四种植物根际促生菌筛选及生物菌肥效果研究

发布时间：2018-05-31 21:58

本文选题：植物根际促生菌(PGPR) + 生物菌肥　；参考：《甘肃农业大学》2017年硕士论文

【摘要】：生物菌肥是一种新型的环境友好型肥料,其内所含的促生菌等有益微生物—植物根际促生菌(Glant Growth Promoting Rhizobacteria,PGPR)不仅能给植物提供营养、降低病虫害发生,对植物起到增产的作用,而且还能改善土壤质量,防止其二次污染。目前,PGPR已成为生物菌肥研究的热点之一。然而,由于植物种类颇多,不同植物中PGPR不同,且其促生效果各异。因此,PGPR菌株资源库还有待进一步完善。为丰富PGPR菌株资源库,了解PGPR菌对不同植物的促生效果,本研究从花椰菜(Brassica oleracea)、油菜(Brassica campestris)、大葱(Allium fistulosum)、辣椒(Capsicum annuum)4种植物根际分离和筛选了PGPR菌,根据菌株形态特征及16S rDNA基因序列分析方法对优良菌株进行鉴定。利用“牛津杯法”测定优良PGPR菌株间的拮抗性,从而获得了最佳的菌株组合。为验证最佳菌株组合的效果,本研究设置不施肥(CK)、前期研制的菌肥(NF1)、新制作的菌肥(NF2)、化肥4个处理进行盆栽试验,测定了小黑麦(Triticosecale)和猫尾草(Phleum pratense)的生长指标。另外,设置A(不施肥,CK)、B(施微生物肥料)、C(70%化肥+微生物肥料)、D(100%化肥)4个处理进行田间试验,研究其对青梗花椰菜(Brassica oleracea)生物指标和土壤微生物学指标的影响。目前取得的结果如下:(1)从4种植物根际中共获得57株PGPR菌株,其中固氮菌24株,溶磷菌33株。(2)PGPR菌株固氮酶活性在0.14~3664.97 nmol(C2H4)/h/mL,NCRS1、PYRS1、NYRS2及PCHP1菌株固氮酶活性大于100 nmol(C2H4)/h/mL,其值依次为3664.97、1557.63、160.03、108.90 nmol(C2H4)/h/mL,其余菌株固氮酶活性均小于100nmol(C2H4)/h/m L。(3)溶解无机磷PGPR菌株的D/d值在1.25~2.73,溶磷量在36.99~362.60μg/mL,各菌株培养液pH在4.57~5.75,其中溶磷量最大是NCRP2菌株;溶解有机磷PGPR菌株的D/d值在1.22~2.81,溶磷量在5.62~55.21μg/mL,各菌株培养液pH在5.89~7.85,其中溶磷量最大是PPRS3菌株。能同时溶解无机磷和有机磷的菌株有NYRS3、NCRS1、NPRS1、MBHP1、MPHP1、MCRP1、MYRS2、MCRP2、MCRS1、MC RS4、PCHP1、PYRS2、PYRS1、PCRP1、PCRS2、PBRP1、PCRS1、PPHP1。(4)PGPR菌株分泌IAA量均小于4μg/mL,NCRP2和NCRP3菌株对小麦长蠕孢(Helminthosporium tritici-vulgaris)、番茄早疫病(Alternariasolani)、黄瓜枯萎病(Fusarium oxysporumn)、马铃薯立枯丝核(Rhizoctonia solani)、油菜菌核(Sclerotiniasclerotiorum)、玉米小斑病(Bipolaria maydis)均有抑制作用,其中NCRP2菌株抑菌率分别是61.23%、53.41%、51.52%、53.16%、77.44%、59.87%,NCRP3菌株抑制率分别是66.44%、52.63%、52.44%、62.27%、67.48%、57.02%。(5)通过对21株优良PGPR进行鉴定,结果发现,PYRS1菌株为Pseudomonas syringae,NCRS1和PBRS2为Pseudomonas migulae;PCRP1菌株为Pseudomonas brassicacearum,MPHP1菌株为Pseudomonas thivervalensis,MCRS1、PCRS2、MC RP1、PYRS2及PPRS3菌株为乙酸钙不动杆菌(Acinetobacter calcoaceticus),MC RS4菌株为Advenella kashmirensis,NCRP1菌株为苦马豆根瘤菌(Rhizobium sphaerophysae),NCRP2、NCRS2菌株为叶杆菌(Phyllobacterium ifriqiyense),NYRS3菌株为Microbacterium binotii;NBRP2菌株为Microbacterium profundi,MBHP1、MYRS2、NYRS2、MCRP2及PCHP1菌株为短小芽胞杆菌(Bacillus pumilus)。(6)盆栽试验结果表明,与NF1相比,NF2处理使小黑麦茎粗和地上生物量鲜重分别增加了6.79%和0.89%,猫尾草茎粗和叶绿素含量分别增加了33.74%和13.72%,不同处理间没有显著性差异(P0.05);NF2处理下小黑麦总根长、根表面积、根体积最大,分别为217.94 cm、11.78 cm2、0.35 cm3,且对0.5~1.0 mm小黑麦根长有显著性影响(P0.05)。不同处理对猫尾草根系形态和不同根直径的总根长没有显著性影响(P0.05),但有一定的促进作用。(7)通过田间试验发现,不同施肥处理下土壤微生物数量、微生物量均随土层的加深而降低,土壤微生物数量为细菌放线菌真菌。其中,处理C效果最佳,与处理D相比,青梗花椰菜株高、茎粗、叶片数、地上鲜重、地上干重、地下鲜重、地下干重、细菌数量、放线菌数量、微生物总数、土壤微生物量碳、土壤微生物量氮和土壤微生物量磷分别增加了6.82%、11.53%、11.76%、9.68%、33.33%、62.5%、33.33%、34.04%~37.61%、8.42%~15.87%、32.53%~33.86%、13.56%~18.6%、3.99%、12.81%~17.99%,真菌数量减少了26.23%~32.89%。不同施肥处理下土壤微生物量与微生物数量存在一定程度的正相关关系。因此,70%化肥+NF2对青梗花椰菜生长具有良好的促进效果。
[Abstract]:Biological microbial fertilizer is a new kind of environment-friendly fertilizer, which contains beneficial microbes, such as Glant Growth Promoting Rhizobacteria, PGPR, not only can provide nutrients to plants, reduce the occurrence of pests and diseases, increase yield to plants, but also improve soil quality and prevent secondary pollution. At present, PGPR has become one of the hot spots in biological bacterial fertilizer research. However, because of the variety of plant species, different plants have different PGPR and different effects on promoting growth. Therefore, the resource library of PGPR strain still needs to be further improved. In order to enrich the PGPR strain resource pool and understand the effect of PGPR bacteria on the growth promoting effect of different plants, this study from the cauliflower (Brassica olerace). A), campestris, Brassica campestris, Allium fistulosum, and pepper (Capsicum annuum) were isolated and screened from the rhizosphere of 4 plants. The fine strains were identified according to the morphological characteristics of the strains and the sequence analysis method of 16S rDNA gene. The best strains were obtained by using the "Oxford Cup" method to determine the antagonism among the excellent PGPR strains. In order to verify the effect of the best strain combination, the study was set up without fertilization (CK), the pre developed bacterial manure (NF1), the newly produced bacterial manure (NF2), 4 treatments of chemical fertilizer, and the growth index of the small rye (Triticosecale) and the Phleum pratense. In addition, the A (no fertilizer, CK), B (microbiological fertilizer), C (70% chemical) were set up. Fertilizer + microbial fertilizer), D (100% chemical fertilizer) 4 treatments were carried out in field experiments to study the effects on biological indexes of broccoli (Brassica oleracea) and soil microbiological indexes. The results were as follows: (1) 57 PGPR strains were obtained from 4 plant rhizosphere, including 24 Azotobacter, 33 phosphate solubilizing bacteria and (2) PGPR strain of nitrogen fixation enzyme. In 0.14~3664.97 nmol (C2H4) /h/mL, NCRS1, PYRS1, NYRS2 and PCHP1, the activity of nitrogenase was greater than 100 nmol (C2H4) /h/mL. The culture solution of each strain was pH in 4.57~5.75, and the maximum amount of phosphorus dissolved was NCRP2; the D/d value of the dissolved organophosphorus PGPR strain was 1.22~2.81, the amount of phosphorus dissolved in 5.62~55.21 um g/mL, the pH in the culture liquid of each strain was 5.89~7.85, and the largest amount of phosphorus dissolved was the PPRS3 strain. MCRP1, MYRS2, MCRP2, MCRS1, MC RS4, PCHP1, PYRS2, PYRS1, PCRP1, PCRS2, PBRP1, (PBRP1) strains are less than 4 micron, tomato early blight, cucumber fusarium wilt disease, potato riser kernel Solani), Brassica sclerotiorum (Sclerotiniasclerotiorum) and corn speckle disease (Bipolaria maydis) had inhibitory effects. The inhibitory rates of NCRP2 strains were 61.23%, 53.41%, 51.52%, 53.16%, 77.44%, 59.87%, and NCRP3, respectively, 66.44%, 52.63%, 52.44%, 62.27%, 67.48%, 57.02%. (5), identified by a good PGPR of 21 strains. The results were found, The strain of PYRS1 is Pseudomonas syringae, NCRS1 and PBRS2 are Pseudomonas migulae, PCRP1 strain is Pseudomonas brassicacearum, MPHP1 strain is Pseudomonas. Rhizobium sphaerophysae, NCRP2, NCRS2 strain were Phyllobacterium ifriqiyense, NYRS3 strain was Microbacterium binotii, and NBRP2 strain was Microbacterium profundi. Treatment made the crude and aboveground biomass fresh weight of small rye increased by 6.79% and 0.89% respectively. The crude and chlorophyll content increased by 33.74% and 13.72%, respectively, and there was no significant difference between different treatments (P0.05). The total root length, the root surface area and the root volume under NF2 treatment were 217.94 cm, 11.78 cm2,0.35 cm3 respectively, and 0.5~1.0 mm respectively. The root length of Triticale had significant effect (P0.05). Different treatments had no significant influence on the root morphology and the total root length of different root diameter (P0.05), but it had a certain promotion effect. (7) through field experiments, it was found that the amount of soil microbes under different fertilization treatments decreased with the depth of soil layer and the amount of soil microorganism. Bacteria actinomycete fungi. Among them, treatment of C is the best, compared with the treatment of D, green stem broccoli plant height, stem diameter, leaf number, aboveground weight, ground dry weight, underground fresh weight, underground dry weight, number of bacteria, actinomycetes, soil microbial biomass carbon, soil microbial biomass nitrogen and soil microbial biomass phosphorus increased by 6.82%, 11.53% respectively. 11.76%, 9.68%, 9.68%, 33.33%, 62.5%, 33.33%, 34.04%~37.61%, 8.42%~15.87%, 32.53%~33.86%, 13.56%~18.6%, 3.99%, 12.81%~17.99%, the number of fungi decreased the positive correlation between soil microbial biomass and microbial biomass under different fertilization treatments. Therefore, 70% fertilizer +NF2 had good growth promotion for the growth of broccoli. Enter the effect.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S144

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