بررسی تأثیر گلومالین در تثبیت ریشه‏ ای سرب در گیاه شبدر همزیست با قارچ رایزوفاگوس ایرگولاریز

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 دانش آموخته دکتری دانشگاه تبریز و استادیار گروه علوم خاک، دانشگاه علوم کشاورزی و منابع طبیعی گرگان،

2 استاد گروه علوم خاک، دانشگاه تبریز

چکیده

سابقه و هدف: تثبیت فلزهای سمی در ریشه‏ گیاهان مایکوریزی از جمله ساز وکارهای حفاظتی قارچ مایکوریزا همزیست در پاسخ به تنش فلزی است. نقش گلومالین به عنوان گلیکوپروتئین اختصاصی دیواره هیف و اسپور قارچ‏های آربوسکولار مایکوریزا (Arbuscular Mycorrhizal Fungi) در ناپویایی فلزهای سمی و کاهش پیامدهای تنش بر گیاه میزبان می‏تواند قابل ملاحظه باشد. با این پیش‏فرض هدف این مطالعه بررسی نقش گلومالین تولیدی توسط قارچ رایزوفاگوس ایرگولاریز همزیست با گیاه شبدر در تثبیت ریشه‏ای سرب بود.
مواد و روش‏ها: آزمایشی گلدانی در آرایش فاکتوریل در قالب طرح بلوک کامل تصادفی شاکل دو فاکتور قارچ (با و بدون قارچ Rhizophagus irregularis) و سطوح سرب (0، 150، 300 و 450 میکرومولار Pb+2) در پنج تکرار طراحی گردید. جهت استخراج گلومالین، نمونه‏های ریشه با بافر سیترات سدیم 50 میلی‏مولار (8=pH) به مدت یک ساعت در دمای C˚ 121 و طی سه چرخه پی در پی اتوکلاو شدند. غلظت گلومالین در عصاره استخراجی با استفاده از روش الایزا توسط آنتی‏بادی مونوکلونال 11B32 تعیین گردید. بعد از رسوب گلومالین، مقدار سرب کمپلکس شده پس از هضم با اسید نیتریک غلیظ اندازه‏گیری شد. وزن خشک اندام‏هوایی و ریشه، درصد کلنیزاسیون ریشه، مقدار فسفر و سرب اندام‏هوایی و ریشه و کارآیی جذب، استخراج و انتقال سرب مورد ارزیابی قرار گرفت.
یافته‏ها: با افزایش سطح سرب، وزن خشک اندام‏هوایی و ریشه گیاهان مایکوریزی و غیرمایکوریزی کاهش یافت. وزن خشک اندام‏هوایی و ریشه در سطوح 150، 300 و 450 میکرومولار سرب، به ترتیب کاهش 2/11، 9/12، 3/18 و 5/7، 1/18 و 7/36 درصدی نسبت به سطح صفر سرب داشتند. وزن خشک اندام‏هوایی و ریشه در گیاهان مایکوریزی به ترتیب 9/24 و 2/34 درصد بیشتر از گیاهان غیرمایکوریزی بود. اثر قارچ بر جذب فسفر اندام‏هوایی و ریشه معنی‏دار گردید، جذب فسفر اندام‏هوایی و ریشه در گیاهان مایکوریزی به ترتیب 2/32 و 8/45 درصد بیشتر از گیاهان غیرمایکوریزی بود. سرب جذب شده در اندام‏هوایی و ریشه در سطوح مختلف سرب و در گیاهان مایکوریزی به‏ طور معنی‏داری بیشتر از گیاهان غیرمایکوریزی بود. بیشترین مقدار جذب سرب در اندام‏هوایی و ریشه، در تیمار مایکوریزی سطح 450 میکرومولار سرب بود که افزایش 5/46 و 7/80 درصدی نسبت به تیمار غیرمایکوریزی در این سطح داشتند. در سطوح 150، 300 و 450 میکرومولار سرب، کارآیی جذب گیاهی سرب در گیاهان مایکوریزی افزایش 00/8، 5/14 و 2/49 درصدی نسبت به گیاهان غیرمایکوریزی داشت. اثر سطوح مختلف سرب بر کارایی استخراج و انتقال گیاهی سرب معنی‏دار شد. با افزایش سطح سرب کارآیی استخراج گیاهی افزایش یافت، به‏طوری که سطح 450 میکرومولار نسبت به سطح 150 و 300 میکرومولار افزایش 3/69 و 8/27 درصدی داشت. با افزایش غلظت سرب، از کارایی انتقال گیاهی سرب از ریشه به اندام‏هوایی کاسته شد. درصد کلنیزاسیون ریشه با افزایش غلظت سرب تا سطح 300 میکرومولار سرب افزایش و سپس در سطح 450 میکرومولار کمی کاهش یافت، اما از نظر آماری تفاوت معنی‏دار بین سطوح 150، 300 و 450 میکرومولار سرب وجود نداشت. با افزایش غلظت سرب، تولید گلومالین ریشه و مقدار سرب تثبیت شده توسط آن نیز به ‏طور معنی‏داری افزایش یافت.
نتیجه‏گیری: کلنیزاسیون ریشه گیاه شبدر با قارچ رایزوفاگوس ایرگولاریز منجر به بهبود رشد و تغذیه فسفری گیاهان مایکوریزی نسبت به گیاهان غیرمایکوریزی در شرایط تنش سرب شد. جذب سرب در ریشه گیاه شبدر بیش‏تر از اندام‏هوایی بود، بنابراین، گیاه شبدر در تثبیت ریشه‏ای سرب نقش مؤثرتری داشت. همچنین مقدار سرب ریشه در گیاهان مایکوریزی بیشتر از گیاهان غیرمایکوریزی بود که می‏تواند به دلیل تثبیت سرب در ساختارهای قارچی ریشه باشد. افزایش تولید گلومالین در ریشه‏های مایکوریزی به عنوان ترکیب اصلی و مؤثر دیواره اسپور و هیف قارچ نقش مهم و کلیدی در تثبیت ریشه‏ای سرب در گیاهان مایکوریزی ایفا می‏کند.

کلیدواژه‌ها

عنوان مقاله [English]

The effect of glomalin on root stabilization of lead in clover plant inoculated with Rhizophagus irregularis fungus

نویسندگان [English]

  • Elham Malekzadeh 1
  • Nasser Aliasgharzad 2
1
2 Department of Soil Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
چکیده [English]

Background and objectives: Root stabilization of toxic metals by mycorrhizal plants is one of the protective mechanisms of symbiotic arbuscular mycorrhizal (AM) fungi in response to metal stress. Role of the glomalin as a specific glycoprotein of spore and hyphal cell wall of AM fungi can be remarkable in sequestration of toxic metals and reduction of stress effects on host plant. Considering this hypothesize, the study was conducted to investigate the role of glomalin produced by Rhizophagus irregularis fungus symbiosis of clover plant in root stabilization of Pb.
Materials and Methods: A pot culture experiment was performed as completely randomized block design by two factors including AM fungus (inoculated with R. irregularis and non-inoculated) and four levels of Pb+2 (0, 150, 300 and 450 µM) with five replications. For glomalin extraction, root samples were autoclaved at 121˚C with 50 mM sodium citrate buffer for 1 hr in three cycles. Glomalin concentration in the extracted samples were determined by ELISA method using monoclonal antibody 32B11. After precipitation of the glomalin and its digestion in concentrated nitric acid, Pb-sequestrated by the glomalin were measured. Shoot and root dry weights, root colonization percentage, shoot and root P and Pb contents, and plant uptake, extraction and translocation efficiency of Pb were assessed.
Results: Shoot and root dry weights of mycorrhizal (M) and non-mycorrhizal (NM) plants were decreased by increasing of Pb levels. At the levels of 150, 300 and 450 μm Pb, shoot and root dry weights were decreased by 11.2%, 12.9%, 18.3% and 7.5%, 18.1%, 36.7% compared to the control (0 μm Pb), respectively. Shoot and root dry weights of M plants were increased by 24.9% and 43.2% compared to the NM ones. P contents of shoot and root were affected by AM fungus, so that the shoot and root P contents of M plants were increased by 32.2% and 45.8 % compared to the NM ones. At different levels of Pb, shoot and root Pb contents in M plants significantly were higher than NM ones. Maximum contents of Pb of shoot and root were recorded at level of 450 µM Pb in M plants which were increased by 46.5% and 80.7% compared to the NM ones at the same level. At the levels of 150, 300 and 450 µM Pb, the uptake efficiency of Pb in M plants was increased by 8%, 14.5% and 80.7% compared to the NM ones at the same levels. Based on ANOVA results, Pb-extraction and translocation efficiency were affected by Pb treatments. Pb-extraction efficiency of plants was increased as Pb concentration increased, so that the content of Pb-extraction efficiency at 450 µM of Pb was increased by 69.3% and 27.8% compared to the 150 and 300 µM of Pb, respectively. Plant Pb-translocation efficiency from root to shoot was decreased as Pb concentration increased. The percentage of root colonization was increased as the Pb concentration increased up to 300 µM Pb and then was slightly decreased as the level of Pb rose from 300 to 450 µM, but there was no significant difference between the levels of 150, 300 and 450 µM Pb. Glomalin production in root and Pb sequestrated by glomalin was significantly increased as Pb concentration increased.
Conclusion: Root colonization of clover plant by R. irreqularis led to improved growth and phosphorus nutrition of M plants compared to the NM ones, under Pb stress condition. Pb uptake was greater in roots than in shoots, therefore the clover plants played a more effective role in root stabilisation of Pb.

کلیدواژه‌ها [English]

  • ELISA
  • Biostabilisation
  • Bioremediation
  • Toxic metal
  • AM fungus

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مجله مدیریت خاک و تولید پایدار
دوره 9، شماره 3
آذر 1398
صفحه 69-90

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