Dynamics of Cotton Rhizosphere Bacterial Community Structure in Cotton Continuous Cropping Field Soil

[Objective] The aim of this study is to understand the dynamics of cotton rhizosphere bacterial community structure in cotton continuous cropping field soil. [Method] 16S rDNA genes were sequenced by high-throughput sequencing technology to determine the community structure of cotton rhizosphere bacteria in different developmental stages using an upland cotton cultivar (Gossypium hirsutum cv. TM-1). [Result] Four dominant phyla were found in the cotton rhizosphere bacterial community including Proteobacteria, Acidobacteria, Planctomycetes, and Bacteroidetes. The four dominant phyla and Firmicutes were largely influenced by cotton root. The relative abundances of Acidobacteria, Planctomycetes, Bacteroidetes were promoted, and Proteobacteria and Firmicutes were inhibited, by cotton root. There were significant differences in community structure, but not species richness or α-diversity among different developmental stages of the cotton rhizosphere bacterial community; the differences between the flowering stage and the budding stage were greater than the differences between the budding stage and the seedling stage. The diversity of the rhizosphere bacterial community in cotton continuous cropping field soil was significantly higher than that of the bulk soil; the β-diversity values of both the rhizosphere and bulk soil bacterial communities were highest in the flowering stage. [Conclusion] The structure and dynamics of the cotton rhizosphere bacterial community in cotton continuous cropping field soil was defined by high-throughput sequencing. The effect of cotton on the rhizosphere bacterial community structure was most significant in the flowering stage.     

金龟子绿僵菌在草原羊草和克氏针茅根际的种群动态和根内宿存鉴定

绿僵菌防治草原蝗虫效果显著,其在草原田间的存活能力影响其持续控害效果。除了侵染昆虫,绿僵菌还具有在植物根际宿存和根内生的生活方式,但相关宿存规律和内生性的研究报道很少。本文研究了绿僵菌在内蒙古草原两种优势种植物羊草和克氏针茅根际的种群数量变化并对其在这两种草的根内宿存进行了鉴定。结果表明,在干旱的内蒙古草原,绿僵菌施用后种群数量在30 d内快速下降,但能够以低密度在羊草、克氏针茅根际土壤中至少延续宿存75 d,羊草根际环境比较利于绿僵菌生存。对菌株egfp基因标记的特异PCR检测证明了绿僵菌在羊草和克氏针茅根内宿存。试验数据为指导植保生物防治中充分利用绿僵菌的昆虫病原性和植物内生特性提供理论基础,也将成为绿僵菌物种生活方式多样性、与植物互作及共进化研究的重要基础。     

高粱绿肥种植密度对设施黄瓜根系生长相关因子的影响

以设施黄瓜为研究对象,在黄瓜定植前40 d分别播撒240、120、60 kg/hm~2不同密度高粱种子,并于黄瓜定植前将高粱植株作为绿肥翻入土壤,研究其对黄瓜结果期生长指标、根际土壤养分含量以及细菌群落结构变化的影响,探讨其影响黄瓜生长指标、根际养分含量和菌群结构变化的内在机制。试验结果表明:240 kg/hm~2种植密度的高梁绿肥可显著提高黄瓜根际土壤有机质质量比31.66%、碱解氮质量比8.75%、速效磷质量比13.89%、速效钾质量比16.86%、脲酶活性6.85%,显著提高结果期黄瓜茎粗12.83%、根系数15.81%、根总体积24.11%、叶片总面积11.43%、单株结瓜数10.97%、总产量9.81%;240 kg/hm~2种植密度的高粱绿肥可明显提高黄瓜根际土壤中变形菌门、放线菌门、厚壁菌门比例而降低酸杆菌门、绿弯菌门、拟杆菌门比例,明显提高鞘脂单胞菌科、黄色单胞菌科、红螺菌科、中华杆菌科、生丝微菌科比例而降低Gp6科、Gp16科、Gp4科比例。该处理主要通过降低根际土壤绿弯菌门和提高鞘脂单胞菌科、中华杆菌科比例而提高脲酶活性,通过提高厚壁菌门、黄色单胞菌科、红螺菌科和降低根际土壤酸杆菌门比例而促进碱解氮质量比、速效磷质量比、茎粗、叶片总面积、根系数、根总体积的提高,最终促进黄瓜生长并提高产量。     

连栽木麻黄根际微生物群落结构和功能特征

以不同栽培代数的木麻黄(第1代FCP、第2代SCP、第3代TCP)根际土壤为试验材料,运用BIOLOG微平板和磷脂脂肪酸(PLFA)技术分析根际土壤微生物群落结构和功能多样性对多代连栽响应。结果表明,不同代数的土壤微生物对各类碳源的利用程度存在显著差异,连栽后木麻黄根际土壤微生物对碳源利用率显著下降。在6类碳源中,除胺类外,其他5种碳源均呈现FCP>SCP>TCP。PLFA分析共检测到11种PLFA生物标记,FCP土壤微生物PLFA生物标记总量明显高于SCP和TCP,3个代数土壤中含量最高的PLFA生物标记是i16:0、a15:0和18:1ω9c。土壤中特征微生物含量差异明显,细菌分布量最大,其次是真菌和放线菌。随栽植代数增加,细菌含量减少,真菌含量增加。土壤微生物群落多样性指数均呈现FCP>TCP>SCP,与土壤理化性质变化密切相关。可见木麻黄连栽显著影响其根际土壤微生物群落结构与功能,因此根际土壤微生态失衡可能是导致木麻黄连栽障碍的重要因素。     

Soil Characteristics Overwhelm Cultivar Effects on the Structure and Assembly of Root-Associated Microbiomes of Modern Maize

Modern breeding primarily targets crop yield traits and is likely to influence root-associated microbiomes, which play significant roles in plant growth and health. The relative importance of soil and cultivar factors in shaping root-associated microbiomes of modern maize (Zea mays L.) remains uncertain. We conducted a pot experiment in a controlled environment using three soils (Mollisol, Inceptisol, and Ultisol) and four contrasting cultivars, Denghai 605, Nonghua 816, Qiaoyu 8, and Zhengdan 958, which are widely planted in China. We used 16S rRNA gene amplicon sequencing to characterize the bacterial communities in the bulk soil, rhizosphere, and endosphere. Our results showed that the four cultivars had different shoot biomass and root exudate total organic carbon and organic acid contents. The microbiomes in the bulk soil, rhizosphere, and endosphere were different. We observed apparent community divergence between soils rather than cultivars, within which edaphic factors substantially contributed to microbiome variation. Moreover, permutational multivariate analysis of variance corroborated significant contributions of soil type but not cultivar on the root-associated microbiome structure. Differential abundance analysis confirmed that each soil presented a distinct root microbiome, while network analysis indicated different co-occurrence patterns of the root microbiome among the three soils. The core root microbiome members are implicated in plant growth promotion and nutrient acquisition in the roots. In conclusion, root-associated microbiomes of modern maize are much more controlled by soil characteristics than by cultivar root exudation. Our study is anticipated to help improve breeding strategies through integrative interactions of soils, cultivars, and their associated microbiomes.     

转cry1Ie基因抗虫玉米对根际微生物群落碳代谢的影响

转基因作物对土壤生态系统的影响日益引起人们的广泛关注。采用Biolog ECO微平板对转基因抗虫玉米IE09S034及其对应的受体玉米根际微生物群落碳代谢功能进行了比较研究。结果表明,转基因抗虫玉米IE09S034对根际微生物碳代谢功能无显著性影响,且发现年份和生育期是引起根际微生物碳代谢情况变化的主要因素。本文为转基因抗虫玉米IE09S034的环境安全性评估提供了新的依据。     

薄膜扩散梯度（DGT）技术在环境微界面物质运移过程研究中的应用

土壤、沉积物、水体和生物体之间的接触和作用形成了多种环境微界面。这些环境微界面是物质迁移转化的重要场所,而高度时空异质性的界面特征使得对其中化学反应信息的捕捉变得极其复杂且困难。薄膜梯度扩散(DGT)技术以其原位测量元素生物有效态和高空间分辨率等优势,适用于研究化学异质性的界面过程。本文系统总结了DGT技术在环境微界面的物质运移过程研究中的应用现状,包括以下3方面内容:一是一维物质浓度测定;二是二维化学分布成像;三是与薄膜扩散平衡技术(DET)、平衡式孔隙水采样器(Peeper)和平面光极(PO)等技术联用同步获取多种溶质分布信息。现有研究证据表明,DGT适合在亚毫米(几十至几百微米)至毫米尺度研究环境微界面营养盐和污染物运移的生物地球化学过程,并可与其他化学成像技术结合研究物质跨界面运移的驱动因子和动力学特征。最后,在DGT技术发展与应用场景扩展等方面提出了几点展望。     

辣椒种植对根际土壤微生物多样性的影响

利用Illumina Mi Seq 2500测序平台对2个辣椒自交系(SJ11–3、06g19–1–1–1)和1个地方品种(樟树港)种植前后根际土壤的微生物进行测序,并结合相关生物信息学分析土壤细菌16S r RNA基因V3+V4区域和真菌ITS1区域的丰富度、多样性指数以及微生物群落结构。结果表明:辣椒种植前后土壤均以变形菌门和酸杆菌门为优势细菌,子囊菌门、担子菌门和接合菌门为优势真菌;辣椒种植前后土壤中的细菌种类较真菌更加丰富,多样化程度更高;种植辣椒后土壤细菌、真菌的相对丰度及群落结构都发生了变化,土壤细菌Ace、Chao和Shannon指数都有所增加,增幅分别为25.26%~25.49%、24.51%~24.87%和8.28%~9.55%,而真菌的Shannon指数有不同程度降低,Simpson指数增大,增幅78.45%~275.69%;辣椒种植后土壤中细菌和真菌的操作分类单元(OTU)数和类型也有所改变,细菌OTU个数较种植前明显升高,增幅约28.73%,土壤中特有的OTU类型明显减少;真菌OTU个数变化及类型变化比细菌大,病原性真菌的相对丰度大幅增加。     

棘孢木霉菌肥对黄瓜枯萎病的防治效果及对连作黄瓜根际土壤微生物种群的影响

为明确棘孢木霉Trichoderma asperellum菌肥在防治黄瓜枯萎病的同时对连作黄瓜根际土壤微生物种群的影响,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)和磷脂脂肪酸(phospholipid fatty acid,PLFA)分析方法分别测定了棘孢木霉菌肥对连作4年的黄瓜根际棘孢木霉菌和尖孢镰孢菌Fusarium oxysporum DNA拷贝数变化和黄瓜根际微生物种群的影响。结果表明:棘孢木霉菌肥对黄瓜幼苗期和速长期黄瓜枯萎病的防治效果分别为70.24%和76.81%,均与药剂对照的防效差异不显著。棘孢木霉菌的DNA拷贝数出现2个高峰期,即黄瓜苗期和盛果期,其DNA拷贝数分别为235 000.00 ng/μL和80 500.00 ng/μL。黄瓜速长期、盛果期、生长末期的尖孢镰孢菌DNA拷贝数分别为15.41、54.87和18.36 ng/μL,且显著低于同一时期药剂对照和清水对照。黄瓜幼苗期、速长期、开花期和盛果期根际土壤微生物丰度分别为2.24、1.98、2.52和2.12,均高于药剂对照。由此可见,棘孢木霉菌肥在防治黄瓜枯萎病的同时,还可以改善黄瓜连作土壤微生物种群结构。     

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.