Asmalarda fungal gövde hastalıklarının araştırılmasında farklı inokulasyon yöntemlerinin karşılaştırılması

Öz

Amaç: Asma gövde hastalıkları, birçok fungal bitki patojeninin neden olduğu kompleks hastalıklardır. Bu çalışmada dört farklı hastalığı (Petri ve Siyah Bacak Hastalığı, Botryosphaeria ve Phomopsis Geriye Ölüm Hastalıkları) temsil eden bazı fungal patojenler için en uygun inokulasyon yönteminin saptanması amaçlanmıştır.

Yöntem ve Bulgular: Phaeomoniella chlamydospora, Dactylonectria macrodidyma, Neofusicoccum parvum ve Diaporthe ampelina türü funguslar dormant asma kalemlerine veya yeşil sürgünlere farklı yöntemlerle inokule edilmişlerdir. Dormant kalemlere uygulanan yöntemler; a) kabuk altına miselyal agar disk inokulasyonu, b) tepeden agar disk inokulasyonu, c) ksileme enjeksiyon yöntemi ve d) yeşil sürgünlere agar disk inokulasyonudur. Patojenlerin kalemler ve yeşil sürgünlere inokulasyonunu takiben 2 ay süreyle gelişmelerine izin verilmiş ve sonra dokularda oluşan lezyon uzunlukları ölçülmüştür. Sonuçlara göre tüm patojenler tüm inokulasyon yöntemleriyle, hem dormant kalemler hem de yeşil sürgünler üzerinde lezyonlar meydana getirmiş ancak ölçülen lezyon uzunlukları kullanılan yöntemlere göre farklılık göstermiştir. En uzun lezyonların oluştuğu patojen ve inokulasyon metodu eşleşmesi şu şekilde gerçekleşmiştir; Diaporthe ampelina için yeşil sürgüne miseliyal agar disk inokulasyonu (72.2 mm), D. macrodidyma ve P. chlamydospora için ksileme enjeksiyon (sırasıyla 23.0 ve 66.1 mm) ve Neofusicoccum parvum için kabul altına miseliyal agar disk inokulasyonu (51.5 mm).

Genel Yorum: Asmalarda fungal gövde hastalıklarının araştırılmasında birçok araştırmacı tarafından farklı inokulasyon yöntemleri kullanılsa da, patojenisite testlerinin bu çalışmada denenen yöntemlerden herhangi biri ile yapılabileceği ortaya koyulmuştur. Çok sayıda spor üreten türlerle yapılan çalışmalarda, ksileme enjeksiyon yönteminin daha makul sonuçlar vereceği görülmüştür. Miselyal gelişimi güçlü ve piknit üreten türler için ise kabuk altına ya da tepeden agar disk inokulasyon yöntemlerinin daha uygun olduğu anlaşılmıştır.

Çalışmanın Önemi ve Etkisi: Bir araştırmada, inokulasyon yönteminin seçimi o araştırmanın sonuçlarını doğrudan etkilemektedir. Bu çalışmanın sonuçlarının, asma gövde hastalıkları ile yapılacak araştırmalarda uygun bir inokulasyon yönteminin seçimine yardımcı olabileceği düşünülmektedir.

Anahtar Kelimeler

• Asma
• Dactylonectria nacrodidyma
• Diaporthe ampelina
• Neofusicoccum parvum
• Phaeomoniella chlamydospora

Comparison of different inoculation methods in the investigation of fungal trunk diseases in grapevines

Abstract

Aims: Grapevine trunk disease are complex diseases caused by many fungal plant pathogens. In this study, it was aimed to determine the most suitable inoculation method for some fungal pathogens representing four different diseases (Petri and Black Foot, Botryosphaeria and Phomopsis Dieback).

Methods and Results: Fungal species Phaeomoniella chlamydospora, Dactylonectria macrodidyma, Neofusicoccum parvum and Diaporthe ampelina were inoculated into dormant canes or green shoots by different inoculation methods. These methods for dormant cuttings were as follows; mycelial agar disc inoculation under bark (a), mycelial agar disc placement onto the top of cuttings (b), xylem injection method (c) and mycelial agar disc inoculation on green shoots (d). After inoculation of the pathogens on cuttings and shoots, they were allowed to grow for 2 months and then lesion lengths on the tissues were measured. As a result, all the pathogens produced lesions on both dormant canes and green shoots with all inoculation methods, but measured lesion lengths differed from the inoculation methods used. The pathogen and inoculation methods matching, in which the longest lesions were formed, were as follows; mycelial agar disc inoculation on green shoots for Diaporthe ampelina (72.2 mm), xylem injection method for Dactylonectria macrodidyma and Phaeomoniella chlamydospora (23.0 mm and 66.1 mm respectively) and mycelial agar disc inoculation under bark for Neofusicoccum parvum (51.5 mm).

Conclusions: Although different inoculation methods have been used by many researchers to investigate in grapevine fungal trunk diseases, it has been revealed that pathogenicity tests can be performed using one of methods used in this study. In studies with a large number of spore producing species, it has been observed that the xylem injection method would yield more plausible results. It was also found that mycelial agar disc inoculations, under bark or on the top of the cuttings, were more suitable for the species having a strong mycelial growth and pycnidial production.

Significance and Impact of the Study: The selection of the inoculation method in a study directly affects the results of that research. It is thought that the results of this study may contribute to the selection of the appropriate inoculation method in researches on grapevine trunk diseases.

Keywords

• Grapevine
• Dactylonectria nacrodidyma
• Diaporthe ampelina
• Neofusicoccum parvum
• Phaeomoniella chlamydospora

Kaynakça

1. Abou-Mansour E, Debieux J, Ramirez-Suero M, Benard-Gellon M, Magnin-Robert M, Spagnolo A, Chong J, Farine S, Bertsch C, L’Haridon F, Serrano M, Fontaine F, Rego C, Larignon P (2015) Phytotoxic metabolites from Neofusicoccum parvum, a pathogen of Botryosphaeria dieback of grapevine. Phytochemistry 115: 207-215.
2. Alaniz S, Leon M, Vicent A, Garcia-Jimenez J, Abad-Compos M, Armengol J (2007) Characterisation of Cylindrocarpon species associated with black foot disease of grapevine in Spain. Plant Dis. 91: 1187-1193.
3. Amponsah NT, Jones EE, Ridgway HJ, Jaspers MV (2008) Production of Botryosphaeria species conidia using grapevine green shoots. N. Z. Plant Prot. 61:301-305.
4. Amponsah NT, Jones EE, Ridgway HJ, Jaspers MV (2012) Susceptibility of grapevine tissues to Neofusicoccum luteum conidial infection. Plant Pathol. 61: 719–729.
5. Anonymous (2013). Technical Report, COST 017/13. Brussels, Belgium: Memorandum of understanding for the implementation of a European concerted research action designated as COST Action FA1303: Sustainable control of grapevine trunk diseases.
6. Ayres M, Sosnowski M, Wicks T (2011) A rapid technique for evaluating treatments for Eutypa dieback kontrol. Wine and Viticulture J. 26: 50-53.
7. Baumgartner K, Fujiyoshi PT, Travodon R, Castlebury LA, Wilcox WF, Rolshausen PE (2013) Characterization of species of Diaporthe from wood cankers of grape in Eastern North American vineyards. Plant Dis. 97: 912-920.
8. Bertsch C, Ramirez-Suero M, Magnin-Robert M, Larignon P, Chong J, Abou-Mansour E, Spagnolo A, Clement C, Fontaine F (2013) Grapevine trunk diseases: complex and still poorly understood. Plant Pathol. 62: 243-265.

9. Booth C, Stover RH (1974) Cylindrocarpon musae sp. nov., commonly associated with burrowing nematode (Radopholus similis) lesions on bananas. Trans. Br. Mycol. Soc. 63: 503-507.
10. Burgess TI, Barber PA, Hardy GES (2005) Botryosphaeria spp. associated with eucalypts in Western Australia, including the description of Fusicoccum macroclavatum sp. nov. Australas. Plant Pathol. 34: 557–567.
11. Burruano S, Mondello V, Conigliaro G, Alfonzo A, Spagnolo A, Mugnai L (2008) Grapevine decline in Italy caused by Lasiodiplodia theobromae. Phytopathol. Mediterr. 47: 132-136.
12. Crous PW, Gams W (2000) Phaeomoniella chlamydospora gen. et comb. nov., a causal organism of Petri grapevine decline and esca. Phytopathol. Mediterr. 39: 112-118.
13. Gomez KA, Gomez AA (1984) Statistical Procedures for Agricultural Research. Wiley Interscience, Canada. 680p.
14. Gramaje D, Urbez-Torres JR, Sosnowski MR (2018) Managing grapevine trunk diseases with respect to etiology and epidemiology: current strategies and future prospects. Plant Dis. 102: 12-39.
15. Halleen F, Fourie PH, Crous PW (2006) A review of black foot disease of grapevine. Phytopathol. Mediterr. 45: 55-67.
16. Lim L, Mohd MH, Zakaria L (2019) Identification and pathogenicity of Diaporthe species associated with stem end rot of mango (Mangifera indica). Eur. J. Plant Pathol. 155: 687-696.
17. Mostert L, Halleen F, Fourie P, Crous PW (2006) A review of Phaeoacremonium species involved in Petri disease and esca of grapevines. Phytopathol. Mediterr. 45: 12-29.
18. Rawnsley B, Wicks TJ, Scott ES, Stummer BE (2004) Diaporthe perjuncta does not cause Phomopsis cane and leaf spot disease of grapevine in Australia. Plant Dis. 88: 1005-1010.
19. Reis P, Pierron M, Larignon P, Lecomte P, Abou-Mansour E, Farine S, Bertsch C, Jacques A, Trotel-Aziz P, Rego C, Fontaine F (2019) Vitis methods to understand and develop strategies for diagnosis and sustainable control of grapevine trunk diseases. Phytopathology 109: 916-931.
20. Srivastava P, Andersen PC, Marois JJ, Wright DL, Srivastava M, Harmon PF (2013) Effect of phenolic compounds on growth and ligninolytic enzyme production in Botryosphaeria isolates. Crop Prot. 43: 146-156.
21. Sosnowski M, Lardner R, Wicks T, Scott E (2005) A rapid metod of screening grapevine cultivars for susceptibility to Eutypa dieback. The Australian and New Zealand Grapegrower & Winemaker 2: 14-16.
22. Urbez-Torres JR (2011) The status of Botryosphaeriaceae species infecting grapevines. Phytopathol. Mediterr. 50: 5-45.