Evaluation of Antagonistic of the some Fungal isolates on Golden Potato Cyst Nematode (Globoderarostochiensis) in vitro and Greenhouse Conditions in Hamedan Province

Document Type : Research Article

Authors

• Kh. Abbasi ¹
• D. Zafari ²

¹ , Faculty of Agriculture, Bu-Ali Sina University,Hamedan, Iran

² Department of Plant Protection, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran

10.22067/jpp.v0i0.59117

Abstract

Introduction:Potato (Solanumtuberosum) is one of the most important crops used as a source of human food. Iran is the third-largest producer of potato in Asia, where the production rate in 2015 was estimated to be about 5 million tonnes. Potato producers inHamedan province produce 21.3% oftotal potato harvestedinIran. Golden potato cyst nematode, Globoderarostochiensis is the most destructive potato pathogen. As the chitin is a dominant composition in middle layer of the eggshell, using the chitinases produced as chitin-degrading enzymes in a wide range of fungiis a good strategy for biological control of the golden potato cyst nematode. We assessedthe ability of various antagonistic fungi to control Globoderarostochiensisunder in vitro and greenhouse conditions.
Materials and Methods: Thirty four fungal isolates obtained from infected eggs of the potato cyst nematode, Globoderarostochiensisin potato fieldsofHamedan were evaluated in two chitin-agar and water-agar mediums under in vitro and greenhouse conditions.The ability of thechitinase enzyme production was assessedin chitin-agar medium with colloidal chitin as substrate, so the chitin was used as exclusive source of carbon.Colloidal chitin was prepared based onthe procedure of Seyedasliet al. (2004) with 10 g of powder chitin from practical-grade crab shell chitin (Sigma) in 100 ml of 85% H3PO4. Water was added to the above mixtureand was filtered with cheese cloth. To completely remove acid, water addition and filtrationrepeated for several times. The produced unguent materialwas dried and powdered and then used as carbon source in the medium. 0.5 percent of colloidal chitin was added to the medium. Afterwards, a 5 mm disk from the edges of 5days old was placed in the center of Petridish and all of them were kept for 5 days at 25º C. Chitinase detection medium (chitin-agar) was directly supplemented with colloidal chitin (5 g/l) and bromocresol purple (0.15 g/l). The ability of antagonistic activity of the fungi on the cyst nematode was testedin water-agar medium through assessing the interaction between fungi and cysts. The numbers of healthy and parasitized (dead) larvae and eggs were calculated after two weeks. The ability of the antagonistic fungi under greenhouse conditions was also analyzed. To provide fungal inoculum, 20g of soaked wheat seed were cast in nylon with autoclave capability. 2 ml distilled water were added per gram of cast seed and they were autoclaved threetimesduring 24 hours. Four fungi disk with 5 mm diameter from selected isolates were then added to allnylonswiththreerepetitions and werekept in 25ºC and dark conditions. To colonizeall of the seeds and avoid hangingthem, the seeds in nylons were mixed within 48 hours interval. After three weeks all of the seeds were infected with fungal isolates. The ability of the antagonistic fungi under greenhouse conditions was studiedby adding fungal inoculum and 100 cysts to each pot and performance evaluation of potato traits in pot after 90 days.
ANOVA (Analysis of variance) data analysis was conducted using of the SAS software version 9.0 in completely randomized design (CRD) with three replicates under all conditions.
Results and Discussion:The ANOVA results ofqualitative evaluation for chitinase activity in Petri dish containing chitin-agar showed significant difference among isolates at1% level of significance. Furthermore, there was apositiveassociationbetween diameter and chitanase activity. Isolates 153(P. lilacinus) and 6 (C. parapsilosis) had the maximum and minimumdiameter, respectively.
A significant difference was foundamong isolatesin the 0.1% level of significance. Isolate 152 (L. muscarium) had the greatestantagonistic ability and 62 (F. solani)was the weakest antagonist isolate.
Mean comparisons of measured performance traits of potato in pot showed in all traits thatisolates 154 (T. atroviridae)and 151 (B. bassiana)were the best antagonist isolates under greenhouse conditions. Isolates56 (F. equiseti)and 12 (F. oxysporum)with the lowest measured values in all functional traits were alsothe weakest antagonist fungi.
Conclusions:The results illustrated a strong correlation between antagonistic ability of fungal isolates under in vitro and greenhouse conditions.Therefore, fungal isolates can effectively reduce the nematode damages. Finally, isolates 151(B. bassiana), 152(L. muscarium), 153 (P. lilacinus) and 154(T. atroviridae) were selected as the strongest antagonistic fungi in controlling this nematode.

Keywords

• Biological control
• Chitin-agar
• Parasitism
• Yield traits

20.1001.1.20084749.1396.31.2.14.3

---

References

1- Agrawal T., and Kotasthane A.S. 2012. Chitinolytic assay of indigenous Trichoderma isolates collected from different geographical locations of Chhattisgarh in Central India. Springer Plus, 1(73):1-10.

2- Ahmadi K., Gholizadeh H., Ebadzadeh H., Hosseinpour R. Hatami F., Fazli B., Kazemian A., and Rafiei M. 2015. Agricultural crops statistics. Ministry of Agriculture, Department of Planning and Economy, Center for Information and Communication Technology, 1:158.(in Persian(

3- Brown R. H., and Kerry B.R. 1987. Principles and practices of nematode control in crops. Academic press. New York.

4- Dackman C. 1990. Fungal Parasites of the Potato Cyst Nematode Globodera rostochiensis: Isolation and Reinfection. Journal of Nematology, 22(4):594-597.

5- Dong J. Y. Zhao Z. X. Cai L. Liu S. Q. Zhang H. R. Duan M. and Zhang K.Q. 2004. Nematicidal effect of fresh water fungal cultures against the pine-nematode, Bursaphelenchus xylophilus. Fungal Diversity, 15:125–135.

6- Gitty M. Tanha maafi Z. Arjmandian A. and Pishevar S. 2011. Occurrence of potato golden cyst nematode in Iran and its distribution in Hamadan province. Agricultural Biolotechnology, 10(1):53-61.(in Persian with English abstract (

7- Khezri Nezhad N. Niknam G. Ghoosta Y. and Parvizi R. 2004. Evaluation of the biodiversity of nematodes in sugar beet fields and natural infection of sugar beet cyst nematode with antagonistic fungi in West Azerbaijan province, Urmia university master's thesis. p 185.(in Persian with English abstract (

8- Liu t. Wang L. Duan Y. X. and Wang X. 2008. Nematicidal activity of culture filtrate of Beavaria bassiana against Meloidogyne hapla. World Journal of Microbiology and Biotechnology, 24:113-118.

9- Lopez-Lima D. Sanchez-Nava P. Carrion G. and Núñez-Sanchez A.E. 2013. 89 % reduction of a potato cyst nematode population using biological control and rotation. Agronomy for Sustainable Development, 33:425–431.

10- Mahdikhani Moghadam E. Rouhani H. and Falahi Rastgar M. 2009. Biological control of sugar beet cyst forming nematode with Trichoderma under in vitro and green house condition.Water and Soil Science (Journal of Science and Technology of Agriculture and Natural Resources), 13(48): 301–313.

11- Mahdikhani Moghadam E. and Rouhani H. 2012. Effect of isolates of Trichodermaharzianum, T. virens and Bacillus subtilis for controlling Heterodera schachtii in field conditions. Journal of Plant Protection,26(1):75–81.

12- Manzanilla-Lopez R. H., Esteves I. Fintti-Sialer M. M. Hirsch P. R. Ward E. Devonshire J. and Hidalgo-Diaz L. 2013. Pochonia chlamydosporia: Advances and challenges to improveIts performance as a biological control agent of sedentary endo-parasitic nematodes. Journal of Nematology, 45(1):1–7.

13- Morton C. O. Hisch P. R. and Kerry B.R. 2004. Infection of plant parasitic nematodes by nematoghagous fungi – a review of the application of molecular biology to understand infection processes and to improve biological control. Nematology, 6(2):161-170.

14- Safari Motlagh M. R. and Samimi Z. 2013. Evaluation of Trichoderma spp. as biological agents in some of plant pathogens. Annals of Biological Research, 4(3):173-179.

15- Saifullah. and Naqibullah khan. 2014. Low temperature scanning electron microscopic studies on the interaction of Globodera rostochiensis woll and Trichoderma harzianum rifai. Pakistan Journal of Botany, 46(1):357-361.

16- Saifullah. and Thomas B.J. 1996. Studies on the parasitism of Globodera rostochiensisby Trichoderma harzianum using low temperature scaning electron microscopy. International Journal of Nematology, 6:117-122.

17- Santos M. C., Esteves I. Kerry B. and Abrantes I. 2013. Biology, growth parameters and enzymatic activity of Pochonia chlamydosporia isolates from potato cyst and root-knot nematodes. Nematology, 15:493-504.

18- Seyedasli N. Zamani M. Motallebi M. and Harighi M.J. 2004. The study of chitinase production in Trichoderma fungus. Iran Biology, 17(3):227-246.(in Persian(

19- Sharon E. Bar-Eyal M. Chet I. Herrera E. Strella A. Klelfeld O. and Splege Y. 2001. Biological control of the root-knot nematode Meloidogyne javanicaby Trichoderma harzianum. Phytopathology, 91:687-693.

20- Sharon E., Chet I., and Spiegel Y. 2009. Improved attachment and parasitism of Trichoderma on Meloidogyne javanica in vitro. European Jornal of Plant Pathology, 123:291-299.

21- Tikhonov V. E., Lopez-Llorca L.V., Salinas J., and Jansson H.B. 2002. Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium. Fungal Genetics and Biology, 35:67-78.

22- Tobin J. D., Haydock P. J., Hare M. C., Woods S. R., and Crump D.H. 2008. Effect of the fungus Pochonia chlamydosporia and fosthiazate on the multiplication rate of potato cyst nematodes (Globodera pallid and G. rostochiensis) in potato crops grown under UK field conditions. Biological Control, 46:194–201.

23- Windham G. L., Windham M. T., and Williams W.P. 1989. Effects of Trichoderma spp. on maize growth and Meloidogynearenaria reproduction. Plant Disease 73:493-495.

24- Zarghani A., Fatemi S., and Mahdian S. 2014. Evaluation of the pathogenicity of golden nematode Globodera rostochiensis on potato. Plant Protection (Journal of Agriculture), 37(4):11-21.(in Persian(