جداسازی ، غربالگری و شناسایی مولکولی باسیلوس های تولید کننده آنزیم اوره آز از اراضی بایر جنوب ‏ایران و بررسی توانایی آنها در تثبیت خاک ‏

نوع مقاله : پژوهشی

نویسندگان

1 دانش‌آموخته دکترای مدیریت حاصلخیزی و زیست‌فناوری خاک، گروه خاک‌شناسی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.

2 استاد گروه خاک‌شناسی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.

3 استاد گروه میکروبیولوژی، دانشکده علوم پایه، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.

4 استادیار گروه زیست‌شناسی، واحد مرودشت، دانشگاه آزاد اسلامی، مرودشت، ایران.

چکیده

اوره آز میکروبی کاربردهای گسترده ای در بیوتکنولوژی، کشاورزی، پزشکی، ساخت و ساز و مهندسی ژئوتکنیک دارد. فناوری ‏رسوب کلسیت به واسطه تحریک میکروبی (‏ MICP‏ ) یک فرآیند اکولوژیکی مبتنی بر فعالیت اوره آز میکروبی است که به ‏تازگی برای تثبیت خاک استفاده شده است. باسیلوس به دلیل قابلیت فروگشت و بقای بالا به عنوان کاندید مناسب برای کاربرد در ‏فرآیند سیمانی شدن زیستی در نظر گرفته شده است. هدف از مطالعه حاضر جداسازی و شناسایی گونه‌های باسیلوس با پتانسیل ‏MICP‏ از اکوسیستم‌های ایران می‏باشد. ‏200 نمونه خاک از اکوسیستم‌های ایران جمع آوری شد و جهت شناسایی باسیلوس‏ها ‏توسط روش های میکروبیولوژیکی فیزیولوژیک و مولکولی از جمله تکثیر ‏PCR‏ و آنالیز توالی ژن های ‏gyrA‏ و ‏‎16S rRNA‎‏ ‏مورد بررسی قرار گرفتند. برای تعیین توانایی تولید سیمان زیستی سویه‏ها، از آزمایشهای رشد در حضوراوره، شوری، ‏pH‏ و ‏دماهای مختلف، ‏SEM، ‏XRD‏ بر روی خاکهای تثبیت شده با سویه ها و آنالیز تونل باد استفاده شد. در مجموع 12 سویه (6%) به ‏عنوان باسیلوس اوره‏آز مثبت شناسایی شد که متعلق به 4 گونه مختلف شامل باسیلوس پارامایکوئیدس 4 سویه(%33/33)، باسیلوس ‏پارالیکنیفورمیس 3 سویه (% 25)، باسیلوس ولزنسیس‏3 سویه(% 25)‏ و باسیلوس پاستوری 2 ایزوله(%66/16) بودند. شرایط بهینه ‏برای ‏MICP‏ توسط سویه ها 30 درجه سانتی گراد، ‏pH9‎‏ و شوری6% است. پس از ‏MICP، نسبت تلفات خاک در سرعت باد 95 ‏کیلومتر در ساعت، 100 برابر کاهش نشان داد. نتایج نشان داد که باسیلوسها، توانایی بالقوه ای برای سازگاری با شرایط سخت ‏محیطی را دارند، همچنین می توان با استفاده از باسیلوسهای تولید کننده ‏MICP‏ در سطح خاک تأثیر به سزایی در کاهش تلفات ‏خاک در اثر فرسایش بادی و افزایش کیفیت آن جهت کشاورزی را دست یافت.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation, screening and molecular identification of bacilli producing urease enzyme from the ‎barren lands of southern Iran and analyzing their ability to stabilize the soil

نویسندگان [English]

  • Somayeh Fazelikia 1
  • Seyed Ali Abtahi 2
  • Mohamad Kargar 3
  • Mojtaba Jafarinia 4
1 Ph.D. graduate, Soil fertility and biotechnology management, Department of Soil Sciences, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.
2 Professor, Department of Soil Sciences, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.‎
3 Professor, Department of Microbiology, Shiraz Branch, Islamic Azad University, Shiraz, Iran.
4 Assistant Prof., Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.
چکیده [English]

Microbial urease has specific applications in biotechnology, agriculture, medicine, construction ‎and geotechnical engineering. MICP technology is an ecological process based on microbial ‎urease activity that has recently been used for soil stabilization. Due to its catabolic ability and ‎high survival, Bacillus is considered as a suitable candidate for use in the biocementation ‎process. The aim of this study was isolating and identifying Bacillus with MICP potential from ‎various ecosystems of Iran.200 environmental samples were collected from the ecosystems of ‎Iran and were analyzed by microbiological and molecular methods including PCR amplification ‎and sequence analysis of gyrA and 16S rRNA genes in order to isolate and identify bacilli. ‎Growth in presence of urea, salinity, pH and different temperatures, SEM, XRD and wind ‎tunnel analysis were used to determine the ability of isolates to produce biocement. A total of ‎‎12 isolates(6%) were identified as urease-positive bacilli belonging to 4 different species, ‎including 4 isolates of Bacillus paramycoides(33.33%), 3 isolates of Bacillus ‎paralicheniformis(25%), and 3 isolates of Bacillus velezensis. isolate(25%) and Bacillus ‎Pasteuri 2 were isolated(16.66%). The optimal conditions for MICP by isolates are 30°C, pH 9 ‎and salinity 6%. After MICP, the ratio of soil loss in the flow rate of 95 km/h showed a 100-‎fold decrease. In conclusion, the results showed that bacilli have the potential ability to adapt to ‎harsh environmental conditions. Also, by using MICP-producing bacilli on the soil surface, we ‎can have a significant effect in reducing soil losses due to erosion.‎

کلیدواژه‌ها [English]

  • biocement
  • bacillus
  • urease
  • soil improvement&lrm

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مدیریت اراضی
دوره 11، شماره 2
دی 1402
صفحه 179-193

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