基于高通量测序技术的阳光玫瑰不同砧木根际微生物多样性研究  

Analysis of microbial diversity in rhizosphere soil of Shine Muscat grape on different rootstocks using high-throughput sequencing

作　　者：肖慧琳[1] 王建萍[1] 杨业凯 郑秋玲[1] 徐维华[1] 宫磊 宋志忠 唐美玲[1] 刘万好 XIAO Huilin;WANG Jianping;YANG Yekai;ZHENG Qiuling;XU Weihua;GONG Lei;SONG Zhizhong;TANG Meiling;LIUWanhao(Yantai Academy of Agricultural Sciences,Yantai 265500,Shandong,China;College of Horticulture,China Agricultural University,Beijing 100193,China;Shandong Academy of Grape,Jinan 250100,Shandong,China;School of Agriculture,Ludong University,Yan-tai 264025,Shandong,China)

机构地区：[1]山东省烟台市农业科学研究院,山东烟台265500 [2]中国农业大学园艺学院,北京100193 [3]山东省葡萄研究院,济南250100 [4]鲁东大学农学院,山东烟台264025

出　　处：《果树学报》2022年第9期1639-1648,共10页Journal of Fruit Science

基　　金：国家现代农业产业技术体系建设专项资金(CARS-29-17);山东省农业良种工程(2020LZGC008);烟台市科技计划(2020XCZX036);烟台市科技计划(2020XCZX026)。

摘　　要：【目的】探究阳光玫瑰不同砧木和自根苗根际土壤微生物的群落多样性,以期为筛选适宜砧木提供参考。【方法】提取3年生阳光玫瑰嫁接和自根苗木的根际土壤微生物总DNA,用IlluminaMiseq高通量测序技术分析了1103P、5BB、Beta和自根苗土壤根际微生物群落多样性。【结果】1103P显著提高了根际土壤细菌群落的丰度和多样性,Beta显著提高了真菌群落的丰度和多样性,降低了细菌群落的多样性。4个样品细菌OTU归类到44门133纲345目536科1056属,优势细菌门均为变形菌门(Proteobacteria,24.15%~33.57%)、酸杆菌门(Acidobacteriota,14.83%~~22.82%)、放线菌门(Actinobacteriota,7.24%~10.99%),但优势细菌属组成有较大差异,细菌属水平主坐标分析表明,1103P和5BB明显改变了阳光玫瑰根际土壤细菌群落组成,Beta和自根苗细菌群落组成相似度高。4个样品真菌OTU归类到8门33纲82目188科412属,优势真菌门均为子囊菌门(Ascomycota,55.57%~64.86%)、毛霉菌门(Mucoromycota,5.84%~22.24%)和担子菌门(Basidiomycota,12.72%~19.22%),但优势真菌属组成有较大差异,真菌属水平主坐标分析表明,4个样品真菌群落组成差异较大。对细菌群落(门水平)和土壤理化性质的冗余分析结果表明,有机质含量(OM)、交换性镁(Mg^(2+))含量和pH值对根际土壤优势细菌门组成影响较大。【结论】砧木能够改变阳光玫瑰根际土壤微生物群落组成和数量,为筛选适宜砧木和改善根际微生物群落组成提供理论依据。【Objective】In order to provide reference for rootstock screening and cultivating, the community diversity of rhizosphere soil microorganisms of different rootstocks and self-rooted seedlings of Shine Muscat grape was explored. Rhizospheric microorganisms live at the interface between plant roots and soil and interact with plants. Beneficial rhizosphere microorganisms help plants to obtain nutrients and improve plant resistance to abiotic stresses. Harmful rhizosphere microorganisms compete with plants for nutrients in the soil or infect plants through roots, which further inhibits the healthy growth of plants. Plant roots and soil are two main factors affecting microbial community.【Methods】Three-year-old Shine Muscat grape was used as the test material. There were three kinds of rootstock,including 1103P, 5BB and Beta, and self-rooted seedlings served as the control. Soil organic matter was determined by the potassium dichromate volumetric-dilution heat method. pH value was measured by 1∶1water soil ratio and a pH meter. Alkaline hydrolysis of nitrogen was analyzed using the alkaline hydrolysis diffusion method. The available phosphorus was extracted by NaHCO3and further determined by the molybdenum antimony sulfate anti colorimetry. Soil available potassium was extracted by ammonium acetate and then determined by the flame spectrophotometry. Exchangeable calcium and magnesium were extracted with 1 mol · L-1NH4OAc solution, and the content of soluble calcium and magnesium was determined by the atomic absorption spectrophotometer. The total DNA of rhizosphere soil microorganisms was extracted using the E.Z.N.A.? Soil DNA Kit(Omega Bio-tek, Norcross, GA, U.S.A.). The amplification primers of soil bacteria were 341F(5’-CCTAYGGGRBGCASCAG-3’) and 806R(5’-GGACTACNNGGGTATCTAAT-3’), and the amplification primers of soil fungi were ITS1F(5’-CTTGGTCATTTAGAGGAAGTAA-3’) and ITS2R(5’-GCTGCGTTCTTCATCGATGC-3’). PCR reaction conditions were followed below: pre-denaturation at 95 ℃ for 5 min;denaturat

关 键 词：葡萄砧木 根际微生物 高通量测序 

分 类 号：S663.1[农业科学—果树学]