Novel and diverse mycoviruses co-infecting a single strain of the phytopathogenic fungus Alternaria dianthicola.

Alternaria dianthicola is a pathogenic fungus that causes serious leaf or flower blight on some medicinal plants worldwide. In this study, multiple dsRNA bands in the range of 1.2-10 kbp were found in a Alternaria dianthus strain HNSZ-1, and eleven full-length cDNA sequences of these dsRNA were obtained by high-throughput sequencing, RT-PCR detection and conventional Sanger sequencing. Homology search and phylogenetic analyses indicated that the strain HNSZ-1 was infected by at least nine mycoviruses. Among the nine, five viruses were confirmed to represent novel viruses in the families Hypoviridae, Totiviridae, Mymonaviridae and a provisional family Ambiguiviridae. Virus elimination and horizontal transmission indicated that the (-) ssRNA virus, AdNSRV1, might be associated with the slow growth and irregular colony phenotype of the host fungus. As far as we know, this is the first report for virome characterization of A. dianthus, which might provide important insights for screening of mycovirus for biological control and for studying of the interactions between viruses or viruses and their host.

A Panax notoginseng phosphate transporter, PnPht1;3, greatly contributes to phosphate and arsenate uptake.

The crisis of arsenic (As) accumulation in rhizomes threatens the quality and safety of Panax notoginseng (Burk.) F.H. Chen, which is a well-known traditional Chinese herb with a long clinical history. The uptake of arsenate (AsV) could be suppressed by supplying phosphate (Pi), in which Pi transporters play important roles in the uptake of Pi and AsV. Herein, the P . notoginseng Pi transporter-encoding gene PnPht1;3 was identified and characterised under Pi deficiency and AsV exposure. In this study, the open reading frame (ORF) of PnPht1;3 was cloned according to RNA-seq and encoded 545 amino acids. The relative expression levels revealed that PnPht1;3 was significantly upregulated under phosphate deficiency and AsV exposure. Heterologous expression in Saccharomyces cerevisiae MB192 demonstrated that PnPht1;3 performed optimally in complementing the yeast Pi-transport defect and accumulated more As in the cells. Combined with the subcellular localisation prediction, it was concluded that PnPht1;3 encodes a functional plasma membrane-localised transporter protein that mediates putative high-affinity Pi/H+ symport activity and enhances the uptake of Pi and AsV. Therefore, a better understanding of the roles of the P . notoginseng Pi transporter could provide new insight for solving As accumulation in medicinal plants.

Identification and First Report of Fusarium andiyazi Causing Sheath Rot of Zizania latifolia in China.

Zizania latifolia is a perennial plant native to East Asia. The swollen culm of Z. latifolia is a popular vegetable and traditional herbal medicine consumed in China and some other Asian countries. From 2019 to 2021, a sheath rot disease was found in Zhejiang Province of China. Symptoms mainly occurred in the leaf sheath showing as brown necrotic lesions surrounded by yellow halos. The pathogen fungal isolates were isolated from the affected sheaths. Ten representative isolates were selected for morphological and molecular identification by phylogenetic analyses of the translation elongation factor 1-α (TEF1) and the RNA polymerase II subunit beta (RPB2) gene regions. Based on the combined datasets, the fungal isolates were identified as Fusarium andiyazi. Koch's postulates were confirmed by pathogenicity test, re-isolation and re-identification of the fungal isolates. To the best of our knowledge, this is the first report of sheath rot caused by F. andiyazi in Z. latifolia in China.

Identification of species and materia medica within Saussurea subg. Amphilaena based on DNA barcodes.

Saussurea is one of the most species-rich genera in the family Asteraceae, where some have a complex evolutionary history, including radiation and convergent evolution, and the identification of these species is notoriously difficult. This genus contains many plants with medical uses, and thus an objective identification method is urgently needed. Saussurea subg. Amphilaena is one of the four subgenera of Saussurea and it is particularly rich in medical resources, where 15/39 species are used in medicine. To test the application of DNA barcodes in this subgenus, five candidates were sequenced and analyzed using 131 individuals representing 15 medical plants and four additional species from this subgenus. Our results suggested that internal transcribed spacer (ITS) + rbcL or ITS + rbcL + psbA-trnH could distinguish all of the species, while the ITS alone could identify all of the 15 medical plants. However, the species identification rates based on plastid barcodes were low, i.e., 0% to 36% when analyzed individually, and 63% when all four loci were combined. Thus, we recommend using ITS + rbcL as the DNA barcode for S. subg. Amphilaena or the ITS alone for medical plants. Possible taxonomic problems and substitutes for medicinal plant materials are also discussed.

P absorption and removal mechanism of new Salix clone (A42)on eutrophic water with different P concentrations.

Phosphorus is the necessary element for plant growth, and its concentration is one of the main indices for water eutrophication. Hence, it is significant to understand how woody plants purify phosphorus in eutrophic water. The purpose of this study is to reveal the P absorption and removal mechanism of Salix matsudana in eutrophic water with different P concentration. We selected new S. matsudana clone (A42) as experimental material and set three levels of P concentration (low P: 0.1, 0.2 mg·L-1; medium P: 1.0, 2.0 mg·L-1, high P: 10.0 mg·L-1), and the floating bed hydroponic experiment was conducted at the greenhouse from July to September, 2017. We found that S. matsudana efficiently removed P in water (removal rate >79% in 21 days). There was a positive correlation between the removal quantity and P concentration in the water. The removal ratio rose at first and then fell with increasing P. Owing to the purification of S. matsudana, the P concentrations ranging from 0.1 mg·L-1 to 1.0 mg·L-1 were reduced to minimum threshold concentration of eutrophication (0.016-0.032 mg·L-1) in seven days. The percentage of phosphorus input in water that assimilated by S. matsudana ranged from 29.0% to 66.9%. The quantity and ratio of assimilated P were respectively positive and negative relation with P concentration. Salix matsudana adapted to eutrophic water with different P concentrations and normally grew during experiment period, with root-shoot ratio being significantly increased with decreases of water P concentrations. The characteristic of phosphorus distribution in plant organs was: stem> leave>root, while the translocation factors (TF) of nitrogen and phosphorus were both greater than 3. When S. matsudana grew in eutrophic water with high phosphorus concentration, the TF of nitrogen and phosphorus significantly increased to 4.53±0.24 and 4.92±0.62 respectively. Our results indicated that S. matsudanais could purify the eutrophic water and it could normally grow. New clone of S. matsudana could effectively absorb phosphorus in the water and accumulated it in the stem, which could reduce secondary pollution. In conclusion, S. matsudana could be used for a short-term treatment on the eutrophic water with low P concentration, while for the long-term treatment it is adapted to eutrophic water with high phosphorus concentrations.

Differential responses of soil nematode community to pig manure application levels in Ferric Acrisols.

Excessive pig manure application probably degrades arable soil quality in some intensive pig farming areas. The responses of the nematode community to dosages of pig manure were investigated in Ferric Acrisols under 3-season peanut monoculture. Varying dosages of manure (1.75, 3.5, 7, 14 and 28 t·ha-1·yr-1) in combination with chemical fertilizer were applied to field plots, and chemical fertilizer alone was also applied as a control. With increasing manure application, the abundance of bacterivores and omnivores-predators increased, the abundance of plant parasites decreased, and fungivores abundance exhibited hump-shaped variation. Simpson diversity index and plant parasite index/maturity index of the nematode communities increased to a maximum level at a manure application rate of 3.5 t·ha-1·yr-1 and then sharply decreased. The changes in the soil nematode community were further determined to be correlated with chemical properties; available phosphorus had the strongest quadratic correlation with the two indices, implying that available phosphorus had a better indicative effect than other soil properties to nematode community. Available phosphorus in soil was deduced from 49 to 64 mg·kg-1 with the best nematode communities. Our results emphasized the importance of regular applications of manure in agriculture field to balance nematode diversity and build healthy agro-ecosystems.

Effect of brachytherapy on NF-κB and VEGF in gastric carcinoma xenografts.

Iodine-125 (125I) seed irradiation can be used as an important supplementary treatment for unresectable advanced gastric cancer. However, the radiobiological mechanism underlying brachytherapy remains unclear. Therefore, we investigated the influence of continuous and low-energy 125I irradiation on the cell cycle distribution, apoptosis, expression of NF-κB and VEGF and tumor growth in a human gastric cancer xenograft model. To create an animal model of gastric cancer, SGC-7901 cells were surgically implanted into mice. The 60 mice bearing SGC-7901 gastric cancer xenografts were randomly separated into 2 groups. Sham seeds (0 mCi) were implanted into the control group (n=30); 125I seeds (0.6 mCi) were implanted into the treatment group (n=30). At 28 days after irradiation, apoptosis was detected by flow cytometry. fluorescence micrograph detected intense VEGF and NF-κB immunofluorescence in the tumor samples, and changes in NF-κB and VEGF mRNA and protein expression were assessed by real-time PCR and western blot analysis, respectively. The tumor volume and weight were measured 0-28 days after 125I seed implantation. 125I seed irradiation induced significant apoptosis and G2/M phase arrest. Reduction in the intensities of VEGF and NF-κB immunofluorescence in tumor vessels was observed after treatment. NF-κB and VEGF mRNA and protein expression levels were substantially lower in the implantation treatment group than in the control group. Consequently, 125I seed implantation inhibited cancer growth and reduced cancer volume. The present study revealed that 125I seed irradiation significantly induced apoptosis and cell cycle arrest in the human gastric cancer xenografts. 125I-induced changes in NF-κB and VEGF expression are suggested as potential mechanisms underlying effective brachytherapy.

Clinical application of entire gastrointestinal barium meal combined with multi-temporal abdominal films in patients with intestinal neuronal dysplasia type B.

OBJECTIVE: To report and evaluate the application of entire gastrointestinal barium meal combined with multi-temporal abdominal films in the diagnosis of patients with intestinal neuronal dysplasia type B (IND type B). METHODS: Thirty-six patients with symptoms of long-standing constipation were enrolled in this study. The study took place at the Department of General Surgery, Xiangyang Central Hospital, Hubei Province, China from July 2007 to October 2012. All of them had already been subjected to the tests of barium enema and anorectal manometry and were suspected to be IND type B, but were not confirmed by mucous membrane acetylcholinesterase determination. All underwent the entire gastrointestinal barium meal combined with multi-temporal abdominal films. The data was collected and then analyzed retrospectively. RESULTS: After entire gastrointestinal barium meal combined with multi-temporal abdominal films, 30 out of 36 cases in this group were diagnosed with intestinal neuronal diseases, and then were treated with appropriate surgical treatment. The postoperative pathological diagnosis was IND type B. The other 6 patients in this group still could not be diagnosed explicitly after the test; thus, we treated them with conservative treatment. CONCLUSION: Entire gastrointestinal barium meal combined with multi-temporal abdominal films has the advantage of being able to test the gastrointestinal transfer capabilities and to find physiological and pathological changes simultaneously. It could provide important proof for the diagnosis of patients with intestinal neuronal dysplasia type B.

[Current studies and prospect of anti-thrombotic effect of Cordyceps fungi].

Cordyceps is an entomogenous fungi developed after being infected by insects or arthropods. Many varieties of entomogenous fungus show a huge application prospect in functional food and biological medicine fields. Among them, famous herb C. sinensis is their most excellent representative, and mainly used to replenish kidneys and moisten lungs, treating fatigue, night sweating, cardiovascular diseases, respiratory diseases, hypertension, hyperlipidemia, hyperglycemia, renal failure and so on. Currently, as a medicinal resource, C. sinensis has aroused attention from the world. In the past 20 years, many studies found that Cordyceps fungi show the anti-thrombotic effect, as well as a good potential for becoming a patent medicine. This article summarizes studies on Cordyceps fungi's effects in reducing blood viscosity and resisting platelet aggregation and anticoagulant, so as to provide thought for developing and utilizing these resources as anti-thrombotic medicines.

[Effects of transgenic insect-resistant cotton root exudates on the growth of Verticillium dahliae Kleb].

The decline of disease resistance of transgenic insect-resistant cotton has become one of the important factors restricting China cotton production. With two transgenic insect-resistant cotton varieties and their parental conventional cotton lines as test materials, the effects of their root exudates on the spore germination and mycelial growth of Verticillium dahliae Kleb were studied, and the components and contents of the amino acids and sugars in the exudates were determined. Comparing with their parental lines, the two transgenic insect-resistant varieties had a declined resistance against V. dahliae, and their root exudates had promotion effects on V. dahliae spore germination and mycelial growth. The root exudates of transgenic cotton Zhong-41, compared with those of its parent line Zhong-23, contained two additional amino acids Met and Lys, and the contents of Asp, Glu, Ser, Ala, Val, Leu, and Tyr were significantly higher. Though the components of amino acids in the root exudates of Simian-3 and transgenic cotton GK-12 were the same, the Tyr, Val, and Leu contents in the root exudates of transgenic cotton GK-12 were significantly lower. Four kinds of sugar were detected in the root exudates of Zhong-41, while only glucose was detected in the root exudates of Zhong-23. These four kinds of sugar were also detected in the root exudates of Simian-3 and GK12, but their contents differed significantly.

[Controlled clinical trials of therapeutic effects of Chinese herbs promoting blood circulation and removing blood stasis on the treatment of reflex sympathetic dystrophy with type of stagnation of vital energy and blood stasis].

OBJECTIVE: To observe clinical results of Chinese herbs promoting blood circulation and removing blood stasis on the treatment of reflex sympathetic dystrophy (RSD) with type of stagnation of vital energy and blood stasis. METHODS: RSD with type of stagnation of vital energy and blood stasis was distinguished as erubescence, high temperature, perspiration, damp and acro-edema, with middle level pain. From 2006 to 2008, 58 patients with RSD of stagnation of vital energy and blood stasis were randomly divided into the treatment group (30 cases) and the control group (28 cases). The former were treated with Chinese medicine to activate blood circulation and improve bone and muscle nourishment. Chinese medicine includes: Caesalpinia Sappan 10 g, Ligusticum Chuanxiong 6 g, Frankincense 6 g, Angelica 10 g, Safflower 6 g, Myrrh 6 g, Ground Beetle 10 g, Araliaceae 3 g, Radix Paeoniae Rubra 10 g, Pericarpium Citri Reticulatae 5 g, Lawn Pennywort Herb 15 g, Manis Pentadactyla 10 g, Corydalis Yanhusuo 10 g, Rhizoma Drynariae 15 g, which were boiled into decoction and the patients were take orally everyday with a course of treatment for 10 days, together with the boiled Chinese traditional medicine of stretching muscle and activating blood circulation to fume and wash the limbs twice everyday. The compatibility of medicines in prescription includes: Lycopodium Japanicum Grass 10 g, Gentiana Macrophylla Pall 10 g, Radix Angelicae Pubescentis 10 g, Angelica 10 g, Uncaria 10 g, Frankincense 6 g, Myrrh 6 g, Safflower 6 g. Control group were treated with a placebo of the same color for oral use and external application. The delivery times, method and the time of therapy were all the same as the treatment group. After 30 days' treatment, the effective indexes of VAS pain score and swelling condition were observed in both groups. RESULTS: VAS pain score: the treatment group decreased (3.8 +/- 0.8) points and the control group decreased (1.0 +/- 0.3) points, the difference between the two groups was significantly (P < 0.01). There was significantly difference in volume decrease of the swelling limb between treatment group (21.8 +/- 2.5) ml and the control group (10.3 +/- 2.1) ml (P < 0.01). The efficiency difference between treatment group and control group was significantly(P<0.01). CONCLUSION: With the different treatment based on different syndrome and emphasis on the nourishment of bone and soft tissue, treated by Chinese medicine to promote blood circulation and remove blood stasis in stagnation of vital energy and blood stasis, RSD get a favorable result.