جداسازی باکتری‌های متحمل به مس از یک خاک آلوده، شناسایی و بررسی خصوصیات محرک رشدی آنها

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد دانشکده کشاورزی دانشگاه ولی‌عصر (عج) رفسنجان

2 استادیار گروه علوم خاک دانشکده کشاورزی دانشگاه ولی‌عصر (عج) رفسنجان

3 دانشیار گروه علوم خاک دانشکده کشاورزی دانشگاه ولی‌عصر (عج) رفسنجان

چکیده

تلقیح گیاهان با باکتری­های ریزوسفری محرک رشد موثر و مقاوم به فلزات سنگین در خاک­های آلوده به منظور افزایش کارایی گیاه­پالایی و استخراج فلز سنگین از خاک توسط گیاه امری ضروری می­باشد. در این جهت این پژوهش به منظور جداسازی، شناسایی و صفات محرک رشدی باکتری­های مقاوم مس از یک خاک آلوده به مس انجام شد. ابتدا شش نمونه خاک ریزوسفری گیاهان بومی منطقه خاتون­آباد سرچشمه استان کرمان جمع­آوری شد و نهایتا شش جدایه باکتری از نمونه­های خاک ریزوسفری جمع­آوری و خالص­سازی شد. نتایج شناسی فنوتیپی، بیوشیمیایی و ژنتیکی نشان داد که پنج جدایه از جنس سودوموناس و یک جدایه از جنس باسیلوس بودند. سپس حداقل غلظت بازدارنده[1] مس، میزان تولید سیدروفور، تولید اکسین، سیانید هیدروژن، توانایی حل فسفات­های معدنی، در محیط جامد و مایع، اندازه‌گیری توان حل­کنندگی ترکیبات کم محلول روی در محیط جامد و مایع و تولید -ACC دآمیناز باکتری­ها بررسی شد. نتایج نشان داد تمام جدایه­ها نسبت به غلظت­های مختلف مس از خود مقاومت نشان دادند. بیشترین مقاومت نسبت به غلظت بازدارنده مس مربوط به سویه­های K4 و K5 در غلظت 400 میلی­گرم بر لیتر مس بود. همچنین در رابطه با تولید سیدروفور، بجز سویه­های K6 که توانایی تولید سیدروفور را در محیط CAS نداشت، پنج سویه دیگر توانایی تولید سیدروفور را در این محیط دارا بودند و سویه ی K5 بیشترین میزان تولید سیدروفور با نسبت هاله به کلنی 56/1 را به خود اختصاص داد. در بین جدایه­ها، دو جدایه K4 و K6 قادر به تولید اکسین نبودند. تولید اکسین توسط جدایه K1 تفاوت معنی­داری با دیگر جدایه­های مورد بررسی داشته و با مقدار 07/2 میلی­گرم در لیتر بیشترین میزان تولید اکسین در بین جدایه­ها را دارا بود. نتایج همچنین نشان داد که از بین جدایه­ها، تنها جدایه K4 توانایی تولید سیانید هیدروژن را با درجه چهار (نسبتا زیاد) دارا بوده و دیگر جدایه ها با اختصاص درجه یک مبنی بر عدم تولید سیانید هیدروژن گزارش شدند. نتایج ارزیابی انحلال تری­کلسیم فسفات[2] در محیط مایع PKV توسط جدایه­ها نشان داد که. سویه K1، K2، و K3 به ترتیب با میزان 706، 661 و 588 میلی­گرم در لیتر بیشترین حل­کنندگی فسفر را به خود اختصاص داده و با یکدیگر تفاوت معنی داری نداشتند. نتایج بررسی­ها نشان داد که در محیط جامد حاوی کربنات روی دو سویه K4 و K5 به ترتیب با مقادیر 910/0 و 850/0 نسبت هاله به کلونی تفاوت معنی­داری با دیگر سویه­ها داشته و بقیه سویه­ها شامل K1، K2، و K3 و K6 توانایی انحلال کربنات روی از خود نشان ندادند. همچنین انحلال اکسید روی در محیط جامد توسط سویه K4 با 11/2 نسبت هاله به کلونی بیشترین میزان انحلال اکسید روی و سویه­های K1، K2، و K3 و K6 قادر به انحلال اکسید روی در محیط جامد نبودند. سویه­های  K1، K2، K3 و K6 قادر به انحلال ترکیبات کم محلول اکسید و کربنات روی در محیط مایع نبودند. سویه K4 با 5/12 میلی­گرم در لیتر با بیشترین میزان انحلال کربنات روی در محیط مایع را داشت و همچنین بیشترین انحلال اکسید روی در محیط مایع به سویه K4 با 76/9 میلی­گرم در لیتر نسبت داده شد. با توجه به نتایج بدست آمده می‌توان از این جدایه‌ها در جهت افزایش کارایی گیاه‌پالایی این عنصر در خاک‌های آلوده استفاده کرد.



[1] .Minimum Inhibitory Concentration (MIC)


[2] .Tricalcium Phosphate (TCP)

کلیدواژه‌ها


عنوان مقاله [English]

Isolation, identification and plant growth promoting characteristics of Cu-tolerant bacteria in a contaminated soil

نویسندگان [English]

  • Z. Karimi 1
  • P. AbbaszadehDahaji 2
  • A. R. Akhgar 3
  • M Hamidpour 3
1 M.Sc. Student, Department of Soil Science, Vali-e-Asr University
2 Assistant professor, Department of Soil Science, Vali-e-Asr University
3 Associate professor, Department of Soil Science, Vali-e-Asr University
چکیده [English]

Plant inoculation with effective plant growth promoting and heavy metal resistant rhizobacteria is an efficient method to improve the phytoremediation efficiency and extraction of heavy metals from contaminated soils. With respect to this reason, astudy was conducted to isolate, identify and evaluate the growth stimulating traits of copper-resistant bacteria from a Cu-contaminated soil. Six rhizospheric soil samples of indigenous plants were collected in Sarcheshmeh Khatoonabad, Kerman province. After screening, six strains were selected. Phenotypic, biochemical and genetic identification data showed that five and one isolates were belong to Pseudomonas and Bacillus genera respectively. Then, the minimum inhibitory concentration of copper, siderophore production, auxin and hydrogen cyanide production, the ability to solubilize mineral phosphate in solid and liquid medium, the ability to solubilize Zn low soluble compounds in solid and liquid medium and production of ACC-deaminase were investigated. The results indicated that all of the isolates were resistant to various concentrations of copper. The maximum resistance to inhibitory concentration of copper was related to K4 and K5 strains at a concentration of 400 mg L-1 of copper. All of the strains were able to produce siderophore in CAS medium except for K6 strains and strain K5 was also the superior in siderophore production. Among the strains, K4 and K6 were not able to produce auxin. There was also a significant difference between auxin productions by isolates K1 with the other isolates, with the highest auxin production (2.07 mg L-1). The results also demonstrated that among the strains, only K4 strain was capable of producing hydrogen cyanide (degree: four=relatively high) and the other isolates were reported as non-production of hydrogen cyanide (degree: one). The results of Three Calcium Phosphate (TCP) solubilizing by isolates in PKV broth medium showed that K1, K2, and K3 strains had the most phosphorus solubility (706, 661 and 588 mg L-1) respectively and there wasn’t significant difference between them. The results demonstrated that K4 and K5 isolates with 0.910 and 0.850 halos to colony diameter had a significant difference with other isolates in solid medium containing ZnCO3 and the rest of the strains, K1, K2, K3 and K6 did not show the ability to solubilize the zinc carbonate as well. The solubilizing of solid zinc oxide in solid medium by K4 strain (halo:colony, 2.11) was the highest solubility of zinc oxide and K1, K2, K3 and K6 strains were not able to dissolve the zinc oxide in the solid medium. They did not show the ability to dissolve the low soluble zinc oxide nor the zinc carbonate compounds. K4 strain had the highest zinc carbonate solubility in the broth (12.5 mg L-1) and the highest solubility of zinc oxide in the broth was attributed to K4 strain (9.76 mg L-1). According to the results, these isolates can be used for enhancing phytoremediation efficiency of this element in contaminated soils.

کلیدواژه‌ها [English]

  • Auxin
  • Plant growth promoting rhizobacteria
  • Minimum Inhibitory Concentration
  • Siderophore
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