TMT-based quantitative proteomics reveals the nutritional and stress resistance functions of anaerobic fungi in yak rumen during passage at different time intervals.

OBJECTIVES: Anaerobic fungi are critical for nutrient digestion in the yak rumen. Although studies have reported the effects of passage at different time intervals on the community structure of yak rumen anaerobic fungi, it is unknown whether passage culture at different time intervals affects the microbial proteins of rumen anaerobic fungi and their functions. METHODS: Mycelium was obtained using the anaerobic continuous batch culture (CBC) of yak rumen fluid at intervals of 3 d, 5 d and 7 d. Quantitative analysis of fungal proteins and functional analysis was performed using tandem mass tagging (TMT) and bioinformatics. RESULTS: A total of 56 differential proteins (DPs) were found in 5 d vs. 3 d and 7 d vs. 3 d. Gene ontology (GO) enrichment indicated that the up-regulated proteins were mainly involved in biological regulation, cellular process, metabolic process, macromolecular complex, membrane, cell part, organelle, binding, catalytic activity and transporter activity. The downregulated proteins were mainly enriched in metabolic process, cell part, binding and catalytic activity. Furthermore, the downregulated proteins in 7 d vs. 3 d were related to membrane and organelle. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment results indicated that DPs were enriched in 14 pathways in 5 d vs. 3 d and 7 d vs. 3 d, mainly including terpenoid backbone biosynthesis, alaine, aspartate and glutamate metabolism, arginine biosynthesis, hypotaurine, cyanoamino acid, glutathione, ß-alanine, pyrimidine, purine, galactose and propanate metabolism, steroid biosynthesis, ribosome biogenesis in eukaryotes and aminoacyl tRNA biosynthesis. The DPs were enriched in only 2 pathways in 5 d vs 3 d, lysine biosynthesis and cysteine and methionine metabolism. N-glycan biosynthesis and retinol metabolism are only found in the metabolism of DPs in 7 d vs 3 d. CONCLUSIONS: Yak rumen anaerobic fungal proteins are involved in nutrition and stress tolerance during passage at different time intervals.

Effects of water saving and nitrogen reduction on the yield, quality, water and nitrogen use efficiency of Isatis indigotica in Hexi Oasis.

Isatis indigotica planting is the backbone of the medicinal industry in Hexi Oasis, Gansu. In order to solve the problems insufficient water resources and excessive application of nitrogen fertilizer in this area, this paper explored the irrigation and nitrogen levels that can meet the multiple goals of Isatis indigotica. The two-factor split-plot field experiment (2018‒2019) was conducted in Minle County, Gansu Province, China, which contains 9 treatments. There were three levels of irrigation water: W1(low), W2(medium), and W3(high). The soil moisture contents were 60-70%, 70-80%, and 80-90% of the field water-holding capacity, respectively. The nitrogen application rate was classified into three levels, N1(low), N2(medium) and N3(high), which were 150, 200 and 250 kg N/ha, respectively. The standard local irrigation water amount and nitrogen application rate corresponded to W3N3. The results showed that the yield of Isatis indigotica increased first and then decreased with the increase of irrigation amount and nitrogen application rate, the yield of W2N2 is 12.2-17.1% higher than that of W1N1, the yield of W3N3 was 12.1-17.5% lower than that of W2N2. Saving water and reducing nitrogen can improve the quality of Isatis indigotica, compared with W3N3, the indigo, indirubin, (R,S)-epigoitrin and polysaccharides of W2N2 increased by 4.5-5.9%, 2.7-3.1%, 5.2-6.0%, and 1.8-2.1%, respectively. With the increase of nitrogen application rate, the water use efficiency (WUE) first increased and then decreased, as the irrigation volume increases, WUE decreases. Compared with W3N3, the WUE of W2N2 increased by 24.3-27.2%. With the increase of water input, the nitrogen fertilizer use efficiency (NUE) first increased and then decreased, as the nitrogen application rate increases, NUE decreases. Compared with W3N3, the NUE of W2W2 increased by 31.8-34.5%. Therefore, W2N2 can improve quality and increase water and nitrogen utilization efficiency on the basis of ensuring yield.

Redescription and phylogenetic position of the terrestrial ciliates Gastrostylides dorsicirratus and Heterourosomoida lanceolata (Hypotricha, Dorsomarginalia).

The morphology and infraciliature of two soil hypotrichous ciliates, Heterourosomoida lanceolata (Shibuya, 1930) Singh and Kamra, 2015 and Gastrostylides dorsicirratus (Foissner, 1982) Foissner, 2016, were investigated using live observation and protargol staining. The Chinese population of H. lanceolata differs slightly from other populations in the body size in vivo, the relative length of the adoral zone, the number of right and left marginal cirri, the total number of dorsal bristles, and the number of micronuclei. The Chinese population of G. dorsicirratus corresponds well with Austrian and Indian populations. We also document the morphogenetic processes during binary fission of G. dorsicirratus, including the formation of the frontoventral-transverse cirral anlagen in a primary pattern. In addition, phylogenetic analyses based on SSU rDNA sequence data reveal that the two populations of G. dorsicirratus for which data are available cluster together with full support and form a clade with Heterourosomoida sinica and two populations of H. lanceolata.

Effects of swine manure composting by microbial inoculation: Heavy metal fractions, humic substances, and bacterial community metabolism.

This study aimed to investigate the effect of swine manure composting with microbial inoculation (MI) and without MI (CK) on heavy metal (Cr, Cd, and Pb) fractions, humic substance (HS), and metabolism pathway. The results showed that MI could passivate the heavy metal Cr and reduce the proportion of exchangeable (EXC) fraction of Cd, but it does not affect the EXC fraction of Pb. Compared to CK, HS, humic acid (HA), and fulvic acid (FA) were significantly increased with MI at the maturity stage. The propagation of Proteobacteria (day 4) and Firmicutes (days 12 and 24) was strengthened with MI. Canonical correlation analysis found that HA and Firmicutes were positively correlated with heavy metal (Cr, Cd, and Pb) residual (RES) fraction, and FA was positively correlated with Proteobacteria. Moreover, MI can significantly increase amino acid metabolism and carbohydrate metabolism by day 4, enhance the metabolism of enzyme families and glycan biosynthesis by day 12, and improve membrane transport. Overall, MI could facilitates the increase in HA and FA content and transfer of heavy metal (Cr, Cd, and Pb) fractions, it particularly helps increase the RES fraction.

Glucose-induced changes in the bacterial communities of mine tailings at different acidification stages.

Ecological restoration technologies applied to tailings can influence the associated bacterial communities. However, it is unknown if the shifts in these bacterial communities are caused by increased organic carbon. Glucose-induced respiration and high-throughput sequencing were used to assess the microbial activity and bacterial communities, respectively. Glucose addition increased the microbial activity, and glucose + ammonium nitrate addition resulted in slightly higher CO2 emission than did glucose addition alone, suggesting that carbon and nitrogen limited microbial community growth. In neutral pH tailings, the bacterial taxa that increased by glucose addition were assigned to the phyla Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Planctomycetes. However, the bacterial taxa that increased by glucose addition in acidic tailings only belonged to the phylum Actinobacteria (maximum increase of 43.78%). In addition, the abundances of the total nitrogen-fixing genera and of the genus Arthrobacter (representing approximately 97.89% of the total nitrogen-fixing genera) increased by glucose addition in acidic tailings (maximum increase of 46.98%). In contrast, the relative abundances of the total iron- and (or) sulfur-oxidizing bacteria decreased (maximum decrease of 10.41%) in response to the addition of glucose. These findings indicate that the addition of organic carbon is beneficial to the development of bacterial communities in mine tailings.

Dietary intakes and dietary patterns among pregnant women in Northwest China.

OBJECTIVE: To describe nutrient intakes, characterize dietary patterns and analyse their associations with sociodemographic characteristics among pregnant women in Shaanxi, China. DESIGN: Population-based cross-sectional survey. SETTING: Twenty counties and ten districts in Shaanxi Province of Northwest China, 2013. SUBJECTS: Women (n 7462) were recruited using a stratified multistage random sampling method to report diets during pregnancy, at 0-12 months (median 3 months; 10th-90th percentile, 0-7 months) after delivery. RESULTS: Pregnant women had higher intakes of fat, niacin and vitamin E than the nutrient reference values, while most micronutrients such as vitamin A, folate, Ca and Zn were reportedly low. Women in the highest education, occupation and household income groups had higher nutrient intakes than those in the lowest groups. Nutrient intake differences also existed by geographic area, residence and maternal age at delivery. Three dietary patterns were identified: balanced pattern, vegetarian pattern and snacks pattern. Participants with high balanced pattern scores tended to be better educated, wealthier, 25-29 years old at delivery, working outside and living in urban areas and central Shaanxi. Women with high scores on the vegetarian pattern and snacks pattern tended to be in low balanced pattern score groups, and had lower nutrient intakes than those in the high balanced pattern score groups. CONCLUSIONS: The study suggested that pregnant women in Shaanxi, China had low intakes of most nutrients such as vitamin A, folate and Ca. Dietary patterns and most nutrient intakes varied by sociodemographic characteristics. Targeted programmes are needed to improve dietary intakes and dietary patterns among sociodemographically disadvantaged groups.

3-Methylthiopropionic Acid of Rhizoctonia solani AG-3 and Its Role in the Pathogenicity of the Fungus.

Studies were conducted to determine the role of 3-methylthioproprionic acid (MTPA) in the pathogenicity of potato stem canker, Rhizoctonia solani, and the concentrations required to inhibit growth of R. solani under laboratory and plant house-based conditions. The experiments were laid out in a completely randomized design with five treatments and five replications. The treatments were 0, 1, 2, 4, and 8 mM concentrations of MTPA. The purified toxin exhibited maximal activity at pH 2.5 and 30°C. MTPA at 1, 2, 4, and 8 mM levels reduced plant height, chlorophyll content, haulm fresh weight, number of stolons, canopy development, and tuber weight of potato plants, as compared to the control. MTPA significantly affected mycelial growth with 8 mM causing the highest infection. The potato seedlings treated with MTPA concentrations of 1.0-8.0 mM induced necrosis of up to 80% of root system area. Cankers were resulted from the injection of potato seedling stems with 8.0 mM MTPA. The results showed the disappearance of cell membrane, rough mitochondrial and cell walls, change of the shape of chloroplasts, and swollen endoplasmic reticulum. Seventy-six (76) hours after toxin treatment, cell contents were completely broken, cytoplasm dissolved, and more chromatin were seen in the nucleus. The results suggested that high levels of the toxin concentration caused cell membrane and cytoplasm fracture. The integrity of cellular structure was destroyed by the phytotoxin. The concentrations of the phytotoxin were significantly correlated with pathogenicity and caused damage to the cell membrane of potato stem base tissue.

[Control of continuous potato monoculture barrier via biological soil disinfestation method in Yellow River irrigation areas of central Gansu Province, Northwest China].

The potential of biological soil disinfestation (BSD) in control of continuous potato monoculture barrier was investigated in present study. BSD involves the induction of soil reduction conditions through incorporation of easily decomposed organic materials into soil, flooding the soil by irrigation, and covering the soil surface with plastic film. Control (CK) was left without cover and organic amendment as well as flooding. Field experiment was conducted for testing the effect of BSD approach on the control of continuous potato monoculture barrier, especially on tube yield, plant growth and development, suppression of soil-borne pathogen, and soil microbial community and enzyme activities. Compared with CK, BSD treatment significantly increased tuber yield by 16.1% and plant biomass by 30.8%, respectively. Meanwhile, the incidence of diseased plant and the ratio of diseased tuber in BSD treatment also significantly decreased by 68.0% and 46.7% as compared to those in CK, respectively. BSD treatment significantly increased the content of chlorophyll and branch numbers per main stem of potato plants, improved the morphological characteristics of potato root system. In the course of BSD before potato sowing, soil pH value and bacteria/fungi significantly increased, but populations of fungi and Fusarium sp. significantly decreased compared with CK. There were no significant changes in populations of bacteria and actinomycetes between CK and BSD treatments. During potato growing stage, the populations of both soil fungi and Fusarium sp. were lower in BSD treatment than those of CK. With the advance of potato growth, the population of Fusarium sp. in BSD treatment gradually increased compared with CK. There were no significant changes in soil enzyme activities in the course of BSD before potato sowing and the whole of potato growing stage. It was concluded that BSD has the potential to control continuous potato monoculture barrier and may be an important element in a sustainable and effective management strategy for potato soil-borne diseases.

[Identification of chemicals in root exudates of potato and their effects on Rhizoctonia solani].

This study was conducted to identify chemicals in root exudates and their effect on Rhizoctonia solani in potato cropping systems. Root exudates were collected from the fields with 5 years of continuous potato cropping in comparison with rotational cropping of potato and other crops, using in-house made root boxes at the seedling and squaring stages. Chemicals in the root exudates were identified using the GC-MS method. The results showed that glucide concentration was the highest in the root exudates, followed by organic acids. Compared with the rotational cropping, the continuous cropping significantly decreased the glucide content and increased the content of organic acids in the root exudates. The contents of almitic acid in root exudates under continuous cropping was 0.94% at seedling stage and 1.4% at squaring stage, the dibutyl phthalate was 0.15%, whereas under rotational cropping, those values were decreased to 0.15%, 0.2%, and being negligible, respectively. The root exudates promoted the growth of R. solani, especially under continuous potato cropping. The simulation test showed that the palmitic acid and dibutyl phthalate in root exudates could promote the growth of R. solani.

Prenatal Alcohol Exposure and Congenital Heart Defects: A Meta-Analysis.

BACKGROUND: There are still inconsistent conclusions about the association of prenatal alcohol drinking with congenital heart defects (CHDs). We conducted this meta-analysis to investigate the association between prenatal alcohol exposure and the risk of overall CHDs and the CHDs subtypes. METHODS: Case-control and cohort studies published before March 2015 were searched through PubMed and Embase. Two authors independently extracted data and scored the study quality according to the Newcastle-0ttawa Scale. The pooled ORs and 95%CI were estimated using the random-effects model and heterogeneity was assessed by the Q test and I2 statistic. RESULTS: A total of 20 studies were finally included. The results provided no evidence of the association between prenatal alcohol exposure and the risk of overall CHDs (OR = 1.06, 95%CI = 0.93-1.22), ventricular septal defects (VSDs) (OR = 1.04, 95%CI = 0.86-1.25), or atrial septal defects (ASDs) (OR = 1.40, 95%CI = 0.88-2.23). However, prenatal alcohol drinking was marginally significantly associated with conotruncal defects (CTDs) (OR = 1.24, 95%CI = 0.97-1.59) and statistically significantly associated with d-Transposition of the Great Arteries (dTGA) (OR = 1.64, 95%CI = 1.17-2.30). Moreover, both prenatal heavy drinking and binge drinking have a strong association with overall CHDs (heavy drinking: OR = 3.76, 95%CI = 1.00-14.10; binge drinking: OR = 2.49, 95%CI = 1.04-5.97), and prenatal moderate drinking has a modest association with CTDs (OR = 1.35, 95%CI = 1.05-1.75) and dTGA (OR = 1.86, 95%CI = 1.09-3.20). CONCLUSIONS: In conclusion, the results suggested that prenatal alcohol exposure was not associated with overall CHDs or some subtypes, whereas marginally significant association was found for CTDs and statistically significant association was found for dTGA. Further prospective studies with large population and better designs are needed to explore the association of prenatal alcohol exposure with CHDs including the subtypes in specific groups.

Microbial community diversities and taxa abundances in soils along a seven-year gradient of potato monoculture using high throughput pyrosequencing approach.

BACKGROUND: Previous studies have focused on linking soil community structure, diversity, or specific taxa to disturbances. Relatively little attention has been directed to crop monoculture soils, particularly potato monoculture. Information about microbial community changes over time between monoculture and non-monoculture treatments is lacking. Furthermore, few studies have examined microbial communities in potato monoculture soils using a high throughput pyrosequencing approach. METHODOLOGY/PRINCIPAL FINDINGS: Soils along a seven-year gradient of potato monoculture were collected and microbial communities were characterized using high throughput pyrosequencing approach. Principal findings are as follows. First, diversity (H(Shannon)) and richness (S(Chao1)) indices of bacterial community, but not of fungal community, were linearly decreased over time and corresponded to a decline of soil sustainability represented by yield decline and disease incidence increase. Second, Fusarium, the only soilborne pathogen-associated fungal genus substantially detected, was linearly increased over time in abundance and was closely associated with yield decline. Third, Fusarium abundance was negatively correlated with soil organic matter (OM) and total nitrogen (TN) but positively with electrical conductivity (EC). Fourth, Fusarium was correlated in abundances with 6 bacterial taxa over time. CONCLUSIONS: Soil bacterial and fungal communities exhibited differential responses to the potato monoculture. The overall soil bacterial communities were shaped by potato monoculture. Fusarium was the only soilborne pathogen-associated genus associated with disease incidence increase and yield decline. The changes of soil OM, TN and EC were responsible for Fusarium enrichment, in addition to selections by the monoculture crop. Acidobacteria and Nitrospirae were linearly decreased over time in abundance, corresponding to the decrease of OM, suggesting their similar ecophysiologial trait. Correlations between abundance of Fusarium with several other bacterial taxa suggested their similar behaviors in responses to potato monoculture and/or soil variables, providing insights into the ecological behaviors of these taxa in the environment.

[Characteristics of dry matter and potassium accumulation and distribution in potato plant in semi-arid rainfed areas].

In 2010, a field experiment with potato (Solanum tuberosum) cultivar 'Xindaping' was conducted at the Dingxi Extension Center of Gansu Province, Northwest China, aimed to understand the accumulation and distribution patterns of dry matter (DM) and potassium (K) in the organs of potato plant in semi-arid rainfed areas. During the whole growth period of the cultivar, the DM accumulation in root, stem, and leaf all showed a unimodal curve, with the DM accumulation rate being leaf > stem > root, whereas the DM accumulation in whole plant and tuber was an S-curve. The maximum DM accumulation rate of the whole plant was higher than that of the tuber, and appeared 17 days earlier. The distribution of DM in different organs showed two turning points, i.e., during the tuber formation (TF) period and the tuber growth (TG) period. During TF period, the DM accumulation was the greatest in leaf, followed by in tuber. The TF period was also the DM balance period, which occurred 90 days after emergence. Before the DM balance period, the DM accumulation in tuber was lesser than that in root, stem, and leaf, and there was a positive correlation between the DM accumulation in tuber and in root, stem, and leaf. However, after the DM balance period, the DM accumulation in tuber was greater than that in root, stem, and leaf, and the correlation was negative. At seedling stage and in TF period, TG period, starch accumulation period, and maturity period, the DM accumulation in whole plant was 5%, 30%, 60%, 4% , and 1%, while that in tuber was 0,18% , 62 , 18% , and 2%, respectively. In the whole growth period, more than 50% of the DM was formed in TG period. The K concentration was the highest in stem and the lowest in tuber, though the K was mostly concentrated in root before the DM balance period. The K accumulation before the DM balance period was mostly in root, stem, and leaf, with the sequence of stem > leaf > root, but after the DM balance period, the K was mainly allocated in tuber, with >60% of the K accumulated in tuber in maturity period.

[Fungal population structure and its biological effect in rhizosphere soil of continuously cropped potato].

Continuous cropping obstacle is one of the main restriction factors in potato industry. In order to explore the mechanisms of potato's continuous cropping obstacle and to reduce the impact on potato's tuber yield, a field experiment combined with PCR-DGGE molecular fingerprinting was conducted to investigate the fungal population structure and its biological effect in rhizosphere soil of continuously cropped potato. With the increasing year of potato' s continuous cropping, the numbers of visible bands in rhizosphere fungal DGGE profiles increased obviously. As compared with that of CK (rotation cropping), the operational taxonomic unit (OTU) in treatments of one to five years continuous cropping was increased by 38.5%, 38.5%, 30.8%, 46.2%, and 76.9% respectively, indicating that potato's continuous cropping caused an obvious increase in the individual numbers of dominant fungal populations in rhizosphere soil. Also with the increasing year of potato's continuous cropping, the similarity of the fungal population structure among the treatments had a gradual decrease. The sequencing of the fungal DGGE bands showed that with the increasing year of continuous cropping, the numbers of the potato's rhizosphere soil-borne pathogens Fusarium oxysporum and F. solani increased obviously, while the number of Chaetomium globosum, as a biocontrol species, had a marked decrease in the fifth year of continuous cropping. It was suggested that potato' s continuous cropping caused the pathogen fungal populations become the dominant microbial populations in rhizosphere soil, and the rhizosphere micro-ecological environment deteriorated, which in turn affected the root system, making the root vigor and its absorption area reduced, and ultimately, the tuber yield decreased markedly.