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Background Polyploidy is a key driver of germplasm innovation in plants, contributing significantly to speciation and the evolution of cultivated crops. Trichosanthes cucumerina Linn. is an economically important crop, valued for its edible, medicinal, and industrial properties. This study aims to explore new germplasm resources of T. cucumerina with desirable traits such as high yield and stress resistance by analyzing and comparing the morphological characteristics, anatomical traits, and root powder microscopic features of diploid and autotetraploid T. cucumerina . Results (1) The most effective method for inducing autotetraploidy was soaking explants in a 0.05% colchicine solution for 72 h, which resulted in the highest explant survival rate (70.67%) and autotetraploid induction rate (31.33%), yielding 145 autotetraploid plants. (2) Flow cytometry and root-tip chromosome counting confirmed that diploid plants had a chromosome number of 2n = 2x = 22, while autotetraploid plants had 2n = 4x = 44. (3) On MS medium supplemented with 6-BA (3.0 mg/L), KT (3.0 mg/L), NAA (1.0 mg/L) and 360.36 mg/L Ca²⁺, autotetraploid stem segments exhibited the highest shoot induction, with a proliferation coefficient of 6.43. (4) The optimal rooting medium for autotetraploid plantlets consisted of MS medium supplemented with 1.0 mg/L NAA, achieving a 100% rooting rate. Following acclimatization and transplantation of regenerated autotetraploid plantlets, the survival rate reached 92%. (5) Compared to diploid plants, autotetraploid plants displayed significantly larger nutritional organs (leaves, stems, roots), enhanced growth parameters (leaf width, stomatal length, stomatal width, root fresh weight, and root dry weight), and more developed cellular structures (palisade tissue, spongy tissue, vascular tissue, starch granules, parenchyma cells, and cork cells). Conclusion This study successfully induced autotetraploidy in T. cucumerina and established an efficient artificial regeneration system. Compared to diploid plants, autotetraploid plants demonstrated significant improvements in morphological characteristics, anatomical traits, and growth parameters. These findings provide valuable resources for breeding new varieties and advancing genetic research on T. cucumerina .

Key messageDe novo transcriptome analysis from callus, leaf, and fruit of Trichosanthes cucumerina L. for the identification of genes associated with triterpenoid biosynthesis, especially bryonolic acid and cucurbitacin B.Trichosanthes cucumerina L. (TC) has been used as a medicinal plant in Thailand with various potential functions. Two major triterpenoids found in this plant, bryonolic acid and cucurbitacin B, are receiving increased attention for their activities. Here, we provide TC transcriptome data to identify genes involved in the triterpenoid biosynthetic pathway through callus, where was previously suggested as a novel source for bryonolic acid production as opposed to leaf and fruit. A de novo assembly of approximately 290-thousand transcripts generated from these tissues led to two putative oxidosqualene cyclases: isomultiflorenol synthase (IMS) and cucurbitadienol synthase (CBS). TcIMS and TcCBS, genes that encode substrates for two characteristic triterpenoids in cucurbitaceous plants, were identified as isomultiflorenol synthase and cucurbitadienol synthase, respectively. These two genes were functionally characterised in mutant yeast Gil77 systems, which led to the productions of isomultiflorenol and cucurbitadienol. Moreover, the callus-specific gene expression profiles were also presented. These obtained information showed candidate cytochrome P450s with predicted full-length sequences, which were most likely associated with triterpenoid biosynthesis, especially bryonolic acid. Our study provides useful information and a valuable reference for the further studies on cucurbitaceous triterpenoids.

Background Cucurbitaceae is an economically important family with a wide geographical distribution range. The phylogenetic backbone of Cucurbitaceae has not been well resolved, and conflicting results have been reported in previous phylogenetic studies. Trichosanthes is the largest genus in Cucurbitaceae; however, its infrageneric classification remains controversial. Here, we used both plastome and nuclear datasets to reconstruct the phylogeny of Cucurbitaceae with emphasis on Trichosanthes . The plastome dataset includes 137 taxa, representing 15 tribes and 35 genera. The 1,244 single-copy nuclear genes (SCNs) dataset includes 143 samples from all 15 tribes and 68 genera of Cucurbitaceae. Results Phylograms of plastome dataset and 1,244 SCNs essentially agreed with each other with the exception of first branching tribes and Tr. Bryonieae. The phylogram of the plastome dataset was used to illustrate phylogeny and molecular dating. Cucurbitaceae was subdivided into nine clades along the phylogram backbone. The first branching clade of Cucurbitaceae included four tribes: Actionstemmateae, Fevilleeae, Gomphogyneae, and Zanonieae. Actionstemmateae was sister to the remaining three tribes, with Gomphogyneae and Fevilleeae being successive sister to Zanonieae. Six tribes, including Indofevilleeae, Thladiantheae, Momordiceae, Siraitieae, Joliffieae, and Bryonieae, formed successive and well-supported clades along the backbone of Cucurbitaceae. Cucurbitaceae originated in the Late Cretaceous (ca. 94.186 million years ago [MYA]), and diversified during the Late Cretaceous to the Eocene. Diversification of all tribes occurred during the later Cretaceous to Eocene. Trichosanthes originated ca. 30.089 MYA, and the section diversification occurred in middle Miocene approximately 6 million years (20.839 − 14.804 MYA). Climate optimum in the Mid-Eocene and Mid-Miocene was probably among the important drivers of the diversification of Trichosanthes complex. Conclusions Our results provide new insights into the phylogeny of Cucurbitaceae, including members of the first branching clade and the relative phylogenetic positions of Indofevilleeae, Thladiantheae, and Momordiceae, and show that Trichosanthes is paraphyletic with three genera nested within it. Our results suggest that the previously proposed infrageneric classification systems for Trichosanthes need to be revised. Section Involucraria was subdivided into two distantly related groups. Trichosanthes originated in the Oligocene and experienced rapid diversification in the Middle Miocene and throughout the Pliocene. Long-fringed petals might have arisen once in a clade formed by Trichosanthes and Hodgsonia.

Three grinding stones from Shizitan Locality 14 (ca. 23,000-19,500 calendar years before present) in the middle Yellow River region were subjected to usewear and residue analyses to investigate human adaptation during the last glacial maximum (LGM) period, when resources were generally scarce and plant foods may have become increasingly important in the human diet. The results show that these tools were used to process various plants, including Triticeae and Paniceae grasses, Vigna beans, Dioscorea opposita yam, and Trichosanthes kirilowii snakegourd roots. Tubers were important food resources for Paleolithic hunter-gatherers, and Paniceae grasses were exploited about 12,000 y before their domestication. The long tradition of intensive exploitation of certain types of flora helped Paleolithic people understand the properties of these plants, including their medicinal uses, and eventually led to the plants' domestication. This study sheds light on the deep history of the broad spectrum subsistence strategy characteristic of late Pleistocene north China before the origins of agriculture in this region.

To analyze the effects of different cultivation patterns on the structure and diversity of the microbial community in the rhizosphere soil of ( ) arms to establish reasonable and effective strategies to mitigate the continuous cropping barriers and promote the high-quality cultivation of . Three distinct cultivation patterns were investigated: open-field cultivation (TM1), film-mulched cultivation (TM2), and soybean intercropping cultivation (TM3). High-throughput sequencing and bioinformatic analyses were employed to evaluate the rhizosphere microbiome, and redundancy analysis was utilized to investigate the relationship between the microbial communities and soil nutrient indicators. TM2 and TM3 increased soil bacterial community diversity, reduced fungal community diversity, elevated the relative abundance of beneficial bacterial genera, and reduced the abundance of detrimental fungal genera in the rhizosphere soil. The relative abundance of , , and in TM2 and TM3 was markedly higher than in TM1. Conversely, the relative abundance of , , , and in TM2 and TM3 was significantly reduced compared to TM1. The contents of available potassium (AK), total nitrogen (TN), total phosphorus (TP), and pH in the rhizosphere soil of TM2 and TM3 were significantly higher than those in TM1. The distribution of soil bacterial genera was significantly influenced by the contents of TN and AK, while the distribution of soil fungal genera was significantly or extremely significantly impacted by the contents of TP, total potassium (TK), soil organic matter (SOM), and pH. The content of AK was extremely significantly positively correlated with the relative abundance of , whereas the content of TK showed an extremely negative correlation with the relative abundance of . Similarly, pH demonstrated an extremely negative correlation with the relative abundance of and . Film-mulched cultivation and soybean intercropping cultivation altered the soil nutrients, as well as the structure and diversity of soil microbial communities. Thus, in agricultural production, film-mulched cultivation and soybean intercropping cultivation can serve to regulate soil nutrients and microbial communities, thereby mitigating the barriers of continuous cropping of .

The present research focuses on developing an innovative biochar-based heterogeneous catalyst from Prosopis Juliflora biomass waste using response surface methodology and genetic algorithm (GA) to optimize pyrolysis parameters, achieving a 46.31% PJBC yield from 60 mg of biomass at 790 °C for 60 min. The pyrolyzed PJBC is characterized using SEM, FTIR, XRD, EDX, BET, XPS analyses, and physico-chemical measurements to confirm its catalytic activity. Now, the newly synthesized PJBC serves as an efficient catalyst for waste Trichosanthes cucumerina seed biodiesel (WTSB) production from waste Trichosanthes cucumerina seed bio-oil through trans-esterification, achieving a maximum yield of 97.42%. Also, the WTSB exhibits excellent physico-chemical properties that meet most of the ASTM D6751 standards for biodiesel and closely align with the characteristics of conventional diesel fuel. Therefore, this research utilized neat WTSB and WTSB/diesel blends (WTSB25, WTSB50, and WTSB75) in a direct injection (DI) diesel engine at variable load settings. Among all WTSB blends, the WTSB25 blend showed closer variations of 1.65% lower BTE and 9.29% higher BSEC when compared to conventional diesel fuel readings. Its peak in-cylinder pressure and heat release rate were similar to those of diesel fuel at 100% engine load. Emission analysis indicated that the WTSB25 reduced specific HC, CO, and smoke opacity emissions by 8.39%, 13.97%, and 4.18%, respectively. However, specific NO emissions increased slightly by 3.05% compared to diesel fuel. Thus, WTSB25 is validated as a viable diesel alternative requiring no significant engine modifications. The environmental impact, lifecycle and economic feasibility are also discussed.

Trichosanthes kirilowii ( T . kirilowii ) is a valuable plant used for both medicinal and edible purposes. It belongs to the Cucurbitaceae family. However, its phylogenetic position and relatives have been difficult to accurately determine due to the lack of mitochondrial genomic information. This limitation has been an obstacle to the potential applications of T . kirilowii in various fields. To address this issue, Illumina and Nanopore HiFi sequencing were used to assemble the mitogenome of T . kirilowii into two circular molecules with sizes of 245,700 bp and 107,049 bp, forming a unique multi-branched structure. The mitogenome contains 61 genes, including 38 protein-coding genes (PCGs), 20 tRNAs, and three rRNAs. Within the 38 PCGs of the T . kirilowii mitochondrial genome, 518 potential RNA editing sites were identified. The study also revealed the presence of 15 homologous fragments that span both the chloroplast and mitochondrial genomes. The phylogenetic analysis strongly supports that T . kirilowii belongs to the Cucurbitaceae family and is closely related to Luffa. Collinearity analysis of five Cucurbitaceae mitogenomes shows a high degree of structural variability. Interestingly, four genes, namely atp 1, ccm FC, ccm FN, and mat R, played significant roles in the evolution of T . kirilowii through selection pressure analysis. The comparative analysis of the T . kirilowii mitogenome not only sheds light on its functional and structural features but also provides essential information for genetic studies of the genus of Cucurbitaceae .

Fruit length is a crucial agronomic trait of snake gourd ( Trichosanthes anguina L); however, genes associated with fruit length have not been characterised. In this study, F 2 snake gourd populations were generated by crossing the inbred lines, S 1 and S 2 (fruit lengths: 110 and 20 cm, respectively). Subsequently, bulk segregant analysis, sequencing, and fine-mapping were performed on the F 2 population to identify target genes. Our findings suggest that the fruit length of snake gourd is regulated by a major-effect regulatory gene. Mining of genes regulating fruit length in snake gourd to provide a basis for subsequent selection and breeding of new varieties. Genotype-phenotype association analysis was performed on the segregating F 2 population comprising 6,000 plants; the results indicate that the target gene is located on Chr4 (61,846,126–61,865,087 bp, 18.9-kb interval), which only carries the annotated candidate gene, Tan0010544 (designated TFL ). TFL belongs to the MADS-box family, one of the largest transcription factor families. Sequence analysis revealed a non-synonymous mutation of base C to G at position 202 in the coding sequence of TFL , resulting in the substitution of amino acid Gln to Glu at position 68 in the protein sequence. Subsequently, an InDel marker was developed to aid the marker-assisted selection of TFL. The TFL in the expression parents within the same period was analysed using quantitative real-time PCR; the TFL expression was significantly higher in short fruits than long fruits. Therefore, TFL can be a candidate gene for determining the fruit length in snake gourd. Collectively, these findings improve our understanding of the genetic components associated with fruit length in snake gourds, which could aid the development of enhanced breeding strategies for plant species.

Polysaccharides from Trichosanthes peel (TPP) were obtained by ultrasound-assisted extraction. TPP-1 was separated from the TPP by Sephadex G-100 column chromatography. Phosphorylation of TPP-1 was carried out and phosphorylated TPP-1 was named as PTTP-1. The results of infrared spectra, 13C nuclear magnetic resonance spectra and 31P nuclear magnetic resonance spectra showed that the main structure of PTPP-1 was similar to that of TPP-1 and -H2PO3 groups which were conjugated to C-6 of →4)-α-D-Manp-(1→, C-4 of →6)-α-D-Galp-(1→, C-2 and C-3 of →1)-α-L-Araf, C-2 of →1)-α-L-Araf-(3→, and C-6 and C-3 of →1)-α-D-Glcp. In vivo antiaging activity results proved that TTP-1 and PTTP-1 could both significantly improve the body weight, spleen index, and thymus index of the D-galactose-induced aging mice, increase the levels of superoxide dismutase, catalase, glutathione peroxidase, and reduce malondialdehyde contents in the liver, brain, and serum of aging mice. These results indicated that both TPP-1 and PTTP-1 presented significant antiaging activity. Moreover, PTTP-1 showed stronger antiaging effects in aging mice, indicating that phosphorylation improved antiaging effect. [Display omitted] •Phosphorylated polysaccharide (PTPP-1) was prepared from polysaccharides from Trichosanthes peel (TPP-1).•TPP-1 had a significant antiaging activity on the D-galactose-induced aging mice.•The antiaging activity of PTPP-1 was more significant than that of TPP-1.

Trichosanthes anguina L. (family Cucurbitaceae) is a monoecious and diclinous plant that can be consumed as a vegetable and has anti-inflammatory and antioxidant effects. The chemical composition and content of volatile compounds in female and male buds of T. anguina were explored by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) technology combined with multivariate statistical analysis. The results showed that the content of the volatile compounds was different between female and male buds. 2,2,6-trimethyl-6-vinyltetrahydro-2H-pyran-3-ol and 2,2,6-trimethyl-6-vinyldihydro-2H-pyran-3(4H)-one were the main volatile compounds in both female and male buds. Based on the multivariate statistical analysis of orthogonal projections to latent structures discriminant analysis (OPLS-DA) and t-test, the content of seven compounds was significantly different between female and male buds. The content of three compounds in male buds was higher than that in female, i.e., (E)-4,8-dimethyl-1,3,7-nonatriene, 1,5,9,9-tetramethyl-1,4,7-cycloundecatriene, and (E)-caryophyllene. Conversely, the content of (Z)-4-hexen-1-ol, (Z)-3-hexenyl benzoate, (Z)-3-hexenyl salicylate, and 2-hexen-1-ol in female buds was higher than that in male buds. This is the first report on the difference in the volatile compounds between female and male buds of T. anguina, which enriches the basic research on the monoecious and diclinous plant and provides a reference for the study of plant sex differentiation.