大豆根瘤内生细菌的分离鉴定及其对大豆植株的促生效应

doi:10.3969/j.issn.1004-1524.20221120

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (7): 1532-1541.DOI: 10.3969/j.issn.1004-1524.20221120

大豆根瘤内生细菌的分离鉴定及其对大豆植株的促生效应

孙秀娟(), 徐伟慧, 王志刚()

齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔 161006

收稿日期:2022-08-02 出版日期:2023-07-25 发布日期:2023-08-17
作者简介:孙秀娟(1998—),女,黑龙江鸡西人,硕士研究生,主要从事根际微生物研究。E-mail: 1585289485@qq.com
通讯作者: *王志刚,E-mail: wangzhigang@qqhru.edu.cn
基金资助:
黑龙江省重点研发计划项目(GA21B007);黑龙江省重点研发计划项目(GZ20210067)

Isolation and identification of endophytic bacteria from soybean nodule and their effects on soybean plant

SUN Xiujuan(), XU Weihui, WANG Zhigang()

College of Life Science and Agriculture and Forestry, Qiqihar University, Qiqihar 161006, Heilongjiang, China

Received:2022-08-02 Online:2023-07-25 Published:2023-08-17
Contact: WANG Zhigang

摘要/Abstract

摘要：

为探究大豆根瘤内生细菌对大豆的促生效应,以大豆根瘤为实验材料,分离筛选根瘤内生细菌,测定其分泌植物激素能力及其对大豆株高、茎粗、干重、鲜重和根系生长方面的促生能力。结果表明:共筛选出17株根瘤内生细菌,经生理生化和16S rDNA鉴定,有10株为芽孢杆菌属(Bacillus),3株为巨大普里斯特氏菌属(Priestia),其余4株分别为肠杆菌属(Enterobacter)、克雷伯氏菌属(Klebsiella)、不动杆菌属(Acinetobacter)和农杆菌属(Agrobacterium)。通过溶血试验和16S rDNA鉴定筛选出10株促生能力强的菌株,其中,Klebsiella S8的吲哚乙酸(IAA)分泌量最高,为147.55 mg·L^-1;赤霉素(GAs)分泌量最高的为Agrobacterium S10,分泌量达39.34 mg·L^-1;Bacillus S3的铁载体活性最高,为70.57%;10株菌株处理的大豆株高、根长、干鲜重和茎粗均显著高于对照。综上所述,筛选的大豆根瘤内生细菌,对大豆具有较强的促进作用。

关键词: 根瘤, 内生细菌, 大豆, 溶血试验

Abstract:

To study the growth-promoting effect of endophytic bacteria in soybean root nodules on soybean growth, soybean root nodules were used as experimental materials to isolate and screen nodule endophytic bacteria, and their ability to secrete plant hormones and their promotion effect on soybean plant height, stem diameter, dry weight, fresh weight, and root growth were measured. The results showed that a total of 17 endophytic bacteria were screened out from root nodules. After physiological and biochemical analysis and 16S rDNA identification, 10 strains belonged to Bacillus spp., 3 strains belonged to Priestia spp, and the remaining 4 strains belonged to Enterobacteriaceae, Klebsiella, Acinetobacter, and Agrobacterium. Through hemolysis test and 16S rDNA identification, 10 strains with strong growth-promoting ability were selected. Among them, the IAA secretion of Klebsiella S8 was highest at 147.55 mg·L^-1; the gibberellins of Agrobacterium S10 had the highest secretion at 39.34 mg·L^-1; the siderophores activity of Bacillus S3 was the highest at 70.57%. The plant height, root length, dry fresh weight and stem diameter of soybean treated by these 10 strains were significantly higher than those of the control. In conclusion, the endophytic bacteria screened in this experiment had strong growth-promoting effect on soybean growth.

Key words: root nodule, endophytic bacterium, soybean, hemolysis test

中图分类号:

孙秀娟, 徐伟慧, 王志刚. 大豆根瘤内生细菌的分离鉴定及其对大豆植株的促生效应[J]. 浙江农业学报, 2023, 35(7): 1532-1541.

SUN Xiujuan, XU Weihui, WANG Zhigang. Isolation and identification of endophytic bacteria from soybean nodule and their effects on soybean plant[J]. Acta Agriculturae Zhejiangensis, 2023, 35(7): 1532-1541.

图/表 5

图1 基于16S rDNA序列的内生细菌进化树

Fig.1 Phylogenetic tree of endophytic bacteria based on 16S rDNA sequences

表1 内生细菌的部分生理生化特性

Table 1 Physiological and biochemical characteristics of endophytic bacteria

菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
S2	+	-	+	+
S3	+	-	+	+
S4	-	+	-	+
S5	+	-	+	-
S6	+	-	-	-
S7	-	+	+	+
S8	-	+	+	+
S9	-	+	-	+
S10	-	+	+	-

图2 供试菌株分泌IAA、GAs和铁载体的能力柱上无相同小写字母表示差异显著(P<0.05)。下同。

Fig.2 Ability of the tested strains to secrete IAA, gibberellin and siderophores Data on the bars marked without the same lowercase letter indicated significant differences at P<0.05. The same as below.

图3 内生细菌对大豆种子的促生作用幼苗生长图3个为1个处理,从左至右依次是CK、稀释10倍、20倍和30倍。

Fig.3 Growth promoting effect of endophytic bacteria on soybean seed The three seedling growth plots are one treatment, from left to right are CK, diluted 10 times, 20 times and 30 times.

图4 菌株对大豆植株促生影响与综合能力评价 F,综合能力评价。

Fig.4 Effect of endophytic bacteria on growth promotion of soybean plants and comprehensive ability evaluation F, Evaluation of comprehensive ability.

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大豆根瘤内生细菌的分离鉴定及其对大豆植株的促生效应

Isolation and identification of endophytic bacteria from soybean nodule and their effects on soybean plant

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菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
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菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
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[9]	孟娜, 薛辉, 魏明, 魏胜华. 氯通道抑制剂缓解栽培大豆盐伤害的离子特征[J]. 浙江农业学报, 2022, 34(10): 2095-2104.
[10]	杨昕霞, 张斌. 大豆LAZ1基因家族鉴定与GmLAZ1-9基因的功能研究[J]. 浙江农业学报, 2021, 33(4): 586-594.
[11]	张伟梅, 张古文, 冯志娟, 刘娜, 王斌, 卜远鹏. 菜用大豆籽粒中蔗糖的遗传与调控机制研究进展[J]. 浙江农业学报, 2021, 33(12): 2446-2456.
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菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
S2	+	-	+	+
S3	+	-	+	+
S4	-	+	-	+
S5	+	-	+	-
S6	+	-	-	-
S7	-	+	+	+
S8	-	+	+	+
S9	-	+	-	+
S10	-	+	+	-

菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
S2	+	-	+	+
S3	+	-	+	+
S4	-	+	-	+
S5	+	-	+	-
S6	+	-	-	-
S7	-	+	+	+
S8	-	+	+	+
S9	-	+	-	+
S10	-	+	+	-

菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
S2	+	-	+	+
S3	+	-	+	+
S4	-	+	-	+
S5	+	-	+	-
S6	+	-	-	-
S7	-	+	+	+
S8	-	+	+	+
S9	-	+	-	+
S10	-	+	+	-

菌株编号 Strain No.	淀粉水解试验 Starch hydrolysis test	油脂水解试验 Oil hydrolysis test	乳糖发酵试验 Lactose fermentation test	葡萄糖发酵试验 Glucose fermentation test
S1	+	-	+	+
S2	+	-	+	+
S3	+	-	+	+
S4	-	+	-	+
S5	+	-	+	-
S6	+	-	-	-
S7	-	+	+	+
S8	-	+	+	+
S9	-	+	-	+
S10	-	+	+	-