Biological Control of African Violets Root-Knot Disease by the Used of Extracellular Protease Bacillus

محورهای موضوعی : مجله گیاهان زینتی

Hadi Rahanandeh ¹ (Member of Department of Plant Pathology, Faculty of Agriculture, Rasht Branch, Islamic Azad University, Rasht, Iran)
Mahsa Moshayedi ² (Young Researchers and Elite Club, Rasht Branch, Islamic Azad University)

تاریخ دریافت : 1395/11/09 تاریخ پذیرش : 1396/04/13 تاریخ انتشار : 1396/09/10

کلید واژه: Biological control, African violet media, Root knot disease, &lt, i&gt, Bacillus&lt, /i&gt, ,

چکیده مقاله :

The present study explored the efficacy of Bacillus spp. and protease production for biocontrol of the root-knot nematode Meloidogyne javanica in African violet media. Among 100 bacterial isolates from various soils, the highest nematode mortality was observed for treatments with isolate GM-18, which was identified as Bacillus subtilis based on cultural and morphological characteristics and 16S rDNA sequencing analyses. The Bacillus spp. showed nematicidal activity 74.01 to 28.33. Strain GM18 showed the most inhibitor zone in medium containing casein (13 mm) and nematicidal activity in our tests targeting Meloidogyne javanica. The effect of different variables such as time (1-24 h), pH (7- 8.5), and temperature (25- 37 °C) on the protease production was checked. The maximum enzyme production was noted when B. subtilis GM18 was grown on mineral medium and incubated at 31°C for 12 h with initial pH = 8. Three protease inhibitors that had the greatest influence on EDTA were studied. Protease produced by B. subtilis GM18belonged to different groups.

چکیده انگلیسی:

در این تحقیق تاثیر Bacillus spp. و پروتئاز تولیدی آن­ها روی نماتد غده  (Meloidogyne javanica) در میزبان بنفشه آفریقایی مورد بررسی قرار گرفت. حدود 100 جدایه باکتریایی از خاک ها جداسازی شد. تاثیر همه‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌‌ی باکتری‌ها بر میزان مرگ و میر نماتدها مورد بررسی قرار گرفت. از باکتری­های موثر چهار جدایه متعلق به جنس باسیلوس بودند. از باسیلوس های جداسازی شده جدایه GM18 بیشترین تاثیر را در مرگ و میر نماتد غده داشت. بر اساس آزمایش تولید پروتئاز 4 جدایه قادر به تولید پروتئاز در سطح پتری دیش بودند. بزرگترین هاله را جدایه GM18 به میزان 13 میلی متر تولید کرد. این جدایه بر اساس آزمایش­های بیوشیمیایی و توالی­سنجی B. subtilis تشخیص داده شد. فاکتورهای تاثیرگذار در میزان تولید پروتئاز همانند pH، دما، زمان و همچنین بازدارنده های تولید پروتئاز در مورد این جدایه بررسی گردید. نتایج آزمایش­­ها نشان داد که بیشترین میزان تولید پروتئاز در دمای 31 درجه سانتی گراد،  8=pH  و زمان 12 ساعت رخ می­دهد. بیشترین تاثیر بازدارندگی را EDTA روی پروتئاز تولیدی داشت که نشان­دهنده‌ی وجود میزان زیاد متالوپروتئاز می­باشد، البته بازدارنده‌های دیگر نیز به میزان کمتر تاثیر داشتند که نشان می­دهد پروتئاز تولیدی این باکتری از چند گروه پروتئاز مختلف تشکیل شده است.

منابع و مأخذ:

Abbasi, N., Mohammadi Torkashvan, A. and Rahanandeh, H. 2014. Evaluation of mushroom compost

for the bio control root-knot nematode. International Journal of Biosciences, 5 (8): 147-153.

Ahman, J. 2000. Extracellular serine proteases as virulence factors in nematophagous fungi: Molecular 

characterization and functional analysis of the PII protease in Arthobotrys oligospora. Doctoral 

Dissertation, Lund University.

Ahman, J., Johansson, T., Olsson, M., Punt, P.J., van den Hondel, C.A.M.J.J. and Tunlid, A.S. 2002.

Improving the pathogenicity of a nematode trapping fungus by genetic engineering of a subtilisin

with nematotoxic activity. Applied and Environmental Microbiology., 689: 3408–3415.

Al-Shehri Abdulrahman, M.M. and Yasser, S. 2004. Production of protease produced by Bacillus

licheniformis isolated from Tihamet Aseer, Saudi Arabia. Pakistan Journal of Biological Sciences.,

7 (9): 1631-135.

Li Bin, L., Guan-lin, S.A. and Coosemans, J. 2005. Suppression of Meloidogyne javanica by antagonistic

and plant growth-promoting rhizobacteria. Journal of Zhejiang University Science, 6 (6): 496-501.

Camila, R., Da, S., Anderia, B.D. and Meire, L.L.M. 2007. Effect of the culture conditions on the

production of an extracellular protease by thermophilic Bacillus sp. and some properties of enzymatic

activity. Brazilian Journal of Microbiology, 38: 253-258. 

Chi, Z., Ma, C., Wang, P. and Li, H.F. 2007. Optimization of medium and cultivation conditions

for alkaline protease production by the marine yeast Aureobasidium pullulans. Bioresource

Technology, 98: 534-538.

Cox, G.N., Kusch, M. and Edgar, R.S. 1981. Cuticle of Caenorhabditis elegans: Its isolation and partial

characterization. The Journal of Cell Biology, 90: 7–17.

Fang, Z.D. 1998. Methods of plant pathology. Beijing: China Agriculture Press.

Folasade, M., Olajuyigbe, J.O. and Ajele, E. 2005. Production dynamics of extracellular protease

from Bacillus species. African Journal of Biotechnology, 4 (8): 776-779.

Gardener, B.B.M. 2004. Ecology of Bacillus and Paenibacillus spp. in agricultural systems. Phytopathology,

94: 1252–1258.

Haddar, A., Bougatef, A., Agrebi, R., Sellami-Kamoun, A. and Nasri, M. 2009. A novel surfactant-stable

alkaline serine-protease from a newly isolated Bacillus mojavensis A21 purification and

characterization. Process Biochemistry, 44: 29–35.

Huang, X.W., Tian, B.Y., Niu, Q.H., Yang, J.K., Zhang, L.M. and Zhang, K.Q. 2005. An extracellular

protease from Brevibacillus laterosporus G4 without parasporal crystal can serve as a pathogenic

factor ininfection of nematodes. Research in Microbiology, 156: 719–727.

Huang, X.W., Zhao, N.H. and Zhang, K.Q. 2004. Extracellular enzymes serving as virulence factors

in nematophagous fungi involved in infection of the host. Research in Microbiology, 155: 811–816.

Ikram-ul-Haq Hamid, M. 2006. Fuzzy logic control of bioreactor for enhanced biosynthesis of alkaline 

protease by an alkalophilic strain of Bacillus subtilis. Current Microbiology, 52: 149-152.

Jansson, H.B. and Nordbring-Hertz, B. 1988. Infection mechanisms in the fungus-nematode system.

In: Poinar GO Jr, Jansson HB (eds) Diseases of nematodes, vol. 2. CRC, Boca Raton, FL, pp

59–72.

Jiyeong, P., Yunhee, S. and Young, H.K. 2014. Biological control of Meloidogyne hapla using an

antagonistic bacterium. Plant Pathology Journal, 30 (3): 288–298.

Leger, R.J., Nelson, J.O. and Screen, S.E. 1999. The entomopathogenic fungus Metarhizium anisopliae

alters ambient pH, allowing extracellular protease production. Microbiology, 145: 2691–2699.

Lian, L.H., Tian, B.Y., Xiong, R., Zhu, M.Z., Xu, J. and Zhang, K.Q. 2007. Proteases from Bacillus:

A new insight into the mechanism of action for rhizobacterial suppression of nematode populations.

Letters in Applied Microbiology, 45 (3):262-269. 

Moreira, K.A., Porto, T.S., Teixeira, M.F.S., Porto, A.L.F. and Lima, Filho, J.L. 2003. New alkaline

protease from Nocardiopsis sp.: Partial purification and characterization. Process Biochemistry.,

39: 67-72.

Morton, C.O., Hirsch, P.R. and Kerry, B.R. 2004. Infection of plant-parasitic nematodes by nematophagous

fungi: A review of the application of molecular biology to understand infection processes and

to improve biological control. Nematology. 6: 161–170.

Niu, Q.H., Huang, XW., Zhang, L., Li, YX., Li, J., Yang, J.K. and Zhang, KQ. 2006. A neutral 

protease from Bacillus nematocida, another potential virulence factor in the infection against

nematodes. Archives of Microbiology, 185: 439–448.

Oka, Y., Koltai, H., Bar-Eyal, M., Mor, M., Sharon, E., Chet, I. and Spiegel, Y. 2000. New strategies

for the control of plant-parasitic nematodes. Pest Management Science, 56: 983–988.

Olajuyigbe, F.M. and Ajele, J.O. 2005. Production dynamics of extracellular protease from Bacillus

species. African Journal of Biotechnology, 4 (8): 776-779.

Prakasham, R.S., Subba, R.C. and Sarma, P.N. 2006. Green gram husk an inexpensive substrate

for alkaline protease production by Bacillus sp. in solid-state fermentation. Bioresource Technology,

97: 1449-1454.

Qureshi, A.S., Bhutto, M.A., Khushk, I. and Dahot, M.U. 2011.  Optimization of cultural conditions

for protease production by Bacillus subtilis EFRL 01. African Journal of Biotechnology, 10

(26): 5173-5181.

Rahanandeh, H., Khodakaramian, G., Hassanzadeh, N., Seraji, A. and Asghari, S.M. 2013. Evaluation

of antagonistic Pseudomonas against root lesion nematode of tea. International Journal of Biosciences,

3(3): 32-40. 

Rahanandeh, H. and Moshaiedy, M. 2014. Potency evaluation of Pseudomonas aeroginosae and

Pseudomonas fluorescens as biocontrol agents for root-knot nematodes in Iran. International

Journal of Biosciences, 4 (12): 222-228.

Schisler, D.A., Slininger, P.J., Behle, R.W. and Jackson, M.A. 2004. Formulation of Bacillus spp.

for biological control of plant diseases. Phytopathology, 94: 1267–1271.

Siddiqui, I.A., Haas, D. and Heeb, S. 2005. Extracellular protease of Pseudomonas fluorescens

CHA0, a biocontrol factor with activity against the root-knot nematode Meloidogyne incognita.

Applied and Environmental Microbiology, 71: 5646–5649.

Siddiqui, Z.A. and Mahmood, I. 1999. Role of bacteria in the management of plant parasitic nematodes:

A review. Bioresource Technology., 69: 167–179.

Steinegger, D., Watkins, J.E., Vidaver, A.M. and Baxendale, F.P. 1974. G74-182 caring for African

violets. historical materials from university of Nebraska-Lincoln extension. Paper 971.

Tian, B.Y., Li, N., Lian, L.H., Liu, J.W., Yang, J.K. and Zhang, K.Q. 2006. Cloning, expression

and deletion of the cuticle-degrading protease BLG4 from nematophagous bacterium Brevibacillus

laterosporus G4. Archives of Microbiology, 186: 297–305.

Tian, B., Yang, J. and Ke-Qin Zhang, K.Q. 2007. Bacteria used in the biological control of plant-parasitic 

nematodes: Populations, mechanisms of action, and future prospects. FEMS Microbiology

Ecology, 61: 197–213.

Tunlid, A., Rosén, S., Ek, B. and Rask, L. 1994. Purification and characterization of an extracellular

serine protease from the nematode-trapping fungus Arthrobotrys oligospora. Microbiology,

140:1687–1695.

van Demark, P.J. and Batzing, B.L. 1987. The microbes, an introduction toheir nature and importance.

1 st edn. San Francisco: Benjamin/Cummings.

Watanabe, K. and Hayano, K. 1993. Distribution and identification of proteolytic Bacillus species

in paddy field soil under rice cultivation. Canadian Journal of Microbiology, 41: 674 – 680.

Wei, J.Z., Hale, K., Carta, L., Platzer, E., Wong, C., Fang, S.C. and Aroian, R.V. 2003. Bacillus

thuringiensis crystal proteins that target nematodes. Proceedings of the National Academy of

Sciences USA., 100: 2760–2765.

Wellingta, C.A., Do, N. and Meire, L.L.M. 2004. Production and properties of an extracellular

protease from thermophilic Bacillus sp. Brazilian Journal of Microbiology, 35: 91-96.