添加芽孢杆菌对草鱼池塘中真核微生物的影响

为了研究添加芽孢杆菌对池塘中真核微生物群落结构和理化因子的影响,采用高通量测序技术分析了实验组(添加芽孢杆菌池塘)与对照组(普通池塘)水体真核微生物群落结构,同时分析了两组池塘的水体理化指标。结果表明:8、9月实验组池塘水体中TN、NH_4~+-N、NO_3~--N含量显著低于对照组(P0.05)。水体中梅尼小环藻(Cyclotella meneghiniana)、红囊藻(Hedriocystis)、蓝隐藻(Chroomonas)、丝孢酵母属(Trichosporon)和小环藻属(Cyclotella)真核微生物丰度显著高于对照组(P0.05)。实验组池塘水体真核微生物Chao1指数和Shannon指数显著高于对照组(P0.05)。实验结果证实:通过向池塘添加芽孢杆菌,可以改变水体中真核微生物群落的结构,从而实现对池塘理化因子的调节。研究结果对于降低水产养殖尾水对水域环境的污染具有一定的意义。     

Long-term fertilization leads to specific PLFA finger-prints in Chinese Hapludults soil

Soil microbes play essential roles in the biogeochemical processes of organic carbon and nutrient cycling. Many studies have reported various short-term effects of fertilization on soil microbes. However, less is known about the effects of longterm fertilization regimes on the rhizosphere. Therefore, the objective of this study was to explore how the soil microbial communities in the rhizosphere respond to different long-term fertilization strategies. Based on a 21-year field treatment experiment in Guizhou, China, we extracted phospholipid fatty acids(PLFAs) to determine the microbial community structure in both the non-rhizosphere(NR) and rhizosphere(R). Six treatments were included: no fertilizer(CK), mineral nitrogen fertilizer(N), N with potassium(NK), phosphorus with K(PK), NPK, and NPK combined with manure(MNPK). The results showed that total PLFAs under unbalanced mineral fertilization(N, NK and PK) were decreased by 45% on average in the NR compared with CK, whereas MNPK increased fungi and G~– bacteria abundance significantly in both the NR(by 33 and 23%) and R(by 15 and 20%), respectively. In addition, all microbial groups in the R under these treatments(N, NK and PK) were significantly increased relative to those in the NR, except for the ratio of F/B and G~+/G~–, which might be due to the high nutrient availability in the R. Soil pH and SOC significantly regulated the soil microbial community and structure, explaining 51 and 20% of the variation in the NR, respectively. However, the rhizosphere microbial community structure was only significantly affected by soil pH(31%). We concluded that the soil microbial community in the NR was more strongly affected by long-term fertilization than that in the R due to the rhizosphere effect in the agricultural ecosystem. Rhizosphere nutrient conditions and buffering capacity could help microbial communities resist the change from the long-term fertilization.     

Community structure and carbon and nitrogen storage of sagebrush desert under grazing exclusion in Northwest China

Overgrazing is regarded as one of the key factors of vegetation and soil degradation in the arid and semi-arid regions of Northwest China.Grazing exclusion(GE)is one of the most common pathways used to restore degraded grasslands and to improve their ecosystem services.Nevertheless,there are still significant controversies concerning GE’s effects on grassland diversity as well as carbon(C)and nitrogen(N)storage.It remains poorly understood in the arid desert regions,whilst being essential for the sustainable use of grassland resources.To assess the effects of GE on community characteristics and C and N storage of desert plant community in the arid desert regions,we investigated the community structure and plant biomass,as well as C and N storage of plants and soil(0-100 cm depth)in short-term GE(three years)plots and adjacent long-term freely grazing(FG)plots in the areas of sagebrush desert in Northwest China,which are important both for spring-autumn seasonal pasture and for ecological conservation.Our findings indicated that GE was beneficial to the average height,coverage and aboveground biomass(including stems,leaves and inflorescences,and litter)of desert plant community,to the species richness and importance values of subshrubs and perennial herbs,and to the biomass C and N storage of aboveground parts(P<0.05).However,GE was not beneficial to the importance values of annual herbs,root/shoot ratio and total N concentration in the 0-5 and 5-10 cm soil layers(P<0.05).Additionally,the plant density,belowground biomass,and soil organic C concentration and C storage in the 0-100 cm soil layer could not be significantly changed by short-term GE(three years).The results suggest that,although GE was not beneficial for C sequestration in the sagebrush desert ecosystem,it is an effective strategy for improving productivity,diversity,and C and N storage of plants.As a result,GE can be used to rehabilitate degraded grasslands in the arid desert regions of Northwest China.     

长期少免耕对中国东北玉米农田土壤呼吸及碳氮变化的影响

在黑土区37年不同耕作模式定位试验田,采用原位法比较常规旋耕灭茬起垄（CT）、旋耕留高茬行间深松-少耕（RT）、免耕（NT）和深翻（PT）4种耕作模式土壤呼吸速率及其与土壤温度和湿度的关系,测定土壤总碳氮、无机氮,微生物碳氮的变化。结果表明,玉米生育期间土壤呼吸呈单峰变化,开花期达最大值,生长季平均呼吸速率依次为RT > NT > CT > PT。4个处理土壤温度与土壤呼吸速率间存在显著指数关系。不同耕作模式0~10 cm土壤温度可解释土壤呼吸速率变异的38.3%~67.9%,温度敏感性系数Q10范围为2.1~5.3;RT和NT处理显著提高0~20 cm土壤碳氮含量;RT处理在0~20 cm土壤中微生物碳含量均高于CT、NT和PT处理。RT处理土壤呼吸对温度响应提高,RT和NT处理显著增加上层土壤总碳氮含量,利于土壤质量提升。     

Mouse melanoma cell exosomes promote expression of Rac1 protein in fibroblasts

AIM To investigate the effect of exosomes secreted by mouse melanoma cells on the expression of Ras-related C3 botulinum toxin substrate 1 (Rac1) protein in fibroblasts. METHODS Ultracentrifugation was adopted to separete exosomes secreted by mouse melanoma B16-F10 cells. The morphological structure of exosomes was observed by negative-staining electron microscopy. The size distribution of exosomes was determined by nanoparticle tracking analysis (NTA). The exosomal markers, tumor susceptibility gene 101 (Tsg101) and tyrosinase-related protein 2 (Tyrp2), were identified by Western blot. Laser confocal microscopy was used to observe the process that mouse embryonic fibroblasts (MEF) took in exosomes during co-culture. Immunocytochemical staining and Western blot were used to detect the expression of Rac1 protein in MEF. RESULTS B16-F10 cell exosomes showed a typical tea tray-like structure, with a size range of 141~255 nm, and expressed protein markers Tsg101 and Tyrp2. The results of laser confocal microscopy showed that compared with co-culture at 0 h, a small number of exosomes appeared in the MEF at 12 h, and a large number of exosomes accumulated in the MEF after co-cultured for 24 and 36 h. Western blot analysis showed that compared with co-culture at 0 h, the expression of Rac1 protein in the MEF was significantly increased at 24 h and 36 h of co-culture (P<0.01). The results of immunocytochemical staining showed that compared with co-culture at 0 h, the positive expression level of Rac1 in the MEF cells was significantly increased at 12 h, 24 h and 36 h of co-culture (P<0.05 or P<0.01). CONCLUSION Intake of exosomes secreted by mouse melanoma cells promotes the expression of Rac1 protein in fibroblasts.     

民勤4种主要灌木植物δ13C季节变化及与气象因子的关系

对民勤4种主要灌木植物叶片/同化枝δ¹³C测定,研究不同荒漠灌木植物叶片/同化枝δ¹³C差异及其季节变化,并探讨植物叶片/同化枝δ¹³C与气象因子之间的关系。结果表明:1）4种灌木植物叶片/同化枝δ¹³C值不同月份之间存在极显著差异（P＜0.01）,除梭梭外,其他3种植物δ¹³C值生长初期（5月）>生长中后期（6-10月）。4 种灌木植物叶片/同化枝δ¹³C值种间差异在5-10月都达到极显著水平（P＜0.01）,沙拐枣＞梭梭＞柠条锦鸡儿＞白刺。2）4种主要灌木植物叶片/同化枝δ¹³C值受到物种、月份、物种×月份的影响,均达到极显著水平(P＜0.01),但受物种的影响最大。3）4种灌木植物叶片/同化枝δ¹³C值与气象因子的关系因其物种而异;影响梭梭、沙拐枣、白刺、柠条锦鸡儿叶片/同化枝δ¹³C值的主要气象因子分别是平均气温、相对湿度、蒸发量。     

不同增氧滴灌方式对蔬菜生长生理指标的影响

为了研究不同增氧方式对盆栽小白菜生长生理指标的影响,以小白菜为供试作物,采用盆栽地下滴灌的方式,以普通地下滴灌作为对照(CK),设置循环曝气(MAI)、双氧水(H₂O₂)、纯氧扩散器曝气(OC)及射流振荡器曝气(FO)4个增氧灌溉处理.结果表明,增氧地下滴灌显著提高了土壤呼吸速率,处理MAI,OC和FO较对照处理分别增大了65.87%,66.79%和111.62%.增氧地下滴灌促进了小白菜的根系生长、光合作用、蒸腾速率和气孔导度,进而提高了小白菜的物质量积累和产量.与对照相比,处理MAI的地下部鲜质量增大了42.03%,地下部干质量增大了79.85%;处理MAI,H₂O₂,OC和FO的光合速率分别增大了868.62%,794.14%,778.67%和650.19%;处理MAI,H₂O₂和OC的气孔导度较CK增大了157.14%,128.57%和85.71%,蒸腾速率增大了55.61%,32.38%和19.58%;处理MAI和H₂O₂的产量分别增大了56.36%和38.72%.综上,增氧地下滴灌可增强小白菜根区的土壤呼吸作用,改善光合作用、蒸腾速率和气孔导度,提高了产量及水分利用效率.其中,循环曝气处理的改善效果最为显著.     

生物炭连续施用对农田土壤氮转化微生物及N2O排放的影响

【目的】研究连续添加生物炭6年后对农田土壤氮转化相关微生物功能基因的影响,揭示生物炭影响作物产量和N₂O排放的微生物学机制,并为生物炭的推广使用提供理论依据。【方法】通过在潮土农田设置0（BC0,对照)、2.25（BCL,低量）、6.75（BCM,中量）和11.25 t·hm^-2（BCH,高量）4个秸秆生物炭量处理的田间定位试验,采用田间观测、化学分析、荧光定量PCR（qPCR）技术,系统研究施用生物炭对氧化亚氮（N₂O）排放、氨单加氧酶（amoA）、亚硝酸还原酶（nirK、nirS）、氧化亚氮还原酶（nosZ）基因丰度及夏玉米产量的影响。【结果】与对照BC0处理相比,施用生物炭可显著提高夏玉米籽粒产量,且BCM处理籽粒产量达到最大值10 811 kg·hm^-2,显著降低夏玉米生育期N₂O累积排放量,并以BCM处理减少N₂O排放效果最优。研究还发现,在夏玉米多个生育时期,与对照比较,生物炭施用可以显著提高耕层土壤无机氮储量和土壤含水量。此外,随着生物炭施用量增加,土壤氨氧化古菌（AOA）基因拷贝数在夏玉米大喇叭口期和成熟期均表现为先上升后下降趋势,且两个时期均以BCM处理最高,而氨氧化细菌（AOB）基因拷贝数在夏玉米大喇叭口期和成熟期分别为BCH处理和BCM处理最高。与对照相比,中、高量生物炭施用（BCM、BCH处理）可显著提高夏玉米大喇叭口期和成熟期土壤反硝化作用功能相关基因（nirK、nirS、nosZ）拷贝数。相关性分析表明,夏玉米成熟期土壤N₂O排放通量与土壤硝态氮、土壤含水量、AOA、AOB、nirK、nirS、nosZ呈显著负相关关系。【结论】施用生物炭通过增加土壤微生物氮转化功能基因丰度进而降低土壤N₂O排放,通过增加土壤耕层无机氮储量和土壤水分含量进而提高作物产量,并以中等用量（6.75 t·hm^-2）施用效果最优。     

鸡粪中雌激素的检测及微生物降解

为检测鸡饲料和鸡粪中雌激素,优化其微生物降解条件,应用高效液相色谱（HPLC）法检测蛋鸡饲料和鸡粪中的雌激素,利用11种高效降解雌激素的单一菌株和等体积混合得到的菌群开展雌激素的降解实验。结果表明：只有蛋鸡鸡粪中含有雌二醇（E2）,含量为230 ng·g^-1。微生物菌群的降解率高于单一菌株,在27℃下,10 d内能够降解76%的E2。研究表明,蛋鸡鸡粪中有雌激素存在,微生物菌群对雌激素的降解率高于单一菌株。