Abundant and diverse endophytic bacteria associated with medicinal plant Arctium lappa L. and their potential for host plant growth promoting.

Arctium lappa L. is one of the medicinal and food homologous plants in China, which is rich in nutrients and medicinal ingredients. The use of plant growth-promoting (PGP) endophytic bacteria is an alternative to reducing chemical fertilizers in agricultural production. The aim of this study was to analyze the diversity of endophytic bacteria in different cultivars of A. lappa L. collected from two geographical locations in China and evaluate PGP traits of the isolates and their potential PGP ability in greenhouse condition. Endophytic bacterial community was investigated by culture-dependent and culture-independent methods. Isolates were screened and investigated for multiple PGP traits, and representative strains were inoculated host seedlings to evaluate the growth promoting effect. A total of 348 endophytic bacteria were obtained and they were distributed into 4 phyla and 30 genera. In addition, high throughput sequencing revealed more abundant bacterial community, including 17 bacterial phyla and 207 genera. A high proportion of PGP traits were detected, including production of indole acetic acid, siderophore, ammonia and phosphate solubilization. Four representative strains with multiple PGP traits of the most dominant genera (Bacillus, Pantoea, Microbacterium and Pseudomonas) were further selected for host inoculation and growth promoting evaluation, and they significantly increase seedlings length, root length and fresh weight. This study demonstrated that A. lappa L. harbors abundant endophytic bacteria, and some endophytic bacteria showed good potential for the development of microbial fertilizer in the future.

Targeted therapy of atherosclerosis by pH-sensitive hyaluronic acid nanoparticles co-delivering all-trans retinal and rapamycin.

Atherosclerosis, the leading cause of death in the elderly worldwide, is typically characterized by elevated reactive oxygen species (ROS) levels and a chronic inflammatory state at the arterial plaques. Herein, pH-sensitive nanoparticles (HRRAP NPs) co-delivering all-trans retinal (ATR), an antioxidant linked to hyaluronic acid (HA) through a pH-sensitive hydrazone bond, and rapamycin (RAP), an anti-atherosclerotic drug loaded into the nanoparticle core, are developed for targeted combination therapy of atherosclerosis. In this way, HRRAP NPs might simultaneously reduce ROS levels via ATR antioxidant activity and reduce inflammation via the anti-inflammatory effect of RAP. In response to mildly acidic conditions mimicking the lesional inflammation in vitro, HRRAP NPs dissociated and both ATR and RAP were effectively released. The developed HRRAP NPs effectively inhibited pro-inflammatory macrophage proliferation, and displayed dose- and time-dependent specific internalization by different cellular models of atherosclerosis. Also, HRRAP NP combination therapy showed an efficient synergetic anti-atherosclerotic effect in vitro by effectively inhibiting the inflammatory response and oxidative stress in inflammatory cells. More importantly, HR NPs specifically accumulated in the atherosclerotic plaques of apolipoprotein E-deficient (ApoE-/-) mice, by active interaction with HA receptors overexpressed by different cells of the plaque. The treatment with HRRAP NPs remarkably inhibited the progression of atherosclerosis in ApoE-/- mice which resulted in stable plaques with considerably smaller necrotic cores, lower matrix metalloproteinase-9, and decreased proliferation of macrophages and smooth muscle cells (SMCs). Furthermore, HRRAP NPs attenuated RAP adverse effects and exhibited a good safety profile after long-term treatment in mice. Consequently, the developed pH-sensitive HRRAP NP represent a promising nanoplatform for atherosclerosis combination therapy.

Antifungal Volatile Organic Compounds from Streptomyces setonii WY228 Control Black Spot Disease of Sweet Potato.

Volatile organic compounds (VOCs) produced by microorganisms are considered promising environmental-safety fumigants for controlling postharvest diseases. Ceratocystis fimbriata, the pathogen of black spot disease, seriously affects the quality and yield of sweet potato in the field and postharvest. This study tested the effects of VOCs produced by Streptomyces setonii WY228 on the control of C. fimbriata in vitro and in vivo. The VOCs exhibited strong antifungal activity and significantly inhibited the growth of C. fimbriata. During the 20-day storage, VOC fumigation significantly controlled the occurrence of the pathogen, increased the content of antioxidants and defense-related enzymes and flavonoids, and boosted the starch content so as to maintain the quality of the sweet potatoes. Headspace analysis showed that the volatiles 2-ethyl-5-methylpyrazine and dimethyl disulfide significantly inhibited the mycelial growth and spore germination of C. fimbriata in a dose-dependent manner. Fumigation with 100 µL/L 2-ethyl-5-methylpyrazine completely controlled the pathogen in vivo after 10 days of storage. Transcriptome analysis showed that volatiles mainly downregulated the ribosomal synthesis genes and activated the proteasome system of the pathogen in response to VOC stress, while the genes related to spore development, cell membrane synthesis, mitochondrial function, and hydrolase and toxin synthesis were also downregulated, indicating that WY228-produced VOCs have diverse modes of action for pathogen control. Our study demonstrates that fumigation of sweet potato tuberous roots with S. setonii WY228 or use of formulations based on the VOCs is a promising new strategy to control sweet potato and other food and fruit pathogens during storage and shipment. IMPORTANCE Black spot disease caused by Ceratocystis fimbriata has caused huge economic losses to worldwide sweet potato production. At present, the control of C. fimbriata mainly depends on toxic fungicides, and there is a lack of effective alternative strategies. The research on biological control of sweet potato black spot disease is also very limited. An efficient biocontrol technique against pathogens using microbial volatile organic compounds could be an alternative method to control this disease. Our study revealed the significant biological control effect of volatile organic compounds of Streptomyces setonii WY228 on black spot disease of postharvest sweet potato and the complex antifungal mechanism against C. fimbriata. Our data demonstrated that Streptomyces setonii WY228 and its volatile 2-ethyl-5-methylpyrazine could be a candidate strain and compound for the creation of fumigants and showed the important potential of biotechnology applications in the field of food and agriculture.

Biodegradation of phenol by a halotolerant versatile yeast Candida tropicalis SDP-1 in wastewater and soil under high salinity conditions.

In this study, a novel halotolerant phenol-degrading yeast strain, SDP-1, was isolated from a coastal soil in Jiangsu, China, and identified as Candida tropicalis by morphology and rRNA internal transcribed space region sequence analysis. Strain SDP-1 can efficiently remove phenol at wide ranges of pH (3.0-9.0), temperature (20-40 °C), and NaCl (0-5%, w/v), as well as the tolerance of Mn2+, Zn2+ and Cr3+ in aquatic phase. It also utilized multiple phenol derivatives and aromatic hydrocarbons as sole carbon source and energy for growth. Free cells of SDP-1 were able to degrade the maximum phenol concentration of 1800 mg/L within 56 h under the optimum culture conditions of 10% inoculum volume, pH 8.0, 35 °C and 200 rpm agitation speed. Meanwhile, SDP-1 was immobilized on sodium alginate, and the capability of efficiently phenol degradation of free cells and immobilized SDP-1 were evaluated. Shortened degradation time and long-term utilization and recycling for immobilized SDP-1 was achieved compared to free cells. The 1200 mg/L of phenol under 5% NaCl stress could be completely degraded within 40 h by immobilized cells. In actual industrial coking wastewater, immobilized cells were able to completely remove 383 mg/L phenol within 20 h, and the corresponding chemical oxygen demand (COD) value was decreased by 50.38%. Besides, in phenol-contained salinity soil (3% NaCl), 100% of phenol (500 and 1000 mg/kg) removal efficiency was achieved by immobilized SDP-1 within 12 and 26 days, respectively. Our study suggested that versatile yeast Candida tropicalis SDP-1 could be potentially used for enhanced treatment of phenol-contaminated wastewater and soil under hypersaline or no-salt environmental conditions.

Complete Genome Sequence of the Biocontrol Agent Pseudomonas chlororaphis subsp. aureofaciens SPS-41.

Pseudomonas chlororaphis subsp. aureofaciens SPS-41 is a plant growth-promoting rhizobacterium with biocontrol potential that was isolated from the rhizosphere of sweet potato in Xuzhou, Jiangsu Province, China. Our previous study demonstrated that volatile organic compounds (VOCs) produced by SPS-41 inhibited black spot disease fungi Ceratocystis fimbriata in postharvest sweet potatoes and a variety of other plant pathogens, and the VOCs also displayed strong nematocidal activity. In order to further explore the application potential of this strain, we here report the complete genome sequence of strain SPS-41. The genome consists of one chromosome (6,757,898 bp) with a G+C content 63.10%, which contains 5,951 coding genes, 67 transfer RNA genes, 16 ribosome RNA genes, and 85 other non-coding RNA genes. No plasmid was detected. The information of the genome will provide resources for studying the biocontrol mechanism of this strain.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Fermentation conditions optimization, purification, and antioxidant activity of exopolysaccharides obtained from the plant growth-promoting endophytic actinobacterium Glutamicibacter halophytocola KLBMP 5180.

In this study, an endophytic actinobacterium Glutamicibacter halophytocola KLBMP 5180, was investigated for the production and antioxidant activity of exopolysaccharides (EPSs). First, the suitable fermentation time, temperature, inoculation volume, pH value, and the carbon and nitrogen sources for EPSs production were obtained using the one variable at a time method (OVAT). Then, a central composition design was used for fermentation conditions optimization to obtain the maximum EPS yield. The optimal medium and condition were as follows: 100 mL broth in 250 mL Erlenmeyer flasks, including 3.65 g/L maltose, 9.88 g/L malt extract, 3.40 g/L yeast extract, 1.41 g/L MnCl2, pH 7.5, culture temperature 28 °C, and 200 rpm for 7 days, which increased the yield of EPSs to 2.89 g/L. Two purified EPSs, 5180EPS-1 (MW 58.9 kDa) and 5180EPS-2 (10.5 kDa), comprising rhamnose, galacturonic acid, glucose, glucuronic acid, xylose, and arabinose, were obtained for chemical analysis and antioxidant evaluation. The scavenging ability and reducing power of the superoxide anion and hydroxyl radicals demonstrated the moderate in vitro antioxidant activities of the two EPSs, thus indicating their potential to be a new source of natural antioxidants. However, further structure elucidation and functional studies need to be continued.

[A survey on cerebral infarction/transient ischemic attack inpatients compliance with secondary stroke prevention and follow-up 90 days].

OBJECTIVE: Poor compliance with evidence-based-medicine guidelines could significantly influence the effect of stroke prevention strategies. The objectives of this survey are to investigate the current status in Beijing of cerebral infarction/TIA (transient ischemic attack) inpatients' adherence to the evidence-based-medicine secondary prevention and behavior modifications both at the discharge and at 90-day follow up. METHODS: The survey included cerebral infarction/TIA patients successively admitted from 1, Oct 2006 to 1, May 2007, used ACEI/ARB, Statin, anti-platelet agents therapy, accepted carotid endarterctomy or angioplasty or stent, and modified behaviors to stop smoking and control weight. Data of patients during hospitalization were obtained from the medical documents in the inpatient department of 7 centers. The detailed information included the medicine therapy, lifestyle modifications, patients' social demographic background (age, sex, education and way of payment), major risk factors for stroke, and ischemic event subtypes (TIA or cerebral infarction). Above information of part of patients were collected through telephone interview at 90-day follow-up if the patients were not able to visit the clinic. RESULTS: 708 cerebral infarction/TIA inpatients who didn't have contraindications were included. Among them, the patents who haven't used anti-platelet agents, nor taken statin, nor accepted ACEI/ARB were about 0.4%, 41.8% and 63.6% respectively. Neither 27% of the patients have accepted the instructions on stop smoking. While about 588 patients suitable to drug therapy were followed up at 90 days, but the percentage of these patients who haven't compliance on anti-platelet agents, statin, ACEI/ARB was 26.9%, 52.6% and 59.4% respectively. Only 66.9% of the smoking patients have received the doctor instructions on smoking cessation. CONCLUSION: Stroke recurrence rate in China is higher than that of average figures in western countries. One of the key reasons of high recurrence of stroke in China is the poor adherence to stroke secondary prevention guidelines. Thus, investigate current status of secondary prevention among hospitals in capital of China and promote strategies to enhance the adherence to the guidelines and fill the gap between the clinical practice and evidence-based medicine in China, which is able to lower the stroke recurrence and save lives are urgently needed. Physician should take great care of the patient's compliances on the evidenced-based-medicine secondary prevents and behavior modifications, and take effective measures to improve the compliances.