Integrated metabolome and transcriptome analysis unveils novel pathway involved in the fruit coloration of Nitraria tangutorum Bobr.

BACKGROUND: The desert shrub Nitraria tangutorum Bobr. is important for its resistance to salt and alkali in Northwest China. It is an ecologically important species in this region and provides edible and medicinal berries. This study showed a mutant of N. tangutorum (named Jincan, JC) that has a strong yellow pericarp vs red in a wild type (represented by NT). RESULTS: In this study, the secondary metabolic and molecular mechanisms responsible for Nitraria fruit coloration were investigated using LC-MS-based widely targeted metabolomics and transcriptomics data. As a result of our study, 122 and 104 flavonoid metabolites were differentially expressed throughout the mature and transition stages between JC and NT, respectively. Furthermore, two cyanidin derivatives (cyanidin 3-O-glucoside and cyanidin-3-O-(2''-O-glucosyl) glucoside) and one pelargonidin derivative (pelargonidin-3-O-glucoside) were identified only in the NT phenotype. The functional genes F3H (flavanone 3-hydroxylase), F3'H (flavonoid 3'-hydroxylase) and UFGT (flavonoid 3-O-glucosyltransferase) and the transcription factors MYB, bHLH, NAC and bZIP were significantly downregulated in JC. Meanwhile, the activity of UFGT was extremely low in both periods of JC, with a five-fold higher enzymatic activity of UFGT in RT than in YT. In summary, due to the lack of catalysis of UGFT, yellow fruit of JC could not accumulate sufficient cyanidin and pelargonidin derivatives during fruit ripening. CONCLUSION: Taken together, our data provide insights into the mechanism for the regulation of anthocyanin synthesis and N. tangutorum fruit coloration and provide a theoretical basis to develop new strategies for developing bioactive compounds from N. tangutorum fruits.

CIPK11: a calcineurin B-like protein-interacting protein kinase from Nitraria tangutorum, confers tolerance to salt and drought in Arabidopsis.

BACKGROUND: The CIPKs are a group of plant-specific Ser/Thr protein kinases acting in response to calcium signaling, which plays an important role in the physiological and developmental adaptation of plants to adverse environments. However, the functions of halophyte-derived CIPKs are still poorly understood, that limits a potential application of CIPKs from halophytes for improving the tolerance of glycophytes to abiotic stresses. RESULTS: In this study, we characterized the NtCIPK11 gene from the halophyte Nitraria tangutorum and subsequently analyzed its role in salt and drought stress tolerance, using Arabidopsis as a transgenic model system. NtCIPK11 expression was upregulated in N. tangutorum root, stem and blade tissues after salt or drought treatment. Overexpressing NtCIPK11 in Arabidopsis improved seed germination on medium containing different levels of NaCl. Moreover, the transgenic plants grew more vigorously under salt stress and developed longer roots under salt or drought conditions than the WT plants. Furthermore, NtCIPK11 overexpression altered the transcription of genes encoding key enzymes involved in proline metabolism in Arabidopsis exposed to salinity, however, which genes showed a relatively weak expression in the transgenic Arabidopsis undergoing mannitol treatment, a situation that mimics drought stress. Besides, the proline significantly accumulated in NtCIPK11-overexpressing plants compared with WT under NaCl treatment, but that was not observed in the transgenic plants under drought stress caused by mannitol application. CONCLUSIONS: We conclude that NtCIPK11 promotes plant growth and mitigates damage associated with salt stress by regulating the expression of genes controlling proline accumulation. These results extend our understanding on the function of halophyte-derived CIPK genes and suggest that NtCIPK11 can serve as a candidate gene for improving the salt and drought tolerance of glycophytes through genetic engineering.

A rapid high-performance liquid chromatography separation of a new anthocyanin from Nitraria tangutorum.

In this work, a rapid high-performance liquid chromatography method was developed to efficiently purify anthocyanin from Nitraria tangutorum based on reversed-phase column. A new anthocyanin was purified from N. tangutorum and elucidated on the basis of extensive spectroscopic analysis, including one- and two-dimensional nuclear magnetic resonance, as well as high-resolution mass spectrometry (HR-MS) data. The new anthocyanin was elucidated as cyanidin 3-[2″-(6‴-coumaroyl)-glucosyl]-glucoside.

Rapid Determination of Amino Acids of Nitraria tangutorum Bobr. from the Qinghai-Tibet Plateau Using HPLC-FLD-MS/MS and a Highly Selective and Sensitive Pre-Column Derivatization Method.

Amino acids are indispensable components of living organisms. The high amino acid content in Nitraria tangutorum Bobr. fruit distinguishes it from other berry plants and is of great significance to its nutritional value. Herein, using 10-ethyl-acridine-3-sulfonyl chloride as a fluorescent pre-column labeling reagent, a method for the efficient and rapid determination of amino acid content in N. tangutorum by pre-column fluorescence derivatization and on-line mass spectrometry was established and further validated. The limits of detection (signal-to-noise ratio = 3) were between 0.13 and 1.13 nmol/L, with a linear coefficient greater than 0.997 and a relative standard deviation between 1.37% and 2.64%. In addition, the method required a short analysis time, separating 19 amino acids within 20 min. Subsequently, the method was used to analyze the amino acid content of Nitraria tangutorum Bobr. from tissues retrieved from seven regions of the Qinghai-Tibet Plateau. Nitraria tangutorum Bobr. was shown to contain a large amount of amino acids, with the total content and main amino acid varying between the different tissues. This research supports the nutritional evaluation, quality control, and development and utilization of Nitraria tangutorum Bobr.

Gene transcript profiles in the desert plant Nitraria tangutorum during fruit development and ripening.

Nitraria tangutorum Bobr., a valuable wild shrub distributed in Northwest China, produces edible and medicinal berries. However, little is known about the molecular mechanisms of its fruit development and ripening. We performed de novo transcriptome sequencing of N. tangutorum fruit using the Illumina HiSeq™ 2000 sequencing platform. More than 62.94 million reads were obtained and assembled into 69,306 unigenes (average length, 587 bp). These unigenes were annotated by querying against five databases (Nr, Swiss-Prot, GO, COG, and KEGG); 42,929 and 26,809 unigenes were found in the Nr and Swiss-Prot databases, respectively. In ortholog analyses, 33,363 unigenes were assigned with one or more GO terms, 15,537 hits were aligned to 25 COG classes, and 24,592 unigenes were classified into 128 KEGG pathways. Digital gene expression analyses were conducted on N. tangutorum fruit at the green (S1), yellow (S2), and red (S3) developmental stages. In total, 8240, 5985, and 4994 differentially expressed genes (DEGs) were detected for S1 vs. S2, S1 vs. S3, and S2 vs. S3, respectively. Cluster analyses showed that a large proportion of DEGs related to plant hormones and transcription factors (TFs) showed high expression in S1, down-regulated expression in S2, and up-regulated expression in S3. We analyzed the expression patterns of 23 genes encoding 12 putative enzymes involved in flavonoid biosynthesis. The expression profiles of 10 DEGs involved in flavonoid biosynthesis were validated by Q-PCR analysis. The assembled and annotated transcriptome sequences and gene expression profile analyses provide valuable genetic resources for research on N. tangutorum.

Demographic patterns of two related desert shrubs with overlapping distributions in response to past climate changes.

Numerous studies have revealed that past geological events and climatic fluctuations had profoundly affected the genetic structure and demographic patterns of species. However, related species with overlapping ranges may have responded to such environmental changes in different ways. In this study, we compared the genetic structure and population dynamics of two typical desert shrubs with overlapping distributions in northern China, Nitraria tangutorum and Nitraria sphaerocarpa, based on chloroplast DNA (cpDNA) variations and species distribution models. We sequenced two cpDNA fragments (trnH-trnA and atpH-atpI) in 633 individuals sampled from 52 natural populations. Twenty-four chlorotypes, including eight rare chlorotypes, were identified, and a single dominant haplotype (H4) widely occurred in the entire geographical ranges of the two species. There were also a few distinctive chlorotypes fixed in different geographical regions. Population structure analyses suggested that the two species had significantly different levels of total genetic diversity and interpopulation differentiation, which was highly likely correlated with the special habitat preferences of the two species. A clear phylogeographic structure was identified to exist among populations of N. sphaerocarpa, but not exist for N. tangutorum. The neutral tests, together with the distribution of pairwise differences revealed that N. tangutorum experienced a sudden demographic expansion, and its expansion approximately occurred between 21 and 7 Kya before present, while a rapid range expansion was not identified for N. sphaerocarpa. The ecological niche modeling (ENM) analysis indicated that the potential ranges of two species apparently fluctuated during the past and present periods, with obvious contraction in the Last Glacial Maximum (LGM) and recolonization in the present, respectively, comparing to the Last Interglacial (LIG). These findings suggest that the two species extensively occurred in the Northwest of China before the Quaternary, and the current populations of them originated from a few separated glacial refugia following their habitat fragmentation in the Quarternary. Our results provide new insights on the impact of past geological and climatic fluctuations on the population dynamics of desert plants in northwestern China, and further enforce the hypothesis that there were several independent glacial refugia for these species during the Quaternary glaciations.

Evolution stage identification and spatial pattern analysis of Nitraria tangutorum nebkhas based on Gaofen-2 satellite data.

The quantitative extraction and evolution stage identification of the Nitraria tangutorum nebkhas are the basis for the restoration of regional plants and the reconstruction of degraded ecosystems. In this paper, the Nitraria tangutorum nebkha in Dengkou County of China was taken as the research object. Through the spectral and texture information of Gaofen-2 satellite image, the quantitative extraction of Nitraria tangutorum nebkha area and coverage information was completed using methods of gray threshold method, mathematical morphology, FCLSU mixed pixel decomposition, kernel density spatial analysis; the current evolution stage of the Nitraria tangutorum nebkha was identified, and their spatial distribution characteristics were analyzed. The results showed that: (1) The user accuracy and mapping accuracy of Nitraria tangutorum nebkha extracted from Random Forest combined with object-oriented classification method were up to 90.32%. (2) The method proposed can achieve an accuracy of 93.76% in extracting the spatial position of Nitraria tangutorum nebkhas. (3) The evolution of Nitraria tangutorum nebkhas can be divided into three stages: embryonic or developmental stage, stable stage, and declining stage, with a proportion of 60.70%, 20.97%, and 18.33%, respectively; The Nitraria tangutorum nebkhas in the study area is mainly in their embryonic or developmental stage, and the proportion of Nitraria tangutorum nebkhas in the declining stage is also large. It can provide technical and theoretical support for the precise extraction of nebkhas in arid and semi-arid desert areas, the identification of their current evolutionary stages, and the study of their spatial distribution patterns.

A systematic review on the morphology structure, propagation characteristics, resistance physiology and exploitation and utilization of Nitraria tangutorum Bobrov.

Nitraria tangutorum Bobrov., belonging to the family Nitrariaceae, is a drought-tolerant and salt-loving plant and has drawn attention for its good economic and ecological value. As one of the main group species and dominant species in China's desert and semi-desert regions, N. tangutorum possesses superior tolerance to drought, high temperature, cold, barren, high salinity and alkalinity and wind and sand. Its root system is well developed, with many branches and a strong germination capacity. Once buried in sandy soil, N. tangutorum can quickly produce a large number of adventitious roots, forming new plants and continuously expanding the shrubs, forming fixed and semi-fixed shrub sand dunes. Sand dune shrubs can trap and fix a large amounts of quicksand, prevent desert expansion and erosion, and play an important role in maintaining regional ecosystem balance and improving ecological environmental quality. In addition, the phytochemical screening studies report that N. tangutorum contains an abundance of various compounds including flavonoids, alkaloids, phenolic acids and polysaccharides. These compounds confer a range of beneficial bioactivities such as antioxidant, anti-inflammatory, anti-tumor, anti-fatigue, liver protection, neuroprotection, cardiovascular protection, lowering blood lipid, regulating blood sugar level and immunoregulation. The fruits of N. tangutorum also contain vitamin C, amino acids, minerals and microelements. It has been traditionally used as a nutritional food source and in folk medicine to treat diseases of the spleen and stomach, abnormal menstruation, indigestion, and hyperlipidemia. N. tangutorum, as a wild plant with medicinal and edible homology, possesses remarkable economic and medicinal values. This detailed, comprehensive review gathers and presents all the information related to the morphological structure, propagation characteristics, resistance physiology and exploitation and utilization of N. tangutorum, providing a theoretical basis for the researchers to conduct future in-depth research on N. tangutorum.

Transcriptome Profiling and Chlorophyll Metabolic Pathway Analysis Reveal the Response of Nitraria tangutorum to Increased Nitrogen.

To identify genes that respond to increased nitrogen and assess the involvement of the chlorophyll metabolic pathway and associated regulatory mechanisms in these responses, Nitraria tangutorum seedlings were subjected to four nitrogen concentrations (N0, N6, N36, and N60: 0, 6, 36, and 60 mmol·L-1 nitrogen, respectively). The N. tangutorum seedling leaf transcriptome was analyzed by high-throughput sequencing (Illumina HiSeq 4000), and 332,420 transcripts and 276,423 unigenes were identified. The numbers of differentially expressed genes (DEGs) were 4052 in N0 vs. N6, 6181 in N0 vs. N36, and 3937 in N0 vs. N60. Comparing N0 and N6, N0 and N36, and N0 and N60, we found 1101, 2222, and 1234 annotated DEGs in 113, 121, and 114 metabolic pathways, respectively, classified in the Kyoto Encyclopedia of Genes and Genomes database. Metabolic pathways with considerable accumulation were involved mainly in anthocyanin biosynthesis, carotenoid biosynthesis, porphyrin and chlorophyll metabolism, flavonoid biosynthesis, and amino acid metabolism. N36 increased δ-amino levulinic acid synthesis and upregulated expression of the magnesium chelatase H subunit, which promoted chlorophyll a synthesis. Hence, N36 stimulated chlorophyll synthesis rather than heme synthesis. These findings enrich our understanding of the N. tangutorum transcriptome and help us to research desert xerophytes' responses to increased nitrogen in the future.

Exogenous abscisic acid and sodium nitroprusside regulate flavonoid biosynthesis and photosynthesis of Nitraria tangutorum Bobr in alkali stress.

Abscisic acid (ABA) and nitric oxide (NO) are involved in mediating abiotic stress-induced plant physiological responses. Nitraria tangutorum Bobr is a typical salinized desert plant growing in an arid environment. In this study, we investigated the effects of ABA and NO on N.tangutorum seedlings under alkaline stress. Alkali stress treatment caused cell membrane damage, increased electrolyte leakage, and induced higher production of reactive oxygen species (ROS), which caused growth inhibition and oxidative stress in N.tangutorum seedlings. Exogenous application of ABA (15µm) and Sodium nitroprusside (50µm) significantly increased the plant height, fresh weight, relative water content, and degree of succulency in N.tangutorum seedlings under alkali stress. Meanwhile, the contents of ABA and NO in plant leaves were significantly increased. ABA and SNP can promote stomatal closure, decrease the water loss rate, increase leaf surface temperature and the contents of osmotic regulator proline, soluble protein, and betaine under alkali stress. Meanwhile, SNP more significantly promoted the accumulation of chlorophyll a/b and carotenoids, increased quantum yield of photosystem II (φPSII) and electron transport rate (ETRII) than ABA, and decreased photochemical quenching (qP), which improved photosynthetic efficiency and accelerated the accumulation of soluble sugar, glucose, fructose, sucrose, starch, and total sugar. However, compared with exogenous application of SNP in the alkaline stress, ABA significantly promoted the transcription of NtFLS/NtF3H/NtF3H/NtANR genes and the accumulation of naringin, quercetin, isorhamnetin, kaempferol, and catechin in the synthesis pathway of flavonoid metabolites, and isorhamnetin content was the highest. These results indicate that both ABA and SNP can reduce the growth inhibition and physiological damage caused by alkali stress. Among them, SNP has a better effect on the improvement of photosynthetic efficiency and the regulation of carbohydrate accumulation than ABA, while ABA has a more significant effect on the regulation of flavonoid and anthocyanin secondary metabolite accumulation. Exogenous application of ABA and SNP also improved the antioxidant capacity and the ability to maintain Na+/K+ balance of N. tangutorum seedlings under alkali stress. These results demonstrate the beneficial effects of ABA and NO as stress hormones and signaling molecules that positively regulate the defensive response of N. tangutorum to alkaline stress.

Allometry of bud dynamic pattern and linkage between bud traits and ecological stoichiometry of Nitraria tangutorum under fertilizer addition.

Affected by the pressure and constraints of available resources, plant growth and development, as well as plant life history strategies, usually vary with environmental conditions. Plant buds play a crucial role in the life history of woody plants. Nitraria tangutorum is a common dominant woody species in desertified areas of northern China and its growth is critical to the desert ecosystem. Revealing the allometry of N. tangutorum aboveground bud fates and the linkage between bud traits and plant nutrient contents and stoichiometric ratios can be useful in understanding plant adaptation strategy. We applied seven nitrogen and phosphorus fertilizer addition treatments to natural N. tangutorum ramets in Ulan Buh Desert in three consecutive years. We surveyed three types of aboveground buds (dormant buds, vegetative buds, and reproductive buds) in each N. tangutorum ramet, then measured the plant carbon (C), nitrogen (N), and phosphorus (P) contents and ratios during three consecutive years. We specified that reserve growth potential (RGP), vegetative growth intensity (VGI) and sexual reproduction effort (SRE) are the three indices of bud dynamic pattern. The results showed that the bud dynamic pattern of N. tangutorum ramets differed significantly among different fertilizer addition treatments and sampling years. The allometry of RGP, VGI, and SRE was obvious, showing size dependence. The allometric growth relationship fluctuated among the sampling years. The linkage between bud traits and plant stoichiometric characteristics of N. tangutorum ramets showed close correlation with plant P content, C:P and N:P ratios, no significant correlation with plant C content, N content and C:N ratio. These results contribute to an improved understanding of the adaptive strategies of woody plants growing in desert ecosystems and provide insights for adoption of effective measures to restore and conserve plant communities in arid and semi-arid regions.

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As a special bio-geomorphic landscape in the Qaidam desert area, Nitraria tangutorum nebkhas play a critical role in fixing quicksand, improving soil quality, and maintaining the stability of regional ecological environment. Taking the N. tangutorum nebkhas with coverage of approximately 15%, 25%, 45% and 60% in Gahai Lake area of Qaidam Basin as the research objects, we analyzed the vertical distribution and enrichment characteristics of soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), total potassium (TK), alkali-hydrolyzable nitrogen (AN), available phosphorus (AP) and available potassium (AK). The results showed that the contents of SOM, TN, TP, TK, AN, AP and AK varied in the range of 1.67-10.22 g·kg-1, 0.05-0.42 g·kg-1, 0.31-0.54 g·kg-1, 15.87-18.84 g·kg-1, 2.26-11.68 mg·kg-1, 0.80-15.00 mg·kg-1 and 45-161 mg·kg-1, respectively. Vertically, soil nutrients in the N. tangutorum nebkhas with 15% coverage showed a decreasing trend first then increased, and then decreased again with the increase of soil depth, except for TP, which did not show any significant change. In the nebkhas with 25%, 45% and 60% coverage, SOM, TN, AN, TP and AP all showed a decreasing trend with increasing soil depth ,whereas TK and AK did not change significantly with soil layer. Above the nebkhas ground level of N. tangutorum, SOM, TN, TK, AN, AP and AK were all enriched, especially in the surface layer.Aamong all the nutrients, the enrichment rate of AN reached 5.19. In addition, below the nebkhas ground level of N. tangutorum, TN, AN, TK, AK and AP also showed enrichment. SOM, TN, AN, TP, AP, TK and AK were all significantly positively correlated with soil water content, and negatively correlated with altitude. All nutrients except TP were mainly affected by altitude. In conclusion, soil nutrient content of N. tangutorum nebkhas was the highest in the surface layer, the enrichment effect of which was not only reflected in the interior of the nebkhas, but also below the ground level of the nebkhas. Our results could provide reference for the scientific utilization of N. tangutorum nebkhas and ecological environment protection in Qaidam Basin area.

Root exudates and rhizosphere soil bacterial relationships of Nitraria tangutorum are linked to k-strategists bacterial community under salt stress.

When plants are subjected to various biotic and abiotic stresses, the root system responds actively by secreting different types and amounts of bioactive compounds, while affects the structure of rhizosphere soil bacterial community. Therefore, understanding plant-soil-microbial interactions, especially the strength of microbial interactions, mediated by root exudates is essential. A short-term experiment was conducted under drought and salt stress to investigate the interaction between root exudates and Nitraria tangutorum rhizosphere bacterial communities. We found that drought and salt stress increased rhizosphere soil pH (9.32 and 20.6%) and electrical conductivity (1.38 and 11 times), respectively, while decreased organic matter (27.48 and 31.38%), total carbon (34.55 and 29.95%), and total phosphorus (20 and 28.57%) content of N. tangutorum rhizosphere soil. Organic acids, growth hormones, and sugars were the main differential metabolites of N. tangutorum under drought and salt stress. Salt stress further changed the N. tangutorum rhizosphere soil bacterial community structure, markedly decreasing the relative abundance of Bacteroidota as r-strategist while increasing that of Alphaproteobacteria as k-strategists. The co-occurrence network analysis showed that drought and salt stress reduced the connectivity and complexity of the rhizosphere bacterial network. Soil physicochemical properties and root exudates in combination with salt stress affect bacterial strategies and interactions. Our study revealed the mechanism of plant-soil-microbial interactions under the influence of root exudates and provided new insights into the responses of bacterial communities to stressful environments.

Nitric oxide alleviates salt-induced stress damage by regulating the ascorbate-glutathione cycle and Na+/K+ homeostasis in Nitraria tangutorum Bobr.

Nitric oxide (NO) is an important signaling molecule involved in mediation of salt stress induced physiological responses in plants. In this study, we investigated the effect of NO on Nitraria tangutorum seedlings exposed to salt stress. Exogenous application of NO donor, sodium nitroprusside (SNP) increased fresh weight, shoot and root elongation and decreased electrolyte leakage and malondialdehyde (MDA) content in N. tangutorum seedlings under salt stress. Simultaneously, leaf senescence and root damage induced by salt stress were alleviated. SNP effectively increased NO content both in leaves and roots of plants under salt stress. Meanwhile, SNP activated the ascorbate-glutathione (AsA-GSH) cycle by increasing antioxidants contents, antioxidant enzymes activities, and related genes expression, thereby scavenging reactive oxygen species (ROS) and alleviating oxidative damage caused by salt stress. SNP alleviated salt stress induced ion toxicity by promoting Na+ efflux and ion transporter gene expression and reducing Na+ content and the Na+/K+ ratio. In addition, application of NO specific scavenger cPTIO and mammalian NO synthase inhibitor L-NAME sifnificantly aggravated stress damage in plant under salt stress. These results show the beneficial impacts of NO as a stress-signaling molecule that positively regulates defense response in N. tangutorum to salt stress.

Effect of Plant Growth Regulators on Osmotic Regulatory Substances and Antioxidant Enzyme Activity of Nitraria tangutorum.

Plant growth regulators (PGRs) are natural hormones and synthetic hormone analogues. At low concentrations, PGRs have the ability to influence cell division, cell expansion, and cell structure and function, in addition to mediating environmental stress. In this study, experiments were conducted to determine how exogenous PGRs indole acetic acid (IAA), abscisic acid (ABA), and gibberellic acid (GA) influenced osmotic regulatory substances and activity of antioxidant enzymes in Nitraria tangutorum. Using a completely randomized design, IAA, ABA, and GA3 were applied as foliar spray at concentrations of 50 mg/L, 100 mg/L, 150 mg/L, and 200 mg/L to N. tangutorum shrubs. Some selected shrubs did not receive any treatment and served as the control (Ck). The results showed that the foliar spray of IAA, ABA, and GA3 significantly increased the content of osmotic regulatory substances (soluble sugar, soluble protein, and proline) and antioxidant enzymes (SOD and POD) at most concentrations. In addition, the malondialdehyde (MDA) content significantly reduced after treatment, but after regrowth of coppiced shrubs, lipid peroxidation increased and was still lower than Ck. Our study provides evidence that 100 mg/L 150 mg/L, and 200 mg/L concentrations of IAA, ABA, and GA3 treatments are effective for enhancing osmotic regulatory substances and the activity of antioxidant enzymes in N. tangutorum, which offers an effective strategy not only for increasing tolerance to abiotic and biotic stresses, but also improving the adaptability of N. tangutorum shrubs to the environment.

Pollen Morphology of Some Species from Genus Nitraria.

An analysis of pollen grains (in Nitraria sibirica Pall., N. schoberi L., N. komarovii Iljin & Lava ex Bobrov, and N. pamirica L. Vassil.) was performed on natural material collected in Russia, Kazakhstan, and Tajikistan. Herbarium specimens from the collection at Komarov Botanical Institute, Russian Academy of Sciences (N. tangutorum Bobrov and N. praevisa Bobrov) were examined, too. Pollen grains of two species-N. pamirica and N. praevisa-were studied for the first time. N. tangutorum and N. praevisa were found to have the perprolate pollen shape, whereas N. pamirica was found to have the subprolate shape. An intraspecific differentiation of N. sibirica was noted. Populations of N. sibirica (Taskarasu, Karatal, and Basshi) possess pollen grains of the subprolate or prolate shape, striate and perforate exine ornamentation, and a longer equatorial axis and a shorter polar axis than other specimens of N. sibirica. N. schoberi in all populations had anomalous shapes of some pollen grains. Overall, we demonstrated that the length ratio of the polar axis to the equatorial axis, characteristics of pollen in polar view, colpus morphology, and surface ornamentation of pollen grains in the genus Nitraria are of great taxonomic importance for the identification of species.

The Full-Length Transcriptome Sequencing and Identification of Na+/H+ Antiporter Genes in Halophyte Nitraria tangutorum Bobrov.

Nitraria tangutorum Bobrov is a halophyte that is resistant to salt and alkali and is widely distributed in northwestern China. However, its genome has not been sequenced, thereby limiting studies on this particular species. For species without a reference genome, the full-length transcriptome is a convenient and rapid way to obtain reference gene information. To better study N. tangutorum, we used PacBio single-molecule real-time technology to perform full-length transcriptome analysis of this halophyte. In this study, a total of 21.83 Gb of data were obtained, and 198,300 transcripts, 51,875 SSRs (simple sequence repeats), 55,574 CDS (coding sequence), and 74,913 lncRNAs (long non-coding RNA) were identified. In addition, using this full-length transcriptome, we identified the key Na+/H+ antiporter (NHX) genes that maintain ion balance in plants and found that these are induced to express under salt stress. The results indicate that the full-length transcriptome of N. tangutorum can be used as a database and be utilized in elucidating the salt tolerance mechanism of N. tangutorum.

A new flavonol acylglycoside from the fruits of Nitraria tangutorum Bobr.

Phytochemical investigation on the ethanol extract of the fruits of Nitraria tangutorum Bobr. led to the isolation of a new flavonol acylglycoside tangutoside (7) and six known ones including p-formylphenol (1), 3-formylindole (2), rutin (3), prionitisid B (4), quercitrin (5) and flazin (6). Among them, compounds 2, 4, and 5 were isolated from Nitraria tangutorum Bobr. for the first time. Their structures were identified by spectroscopic methods and by comparison with data reported in the references. The compounds 3, 4, 5, and 7 were demonstrated to possess strong DPPH radical scavenging ability with IC50 values of 31.45 µg/mL, 19.21 µg/mL, 59.30 µg/mL, and 44.54 µg/mL, respectively.

Characterization, antioxidant, and neuroprotective effects of anthocyanins from Nitraria tangutorum Bobr. fruit.

An anthocyanin-rich extract was obtained from Nitraria tangutorum Bobr. fruit, namely ANF, and its composition, antioxidant and neuroprotective effects were studied. Nine anthocyanins were identified from the ANF using UPLC-Triple-TOF/MS analysis, and cyanidin-3-[2''-(6'''-coumaroyl)-glucosyl]-glucoside (C3G) is the most abundant anthocyanin (87.06%). ANF exhibited high ferric reducing antioxidant power (FRAP) and ABTS radical scavenging activity. The online HPLC-DPPH screening revealed that C3G contributed the highest antioxidant capacity. ANF showed potential neuroprotective effects by relieving d-Galactose-induced memory deficits, reducing overexpression of receptor for advanced glycation end products (RAGE) and amyloid-beta42 (Aß42) in the hippocampus of rats. Besides, ANF could inhibit oxidative stress by reducing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in the hippocampus, while elevating amounts of total superoxide dismutase (T-SOD) and glutathione (GSH) in the serum of rats. Thus, ANF has great potential in the development of food and health products related to antioxidant and neuroprotective effects.

Nitraria tangutorum Bobr.-derived polysaccharides protect against LPS-induced lung injury.

Nitraria tangutorum Bobr. is suggested to be active in immunoregulation and antioxidation. However, the in vivo bioactivity of N. tangutorum Bobr.-derived polysaccharides (NTP) and their anti-inflammatory activity have not been addressed. In the present study, we extracted and purified polysaccharides from N. tangutorum Bobr. and determined their anti-inflammatory activities in vivo. HPGPC, UHPLC/DAD, and NMR analyses identified that the monosaccharide components of NTP were Man, Rha, GalUA, Glu, Gal, and Ara, with relative contents of 3.52%, 15.08%, 10.00%, 26.73%, 38.08%, and 6.59%, respectively. In mice with lipopolysaccharide (LPS)-induced Acute Lung Injury (ALI), NTP treatment attenuated tissue damage, inhibited the production of inflammatory cytokines, and promoted the anti-oxidative response. The supposed mechanism may be via suppressing the Toll-like receptor 4 (TLR4) signaling pathway. In conclusion, our study suggests a protective role of NTP in LPS-induced ALI by inhibiting inflammatory damage.