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基于根际营养利用能力的植物根际促生菌的筛选及促生效应研究

发布时间:2018-08-09 18:01
【摘要】:优良菌种是微生物肥料的基础,根际竞争能力是微生物肥料菌株发挥作用的前提,筛选的植物根际促生菌必须要能较好地适应于植物的根际环境。本研究利用玉米、黄瓜、番茄三种作物的根系分泌物为初筛培养基的营养源,从相对应作物的根际土壤样品中分离筛选能利用根系分泌物快速生长的菌株,通过测定所筛菌株的促生特性,进一步结合平皿幼苗促生试验复筛优良菌株,并通过盆栽试验评价促生效果及对所筛菌株进行系统鉴定。最后选择JScG、SCmB、XJmG、SDtG四株芽孢杆菌进行针对不同作物的根际定殖能力研究、液体发酵培养基组分优化、根际促生微生物肥料研制并研究其盆栽促生效果。所获主要结果如下:1.PGPR菌株初筛共获得24株菌株,它们均可在以植物根系分泌物为唯一营养来源的培养基上迅速生长,且具有溶磷、产NH3、产IAA及产嗜铁素等一种或多种促生性能。其中具有产氨能力的14株,占所筛菌株的58.3%;可产IAA的20株,占所筛菌株的83.3%,菌株JScB的IAA产量最大,达到了 73.28 mg L-1;可产铁载体的12株,占所筛菌株的50%;具有溶解有机磷能力的15株,占所筛菌株的62.5%;所筛各菌株均有一定的溶解无机磷的能力,但差异很大,溶磷量范围在19.14-200.05 mg L-1。根据平皿幼苗促生试验,最终复筛出12株PGPR菌株。其中对玉米有促生作用的根际细菌为SCmB、SCmD、NMGmA、XJmG;对番茄有促生作用的根际细菌为SDtC、SDtG、GZtB、GZtE;对黄瓜有促生作用的根际细菌为HNcB、HNcH、JScB、JScG。2.将复筛得到的12株PGPR菌株用于盆栽试验,分别回接至宿主植物玉米、黄瓜、番茄根际,生长40d后其株高、生物量、根系形态(总根长、根表面积等)和叶绿素含量等显著高于不接菌对照,经16SrDNA分子鉴定,5株为芽孢杆菌属(Bacillus spp.);3株为剑菌属(Ensifer spp.);2株为根瘤菌属(Rhizobium spp.);1株为苍白杆菌属(Ochrobactrum sp.);1株为微杆菌属(Microbacterium sp.)。其中,在玉米盆栽试验中,菌株SCmB促生效果最好,其次为XJmG;在番茄盆栽试验中,促生效果最好的菌株为GZtB,其次为SDtG;在黄瓜盆栽试验中,促生效果最好的菌株为HNcB,其次为JScB和JScG。上述促生效果好的7株菌中,有4株是芽孢杆菌,能产生耐热抗逆的芽孢,是根际促生微生物肥料研发的理想菌种,故选择SCmB、XJmG、SDtG、JScG四株芽孢杆菌进行深入研究。3.测定了 JScG、SCmB、XJmG、SDtG四株芽孢杆菌的生物膜形成及对玉米、黄瓜、番茄三种作物的根际定殖能力。其中菌株SDtG和JScG的生物膜形成能力最强,SCmB次之,XJmG最弱。在限菌砂培定殖试验中,SCmB菌株在玉米根际的定殖能力最强,达到了 9.29×106CFUg-1根;SDtG和JScG在黄瓜根际的定殖能力最强,分别达到了 4.69×106CFU g-1根和4.12×106CFU-1根;XJmG在番茄根际的定殖能力最强,达到了 8.07× 1 06 CFU g-1 根。4.利用单因素试验和正交试验对四株芽孢杆菌的液体发酵培养基组分进行了优化,菌株JScG和SCmB液体发酵培养基的最优配比为糖蜜3%,复合氮源0.6%,氯化钠0.5%,氯化钙0.1%,硫酸锰0.05%;XJmG最优配比为糖蜜2.5%,复合氮源0.6%,氯化钠0.5%,磷酸二氢钾0.1%,氯化钙0.1%,硫酸镁0.1%,硫酸锰0.05%;SDtG最优配比为糖蜜3%,复合氮源0.6%,氯化钠0.25%。5.根据四株芽孢杆菌在液体发酵培养基中的生长曲线,选择了 72 h的液体发酵液进行了根际促生微生物肥的初步制作,即用腐熟堆肥吸附10%(v/w)液体发酵液制作四种微生物肥料,其芽孢数在5.0×108 CFU g-1干重,施用0.5%的微生物肥料于玉米和番茄根际,生长35 d后其株高、生物量、根系形态(总根长、根表面积等)等显著高于对照,其中菌株SCmB对玉米的促生效果最好,菌株XJmG对番茄的促生效果最好。
[Abstract]:The excellent strain is the basis of microbial fertilizer, and the competitive ability of the rhizosphere is the prerequisite for the microbial fertilizer strain to play its role. The selected plant rhizosphere bacteria must be well adapted to the rhizosphere environment of the plant. This study uses the root secretions of three crops of maize, cucumber and tomato as the source of the primary culture medium. The strain of rapid growth of root exudates was isolated and screened in the rhizosphere soil samples. By measuring the growth promoting characteristics of the screened strains, the fine strains were screened by the seedling growth promoting test of Petri dish seedlings, and the effect of growth promotion and the system identification of the screened strains were evaluated by pot experiment. Finally, the four buds of JScG, SCmB, XJmG and SDtG were selected. The bacilli carried out the study on the rhizosphere colonization ability of different crops, the optimization of the medium composition of liquid fermentation medium, the development of rhizosphere microbial fertilizer and the effect of its pot cultivation. The main results were as follows: 24 strains of 1.PGPR strains were obtained at the beginning of screening, and they could be used in the culture medium with the sole nutrient source of plant root exudates. It is fast growing and has one or more growth promoting properties, such as phosphorus dissolving, producing NH3, producing IAA and ferric iron, which have 14 strains of ammonia production, accounting for 58.3% of the sieved strain, 20 of IAA and 83.3% of the screened strain, and the IAA yield of the strain JScB is 73.28 mg L-1, and 12 of the ferric carrier, 50% of the screened strain. 15 strains of organophosphorus, which accounted for 62.5% of the screened strains, all had a certain ability to dissolve inorganic phosphorus, but the difference was very large. The amount of phosphorus dissolved in 19.14-200.05 mg L-1. was finally screened out of 12 PGPR strains according to the growth promoting test of Petri dish seedlings. Among them, the Rhizosphere bacteria that have the promoting effect on maize were SCmB, SCmD, NMGmA, XJmG; and tomato. SDtC, SDtG, GZtB, GZtE, the rhizosphere bacteria that have the effect of promoting growth, and the rhizosphere bacteria that have a stimulating effect on cucumber are HNcB, HNcH, JScB, and JScG.2. will be used to resieve 12 strains of PGPR strains used for pot experiment, and the plant height, biomass, root morphology (total root length, root surface area, etc.) of the host plant maize, cucumber, tomato rhizosphere and growth 40d respectively. Chlorophyll content was significantly higher than that of non bacteria control. 5 strains were identified by 16SrDNA molecule (Bacillus spp.), 3 were rapiti (Ensifer spp.), 2 strains were rhizobia (Rhizobium spp.), 1 were pallid (Ochrobactrum sp.), and 1 strains were micro bacilli (Microbacterium sp.). In the pot experiment of maize, strain SCmB In the pot experiment of tomato, the best growth promoting effect was GZtB, followed by SDtG. In the pot experiment of cucumber, the best producing strain was HNcB, followed by 7 strains of JScB and JScG., the 7 strains were bacillus, which could produce the spores with heat resistance and resistance. Four strains of Bacillus, SCmB, XJmG, SDtG, and JScG were selected for further study on the ideal strain of fertilizer research and development. The biofilm formation of JScG, SCmB, XJmG, SDtG four strains of Bacillus and the rhizosphere colonization of three crops of maize, cucumber and tomato were determined by.3.. Among them, the biofilm of SDtG and JScG was the strongest, and the SCmB was the weakest. The colonization ability of SCmB strain in maize rhizosphere was the strongest, reaching 9.29 x 106CFUg-1 roots in maize rhizosphere; SDtG and JScG had the strongest colonization ability in Cucumber Rhizosphere, which reached 4.69 x 106CFU g-1 root and 4.12 x 106CFU-1 root respectively. XJmG had the strongest colonization ability in tomato rhizosphere, reaching 8.07 x 106 CFU g-1 root.4. utilization single. Factors test and orthogonal test were used to optimize the liquid fermentation medium of four Bacillus strains. The optimal ratio of strain JScG and SCmB liquid fermentation medium was molasses 3%, compound nitrogen source 0.6%, sodium chloride 0.5%, calcium chloride 0.1%, manganese sulphate 0.05%, XJmG optimum ratio was sugar honey 2.5%, compound nitrogen source 0.6%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.1% Calcium chloride 0.1%, Magnesium Sulfate 0.1%, manganese sulphate 0.05%, SDtG optimum ratio of molasses 3%, compound nitrogen source 0.6%, sodium chloride 0.25%.5. based on the growth curve of four bacillus in liquid fermentation medium, selected 72 h liquid fermentation liquid to produce rhizosphere microbial fertilizer, that is to use decomposed composting for 10% (v/w) liquid fermentation. Four kinds of microbial fertilizers were made in the liquid, the number of spore was 5 x 108 CFU g-1 dry weight, and 0.5% microbial fertilizer was used in maize and tomato rhizosphere. After the growth of 35 d, the plant height, biomass, root morphology (total root length, root surface area and so on) were significantly higher than those of the control, among which strain SCmB had the best effect on maize growth, and the growth promoting effect of strain XJmG on Tomato Best.
【学位授予单位】:南京农业大学
【学位级别】:硕士
【学位授予年份】:2015
【分类号】:S144;S182

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