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Gleditsia sinensis thorns (GST) have been used as a traditional medicine for carbuncles and skin diseases. The purpose of this study was to decide whether non-toxicological levels of water extract of GST (WEGST) are effective in inhibiting the progress of prostate cancer formation and to identify the target molecule involved in the WEGST-mediated inhibitory process of prostate cancer cell migration and in vivo tumor formation. Through the Boyden chamber migration assay, we found that non-toxic levels of WEGST could not attenuate the PC3 migration to the bottom area coated with serum but significantly inhibited PC3 cell migration to the collagen-coated bottom area. We also found that non-toxic levels of WEGST significantly attenuated collagen against adhesion. Interestingly, ectopic administration of WEGST could not affect the expression of alpha 2 beta 1 integrin, which is known as a receptor of collagen. However, when the PC3 cells adhered to a collagen-coated plate, the expression of alpha 2 integrin but not that of beta 1 integrin was significantly inhibited by the administration of non-toxic levels of WEGST, leading to the inhibition of focal adhesion kinase (FAK) phosphorylation. Furthermore, oral administration of WEGST (25 mg/kg/day) significantly inhibited the size of a PC3 cell-xenografted tumor. Taken together, these results suggest a novel molecular mechanism for WEGST to inhibit prostate cancer progression at particular stages, such as collagen-mediated adhesion and migration, and it might provide further development for the therapeutic use of WEGST in the treatment of prostate cancer progression.

Background Gleditsia sinensis Lam. (Leguminosae), a dioecious perennial arbor, demonstrates important medicinal properties and economic value. These properties can be harnessed depending on the sex of the plant. However, the sex of the plants is difficult to identify accurately through morphological methods before the flowering. Results We used bulked segregant analysis to screen sex-specific simple sequence repeat (SSR) markers in G. sinensis . Five male and five female plants were pooled to form the male and female bulks, respectively, and subjected to whole-genome sequencing. After high-throughput sequencing, 5,350,359 sequences were obtained, in which 2,065,210 SSRs were searched. Among them, the number of duplicated SSRs was the highest. The male plants could reach 857,874, which accounted for 60.86% of the total number of male plants. The female plants could reach 1,447,603, which accounted for 56.25% of the total model of the female plants. Among all the nucleotide repeat types, the A/T-rich motif was the most abundant. A total of 309,516 female strain-specific SSRs were selected by clustering. After designing the primers, the male and female gene pools were amplified, and five pairs of primers (i.e., 27, 34, 36, 39, and 41) were found to amplify the differential bands in the male and female gene pools. Using the five pairs of primers, we performed PCR verification on 10 individuals of known sex, which constructed the gene pool. The female plants amplified a single fragment of lengths (i.e., 186, 305, 266, 203, and 260 bp) and no male plant strip, thereby completing the identification of the male and female sexes of the G. sinensis. Conclusions This study provides accurate sex identification strategies between female and male plants, thus improving the utilization rate of G. sinensis resources.

This study aims to explore the low phosphorus (P) tolerance of saplings from different Gleditsia sinensis Lam. families. It also seeks to screen for Gleditsia sinensis families with strong low P tolerance and identify key indicators for evaluating their tolerance. This research provides a foundation for the breeding of superior families of Gleditsia sinensis and the study of mechanisms underlying low P tolerance. Using saplings from 30 Gleditsia sinensis families as the research subjects, a sand culture pot experiment was conducted. This study set up low P treatment (0.01 mmol L −1 ) and normal P treatment (1 mmol L −1 ). Twenty-five indicators including growth morphology, biomass, root morphology, and P content were measured. The low P tolerance coefficient was used as the basic data for assessing the low P tolerance of Gleditsia sinensis . The fuzzy comprehensive evaluation method was employed to comprehensively assess the low P tolerance types of Gleditsia sinensis a stepwise regression model was established to identify the key evaluation indicators for low P tolerance. The results indicate that low P stress reduced plant height, stem diameter, and biomass in most Gleditsia sinensis families, but increased the root morphological indicators, root-shoot ratio and PUE of various organs. Principal component analysis transformed the 25 indicators into 6 independent comprehensive indicators, with a cumulative contribution rate of 86.743%. The fuzzy comprehensive evaluation method calculated a comprehensive evaluation value (D value), enabling the screening of Gleditsia sinensis families into low P tolerant and low P sensitive types. Cluster analysis grouped the 30 Gleditsia sinensis families into 4 types. Among them, F13, F10, F9, F18, F15, and F28 were classified as low P tolerant types; F6, F23, F3, F17, F20, F2, F12, F11, F16, F8, F5, F27, F1, and F26 were categorized as intermediate types; F30, F7, F22, F4, F19, F29, F24, F14 and F25 were considered low P sensitive types, and F21 was classified as extremely low P sensitive types. The stepwise regression analysis identified the indicators stem diameter, total root volume, shoot dry weight, total root projection area, and leaf P content as the key factors for discriminating the low P tolerance of Gleditsia sinensis . The regression model is as follows: D=-0.005 + 0.323 stem diameter *+0.154 * total root volume + 0.196* shoot dry weight + 0.139* total root projection area − 0.112* leaf P content. In summary, low P stress inhibited the growth of Gleditsia sinensis saplings, but it increased the root morphological indicators, root-shoot ratio and PUE of various organs to cope with low P environments. The screening identified F13, F10, F9, F18, F15, and F28 as low P tolerant Gleditsia sinensis families. The evaluation indicators for low P tolerance in Gleditsia sinensis were identified as stem diameter, total root volume, shoot dry weight, total root projection area and leaf P content.

Gleditsia sinensis is a valuable tree species with important pharmaceutical uses. However, high soil NaCl concentration limits its growth in saline soil, including coastal areas. This study aimed to investigate the effects of arbuscular mycorrhizal fungi (AMF) on G. sinensis salinity tolerance and reveal its underlying physiological mechanism. A greenhouse experiment was performed. G. sinensis seedlings with and without AMF inoculation were subjected to four salinity levels: 0- (control), 50-, 100-, and 150-mM NaCl. After 2 months, the seedlings were harvested and analyzed for growth and biochemical parameters (antioxidant enzyme activity, superoxide anion content, malondialdehyde content, nutrient concentration, ion concentration, etc.). High AMF colonization rates (over 95%) and high mycorrhizal dependencies (over 75%) were observed across all NaCl levels. AMF effectively enhanced the salinity tolerance of G. sinensis seedlings by enhancing leaf stomatal conductance inducing higher net photosynthetic rates; improving peroxidase, catalase, and superoxide dismutase activities resulting in higher membrane stability indexes, lower superoxide anion and malondialdehyde contents in leaves and roots; increasing P/N ratio to mitigate P-limited biomass products; selectively absorbing less Na+ and more Ca2+ in their tissues to alleviate ion toxicity and maintain more favorable ion balances (e.g., K+/Na+) in their tissues. The results suggested AMF could effectively improve the salinity tolerance of G. sinensis, owning the great potential for afforestation and rehabilitation of G. sinensis in coastal areas.

As an economically important tree, Gleditsia sinensis Lam. is widely planted. A lack of background genetic information on G. sinensis hinders molecular breeding. Based on PacBio single-molecule real-time (SMRT) sequencing and analysis of G. sinensis , a total of 95,183 non-redundant transcript sequences were obtained, of which 93,668 contained complete open reading frames (ORFs), 2,858 were long non-coding RNAs (LncRNAs) and 18,855 alternative splicing (AS) events were identified. Genes orthologous to different Gleditsia species pairs were identified, stress-related genes had been positively selected during the evolution. AGA, AGG, and CCA were identified as the universal optimal codon in the genus of Gleditsia . EIF5A was selected as a suitable fluorescent quantitative reference gene. 315 Cytochrome P450 monooxygenases ( CYP450s ) and 147 uridine diphosphate (UDP)-glycosyltransferases ( UGTs ) were recognized through the PacBio SMRT transcriptome. Randomized selection of GsIAA14 for cloning verified the reliability of the PacBio SMRT transcriptome assembly sequence. In conclusion, the research data lay the foundation for further analysis of the evolutionary mechanism and molecular breeding of Gleditsia .

Gleditsia sinensis is a dioecious perennial tree, with its fruit size directly impacting its economic value. Investigating the feasibility of hybridization between different Gleditsia varieties and the influence of parentage on fruit size is crucial. In this study, we compared the pollen viability of Gleditsia sinensis , Gleditsia japonica var. delavayi ( G. delavayi ), Gleditsia japonica and Gleditsia fera under long-term storage, and explored the feasibility of intraspecific and interspecific hybridization of Gleditsia sinensis . We also evaluated the genetic characteristics of parentage on the fruit size and fruit set rate of Gleditsia sinensis . The results indicated that dry storage at -80 °C is suitable for the long-term preservation of pollen for most Gleditsia varieties. Interspecific hybridization of G. sinensis is feasible, and hybridization wit h G. fera exhibits a high general combining ability for fruit size. Therefore, G. fera can serve as an excellent male parent for interspecific hybridization with G. sinensis . The phenotypic traits of the current generation’s pods are primarily influenced by the additive effects of the maternal parent and the non-additive effects of both parents. Pod length and width exhibit high heritability, while pod thickness has lower heritability and is more significantly influenced by non-additive effects.

The Lam. polysaccharide gum contained in seeds is not only an additive specified in national food standards, but also an important strategic resource for industrial raw materials such as oil and natural gas in China.The main component of polysaccharide gum is galactomannan (GM). To date, most studies have focused on the structural modification and component separation of the polysaccharide gum of seeds, with only a few reports on the regulatory genes involved in its formation. Transcriptome analysis was performed to assess the gene networks associated with GM synthesis in seeds at four stages: 6, 9, 12, and 16 weeks after flowering. The result indicated that differential expression analysis identified 20 unigenes linked to five critical enzymes in the GM biosynthesis pathway. Further pathway enrichment analysis demonstrated that fructokinase, galactose gyrase, inositol galactoside synthase, phosphogalactosyltransferase, and raffinose synthase play pivotal roles in GM biosynthesis, positively regulating its production. The results of this study provide new ideas for the research of GM biosynthesis related genes in and enhance the potential application prospects in genetic engineering.

The seeds of Gleditsia sinensis and Gleditsia microphylla, widespread in China, are an important source of galactomannans. G. sinensis gum (GSG) and G. microphylla gum (GMG) were purified and precipitated using different concentrations of ethanol and isopropanol. The GSG and GMG, precipitated in different stages, presented different characteristics, including polymer recovery, mannose/galactose ratio, chemical composition, molecular weight, and morphological appearance. The galactomannan recovery of GSG and GMG in 33.3% ethanol was 81.7% and 82.5%, respectively, while that in 28.8% isopropanol was 81.3% and 82.9%, respectively. To achieve similar precipitation efficiency, the amount of isopropanol should be lower than that of ethanol because of the lower dielectric constant of isopropanol (20 vs. 25 for ethanol). The precipitation behavior of galactomannans in polar organic solvents was dependent on the molecular structures and properties of the solvent. A higher mannose/galactose ratio and a higher molecular weight was obtained in a lower concentration of alcohols.

Background Gleditsia sinensis Lam. (Fabaceae) is a medicinal legume characterized by its spines and pods, which are rich in saponins, polysaccharides, and various specialized metabolites with potential medicinal and industrial applications. The low fruit set rate in artificially cultivated economic forests significantly impedes its development and utilization. A comprehensive understanding of the cellular events, physiological and biochemical processes, and molecular regulatory mechanisms underlying fruit initiation and early fruit development is essential for enhancing yield. However, such information for G. sinensis remains largely unexplored. Results In this study, we identified that the early fruit development process in G. sinensis can be categorized into three distinct stages: pollination, the critical period of fertilization, and the initial fruit development followed by subsequent growth. The dynamic changes in non-structural carbohydrates and endogenous plant hormones within the ovary were found to play a significant role during fruit set and the early stages of fruit development. Additionally, the high activity of gibberellin, cytokinin, and sucrose-metabolizing enzymes in the ovary was conducive to early fruit development. Furthermore, we generated high-resolution spatiotemporal gene expression profiles in the ovary from the stage of efflorescence to early fruit development. Comparative transcriptomics and weighted gene co-expression network analysis revealed specific genes and gene modules predominant at distinct developmental stages, thereby highlighting unique genetic programming. Overall, we identified the potential regulatory network governing fruit initiation and subsequent development, as well as the sets of candidate genes involved, based on the aforementioned results. Conclusions The results offer a valuable reference and resource for the application of exogenous substances, such as hormones and sugars, during critical fruit development periods, and for the development of molecular tools aimed at improving yield.

G. sinensis thorn (called “zào jiǎo cì”, ZJC) has important medicinal and economic value, however, little is known about the molecular mechanisms behind the development of ZJC. In this study, we measured the content of soluble sugar and starch during the growth and development of the thorn, and performed transcriptome sequencing of the thorn segment, non-thorn segment, apex, and root tip at five distinct stages of thorn formation. The results showed that, with the growth of ZJC, the soluble sugar content of the roots, hypocotyls, thorn stems, thornless stems, leaves, and the starch content of the roots and leaves all firstly increased and then decreased after the basic structure of thorns was formed; the soluble sugar content and starch content of ZJC showed an overall downward trend (decreased by 59.26% and 84.56%, respectively). Myb-like, YABBY2, Growth-regulating factor 3, TCP2, Zinc transporter 8, and another 25 genes may be related to the maintenance and growth of thorns. Gene Ontology (GO) enrichment analysis of differentially expressed genes (DEGs) between stems with thorn and thorn-free stems found that a significant number of DEGs were annotated with terms related to the positive regulation of development, heterochronic (GO:0045962), the positive regulation of photomorphogenesis (GO:2000306), and other biological process (BP) terms. The developmental initiation regulation of ZJC may be regulated by TCP transcription factors (TFs). Eight genes were selected randomly to validate the RNA-seq results using real-time quantitative PCR (RT-qPCR) and they indicated that the transcriptome data were reliable. Our work provided a comprehensive review of the thorn development of G. sinensis.