جداسازی و شناسایی باکتری‌های فیلوسفر ذرت و تعیین ویژگی های محرک رشدی گیاه آنها

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه علوم خاک، دانشگاه شیراز

2 استادیار ، گروه علوم خاک، دانشگاه شیراز،

3 استاد مؤسسه تحقیقات خاک و آب، سازمان تحقیقات، آموزش و ترویج کشاورزی

4 دانشیار،گروه علوم خاک، دانشگاه شیراز،

5 استاد، گروه علوم خاک، دانشگاه شیراز

6 استاد، گروه گیاهپزشکی، دانشگاه شیراز

چکیده

سابقه و هدف: باکتری‌های افزایش دهنده رشد گیاه ساکن در فیلوسفر گیاهان مختلف ابزاری برای استفاده کارآمدتر از باکتری‌ها به عنوان منبعی برای تولید کودهای زیستی را فراهم می‌کند. استفاده از فن‌آوری‌های مولکولی منجر به پژوهش عمیق در مورد ریزجانداران فیلوسفری می‌شود و ممکن است راه‌های جدیدی را برای توسعه و اعتباردهی عملیات کشاورزی مدرن فراهم نموده و سبب افزایش چشمگیر بهره‌وری شود. هدف از این پژوهش بررسی ویژگی‌های محرک رشد باکتری‌های جدا شده از فیلوسفر ذرت (Zea mays L.) رشد یافته در مناطق مختلف جغرافیایی کشور (ایران) و شناسایی مولکولی آنها با روش توالی‌یابی ژن 16Sr RNA بود.
مواد و روش‌ها: در شهریور سال 1395، 116 نمونه‌ برگ سالم بالغ از مزارع ذرت واقع در 6 استان ایران (البرز، تهران، خراسان رضوی، خوزستان، فارس و قزوین) جمع‌آوری و سپس باکتری‌های فیلوسفری از آنها جداسازی شدند. ویژگی‌های محرک رشد باکتری‌ها از جمله، تولید ایندول استیک اسید (Indole-3 Acetic acid, IAA)، انحلال فسفات آلی نامحلول، تولید سیدروفور، تولید پلی‌ساکاریدهای برون‌سلولی (Exopolysaccharides, EPS) و نیز فعالیت آنزیم نیتروژناز با استفاده از روش‌های استاندارد اندازه گیری شدند. سویه‌های برتر بر اساس روش توالی‌یابی ژن 16Sr RNA در حد گونه شناسایی شدند.
یافته‌ها: 10 سویه از 242 باکتری‌ جداسازی شده از فیلوسفر ذرت براساس ویژگی‌های محرک رشد آنها انتخاب شدند. همه سویه‌های انتخابی دارای فعالیت آنزیم نیتروژناز، انحلال فسفات آلی و قابلیت تولید IAA و EPS بودند. بیشترین فعالیت آنزیم نیتروژناز (31/70nM ml-1 h-1 ) و انحلال فسفات آلی (05/75 g l-1) در سویه THE15 مشاهده شد، اگرچه کمترین مقدار این ویژگی‌ها به ترتیب متعلق به سویه‌های KHO57 (55/5 nM ml-1 h-1) و KHO51 (90/15 g l-1) بودند. سویه‌های QAZ26 و TEH15 دارای بیشترین مقدار IAA (به ترتیب 56/31 و 00/30 μg ml-1) بودند و سویه ALB32 کمترین مقدار IAA (50/4 μg ml-1) را تولید کرد. تولید EPS به‌وسیله سویه‌ها بین 805/1-120/0 g l-1 متغیر بود. در میان سویه‌های بررسی شده، تنها KUU32، KHU53، ALB32 و QAZ26 توانستند سیدروفور تولید کنند. توالی‌یابی 16S rRNA نشان داد که 10 سویه متعلق به جنس‌های Bacillus، Microbacterium، Pseudarthrobacter، Pseudomonas، Stenotrophomonas، Enterobacter و Kocuria هستند. Bacillus با 4 گونه مختلف (شامل Bacillus subtilis، Bacillus megaterium، Bacillus paralicheniformis و Bacillus pumilus) غالب‌ترین جنس بود.
نتیجه‌گیری: بر اساس نتایج این پژوهش باسیلوس‌ غالب‌ترین جنس باکتری‌های افزایش دهنده رشد در فیلوسفر ذرت بود. از میان باکتری‌های جداسازی شده، TEH15 Enterobacter hormaechei توانایی بالایی در فعالیت آنزیم نیتروژناز، تولید ایندول استیک اسید و انحلال فسفات آلی داشت. بنابراین این سویه شایسته پژوهش بیشتر در بررسی‌های میدانی (مطالعات گلخانه‌ای و مزرعه‌ای) کشت ذرت و گیاهان هم‌خانواده ذرت است.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation and identification of maize phyllosphere bacteria and determination of their plant growth promoting traits

نویسندگان [English]

  • Vahid Alah Jahandideh Mahjen Abadi 1
  • mozhgan Sepehri 2
  • Hadi Asadi Rahmani 3
  • Mehdi Zarei 4
  • Abdolmajid Ronaghi 5
  • Seyed Mohsen Taghavi 6
1 Dept. of Soil Sci., College of Agric., Shiraz Univ.
2 Soil Science Department, College of Agriculture, Shiraz University
3 Soil and Water Research Institute, Agriculture Research, Education and Extension Organization
4 Department of Soil Science, Shiraz University
5 Shiraz university
6 Department of Plant pathology, Shiraz University
چکیده [English]

Background and Objectives: The plant growth promoting bacteria (PGPB) inhabiting the phyllosphere of different plants provides a tool for more efficient use of bacteria as a source of biofertilizers. The use of molecular technologies leads to in-depth research of the phyllosphere microorganisms and may provide new avenues for the development and validation of modern agricultural operations and significantly increase productivity. The aim of this research was to investigate the plant growth promoting (PGP) traits of bacteria isolated from maize (Zea mays L.) phyllosphere cultivated in different geographical zones of country (Iran) and their molecular identification by 16Sr RNA sequencing.

Materials and Methods: In September (2016), 116 samples of healthy mature leaves were collected from the maize fields located in 6 provinces of Iran (Fars, Alborz, Qazvin, Khuzestan, Tehran and Khorasan) and then, phyllosphere bacteria were isolated from them. Growth promoting traits of bacteria including Indole-3 Acetic acid (IAA) production, organic phosphate dissolution, siderophore production, exopolysaccharides (EPS) production and also nitrogenase enzyme activity were measured using standard methods. Superior strains were identified based on 16S rRNA sequencing method.

Results: The 10 from 242 bacterial strains isolated from maize phyllosphere were selected based on their growth promoting traits. All strains selected demonstrated nitrogenase activity, organic phosphate dissolution and IAA and EPS production capability. The highest nitrogenase activity (70.31 nM ml-1 h-1) and organic phosphate dissolution (75.05 g l-1) were observed in THE15 strain, while the lowest amount of these properties belonged to KHO57 (5.55 nM ml-1 h-1) and KHO51 (15.90 g l-1) strains, respectively. The QAZ26 and TEH15 strains had the highest amount of IAA production (31.56 and 30.00 µg ml-1, respectively) and the ALB32 strain produced the lowest amount of IAA (4.50 µg ml-1). The EPS production by strains ranged from 0.120 to 1.805 g l-1. Among the strains investigated, only KUU32, KHU53, ALB32 and QAZ26 were able to produce siderophore. 16S rRNA sequencing revealed that the 10 strains selected belong to Bacillus, Microbacterium, Pseudarthrobacter, Pseudomonas, Stenotrophomonas, Enterobacter and Kocuria genera. The Bacillus with 4 different species (including Bacillus subtilis, Bacillus megaterium, Bacillus paralicheniformis and Bacillus pumilus) was the most dominant genus.

Conclusion: Based on the results of this study, Bacillus was the most dominant genus of plant growth promoting bacteria in the maize phyllosphere. Among the isolated bacteria, Enterobacter hormaechei TEH15 was highly capable of nitrogenase enzyme activity, IAA and organic phosphate solubility. Therefore, this strain deserves further research in greenhouse and field studies of maize cultivation and maize family plants.

کلیدواژه‌ها [English]

  • Phyllosphere microbiome
  • Plant growth promoting traits
  • 16S rRNA
  • Enterobacter hormaechei TEH15

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مجله مدیریت خاک و تولید پایدار
دوره 10، شماره 2
شهریور 1399
صفحه 47-67

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