PCR and Nanotechnology Unraveling Detection Problems of the Seed-borne Pathogen Cephalosporium maydis, the Causal Agent of Late Wilt Disease in Maize

Author's: Ahmed Mohamed Awad, Ibrahim Hassan El-Abbasi, Tahsin Shoala, Sahara Ahmed Youssef, Dalia Mohamed Shaheen, Gomaa Ahmed Amer
Corresponding Author: Tahsin Shoala      Email: tahsinshoala2000@gmail.com
Article Type: Research Article     Published: May. 31, 2019 Pages: 30-39
DOI:        Views 294       Downloads0

Abstract:

Late wilt disease of maize caused by the fungus Cephalosporium maydis is one of the most important fungal diseases in Egypt. The pathogen spreads through the remaining plant debris in soil and through apparently healthy seeds. Detection of the latent infection of C. maydis pathogen is of great importance step in disease management which commonly achieved by applying the standard seed health tests. The commonly used seed health tests in laboratories around the world are still lake of specificity, sensitivity, speed, simplicity, cost-effectiveness, and reliability of seed health methods. In the current study, Infection percentage of seed-borne C. maydis recorded 1.0 and 1.5% in 2015 and 2016, respectively from whole maize seed (Giza 2) when examined by the standard blotter method. Using the same detection method examining broken maize seeds of cv. Giza 2 gave 3.5 and 5% in 2015 and 2016, respectively. Maximum infection percentage obtained during the current study period was 6.5% which was gotten when Potato Dextrose Yeast Agar (PDYA) applied with broken maize seeds of the same cultivar. While the highest infection percentage recorded during the entire study was 11% which obtained from broken seeds of hybrid Sc 166 of season 2015 and incubated on PDYA. Relatively to other major fungi recorded, the deep freezing method used with whole seeds (Giza 2) gave the least records of C. maydis in 2015 and 2016; 0.0 and 1.2, respectively. On the other hand, PDYA method gave the highest record during the two successive years of investigation (18.2) obtained from 2016 broken seeds. PCR has many beneficial characteristics that make it highly applicable for detecting C. maydis from seeds. In PCR diagnosis study, the development of DNA extraction is one of the most important steps. These approaches include using Ag NPs and Fe NPs in DNA extraction method to enhance the quantity and purity of the DNA template for successful PCR assay.

Keywords:

Maize, seed-borne, late wilt, Cephalosporium maydis, nano-particles, PCR.

Citation:

Awad, A.M., El-Abbasi, I.H., Shoala, T., Youssef, S.A., Shaheen, D.M., Amer, G.A., 2019. PCR and Nanotechnology Unraveling Detection Problems of the Seed-borne Pathogen Cephalosporium maydis, the Causal Agent of Late Wilt Disease in Maize. Int. J. Nanotechnol. Allied Sci., 3(2): 30-39.

REFERENCES

Al-Dhabaan, F.A., Yousef, H., Shoala, T., Shaheen, J., El Sawi, J.Y., Farag, T., 2017.  Enhancement of fungal DNA templates and PCR amplification yield by three types of nanoparticles. J. Plant Protec. Res., 58(1): 7.

Ali, K., Shuaib, M., Ilyas, M., Hussain, F., Arif, M., Ali, S., Jang, N., Hussain, F., 2017. Efficacy of Various Botanical and Chemical Insecticides against Flea Beetles on Maize (Zea maize L.). PSM Vet. Res., 2(1): 6-9.

Al-Jobory, H.J., Mahmoud, A.L.E., Al-Mahdi, A.Y., 2017. Natural Occurrence of Fusarium Mycotoxins (Fumonisins, Zearalenone and T-2 toxin) in Corn for Human Consumption in Yemen. PSM Microbiol., 2(2): 41-46.

Booth, C., 1971. The ginus Fusarium, Kew, Surrey. Common wealth Mycol. Inst., Kew, Surrey, England, pp 237.

Choi, Y.W., Hyde, K.D., Ho, W.H., 1999. Single spore isolation of fungi. Fungal Diver., 3:29-38.

Dellaporta, S.L., Wood, J., Hicks, J.B., 1983. A Plant DNA Minipreparation: Version II . Plant Mol. Biol. Rep.,1: 19-21.

Drori, R., Sharon, A., Goldberg, D., Rabinovitz, O., Levy, M., Degani, O., 2013. Molecular diagnosis for Harpophora maydis, the cause of maize late wilt in Israel. Phytopathol.  Mediterr., 52(1): 16-29.

El-Abbasi, I.H., 1990. Studies on some seed-borne diseases of certain food crops in Egypt. M. Sc. Thesis, Faculty of Agriculture, Ain Shams University: 83 pp.

Hildebrand, E.M., 1938. Techniques for the isolation of single microorganisms. Bot.                      Rev., 4: 628- 658.

Iqbal, M.N, Ashraf, A., Wang, S., 2018. Gold Nanoparticles (GNPs): Synthesis, Properties and Conjugation Methods. Int. J. Nanotechnol. Allied Sci., 2(1): 1-2.

Ismaeel, S.H., Mabrouk, M.S., Ali, A.A., Elwalead, K.A., 2017. Synthesis and Characterization of Bentonite Nanocomposites from Egyptian Bentonitic Clay. Int. J. Nanotech. Allied. Sci., 1(1): 16-29.

ISTA., 2014. International Seed Testing Association, Annual Meeting 2015, Montevido, Uruguay.  ISTA. News. Bulletin., No. 148.

Khiyami, M.A., Almoammar, H., Awad, Y.M., Alghuthaymi, M.A., Abd- Elsalam K.A., 2014. Plant pathogen nano diagnostic techniques: forthcoming changes? Biotechnol. Biotechnol. Equipmen., 28(5): 775-785.

Mathur, S. B., Cunfer, B.M., 1993. Seed-borne diseases and seed health testing of wheat. Jordbrugsforlaget, Frederiksberg, Denmark., 168 pp.

McCarthy, J.R.; J. Bhaumik; M.R. Karver; S.S. Erdem and R. Weissleder (2010). Target nanoagents for the detection of cancers. Molec. Oncol., 4(6): 511–528.

Michail, S.H., Abou-Elseoud, M.S., Eldin, M.S.N., 1999. Seed health testing of corn for Cephalosporium maydis. Acta Phytopath. et Entomol. Hungarica., 34 (1/2): 35-41.

Nadeem, M.K., Qaswar, M., Ahmed, N., Rabnawaz, Rasool, S.J., 2017. Effect of Seed Soaking Time on Germination of Maize (Zea mays L.). PSM Biol. Res., 02(1): 46-50.

Neergaard, P., 1979. Seed Pathology. vol. 1and2. The Macmillan Press Ltd., London and Basingstoke., 1191 pp.

Pervaiz, Z.H., Turi, N.A.I., Rabbani, M.A., Malik S.A., 2011. A modified method for high-quality DNA extraction for molecular analysis in cereal plants. Ph.D. thesis Depart. Biochem., Quaid-e- Azam Univ. Islam., Pakistan.

Rai, M., Yadav, A., Gade, A., 2009. Silver nanoparticles as a new generation of antimicrobials. Biotechnol. Adv., 27(1): 76–83.

Rehman, A., Sarwar, Y., Raza, Z.A., Hussain, S.Z., Mustafa, T., Khan, W.S., Ghauri, M.A., Haque, A., Hussain, I., 2015. Metal nanoparticle assisted polymerase chain reaction for strain typing of Salmonella typhi. Analyst., 140 (21): 7366–7372.

Samra, A.S., Sabet, K.A., Hingoroni, M.K. 1962. A new wilt of maize in Egypt.  Pl. Dis. Rptr., 46: 481-483.

Samra, A.S., Sabet, K.A., Hingoroni, M.K., 1963. Late wilt disease of maize caused by Cephalosporium maydis. Phytopathol., 53: 402- 406.

Selim, H.M., Mohamed, D.S., Eskander, H.M.G., 2017. Silver Nanoparticles: Synthesis, Medical Application, and Toxicity Effects. Int. J. Nanotechnol. Allied. Sci., 1(1): 39-46.

Shurtleff, M.C., 1980. Compendium of corn diseases. The Amer. Phytopathol. Soc., St. Paul, Minnesota, Second ed. 105pp.

Tan, H., Huang, H., Tie, M., Ma, J., Li, H., 2013. Comparative analysis of six DNA extraction methods in Cowpea (Vigna unguiculata L. Walp). J. Agri. Sci., 5(7): 82-90.

Warham, E.J., 1990. Effect of Tilletia indica infection on viability, germination and vigour of wheat seed. Pl. Dis.,74: 130-132.

Zaynab, M., Noman, A., Fatima, M., Saleem, T., Abbas, S., Raza, A., Iqbal, M.N., 2018. Proteomics Approach Reveals Seed Germination Mechanism in Model Plants. PSM Microbiol., 3(1): 30-36.

Zhonghua, M., Michailides, T.J., 2007. Approaches for eliminating PCR inhibitors and designing PCR primers for the detection of phytopathogenic fungi. Crop Protection., 26: 145-161.