Hemerocallis Citrina Research Articles

Lecithin extraction optimisation and synthesis in Hemerocallis citrina Baroni

Hemerocallis citrina Baroni (daylily) is an important flower plant, consumed as food and used in traditional Chinese medicine. The daylily flower bud is rich in several functional substances, and lecithin is the most important among them. However, the optimum conditions for extracting lecithin from daylily flower buds are yet to be determined. The objective of this study was to examine the differences in the lecithin contents in daylily, identify the key enzymes and genes involved in lecithin synthesis, and establish the optimal conditions for lecithin extraction and detection by high-performance liquid chromatography. Initial treatment with acetone before ethanol extraction significantly increased the quantity of lecithin extracted compared with that extracted by direct ethanol extraction. The optimum lecithin extraction conditions were as follows: 90% ethanol concentration, 1:20 material-to-solvent ratio, 30 min extraction time, and 40°C extraction temperature. The highest lecithin content was obtained from the flower bud, specifically the stamen (3.49 mg/g). Additionally, the best harvest period for daylily flower buds with regards to lecithin content and nutritional quality was between 18 h before flowering and bloom (0 h). The lecithin content in freeze-dried daylily flower buds was higher than that in oven-dried and sun-dried flower buds. Among the varieties examined, Xiushan Huanghua (H0177) had the highest lecithin content (3.99 mg/g). Furthermore, the activity of phosphatidylethanolamine N-methyltransferase (PEAMT) was significantly higher than that of other enzymes involved in lecithin synthesis. Additionally, genes involved in lecithin synthesis were significantly expressed in the flower buds at different developmental stages. Overall, the findings of this study contribute to the understanding of lecithin synthesis in daylily.

Study and Experimental Validation of the Functional Components and Mechanisms of Hemerocallis citrina Baroni in the Treatment of Lactation Deficiency.

The function of Hemerocallis citrina Baroni (daylily) on promoting lactation is reported in several ancient Chinese medicine books. However, nowadays, there is no conclusive data to support this statement. In this study, we investigated the effect of Hemerocallis citrina Baroni extract (HCE) on lactation insufficiency in chronic unpredictable mild stress (CUMS) dams and further explored the mechanism and functional components through network pharmacology. The results showed that HCE could increase the offspring’s weight, serum prolactin (PRL), and oxytocin (OT) level of CUMS dams. Network pharmacology analysis revealed that the facilitation of HCE on lactation is the result of the comprehensive action of 62 components on 209 targets and 260 pathways, among this network, quercetin, kaempferol, thymidine, etc., were the vital material basis, signal transducer and activator of transcription 3 (STAT3), mitogen activity protein kinase 1 (MAPK1), tumor protein P53 (TP53), etc., were the core targets, and the prolactin signaling pathway was the core pathway. In addition, verification test results showed that HCE regulated the abnormal expression of the prolactin signaling pathway, including STAT3, cyclin D1 (CCND1), MAPK1, MAPK8, nuclear factor NF-kappa-B p105 subunit (NFKB1), and tyrosine-protein kinase (JAK2). In conclusion, HCE exhibited a facilitation of lactation insufficiency, in which quercetin, kaempferol, thymidine, etc., were the most important material basis. The mechanism of this promotional effect is mediated by the prolactin signaling pathway in mammary gland.

Dynamic changes of pectin epitopes and daylily tepals during flower opening and senescence of Hemerocallis citrina

Ephemeral flower deaths occur within 24 h of flowering in the genus Hemerocallis. The senescence of plant organs is owing to the loss of cell wall cohesion. Pectin is the most structurally complex plant cell wall polysaccharide and has an important role in cell-to-cell adhesion. In this study, we used histological stains, immunolocalization and pectin characterization analyses to examine the dynamic changes in pectin in Hemerocallis citrina ‘Datonghuanghua’ tepals at eight stages (S1–S8). The pectin-specific antibodies JIM5 and JIM7, which hybridize to differently esterified regions of the homogalacturonan (HG) backbone, showed that demethylesterification of HG-type pectins was evident after the outer tepals had opened. Before flower opening, these two antibodies labelled tissue continuously across the cell wall, and the cells were round and tightly packed. The labeling of highly esterified epitopes recognized by JIM7 became deformed when the outer tepals opened. However, 12 h after blooming, low-and high-esterified epitope signals were disrupted near the vascular cylinder, and air space then appeared until these signals disappeared. labeling with the antibody LM5, which is specific for the (1 → 4)-β galactan side chains of rhamnogalacturonan-I, was disrupted near the vascular cylinder at the same stage. Moreover, the contents of neutral sugars in pectin analyses showed that galactose decreased significantly at 12 h after blooming. Three enzymes were involved in pectin remodeling at different developmental phases. The activities of pectin methylesterase continued to increase after the outer tepals opened and reached a significant peak at 12 h after blooming. The activities of polygalacturonase and pectate lyase also increased, but the difference was not significant. Our findings revealed that demethylesterification of HG-type pectins was evident after the outer tepals opened, and this remodeling greatly facilitated their degradation by pectic enzymes. The loss of cell wall material and cohesion that accompanied the activities of pectic enzymes was associated with senescence. Therefore, remodeling of the pectin in cell walls of the tepals is necessary for the occurrence of senescence.

Optimization of ultrasonic-assisted extraction of polysaccharides from Hemerocallis citrina and the antioxidant activity study.

The present study discussed the optimization of the ultrasonic-assisted extraction of polysaccharides from daylily polysaccharides (DPs). The extracted crude polysaccharides were further separated and purified, and the antioxidant activities including 1,1-diphenyl-2-111 picrylhydrazyl (DPPH) radical scavenging, 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radical scavenging, hydroxyl radical scavenging, and ferric-reducing antioxidant power (FRAP) activities of the obtained fractions were also evaluated. The results showed that the optimal ultrasonic-assisted extraction parameters with DPs yield of 15.25 ± 1.13% were water to powder ratio of 25 ml/g, extraction power of 694 W, extraction temperature of 71°C, extraction time of 38 min, and three times extraction. By DEAE Sepharose Fast Flow column, four water-soluble polysaccharide fractions (DP-1, DP-2, DP-3, and DP-4) were successfully obtained. Monosaccharide component analysis showed that the four obtained fractions were all hetero-polysaccharides that mainly contained rhamnose, arabinose, fructose, galactose, glucose, galacturonic acid, and glucuronic acid in different molar ratios. All the four DP fractions did show obvious antioxidant activities in vitro, and the DP-3 component had relatively high ABTS free radical scavenging activity. Overall, our research showed that DPs could provide cheap raw materials for the development of natural antioxidants in medicines, functional foods, and even cosmetics. PRACTICAL APPLICATION: This article deals with the optimization of the ultrasonic-assisted extraction of polysaccharides from daylily and its antioxidant activities. The results showed that the optimal ultrasonic-assisted extraction yield of DPs was 15.25 ± 1.13%. By DEAE Sepharose Fast Flow column, four water-soluble polysaccharide fractions were successfully obtained, and all the four DP fractions did show obvious antioxidant activities in vitro. Daylily polysaccharides could provide cheap raw materials for the development of natural antioxidants in medicines, functional foods, and even cosmetics.

Particularitățile morfobiologice ale unor specii din genul Hemerocallis L.

In the National Botanical Garden (Institute) “Al. Ciubotaru”, four species from the family Xanthorrhoeaceae, subfamily Hemerocallidoideae were studied: Hemerocallis citrina Baroni, H. lilioasphodelus L., H. minor Mill, H. Middendofii Trautv.&C.A. Mey. The characteristics of growth, development and ontogenesis were established.

Study on the Sleep-Improvement Effects of Hemerocallis citrina Baroni in Drosophila melanogaster and Targeted Screening to Identify Its Active Components and Mechanism.

Hemerocallis citrina Baroni (HC) is an edible plant in Asia, and it has been traditionally used for sleep-improvement. However, the bioactive components and mechanism of HC in sleep-improvement are still unclear. In this study, the sleep-improvement effect of HC hydroalcoholic extract was investigated based on a caffeine-induced insomnia model in Drosophila melanogaster (D. melanogaster), and the ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-ESI-Orbitrap-MS) and network pharmacology strategy were further combined to screen systematically the active constituents and mechanism of HC in sleep-improvement. The results suggested HC effectively regulated the number of nighttime activities and total sleep time of D. melanogaster in a dose-dependent manner and positively regulated the sleep bouts and sleep duration of D. melanogaster. The target screening suggested that quercetin, luteolin, kaempferol, caffeic acid, and nicotinic acid were the main bioactive components of HC in sleep-improvements. Moreover, the core targets (Akt1, Cat, Ple, and Sod) affected by HC were verified by the expression of the mRNA of D. melanogaster. In summary, this study showed that HC could effectively regulate the sleep of D. melanogaster and further clarifies the multi-component and multi-target features of HC in sleep-improvement, which provides a new insight for the research and utilization of HC.

The chromosome-level Hemerocallis citrina Borani genome provides new insights into the rutin biosynthesis and the lack of colchicine

Hemerocallis citrina Borani (huang hua cai in Chinese) is an important horticultural crop whose flower buds are widely consumed as a delicious vegetable in Asia. Here we assembled a high-quality reference genome of H. citrina using single-molecule sequencing and Hi-C technologies. The genome assembly was 3.77 Gb and consisted of 3183 contigs with a contig N50 of 2.09 Mb, which were further clustered into 11 pseudochromosomes. A larger portion (3.25 Gb or 86.20%) was annotated as a repetitive content and 54,295 protein-coding genes were annotated in the genome. Genome evolution analysis showed that H. citrina experienced a recent whole-genome duplication (WGD) event at ~15.73 million years ago (Mya), which was the main factor leading to many multiple copies of orthologous genes. We used this reference genome to predict 20 genes involved in the rutin biosynthesis pathway. Moreover, our metabolomics data revealed neither colchicine nor its precursors in H. citrina, challenging the long-standing belief that this alkaloid causes poisoning by the plant. The results of our disruptive research are further substantiated by our genomic finding that H. citrina does not contain any genes involved in colchicine biosynthesis. The high-quality genome lays a solid foundation for genetic research and molecular breeding of H. citrina.

First report of Epicoccum sorghinum causing leaf spot on Hemerocallis citrina in China.

Hemerocallis citrina Baroni, also called yellow flower vegetable (huang hua cai in Chinese), is belonging to the family Xanthorrhoeaceae and is widely planted in China, the Korea Peninsula and Japan for ornamental purposes and vegetable value. In addition, they could also be used as a traditional Chinese medicinal and modern medicinal plant (Du et al. 2014). In August 2019, a leaf spot disease was observed on H. citrina plants in Zhejiang Province of China, with approximately 85% incidence in almost 700 ha. Symptoms were firstly displayed as small, water-soaked, pale chlorotic spots, with yellow halos enlarged into large fusiform spots with brown edge and gray centers. Later, infected leaves were badly damaged and became wilted. Small pieces of infected tissue were excised from the margin of necrotic lesions, surface disinfected with 70% ethanol for 8s, 0.1% HgCl2 for 1 min, rinsed with sterile distilled water for three times, and incubated on potato dextrose agar (PDA, amended with 100 mg/L streptomycin sulfate) at 26°C in the dark. Fungal colonies with similar cultural morphology were consistently obtained from repeated isolations. When cultured on PDA, colonies were villose, regular, grayish-green, and turned gray-brown, with the reverse side became reddish-brown. Chlamydospores were gray, unicellular or multicellular, nearly spherical, 11 to 27 × 10 to 23 μm. Pycnidia and conidia were produced on PDA when the fungal colonies were exposed to ultraviolet light for 12 h with a distance of 40 cm to the late source. Pycnidia were brown, mostly spheroid, and measured 90 to 138 × 120 to 210 μm. Conidia were hyaline, ellipsoidal, unicellular, aseptate, 4.3 to 5.5 × 1.8 to 2.4 μm. These morphological characteristics agreed with the descriptions of Epicoccum sorghinum (Zhou et al. 2018). The DNA of a representative strain HHC6-2 was extracted using CTAB method and the rDNA internal transcribed spacer (ITS), actin (ACT) and β-tubulin (TUB) genes were amplified and sequenced, using the primers ITS4/ITS5 (White et al. 1990), ACT512F/ACT783R (Carbone and Kohn 1999) and Bt-1/Bt-2 (Glass and Donaldson 1995), respectively. BLASTn searches of the resulting ITS, ACT and TUB sequences (accession nos. MW073403, MW080522, MW080521) revealed 98.58 to 100% identity to the E. sorghinum sequences (MT125854, MN956831 and MF987525). The pathogenicity test was carried out by inoculation of potted H. citrina plants using conidial suspensions. H. citrina seedlings were planted in pots with sterilized soil. Before inoculation, leaves were surface-disinfected with 70% ethanol and sterile distilled water. Leaves were inoculated by placing small droplets of conidial suspensions (105 conidia/ml) on one side of the midvein, and 3 to 5 drops were used per leaf. Sterile water was used as control. All the inoculated plants were placed in humid chambers at 25°C for 48h, and then maintained in a greenhouse at 25°C with a 16 h day-8 h night cycle. The pathogenicity assays were performed twice with three replications. Four days after inoculation, yellow to brown spots resembling those observed in the fields developed on the inoculated leaves. However, no symptoms were observed on the controls. E. sorghinum was re-isolated and identified based on morphological and molecular techniques as described above. To our knowledge, this is the first report of E. sorghinum causing leaf spot on H. citrina. It seems to be a threat for H. citrina planting in China and should be considered in order to reduce losses caused by this disease. This study might provide the basis for diagnosis and control of the disease.

Intra-leaf heterogeneities of hydrogen isotope compositions in leaf water and leaf wax of monocots and dicots

Several recent studies showed that leaf wax n-alkane δ2H values (δ2Hwax) within a leaf were heterogeneous in a small number of species. It still remains unclear whether the heterogeneity of intra-leaf δ2Hwax values is general for various species, how δ2Hwax values vary spatially and temporally, and whether there is a common explanation for the intra-leaf δ2Hwax heterogeneity in higher plants. Here we compared the hydrogen isotope compositions of leaf wax and corresponding leaf water (δ2Hlw) across leaf sections among a variety of monocot and dicot plant species. There is significant and consistent heterogeneity for both δ2Hwax and δ2Hlw, i.e., base-to-tip 2H-enrichment for monocots (except Hemerocallis citrina, and Dactylis glomerata) whereas base-to-tip and center-to-edge increases in δ2Hwax and δ2Hlw for dicots. The consistent occurrence of variations of δ2Hlw and δ2Hwax values within a leaf imply that δ2Hwax values probably inherit point-to-pint from in-situ δ2Hlw values, and thus the intra-leaf δ2Hwax heterogeneity mainly results from the spatial pattern of intra-leaf δ2Hlw values associated with veinal structures between dicots and monocots. The general heterogeneity of intra-leaf δ2Hwax values further intensifies that it is necessarily needed for in-depth understanding leaf wax biomarker.

Quantified Benefits of Cultivating Day-Lily (Hemerocallis citrina) Hedgerows over Multiple Years on Sloped Red-Soil Farmland, Southern China

Planting hedgerows is an effective measure for increasing soil and water conservation on sloped farmland, although the comprehensive and dynamic changes in benefits across the life span of the hedgerow are not fully understood. To expand our understanding in this area, day-lily (Hemerocallis citrina) was selected to research the effect of hedgerows built on sloped red-soil farmland in southern China. In situ observations of runoff and soil loss from 2012 to 2018 were recorded in triplicate for two treatments: (1) conventional downslope-ridge tillage (CT) and (2) hedgerow intercropping with downslope-ridge tillage (HT). Micro-topography, soil nutrient content, and crop yield changes in the study plots were recorded. Hedgerows began intercepting runoff and soil loss simultaneously from the second year after they were planted, reducing annual surface runoff (35–66%) and annual erosive sediment (32–87%). As hedgerows became established, they provided increasing positive effects on soil and water conservation. However, those beneficial effects were gradually obscured by changes in micro-topography. The presence of the hedgerow had no obvious influence on soil total nutrient contents under this experimental condition. On average, planting the hedgerow reduced peanut yield by 21% but reduced rapeseed yield only by 2%. Compared with CT treatment, the average economic output of HT treatment after reaching the hedge’s initial production was 1.52 times greater, showing a quadratic trend and reaching a maximum about 4 years after construction by regression. These findings indicate that hedgerow management could be optimized according to the desired beneficial soil and water conservation or crop yield characteristics in different developmental phases.

Systematic identification metabolites of Hemerocallis citrina Borani by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with a screening method

Hemerocallis citrina Borani is an important crop and its flower buds are widely consumed in East Asian areas as a vegetable, as well as in traditional Chinese medicine, due to its health-promoting properties. Metabolites present in plant-derived foods or medicines are in part responsible for their desirable flavor profiles and health benefits. Nevertheless, detailed information about these compounds in H. citrina is scarce. Therefore, this study aimed to investigate the metabolites by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). In this study, a total of 144 compounds, including 14 amides, 25 polyphenols, 44 flavonols, 35 anthraquinones, 15 naphthols, and 11 other components, were detected by the established screening method and were identified by their precise m/z values, characteristic tandem mass spectrometry (MS/MS) data and fragmentation pathways of references, 111 of which were reported in this plant for the first time. The distribution of identified ingredients in different parts of H. citrina was determined. Interestingly, colchicine, which had been reported as a toxic compound in the fresh flower buds in previous studies and various news reports, was not found. This work marks the first comprehensive study of metabolites from commercial flower buds and different parts of H. citrina.

Urinary metabolomics analysis of the anti-depressive effects of Hemerocallis citrina extracts in a simulated microgravity-induced rat model of depression

Urinary metabolomics analysis of the anti-depressive effects of Hemerocallis citrina extracts in a simulated microgravity-induced rat model of depression

The complete chloroplast genome of Hemerocallis citrina (Asphodelaceae), an ornamental and medicinal plant

Hemerocallis citrina (Asphodelaceae) has been wildly cultivated as ornamental and medicinal plant. Here, we reported the first chloroplast genome sequence of H. citrina. The chloroplast genome size is 156,088 bp with GC content of 37.3%, including a large single-copy (LSC) of 84,843 bp, a small single-copy (SSC) of 18,507 bp, and a pair of 26,369 bp IR(inverted repeat) regions. A total of 133 genes were annotated including 87 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. The phylogenetic analysis revealed that H. citrina belongs to the Hemerocallis genus in Asphodelaceae family.

Transcriptome sequencing and expression profiling of genes involved in daylily (Hemerocallis citrina Borani) flower development

Daylily (Hemerocallis citrina Borani), a widespread ornamental plant around the world, has been considered as an ideal model to study flower development recently. To initiate the research to comprehensively investigate the molecular mechanism of the flower development of daylily, we performed a transcriptome sequencing study to establish the transcriptional profiles of this plant at different flower developmental stages. Briefly, we divided the flower development of daylily into six stages, and performed transcriptome sequencing for the samples collected from each stage. A total of 56,598 unique transcripts were obtained, of which 35,424 ones (about 62.6%) encode proteins with known functions based on homology searches against well-studied model plants. The expression profiles of these genes showed that 7,395 genes were differentially expressed during flower development. Further functional enrichment analysis for these differentially expressed genes revealed that they are enriched in the processes of translation, oxidation-reduction, and cell wall metabolism, suggesting the critical roles of these processes in daylily flower development.

AM真菌对采煤沉陷区黄花菜生长及根际土壤养分的影响

PDF HTML阅读 XML下载 导出引用 引用提醒 AM真菌对采煤沉陷区黄花菜生长及根际土壤养分的影响 DOI: 10.5846/stxb201701130106 作者: 作者单位: 中国矿业大学北京煤炭资源与安全开采国家重点实验室,河南省地质环境勘查院,陕西煤化集团柠条塔煤矿;陕西煤化集团柠条塔煤矿,中国矿业大学北京地球科学与测绘工程学院,中国矿业大学北京地球科学与测绘工程学院,中国矿业大学北京地球科学与测绘工程学院 作者简介: 通讯作者: 中图分类号: 基金项目: 国家自然科学基金项目（51574253）；首都科技领军人才工程（Z18110006318021） Effects of arbuscular mycorrhizal fungi on daylily growth and soil fertility in a coal mining subsidence area of northern Shaanxi Author: Affiliation: College of Geoscience and Surveying Engineering,China University of Mining and Technology Beijing,Henan Institute of Geo-Environment Exploration,Ningtiao Tower Mine of Shaanxi Coal Group,State Key Laboratory of Coal Resources and Safe Mining,China University of Mining and Technology Beijing,State Key Laboratory of Coal Resources and Safe Mining,China University of Mining and Technology Beijing,State Key Laboratory of Coal Resources and Safe Mining,China University of Mining and Technology Beijing Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:于陕北黄土沟壑采煤沉陷区内布设试验小区，对黄花菜（Hemerocallis citrina Baroni）接种丛枝菌根真菌（arbuscular mycorrhizal fungi，AMF）-摩西管柄囊霉菌，通过测定黄花菜光合性能、植株生长、抗逆性、土壤养分含量、根际微生物数量等，揭示AM真菌对黄花菜生长和土壤养分的影响。结果表明，黄花菜种植3-5个月后，接种AM真菌显著提高了黄花菜株高、冠幅及其根系菌根侵染率、菌丝密度。与不接种对照区相比，接种AM真菌后黄花菜叶片的光合速率、可溶性糖含量和过氧化氢酶活性分别提高了51%、12%、79%。接种AM真菌处理区黄花菜根际土壤的电导率、有机质、碱解氮和速效钾含量等均显著高于对照区，细菌数量和磷酸酶活性的菌根贡献率分别达77%和24%。表明采煤沉陷区扰动土壤接种AM真菌具有增强土壤微生物活性、改善土壤肥力和提高黄花菜植株抗逆性的作用，对促进陕北黄土沟壑采煤沉陷区经济作物生长和提高土壤质量具有重要现实生态意义。 Abstract:Effects of arbuscular mycorrhizal fungi (AM) on the growth of daylily (Hemerocallis citrina Baroni) and soil fertility were investigated by measuring daylily growth, soil nutrition contents, soil microbial population, and enzyme activities in a coal mining subsidence loess hilly area of northern Shaanxi. Results showed that AM fungal inoculation significantly increased plant height and crown width, mycorrhizal colonization rate, and hyphal density. Compared with control, AM fungal inoculation increased leaf photosynthesis rate, soluble sugar content, and catalase activity by 51%, 12%, and 79%, respectively. Moreover, the soil conductivity and organic matter, available nitrogen, and potassium content in the rhizosphere soil were significantly higher in mycorrhizal plants than control, and the contribution rates of AM fungal inoculation to bacteria population and acid phosphatase activity in rhizospheric soil were 77% and 24%, respectively. This showed that AM fungal inoculation may play an important role in enhancing plant resistance stress and improving soil fertility, soil microbial population, and enzyme activities. It will have an important value in promoting the growth of cash crops and further research on degraded land reclamation. 参考文献 相似文献 引证文献

First Report of Daylily Flower Rot Caused by Fusarium proliferatum in China

Daylily (Hemerocallis citrina Baroni) is an important plant whose flowers are commonly used as a vegetable and in medicine in China. From June 2015 to July 2016, brown and rotted flowers, some of which were covered with white mycelia, were observed in a 0.8-acre commercial daylily field in Yanqing District, Beijing, China. Disease incidence was ∼40%. A total of 24 diseased flowers were collected, cut into 0.5 cm diameter sections from the edge of the rot/healthy tissues, surface-sterilized with 1% NaClO for 1 min followed by 75% ethanol for 30 s, and then rinsed twice with sterilized distilled water. Five surface-sterilized sections per flower were placed on potato sucrose agar (PSA) and carnation leaf agar (CLA) (Leslie and Summerell 2006) and incubated for 5 days at 25°C with a 12-h light-dark cycle in a chamber. Seventy-eight singe-spore isolates with similar morphological features as determined by a microscope were obtained (Song et al. 2017), and 24 were used as the research materials and incubated on PSA and CLA. The colonies on PSA were initially white, becoming purple, with rose to dark purple pigment on the surface. The macroconidia were abundant on CLA, hyaline, straight to slightly curved, 3 to 5 septate, 21.4 to 49.3 × 2.3 to 4.9 µm. Microconidia were oval, obovoid, or ellipsoidal, 0 to 1 septate, 4.5 to 15.1 × 2.2 to 4.3 µm. The cultural and morphological characteristics of the fungus were similar with previous reports of Fusarium proliferatum (Ritieni et al. 1995). Genomic DNA of all 24 isolates were extracted using the plant genomic DNA kit (Tiangen, China). The small subunit ribosomal RNA (mtSSU), elongation factor 1 alpha (EF-1α), and beta-tubulin (Tub) regions were amplified using the primers NSM1/NSM2 (Li et al. 1994), EF-1Ha/EF-2Tb (Alastruey-Izquierdo et al. 2008), and Beta1/Beta2 (Geiser et al. 2004). The amplicon sizes were ∼534, 691, and 745 bp, respectively. Three sequences of isolate HYC1410080201 were submitted to GenBank (accessions MF442355, MF448528, and MF442354, respectively). Isolates had 100% similarity with sequences of several isolates of F. proliferatum in GenBank (KX218406, KY081546, and KR861512, respectively). Pathogenicity was tested and Koch’s postulates satisfied in a growth chamber by inoculating daylily flowers. Briefly, spores were collected from 7-day-old PDA cultures of each isolate (n = 24) maintained at 25°C and suspended in sterilized water at 1 × 10⁶ conidia/ml. Spore suspension, 5 ml from each isolate, was sprayed onto the flowers of a plant with 10 plants inoculated per fungal isolate. Control plants were sprayed with 5 ml of sterile water. Each isolate was replicated three times. Five days after inoculation, small, pale yellow, water-soaked spots were observed on flowers of all inoculated plants. Under high humidity conditions, inoculated flowers were covered with a layer of white mycelia. Infected flowers wilted and dropped off, similar to symptoms observed on plants in the field. No symptoms were observed on control plants. Isolates obtained from inoculated plants had identical morphological features to the original isolates. Thus, Koch’s postulates were fulfilled and showed that F. proliferatum was the causal agent. To our knowledge, this is the first report of F. proliferatum causing daylily flower rot worldwide. Since this disease occurs during flowering and seriously affects edible and medicinal value, its presence has economic implications for daylily production.

Evaluation of the toxicological properties and anti-inflammatory mechanism of Hemerocallis citrina in LPS-induced depressive-like mice.

Hemerocallis citrina Baroni (Liliaceae), a Liliaceae plant, has been widely used in food and traditional medicine. This study investigated the safety of ethanol extracts from Hemerocallis citrina (HCE) after oral treatment (p.o.) and evaluating the anti-inflammatory mechanism of HCE in a lipopolysaccharide (LPS)-induced depressive-like model. First, in an 8-week experimental procedure, blood and tissue samples collected from mice were used for biochemical and histopathological analysis every two weeks. Neither the body weight nor relative organ weights were affected by HCE administration. Only the total cholesterol levels were decreased by HCE administration. Histopathological analysis showed no significant liver and kidney changes caused by HCE. In addition, in an LPS-induced mouse depressive-like model, HCE significantly reversed the reduction of sucrose preference with LPS. The results also indicated that LPS activated the nuclear factor-κB (NF-κB), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in the prefrontal cortex. In contrast, these activations were normalized by HCE pretreatment. In summary, our study provided essential evidence for the safety of Hemerocallis citrina in both food and medicine. The results also demonstrated that HCE exhibited antidepressant-like effects that might be related to inhibition of the NF-κB signaling pathway.

Identification and validation of reference genes for quantitative real-time PCR studies in long yellow daylily, Hemerocallis citrina Borani.

Gene expression analysis using reverse transcription quantitative real-time PCR (RT-qPCR) requires the use of reference gene(s) in the target species. The long yellow daylily, Hemerocallis citrina Baroni. is rich in beneficial secondary metabolites and is considered as a functional vegetable. It is widely cultivated and consumed in East Asian countries. However, reference genes for use in RT-qPCR in H. citrina are not available. In the present study, six potential reference genes, actin (ACT), AP-4 complex subunit (AP4), tubulin (TUB), ubiquitin (UBQ), 18S and 60S ribosomal RNA, were selected and their expression stability in different developmental stages, organs and accessions was evaluated using four statistical software packages (geNorm, NormFinder, BestKeeper, and RefFinder). For commercial flower buds of different landraces, the combination of 60S, TUB, and AP4 was appropriate whereas ACT and 60S was suitable for normalization of different organs. In addition, AP4 exhibited the most stable expression in flower buds among different developmental stages. UBQ was less stable than the other reference genes under the experimental conditions except under different organs was 18S. The relative expression levels of two genes, primary-amine oxidase (HcAOC3) and tyrosine aminotransferase (HcTAT) which play important roles in alkaloid biosynthesis were also examined in different organs of the ‘Datong’ landrace, which further confirmed the results of selected reference genes. This is the first report to evaluate the stability of reference genes in the long yellow daylily that can serve as a foundation for RT-qPCR analysis of gene expression in this species.

Effects of phenolic constituents of daylily flowers on corticosterone- and glutamate-treated PC12 cells

BackgroundDaylily flowers, the flower and bud parts of Hemerocallis citrina or H. fulva, are well known as Wang-You-Cao in Chinese, meaning forget-one’s sadness plant. However, the major types of active constituents responsible for the neurological effects remain unclear. This study was to examine the protective effects of hydroalcoholic extract and fractions and to identify the active fractions.MethodsThe extract of daylily flowers was separated with AB-8 resin into different fractions containing non-phenolic compounds, phenolic acid derivatives and flavonoids as determined using UPLC-DAD chromatograms. The neuroprotective activity was measured by evaluating the cell viability and lactate dehydrogenase release using PC12 cell damage models induced by corticosterone and glutamate. The neurological mechanisms were explored by determining their effect on the levels of dopamine (DA), 5-hydroxy tryptamine (5-HT), γ-aminobutyric acid (GABA), noradrenaline (NE) and acetylcholine (ACh) in the cell culture medium measured using an LC-MS/MS method.ResultsPretreatment of PC12 cells with the extract and phenolic fractions of daylily flowers at concentrations ranging from 0.63 to 5 mg raw material/mL significantly reversed corticosterone- and glutamate-induced neurotoxicity in a dose-dependent manner. The fractions containing phenolic acid derivatives (0.59% w/w in the flowers) and/or flavonoids (0.60% w/w) exerted similar dose-dependent neuroprotective effect whereas the fractions with non-phenolic compounds exhibited no activity. The presence of phenolic acid derivatives in the corticosterone- and glutamate-treated PC12 cells elevated the DA level in the cell culture medium whereas flavonoids resulted in increased ACH and 5-HT levels.ConclusionPhenolic acid derivatives and flavonoids were likely the active constituents of daylily flowers and they conferred a similar extent of neuroprotection, but affected the release of neurotransmitters in a different manner.

Antidepressant-like effects and cognitive enhancement of the total phenols extract of Hemerocallis citrina Baroni in chronic unpredictable mild stress rats and its related mechanism

Ethnopharmacological relevanceDepression induce distressed emotional state and cognitive deficits simultaneously, which both should be improved in the treatment. Hemerocallis citrina Baroni (HC) is a traditional herbal medicine in Eastern-Asia areas and the total phenols extract of HC (HCPE) contains the main active ingredients. It has been reported that HC has the emotional improvement effect. But the cognitive effect of HC was seldom researched. Aim of the studyWe designed to evaluate the antidepressant and cognitive improvement effect of HCPE using a chronic unpredictable mild stress (CUMS) model, and the potential mechanisms were explored by investigating the corticosterone (CORT), monoamine neurotansmitters, brain-derived neurotropic factor (BDNF) and oxidative stress. Materials and methodsThe depression rats were induced by CUMS procedures and treated with HCPE (10, 20, 40mg/kg/day, by gastric gavage). The antidepressant effect was evaluated by sucrose preference test, open field test and body weight, while the cognitive improvement was investigated using morris water maze test. Besides, the levels of monoamine neurotransmitters in the hippocampus and frontal cortex were measured by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The serum CORT and BDNF in hippocampus were test using enzyme-linked immunosorbent assay (ELISA) kits. The oxidative stress indicators in frontal cortex were also analyzed. ResultsHCPE (40mg/kg) improved the emotion and cognition related behaviors in depression effectively. Moreover, HCPE increased the neurotransmitters concentration (5-HT, DA and NE) in the hippocampus and frontal cortex compared with CUMS rats. Meanwhile, the CUMS induced changes of serum corticosterone level and the hippocampus BDNF level were reversed. Besides, HCPE reduced malondialdehyde (MDA) in the frontal cortex of model rats. ConclusionIt suggested that HCPE could improve the depression-like emotional status and associated cognitive deficits in CUMS rats, which might be mediated by regulation of neurotransmitters and BDNF levels in brain, alleviation of corticosterone level as well as the alleviation of oxidative stress.