花生根际促生复合菌剂对连作花生生理生化指标和根际细菌群落的影响

【目的】利用多功能根际促生菌剂促进花生生长,缓解连作障碍对花生的生长抑制。【方法】从10年连作花生根际土中筛选根际微生物,测定其促生及拮抗能力,并经16S rRNA基因测序确定菌株分类。选取3株功能互补且相互之间无生长抑制的根际促生菌制备微生物复合菌剂,利用发芽及盆栽试验验证微生物复合菌剂的促生效应。利用高通量测序技术对花生根际土壤细菌的16S rRNA基因的V3-V4区进行测序分析。【结果】从连作花生根际中共筛选获得37株具有促生及抑制植物病原菌生长能力的根际促生菌,选取3株制备微生物复合菌剂。与空白对照相比,复合菌剂显著提升花生发芽率13.22%;与单独使用根际促生菌相比,复合菌剂显著提升花生发芽率分别为6.99%、7.51%、8.87%。施用复合菌剂对花生根系形态、根瘤数量、叶绿素含量、植株光合参数和植株抗氧化酶活均有显著促进作用,花生根系总长、根尖数、主根直径、根系体积和根系活力分别增加了43.50%、49.31%、15.11%、16.92%和112.16%;花生苗期、花针期叶片叶绿素含量和光合作用均得到显著的提高;花生根瘤数量每株增加34个。施用复合菌剂后对花生的根际细菌多样性影响并不显著,在门水平上的优势菌门变形菌门(Proteobacteria)、放线菌门(Actinobacteriota)和拟杆菌门(Bacteroidota)占70%以上。在属水平上新鞘氨醇菌属(Novosphingobium)和鞘氨醇单胞菌属(Sphingomonas)是优势菌属。【结论】本研究研制的花生根际促生复合菌剂可有效促进花生种子发芽、花生根系生长、提高叶片叶绿素含量以及促进植物光合作用,为缓解花生连作障碍及丰产增收提供有效的技术支持。     

植物根际促生菌的筛选及鉴定

【目的】植物根际促生菌(PGPR)和植物的互作关系往往不稳定,PGPR菌群有可能提高菌株对野外环境的适应性。为此,本文根据PGPR促生机制的多样性,从不同植物根际土壤进行了PGPR的筛选及鉴定。【方法】首先,按照固氮、解磷、解钾、拮抗6种常见病原真菌,同时能在植物根际定殖为基本初筛标准,然后在实验室条件下测定初筛菌株的多项促生能力(PGP),最后通过生理生化试验和16SrRNA基因序列分析对所筛菌株进行鉴定。【结果】从江苏扬州、盐城等地土壤样品筛选出14株PGPR,具有体外抑菌、产NH3、产IAA、产HCN、产嗜铁素、解磷、溶钾、固氮以及产抗生素等促生能力。分类鉴定结果显示:7株属于假单胞菌属(Pseudomonas)、3株属于类芽孢杆菌属(Paenibacillus)、2株为芽孢杆菌属(Bacillus)、1株为布克霍尔德氏菌属(Burkholderia)、1株为欧文氏菌属(Erwinia)。【结论】所筛细菌具有多种促生能力,且能在根际定殖,为进一步构建多功能PGPR广适菌群提供菌株资源。     

植物根际促生菌促生特性研究进展

根际微生物组是决定农作物健康状况的关键因素之一,也是调节农作物与生物和非生物环境相互作用的重要因素。植物根际促生菌(plant growth-promoting rhizobacteria, PGPR)为农作物宿主提供了多种有益作用,通过化学交流以复杂的方式与农作物、土壤相互作用,进而促进农作物生长。本文综述了PGPR对农作物的促生机制、PGPR与农作物的互作及其在农业实践中的应用,并展望了PGPR在农业实践中应用的发展趋势,以期为今后PGPR的应用和研究提供新的思路和理论支撑。     

辣椒根际促生菌的分离筛选及抗病促生特性研究

获得辣椒根际促生菌(Plant growth promoting rhizobacteria,PGPR)并探究其抗病促生特性。采用固氮、无机磷和有机磷培养基从江苏省徐州市采集的辣椒根际土壤中分离筛选根际促生菌株(PGPR),通过形态特征及16S rDNA序列分析进行菌株鉴定,对菌株的固氮、解磷、分泌3-吲哚乙酸(IAA)能力及对4种辣椒病害病原菌抗病能力进行探究。得到13株辣椒PGPR菌株,经鉴定分别属于Bacillus、Pseudomonas、Lelliottia、Siccibacter、Achromobacter、Microbacterium和Paenibacillus;13株PGPR菌株均有固氮功能;其中7株可解有机磷,分别属于Lelliottia、Bacillus、Siccibacter、Microbacterium、Paenibacillus;5株可解无机磷,分别属于Lelliottia、Bacillus、Siccibacter、Pseudomonas;3株具有分泌IAA能力,分别属于Lelliottia、Siccibacter、Bacillus;5株具有抗病能力,分别属于B...     

4株茶树根际促生菌菌株的鉴定及促生作用

【背景】根际促生菌可以促进植物生长、提高植物抗性。茶树根际具有特殊的根土微生物生境,可以获得具促生作用的有益微生物。【目的】探究4株茶树根际促生菌菌株的分类地位及促生作用,筛选优良的根际促生菌菌株。【方法】通过形态、生理生化特征、16S rRNA基因序列同源性比对鉴定4株茶树根际促生菌,采用钼锑抗比色法测定溶磷量,通过比色法测定ACC脱氨酶活性、CAS法测定产铁载体能力、Salkowski法测定产IAA (Indoleacetic acid)的能力进行促生作用研究,通过盆栽实验测试白菜、空心菜、苋菜及水稻的株高及鲜重以分析促生效应。【结果】鉴定KKS-6-N1为放射型土壤杆菌(Agrobacteriumradiobacter), KKS-7-N7为铜绿假单胞菌(Pseudomonas aeruginosa),GD3为Pseudomonashunanensis,GD12为弯曲芽孢杆菌(Bacillusflexus)。固氮菌株KKS-6-N1可产铁载体;固氮菌株KKS-7-N7具有解磷及产铁载体能力,分泌的IAA含量高达101.29mg/L;解钾菌株GD3具溶磷能力,分泌的ACC脱氨酶酶活为8.09μmol/(mg·h),相对铁载体含量为0.31;具固氮解钾性能的菌株GD12分泌的ACC脱氨酶活性为14.46μmol/(mg·h)。盆栽试验表明,4个菌株对白菜、空心菜、苋菜的株高和鲜重均有明显促进作用,尤以GD3效果更甚。【结论】茶树根际促生菌菌株Pseudomonas hunanensis GD3促生作用显著,具有开发成微生物菌肥的潜力。     

接种促生菌对花生根际土壤微生物及营养元素的影响

植物根际促生菌是一类可促进植物生长的有益细菌,有效的根际促生菌剂可以减少化肥施用。以束村氏菌属(Tsukamurella sp.)P9、伯克霍尔德氏菌属(Burkholderia sp.)P10、以及P9和P10混合菌液作为接种菌株,研究促生菌对花生生长、植株及土壤营养、根际土壤微生物类群及功能的影响。30 d盆栽实验结果表明,接种组的花生鲜重、株高及根长均显著提高;根际土壤细菌总数、固氮菌和溶磷菌数均明显高于未接种组;氮循环功能菌群数量有不同程度提高,土壤蔗糖酶、脲酶及过氧化氢酶均高于对照;土壤碱解氮及速效钾显著提高,植株营养指标有所提升,尤以P10接种效果更优。本研究初步结论表明2株促生菌通过活化土壤微生物、提高植株的有效营养元素含量,促进了花生的生长。     

玉米根际高效溶磷菌的筛选、鉴定及促生效应研究

为获得玉米根际高效溶磷促生菌(Plant growth-promoting rhizobacteria,PGPR)并明确其促生特性,采用选择培养方法从玉米根际土壤筛选出优良PGPR 菌株,测定其溶磷及分泌吲哚乙酸(IAA)的能力,并对优良菌株进行鉴定;采用盆栽试验研究菌株的促生作用。结果分离到2 株优良PGPR 菌株CH07和FD11,其溶磷量分别为368.5 mg/L和321.5 mg/L,产IAA量分别为30.93 mg/L和15.93 mg/L。形态学特征、生理生化特征和16S rDNA 序列分析结果表明,CH07为芽孢杆菌属(Bacillus aryabhattai),FD11为链霉菌属(Streptomyces maritimus)。最后通过盆栽试验对这2株细菌分离物的促生效果进行比较,结果发现,CH07、FD11,尤其是CH07与FD11的复合物,对苋菜的株高及地上部鲜重有积极作用,可作为研制生物肥料的优良菌株资源。     

一株黄瓜根际促生菌的筛选、鉴定及其发酵特性

Salkowski比色法评价菌株发酵产吲哚乙酸(IAA)的能力;采用平皿、盆栽方法检测菌株的促生能力;对典型菌株生理生化测定及16S rRNA基因序列系统发育分析,初步确定菌株的分类地位;进一步采用正交设计探索不同碳源、氮源对菌株产IAA的影响。结果表明从黄瓜植株根部分离得到菌株18株,其中8株为产IAA菌株。菌株SGM7产IAA能力最强,产量达23.59 mg/L;1%SGM7菌悬液对盆栽黄瓜幼苗有明显促生效果(P0.05);初步鉴定SGM7为短小芽孢杆菌(Bacillus pumilus);其最适产IAA发酵培养基配方为:蛋白胨1%、玉米粉2%、麸皮0.25%、硫酸铵0.05%、硝酸钾0.05%;其发酵液IAA产量高达35.87 mg/L。     

樱桃根际促生细菌的筛选与鉴定

【目的】筛选樱桃[Cerasus pseudocerasus(Lindl.)G.Don]生物肥料中应用的优良菌株资源。【方法】通过保绿法和萝卜子叶增重法,结合定量检测细菌分离物产植物激素的能力,从樱桃根际土壤中筛选具有良好应用潜力的细菌分离物。利用盆栽试验,验证其促生效果。对促生效果较好的细菌分离物,进行形态学观察、生理生化测定、16S rRNA基因序列及系统发育树分析以确定其分类地位。【结果】筛选出4株具有良好应用潜力的细菌分离物,盆栽试验结果表明:AI5、AI21、PII17、PI7,尤其是PII17,显著提高樱桃根系及地上部生物量,对樱桃生长有明显的促进作用。分类鉴定结果显示:AI5、AI21为短小芽孢杆菌(Bacillus pumilus),PII17为假单胞菌属(Pseudomonas sp.),PI7为肠杆菌属(Enterobacter sp.)。【结论】AI5、AI21、PII17、PI7,尤其是PII17,对樱桃生长有明显的促进作用,在生物肥料的生产中具有广阔的应用前景。     

植物根际促生菌的筛选及其对玉米的促生效应

[目的]以不同植物根及根际土壤为研究材料,进行植物根际促生菌(PGPR)的筛选,并探索其植物促生作用机制.[方法]以解磷、固氮、产氨、产IAA和拮抗3种常见病原真菌为筛选标准,测定了初筛菌株的多项促生能力,并通过对这些菌分别单独回接和多菌混接的玉米盆栽试验,测定了其对玉米的促生效应.[结果]从渭南、成阳、安康、商洛和榆林5地分离得到的158株菌中有17株茵具有上述多种植物促生作用的菌株.盆栽试验的测定结果表明:单独接种和多菌混合接种在玉米株高、根长、茎长、茎平均直径和干重方面与对照组相比较都有所增加,尤其是在多个指标上,多菌混合接种所显示出的促生效应均明显优于单菌接种.[结论]所筛选到的具有多种促生能力的菌株,可以为进一步构建植物根际促生菌(PGPR)菌群提供良好的种质资源.     

Identification of rhizobacteria from maize and determination of their plant-growth promoting potential

During the growing season of 1986, the rhizobacteria (including organisms from the ectorhizosphere, the rhizoplane and endorhizosphere) of 20 different maize hybrids sampled from different locations in the Province of Quebec were inventoried by use of seven different selective media. Isolates were characterized by morphological and biochemical tests and identified using the API20E and API20B diagnostic strips.Pseudomonas spp. were the prominent bacteria found in the rhizoplane and in the ectorhizosphere.Bacillus spp. andSerratia spp. were also detected, but in smaller numbers. In the endorhizosphere,Bacillus spp. andPseudomonas spp. were detected in order of importance. Screening for plant growth-promoting rhizobacteria was carried out in three soils with different physical and chemical characteristics. The results depended on the soil used, but two isolates (Serratia liquefaciens andPseudomonas sp.) consistently caused a promotion of plant growth.Contribution no. 350 of the Research Station, Agriculture Canada, Sainte-Foy, Quebec.     

连作花生田根际土壤优势微生物的分离和鉴定

【目的】从不同连作年限的花生田根际土壤中分离优势微生物并进行鉴定,为研究花生连作后优势微生物的变化奠定基础。【方法】采用土壤稀释分离法从不同连作年限花生根际土壤中分离优势细菌、真菌和放线菌,结合菌株形态特征、培养性状、生理生化特征及16S rDNA序列分析对细菌、放线菌进行鉴定,通过形态特征、培养特征和分子鉴定方法对优势真菌进行鉴定。【结果】从连作花生田根际土壤中分离鉴定出7种优势细菌、7种优势真菌和7种优势放线菌。7种优势细菌分别为Leifsonia xyli、氯酚节杆菌(Arthrobacterchlorophenolicus)、黄色微杆菌(Microbacterium flavescens)、鞘氨醇单胞菌属(Sphingomonas sp.)、巴斯德菌属(Pasteurella sp.)、简单芽孢杆菌(Bacillus simplex)和巨大芽孢杆菌(Bacillus megaterium)。7种优势真菌分别为枝状枝孢菌(Cladosporium cladosporioides)、产紫青霉(Penicillium purpurogenum)、哈茨木霉有性型(Hypocrea lixii)、Exophiala pisciphila、微紫青霉(Penicillium janthinellum)、曲霉(Aspergillus sp.)和大丽轮枝菌(Verticillium dahliae)。7种优势放线菌分别为紫红链霉菌(Streptomyces violaceoruber)、华丽黄链霉菌(Streptomyces flaveus)、Streptomyces panaciterrae、不产色链霉菌(Streptomyces achromogenes)、假浅灰链霉菌(Streptomyces pseudogriseolus)、纤维素链霉菌(Streptomyces cellulosae)和金色链霉菌(Streptomyces aureus)。【结论】本研究是第一次系统的从连作花生根际土中分离鉴定优势微生物,种植花生后根际土壤中优势微生物的种类发生了明显变化,但变化没有规律。     

连作花生土壤中酚酸类物质的检测及其对花生的化感作用

研究了南方红壤区不同连作年限花生土壤中酚酸物质的种类、含量,及其对花生生长的影响。结果表明:连作花生土壤中对羟基苯甲酸、香草酸和香豆酸随着连作年限的增加而增加,连作10a后3种酚酸总量达11.09mg·kg-1干土,显著高于连作3a和6a的土壤;而土壤中香豆素和苯甲酸含量比较低,且变化没有规律。所有酚酸处理组对花生幼苗的株高和根长表现出抑制作用,对花生幼苗地下部的干鲜重均表现出"低促高抑"的特点。香草酸和香豆酸处理组对花生幼苗地上部的干鲜重表现出"低促高抑"的特点,其他处理组均表现出抑制作用。花生幼苗根系活力随着酚酸处理浓度的增加而降低,花生幼苗的超氧化物歧化酶活力(SOD)、过氧化物酶活力(POD)、丙二醛含量(MDA)则随着酚酸浓度的增加而增加。与只用茄腐镰刀菌孢子悬液浸泡花生种子的对照相比,加入酚酸后,花生种子的病原菌的感染率随着酚酸浓度的增加而增加,发芽率则随着酚酸浓度的增加而下降。以上结果说明,酚酸物质可以抑制花生幼苗的生长和提高花生的发病率,可能是因为酚酸物质破坏花生细胞膜的完整性而使病原菌入侵,影响花生生长,产生连作障碍。     

Plant-growth promoting effect of newly isolated rhizobacteria varies between two Arabidopsis ecotypes

Various rhizobacteria are known for their beneficial effects on plants, i. e. promotion of growth and induction of systemic resistance against pathogens. These bacteria are categorized as plant growth promoting rhizobacteria (PGPR) and are associated with plant roots. Knowledge of the underlying mechanisms of plant growth promotion in vivo is still very limited, but interference of bacteria with plant hormone metabolism is suggested to play a major role. To obtain new growth promoting bacteria, we started a quest for rhizobacteria that are naturally associated to Arabidopsis thaliana. A suite of native root-associated bacteria were isolated from surface-sterilized roots of the Arabidopsis ecotype Gol-1 derived from a field site near Golm (Berlin area, Germany). We found several Pseudomonas and a Microbacterium species and tested these for growth promotion effects on the Arabidopsis ecotypes Gol-1 and Col-0, and for growth-promotion associated traits, such as auxin production, ACC deaminase activity and phosphate solubilization capacity. We showed that two of the bacteria strains promote plant growth with respect to rosette diameter, stalk length and accelerate development and that the effects were greater when bacteria were applied to Col-0 compared with Gol-1. Furthermore, the capability of promoting growth was not explained by the tested metabolic properties of the bacteria, suggesting that further bacterial traits are required. The natural variation of growth effects, combined with the extensive transgenic approaches available for the model plant Arabidopsis, will build a valuable tool to augment our understanding of the molecular mechanisms involved in the natural Arabidopsis - PGPR association.     

地黄源库关系的变化及其与连作障碍的关系

通过正茬剪叶模拟连作地黄光合源不足条件下,测定分析植株源与库的关系,应用同位素示踪技术分析连作地黄光合产物的分配及内源激素的变化,以探讨地黄的连作障碍机制。结果表明:剪叶处理对植株的影响不能达到连作障碍程度,其库容及其活性大于连作地黄;连作地黄出现光合产物滞留于地上部,库的活性影响了植株光合产物向库的运输,在生长前期植株内源激素发生了变化;因此断定,连作地黄库容及其活性过低是限制地黄块根膨大的重要因素,库通过不同激素对源的反馈调节,抑制了地上源的光合能力及光合产物向库的分配,导致连作障碍。     

滁菊连作土壤中尖孢镰刀菌的分离、鉴定及变化特征

采用传统平板培养法及尖孢镰刀菌专性培养基对滁菊连作土壤的真菌和尖孢镰刀菌进行分析测定,结果表明,在滁菊连作条件下,土壤中真菌和尖孢镰刀菌的数量显著增加,平均分别达到8.71×105 cfu/g干土和3.12×104cfu/g干土,各自是对照土壤的2.24倍和5.57倍。土壤中尖孢镰刀菌数量显著增加可能是导致滁菊连作障碍的重要因素。     

Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat

AIMS: Plant growth promoting rhizobacteria (PGPR) are commonly used as inoculants for improving the growth and yield of agricultural crops, however screening for the selection of effective PGPR strains is very critical. This study focuses on the screening of effective PGPR strains on the basis of their potential for in vitro auxin production and plant growth promoting activity under gnotobiotic conditions. METHODS AND RESULTS: A large number of bacteria were isolated from the rhizosphere soil of wheat plants grown at different sites. Thirty isolates showing prolific growth on agar medium were selected and evaluated for their potential to produce auxins in vitro. Colorimetric analysis showed variable amount of auxins (ranging from 1.1 to 12.1 mg l-1) produced by the rhizobacteria in vitro and amendment of the culture media with l-tryptophan (l-TRP), further stimulated auxin biosynthesis (ranging from 1.8 to 24.8 mg l-1). HPLC analysis confirmed the presence of indole acetic acid (IAA) and indole acetamide (IAM) as the major auxins in the culture filtrates of these rhizobacteria. A series of laboratory experiments conducted on two cv. of wheat under gnotobiotic (axenic) conditions demonstrated increases in root elongation (up to 17.3%), root dry weight (up to 13.5%), shoot elongation (up to 37.7%) and shoot dry weight (up to 36.3%) of inoculated wheat seedlings. Linear positive correlation (r = 0.99) between in vitro auxin production and increase in growth parameters of inoculated seeds was found. Based upon auxin biosynthesis and growth-promoting activity, four isolates were selected and designated as plant growth-promoting rhizobacteria (PGPR). Auxin biosynthesis in sterilized vs nonsterilized soil inoculated with selected PGPR was also monitored that revealed superiority of the selected PGPR over indigenous microflora. Peat-based seed inoculation with selected PGPR isolates exhibited stimulatory effects on grain yields of tested wheat cv. in pot (up to 14.7% increase over control) and field experiments (up to 27.5% increase over control); however, the response varied with cv. and PGPR strains. CONCLUSIONS: It was concluded that the strain, which produced the highest amount of auxins in nonsterilized soil, also caused maximum increase in growth and yield of both the wheat cv. SIGNIFICANCE AND IMPACT OF STUDY: This study suggested that potential for auxin biosynthesis by rhizobacteria could be used as a tool for the screening of effective PGPR strains.     

蚯蚓调控土壤微生态缓解连作障碍的作用机制

连作障碍不仅严重影响作物产量, 而且会导致土壤生物多样性下降、有益微生物减少及病原菌增加等一系列微生态失衡问题。土壤微生态失衡反作用于植物, 导致植物发生更严重的病害、减产等。作为土壤生态系统工程师, 蚯蚓的取食、掘洞和爬行等活动对土壤微生态具有重要的调控作用, 既可以改善土壤环境, 又可以强化土壤生物群落功能, 有望为缓解作物的连作障碍问题提供一条新途径。本文总结了土壤微生态与土壤功能维持及蚯蚓调控土壤生物作用的研究进展, 在此基础上, 结合蚯蚓对化感物质降解作用的研究, 分析了蚯蚓通过调控土壤微生态缓解作物连作障碍的微生物作用机制的三条途径: 直接调控微生物群落、通过改变化感物质组成以及通过调控土壤动物区系调控微生物群落。蚯蚓对微生物群落的调控可改善失衡的土壤根际微生态, 有效缓解作物连作障碍。