高含固率木质纤维素厌氧发酵物总DNA的4种提取方法比较

不同的样本特性和提取方法对获得微生物总DNA的质量有重要影响。文章基于高含固率木质纤维素厌氧发酵物腐殖酸、酚类物质含量高、质地均一性差、微生物浓度低的特点,研究了4种方法提取不同高含固率粪秸厌氧发酵物中微生物总DNA的效果。结果表明,常规的十二烷基磺酸钠法(sodium dodecyl sulfate,SDS)、十二烷基磺酸钠和溴化十六烷基三甲铵结合法(sodium dodecyl sulfate and cetyltrimethyl ammonium bromide,SDS-CTAB)和商业的粪便试剂盒法提取的DNA质量均较差,SDS法和试剂盒法未能获得聚合酶链式反应(polymerase chain reaction,PCR)扩增目的条带,SDS-CTAB法得到的条带较模糊;改进SDS-CTAB法获得的DNA杂质少、纯度高,具有较好的稳定性,A260/A280和A260/A230值分别为1.74~1.86和1.65~1.86,每克样品的DNA浓度在50 ng·μL^-1以上,电泳条带单一齐整、清晰明亮,PCR扩增的目的条带清晰度高,适宜后续分子生物学技术的分析。林格氏液洗脱、聚乙烯吡咯烷酮-40(Polyvinyl Pyrrolidone-40,PVP-40)洗涤液除杂以及裂解液和多种酶联合破壁是改进SDS-CTAB法获得该类专一性样本高质量微生物总DNA的关键步骤。     

Does sand content in spawning substrate result in early larval emergence? Evidence from a lithophilic cyprinid fish

The spawning success of lithophilic salmonids is strongly influenced by the fine sediment content (“fines”) of spawning substrates, yet knowledge on the impacts of fines on the spawning of non‐salmonid lithophiles remains limited, despite their ecological and socio‐economic importance in European rivers. Consequently, the aim here was to use an ex‐situ experiment to investigate the impact of sand content on egg survival and timing of larval emergence of the surface‐spawning cyprinid European barbel Barbus barbus. Thirty incubator boxes within a recirculating system were filled with one of five experimental sediment mixtures (0%–40% sand by mass) that each contained 300 fertilised eggs at a depth of 50 mm. Emerged, free‐swimming larvae were captured and counted daily to assess grain‐size effects on larval survival and emergence. Specifically, total proportion of emerged larvae, cumulative daily proportion of emerged larvae and time required to reach 50% emergence were measured during the study. Whilst the proportion of sand in the sediments did not have a significant impact on egg‐to‐emergence survival (mean survival per treatment 75%–79%), it significantly affected the timing of larval emergence to the water column; early emergence was detected in treatments with elevated sand content (on average, 50% emergence after 12–13 days versus 19 days in the control). Similar to findings from salmonid studies, these results suggest high sand content in spawning gravels can influence timing of larval emergence and potentially cyprinid lithophilic fish survival.     

Carbon stocks and carbon fluxes from a 10‐year prescribed burning chronosequence on a UK blanket peat

Prescribed burning is a common land management technique in many areas of the UK uplands. However, concern has been expressed at the impact of this management practice on carbon stocks and fluxes found in the carbon‐rich peat soils that underlie many of these areas. This study measured both carbon stocks and carbon fluxes from a chronosequence of prescribed burn sites in northern England. A range of carbon parameters were measured including above ground biomass and carbon stocks; net ecosystem exchange (NEE), net ecosystem respiration (R_eco) and photosynthesis (P_g) from closed chamber methods; and particulate organic carbon (POC). Analysis of the CO₂ data showed that burning was a significant factor in measured CO₂ readings but that other factors such as month of sampling explained a greater proportion of the variation in the data. Carbon budget results showed that whereas all the plots were net sources of carbon, the most recent burn scars were smaller sources of carbon compared with the older burn scars, suggesting that burning of Calluna‐dominated landscapes leads to an ‘avoided loss’ of carbon. However, this management intervention did not lead to a transition to a carbon sink and that for carbon purposes, active peat‐forming conditions are desirable.     

Greater humification of belowground than aboveground biomass carbon into particulate soil organic matter in no-till corn and soybean crops

Quantifying the amount of carbon (C) incorporated from decomposing residues into soil organic carbon (C_S) requires knowing the rate of C stabilization (humification rate) into different soil organic matter pools. However, the differential humification rates of C derived from belowground and aboveground biomass into C_S pools has been poorly quantified. We estimated the contribution of aboveground and belowground biomass to the formation of C_S in four agricultural treatments by measuring changes in δ¹³C natural abundance in particulate organic matter (C_POM) associated with manipulations of C₃ and C₄ biomass. The treatments were (1) continuous corn cropping (C₄ plant), (2) continuous soybean cropping (C₃), and two stubble exchange treatments (3 and 4) where the aboveground biomass left after the grain harvest was exchanged between corn and soybean plots, allowing the separation of aboveground and belowground C inputs to C_S based on the different δ¹³C signatures. After two growing seasons, C_POM was primarily derived from belowground C inputs, even though they represented only ∼10% of the total plant C inputs as residues. Belowground biomass contributed from 60% to almost 80% of the total new C present in the C_POM in the top 10 cm of soil. The humification rate of belowground C inputs into C_POM was 24% and 10%, while that of aboveground C inputs was only 0.5% and 1.0% for soybean and corn, respectively. Our results indicate that roots can play a disproportionately important role in the C_POM budget in soils. Keywords Particulate organic matter; root carbon inputs; carbon isotopes; humification rate; corn; soybean.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

Indicators and trade-offs of ecosystem services in agricultural soils along a landscape heterogeneity gradient

Soil functions can be classified as supporting (nutrient cycling) and provisioning (crop production) ecosystem services (ES). These services consist of multiple and dynamic functions and are typically assessed using indicators, e.g. microbial biomass as an indicator of supporting services. Agricultural intensification negatively affects indicators of soil functions and is therefore considered to deplete soil ES. It has been suggested that incorporating leys into crop rotations can enhance soil ES. We examined this by comparing indicators of supporting soil services – organic carbon, nitrogen, water holding capacity and available phosphorous (carbon storage and nutrient retention); net nitrogen mineralisation rate and microbial biomass (nutrient cycling and retention) – in barley fields, leys and permanent pastures along a landscape heterogeneity gradient (100, 500 and 1000 m radii). In addition, barley yields (provisioning service) were analysed against these indicators to identify trade-offs among soil services. Levels of most indicators did not differ between barley and ley fields and were consistently lower than in permanent pastures. Leys supported greater microbial biomass than barley fields. Landscape heterogeneity had no effect on the indicators or microbial community composition. However, landscape heterogeneity correlated negatively with yield and soil pH, suggesting that soils in heterogeneous landscapes are less fertile and therefore have lower yields. No trade-offs were found between increasing barley yield and the soil indicators. The results suggest that soil ES are determined at the field level, with little influence from the surrounding landscape, and that greater crop yields do not necessarily come at the expense of supporting soil services.     

微波辐射法竹浆纤维素的中性TEMPO氧化及表征

采用微波辐射法和中性2,2,6,6-四甲基哌啶-1-氧化物(TEMPO)自由基氧化工艺来预处理竹浆纤维素,之后用超声波机械处理制备微纤化纤维素。实验结果表明,在60℃下,TEMPO用量为0.15 mmol/g(以绝干浆计,下同),Na Cl O2用量为10 mmol/g,Na Cl O用量为1 mmol/g,微波氧化2 h,其羧基含量最高到达0.729 mmol/g,氧化效果最佳,超声波处理后微纤化纤维素得率高达85.7%。FT-IR分析表明纤维素氧化后引入羧基官能团;XRD结果揭示纤维素氧化反应只发生在纤维素的无定型区或结晶区表面;纤维素氧化前后的聚合度(Dp)结果表明相对于碱性氧化,微波辐射和中性TEMPO氧化对纤维的降解程度低,从而有效地保持了纤维素原本的长度。     

Cross‐linked polyacrylates in post‐mining substrates: persistence and effects on plant growth

Application of hydrophilic polymers composed of cross‐linked polyacrylate can improve soil water‐holding capacity and accelerate the restoration of post‐mining substrates. In this work, we studied the persistence of a polyacrylate polymer incorporated into a soil and its impact on plant nutrients at a reclamation site of former lignite mining in Lusatia (Germany). In contrast to autumn application, the incorporation of the polymer enhanced the sequestration of plant‐derived carbon in the soil, which was reflected by a significant increase in the concentration of a lignin marker. Attenuated total reflexion–Fourier transform infrared spectra (ATR‐FTIR) and total elemental contents in the applied polymer suggested an intensive cation exchange between the polymer framework and the soil‐forming substrate. In addition, there was an enrichment of carbonaceous material, which seems to reduce the swelling and thus the water‐holding capacity of the cross‐linked polyacrylate. Conversely, this process protected the polymer structure from rapid decomposition.     

云南传统水稻品种W axy基因多样性形成和维持机制初探

本研究针对W axy基因第四外显子和第五外显子的3个功能位点开发了新的AS-PCR标记，完善了W axy基因的分子标记体系，为更精确的鉴定和筛选目标W axy等位基因提供了工具。直链淀粉含量测定和AS-PCR分析结果显示，云南传统水稻品种具有极为丰富的W axy基因多样性和直链淀粉含量变异，308个传统品种中共检测到65种不同的W axy单倍型，直链淀粉含量变幅为0~48%。In1、E6、E10和E4op/hp这4个位点的碱基频率在四大稻区间存在显著差异，E2位点和In1位点的碱基频率在不同民族间存在显著差异，表明不同稻作区、不同民族对水稻直链淀粉含量和W axy基因有着多样化的选择。同时民族混居地区E2和In1位点的碱基频率不同于其相应的民族独居地区，说明不同民族混居也会产生对W axy基因不同的选择并增加W axy基因的多样性。多样化的选择和民族混居是云南传统水稻品种W axy基因多样性形成和维持的重要机制。在云南四大稻作区中，南部边缘水陆稻区的W axy单倍型最为丰富，共检测到50种单倍型，占总数的77%，这一区域应被设为云南传统稻种资源的保护优先区和核心区。     

Physicochemical characterization and in-vitro digestibility of extruded rice noodles with different amylose contents based on rheological approaches

The physicochemical properties and in-vitro digestibility of extruded rice noodles with different amylose contents were characterized from a rheological point of view. Thermo-mechanical measurements showed that the rice flour with higher amylose contents exhibited greater stability to dual-mixing and higher degrees of starch gelatinization and retrogradation. In addition, greater elastic properties were clearly observed in the high amylose rice samples. The use of high amylose rice flour produced noodles with a harder texture, consequently contributing to reduced cooking loss. Furthermore, the rheological changes of extruded rice noodles were monitored in real time during the in-vitro starch digestion. The rice noodle digesta with higher amylose contents exhibited greater viscosities throughout the simulated oral-gastric-intestinal digestion steps. The flow behaviors of the rice noodle digesta consisted of the Power-law region and infinite shear plateau that were satisfactorily characterized by the Sisko model (R² > 0.99).