款冬根际促生细菌的筛选、鉴定及其接种效应

doi:10.11924/j.issn.1000-6850.casb2024-0014

中国农学通报 ›› 2024, Vol. 40 ›› Issue (34): 33-41.doi: 10.11924/j.issn.1000-6850.casb2024-0014

款冬根际促生细菌的筛选、鉴定及其接种效应

张丽慧¹^,²(), 冯丹¹^,², 白变霞¹, 晋婷婷¹, 任嘉红¹()

¹ 长治学院生命科学系，山西长治 046011
² 山西师范大学生命科学学院，太原 030031

收稿日期:2024-01-11 修回日期:2024-10-25 出版日期:2024-12-05 发布日期:2024-12-04
通讯作者:
任嘉红，女，1976年出生，云南文山人，教授，博士，研究方向：植物与微生物互作。通信地址：046011 山西省长治市潞州区太行东街街道长治学院北校区，Tel：0355-2178331，E-mail：renjiahong@163.com。
作者简介:
张丽慧，女，1998年出生，山西大同人，硕士研究生，研究方向：资源微生物。通信地址：046011 山西省长治市潞州区太行东街街道长治学院北校区，E-mail：3171063916@qq.com。
基金资助:
国家自然科学基金“群体感应(QS)介导内生Burkholderia pyrrocinia的生防分子机制”(32071770); 山西省“1331工程”“太行山药用植物生物肥料工程研究中心”(1331KSC); 山西省回国留学人员科研资助项目“款冬褐斑病拮抗合成菌群构建及其活性因子研究”(2023-172)

Screening and Identification of Plant Growth Promoting Bacteria of Tussilago farfara and Its Inoculation Effect

ZHANG Lihui¹^,²(), FENG Dan¹^,², BAI Bianxia¹, JIN Tingting¹, REN Jiahong¹()

¹ Department of Biological Sciences, Changzhi College, Changzhi, Shanxi 046011
² College of Biological Sciences, Shanxi Normal University, Taiyuan 030031

Received:2024-01-11 Revised:2024-10-25 Published:2024-12-05 Online:2024-12-04

摘要/Abstract

摘要：

从款冬根际及根系筛选出PGPR菌株，研究其促生长特性及作用，以期为款冬的丰产栽培提供潜力菌株。采用稀释平板法筛选PGPR菌株，检测其促生特性，通过形态、生理生化指标、BIOLOG及16S rDNA进行鉴定，结合款冬盆栽试验检测其接种效应。结果表明，从款冬根际以及细根中筛选出3株高产IAA菌株KD6B、KD17B和KD26B，鉴定为Enterobacter cloacae subsp. dissolvens、Klebsiella oxytoca和K. areogenes，具解磷、产嗜铁素、耐盐、耐旱及拮抗作用；接种3株菌能显著增加款冬幼苗叶片数、叶面积、根长和根际土壤微生物群落多样性指数，其中KD17B效果最明显，叶片数、叶面积和根长分别增长了36.36%、29.21%、54.07%，平均颜色变化率、Shannon指数和McIntosh指数最大。3株PGPR对款冬有显著促生作用，对其根际土壤微生物群落结构有调控作用，在款冬的丰产栽培中具较高应用价值。

关键词: 款冬, 植物根际促生细菌(PGPR), 筛选, 接种效应, 土壤微生物群落多样性

Abstract:

Growth-promoting characteristics and effects of plant growth promoting bacteria (PGPR) strains isolated from the rhizosphere and root system of Tussilago farfara were studied, in order to provide potential strains for the high-yield cultivation of T. farfara. PGPR strains were obtained by dilution-plating method, and their growth-promoting characteristics were analyzed. These strains were identified by morphological, physiological and biochemical BIOLOG and 16s rDNA sequence. The inoculation effects of these strains on seedlings of T. farfara were examined by a pot-culture experiment. The results showed that three high-yielding IAA strains were selected from the rhizosphere soil and roots of T. farfara, named Enterobacter cloacae subsp. dissolvens (KD6B), Klebsiella oxytoca (KD17B) and K. areogenes (KD26B), which had effects of solubilize phosphorus, produce siderophore, salt tolerance, drought tolerance and antagonism. Three strains significantly increased the number of leaves, leaf area, root length and diversity index of rhizosphere soil microbial community of T. farfara seedlings. Among them, KD17B had the most obvious effect, with the number of leaves, leaf area and root length increasing by 36.36%, 29.21% and 54.07% respectively, and the average color change rate (AWCD), Shannon index and McIntosh index were the largest. Three PGPR strains have significant growth-promoting effects on T. farfara and regulate the structure of rhizosphere soil microbial community, which have high application value in the high-yield cultivation of T. farfara.

Key words: Tussilago farfara, plant growth promoting bacteria (PGPR), screening, inoculation effect, soil microbial community diversity

张丽慧, 冯丹, 白变霞, 晋婷婷, 任嘉红. 款冬根际促生细菌的筛选、鉴定及其接种效应[J]. 中国农学通报, 2024, 40(34): 33-41.

ZHANG Lihui, FENG Dan, BAI Bianxia, JIN Tingting, REN Jiahong. Screening and Identification of Plant Growth Promoting Bacteria of Tussilago farfara and Its Inoculation Effect[J]. Chinese Agricultural Science Bulletin, 2024, 40(34): 33-41.

图/表 16

图1. 3株PGPR菌株的菌落形态

图2. 3株PGPR菌株的IAA显色 A为KD6B，B为KD17B，C为KD26B

图3. 3株PGPR菌株的解磷能力及产嗜铁素能力 A1~A3示解磷能力，A1为KD6B，A2为KD17B，A3为KD26B；B1~B3示产嗜铁素能力，B1为KD6B，B2为KD17B，B3为KD26B

菌株	IAA/(mg/L)	解磷/(mg/L)	嗜铁素能力/%
KD6B	26.95±0.77b	327.0±6.71c	81.62±4.19b
KD17B	27.63±1.46b	568.8±7.32b	30.27±1.65c
KD26B	32.63±0.60a	679.8±6.66a	92.07±3.58a

表1. 3株PGPR菌株的促生长特性

图4. 3株PGPR菌株对NaCl的耐受曲线

图5. 3株PGPR菌株对PEG-6000的耐受曲线

图6. 3株PGPR菌株对A. alternata的平板对抗

图7. 3株菌株的细胞形态

生理生化指标	KD6B	KD17B	KD26B
革兰氏染色	G^-	G^-	G^-
接触酶试验	+	+	+
KOH溶解试验	+	+	+
需氧性试验	好氧	好氧	好氧
油脂水解试验	+	-	-
淀粉水解试验	-	-	-
伏普试验	-	-	+
甲基红试验	-	+	+
吲哚试验	-	-	-
硝酸盐还原试验	-	+	+
纤维素分解试验	-	-	-
明胶水解试验	+	-	-
菌膜形成试验	+	-	-

表2. 菌株的生理生化检测

图8. 3株PGPR菌株的16S rDNA PCR扩增电泳结果泳道1~3分别为KD17B、KD26B、KD6B，M表示Marker

图9. 基于菌株16S rDNA及其同源序列构建的系统发育树

图10. 不同接种处理下幼苗的生长状况 B1为KD6B，B2为KD26B，B3为KD17B

菌株	叶面积/cm²	根长/cm	叶片数量/片
CK	92.62±9.02d	13.5±0.42d	5.5±0.71c
KD6B	159.56±9.67a	17.2±0.28c	6.5±0.71bc
KD26B	141.64±0.51b	19.9±0.71b	8.0±1.41a
KD17B	119.67±3.63c	20.8±0.57a	7.5±0.71ab

表3. 不同接种处理对幼苗的叶面积、根长及叶片数量的影响

图11. 不同接种处理下款冬根际土壤生物群落AWCD变化曲线

图12. 不同接种处理下款冬根际土壤微生物对31种碳源的利用变化特征热图

菌株	Shannon-Wiener指数	Mcintosh指数	Simpson指数
CK	8.09±0.52d	2.90±0.07d	0.94±0.00c
KD6B	9.83±0.51ab	3.18±0.09abc	0.96±0.00ab
KD26B	9.77±0.19ab	3.17±0.05abc	0.96±0.00ab
KD17B	10.13±0.35ab	3.20±0.05abc	0.96±0.00ab

表4. 款冬根际土壤微生物多样性指数

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款冬根际促生细菌的筛选、鉴定及其接种效应

Screening and Identification of Plant Growth Promoting Bacteria of Tussilago farfara and Its Inoculation Effect

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