生防菌AS_(818)抗药性标记株在大豆根际定殖

通过60 Co 射线 (1 .0× 1 0 4 rad)对生防菌株链霉菌AS81 8孢子悬液 (1 0 6/mL)进行诱变 ,得到抗链霉素 (>5 0μg/mL)突变株 5株。经诱导 ,RL 4抗链霉素水平达到 1 0 0μg/mL。标记株RL 4在 1 %水琼脂 (WA)和无菌土中培养的大豆根际定殖趋势有所不同。RL 4在1 %WA培养的大豆根表的定殖量呈逐步上升趋势。无菌土中种植的大豆根际和根表的检测表明 :RL 4可在无菌土中大豆根际短期定殖 ,在根际第 1周数量降低了 1 0 0倍 ,以后数量开始逐渐上升 ,第 3周达到高峰 ,数量比第 1周增加了 3个数量级 ,但在第 4周其数量又开始下降 ;在根表RL 4数量逐步下降 ,到第 4周已检测不到RL 4的存在。组织印记法检测发现标记株RL 4可以在无菌培养的大豆根表定殖并沿根分布 ,但却不能定殖于根内。

大豆根际促生菌的分离筛选及其对大豆和百脉根生长与品质的影响

从贵州毕节地区大豆根际土壤中分离溶磷菌株,从大豆根瘤中分离根瘤菌株。对分离出的溶磷圈直径与菌落直径的比值(D/d值)在2.20以上的溶磷菌株分别进行溶磷能力、生长素(IAA)及有机酸分泌能力、产酸产碱性能测定,筛选出优良溶磷菌4株;对分离出的根瘤菌通过大豆试管苗回接及促生效应试验,筛选出高效根瘤菌2株。将筛选出的6株菌经拮抗反应试验后分别按单一溶磷菌接种剂、单一根瘤菌接种剂、溶磷菌+根瘤菌复合接种剂3种处理制备菌悬液,采用盆栽方法分别对大豆和百脉根进行促生效应试验。结果表明,与对照相比,除单一根瘤接种剂对增加大豆株高无效果外,另外2个处理对大豆株高有提升效果,所有处理对大豆茎粗、生物量、结荚数、荚重、单粒数及单粒重都有明显的促进作用。其中溶磷菌+根瘤菌复合菌液对大豆株高、茎粗、幼苗地上生物量和地下生物量的促进效果最好,分别比对照高出21.26%,40.79%,15.88%和42.19%。3个处理对百脉根的第一、二茬株高及地上生物量、全氮、全磷和粗蛋白含量均有明显的提升效应,其中依然是溶磷菌+根瘤菌复合菌液的处理效果最好,分别比对照高出20.82%,54.88%,106.14%,148.78%,19.34%,61.88%和19.34%,与对照差异均达极显著水平(P<0.01)。说明所筛选的菌株组合能产生良好的促生互作效应。

四川地区结瘤大豆根际土壤中紫云英、苜蓿和三叶草根瘤菌的多样性分析

为了解四川地区结瘤大豆根际土壤是否存在与紫云英、苜蓿和三叶草共生结瘤的根瘤菌及其多样性,从四川盆地采集不同种植模式下结瘤大豆根际土壤样品31份,利用紫云英、苜蓿和三叶草进行盆栽捕获试验以获得共生根瘤,从根瘤中分离纯化出根瘤菌,对其进行表型和分子鉴定;并将根瘤菌回接到紫云英、苜蓿、三叶草和大豆根际以检测根瘤菌的寄主范围.在捕获实验中,苜蓿和三叶草分别在6个土壤样品中共生结瘤,而紫云英只在2个土样中共生结瘤,并从共生根瘤中分离出14株根瘤菌;16S r DNA基因序列的相似性分析表明这14株根瘤菌均属于根瘤菌属(Rhizobium),且16S r DNA基因的系统发育树揭示它们分属于根瘤菌属的不同种;这14株根瘤菌在回接实验中都只能让捕获豆科植物结瘤,而不能让其他3种豆科植物结瘤.本研究结果表明,四川地区结瘤大豆根际土壤中的紫云英根瘤菌(Rhizobium astragali)、苜蓿根瘤菌(R.meliloti)和三叶草根瘤菌(R.leguminosarum var.trifolii)资源较少,其与大豆根瘤菌(Bradyrhizobium japonicum)分属于不同的属,且根瘤菌的回接实验进一步证明了根瘤菌的寄主专一性.

黑土根际土壤有机碳及结构对长期施肥的响应

由于人类农业措施的干扰,氮肥和畜禽粪污大量输入到黑土中,对土壤有机碳库产生了较大负面影响。根际有机碳在调控土壤碳循环和养分转化中发挥着重要的作用。探明根际有机碳对不同养分的生态响应,可为不同施肥处理下黑土农田生态系统碳固持和农田可持续利用提供理论依据。该研究以黑土长期定位试验为基础,采集长期不施肥(CK)、常量氮(N)、二倍量氮(N2)、常量有机肥(M)、二倍量有机肥(M2)、常量有机肥+常量氮(MN)、二倍量有机肥+二倍量氮(M2N2)7个处理下大豆根际土壤,分析了根际有机碳和活性有机碳特征,同时利用固态核磁共振技术分析其光谱特征。结果表明,N、M2、MN和M2N2处理的根际土壤有机碳含量显著高于非根际水平,且以MN和M2N2处理的根际效应最显著,分别比非根际增加了18.3%和26.7%。分析核磁共振光谱显示,与非根际土壤相比,根际土壤具有较高的烷氧基碳比例和较低芳香碳比例,表明根际效应能够改变土壤有机碳结构比例。与不施肥处理相比,大部分施肥处理提升了黑土根际有机碳含量,其中以氮肥马粪配施和二倍量马粪(M2N2)处理提升幅度最高。由核磁共振图谱可知,M2和M2N2处理均增加根际土壤难降解成分烷基碳比例、芳香基碳比例、烷基碳与烷氧基碳比值、芳香碳与总碳比值,而MN处理仅增加烷基碳比例、烷氧基碳比例以及烷基碳与烷氧基碳比值。二倍量氮肥(N2)处理降低烷基碳比例、芳香碳比例和烷氧基碳比例,根际土壤难降解成分降低,不利于土壤固碳,同时证明固态^(13)C-核磁共振技术结合半定量分析能够准确地分析不同有机碳结构组分变化,深刻认识根际土壤有机碳的稳定机制。

长期不同施肥处理对黑土根际土壤有机碳结构组分的影响

土壤有机碳是农业生态系统的关键驱动和调节者,特别是根际微域有机碳动态对土壤碳素循环和矿质营养元素释放起着重要作用。研究长期不同化肥和有机肥施用下大豆根际土壤有机碳、活性有机碳以及有机碳结构的变化规律,深入了解根际有机碳固持和稳定机制,为完善农田生态系统碳固持和农田可持续发展提供科学依据和理论支撑。该研究依托黑土长期定位试验,采用化学分析、固态^(13)C-核磁共振(^(13)C-NMR)等方法研究大豆根际土壤有机碳含量、活性有机碳含量和有机碳结构组分变化规律。结果表明,与非根际土壤相比,大豆根际土壤有机碳含量显著增加,长期施肥处理能够显著增加根际土壤有机碳和低活性有机碳含量,以常量有机肥加氮磷钾(MNPK)处理提升效果最好。核磁共振实验结果表明,与不施肥处理相比,MNPK处理明显增加根际土壤烷基碳、烷氧基碳比例以及烷基碳/烷氧基碳比值,降低芳香基碳和芳香碳/总碳比值,在非根际土壤中尤其显著;常量氮磷钾(NPK)处理增加芳香基碳比例和芳香碳/总碳比值,在根际土壤中烷基碳比例和烷基碳/烷氧基碳比值增加,烷氧基碳比例降低,非根际土壤测试结果相反。综上所述,MNPK处理能够显著提升根际有机碳含量,增加有机碳中烷基碳、烷氧基碳比例以及烷基碳/烷氧基碳比值,促进团聚体形成和增加土粒结构稳定性,而NPK处理增加芳香基碳比例和芳香碳/总碳比值,降低根际烷氧基碳比例,团聚体稳定性降低,同时证明固态^(13)C-核磁共振技术结合半定量分析能够准确地分析不同有机碳结构组分变化,深刻认识根际土壤有机碳的稳定机制。

浓度梯度凝胶配制的简易装置设计及应用效果

PCR-DGGE是分子生物学工作者分析微生物区系的一个有效的方法,但是较贵的浓度梯度配胶设备限制了该技术的应用,一个简易的装置能够有效地普及这项技术。本文报道了这一简易装置的设计和利用该装置对大豆根化感物质降解细菌的分析效果。

Temporal and Spatial Dynamics of Bacterial Community in the Rhizosphere of Soybean Genotypes Grown in a Black Soil

The dynamics of rhizosphere microbial communities is important for plant health and productivity, and can be influenced by soil type, plant species or genotype, and plant growth stage. A pot experiment was carried out to examine the dynamics of microbial communities in the rhizosphere of two soybean genotypes grown in a black soil in Northeast China with a long history of soybean cultivation. The two soybean genotypes, Beifeng 11 and Hai 9731, differing in productivity were grown in a mixture of black soil and siliceous sand. The bacterial communities were compared at three zone locations including rhizoplane, rhizosphere, and bulk soil at the third node （V3）, early flowering （R1）, and early pod （R3） stages using polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DCGE） of 16S rDNA. The results of principal component analyses （PCA） showed that the bacterial community structure changed with growth stage. Spatially, the bacterial communities in the rhizoplane and rhizosphere were significantly different from those in the bulk soil. Nevertheless, the bacterial communities in the rhizoplane were distinct from those in the rhizosphere at the V3 stage, while no obvious differences were found at the R1 and R3 stages. For the two genotypes, the bacterial community structure was similar at the V3 stage, but differed at the R1 and R3 stages. In other words, some bacterial populations became dominant and some others recessive at the two later stages, which contributed to the variation of the bacterial community between the two genotypes. These results suggest that soybean plants can modify the rhizosphere bacterial communities in the black soil, and there existed genotype-specific bacterial populations in the rhizospbere, which may be related to soybean productivity.

Soil Respiration During a Soy bean-Growing Season

Soil respiration induced by soybean cultivation over its entire growing season and the factors influencing soil respiration were investigated to examine the seasonal pattern of soil respiration induced by soybean cultivation, explore soybean growth and photosynthesis on soil respiration, and determine the temperature dependence on soil respiration. Soil respiration in a pot experiment with and without soybean plants was sampled using the static chamber method and measured using gas chromatograph. Air temperature was a dominant factor controlling soil respiration rate in unplanted soil. Additionally, rhizosphere respiration comprised 62% to 98% of the soil respiration rate in the soybean-planted soil varying with the soybean growth stages. Harvesting aerial parts of soybean plant caused an immediate drop in the soil respiration rate at that stage. After harvesting the aerial parts of the soybean plant, a highly significant correlation between soil respiration rate and air temperature was found at the flowering stage （P 〈 0.01）, the pod stage （P 〈 0.01）, and the seed-filling stage （P 〈 0.05）. Thus, rhizosphere respiration during the soybean-growing period not only made a great contribution to soil respiration, but also determined the seasonal variation pattern of the soll respiration rate.

Dynamics of Microbial Activity Related to N Cycling in Cd-Contaminated Soil During Growth of Soybean

The potential influences of cadmium (Cd) on the biochemical processes of the soil nitrogen (N) cycle, along with the dynamics of ammonification, nitrification, and denitrification processes in the rhizosphere and non-rhizosphere (bulk soil), respectively, were investigated in a Cd-stressed system during an entire soybean growing season. In terms of Cd pollution at the seedling stage, the ammonifying bacteria proved to be the most sensitive microorganisms, whereas the effects of Cd on denitrification were not obvious. Following the growth of soybeans, the influences of Cd on ammonification in the bulk soil were: toxic impacts at the seedling stage, stimulatory effects during the early flowering stage, and adaptation to the pollutant during the podding and ripening stages. Although nitrification and denitrification in the bulk soil decreased throughout the entire growth cycle, positive adaptation to Cd stress was observed during the ripening stage. Moreover, during the ripening stage, denitrification in the bulk soil under high Cd treatment (20 mg kg-1) was even higher than that in the control, indicating a probable change in the ecology of the denitrifying microbes in the Cd-stressed system. Changes in the activity of microbes in the rhizosphere following plant growth were similar to those in the non-rhizosphere in Cd treatments; however, the tendency of change in the rhizosphere seemed to be more moderate. This suggested that there was some mitigation of Cd stress in the rhizosphere.