洛阳市烟田土壤微生物检测与拮抗真菌的筛选

发布时间：2018-12-16 00:03

【摘要】：为了明确洛阳地区烟田土壤微生物的年动态变化以及拮抗真菌的种类,试验以洛阳市嵩县、洛宁与汝阳烟草不同生育期的烟田土壤为样本,用平板稀释法进行土壤微生物的分离,用对峙培养法进行土壤真菌对病原真菌拮抗作用的筛选,依据形态学特征结合ITS序列分析对拮抗菌株进行鉴定,获得了以下试验结果。(1)烟田土壤微生物的数量为细菌真菌放线菌,最大值为1.31×109、3.4×105、2.7×105CFU.g-1;在烤烟不同生育期土壤细菌、真菌数量表现出先增加后减少的趋势,在旺长期达到峰值,以植烟3-6年的土壤细菌年变化最稳定。土壤放线菌数量在不同样本地、不同生育期、不同植烟年限表现各不同。(2)分离保存真菌178株,其中136株属于15个不同属,青霉属、镰孢菌属、曲霉属与木霉属合计分离频率达63.83%,为土壤中的优势种群。(3)筛选出具有拮抗作用的真菌菌株17株,其中木霉菌株5株,青霉菌株5株,曲霉菌株2株,待鉴定菌株5株。(4)具有拮抗作用的木霉菌为哈茨木霉(Trichoderma harzianum Rifai)、绒毛木霉(Trichoderma tomentosum Bissett,KubicekSzakaes)、棘孢木霉(Trichoderma asperellum Samuels)、深绿木霉(Trichoderma atroviride Karsten)和毛簇木霉(Trichoderma velutinum Bissett,KubicekSzakaes)。青霉菌为为产黄青霉(Penicillium chrysogenum)、纯绿青霉(Penicillium polonicum)和青霉菌(Penicillium sp.);曲霉菌为土曲霉(Aspergillus terreus Thom)。(5)哈茨木霉和棘孢木霉对尖孢镰刀菌(Fusarium oxysporum Schlecht)、烟草疫霉(Phytophthora parasitica var.nicotianae(Breda de Hean)Tuker)和瓜果腐霉(Pythium aphanidermatum(Edson)Fitzpatrick)均有较好的拮抗效果,棘孢木霉对以上三种病原真菌均有抑制作用尚属首次报道。毛簇木霉对烟草疫霉和尖孢镰刀菌有较好的拮抗效果。绒毛木霉对烟草疫霉和瓜果腐霉有较好的拮抗效果,在国内系首次报道。(6)青霉菌(Penicillium sp.)R3-3对瓜果腐霉和尖孢镰刀菌的抑菌效果最好;土曲霉(Aspergillus terreus)对烟草疫霉和瓜果腐霉均有较好的拮抗效果。本研究通过对土壤微生物的分离、筛选和鉴定,明确了洛阳地区土壤微生物的年动态变化及拮抗真菌的种类,为生产中了解烟田土壤微生物的构成与变化,开展烟草土传病害生物防治提供了一定的理论与应用价值。
[Abstract]:In order to determine the annual dynamic changes of soil microbes and the species of antagonistic fungi in tobacco fields in Luoyang area, the soil samples from Songxian County, Luoyang City, Luoning and Ruyang tobacco fields at different growth stages were used as samples. Soil microbes were isolated by plate dilution method, antagonistic effects of soil fungi against pathogenic fungi were screened by confrontation culture method, and antagonistic strains were identified according to morphological characteristics and ITS sequence analysis. The following results were obtained: (1) the number of soil microorganism in tobacco field was bacterial fungi actinomycetes, the maximum value was 1.31 脳 10 9 (3.4 脳 10 5N) 2.7 脳 10 5 CFU 路g -1; At different growth stages of flue-cured tobacco, the number of fungi increased first and then decreased, and reached the peak value in the peak period. The annual variation of soil bacteria was the most stable in tobacco growing period from 3 to 6 years. The number of soil actinomycetes in different samples, different growth stages, different years of tobacco planting were different. (2) 178 strains of fungi were isolated and preserved, of which 136 belonged to 15 different genera, Penicillium, Fusarium. The total isolation frequency of Aspergillus and Trichoderma was 63.83, which was the dominant population in soil. (3) 17 strains of fungi with antagonistic effect were screened, including 5 strains of Trichoderma, 5 strains of Penicillium, 2 strains of Aspergillus. Five strains were identified. (4) the antagonistic Trichoderma was Trichoderma (Trichoderma harzianum Rifai), (Trichoderma tomentosum Bissett,KubicekSzakaes (Trichoderma tomentosum Bissett,KubicekSzakaes), Trichoderma sp. (Trichoderma asperellum Samuels), (Trichoderma atroviride Karsten) and Trichoderma tuber (Trichoderma velutinum Bissett,KubicekSzakaes). Penicillium spp. (Penicillium chrysogenum),: pure penicillium (Penicillium polonicum) and penicillium (Penicillium sp.); Aspergillus sp. (Aspergillus terreus Thom). (5) Trichoderma harzii and Trichoderma spinoides had better antagonistic effects against (Fusarium oxysporum Schlecht), (Fusarium oxysporum Schlecht), Phytophthora tabaci, (Phytophthora parasitica var.nicotianae (Breda de Hean) Tuker) and (Pythium aphanidermatum (Edson) Fitzpatrick). The inhibitory effects of Trichoderma spinoides on the above three pathogenic fungi were reported for the first time. Trichoderma tubuloides had a good antagonistic effect against Phytophthora tabacum and Fusarium oxysporum. Trichoderma tomentosa had a better antagonistic effect against Phytophthora tabacum and Pythium melons. (6) Penicillium (Penicillium sp.) R3-3 had the best antimicrobial effect against Pythium fusarium and Fusarium oxysporum; Aspergillus earth (Aspergillus terreus) had a better antagonistic effect against Phytophthora tabacum and Pythium melons. Through the isolation, screening and identification of soil microbes, the annual dynamic changes of soil microbes and the species of antagonistic fungi in Luoyang area were determined in order to understand the composition and changes of soil microbes in tobacco fields. Biological control of tobacco soil-borne diseases provides certain theoretical and practical value.
【学位授予单位】：河南科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.3;S572

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