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Development and Implementation of Rapid and Specific Detection Techniques for Seed-Borne Pathogens of Leafy Vegetable Crops

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Detection and Diagnostics of Plant Pathogens

Part of the book series: Plant Pathology in the 21st Century ((ICPP,volume 5))

Abstract

The usage of commercial seeds produced in few facilities increases the potential for introducing and disseminating several pathogens into new production areas. Fast and sensitive diagnostic tools are necessary to screen seeds and transplants. Identifying the source of inoculum is of critical importance for effective disease management. Due to the role of seed transmission, detection methods are also important for the production of pathogen-free seeds and for their certification. Pathogen’s detection on seeds is difficult because in most cases infected seeds can be asymptomatic, making visual detection difficult or impossible. Moreover, infected seeds may be present in a limited percent, and not evenly distributed within a lot. The development of the Polymerase Chain Reaction opened new perspectives in plant pathogen’s detection and today the most used assays are based on this technique or on new techniques developed in order to improve its sensitivity. This chapter focuses on recent developments for the quick detections of pathogens of leafy vegetables of recent introduction. During the past 15 years, several new diseases were introduced, in Italy as well as in many other countries, in this production sector through infected seeds. The molecular methods developed for the detection of several formae speciales of Fusarium oxysporum, Verticillium dahliae and several causal agents of foliar diseases of leafy vegetables are described and critically discussed.

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References

  • Agarwal VK (2006) Seed health. International Book Distributing, Lucknow

    Google Scholar 

  • Appel DJ, Gordon TR (1995) Intraspecific variation within populations of Fusarium oxysporum based on RFLP analysis of the intergenic spacer region of the rDNA. Exp Mycol 19:120–128

    Article  PubMed  CAS  Google Scholar 

  • Atkins SD, Clark IM, Sosnowska D, Hirsch PR, Kerry BR (2003) Detection and quantification of Plectosphaerella cucumerina, a potential biological control agent of potato cyst nematodes, by using conventional PCR, real-time PCR, selective media, and baiting. Appl Environ Microbiol 69:4788–4793

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Brouwer M, Lievens B, van Hemelrijk W, van den Ackerveken G, Cammue BPA, Thomma BPHJ (2003) Quantification of disease progression of several microbial pathogens on Arabidopsis thaliana using real-time fluorescence PCR. FEMS 228:241–248

    Article  CAS  Google Scholar 

  • Chiocchetti A, Sciaudone I, Durando F, Garibaldi A, Migheli Q (2001) PCR detection of Fusarium oxysporum f. sp. basilici on basil. Plant Dis 85:607–611

    Article  CAS  Google Scholar 

  • Ciccarese F (1987) Severe outbreaks of verticillium wilt on Cichorium intybus and Brassica rapa and pathogenic variations among isolates of Verticillium dahliae. Plant Dis 71:1144–1145

    Article  Google Scholar 

  • Du Toit LJ, Derie ML, Hernandez-Perez P (2005) Verticillium wilt in spinach seed production. Plant Dis 89:4–11

    Article  Google Scholar 

  • Duressa D, Rauscher G, Koike ST, Mou B, Hayes RJ, Marutchachalam K, Subbarao KV, Klosterman SJ (2011) A real-time PCR assay for detection and quantification of Verticillium dahliae in spinach seed. Phytopathology 102:443–451

    Article  Google Scholar 

  • Fujinaga M, Ogiso H, Shinohara H, Tsushima S, Nishimura N, Togowa M, Saito H, Nosue M (2005) Phylogenetic relationships between the lettuce root rot pathogen Fusarium oxysporum f. sp. lactucae race 1, 2, and 3 based on the sequence of the intergenic spacer region of its ribosomal DNA. J Gen Plant Pathol 71:402–407

    Article  CAS  Google Scholar 

  • Gachon C, Saindrenan P (2004) Real-time PCR monitoring of fungal development in Arabidopsis thaliana infected by Alternaria brassicicola and Botrytis cinerea. Plant Physiol Biochem 42:367–371

    Article  PubMed  CAS  Google Scholar 

  • Garibaldi A (1966) Due nuove malattie della valerianella. In: Proceedings 1st Congress Mediterranean Phytopathological Union, Bari-Napoli, Italy, pp 153–156

    Google Scholar 

  • Garibaldi A, Gullino ML (2010) Emerging soilborne diseases of horticultural crops and new trends in their management. Acta Hortic 883:37–46

    Google Scholar 

  • Garibaldi A, Gullino ML, Minuto G (1997) Diseases of basil and their management. Plant Dis 81:124–132

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Gullino ML (2002) First report of Fusarium oxysporum on lettuce in Europe. Plant Dis 86:1052

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Gullino ML (2003) First report of Fusarium oxysporum on Eruca vesicaria and Diplotaxis sp. in Europe. Plant Dis 87:201

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Gullino ML (2004a) Seed transmission of Fusarium oxysporum f. sp. lactucae. Phytoparasitica 32:61–65

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Gullino M (2004b) First report of Fusarium oxysporum causing vascular wilt of Lamb’s lettuce (Valerianella olitoria) in Italy. Plant Dis 88:83

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Pasquali M, Keiji S, Gullino ML (2004c) Seed transmission of Fusarium oxysporum of Eruca vesicaria and Diplotaxis muralis. J Plant Dis Prot 111:345–350

    Google Scholar 

  • Garibaldi A, Gilardi G, Gullino ML (2007) First report of Verticillium wilt caused by Verticillium dahliae on lettuce in Italy. Plant Dis 91:770

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Troisi M, Gullino ML (2009) First report of Fusarium wilt of endive (Cichorium endivia) caused by Fusarium oxysporum. Plant Dis 10:1,078.3

    Google Scholar 

  • Garibaldi A, Gilardi G, Poli A, Gullino ML (2011a) First report of Fusarium wilt of chicory (Cichorium intybus) caused by Fusarium oxysporum in Italy. Plant Dis 95:496

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Bertoldo C, Gullino ML (2011b) First report of a leaf spot of sweet basil (Ocimum basilicum) caused by Alternaria alternata in Italy. J Plant Pathol 93(supp 4s):4.71

    Google Scholar 

  • Garibaldi A, Gilardi G, Bertoldo C, Gullino ML (2011c) First report of leaf spot of wild (Diplotaxis tenuifolia) and cultivated (Eruca vesicaria) rocket caused by Alternaria japonica in Italy. Plant Dis 95:1316

    Article  Google Scholar 

  • Garibaldi A, Gilardi G, Ortu G, Gullino ML (2012) First report of Plectosphaerella cucumerina on wild rocket (Diplotaxis tenuifolia) in greenhouses in Italy. Plant Dis 96:1825

    Article  Google Scholar 

  • Guillemette T, Iacomi-Vasilescu B, Simoneau P (2004) Conventional and real-time PCR-based assay for detecting pathogenic Alternaria brassicae in cruciferous seed. Plant Dis 88:490–496

    Article  CAS  Google Scholar 

  • Guo JR, Schnieder F, Beyer M, Verreet JA (2005) Rapid detection of Mycosphaerella graminicola in wheat using reverse transcription-PCR assay. J Phytopathol 153:674–679

    Article  CAS  Google Scholar 

  • Hampton R, Ball E, De Boer S (eds) (1990) Serological methods for detection and identification of viral and bacterial plant pathogens: a laboratory manual. APS Press, St. Paul

    Google Scholar 

  • Henson JM, French R (1993) The polymerase chain reaction and plant disease diagnosis. Annu Rev Phytopathol 31:81–109

    Article  PubMed  CAS  Google Scholar 

  • Inderbitzin P, Davis RM, Bostock RM, Subbarao KV (2011) The ascomycete Verticillium longisporum is a hybrid and a plant pathogen with an expanded host range. PLoS One 6:18260

    Article  Google Scholar 

  • Katan T, Gamliel A, Katan J (1996) Vegetative compatibility of Fusarium oxysporum from sweet basil in Israel. Plant Pathol 45:656–661

    Article  Google Scholar 

  • Konstantinova P, Bonants PJM, van Gent-Pelzer MPE, van der Zouwen P, van der Bulk R (2002) Development of specific primer for detection and identification of Alternaria spp. in carrot material by PCR and comparison with blotter and plating assay. Mycol Res 106:23–33

    Article  CAS  Google Scholar 

  • Lievens B, Claes L, Vakalounakis DJ, Vanachter ACRC (2007) A robust identification and detection assay to discriminate the cucumber pathogens Fusarium oxysporum f. sp. cucumerinum and f. sp. radicis-cucumerinum. Environ Microbiol 9:2145–2161

    Article  PubMed  CAS  Google Scholar 

  • Lievens B, Rep M, Thomma BPHJ (2008) Recent developments in the molecular discrimination of formae speciales of Fusarium oxysporum. Pest Manag Sci 64:781–788

    Article  PubMed  CAS  Google Scholar 

  • Lievens B, Hanssen IM, Rep M (2012) Recent developments in the detection and identification of formae speciales and races of Fusarium oxysporum: from pathogenicity testing to molecular diagnosis. In: Gullino ML, Katan J, Garibaldi A (eds) Fusarium wilts of greenhouse crops. APS Press, St. Paul, pp 47–55

    Google Scholar 

  • Matsuda Y, Sameshima T, Moriura N, Inoue K, Nonomura T, Kakutani K, Nishimura H, Kusakari S, Tamamatsu S, Toyoda H (2005) Identification of individual powdery mildew fungi infecting leaves and direct detection of gene expression of single conidium by polymerase chain reaction. Phytopathology 95:1137–1143

    Article  PubMed  CAS  Google Scholar 

  • Mbofung GC, Pryor BM (2010) A PCR-based assay for detection of Fusarium oxysporum f. sp. lactucae in lettuce seed. Plant Dis 94:860–866

    Article  CAS  Google Scholar 

  • Mbofung GY, Hong SG, Pryor BM (2007) Phylogeny of Fusarium oxysporum f. sp. lactucae inferred from mitochondrial small subunit, elongation factor 1-α, and nuclear ribosomal intergenic spacer sequence data. Phytopathology 97:87–98

    Article  PubMed  CAS  Google Scholar 

  • McLaughlin RJ, Chen TA (1990) ELISA methods for plant pathogenic prokaryotes. In: Hampton R, Ball E, DeBoer S (eds) Serological methods for detection and identification of viral and bacterial plant pathogens. APS Press, St. Paul, pp 197–204

    Google Scholar 

  • Mullis KB, Faloona FA (1987) Specific synthesis of DNA in vitro via a polymerase catalyzed chain-reaction. Method Enzymol 155:335–350

    Article  CAS  Google Scholar 

  • Murakishi HH (1951) Purple seed stain of soybean. Phytopathology 41:305–318

    Google Scholar 

  • O’Donnell K, Kistler HC, Cigelink E, Ploetz RC (1998) Multiple evolutionary origins of the fungus causing Panama disease of banana: concordant evidence from nuclear and mitochondrial gene genealogies. Proc Natl Acad Sci U S A 95:2044–2049

    Article  PubMed  PubMed Central  Google Scholar 

  • Pasquali M, Piatti P, Gullino ML, Garibaldi A (2006) Development of a real-time polymerase chain reaction for the detection of Fusarium oxysporum f. sp. basilici from basil seed and roots. J Phytopathol 154:632–636

    Article  CAS  Google Scholar 

  • Pasquali M, Saravanakumar D, Gullino ML, Garibaldi A (2008) Sequence specific amplified polymorphism (SSAP) technique to analyse VCG 0300 of Fusarium oxysporum f. sp. lactucae. J Plant Pathol 90:527–535

    CAS  Google Scholar 

  • Pavón MÁ, Gonzáles I, Martín R, García T (2012) A real-time reverse-transcriptase PCR technique for detection and quantification of viable Alternaria spp. in foodstuff. Food Control 28:286–294

    Article  Google Scholar 

  • Pellegrino C, Gilardi G, Gullino ML, Garibaldi A (2010) Detection of Phoma valerianellae in lamb’s lettuce seeds. Phytoparasitica 38:159–165

    Article  Google Scholar 

  • Poli A, Gilardi G, Spadaro D, Gullino ML, Garibaldi A (2012) Molecular characterization of Fusarium oxysporum f. sp. cichorii pathogenic on cichory (Cichorium intybus). Phytoparasitica 40:383–391

    Article  CAS  Google Scholar 

  • Pryor BM, Gilbertson RL (2001) A PCR assay for detection of Alternaria radicina on carrot seed. Plant Dis 85:18–23

    Article  CAS  Google Scholar 

  • Randall-Schadel BL, Bailey JE, Beute MK (2001) Seed transmission of Cylindrocladium parasiticum in peanut. Plant Dis 85:362–370

    Article  Google Scholar 

  • Sheridan GEC, Masters CI, Shallcross JA, Mackey BM (1998) Detection of mRNA by reverse transcription-PCR as an indicator of viability in Escherichia coli cells. Appl Environ Microb 4:1313–1318

    Google Scholar 

  • Skovgaard K, Nirenberg HI, O’Donnell K, Rosendahl S (2001) Evolution of Fusarium oxysporum f. sp. vasinfectum races inferred from multigene genealogies. Phytopathol 91:1231–1237

    Article  CAS  Google Scholar 

  • Smith OP, Peterson GL, Bec RJ, Schaad NW, Bonde M (1996) Development of a PCR-based method for identification of Tilletia indica, casual agent of Karnal bunt of wheat. Phytopathology 86:115–122

    Article  CAS  Google Scholar 

  • Summerell BA, Gunn LV, Bullock S, Tesoriero LT, Burgess LW (2006) Vascular wilt of basil in Australia. Australas Plant Pathol 35:65–67

    Article  CAS  Google Scholar 

  • Swart L, van Niekerk JM (2003) First record of Fusarium oxysporum f. sp. basilici occurring on sweet basil in South Africa. Australas Plant Pathol 32:125–126

    Article  Google Scholar 

  • Taylor E, Bates J, Kenyon D, Maccaferri M, Thomas J (2001) Modern molecular methods for characterization and diagnosis of seed-borne fungal pathogens. J Plant Pathol 83:75–81

    CAS  Google Scholar 

  • Walcott RR (2003) Detection of seedborne pathogens. HortTechnology 13:40–47

    Google Scholar 

  • Webb K, Gilardi G, Matic S, Gullino ML, Garibaldi A (2014) Development of a rapid in planta detection method for identifying Plectosphaerella cucumerina on wild rocket by real-time PCR. Eur J Plant Pathol (accepted, in press)

    Google Scholar 

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Acknowledgements

The results reported have received funding from the European Union’s Seventh Framework Programme (FP7/2007–2013) under grants agreement n° 261752 PlantFoodSec – Plant and Food Biosecurity, Network of Excellence and 311875 “TESTA – Seed health: development of seed treatment methods, evidence for seed transmission and assessment of seed health”.

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Authors and Affiliations

  1. AGROINNOVA, University of Torino, Grugliasco, Torino, Italy

    M. Lodovica Gullino

  2. Center of Competence Agroinnova, University of Torino, Via Leonardo da Vinci 44, 10095, Grugliasco, Italy

    Giovanna Gilardi, Giuseppe Ortu & Angelo Garibaldi

Authors
  1. M. Lodovica Gullino
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  2. Giovanna Gilardi
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  3. Giuseppe Ortu
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  4. Angelo Garibaldi
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Correspondence to M. Lodovica Gullino .

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Editors and Affiliations

  1. AGROINNOVA, University of Torino, Grugliasco, Torino, Italy

    Maria Lodovica Gullino

  2. Plant Research International, Wageningen UR, Wageningen, Utrecht, The Netherlands

    Peter J. M. Bonants

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Gullino, M.L., Gilardi, G., Ortu, G., Garibaldi, A. (2014). Development and Implementation of Rapid and Specific Detection Techniques for Seed-Borne Pathogens of Leafy Vegetable Crops. In: Gullino, M., Bonants, P. (eds) Detection and Diagnostics of Plant Pathogens. Plant Pathology in the 21st Century, vol 5. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-9020-8_11

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