| 3种竹子内生固氮菌特征及多样性 |
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| 引用本文： | 冯歌林,高竞,严淑娴,王晶,梁辰飞,秦华,陈俊辉,徐秋芳.3种竹子内生固氮菌特征及多样性[J].浙江农林大学学报,2021,38(6):1203-1212. |
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| 作者姓名： | 冯歌林 高竞 严淑娴 王晶 梁辰飞 秦华 陈俊辉 徐秋芳 |
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| 作者单位： | 1.浙江农林大学 环境与资源学院，浙江 杭州 3113002.浙江农林大学 省部共建亚热带森林培育国家重点实验室，浙江 杭州 311300 |
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| 基金项目： | 浙江省自然科学基金重点资助项目(LZ16C160002)；国家自然科学基金资助项目(31570602) |
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| 摘 要： | 目的 了解不同竹子内生固氮菌及其定殖土壤根区固氮菌的特征和生物多样性。 方法 以毛竹Phylloslachys edulis、狭叶青苦竹Pleioblastus chino、孝顺竹Bambusa multiplex叶部、根部及定植土壤为对象，设计nifH固氮基因引物，采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)和随机克隆技术，通过聚类分析、Shannon多样性指数分析构建系统发育树。 结果 不同竹子根区土壤之间、同一竹种根部与叶部之间、不同竹种同一器官之间、根区土壤与植物之间的固氮菌群落结构差异明显，同一竹种叶部或根部的重复样品相似度均不高；根部内生固氮菌种类总体多于叶部，土壤固氮菌与根部内生固氮菌相似度高于叶部。不同竹种不同器官间固氮菌Shannon多样性指数差异显著，孝顺竹叶部(2.73)与土壤(2.70)大于根部(2.53)(P＜0.05)，狭叶青苦竹根部和毛竹根部大于叶部和土壤(P＜0.05)。植物样品随机克隆测序对比得到83株固氮菌，仅12株为可培养固氮菌株，分别属于慢生根瘤菌属Bradyrhizobium、Dechlorosoma、固氮螺菌属Azospirillum、肠杆菌属Enterobacter、Kosakonia、变形杆菌属Gammaproteobacteria、红长菌属Rubrivivax和Azohydromonas。系统发育树结果表明:3种竹子内生固氮菌明显不同于其他植物。 结论 土壤环境、竹子种类及组织器官均能影响竹子内生固氮菌的结构特征和多样性，定殖于根部的内生固氮菌多样性显著高于叶部。3种竹子内生固氮菌种类与已报道的禾本科Gramineae植物差异显著。图4表4参30
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| 关 键 词： | 竹子 内生固氮菌 PCR-DGGE 聚类分析 Shannon多样性指数 随机克隆 系统发育树 |
| 收稿时间： | 2020-10-10 |
Characteristics and diversity of endophytic diazotrophs in three bamboo species |
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| FENG Gelin,GAO Jing,YAN Shuxian,WANG Jing,LIANG Chenfei,QIN Hua,CHEN Junhui,XU Qiufang.Characteristics and diversity of endophytic diazotrophs in three bamboo species[J].Journal of Zhejiang A&F University,2021,38(6):1203-1212. |
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| Authors: | FENG Gelin GAO Jing YAN Shuxian WANG Jing LIANG Chenfei QIN Hua CHEN Junhui XU Qiufang |
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| Affiliation: | 1.College of Environment and Resources, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China2.State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China |
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| Abstract: | Objective The objective is to investigate the characteristics and biodiversity of endophytic diazotrophs in three bamboo species and in planting soil of root zones. Method Samples of leaves, roots and planting soil from Phyllostachys edulis, Bambusa multiplex and Pleioblastus chino were taken as the objects. The primers of nifH nitrogen fixation gene were designed. PCR-DGGE and random cloning technology were used to construct phylogenetic tree by cluster analysis and Shannon diversity index analysis. Result There were significant differences in community structure of diazotrophs among soils, between roots and leaves of the same bamboo species, between the same organs of different bamboo species, and between the soils of root zones and plants. The similarity of the same bamboo leaf or root was not high. More endophytic diazotrophs were detected in roots than in corresponding leaves. The similarity between diazotrophs in soil and those in roots was higher than that between soil and leaves. Significant differences of diazotrophs among plant species and plant tissues were observed by Shannon diversity index. The Shannon diversity index in leaf (2.73) and soil (2.70) of B. multiplex was higher than that in root (2.53) (P＜0.05), and that in roots of P. chino and Ph. edilus was higher than that in leaf and soil (P＜0.05). 83 strains of diazotrophs were obtained by random cloning and sequencing of plant samples, and only 12 strains were culturable, belonging to Bradyrhizobium, Dechlorosoma, Azospirillum, Enterobacter, Kosakonia, Gammaproteobacteria, Rubrivivax and Azohydromonas. The sequences of phylogenetic tree indicated that the endophytic diazotrophs in the three bamboo species differed greatly from those in other plants. Conclusion The structural characteristics and diversity of diazotrophs in bamboo are affected by soil properties, species and plant tissues. The diversity of endophytic diazotrophs in roots is significantly higher than that in leaves, and the corresponding taxa are distinct from those of Gramineae reported previously. Ch, 4 fig. 4 tab. 30 ref.] |
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