Long-term ecological restoration increased plant diversity and soil total phosphorus content of the alpine flowing sand land in northwest Sichuan, China.

The ecological restoration techniques that combine grazing, sand barriers with willows, fertilization, artificial planting, and continuous management are increasingly adopted in the management of flowing sandy land in high-altitude and cold regions. However, few studies have focused on the long-term ecological restoration effects of such technologies. This study systematically compared the vegetation and soil characteristics under different ecological restoration durations (0 (CK), 3 (F1), 14 (F2), 26 (F3), and 46 (F4) years) in the alpine sandy land of northwest Sichuan. The results showed that, with the increase of ecological restoration durations, (1) the aboveground and underground biomass of plants, and species number significantly increased, while the shannon-wiener index, margalef index, and simpson index dramatically decreased; (2) in the early stage of ecological restoration (0-3 yr), Cyperaceae accounted for the main groups, while in the late stage of ecological restoration (14-46 yr), Leguminosae and Forb groups predominated; (3) ecological restoration durations significantly influenced the total phosphorus (TP) content at a soil depth of 0-60 cm, but soil organic carbon and C/P ratio were only significantly impacted at 40-60 cm; (4) the plant and soil characteristics of F1, F2, and F3 treatments were more similar, and CK and F4 treatments were clearly distinguished on PC1 of principal component analysis; (5) there was no significant correlation between Leguminosae groups and environmental factors. Instead, a correlation between total nitrogen (TN) and Forb groups, Gramineae groups, and Cyperaceae groups was revealed. TN was very significantly positively correlated with species diversity and TP. Long-term ecological restoration improved plants biomass, plant species diversity, functional plant groups, and increased soil TP content in the alpine sandy land of northwest Sichuan.

A comparative analysis of morphology, microstructure, and volatile metabolomics of leaves at varied developmental stages in Ainaxiang (Blumea balsamifera (Linn.) DC.).

Introduction: Ainaxiang (Blumea balsamifera (Linn.) DC.) is cultivated for the extraction of (-)-borneol and other pharmaceutical raw materials due to its abundant volatile oil. However, there is limited knowledge regarding the structural basis and composition of volatile oil accumulation in fresh B. balsamifera leaves. Methods: To address this problem, we compare the fresh leaves' morphology, microstructure, and volatile metabonomic at different development stages, orderly defined from the recently unfolded young stage (S1) to the senescent stage (S4). Results and discussion: Distinct differences were observed in the macro-appearance and microstructure at each stage, particularly in the B. balsamifera glandular trichomes (BbGTs) distribution. This specialized structure may be responsible for the accumulation of volatile matter. 213 metabolites were identified through metabolomic analysis, which exhibited spatiotemporal accumulation patterns among different stages. Notably, (-)-borneol was enriched at S1, while 10 key odor metabolites associated with the characteristic balsamic, borneol, fresh, and camphor aromas of B. balsamifera were enriched in S1 and S2. Ultra-microstructural examination revealed the involvement of chloroplasts, mitochondria, endoplasmic reticulum, and vacuoles in the synthesizing, transporting, and storing essential oils. These findings confirm that BbGTs serve as the secretory structures in B. balsamifera, with the population and morphology of BbGTs potentially serving as biomarkers for (-)-borneol accumulation. Overall, young B. balsamifera leaves with dense BbGTs represent a rich (-)-borneol source, while mesophyll cells contribute to volatile oil accumulation. These findings reveal the essential oil accumulation characteristics in B. balsamifera, providing a foundation for further understanding.

The recent progress in the research of extraction and functional applications of basil seed gum.

Basil seed gum (BSG) is a new hydrophilic colloid of natural plant origin. Extracted from basil seeds, it possesses excellent functional characteristics in terms of emulsification, rheology, gelation, stability, and adsorption, which are just as favorable as those of certain commercial gums. Besides, BSG has been widely used in food, medicine, industry, and many other fields for its physiological functions of weight reduction, detoxification, and control of blood sugar and cholesterol as a good dietary fiber. In this paper, we analyzed and discussed the extraction procedures, composition structures, functional characteristics, and modification strategies of BSG. In addition, we summarized the latest research on the applications of BSG in different industries to provide theoretical references for the high-value processing and utilization of BSG.

Transcriptome Analysis Reveals Genes Associated with Flooding Tolerance in Mulberry Plants.

Mulberry (Morus alba), a widely distributed economic plant, can withstand long-term flooding stress. However, the regulatory gene network underlying this tolerance is unknown. In the present study, mulberry plants were subjected to submergence stress. Subsequently, mulberry leaves were collected to perform quantitative reverse-transcription PCR (qRT-PCR) and transcriptome analysis. Genes encoding ascorbate peroxidase and glutathione S-transferase were significantly upregulated after submergence stress, indicating that they could protect the mulberry plant from flood damage by mediating ROS homeostasis. Genes that regulate starch and sucrose metabolism; genes encoding pyruvate kinase, alcohol dehydrogenase, and pyruvate decarboxylase (enzymes involved in glycolysis and ethanol fermentation); and genes encoding malate dehydrogenase and ATPase (enzymes involved in the TCA cycle) were also obviously upregulated. Hence, these genes likely played a key role in mitigating energy shortage during flooding stress. In addition, genes associated with ethylene, cytokinin, abscisic acid, and MAPK signaling; genes involved in phenylpropanoid biosynthesis; and transcription factor genes also showed upregulation under flooding stress in mulberry plants. These results provide further insights into the adaptation mechanisms and genetics of submergence tolerance in mulberry plants and could aid in the molecular breeding of these plants.

RNA sequencing in Artemisia annua L explored the genetic and metabolic responses to hardly soluble aluminum phosphate treatment.

Artemisia annua L. is a medicinal plant valued for its ability to produce artemisinin, a molecule used to treat malaria. Plant nutrients, especially phosphorus (P), can potentially influence plant biomass and secondary metabolite production. Our work aimed to explore the genetic and metabolic response of A. annua to hardly soluble aluminum phosphate (AlPO4, AlP), using soluble monopotassium phosphate (KH2PO4, KP) as a control. Liquid chromatography-mass spectrometry (LC-MS) was used to analyze artemisinin. RNA sequencing, gene ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were applied to analyze the differentially expressed genes (DEGs) under poor P conditions. Results showed a significant reduction in plant growth parameters, such as plant height, stem diameter, number of leaves, leaf areas, and total biomass of A. annua. Conversely, LC-MS analysis revealed a significant increase in artemisinin concentration under the AlP compared to the KP. Transcriptome analysis revealed 762 differentially expressed genes (DEGs) between the AlP and the KP. GH3, SAUR, CRE1, and PYL, all involved in plant hormone signal transduction, showed differential expression. Furthermore, despite the downregulation of HMGR in the artemisinin biosynthesis pathway, the majority of genes (ACAT, FPS, CYP71AV1, and ALDH1) were upregulated, resulting in increased artemisinin accumulation in the AlP. In addition, 12 transcription factors, including GATA and MYB, were upregulated in response to AlP, confirming their importance in regulating artemisinin biosynthesis. Overall, our findings could contribute to a better understanding the parallel transcriptional regulation of plant hormone transduction and artemisinin biosynthesis in A. annua L. in response to hardly soluble phosphorus fertilizer.

Identification of Key Aromatic Compounds in Basil (Ocimum L.) Using Sensory Evaluation, Metabolomics and Volatilomics Analysis.

Basil (Ocimum L.) is widely used as a flavor ingredient, however research on basil flavor is limited. In the current study, nine basil species were selected, including Ocimum basilicum L.var. pilosum (Willd.) Benth., Ocimum sanctum, Ocimum basilicum cinnamon, Ocimum gratissimum var. suave, Ocimum tashiroi, Ocimum basilicum, Ocimum americanum, Ocimum basilicum ct linalool, and Ocimum basilicum var. basilicum, and their fragrance and flavor characteristics were assessed by sensory evaluation. The results indicated that Ocimum basilicum var. basilicum and Ocimum gratissimum var. suave have a strong clove smell and exhibited a piquant taste. Metabolomics and volatilomics analyses measured 100 nonvolatile metabolites and 134 volatiles. Differential analysis showed that eugenol, γ-terpinene, germacrene D and malic acid were among the most varied metabolites in basil species. Combined with sensory evaluation results, correlation analysis revealed that ß-pinene and γ-cadinene contributed to the piquant smell, while eugenol and germacrene D contributed to the clove smell, and malic acid and L-(−)-arabitol contributed to the sweet flavor in basil. This study provided comprehensive flavor chemistry profiles of basil species and could be used as a guide for basil flavor improvement. The better understanding of objective sensory attributes and chemical composition of fresh basil could introduce the improved cultivars with preponderant traits, which is also in accordance with the various demands of breeders and growers, food producers, and consumers.

Potential distribution of Blumea balsamifera in China using MaxEnt and the ex situ conservation based on its effective components and fresh leaf yield.

Blumea balsamifera is a famous Chinese Minority Medicine, which has a long history in Miao, Li, Zhuang, and other minority areas. In recent years, due to the influence of natural and human factors, the distribution area of B. balsamifera resources has a decreasing trend. Therefore, it is very important to analyze the suitability of B. balsamifera in China. Following three climate change scenarios (SSP1-2.6, SSP2-4.5, and SSP5-8.5) under 2050s and 2070s, geographic information technology (GIS) and maximum entropy model (MaxEnt) were used to simulate the ecological suitability of B. balsamifera. The contents of L-borneol and total flavonoids of B. balsamifera in different populations were determined by gas chromatography (GC) and ultraviolet spectrophotometry (UV). The results showed that the key environmental variables affecting the distribution of B. balsamifera were mean temperature of coldest quarter (6.18-26.57 ℃), precipitation of driest quarter (22.46-169.7 mm), annual precipitation (518.36-1845.29 mm), and temperature seasonality (291.31-878.87). Under current climate situation, the highly suitable habitat was mainly located western Guangxi, southern Yunnan, most of Hainan, southwestern Guizhou, southwestern Guangdong, southeastern Fujian, and western Taiwan, with a total area of 24.1 × 104 km2. The areas of the moderately and poorly suitable habitats were 27.57 × 104 km2 and 42.43 × 104 km2, respectively. Under the future climate change scenarios, the areas of the highly, moderately, and poorly suitable habitats of B. balsamifera showed a significant increasing trend, the geometric center of the total suitable habitats of B. balsamifera would move to the northeast. In recent years, the planting area of B. balsamifera has been reduced on a large scale in Guizhou, and its ex situ protection is imperative. By comparison, the content of L-borneol, total flavonoids and fresh leaf yield had no significant difference between Guizhou and Hainan (P > 0.05), which indicated that Hainan is one of the best choice for ex situ protection of B. balsamifera.

New paraconic acids from the endophytic fungus Xylariaceae sp. J4.

Three new paraconic acids, xylariacinics A-C (1-3), were isolated from the endophyte Xylariaceae sp. J4 harbored in the medicinal plant Blumea balsamifera. Their structures were elucidated on the basis of extensive spectroscopic data including HRMS, and NMR. The antibacterial efficacies of compounds 1-3 were evaluated against a panel of bacteria such as Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. Their antifungal activities were also tested against Colletotrichum gloeosporioides. Unfortunately, all of them were inactive.

[Antibacterial secondary metabolites of Clonostachys rosea, an endophytic fungus from Blumea balsamifera (L.) DC.]

Endophytic fungus is an important treasure trove for discovery of structurally unusual and biologically diverse compounds. A phytochemical investigation on a fungus Clonostachys rosea inhabits inner tissue of Blumea balsamifera (L.) DC. was initiatedrecently in our lab. Six pure compounds were isolated through silica gel column chromatography, sephadex LH-20, and semi-preparative HPLC techniques, with bio-guided strategy. Their structures were characterized as verticillin A (1), (S)-(+)-fusarinolic acid (2), 8-hydroxyfusaric acid (3), cerebroside C (4), 3-Maleimide-5-oxime (5), and bionectriol A (6) by analyses of NMR and MS data. All compounds were tested in vitro antibacterial activities against four strains of bacteria, Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa, and results revealed that 1, 4 and 6 display notableinhibition againstthree bacteria, with MIC values ranging from 2 to 16 µg/mL. Our findings provide references for mining novel antibiotics from endophytes originated from Li Minority medicinal plant B. balsamifera (L.) DC.

Effects and Mechanisms of Total Flavonoids from Blumea balsamifera (L.) DC. on Skin Wound in Rats.

Chinese herbal medicine (CHM) evolved through thousands of years of practice and was popular not only among the Chinese population, but also most countries in the world. Blumea balsamifera (L.) DC. as a traditional treatment for wound healing in Li Nationality Medicine has a long history of nearly 2000 years. This study was to evaluate the effects of total flavonoids from Blumea balsamifera (L.) DC. on skin excisional wound on the back of Sprague-Dawley rats, reveal its chemical constitution, and postulate its action mechanism. The rats were divided into five groups and the model groups were treated with 30% glycerol, the positive control groups with Jing Wan Hong (JWH) ointment, and three treatment groups with high dose (2.52 g·kg-1), medium dose (1.26 g·kg-1), and low dose (0.63 g·kg-1) of total flavonoids from B. balsamifera. During 10 consecutive days of treatment, the therapeutic effects of rates were evaluated. On day 1, day 3, day 5, day 7, and day 10 after treatment, skin samples were taken from all the rats for further study. Significant increases of granulation tissue, fibroblast, and capillary vessel proliferation were observed at day 7 in the high dose and positive control groups, compared with the model group, with the method of 4% paraformaldehyde for histopathological examination and immunofluorescence staining. To reveal the action mechanisms of total flavonoids on wound healing, the levels of CD68, vascular endothelial growth factor (VEGF), transforming growth factor-ß1 (TGF-ß1), and hydroxyproline were measured at different days. Results showed that total flavonoids had significant effects on rat skin excisional wound healing compared with controls, especially high dose ones (p < 0.05). Furthermore, the total flavonoid extract was investigated phytochemically, and twenty-seven compounds were identified from the total flavonoid sample by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry/diode array detector (UPLC-Q-TOF-MS/DAD), including 16 flavonoid aglucons, five flavonoid glycosides (main peaks in chromatogram), five chlorogenic acid analogs, and 1 coumarin. Reports show that flavonoid glycoside possesses therapeutic effects of curing wounds by inducing neovascularization, and chlorogenic acid also has anti-inflammatory and wound healing activities; we postulated that all the ingredients in total flavonoids sample maybe exert a synergetic effect on wound curing. Accompanied with detection of four growth factors, the upregulation of these key growth factors may be the mechanism of therapeutic activities of total flavonoids. The present study confirmed undoubtedly that flavonoids were the main active constituents that contribute to excisional wound healing, and suggested its action mechanism of improving expression levels of growth factors at different healing phases.

[Analysis of metabolic pathway of terpenoids in Blumea balsamifera].

In order to provide a theoretical basis for the regulation of active ingredient, the terpenoids metabolic pathway and specific enzymes in Blumea balsamifera are investigated. Basing on transcriptome information, B. balsamifera terpenoids metabolic pathway was analyzed in KEGG data base. Four metabolic pathway of terpenoids were found in KEGG data base. They were terpenoid backbone biosynthesis, monoterpenoid biosynthesis, diterpenoid biosynthesis, sesquiterpenoid and triterpenoid biosynthesis, contained 103, 10, 29,59 genes, respectively. Through the analysis of the enzyme and product in the pathway, the result showed that there were 8 kinds of monoterpenes, 3 kinds of diterpenes, 3 kinds of triterpenes and sesquiterpenes. The mainly key enzymes were deoxyxylulose 5-phosphate synthase, HMG-CoA reductase and allyl transferase system. In B. balsamifera, there were relatively few monoterpenes synthetic enzymes, while the type of products was much more than other terpenes. This may be relate to the non-specific catalytic characteristic of monoterpene synthase. It is expected to improve the yield of terpenoids in B. balsamifera by analysis the pathways and regulation the key enzymes.

[Metabolic pathway of flavonoids in Blumea balsamifera].

To better understand the biosynthetic pathway of flavonoids in Blumea balsamifera, and to increase the biosynthesis of flavone and flavonol amount, we carried out this research. Basing on transcriptome information, B. balsamifera flavonoids metabolic pathway was analyzed in KEGG data base. The result demonstrated that two metabolic pathways of flavonoids were found in KEGG data base. They were flavonoid biosynthesis pathway (No.ko00941) that contained 32 genes and flavone and flavonol biosynthesis pathway (No.ko00944) that contained 12 genes. Metabolic pathway of flavonoids in B. balsamifera was similar to that in other plants. Chalcone synthase (CHS) and Chalconeisomerase were much likely to be key enzymes on metabolic pathway of flavonoids in B. balsamifera. HCT was much relevant to biosynthesis of favones.

[Phylogeny and functional divergence of ω-6 and ω-3 fatty acid desaturase families].

The deduced amino acid sequences characteristics, phylogeny, and functional diverge of ω-6 and ω-3 fatty acid desaturase families were analyzed by using Bioinformatics methods. The results showed that all the deduced amino acid sequences shared three highly conserved histidine rich motifs (Hisbox). All the plastidial ω-6 and ω-3 fatty acid desaturases possessed putative N-terminal signal peptide with different amino acids. A relatively conserved hydrophobic region composed of 10 amino-acid residues was found in the middle of signal peptides, which is presumed to be the functional region of the signal peptide of these enzymes. Most of the plant microsomal ω-6 and ω-3 fatty acid desaturases (FAD2 and FAD3) contained a KKXX-like motif of endoplasmic reticulum (ER) retention signal at the C-terminus. However, no such motif was detected in safflower CtFAD2-3, CtFAD2-4, CtFAD2-5, CtFAD2-6, and CtFAD2-7, while an aromatic aa enriched signal (YKNK) was found at their C-terminus which has been reported to be both necessary and sufficient for maintaining localization of the enzymes in the ER. All the amino acid sequences were divided into four categories through phylogenetic analysis. It was suggested that ω-3 fatty acid desaturase originates in a prokaryotic lineage from ω-6 fatty acid desaturase. Both plastidial and microsomal ω-3 fatty acid desaturases could be divided into dicotyledonous and monocotyledonous subgroups, which inferred that functional differentiation of plastidial and microsomal ω-3 fatty acid desaturases had been formed before the divergence of dicotyledonous and monocotyledonous plants. Seed type and housekeeping type FAD2 diverged after the formation of dicotyledonous plants. Except for plant FAD3/plant FAD2, posterior probability values over 0.80 amino acid sites were identified among the functional differentiation subsets, which were mainly distributed at the front and back end of Hisbox I, and the front end of Hisbox II. This indicated that the variations of these amino acid sites played an important role in the size and conformation of protein functional domains and subfamily functional divergence.

[Effect of different fertilizing levels on yields and efficiency components of Pinellia ternata from Sichuan].

OBJECTIVE: The effect of the different fertilizing levels on yields and main active components of Pinellia ternata from Sichuan was studied under the cultivated condition, in order to provide theoretical basis for the standardized cultivation. METHOD: Using one of the wild populations of P. ternate from Sichuan as tested material, the experiment was performed with orthogonal designing methods L25 (5(3)). During growth and development period, agronomic traits such as number of sprouting, inflorescence and bulblets were counted. After harvesting, main chemical compositions, growth and proliferation rates were determined. RESULT: In different fertilizing levels, the P. ternata from Sichuan showed the same growth rhythm, though there were significant difference (P < 0.05) among the average emergence rate,whereas extremely significant difference (P < 0.01) were detected among the average ratio of bolting, the average bulbils, individual growth rate, individual proliferation rate, beta-sitosterol and alkaloid content under different fertilizing levels. Nitrogenous fertilizer that affected the content of alkaloids and beta-sitosterol were extremely significant (P < 0.01), whereas phosphate and potassium fertilizer had no significant effect. The effect of fertilizer factor and inter effects on beta-sitosterol showed no significant effect, but have influence on other indexes. CONCLUSION: The optimum fertilizer composition was 315 kg x hm(-2) of nitrogen, 225 kg x hm(-2) of P2O5 and 270 kg x hm(-2) of K2O.