Using a DNA mini-barcode within the ITS region to identify toxic Amanita in mushroom poisoning cases.

Mushroom poisoning contributes significantly to global foodborne diseases and related fatalities. Amanita mushrooms frequently cause such poisonings; however, identifying these toxic species is challenging due to the unavailability of fresh and intact samples. It is often necessary to analyze residues, vomitus, or stomach extracts to obtain DNA sequences for the identification of species responsible for causing food poisoning. This usually proves challenging to obtain usable DNA sequences that can be analyzed using conventional molecular biology techniques. Therefore, this study aimed to develop a DNA mini-barcoding method for the identification of Amanita species. Following the evaluation and optimization of universal primers for DNA mini-barcoding in Amanita mushrooms, we found that the internal transcribed spacer (ITS) gene sequence primer ITS-a was the most suitable DNA barcode primer for identifying Amanita species. Forty-three Amanita samples were subsequently amplified and sequenced. The sequences obtained were analyzed for intra- and inter-species genetic distances, and a phylogenetic tree was constructed. The findings indicated that the designed primers had strong universality among the Amanita samples and could accurately identify the target gene fragment with a length of 290 bp. Notably, the DNA mini-barcode accurately identified the 43 Amanita samples, demonstrating high consistency with the conventional DNA barcode. Furthermore, it effectively identified DNA from digested samples. In summary, this DNA mini-barcode is a promising tool for detecting accidental ingestion of toxic Amanita mushrooms. It may be used as an optimal barcode for species identification and traceability in events of Amanita-induced mushroom poisoning. KEY POINTS: • Development of a DNA mini-barcoding method for Amanita species identification without fresh samples. • The ITS-a primer set was optimized for robust universality in Amanita samples. • The mini-barcode is suitable for screening toxic mushroom species in mushroom poisoning cases.

[Effects of Stimulated Nitrogen Deposition on the Bacterial Community Structure of Semiarid Temperate Grassland].

With the development of economics, the deposition of available nitrogen in the terrestrial ecosystem is increasing dramatically due to anthropic activities, which negatively impacts the sustainability of the ecosystem ecology. In this study, the effect of long-term stimulated nitrogen deposition[with nitrogen addition of 0, 1, 4, 8, and 32 g·(m2·a)-1] on the microbial community structure of soil was investigated in a temperate steppe in Inner Mongolia using a pyrosequencing technique targeting the bacterial 16S rRNA gene. The results show that the available nitrogen in soil increases with increasing nitrogen addition, resulting in the decrease of the soil pH. The results of pyrosequencing indicate that soil bacterial OTU (operational taxonomy unit) numbers increase with increasing nitrogen deposition, while bacterial α diversity indices show an initial increase and subsequent decrease. Non-metric multidimensional scaling (NMDS) analysis indicates that the bacterial community structure significantly varies among treatments, which can be largely attributed to the changes in the soil pH and nitrogen content due to nitrogen deposition. At the class level, the relative abundance of different bacterial groups shows a varying trend depending on the nitrogen deposition. This study indicates that long-term nitrogen deposition significantly impacts the bacterial community by changing the soil properties.

Exogenous N addition enhances the responses of gross primary productivity to individual precipitation events in a temperate grassland.

Predicted future shifts in the magnitude and frequency (larger but fewer) of precipitation events and enhanced nitrogen (N) deposition may interact to affect grassland productivity, but the effects of N enrichment on the productivity response to individual precipitation events remain unclear. In this study, we quantified the effects of N addition on the response patterns of gross primary productivity (GPP) to individual precipitation events of different sizes (Psize) in a temperate grassland in China. The results showed that N enrichment significantly increased the time-integrated amount of GPP in response to an individual precipitation event (GPPtotal), and the N-induced stimulation of GPP increased with increasing Psize. N enrichment rarely affected the duration of the GPP response, but it significantly stimulated the maximum absolute GPP response. Higher foliar N content might play an important role in the N-induced stimulation of GPP. GPPtotal in both the N-addition and control treatments increased linearly with Psize with similar Psize intercepts (approximately 5 mm, indicating a similar lower Psize threshold to stimulate the GPP response) but had a steeper slope under N addition. Our work indicates that the projected larger precipitation events will stimulate grassland productivity, and this stimulation might be amplified by increasing N deposition.

[Influence of nitrogen and phosphorus addition on the aboveground biomass in Inner Mongo- lia temperate steppe, China].

The plants in arid environment are constrained not only by water availability, but also by soil nutrient conditions. In order to clarify to what extent nutrient addition would facilitate the growth of plants in semi-arid region, we conducted a nitrogen (N) and phosphorus (P) addition experiment in Inner Mongolia temperate grassland in 2012 and 2013. In our experiment, N was added at 10 and 40 g N · m(-2) · a(-1) alone or in combination with P addition (10 g P · m(-2) · a(-1)). N addition significantly improved plant aboveground biomass (AGB) during the two study years. AGB in the treatments of 10 and 40 g · m2 · a(-1) was enhanced by 50.8% and 65.9% in 2012, and 71.6% and 93.3% in 2013, respectively. However, no significant difference in AGB enhancement was found between two N addition treatments. Compared with N addition treatments at the rates of 10 and 40 g · m(-2) · a(-1), N plus P addition improved AGB by 98.4% and 186.8% in 2012, and 111.7% and 141.4% in 2013, respectively. N addition generally increased all the three main functional types (i.e., Gramineae, Asteraceae and others) , and the three functional types contributed nearly equally to the increase of the community AGB. In comparison, Asteraceae contributed largest to the increments of AGB under the N plus P addition treatments. Our results also indicated that N and P addition remarkably increased the ground coverage, resulting in improved surface soil moisture condition, which might be one important reason that N and P addition could facilitate plant growth in arid environment.

[Comparative study of biological characters and paeoniflorin content of wild and asexual cultivated Paeonia lactiflora growing at Duolun county, Inner Mongolia].

This study aims to investigate whether the cultivation peony, can take the place of wild herbaceous peony by comparing the biological traits and paeoniflorin content between them. The result showed that the biomass of the stem, leaf, crown, fleshy root and fine root of wild plants were all smaller than that of bud asexual cultivated plants, while there was no significant differences in below-ground and aboveground biomass ratio between these two plants. The stele diameter, the proportion of stele, and the ratio of stele diameter to cortex thickness of wild plants were significantly higher than that of bud asexual cultivated plants, while the cortex thickness and the proportion of cortex were significantly smaller than bud asexual cultivated plants. Although the biological traits of bud asexual cultivated plants have changed significantly, the paeoniflorin content in fleshy roots has no significant difference between wild and bud asexual cultivated plants. Therefore, it is feasible to use the bud asexual cultivation to the conservation and large-scale cultivation of Paeonia laciflora, which is an endangered species.

Nitrogen response efficiency increased monotonically with decreasing soil resource availability: a case study from a semiarid grassland in northern China.

The concept of nutrient use efficiency is central to understanding ecosystem functioning because it is the step in which plants can influence the return of nutrients to the soil pool and the quality of the litter. Theory suggests that nutrient efficiency increases unimodally with declining soil resources, but this has not been tested empirically for N and water in grassland ecosystems, where plant growth in these ecosystems is generally thought to be limited by soil N and moisture. In this paper, we tested the N uptake and the N use efficiency (NUE) of two Stipa species (S. grandis and S. krylovii) from 20 sites in the Inner Mongolia grassland by measuring the N content of net primary productivity (NPP). NUE is defined as the total net primary production per unit N absorbed. We further distinguished NUE from N response efficiency (NRE; production per unit N available). We found that NPP increased with soil N and water availability. Efficiency of whole-plant N use, uptake, and response increased monotonically with decreasing soil N and water, being higher on infertile (dry) habitats than on fertile (wet) habitats. We further considered NUE as the product of the N productivity (NP the rate of biomass increase per unit N in the plant) and the mean residence time (MRT; the ratio between the average N pool and the annual N uptake or loss). The NP and NUE of S. grandis growing usually in dry and N-poor habitats exceeded those of S. krylovii abundant in wet and N-rich habitats. NUE differed among sites, and was often affected by the evolutionary trade-off between NP and MRT, where plants and communities had adapted in a way to maximize either NP or MRT, but not both concurrently. Soil N availability and moisture influenced the community-level N uptake efficiency and ultimately the NRE, though the response to N was dependent on the plant community examined. These results show that soil N and water had exerted a great impact on the N efficiency in Stipa species. The intraspecific differences in N efficiency within both Stipa species along soil resource availability gradient may explain the differences in plant productivity on various soils, which will be conducive to our general understanding of the N cycling and vegetation dynamics in northern Chinese grasslands.