تأثیر ریزوبیوم بر تولید گلومالین توسط Rhizophagus irregularis در همزیستی با گیاه شبدر تحت سطوح نیتروژن

نوع مقاله : مقالات پژوهشی

نویسندگان

1 دانشگاه تبریز

2 دانشگاه علوم پزشکی تبریز

چکیده

گلومالین یک ترکیب گلیکوپروتئینی ویژه است که توسط قارچ‌های راسته گلومرال از رده گلومرومایکوتا تولید می‌شود و نقش کلیدی در ذخیره کربن آلی و نیتروژن خاک دارد. همچنین در تشکیل خاکدانه‌های پایدار و استقرار جوامع غنی میکروبی در خاک نقش بسزایی دارد. آزمایشی در قالب طرح فاکتوریل در پایه بلوک‌های کامل تصادفی با سه تکرار طراحی شد و گیاه شبدر (Trifolium repense L.)با قارچ Rhizophagus irregularis و باکتری Rhizobium leguminosarum bv. Trifolii تلقیح شد. چهار سطح نیتروژن (0، 2, 6 و 10 میلی‌مولار به فرم نیترات) بوسیله محلول غذایی نیومن و رومهلد ایجاد شد. گلدان‌ها با محلول غذایی آبیاری شدند. گیاهان شبدر پس از 12 هفته برداشت شد. گلومالین در بستر شن (SG) و گلومالین ریشه‌ای (RG) پس از استخراج از خاک، به روش بردفورد اندازه‌گیری شد. با افزایش سطوح نیتروژن میزان SG به طور معنی‌داری کاهش یافت (01/0 p

کلیدواژه‌ها


عنوان مقاله [English]

The Effect of Rhizobium on Glomalin Production by Rhizophagus irregularis in Symbiosis with Clover Plant under Different Levels of Nitrogen

نویسندگان [English]

  • vahideh Shaabani Zenoozagh 1
  • Nasser Aliasgharzad 1
  • Jaffar Majidi 2
  • Roghaieh Hajiboland 1
  • Behzad Baradaran 2
  • Leili Aghebati-Maleki 2
1 Tabriz universsity
2 Tabriz University of Medical Sciences
چکیده [English]

Introduction: Glomalin is a specific glycoprotein produced by the fungi belonging to phylum Glomeromycota and plays a key role in soil carbon and nitrogen storage. This also has a significant role in the stable aggregates formation and establishment of microbial communities in soil. Assimilated plant C which is allocated to the mycorrhizal fungus, appears as a recalcitrant glycoprotein (glomalin) in cell walls of hyphae and spores. Considering global warming due to increasing greenhouse gases, this phenomenon cab be important in carbon sequestration and reducing CO2 in atmosphere. Chemical fertilizers can affect symbiotic relations of these fungi, which in turn affect glomalin production.
Materials and Methods: In a factorial completely randomized design with three replication, clover plants (Trifolium repense L.) were included with Rhizophagus irregularis and/or Rhizobium leguminosarum bv. Trifolii. Four levels of nitrogen (0, 2, 6 and 10 mM as nitrate) in Newman & Romheld nutrient solution were applied to the pots containing 1.5 kg sterile sand. The pots were daily irrigated with nutrient solution containing the above-mentioned levels of nitrogen. Clover plants were excised after 12 weeks of growth. Fine roots were cleaned with %10 KOH and then stained using lactoglycerol trypan blue. Root colonization percentage was determined by grid line intersections method (GLM) described by Norrif et al (1992). For glomalin extraction, hyphal or root samples were autoclaved at 121 ⁰C with 50 mM sodium citrate buffer for 60 min in three cycles. Sand glomalin (SG) and root glomalin (RG) were measured by Bradford method after extraction. Nitrogen concentration in shoot and root was measured according to the standard method.
Results and Discussion: By increasing nitrogen level, the SG significantly decreased (p < 0.01), and at 2 mM, a 63.5 % decrease in SG was observed with relative to the nitrogen-free control. In the rhizobial treated pots, SG production increased by fungal inoculation (p < 0.01). The interaction between bacteria and AM was also significant in production of SG. At the presence of rhizobium bacteria, glomalin production by AM fungi increased significantly. The changes of glomalin content were not impacted by the presence of bacteria in the uninoculated pots with fungi. The highest amount of SG was recorded in the co-inoculated plants with nitrogen-free level. The amount of RG enhanced by increasing nitrogen concentration in nutrient solution. At 10 mM, RG increased by 12.90 %, 11.91 % and 1.44 % compared to the levels of 0, 2 and 6 mM, respectively. As the nitrogen level increased, the percentage of root colonization increased with respect to the control. Nitrogen concentration in shoot and root was enhanced by N increment to 10mM.
Conclusion: Carbon sequestration via glomali synthase by AM fungi is an important pathway for capturing CO2 from atmosphere. Field management measures help AM development of glomalin production. Based on our results, co-inoculated plants with AM and rhizobuim seem to positively affect the production of this glycoprotein. On the other hand, SG decreased significantly by increasing nitrogen concentrations in the nutrient solution. RG, however, increased significantly as a result of increased nitrogen in both fungal inoculations. The highest amount of RG was recorded in the co-inoculated plants with 10mM level. Glomalin synthesis by the fungi is positively affected by the soil nitrogen availability. Nitrogen is the main constituent of this glycoprotein. Plant photosynthates are translocated to the fungal organs via roots and mainly utilized for glomalin synthesis in hyphal and spore cell walls. During this process, nitrogen plays an important role as a constituent of the glycoprotein. The Bradford method was used for glomalin determination in this study. The method is not specific for glomalin and can also measure other glomalin related proteins and glycoproteins. Other proteins increased by N fertilization can hence be measured based on Bradford method. Once plant assimilates are translocated to the fungi, they may be transformed to the nitrogenous compounds if sufficient nitrogen sources are available. Accordingly, a considerable amount of fixed carbon is assimilated in fungal organs and soil particles. It can be concluded that carbon sequestration by arbuscular mycorrhizal symbiosis in terrestrial ecosystems can be improved by N fertilization at optimum level. In addition, the presence of rhizobium bacteria can meet the nitrogen requirement of plants through biological stabilization of nitrogen.

کلیدواژه‌ها [English]

  • Arbuscular mycorrhizal fungi
  • Bradford
  • Glomalin
  • Nitrogen
  • Rhizobium
1- Aliasgharzad N., Afshari Z., and Najafi N. 2016. Carbon Sequestration by Glomeral Fungi in Soil Is Influenced by Phosphorus and Nitrogen Fertilization. International Journal on Advanced Science Engineering Information Technology. 6:2088-5334.
2- Aliasgharzadeh N., Saleh Rastin N., Towfighi H., and Alizadeh A. 2001. Occurrence of arbuscular mycorrhizal fungal in salin soils of Tabriz plain of Iran in relation to some physical and chemical properties of soil. Mycorrhiza 11:119-122.
3- Barea J.M., Pozo M.J., Azcòn R., and Azcon-Aguilar C. 2005. Microbial co-operation in the rhizosphere. Journal of Experimental Botany 56:1761-1778.
4- Bedini S, Pellegrino E., Avio L., Pellegrini S., Bazzoffi P., Argese E., and Giovannetti M. 2009. Changes in soil aggregation and glomalin-related soil protein content as affect by the arbuscular mycorrhizal fungi species Glomus mosseae and Glomus intraradices. Soil Biology and Biochemistry 41, 1491e1496.
5- Bhat M.I., Bangroo S.A., Tahir A., Yadav S.R.S., and Aziz M.A. 2011. Combined Effects of Rhizobium and Vesicular Arbuscular fungi on green gram (Vigna radiata L. Wilczek) under temperate conditions. Journal of Agriculture Science, 2: 17-20.
6- Bohrer G., Kagan-Zur V., Roth-Bejerano N., and Ward D. 2001. Effects of environmental variables on vesicular-arbuscular mycorrhizal abundance in wild populations of Vangueria infausta. Journal of Vegetation Science, 12: 279-288.
7- Cornejo P., Meier S., Borie G., Rillig M.C., and Borie F. 2008. Glomalin-related soil protein in a Mediterranean ecosystem affected by a copper smelter and its contribution to Cu and Zn sequestration. Science of the Total Environment, 406:154-160.
8- Driver J.D., Holben W.E., and Rillig M.C. 2005. Characterization of glomalin as a hyphal wall component of arbuscular mycorrhizal fungi. Soil Biology and Biochemistry 37:101–106.
9- Ferrera-Cerrato R., and Villerias S.J. 1985. The VA endomycorrhiza and its effect of the development of three arboreous legumes. In Proceeding of the Sixth North American Conference on Mycorrhizae held at ben, Oregon, USA, pp. 328.
10- Garbaye J. 1994. Helper bacteria: a new dimension to the mycorrhizal symbiosis. New Phytologist. 128:197-210.
11- Garcia O.M., Ovasapyan T., Greas M., and Treseder K.K. 2008. Mycorrhizal dynamics under elevated CO2 and nitrogen fertilization in a warm temperate forest. Plant Soil 303: 301-303.
12- Gonzalez-Chavez M.C., Carrillo-Gonzalez R., Wright S.F., and Nichols K.A. 2004. The role of glomalin, a protein produced by arbuscular mycorrhizal fungi, in sequestering potentially toxic elements. Environmental Pollution, 130: 317-323.
13- Gryndler M., Larsen J., Hrselova H., Rezacova V., Gryndlerova H. et al. 2006. Organic and mineral fertilization, respectively, increase and decrease the development of external mycelium of arbuscular mycorrhizal fungi in a long-term field experiment. Mycorrhiza 16:159–166.
14- Hajiboland R., Rahmat S., Aliasgharzad N., and Hartikainen H. 2015. Selenium-induced enhancement in carbohydrate metabolism in nodulated alfalfa (Medicago sativa L.) as related to the glutathione redox state. Soil Science and Plant Nutrition ,61:676-687.
15- Harper J.E., and Gibson A.H. 1984. Differential nodulation tolerance to nitrate among legume species. Crop Science. 24: 497-801.
16- Huo L., Wu T.Y., Lin H.M., Cao S.Y., and Tang W.X. 2008. Effects of long-term fertilization on water-stable aggregates in calcic kastanozem of Loess Plateau. Chinese Journal of Applied Ecology. 19:545–550.
17- Kabir Z., O'Halloran I.P., Fyles J.W., and Hamel C. 1997. Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization: Hyphal density and mycorrhizal root colonization. Plant Soil 192:285–293
18- Kormanic P.P., and M.c. Graw. 1982.Quantification of VA mycorrhizae in plant roots. In Schenck NC (eds). Methods and Principles of Mycorrhizal Research . American Phytopathological Society. Saint Paul Minnesota. Pp: 37-45.
19- Li M., Liu R., Christie P., and Li X. 2005. Influence of three arbuscular mycorrhizal fungi and phosphorus on growth and nutrient status of Taro. Communication on Soil Science and Plant analysis 36: 2383-2396.
20- Liu R., Li M., and Meng X. 2000. Effects of AM fungi on endogenous hormones in corn and
21- cotton plants. Mycosystem 19: 91-96.
22- Lovelock C.E., Wright S.F., Clark D.A., and Ruess R.W. 2004. Soil stocks of glomalin produced by arbuscular mycorrhizal fungi across a tropical rain forest landscape. Journal of Ecology, 92: 278-287.
23- Lucy M., Reed E., and Glick B.R. 2004. Application of free living plant growth promoting rhizobacteria. Antonie van Leeuwenhoek Journal of Microbiology 86:1-25.
24- Miller M.H. 2000. Arbuscular mycorrhiza and phosohorus nutrition of maize; a review of Guelph studies. Canadian Journal of Plant Science 80: 47-52.
25- Newman G., and Romheld. 1999. Root excertion of carboxylic acids and protons in phosphorus-deficient plants. Plant and soil, 211: 121-130.
26- Nichols K.A., and Wright S.F. 2004. Contributions of soil fungi to organic matter in agricultural soils. In Magdoff F, Weil R (eds) Functions and management of soil organic matter in agroecosystems. CRC Press, Boca Raton, FL, Pp: 179–198.
27- Nichols K.A., and Wright S.F. 2006. Carbon and nitrogen in operationally defined soil organic matter pool. Biology and Fertility of Soils. 43: 215-220.
28- Norrif I.R., Read D.J., and Varma A.K. 1992. Methods in Microbiology Techniques for Study of Mycorrhiza. Academic press, London.
29- Peix B., Mazurier S., Lemanceau P., Siblot S., Berta G., Mougel C., and Van Tuinen D. 2007. Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots. Ney Phytologist, 176: 197-210.
30- Purin S., and Rillig M.C. 2007. The arbuscular mycorrhizal fungal protein glomalin: Limitations, progress, and a new hypothesis for its function. Pedobiologia, 51: 123-130.
31- Rillig M.C., Ramsey P.W., Morris S., and Paul E.A. 2003. Glomalin, an arbuscular mycorrhizal fungal soil protein, responds to land-use change. Plant and Soil. 253: 293–299.
32- Rillig M.C., and Mummey D.L. 2006. Mycorrhizas and soil structure. New Phytologist, 171: 41-53.
33- Rillig M.C., Wright S.F., Nichols K.A., Schmidt W.F., and Torn M.S. 2001. Large contribution of arbuscular mycorrhizal fungi to soil carbon pools in tropical forest soils. Plant and Soil, 233: 167-177.
34- Schindler F.V., Mercer E.R., and Rice J.A. 2007. Chemical characteristics of glomalin related soil protein (GRSP) extracted from soils of varying organic matter content. Soil Biology and Biochemistry, 39: 320-329.
35- Singh P.K. 2012. Role of glomalin related soil protein produced by arbuscular mycorrhizal fungi: a review. Agriculture Science Research Journal 2: 119–125.
36- Steinberg P.D., and Rillig M.C. 2003. Differential decomposition of arbuscular mycorrhizal fungal hyphae and glomalin. Soil Biology and Biochemistry, 35: 191-194.
37- Streeter J. 1988. Inhibition of legume nodule formation and N2 fixation by nitrate. CRC Critical Reviews in Plant Sciences, 7 : 1-24.
38- Swift M., and Bignell D. 2001. Standard methods for assessment of soil biodioversity and land use practice. International centre for research in Agroforestry (ICRAF) Southeast Asia. Available at: http://www.icraf.cgiar.org/sea. 2001 (visited 31 January 2018).
39- Talaat N.B., and Abdallah M.A. 2008. Response of faba bean (Vicia faba L.) to dual inoculation with Rhizobium and VA mycorrhiza under different levels of N and P fertilization. Journal of Applied Science Research, 4:1092 -1102.
40- Treseder K.K., and Allen M.F. 2000. Mycorrhizal fungi have a potential role in soil carbon storage under elevated CO2 and nitrogen deposition. New Phytologist 147:189–200.
41- Treseder K.K., Turner K.M., and Mack M.C. 2007. Mycorrhizal responses to nitrogen fertilization in boreal ecosystems: potential consequences for soil carbon storage. Global Change Biology 13: 78–88.
42- Violi H.A., Treseder K.K., Menge J.A., Wright S.F., and Lovatt, C.J. 2007. Density dependence and interspecific interactions between arbuscular mycorrhizal fungi mediated plant growth, glomalin production, and sporulation. Canadian Journal of Botany, 85: 63-75.
43- Walley F.L., and Germida J.J. 1997. Response of spring wheat (Triticum aestivum) to interactions between Pseudomonas species and Glomus clarum NT4. Biology and Fertility of Soils, 24: 365-371.
44- Wery J., Ture O., and Salsae L. 1986. Relationship between growth, nitrogen fixation and assimilation in a legume (Medicago sativa L.). Plant and Soil, 96:17-29.
45- Wilson P.W. 1940. The biochemistry of symbiotic nitrogen fixation. The University of Wisconsin Press, Madison, Wis.
46- Wilson G.W.T., Rice C.W., Rillig M.C., Springer A., and Hartnett D.C. 2009. Soil aggregation and carbon sequestration are tightly correlated with the abundance of arbuscular mycorrhizal fungi: Results from long-term field experiments. Ecology Letters, 12:452–461.
47- Wright S.F. 2000. A fluorescent antibody assay for hyphae and glomalin from arbuscular mycorrhizal fungi. Plant and Soil 226:171–177.
48- Wright S.F., and Upadhyaya A. 1996. Extraction of an abundant and unusual protein from soil and comparison with hyphal protein of arbuscular mycorrhizal fungi. Soil Science 161: 575-586.
49- Wright S.F., and Upadhyaya A. 1998. A survey of soils for aggregate stability and glomalin, a glycoprotein produced by hyphae of arbuscular mycorrhizal fungi. Plant and Soil, 198: 97-107.
50- Wright S.F., Franke-Snyder M., Morton J.B., and Upadhyaya A. 1996. Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots. Plant and Soil, 181: 193-203.
51- Wright S.F., Franke-Snyder M., Morton J.B., and Upadhyaya A. 1996. Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots. Plant and Soil 181: 193-203.
52- Xie Z-P., Staehelin C., Vierheilig H., Wiemken A., Jabbouri S., Broughton W.J., Vögeli-Lange R., and Boller T. 1995. Rhizobial nodulation factors stimulate mycorrhizal colonization of nodulating and nonnodulating soybeans. Plant Physiology, 108:1519–1525.
53- Zhu Y., and Miller R.M. 2003. Carbon cycling by arbuscular mycorrhizal fungi in soil-plant systems. Trends in Plant Science, 8:407-409.