanning electron microscopy. The results indicate that the endotesta possess a membranous structure primarily originating from the inner bead peridium, a segment of bead core tissue, and the macrospore membrane. The endotesta from the middle constriction line to the chalazal end comprises a single layer with a greyish-white papery structure. In contrast, the endotesta was divided into two inner and two outer layers, from the middle constriction line to the micropylar end. The outer endosperm adheres closely to the sclerotesta, while the inner endosperm adheres to the seed kernel. The surface of the endotesta was irregularly raised, with thicker wax at the chalazal end, whereas the micropylar end demonstrated similar characteristics with thinner wax and tumor layers. The endotesta contained 17 amino acids, 18 fatty acids, 10 trace elements, and 7 vitamins. Overall, its nutritional value was relatively well balanced.

Bilobalide exhibits numerous beneficial bioactivities, including neuroprotective, anti-inflammatory, and antioxidant activity. Our previous study demonstrated that bilobalide inhibits adipogenesis and promotes lipolysis. The dose-dependent cytotoxicity was found to be specific to the mature adipocytes only, indicating the potential for regulating apoptosis in them. Herein, we aimed to investigate the apoptotic effects of bilobalide on 3T3-L1 mature adipocytes and elucidate the underlying mechanisms thereof. Flow cytometry analysis (FACS) revealed the pro-apoptotic effects of bilobalide on these cells. Bilobalide induced early apoptosis by reducing the mitochondrial membrane potential (MMP). DNA fragmentation was confirmed using TUNEL staining. Additionally, bilobalide increased the intracellular reactive oxygen species (ROS) levels and activities of Caspases 3/9. Pre-treatment with NAC (an ROS scavenger) confirmed the role of ROS in inducing apoptosis. Moreover, bilobalide up- and down-regulated the expression of Bax and Bcl-2, respectively, at the mRNA and protein expression levels; upregulated the Bax/Bcl-2 ratio; triggered the release of cytochrome c from the mitochondria; and increased the protein expression of cleaved Caspase 3, cleaved Caspase 9, and PARP cleavage. These results support the conclusion that bilobalide induces apoptosis in mature 3T3-L1 adipocytes through the ROS-mediated mitochondrial pathway, and offers potential novel treatment for obesity.

Salicylic acid (SA), belonging to a family of naturally occurring phenolic compounds, is a crucial plant hormone involved in many biological processes, such as plant immunity, seed germination, root initiation, stomatal closure, and biotic/abiotic stress response. SA playing central roles in many metabolic processes have been extensively characterized. However, the function of SA in plant flower development has rarely been investigated. This paper reviews recent research advances on the roles of SA in flower development, including regulation of stamen development, flowering time, and ovarian development. This study provides multiple lines and levels of evidence substantiating that SA plays important roles in flower development, and suggests a pressing need to study the flower development-related functions of SA in both annual and perennial plants.

With the concept of sustainable management of plantations, individual trees with excellent characteristics in plantations have received attention from breeders. To improve and maintain long-term productivity, accurate and high-throughput access to phenotypic characteristics is essential when establishing breeding strategies. Meanwhile, genetic diversity is also an important issue that must be considered, especially for plantations without seed source information. This study was carried out in a ginkgo timber plantation. We used simple sequence repeat (SSR) markers for genetic background analysis and high-density terrestrial laser scanning for growth structural characteristic extraction, aiming to provide a possibility of applying remote sensing approaches for forest breeding. First, we analyzed the genetic diversity and population structure, and grouped individual trees according to the genetic distance. Then, the growth structural characteristics (height, diameter at breast height, crown width, crown area, crown volume, height to living crown, trunk volume, biomass of all components) were extracted. Finally, individual trees in each group were comprehensively evaluated and the best-performing ones were selected. Results illustrate that terrestrial laser scanning (TLS) point cloud data can provide nondestructive estimates of the growth structural characteristics at fine scale. From the ginkgo plantation containing high genetic diversity (average polymorphism information content index was 0.719) and high variation in growth structural characteristics (coefficient of variation ranged from 21.822% to 85.477%), 11 excellent individual trees with superior growth were determined. Our study guides the scientific management of plantations and also provides a potential for applying remote sensing technologies to accelerate forest breeding.

The diameter at breast height (DBH) of trees and stands is not only a widely used plant functional trait in ecology and biodiversity but also one of the most fundamental measurements in managing forests. However, systematically measuring the DBH of individual trees over large areas using conventional ground-based approaches is labour-intensive and costly. Here, we present an improved area-based approach to estimate plot-level tree DBH from airborne LiDAR data using the relationship between tree height and DBH, which is widely available for most forest types and many individual tree species. We first determined optimal functional forms for modelling height-DBH relationships using field-measured tree height and DBH. Then we estimated plot-level mean DBH by inverting the height-DBH relationships using the tree height predicted by LiDAR. Finally, we compared the predictive performance of our approach with a classical area-based method of DBH. The results showed that our approach significantly improved the prediction accuracy of tree DBH (R2 ​= ​0.85–0.90, rRMSE ​= ​9.57%–11.26%) compared to the classical area-based approach (R2 ​= ​0.80–0.83, rRMSE ​= ​11.98%–14.97%). Our study demonstrates the potential of using height-DBH relationships to improve the estimation of the plot-level DBH from airborne LiDAR data.

T cell receptor (TCR) repertoire as a biomarker for predicting immunotherapy efficiency has been widely studied. However, its dynamics during radiotherapy combined with PD-1 blockade is little known. Using paired tumor and blood samples from the phase Ib clinical study (NCT03222440), we investigate the time-spatial TCR repertoire in esophageal squamous cell carcinoma (ESCC) patients treated with first-line definitive radiotherapy concurrently with anti-PD-1 antibody camrelizumab, and also evaluate the association between TCR repertoire and clinical outcomes. TCR sequencing was performed on tumor biopsies (n = 34, 15 pairs) and peripheral CD8+ T cells (n = 36, 18 pairs) collected at baseline and during treatment (after 40 Gy radiation and 2 rounds of camrelizumab). Whole exome sequencing was applied to estimate genomic mutations and tumor mutation burden. We show that the intratumoral TCR repertoire at baseline was correlated with tumor microenvironment and presented heterogeneity inter-individually. T-cell clones inflowed mutually between tumors and peripheral blood under combination treatment, resulting in an elevation of intratumoral TCR diversity. The peripheral CD8+ TCR diversity at baseline, increased tumor-peripheral Morisita-Horn overlap during treatment, and expansion of persistent intratumoral T-cell clones during treatment predicted improved survival. While it is unclear whether radiation contributed to the TCR changes versus PD-1 therapy alone, our results firstly reveal radiotherapy combined with PD-1 blockade greatly promoted time-spatial alteration of TCR repertoire between tumor and peripheral blood, which demonstrate the peripheral CD8+ TCR diversity at baseline and dynamic alteration of intratumoral TCRs acted as potential effective biomarkers of radiotherapy combined with immunotherapy in ESCC.

Song, Fu, Yang, Niu, Tian, et al. present a high-quality genome assembly and transcriptome data of Taxus yunnanensis, a plant belonging to the Taxus genus known for sources of traditional medicine and natural anticancer drugs. This study explores the Taxus evolutionary history and sheds light on the Taxol biosynthetic pathway.

Background: The fruit Prunus mume has beneficial effects in the treatment of obesity and metabolic syndrome. However, its mechanism of action is unclear. Objective: We assessed the effect of a concentrated water extract of P. mume fruit (CEPM) on adipogenesis and beiging/browning in 3T3-L1 cells. Methods: The cell viability was determined by MTT assay. Lipid accumulation was assessed with Oil Red O (ORO) staining under different concentrations of CEPM. The effects of CEPM treatment during differentiation on beiging/browning and mitochondrial biogenesis in 3T3-L1 cells were investigated. Results: CEPM treatment suppressed differentiation and decreased lipid accumulation by downregulating the expression of key adipogenic genes, including PPARγ, C/EBPα, SREBP-1c, FAS, and perilipin A. In contrast, CEPM treatment increased the mitochondrial DNA (mtDNA) content and mRNA levels of mitochondrial biogenesis genes, including NAMPT, Nrf1, Nrf2, and CPT1α, and reduced reactive oxygen species levels. Importantly, CEPM increased the expression of brown/beige hallmark genes (Pgc-1α, Ucp1, Cidea, Cox7α1, Cox8b, Cd137, and Pdk-4), as well as proteins (UCP1, PGC-1α, NRF1, TBX1, and CPT1α). The high-performance liquid chromatography (HPLC) analysis reveals that CEPM contains mumefural, naringin, 5-HMF, citric acid, caffeic acid, and hesperidin. Conclusion: The first evidence we provided showed that CEPM has a dual role in 3T3-L1 cells inhibiting adipogenesis and promoting beiging/browning, and hence, could be a potential agent in the fight against obesity.

Ginkgo biloba, a relict plant, has been popularized and planted in most areas of China for its leaves, timber, and fruits. In the present study, the dynamic changes in leaf color, leaf pigment content during the color change period, and photosynthetic characteristics in different growth periods were studied to explore the coloring mechanism and adaptability of five late-deciduous superior Beijing G. biloba cultivars (LD1–LD5). The results showed that the leaf color change of each superior cultivar was relatively stable, and the discoloration period of LD3 and LD5 was later than that of others. From September to November, the chlorophyll a, chlorophyll b, and total chlorophyll content in all superior cultivars showed a downward trend, except in LD3, in which the pigment content was slightly higher in October than in September. Except in LD3 and LD4, the ratio of carotene content to total chlorophyll content in other cultivars slightly decreased in October. In May, the photosynthetic capacity of LD5 was stronger than that of other cultivars. The photosynthetic capacity of LD3 was strong in July and October. Our results imply that LD3 and LD5 are suitable for mixed planting with common G. biloba to increase the overall leaf color viewing period. Ginkgo biloba leaves turn yellow in autumn because of both a decrease in the chlorophyll content after leaf senescence and an increase in the Car content during leaf senescence. Although LD5 presented rapid seedling emergence, LD3 grew faster during the vigorous and late growth stages and is thus suitable for agricultural production.

Quercus gilva is an ecologically and economically important species of Quercus section Cyclobalanopsis and is a dominant species in evergreen broad-leaved forests in subtropical regions of East Asia. In the present study, we reported a high-quality chromosome-scale genome assembly of Q. gilva, the first reference genome for section Cyclobalanopsis, using the combination of Illumina and PacBio sequencing with Hi-C technologies. The assembled genome size of Q. gilva was 889.71 Mb, with a contig number of 773 and a contig N50 of 28.32 Mb. Hi-C scaffolding anchored 859.07 Mb contigs (96.54% of the assembled genome) onto 12 pseudochromosomes, with a scaffold N50 of 70.35 Mb. A combination of de novo, homology-based, and transcript-based predictions predicted a final set of 36,442 protein-coding genes distributed on 12 pseudochromosomes, and 97.73% of them were functionally annotated. A total of 535.64 Mb (60.20%) of repetitive sequences were identified. Genome evolution analysis revealed that Q. gilva was most closely related to Q. suber and they diverged at 40.35 Ma, and Q. gilva did not experience species-specific whole-genome duplication in addition to the ancient gamma (γ) whole-genome triplication event shared by core eudicot plants. Q. gilva underwent considerable gene family expansion and contraction, with 598 expanded and 6,509 contracted gene families detected. The first chromosome-scale genome of Q. gilva will promote its germplasm conservation and genetic improvement and provide essential resources for better studying the evolution of Quercus section Cyclobalanopsis.

Ginkgo biloba seed (GBS) contains rich nutrients, such as starch, protein, oil, and trace components, such as flavonoids and terpene lactones. Due to its high protein content, it can be used as a raw material for fermentation and brewing. In this study, GBS was selected as the object of a fermentation process optimization test. Six kinds of fermentation starter were selected to brew ginkgo wine. The results showed that different fermentation starters have significant impacts on the composition of the wine. The yeast group had higher total sugar content and comprehensive evaluation scores than the Jiuqu group, while the total acid and total free amino acid contents showed the opposite result. The total flavonoid and total terpene lactone contents of the yeast group were 21.0% and 12.8% higher than those of the Jiuqu group, respectively. However, the 4′-O-methylpyridoxine (MPN) and 4′-O-methylpyridoxine-5′-glucoside (MPNG) contents of the yeast group were also 12.6% and 2.3% higher than those of the Jiuqu group, respectively. The common volatile components in the two groups of samples were isoamyl alcohol, phenethyl alcohol, ethyl octanoate, and phenethyl acetate. The antioxidant capacity of ginkgo wine fermented by yeast was significantly higher than that of the Jiuqu group sample.

Due to today’s fast-paced lifestyle, most people are in a state of sub-health and face “unexplained fatigue”, which can seriously affect their health, work efficiency, and quality of life. Fatigue is also a common symptom of several serious diseases such as Parkinson’s, Alzheimer’s, cancer, etc. However, the contributing mechanisms are not clear, and there are currently no official recommendations for the treatment of fatigue. Some dietary polysaccharides are often used as health care supplements; these have been reported to have specific anti-fatigue effects, with minor side effects and rich pharmacological activities. Dietary polysaccharides can be activated during food processing or during gastrointestinal transit, exerting unique effects. This review aims to comprehensively summarize and evaluate the latest advances in the biological processes of exercise-induced fatigue, to understand dietary polysaccharides and their possible molecular mechanisms in alleviating exercise-induced fatigue, and to systematically elaborate the roles of gut microbiota and the gut-muscle axis in this process. From the perspective of the gut-muscle axis, investigating the relationship between polysaccharides and fatigue will enhance our understanding of fatigue and may lead to a significant breakthrough regarding the molecular mechanism of fatigue. This paper will provide new perspectives for further research into the use of polysaccharides in food science and food nutrition, which could help develop potential anti-fatigue agents and open up novel therapies for sub-health conditions.

The goal of this article is to conduct a detailed study of China’s forestry industry chain and determine the effect it has on the national economy. Based on the 2018 China Input–Output Table (153 departments) and other relevant data released by the National Bureau of Statistics, as well as the input–output method established by Wassily Leontief, this article conducts a comprehensive and detailed analysis of China’s forest industry chain and its impact on the national economy. The results show that the backward industrial chain of China’s forestry industry involves 105 industrial departments. China’s forestry sector purchases the most products from the tertiary industry, accounting for 56.05% of all forestry purchases. The products of the tertiary industry have a significant impact on forestry. The forward industry chain of China’s forestry involves 131 industrial sectors, with China’s forestry selling the most products to the secondary industry, accounting for 93.47% of all forestry sales. The forestry sensitivity coefficient is 0.767 and the influence coefficient is 0.65. The secondary industry has a significant impact on the sales of forestry products. The impact of the forward industry chain is greater than that of the backward industry chain and the impact of forestry on the national economy is quite large. Suggestions about how to expand the planting area and production scale of domestic economic forests were put forward. An increase in the added value of exported forestry products, improved service quality of the tertiary industry to forestry, and improved quality and quantity of products provided by forestry to the secondary industry are necessary.

As a “living fossil”, ginkgo (Ginkgo biloba L.) has significant ornamental, medicinal, and timber value. However, the breeding improvement of ginkgo was limited by the lack of enough excellent germplasms and suitable molecular markers. Here, we characterized numerous polymorphic insertion/deletion (InDel) markers using RAD-seq in 12 different ginkgo cultivars. The total of 279,534 InDels identified were unequally distributed across 12 chromosomes in the ginkgo genome. Of these, 52.56% (146,919) and 47.44% (132,615) were attributed to insertions and deletions, respectively. After random selection and validation, 26 pairs of polymorphic primers were used for molecular diversity analysis in 87 ginkgo cultivars and clones. The average values of observed heterozygosity and polymorphism information were 0.625 and 0.517, respectively. The results of population structure analyses were similar to those of neighbor-joining and principal component analyses, which divided all germplasms into two distinct groups. Moreover, 11 ginkgo core collections accounted for approximately 12.64% of the total ginkgo germplasms obtained, representing well the allelic diversity of all original germplasms. Therefore, these InDels can be used for germplasm management and genetic diversity analyses in ginkgo and the core collections will be used effectively for ginkgo genetic improvement.

Xanthoceras sorbifolium, belonging to the family Sapindaceae, has a beautiful tree shape, elegant leaves, large and many brightly colored flowers, and a long flowering duration. This plant is widely applied in gardens. In this study, 33 cultivars of Xanthoceras sorbifolium were selected from the perspective of ornamental properties, and their phenotypic traits, such as leaves, flowers, and branches, were measured and analyzed, and their phenotypic diversity was comprehensively evaluated using principal component analysis, in order to investigate the phenotypic diversity characteristics of Xanthoceras sorbifolium. The results showed that the genetic diversity index of the qualitative traits varied from 0.14 to 1.50, and that of quantitative traits varied from 1.76 to 2.05. The quantitative traits were more diverse than the qualitative traits. The coefficient of variation of the qualitative traits ranged from 16.90% to 57.96%, and that of quantitative traits ranged from 12.92% to 32.87%. The phenotypic traits of the tested cultivars had relatively rich variation. Furthermore, the level of the phenotypic diversity index of Xanthoceras sorbifolium was not consistent with the level of coefficient of variation, indicating large variation and uneven distribution of variation. Through principal component analysis, 17 quantitative characters were extracted into five principal components, with a cumulative contribution rate of 79.82%, representing the primary information on the quantitative characters of ornamental Xanthoceras sorbifolium cultivars. The F value of the 33 samples ranged from –2.79 to 1.93, and the comprehensive scores of seven cultivars were greater than 1, indicating that these cultivars had rich phenotypic diversity. Therefore, the screening, development, and utilization of fine germplasm resources of Xanthoceras sorbifolium should focus on these cultivars. The 33 cultivars were subsequently clustered into five categories through systematic clustering. The cluster analysis provided references for breeding ornamental Xanthoceras sorbifolium cultivars with different utilization values, such as large white flowers, small red flowers, large red flowers, large orange flowers, and double-petaled flowers.

Abstract Carpinus betulus and Carpinus putoensis are precious species in the world. Studies on the ecosystem function of the two species are rare. This study investigated the physiological and biochemical responses of C. betulus and C. putoensis to NO2 stress and their natural recovery. C. betulus and C. putoensis seedlings underwent fumigation with 12.0 mg/m3 NO2 for 0, 1, 6, 12, 24, 48, and 72 h, respectively. Then, the plants were allowed to recover at room temperature for 30 d. Physiological and biochemical changes in the leaves were compared between the two species. In terms of peroxidase (POD) activity, the damage response of C. betulus under NO2 stress appeared later than that of C. putoensis. The soluble protein content of C. betulus was noticeably higher than that of C. putoensis, and C. betulus exhibited more stable membrane lipoperoxidation. The tendency of the changes in nitrate reductase of C. betulus was less noticeable than that of C. putoensis. The variation amplitudes of N, K, Mg, Zn and Mn in the leaves of C. putoensis were greater than those of C. betulus. C. putoensis showed more sensitive metabolisms in response to NO2 stress compared with C. betulus. High-concentration NO2 caused damage to C. betulus and C. putoensis was reversible, and both species returned to normal growth via their own metabolism after 30-d recovery. The results of this study may provide useful reference data for quantitative assessment of the ecosystem function of C. betulus and C. putoensis and for their scientific application in urban greening.

Abstract Background Ginkgo (Ginkgo biloba L.) is an excellent landscape species. Its yellow-green leaf mutants are ideal materials for research on pigment synthesis, but the regulatory mechanism of leaf coloration in these ginkgo mutants remains unclear. Results We compared the metabolomes and transcriptomes of green and mutant yellow leaves of ginkgo over the same period in this study. The results showed that the chlorophyll content of normal green leaves was significantly higher than that of mutant yellow leaves of ginkgo. We obtained 931.52M clean reads from different color leaves of ginkgo. A total of 283 substances in the metabolic profiles were finally detected, including 50 significantly differentially expressed metabolites (DEMs). We identified these DEMs and 1361 differentially expressed genes (DEGs), with 37, 4, 3 and 13 DEGs involved in the photosynthesis, chlorophyll, carotenoid, and flavonoid biosynthesis pathways, respectively. Moreover, integrative analysis of the metabolomes and transcriptomes revealed that the flavonoid pathway contained the upregulated DEM (−)-epicatechin. Fourteen DEGs from the photosynthesis pathway were positively or negatively correlated with the DEMs. Conclusions Our findings suggest a complex metabolic network in mutant yellow leaves. This study will provide a basis for studies of leaf color variation and regulation.

Several dehydroabietylamine derivatives containing heterocyclic moieties such as thiophene and pyrazine ring were successfully synthesized. The antiproliferative activities of these thiophene-based Schiff-bases, thiophene amides, and pyrazine amides were investigated in vitro against Hela (cervix), MCF-7 (breast), A549 (lung), HepG2 (liver), and HUVEC (umbilical vein) cells by MTT assay. The toxicity of L1−L10 (IC50 = 5.92− >100 μM) was lower than L0 (1.27 μM) and DOX (4.40 μM) in every case. Compound L1 had higher anti-HepG2 (0.66 μM), anti-MCF-7 (5.33 μM), and anti-A549 (2.11 μM) and compound L3 had higher anti-HepG2 (1.63 μM) and anti-MCF-7 (2.65 μM) activities. Both of these compounds were recognized with high efficiency in apoptosis induction in HepG2 cells and intercalated binding modes with DNA. Moreover, with average IC50 values of 0.66 and 5.98 μM, L1 was nine times more effective at suppressing cultured HepG2 cells viability than normal cells (SI = 9). The relative tumor proliferation rate (T/C) was 38.6%, the tumor inhibition rate was up to 61.2%, which indicated that L1 had no significant toxicity but high anti-HepG2 activity in vivo. Thus, it may be a potential antiproliferation drug with nontoxic side effects.

Camptotheca acuminata Decne (Nyssaceae) (happy tree, tree of life, cancer tree) is a rapidly growing deciduous and endangered tree endemic to east Tibet and southern China (Liu et al., 2002). C. acuminata is an ornamental tree with monopodial growth, beautiful leaves and shade- and cold- tolerant in their natural environment. Recently, its demand has grown rapidly due to secondary metabolites production like Camptothecin and its derivatives with anti cancer and antivirus activity. The aim of the present study was to develop an efficient protocol for in vitro micropropagation of this valuable plant. Different cultural media based on both MS (Murashige and Skoog, 1962) or DKW (Driver and Kuniyuki, 1984) formulations with different cytokinins (6-benzylaminopurine, BAP or 2-isopentenyladenine, 2iP or meta-topolin, mT) have been involved. The aromatic cytokinin meta-topolin stimulated plant growth. The optimum multiplication rate of Camptotheca shoots occurred on the DKW basal medium, supplemented with 2.5 µM meta-topolin, grown under mixed LED light. The highest number of roots per plantlets was recorded on the rooting medium with 0.3 mgl-1 NAA.

BackgroundThe first clinical study (NCT03671265) of first-line chemoradiotherapy combined with PD-1 blockade showed promising treatment outcomes in locally advanced esophageal squamous cell carcinoma (ESCC). However, partial patients did not respond to the combination treatment. The roles of dendritic cells (DCs) and macrophages in this combination treatment remain poorly understood.MethodsWe performed multiplexed immunofluorescence method to identify CD11c+ DCs, CD68+ macrophages, and their PD-L1- or PD-L1+ subpopulations in paired tumor biopsies (n = 36) collected at baseline and during the combination treatment (after radiation, 40 Gy) from the phase Ib trial (NCT03671265). We applied whole exome sequencing in the baseline tumor biopsies (n = 14) to estimate tumor mutation burden (TMB). We dynamically investigated the spatial distribution of DCs and macrophages under chemoradiotherapy combined with PD-1 blockade, and evaluated the association between their spatial distribution and combination outcome, and TMB.ResultsThe results showed that high percentages of PD-L1- DCs and macrophages in the baseline tumor compartment, but not in the stromal compartment, predicted improved OS and PFS. Chemoradiotherapy combined with PD-1 blockade promoted DCs and macrophages to migrate closer to tumor cells. During combination treatment, PD-L1- tumor cells were nearest to PD-L1- DCs and macrophages, while PD-L1+ tumor cells were next to PD-L1+ DCs and macrophages. High TMB was closely associated with a shorter distance from tumor cells to DCs and macrophages. Shorter distance between PD-L1+ tumor cells and PD-L1+ DCs or PD-L1- macrophages during the combination was correlated with better OS. Shorter distance between PD-L1- tumor cells and PD-L1- macrophages during combination was associated with both longer OS and PFS.ConclusionsPD-L1- or PD-L1+ DCs and macrophages exhibit distinct spatial distribution in ESCC. The close distance between tumor cells and these antigen-presenting cells (APCs) is critical to the clinical outcome in chemoradiotherapy combined with PD-1 blockade in ESCC patients. Our results highlight the predictive potential of spatial patterns of APCs in chemoradiotherapy combined with immunotherapy and reveal the underlying mechanism of APCs participating in chemoradiotherapy-induced antitumor immune response in ESCC.

The Asian elephant (Elephas maximus Linnaeus) is a globally endangered species, an internationally protected species, and a first-class protected animal in China. However, future climate change and human activities exacerbate the instability of its habitat range, leading to a possible reduction in the range. By using multi-source remote sensing data and products, as well as climate change models, including ASTER GDEM v3, Landsat8 OLI image and ClimateAP, we examined the effects of ecological factors related to climate and natural and anthropogenic influences on the distribution of Asian elephants in Sipsongpanna. Multiyear elephant field tracking data were used with a MaxEnt species distribution model and the climate model. First, the distribution of Asian elephants in potentially suitable areas in Sipsongpanna was simulated under current climatic conditions without considering human activities. The predicted distribution was verified by existing Asian elephant migration trajectories. Subsequently, the distribution of potentially suitable areas for Asian elephants in Sipsongpanna was simulated under two climate change scenarios (RCP4.5, RCP8.5) in three periods (2025, 2055, and 2085). The changes in potentially suitable areas for Asian elephants in Sipsongpanna were analyzed under multiple climate change scenarios for the current (2017) and different future periods by considering the effects of human activities. The results show the following: (1) under anthropogenic interference (AI), the optimal MaxEnt model has a high prediction accuracy with the area under the curve (AUC) of 0.913. The feature combination (FC) includes linear, quadratic, and threshold features, and the regularization multiplier (RM) is 2.1. (2) Jackknife analyses of the non-anthropogenic interference (NAI) and anthropogenic interference (AI) scenarios indicate that topography (altitude (Alt)), temperature (mean warmest month temperature (MWMT)), and precipitation (mean annual precipitation (MAP)) are the top three factors influencing the distribution of Asian elephants. (3) The total area suitable for Asian elephants under current climate conditions and AI accounts for 46.35% of the total area. Areas of high suitability (occurrence probability >0.5) are located in Jinghong City in central Sipsongpanna and Mengla County in southeastern Sipsongpanna. Among them, the minimum habitat range and ecological corridors are mainly located in Mengman Town, Mohan Town, Mengla Town, Mengban Township, Dadugang Township, and Mengwang Township. (4) The change in potentially suitable areas for Asian elephants between current and future conditions is small under AI and large under undisturbed conditions.

There are some differences in the anti-inflammatory activities of four typical components in EGB (extracts of ginkgo biloba leaves), and there is also a synergistic relationship. The order of inhibiting the NO-release ability of single functional components is OA > GF > OPC > G. Ginkgolide (G), proanthocyanidins (OPC), and organic acids (OA) all have synergistic effects on ginkgo flavonoids (GF). GF:OA (1:9) is the lowest interaction index among all complexes, showing the strongest synergy. The anti-inflammatory mechanism of the compound affects the expression of p-JNK, p-P38, and p-ERK1/2 proteins by inhibiting the expression of iNOS and COX2 genes on NFKB and MAPK pathways. This also provides a research basis for the development of anti-inflammatory deep-processing products of EGB.

Abstract Background Phenotypic diversity of floral organs plays an important role in plant systematic taxonomy and genetic variation studies. Previous research have focused on the direction of variation but disregarded its degree. Phenotypic variation (including directions and degrees) of 17 floral traits from wild to cultivated crabapples were explored by comparing their distributions and deviations in three different dimensions: floral organ number, size, and the shape. Results Except for petal number, petal length / petal width, and sepal length / sepal width, the analyzed floral traits of cultivated crabapples all showed downward distributed box bodies in box plot analysis and left deviations of fitted curves in frequency distribution function analysis when compared to the wild, which revealed consistent variation directions of petaloid conversion (pistils or stamens → petals), size miniaturization (large → small), and shape narrowness (petal shape: circular → elliptic; sepal shape: triangular → lanceolate). However, only seven floral traits exhibited significant differences in box plot analysis, while all of the traits in frequency distribution function analysis were obviously offset. The variation degrees were quantitatively characterized by sizing traits > shaping traits > numbering traits and by horizontal dimensions > radial dimensions. Conclusions Frequency distribution function analysis was more sensitive than the box plot analysis, which constructed a theoretical basis for Malus flower type breeding and would provide a new quantitative method for future evaluation of floral variation among different groups of angiosperms at large.

To evaluate the anti-inflammatory function of mogroside V (MGV) and its molecular mechanism, dextran sulfate sodium (DSS)-induced colitis and lipopolysaccharide (LPS)-induced inflammation model were used in the study. Results indicated that MGV (100 mg/kg/day) supplementation in diet improved the health status of mice. Western blot results showed that MGV inhibited the expression pro-inflammatory factors of colonic tissues. MGV significantly reduced the expressions of pro-inflammatory cytokines in the LPS-stimulated RAW264.7 cells, also inhibited the phosphorylation of MAPKs. MGV promoted the activation of AMPK and inhibition the PI3K/Akt/mTOR pathway. Compound C (AMPK inhibitor) and SC79 (Akt activator) reversed the alleviating effect of MGV on the expression pro-inflammatory factors via a hub molecule of mTOR, indicating that MGV inhibits LPS-stimulated inflammation by triggering the AMPK-PI3K/Akt/mTOR pathway. Collectively, MGV can potentially be used in the treatments of ulcerative colitis, and it exerts anti-inflammatory effect via MAPK-NF-κB/AP-1 and AMPK-PI3K/Akt/mTOR pathways in ulcerative colitis.

Due to improved data collection and processing techniques, forestry surveys are now more efficient and accurate, generating large amounts of forestry data. Forestry Big Data (FBD) has become a critical component of the forestry inventory investigation system. In this study, publications on FBD were identified via the Web of Science database, and a comprehensive bibliometric analysis, network analysis, and analysis of major research streams were conducted to present an overview of the FBD field. The results show that FBD research only began nearly a decade ago but has undergone an upswing since 2016. The studies were mainly conducted by China and the US, and collaboration among authors is relatively fragmented. FBD research involved interdisciplinary integration. Among all the keywords, data acquisition (data mining and remote sensing) and data processing (machine learning and deep learning) received more attention, while FBD applications (forecasting, biodiversity, and climate change) have only recently received attention. Our research reveals that the FBD research is still in the infancy stage but has grown rapidly in recent years. Data acquisition and data processing are the main research fields, whereas FBD applications have gradually emerged and may become the next focus.

It has been reported that Celtis sinensis Pers. is employed as a folk medicine for the treatment of inflammatory diseases. But the mechanism supporting its use as anti-inflammatory remains unclear. To investigate the anti-inflammatory of Celtis sinensis Pers. ICR mice were provided Celtis sinensis Pers. leaf extract (CLE) at 100, 200 mg/kg after ginkgolic acids (GA) sensitization. Our data showed that CLE and the main flavonoid isovitexin in CLE could ameliorate GA-induced contact dermatitis in mice. Ear swelling, inflammatory cell infiltration and splenomegaly were inhibited significantly by isovitexin, while the weight loss of mice in the isovitexin-treated group was much better than that in the dexamethasone-treated group (positive control drug). It has been reported in previous research that GA-induced inflammation is closely related to the T cell response. Therefore, T cells were the focus of the anti-inflammatory effect of isovitexin in this paper. The in vivo results showed that isovitexin (10, 20 mg/kg) inhibited the expression of proinflammatory cytokines (TNF-α, IFN-γ, IL-2 and IL-17A) in lymph nodes, inhibited the secretion of cytokines into the serum from mice with contact dermatitis and promoted the expression of apoptosis-related proteins. In vitro, isovitexin also induced apoptosis and inhibited proinflammatory cytokine expression in Con A-activated T cells. Further study showed that the MAPK and STAT signaling pathways and the phosphorylation of SHP2 were inhibited by isovitexin. Both molecular docking and biological experiments indicated that SHP2 may be an anti-inflammatory target of isovitexin in T cells. Taken together, isovitexin can serve as a potential natural agent for the treatment or prevention of GA-induced inflammatory problems.

Abstract Background Long noncoding RNAs (lncRNAs) play an important role in diverse biological processes and have been widely studied in recent years. However, the roles of lncRNAs in leaf pigment formation in ginkgo (Ginkgo biloba L.) remain poorly understood. Results In this study, lncRNA libraries for mutant yellow-leaf and normal green-leaf ginkgo trees were constructed via high-throughput sequencing. A total of 2044 lncRNAs were obtained with an average length of 702 nt and typically harbored 2 exons. We identified 238 differentially expressed lncRNAs (DELs), 32 DELs and 49 differentially expressed mRNAs (DEGs) that constituted coexpression networks. We also found that 48 cis-acting DELs regulated 72 target genes, and 31 trans-acting DELs regulated 31 different target genes, which provides a new perspective for the regulation of the leaf-color mutation. Due to the crucial regulatory roles of lncRNAs in a wide range of biological processes, we conducted in-depth studies on the DELs and their targets and found that the chloroplast thylakoid membrane subcategory and the photosynthesis pathways (ko00195) were most enriched, suggesting their potential roles in leaf coloration mechanisms. In addition, our correlation analysis indicates that eight DELs and 68 transcription factors (TFs) might be involved in interaction networks. Conclusions This study has enriched the knowledge concerning lncRNAs and provides new insights into the function of lncRNAs in leaf-color mutations, which will benefit future selective breeding of ginkgo.