促进水稻铁素吸收的野生稻内生细菌优良菌株的筛选与鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2024-0178

摘要/Abstract

摘要：

【目的】为筛选促进水稻高效吸收铁元素的嗜铁内生细菌，探究野生稻中不同种属的内生细菌的产铁载体能力和促进水稻铁素吸收效果。【方法】对前期从4种野生稻（Oryza longistaminata、O. rufipogon、O. officinalis和O. minuta）中分离获得的198株内生细菌进行产铁载体定量测定分析，从128株具有产铁载体能力的内生菌株中筛选获得82株高产铁载体内生菌株；16S rRNA基因序列分析显示82个菌株属于18个种属；结合其产铁载体能力，选择18株产铁载体候选菌株进行促进水稻铁吸收效果鉴定。【结果】不同种属的功能菌株促进铁吸收效果有差异，其中（类）芽孢杆菌属、假单胞菌属和考氏科萨克氏菌属促进铁吸收效果明显，根据促生试验从18株菌株中筛选出了5株最优的高效产铁载体菌株，并进一步验证了其可以部分恢复水稻铁吸收缺陷突变体（osblhl156）的缺铁黄化表型。【结论】明确了植物互作微生物所产的铁载体可以被水稻利用，丰富了水稻铁吸收相关微生物资源，为水稻-微生物互作促进水稻铁吸收提供菌种资源和理论依据。

关键词: 水稻, 内生细菌, 铁载体, 植物-微生物互作, 铁吸收

Abstract:

【Objective】To identify siderophilic endophytic bacteria that efficiently promote iron absorption in rice, and to investigate the siderophore-producing capacity and iron absorption promoting effect of different genera of endophytic bacteria from four wild rice.【Method】This study analyzed the siderophore contents of 198 endophytic bacteria isolated from four types of wild rice including Oryza longistaminata, O. rufipogon, O. officinalis, and O. minuta. The 82 strains with highly efficient siderophore-producing ability were screened from 128 strains with siderophore-producing ability. Analysis of the 16S rRNA gene sequences revealed that these 82 strains belonged to 18 different genera or species. Based on their siderophore production abilities, 18 candidate strains were selected to assess their abilities of promoting iron absorption in rice.【Result】The results demonstrated that different genera had varying effects on promoting rice iron absorption, with Bacillus（class）, Pseudomonas, and Saccharomycetes showing significant effects. Five high-efficient siderophore producing strains were selected from 18 strains and further confirmed that endophytic bacteria partially restored the iron deficiency phenotype of a rice iron-absorption-deficient mutant（osbhlh156）.【Conclusion】This study elucidates that siderophore produced by plant-interacting microorganisms can be utilized by rice, thereby enriching the microbial resources associated with iron absorption in rice. Additionally, it provides valuable bacterial resources and a theoretical basis for promoting iron absorption in rice through rice-microbial interactions.

Key words: rice, endophytic bacteria, siderophores, plant - microbial interaction, iron absorption

李庆懋, 彭聪归, 齐笑含, 刘兴蕾, 李臻园, 李沁妍, 黄立钰. 促进水稻铁素吸收的野生稻内生细菌优良菌株的筛选与鉴定[J]. 生物技术通报, 2024, 40(8): 255-263.

LI Qing-mao, PENG Cong-gui, QI Xiao-han, LIU Xing-lei, LI Zhen-yuan, LI Qin-yan, HUANG Li-yu. Screening and Identification of Excellent Strains of Endophytic Bacteria Promoting Rice Iron Absorption from Wild Rice[J]. Biotechnology Bulletin, 2024, 40(8): 255-263.

图/表 5

图1 内生菌产铁载体能力分析 A：四种野生稻中产铁载体最强的产铁载体功能菌株的平板示意图；B：四种野生稻分离的产铁载体菌株mp值；C：128株内生细菌产铁载体能力定量分析PCA图。OL表示长雄野生稻，OR表示普通野生稻，OO表示药用野生稻，OM表示小粒野生稻

Fig. 1 Siderophore-producing ability analysis of isolated endophytic bacteria A: Plate schematic diagram of strains with the strongest siderophore-producing function from four wild rices. B: mp values of siderophore contents of endophytic bacteria among four wild rices. C: PCA diagram of siderophore contents produced by 128 endophytic bacteria. OL refers to Oryza longistaminata, OR to O. rufipogon, OO to O. officinalis, and OM to O. minuta

图2 82株内生细菌多样性分类（A）和18株不同种属候选菌株的产铁载体能力（B）

Fig. 2 Diversity classification of 82 endophytic bacteria (A)and siderophore-producing ability of 18 candidate strains of different genera/species (B)

图3 不同菌株浸种处理后水稻表型鉴定不同内生菌处理后苗期形态图（A）；苗期株高（B）、根长（C）、叶绿素含量（D）和铁含量（E）统计分析。比例尺10 cm；标有*表示与对照相比达到差异显著（P < 0.05）

Fig. 3 Phenotyping of rice treated by different endophytic bacteria with siderophore-producing ability Morphological phenotype（A）and statistical analysis of plant height（B）, root length（C）, chlorophyll（D）, and iron content（E）after treated with different endophytic bacteria. Scale bar: 10 cm; * indicates significant difference compared with the control（P < 0.05）

表1 18株产铁载体菌株对ZH11株高、根长、叶绿素和铁含量的促进效果

Table 1 Effect of 18 siderophore-producing strains on the plant height, root length, chlorophyll and iron content of ZH11

接种菌株 Inoculated strain	生长和生理指标Growth and physiological indicators
接种菌株 Inoculated strain	株高 Plant height/cm	根长 Root length/cm	叶绿素含量（SPAD）Chlorophyll content （SPAD）	铁含量 Total iron content/（mg·kg^-1）
OML3-3	-	+	+	+
OMR1-7	+	+	+	+
OOS1-3	-	+	+	+
OOS2-3	-	-	+	+
ORL1-5	-	-	+	-
OML3-6	+	+	+	+
ORL1-9	-	-	+	+
ORR1-18	+	-	+	-
OLL1-12	-	-	-	-
OML2-8	-	-	+	-
ORR3-11	-	-	-	+
OOR2-3	-	-	-	-
OMS2-7	-	+	-	+
OMR2-7	+	-	+	+
OOL1-5	+	-	-	+
ORR3-14	-	-	-	+
OLL1-10	-	+	-	+
OLL2-1	+	+	-	-

图4 高效产铁载体菌株部分恢复osbhlh156突变体缺铁失绿表型产铁载体菌株处理前后植株形态表型（A）、铁含量（B）和叶绿素含量（C）。比例尺5 cm；标有*表示与对照相比达到差异显著（P < 0.05）

Fig. 4 Phenotype of osbhlh156 mutants recovered by highly efficient candidate strains Morphological phenotype（A）and iron content（B）and chlorophyll content（C）of osbhlh156 plants before and after treated with endophytic bacteria. Scale bar: 5 cm. * indicates significant difference compared with the control（P < 0.05）

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