水稻内生固氮菌分离鉴定、生物特性及其对稻苗镉吸收的影响

doi:10.3864/j.issn.0578-1752.2016.19.008

Abstract

Abstract: 【Objective】The objective of this study is to isolate, identify and analyze phylogenetics of endophytic diazotrophs from rice planted in Hunan province, test the biological characteristics of the isolates, and to explore the influences of diazotroph inoculation on rice seedlings Cd accumulation. 【Method】Surface sterilization and low nitrogen medium were used to isolate endophytic diazotrophs. nifH detection was conducted based on PCR amplification to confirm the isolates as nitrogen-fixing bacteria. 16S rRNA was amplified with PCR, blasted in EzTaxon after sequencing, and analyzed with Clustalx-MEGA to make phylogenetic tree. Greenhouse trials were conducted to investigate the influence of diazotroph inoculation on rice seedling Cd accumulation. 【Result】Nineteen endophytic diazotrophs were isolated from root, stem and leaf of 8 rice samples. These 19 strains phylogenetically belong to 4 genus 13 species of Bacillus aryabhattai, B. cereus, B. idriensis, B. sindicus, B. licheniformis, B. megaterium, B. methylotrophicus, B. subtilis, B. tequilensis, Brevibacterium. halotolerans, Fictibacillus phosphorivorans, Paenibacillus barcinonensis, P. lautus. Biological tests showed that about 1/3 of the 19 strains produce protease and cellulose, grow well at 48℃, form spores well with percentage 60%-90%, produce alkali with final pH 8.5-9.0. About 1/6 of the 19 strains are antagonistic against plant pathogenic Rhizoctonia solani ACCC36246, Fusarium graminearum ACCC36249 and Fusarium sporotrichioides ACCC37402 with rate of 42%-55%. About 2/3 of the 19 strains showed sensitive to antibiotics and resistant to fungicide. Four representative strains of the 19 could utilize 7 of the 78 carbon sources, sodium lactate, sucrose, dextrose, glycerol, malic acid, alanine and glucuronic acid amide. Greenhouse trials showed that 6 of the 19 strains promoted rice seedling Cd absorption with increase of 6.41%-38.45%, and other 13 strains decreased rice seedling Cd absorption with 2.06%-34.46% compared with control.【Conclusion】Nineteen endophytic diazotrophs were isolated from rice planted in Hunan. These 19 strains phylogenetically belong to 4 genus 13 species of Bacillus, Brevibacterium, Fictibacillus and Paenibacillus. Partial strains produce protease and cellulose, grow well at 48℃, form spores well, antagonistic against plant pathogenic Rhizoctonia solani, Fusarium graminearum and Fusarium sporotrichioides, have good prospects of application. Inoculation of diazotroph can significantly affect rice seedling Cd absorption. The results suggest that application of microbial method to control paddy Cd is a very worthwhile pathway.

Key words: rice, endophytic diazotrophs, biological property, Cd

YUAN Mei, TAN Shi-juan, SUN Jian-guang. Isolation and Biological Properties of Endophytic Diazotrophs from Rice and Their Influences on Rice Seedling Cd Accumulation[J].Scientia Agricultura Sinica, 2016, 49(19): 3754-3768.

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