多粘类芽胞杆菌新菌株X-11及其对番茄和水稻的促生效应

doi:10.13560/j.cnki.biotech.bull.1985.2024-0253

生物技术通报 ›› 2024, Vol. 40 ›› Issue (9): 249-259.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0253

多粘类芽胞杆菌新菌株X-11及其对番茄和水稻的促生效应

刘文志¹^,²(), 贺丹³, 李鹏³, 傅应林¹^,², 张译心¹^,², 温华杰¹^,², 于文清¹^,²^,³()

1.上饶师范学院生命科学学院，上饶 334001
2.上饶师范学院生态环境与绿色发展研究中心赣东北特色植物资源保育与生态修复重点实验室，上饶 334001
3.黑龙江省农垦科学院，哈尔滨 150038

收稿日期:2024-03-16 出版日期:2024-09-26 发布日期:2024-08-01
通讯作者: 于文清，女，博士，教授，研究方向：植物保护；E-mail: wenqingyu09@163.com
作者简介:刘文志，男，研究员，研究方向：生物防治；E-mail: liuwz19801005@sina.com
第一联系人：
贺丹为本文共同第一作者
基金资助:
国家自然科学基金项目(32360698);国家自然科学基金项目(32060633);黑龙江省自然科学基金项目(LH2021C087);高标准农田建设和黑龙江垦区社会公益性事务专项(KJZX202204-11)

Paenibacillus polymyxa New Strain X-11 and Its Growth-promoting Effects on Tomato and Rice

LIU Wen-zhi¹^,²(), HE Dan³, LI Peng³, FU Ying-lin¹^,², ZHANG Yi-xin¹^,², WEN Hua-jie¹^,², YU Wen-qing¹^,²^,³()

1. College of Life Science, Shangrao Normal University, Shangrao 334001
2. Research Center for Eco-Environmental Sciences and Green Development of Shangrao Normal University, Key Laboratory for Regional Plants Conservation and Ecological Restoration of Northeast Jiangxi, Shangrao 334001
3. Heilongjiang Academy of Agricultural Reclamation Sciences, Harbin 150038

Received:2024-03-16 Published:2024-09-26 Online:2024-08-01

摘要/Abstract

摘要：

【目的】从小麦（Triticum aestivum L）根际筛选到了一株细菌菌株（编号X-11），为该菌株在植物促生上应用提供依据。【方法】结合形态特征、生理生化特性、16S rDNA序列及基因组测序对其进行了鉴定，并测定其对番茄（Solanum lycopersicum）和水稻（Oryza sativa）的促生效应。【结果】该菌株菌落圆形、半透明、凸起、黏稠；菌体杆状，形成芽胞，革兰氏阴性；具有固氮，还原硝酸盐，产吲哚、分泌铁载体及分解有机磷的功能。基于16S rDNA序列比对将X-11菌株归类为多粘类芽胞杆菌种群，基于非冗余蛋白数据库的基因比对表明其与最近缘菌株基因相似率55.7%，证明其为一个新菌株。X-11菌液浸种显著增加了番茄发芽率和胚根伸长度，灌根（R）及灌根结合叶面喷施（R+L）显著提高了番茄叶片中过氧化物酶（POD）活性，R+L处理使超氧化物歧化酶（SOD）活性提高了34.58%。水稻1叶1心期施用X-11菌液，根长增加了6.8%；立针期或1叶1心期施用X-11菌液，使秧苗SOD活性分别提高了90.4%和51.8%。【结论】X-11被鉴定为多粘类芽胞杆菌新菌株，是多种功能的植物根际促生菌，在植物促生方面具有开发应用潜力。

关键词: 多粘类芽胞杆菌, 植物根际促生菌, 番茄, 水稻

Abstract:

【Objective】A plant-growth-promoting rhizobacterium strain（named X-11）was screened from rhizosphere soil of wheat（Triticum aestivum L）. Which may provide a basis for the application of the strain in plant growth-promoting.【Method】Strain X-11 was identified by morphological characteristics, physiological and biochemical characteristics, 16S rDNA sequence and genome sequencing, and its effects on the growth and defense enzyme activities of tomato（Solanum lycopersicum）and rice（Oryza sativa）were measured. 【Result】The colonies of strain X-11 are round, swell, neat edge, translucent, and sticky. And the cells are rod-shaped, can form spores, gram negative reaction. And it has the functions of fixing nitrogen, reducing nitrate, producing indole and siderophore, degrading organic phosphorus. Strain X-11 was classified as Paenibacillus polymyxa group based on 16S rDNA sequence, while it was revealed to be a new strain with 55.7% genetic similarity to its closest relative strain through gene comparison in non-redundant database. Tomato（Solanum lycopersicum）seeds soaked with X-11 bacterial suspension increased germination rate and radicle length significantly. Moreover, peroxidase（POD）activity in the leaves significantly increased by X-11 bacterial suspension irrigating roots（R）and irrigating roots combined with spraying leaf surface（R+L）. Meanwhile, the R+L treatment increased superoxide dismutase（SOD）activity by 34.58%. X-11 bacterial suspension increased the seedling root length by 6.8% when applied at one-leaf phase of rice（Oryza sativa）. Additionally, the SOD activity of the seedlings increased by 90.4% and 51.8%, when the X-11 bacterial suspension was applied at vertical phase or one-leaf phase of rice. 【Conclusion】 Strain X-11 was identified as a new strain of P. polymyxa, which is the effective plant-growth promoting rhizobacterium with multiple functions and has potential for development and application in plant growth promotion.

Key words: Paenibacillus polymyxa, plant-growth promoting rhizobacterium, tomato, rice

刘文志, 贺丹, 李鹏, 傅应林, 张译心, 温华杰, 于文清. 多粘类芽胞杆菌新菌株X-11及其对番茄和水稻的促生效应[J]. 生物技术通报, 2024, 40(9): 249-259.

LIU Wen-zhi, HE Dan, LI Peng, FU Ying-lin, ZHANG Yi-xin, WEN Hua-jie, YU Wen-qing. Paenibacillus polymyxa New Strain X-11 and Its Growth-promoting Effects on Tomato and Rice[J]. Biotechnology Bulletin, 2024, 40(9): 249-259.

图/表 8

图1 多粘类芽胞杆菌X-11平板菌落形态和显微镜下细胞形态 A和B、C和D、E和F、G和H分别为X-11菌株在阿须贝无氮培养基、硅酸盐细菌培养基、马铃薯葡萄糖培养基、马铃薯蔗糖培养基平板上的菌落形态，I和J分别为X-11在复合培养基中培养24和48 h后细胞在显微镜下的形态

Fig. 1 Morphology of P. polymyxa X-11 colony on plate and cell morphology under microscope A and B, C and D, E and F, G and H, refer to the colony morphology of X-11 strain on the Ashby’s nitrogen-free medium, silicate bacteria medium, potato dextrose medium, potato sucrose medium agar plate, respectively. I and J refer to the morphology of X-11 cell under the microscope after 24-h and 48-h cultivation in complex broth

表1 多粘类芽胞杆菌X-11生理生化特性及糖酵解试验结果

Table 1 Results of physiological and biochemical characteristics and glycolysis test of P. polymyxa X-11

生理生化反应 Physiological and biochemical reaction	测定结果 Determination result	糖酵解反应 Glycolysis reaction	测定结果 Determination result
生理生化反应 Physiological and biochemical reaction	测定结果 Determination result	糖酵解反应 Glycolysis reaction	产酸Acid production	产气Aerogenesis
柠檬酸盐利用 Citrate utilization	＋	葡萄糖Glucose	＋	＋
酪素水解 Casein hydrolysis	＋	蔗糖 Sucrose	＋	＋
淀粉水解 Starch hydrolysis	＋	甘露醇 Mannitol	＋	＋
油脂水解 Lipid hydrolysis	＋	乳糖 Lactose	＋	＋
明胶水解 Gelatin hydrolysis	＋	山梨醇 Sorbitol	＋	＋
V-P反应 The V-P reaction	＋	阿拉伯糖 Arabinose	＋	＋
硝酸盐还原 Nitrate reduction	＋	水杨苷 Salicin	＋	＋
甲基红变色 Methyl red color	-	昆布多糖Laminarin	＋	＋
吲哚试验 Indole test	＋	木聚糖Xylan	＋	-
氧化酶活性 Oxidase activity	＋	木糖 Xylose	-	＋
接触酶活性 Contactase activity	＋	肌醇 Inositol	-	-
酪氨酸水解 Tyrosine hydrolysis	＋	侧金盏醇 Lateral calendula alcohol	-	-
石蕊牛奶变色 Litmus milk discoloration	＋	甘油 Glycerin	-	-
过氧化氢酶活性 Catalase activity	-

图2 基于16S rDNA的多粘类芽胞杆菌X-11系统发育树（A）与基于非冗余蛋白数据库的X-11与近缘菌株基因分布饼图（B）系统发育树中（A）菌株前的编码为16S rDNA在NCBI基因库中的登录码，分支处的数字为相邻2个分支的相似性（%）。饼图（B）中数字代表在非冗余蛋白数据库注释上的X-11基因中与各菌株匹配上的数目

Fig. 2 Evolutionary tree of P. polymyxa X-11 based on 16S rDNA（A）, and pie diagram of gene distribution of P. polymyxa X-11 and relative strains based on non-redundant protein sequence database（B） In the evolutionary tree（A）, the 16S rDNA entry code of the NCBI GenBank was followed by strain name, the number at the branch is the identification rate of the two adjacent strains. The numbers in the pie diagram（B）indicate the number of matches to each strain in the X-11 genes annotated in the non-redundant protein database

图3 不同浓度多粘类芽胞杆菌X-11菌悬液催芽处理下番茄种子发芽率 A：‘粉冠’品种；B：‘金棚1号’品种。不同小写字母代表处理间差异显著（P<0.05）。下同

Fig. 3 Germination rate of tomato seeds under different concentrations of P. polymyxa X-11 suspension A: ‘Fenguan’ variety. B: ‘Jinpeng No. 1’ variety. Different lower letters on the column indicate difference among treatments are significantly（P<0.05）. The same below

图4 多粘类芽胞杆菌X-11不同浓度处理下番茄胚根长度 A-I为番茄‘粉冠’品种，J-R为番茄‘金棚1号’品种；B-I和K-R分别为X-11菌悬液稀释1、10、50、100、200、300和1 000倍及空白对照（0）的种子胚根长度

Fig. 4 Radicle length of tomato treated with different concentrations of P. polymyxa X-11 A-I are the ‘Fenguan’ variety, J-R are the ‘Jinpeng No. 1’ variety; B-I and K-R are the seed radicle length of 1, 10, 50, 100, 200, 300, and 1 000 times dilution of X-11 bacterial suspension, and blank control（0）, respectively

图5 多粘类芽胞杆菌X-11及多菌灵不同处理方式下番茄叶片防御酶活性 R、L、R+L及Con分别代表叶面喷施、灌根、灌根结合叶面喷施及对照；CA代表多菌灵，X-11代表多粘类芽胞杆菌X-11。柱上不同小写字母代表处理方式间差异显著，不同大写字母代表同一处理方式下CA与X-11之间差异显著（P<0.05）

Fig. 5 Defense enzyme activities of tomato leaves under different treating methods of P. polymyxa X-11 and carbendazim The horizontal coordinate R, L, R+L and Con refers to foliar spraying, root irrigation, root irrigation combined foliar spraying, and control respectively. Legend CA and X-11 refers to carbendazim and P. polymyxa X-11, respectively. Different lowercase letters on the column indicate difference among treatment methods is significantly, and different uppercase letters indicate the difference between CA and X-11 under the same treatment is significant (P<0.05)

图6 多粘类芽胞杆菌X-11菌液不同施用时期的水稻株高（A）及根长（B） V、1、2和Con分别代表立针期、1叶1心期、2叶1心期和对照。下同

Fig. 6 Rice shoot length（A）and root length（B）at different application periods of Paenibacillus polymyxa X-11 The horizontal coordinates V, 1, 2 and Con represent the vertical needle phase, the one-leaf phase, the two-leaves phase and control, respectively. The same below

图7 多粘类芽胞杆菌X-11菌液不同施用时期的水稻防御酶活性

Fig. 7 Rice defense enzyme activities at different application periods of P. polymyxa X-11 bacterial suspension

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多粘类芽胞杆菌新菌株X-11及其对番茄和水稻的促生效应

Paenibacillus polymyxa New Strain X-11 and Its Growth-promoting Effects on Tomato and Rice

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