水稻根际固氮菌促生特性评价

利用半固体培养基和土壤栽培经水稻特异性根际固氮菌处理的水稻幼苗,发现固氮菌属(Azotobacterspp.)菌株R-N00、R-N1110和R-N1112具有较强的促生能力。半固体培养基栽培获得接种3株菌株的水稻幼苗干重分别提高16.60%、9.23%和9.37%,根干重分别提高38.00%、30.00%和55.00%;土壤栽培获得接种菌株的水稻幼苗干重分别提高93.25%、90.18%和97.21%,根干重分别提高71.25%、72.21%和74.92%。这3株菌的固氮能力分别为13.37、11.93和13.57 mg.L-1;合成铁载体能力均达到5+;分泌吲哚乙酸分别为6.79、8.60和3.39 mg.L-1;溶磷能力分别为41.63、37.76和39.78 mg.L-1。结果表明,水稻根际促生菌的各种促生生理活性共同作用促进水稻幼苗生长,为高效生物肥料的研究提供参考。     

新造地马铃薯根际固氮解磷微生物的分离与鉴定

【目的】揭示马铃薯根际土壤固氮解磷菌类群,分析其固氮酶活性和溶磷能力,为利用新造地作物根际促生菌提供依据。【方法】以采自延安市安塞区新造地的马铃薯根际土壤为供试样品,采用Ashby培养基和无机磷培养基分离纯化马铃薯根际固氮解磷菌,以16S rDNA基因分析马铃薯根际微生物类群组成,以乙炔还原方法测定其固氮酶活性,以溶菌圈法和钼蓝比色法测定其溶磷能力。【结果】从延安新造地马铃薯根际土壤中分离并筛选到9株固氮解磷菌,经鉴定分别属于假单胞杆菌属(Pseudomonas)、克雷伯氏菌属(Klebsiella)、红球菌属(Rhodococcus)、节杆菌属(Paenarthrobacter)、芽孢杆菌属(Bacillus)、贪噬菌属(Variovorax)、肠杆菌属(Kosakonia)和副球菌属(Paracoccus)等8个属;9株固氮解磷菌均具有固氮和溶磷特性,其固氮酶活性在11.88～95.08 nmol/h,其中菌株N34的固氮酶活性最高,且与其他菌株的固氮酶活性差异显著;9株固氮解磷菌株溶磷能力存在一定的差异,溶磷能力为22.05～54.9 mg/L,其中菌株N46的溶磷能力最好,7 d累积溶磷能力达到54.9 mg/L。【结论】分离纯化得到的9株固氮解磷菌均具有一定的固氮解磷能力,可用于开发马铃薯功能菌肥。     

植物根际促生菌的研究进展及其环境作用

具有固氮、溶磷、产生植物激素和分泌抗生素等功能的植物根际促生菌已成为新型生物肥料开发的研究热点.介绍了植物根际促生菌的定义、主要种类、作用机制、研究和应用现状.并提出绿色食品生产、资源再生利用要与植物根际促生菌生产相结合,促进绿色农业的发展,表明促生菌在经济与环境协调发展中具有重要的理论和现实意义.     

联合固氮酶活性测定中酶促反应时间的控制方法研究

乙炔还原法是国际上普遍公认的测定生物固氮活性的方法,该法具有简便、快速、灵敏度高等优点.乙炔还原反应的温育时间不同,直接影响乙炔还原法测定固氮酶活性结果的可比性.为确保大批量的联合固氮酶活性测定条件的一致性,有必要选择一种固氮酶活性阻断剂,使联合固氮体系中的乙炔还原反应维持在特定时间后予以阻断.本文比较了3种灭菌剂对2株联合固氮菌的抑菌效果及其对水稻根际联合固氮酶活性阻断效果,结果表明,3.5%甲醛水溶液是一种理想的联合固氮酶活性阻断剂.     

水稻根际固氮菌NX-2的分离鉴定及其培养条件优化

筛选高效固氮能力的植物根际促生菌是研发微生物肥料的关键。研究从水稻根际土壤中分离筛选得到一株固氮菌NX-2,利用乙炔还原法测定其固氮酶活可达16.18μmol/(m L·h)(以C2H4浓度计)。通过形态观察、生理生化特性和16S r DNA序列分析,菌株NX-2被鉴定为褐球固氮菌。进一步研究其生长特性,结果显示最佳培养条件为培养温度28℃、初始p H值6.5、接种量3%。     

联合固氮工程菌E7在蔬菜上的应用效果

联合固氮工程菌E7是定植在植物根际的固氮细菌,其主要作用是能产生一些生理活性物质刺激植物发根和生长,增强植物对营养物质的吸收能力,同时固定空气中的氮气为有机氮,菌体死亡后其体内的有机氮可被植物吸收利用,增加植物的氮素营养.由于施用固氮菌后可节省部分氮肥,从而减轻了由于氮肥的过多施用而造成的环境污染.联合固氮工程菌E7在玉米、水稻等作物多年试验示范中效果显著,为研究联合固氮工程菌E7在蔬菜上的应用效果,2001年进行了本试验.     

根际联合固氮菌对小麦生产的效应研究

在盆栽条件下,施用专性小麦根际联合固氮菌,提高了根际土固氮酶活性,植株叶绿素和全氮含量以及小玫生物产量,增加了小麦根际土和根表皮层中联合固氮菌数量;净固氮量比下不施菌增加４５－６０ｋｇ／ｈｍ＾２,与固氮效力较高的巴西固氮螺菌相当,但适应性较优。     

几种木本植物联合固氮细菌的研究—联合固氮细菌的分离鉴定及固氮活性

在几种木本植物杨树，杉木，油茶根际存在着联合固氮体系，从它们根际分离到26株具有较高固氮酶活性的菌株，筛选了4株具有高固氮活性的菌株Aa33,Ba41,y12,Y22进行固氮特性研究，并选择菌株y12,Y22进行形态，培养特征及生理生化研究，初步鉴定为黄杆菌属Flavobacterium和产碱菌属Alcaligenes，这对今后开展木本植物固氮领域的研究及其利用具有现实的指导意义。     

盐碱胁迫下水稻促生菌的筛选

水稻在生长过程中常受到各类非生物胁迫的影响,例如土壤盐碱化、干旱等,进而导致水稻大量减产。为促进盐碱胁迫下植物的生长,从黑龙江省安达盐碱地植物根际土壤中分离得到了6株植物根际促生菌（PGPR）,并对其促生作用进行初步研究。结果表明,分离到的6株植物根际促生菌能够显著促进拟南芥、水稻和玉米在盐碱条件下的生长,使生物量大幅提高。这些PGPR均属于芽孢杆菌属（Bacillus）,有良好的抗逆性,其分泌的促生物质不尽相同,说明这些根际促生菌具有不同的促生机制。     

几种菌株对水稻的促生能力测定

水稻在我国粮食生产中占有重要的地位,植物根际促生菌(PGPR)是促进植物生长和提高土壤肥力的关键因素。采用盆栽育苗试验、MS半固体培养基育苗试验以及田间试验,验证东北寒地水稻根际促生菌对水稻生长的促生效果。应用9株水稻根际促生菌,研究其对水稻生长的影响,9株根际促生菌可以在土壤中促进水稻植株的生长,其中Bacillus aryabhattai(LZP01)、Bacillus pumilus(LZP02)、Bacillus megaterium(LZP03)促生效果最好,在水稻株高、茎粗、地上质量和根表面积等指标上均有显著促生效果。MS半固体培养基育苗试验结果可以发现,3株优势根际促生菌对水稻生长指标和根系指标具有促生作用。于黑龙江省二九零农场进行LZP01、LZP02、LZP03复合菌株的田间试验,加入复合菌株的土壤生长出来的水稻与对照相比盘根能力较好,更加适合田间环境生长。结果将对水稻微生物肥料开发利用产生推动作用,在农业生产实践中产生经济效益、社会效益和生态效益。     

酸性土壤上花生高效根瘤菌的分离及应用

【背景】 花生（Arachis hypogaea L.）是世界上重要的油料和经济作物。在我国南方产区,大部分花生种植地土壤为酸性。酸性土壤不仅pH值低、瘦瘠,而且还有低磷、铝毒等诸多障碍因素,严重限制了花生的生物固氮及产量。【目的】 本文旨在分离及应用适应酸性土壤的高效固氮根瘤菌,提高花生固氮效率及产量,改良酸性土壤。【方法】 利用平板划线结合镜检的方法,从田间采集的新鲜花生根瘤中分离纯化单菌落;通过PCR技术检测分离物中是否含有结瘤基因nodA和固氮基因nifH,进行根瘤菌的初步分子鉴定;再通过16S rRNA基因序列比对,对根瘤菌进行进一步分子鉴定。候选根瘤菌通过水培回接,检测其与花生的共生结瘤及固氮能力;再通过田间试验评价筛选出来的候选根瘤菌在酸性土壤上的应用效果。【结果】 本研究首先从不同酸性土壤种植区域的花生根瘤中分离、纯化得到256个分离物;其中10株含有nodA和nifH,初步确定为根瘤菌。经16S rRNA基因全长序列测定,发现8株为慢生型根瘤菌（Bradyrhizobium）,2株为根瘤菌属根瘤菌（Rhizobium）。水培回接试验发现,这10株根瘤菌均能够与花生共生、形成有效根瘤,证实是花生根瘤菌。在此基础上,选取4株固氮效率较高的根瘤菌,在酸性土壤上应用。结果表明,4株根瘤菌均能与花生在酸性土壤上形成根瘤,而未接种的花生根部不能形成根瘤。并且接种根瘤菌后显著改善了花生氮营养,提高了花生的生物量和产量。与不接种的对照相比,接种根瘤菌后,花生生物量、产量和氮含量分别提高了27.1%—38.0%、24.7%—104.2%和73.9%—151.3%。【结论】 本研究分离鉴定的花生根瘤菌能够高效固氮和适应酸性土壤,具有重要的应用前景。     

Transfer characteristics of nitrogen fixed by leguminous green manure crops when intercropped with maize in northwestern China

To ascertain the possibility of cultivating maize using biological nitrogen fixation (BNF) by leguminous green manure crops in maize/leguminous green manure intercropping systems, BNF and nitrogen (N) transfer were studied in Xining and Wuwei, two typical northwestern Chinese cities. The experimental treatments included monocultured maize, monocultured green manures (hairy vetch and common vetch), and their intercropping systems. The proportions of N derived from the atmosphere (%N_dfa) in intercropping systems were not significantly different from that in monocultured green manure systems at either experimental site, except for that in hairy vetch (HV) in Xining. The amount of N derived from the atmosphere (N_dfa) of common vetch (CV) significantly decreased from 1.16 and 1.10 g/pot in monoculture to 0.77 and 0.55 g/pot when intercropped with maize, in Xining and Wuwei, respectively, and the N_dfa of HV when intercropped significantly decreased from 1.02 to 0.48 g/pot in Xining. In the intercropping systems in Xining and Wuwei, the amounts of N transferred (N_transfer) from CV to maize were 21.54 and 26.81 mg/pot, accounting for 32.9 and 5.9% respectively of the N accumulation in maize, and the values of N_transfer from HV to maize were 39.61 and 46.22 mg/pot, accounting for 37.0 and 23.3%, respectively, of the N accumulation in maize. Path analysis showed that soil nutrient and green manure biomass were mainly related to N_dfa, and that δ¹⁵N had a primary relationship with N_transfer. We found that 5.9–37.0% of N accumulation in maize was transferred from green manures, and that the N transfer ability to maize of HV was higher than that of CV. In conclusion, intercropping with leguminous green manures provided a feasible way for maize to effectively utilize biologically fixed N.     

接种根瘤菌对豌豆/玉米体系根际细菌数量及氮营养的影响

采用盆栽试验研究了单作及复合体系中接种根瘤菌对土壤含氮量,植株氮素积累量及豌豆、玉米根际细菌数量的影响.结果表明:接种根瘤菌显著增加了土壤含氮量和植株氮素积累量,当地筛选菌株XC3.1的接种效果优于菌株ACCC16l01,与未接种根瘤菌和接种ACC16101相比,接种菌株XC3.1的土壤含氮量和植株氮素积累量分别增加2...     

Nitrogen acquisition,fixation and transfer in maize/alfalfa intercrops are increased through root contact and morphological responses to interspecies competition

Nitrogen(N) fixation by legumes and nitrogen transfer to cereals have been considered as important pathways for overyielding and higher N use efficiency in cereal/legume intercropping systems. However, the extent to which root morphology contributes to N fixation and transfer is unclear. A two-factorial greenhouse experiment was conducted to quantify the N fixation, transfer and root morphology characteristics of the maize/alfalfa intercropping system in two consecutive years using the 15 N-urea leaf labeling method, and combining two N levels with three root separation techniques. N application could inhibit N fixation and transfer in a maize/alfalfa intercropping system. Irrespective of the N application level, compared with plastic sheet separation(PSS), no separation(NS) and nylon mesh separation(NNS) significantly increased the total biomass(36%) and total N content(28%), while the N fixation rate also sharply increased by 75 to 134%, and the amount of N transferred with no root barrier was 1.24–1.42 times greater than that with a mesh barrier. Redundancy analysis(RDA) showed that the crown root dry weight(CRDW) of maize and lateral root number(LRN) of alfalfa showed the strongest associations with N fixation and transfer. Our results highlight the importance of root contact for the enhancement of N fixation and transfer via changes in root morphology in maize/alfalfa intercropping systems, and the overyielding system was achieved via increases in maize growth, at the cost of smaller decreases in alfalfa biomass production.     

禾谷类作物固氮研究新进展

对禾谷类作物自身固氮、共生固氮和联合固氮作用等方面的研究工作进行了评述,并提出应该注意的问题及可能解决的途径。     

湖北省花生平衡施肥技术研究Ⅵ.花生氮肥用量

在田间条件下研究了不同氮肥用量对花生产量和固氮的影响。结果表明,在红安低氮土壤上,施氮使花生产量和氮积累量显著增加,但固氮比例快速降低;在蔡甸高氮土壤上,施氮对花生产量和氮积累量影响不明显,固氮比例变化不大。建议生产上应根据土壤条件确定施氮策略,即低氮土壤上优先考虑增加花生产量来施氮,在高氮土壤上优先考虑发挥花生固氮能力来施氮。     

Effect of nitrogen fertilization on yield,N content,and nitrogen fixation of alfalfa and smooth bromegrass grown alone or in mixture in greenhouse pots

Planting grass and legume mixtures on improved grasslands has the potential advantage of realizing both higher yields and lower environmental pollution by optimizing the balance between applied N fertilizer and the natural process of legume biological nitrogen fixation. However, the optimal level of N fertilization for grass-legume mixtures, to obtain the highest yield, quality, and contribution of N2 fixation, varies with species. A greenhouse pot experiment was conducted to study the temporal dynamics of N2 fixation of alfalfa(Medicago sativa L.) grown alone and in mixture with smooth bromegrass(Bromus inermis Leyss.) in response to the addition of fertilizer N. Three levels of N(0, 75, and 150 kg ha–1) were examined using 15N-labeled urea to evaluate N2 fixation via the 15 N isotope dilution method. Treatments were designated N0(0.001 g per pot), N75(1.07 g per pot) and N150(2.14 g per pot). Alfalfa grown alone did not benefit from the addition of fertilizer N; dry matter was not significantly increased. In contrast, dry weight and N content of smooth bromegrass grown alone was increased significantly by N application. When grown as a mixture, smooth bromegrass biomass was increased significantly by N application, resulted in a decrease in alfalfa biomass. In addition, individual alfalfa plant dry weight(shoots+roots) was significantly lower in the mixture than when grown alone at all N levels. Smooth bromegrass shoot and root dry weight were significantly higher when grown with alfalfa than when grown alone, regardless of N application level. When grown alone, alfalfa's N2 fixation was reduced with N fertilization(R2=0.9376, P=0.0057). When grown in a mixture with smooth bromegrass, with 75 kg ha–1 of N fertilizer, the percentage of atmospheric N2 fixation contribution to total N in alfalfa(%Ndfa) had a maximum of 84.07 and 83.05% in the 2nd and 3rd harvests, respectively. Total 3-harvest %Ndfa was higher when alfalfa was grown in a mixture than when grown alone(shoots: |t|=3.39, P=0.0096; root: |t|=3.57, P=0.0073). We believe this was due to smooth bromegrass being better able to absorb available soil N(due to its fibrous root system), resulting inlower soil N availability and allowing alfalfa to develop an effective N2 fixing symbiosis prior to the 1st harvest. Once soil N levels were depleted, alfalfa was able to fix N2, resulting in the majority of its tissue N being derived from biological nitrogen fixation(BNF) in the 2nd and 3rd harvests. When grown in a mixture, with added N, alfalfa established an effective symbiosis earlier than when grown alone; in monoculture BNF did not contribute a significant portion of plant N in the N75 and N150 treatments, whereas in the mixture, BNF contributed 17.90 and 16.28% for these treatments respectively. Alfalfa has a higher BNF efficiency when grown in a mixture, initiating BNF earlier, and having higher N2 fixation due to less inhibition by soil-available N. For the greatest N-use-efficiency and sustainable production, grass-legume mixtures are recommended for improving grasslands, using a moderate amount of N fertilizer(75 kg N ha–1) to provide optimum benefits.     

西北地区制种玉米产量及氮素吸收对供氮水平的响应

【目的】针对我国制种玉米氮素吸收与累积规律不明确的问题,研究制种玉米生物量累积、产量形成和氮素吸收对供氮水平的响应,旨在为制种玉米高产高效绿色生产提供理论依据。【方法】以大面积制种的品种组合为试验材料,于2019—2020年开展田间定位试验。采用完全随机区组设计,共设置4个供氮水平,分别为只施底肥对照(CK)、168 kg N·hm^-2、240 kg N·hm^-2和320 kg N·hm^-2,研究不同供氮水平对制种玉米父母本生物量、杂交种产量和氮素吸收累积的影响。【结果】制种玉米父母本生物量累积随供氮水平的提高而提高,产量随供氮水平的提高先增加后保持稳定,N240处理同时实现了较高的产量、氮肥利用率和籽粒氮浓度,两年结果较为一致。N168处理在试验第2年达到较高产量,但氮浓度低于N240处理。母本秸秆及父本整株氮浓度均为高氮处理高于低氮处理;灌浆期母本实现最大生物量的临界氮浓度为15.08 g·kg^-1,收获期母本生物量与氮浓度呈线性相关。各追施氮肥处理的花后生物量两年间均大于花前,且随供氮水平的提...     

接种菌根真菌和氮添加处理对樟子松苗木根际微生态环境的影响

通过研究接种菌根真菌和N添加处理对植物根际土壤胞外酶及其计量特征,以及微生物养分限制的影响,从而探讨全球N沉降背景下菌根真菌对根际微生态环境的调控机制。以1年生樟子松的盆栽菌根苗（简称+M,混合接菌处理）和非菌根苗（简称-M,未接菌处理）为对象,设置4个N添加处理:不施N（0N,0 kg·hm^-2·a^-1）、低氮（LN,15 kg·hm^-2·a^-1）、中氮（MN,30 kg·hm^-2·a^-1）、高氮（HN,60 kg·hm^-2·a^-1）。对比分析接菌和N添加处理对樟子松苗木根际土壤速效养分和胞外酶活性及其计量特征的影响,探究菌根真菌对微生物养分限制调控规律。结果表明:1）不同接菌处理下,随N添加量增加,土壤中碱解N均呈增加的趋势。HN处理下,菌根苗根际土壤有效磷达到最大。2）LN、MN处理下,菌根苗根际土LAP、NAG和ALP酶活性均显著高于非菌根苗。3）MN、HN处理下,菌根苗根际土壤中微生物碳限制和磷限制较非菌根苗均显著降低。4）通过PLS-PM路径模型分析发现,接菌处理正效应作用于根际土壤微生物养分限制。综上,N添加改变了根际土壤N平衡,菌根真菌有效调控N添加下根际微生态环境的稳定性,降低苗木根际微生物的碳和磷限制。     

优化根际固氮菌种改善水稻生育状况的研究

为了探讨接种不同根际联合固氮菌对水稻生长发育的影响,对水稻接种了不同菌种,试验研究结果表明,接种根际联合固氮菌对水稻分蘖发生发展与成穗有一定的促进作用,对水稻农艺性状有一定的改善作用,稻谷经济产量和生物量比对照增加。稻谷用菌液浸种,水稻发报数比对照略有增加;苗期喷菌株,对水稻株高、发根量以及鲜重、干重有一定的增加作用,秧苗素质有所提高。