Biber hafif benek virüs’üne (PMMoV) karşı L4 dayanıklılık durumunun taranması ve moleküler yöntemlerle karakterizasyonu
Öz
Biber hafif benek virüsü (Pepper mild mottle virus- PMMoV) Tobamovirüsler içerisinde yer alan dünya genelinde biber üretim
alanlarında en sık karşılaşılan virüslerden biridir. Bu virüs çoğu zaman hasat
dönemine kadar gizli kalabilmekte ve en büyük zararlarını hasat döneminde
meyveler üzerinde meydana getirebilmektedir. Bölgemizde son yıllarda biber
üretim alanlarında sıklıkla karşılaşılan bir problem olarak karşımıza
çıkmaktadır. Dünyanın farklı bölgelerinde L4 geni vasıtasıyla sağlanan
dayanıklılığı kıran virüs izolatının rapor edilmesi, ülkemizde de PMMoV’nin epidemi
durumunun araştırılması gerekliliğini ortaya koymuştur. L4 dayanıklılığının etkinliğini
belirleyebilmek için bölgemizden biber üretim alanlarında PMMoV ile şüpheli
örnekler toplanmıştır. RT-PCR tekniği ile doğrulanan bu örnek PMMoV-Kum olarak
kodlanmıştır ve Gen bankasına kaydı yapılarak erişim numarası (MK806437)
alınmıştır. PMMoV-Kum izolatı 1 hassas (B1) ve 5 dayanıklı (L4B1, L4B2, L4B3, L4B4,
L4B5) bitkilerine mekanik inokulasyon yöntemi ile bulaştırılmıştır. Yapılan
fenotipik gözlemler neticesinde PMMoV-Kum izolatının L4 genine bağlı
dayanıklılığı kıramadığı biyolojik testler ile doğrulanmıştır. Dayanıklı olarak
seçilen bitkiler üzerinde herhangi bir PMMoV simptomunun oluşmaması üzerine en
başarılı çalışan L4 markırlarını belirlemeye yönelik primerlerin belirlenmesi
tez çalışmasının seyrini belirlemiş ve en başarılı markırın AP-7/AP-8 ve
P118/119primer çiftleri olduğu tespit edilmiştir.
Anahtar Kelimeler
L4 geni Dayanıklılığın kırılması Biber hafif benek virüsü Biber PMMoV
Destekleyen Kurum
Akdeniz Üniversitesi Bitki Koruma Bölümü
Proje Numarası
FYL-2018-3638
Teşekkür
Bu Tez Akdeniz Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından desteklenmiştir. Teşekkür Ederiz.
Kaynakça
- Antignus Y, Lachman O, Pearlsman M, Maslenin L, Rosner A (2008) A New Pathotype of Pepper mild mottle virus (PMMoV) Overcomes the L4 Resistance Genotype of Pepper Cultivars. Plant Disease 92(7): 1033–1037.
- Berzal-Herranz A, de la Cruz A, Tenllado F, Díaz-Ruíz JR, López L, Sanz AI, Vaquero C, Serra MT, García-Luque I (1995) The Capsicum L3 gene-mediated resistance against the Tobamoviruses is elicited by the coat protein. Virology 209: 498-505.
- Boukema IW (1980) Allelism of genes controlling resistance to TMV in Capsicum L. Euphytica 29: 433–439.
- Boukema IW (1983) Research on the location of the gene for resistance to TMV in Capsicum chacoense Hunz. and male sterility in progenies from the cross C. chacoense×C. annuum L. Proceedings Vth Meeting Capsicum and Eggplant Working Group of Eucarpia, Bulgaria, p. 84-87.
- Boukema IW (1984) Resistance to TMV in Capsicum chacoense Hunz. is governed by an allele of the L-locus. Capsicum Newsletter 3: 47-48.
- Choi GS, Choi SK, Cho JD, Cho IS (2013) A pathotype of Pepper Mild Mottle Virus causing necrotic spot symptoms in paprika fruit. Research in Plant Disease 19: 124-127.
- Choi GS, Choi SK, Cho IS, Kwon SJ (2014) Resistance screening to Pepper Mild Mottle iVrus pathotypes in paprika cultivars. Research in Plant Disease 20: 299-302.
- Csillery G, Tobias I, Rusko J (1983) A New Pepper Strain of Tomato Mosaic Virus. Acta Phytopathologica Academiae Scientiarum Hungaricae 18: 195-200.
- Çağlar BK, Fidan H, Elbeaino T (2012) Detection and Molecular Characterization of Pepper Mild Mottle Virus from Turkey. Journal of Phytopathology 161(6): 434-438.
- FAOSTAT (2018) Statistical database. http://www. fao.org/faostat/en/#data/QC. Accessed 11 May 2018.
- Fidan H, Sarı N (2019) Molecular Characterization of Resistance-Breaking Tomato Spotted Wilt Virus (TSWV) Isolate Medium Segment in Tomato. Applıed Ecology and Environmental Research 17(2): 5321-5339.
- Genda Y, Kanda A, Hamada H, Sato K, Ohnishi J, Tsuda S (2007) Two amino acid substitutions in the coat protein of Pepper mild mottle virus are responsible for overcoming the L4 gene-mediated resistance in Capsicum spp. Phytopathology 97: 787-793.
- Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Oxford University Prees, Nucleic Acids Syposium Series No.41: 95-98.
- Janzac B, Fabre MF, Palloıx A, Moury B (2009) Phenotype and spectrum of action of the Pvr4 resistance in pepper against potyviruses, and selection of virulent variants. Plant Pathology 58: 443-449.
- Kim HJ, Han JH, Yoo JH, Cho HJ, Kim BD (2008) Development of a sequence characteristic amplified region marker linked to the L4 locus conferring broad spectrum resistance to tobamoviruses in pepper plants. Molecules and Cells 25: 205-210.
- Kumar S, Udaya AC, Nayaka SC, Lund OS, Prakas HS (2011) Detection of Tobacco mosaic virus and Tomato mosaic virus in pepper and tomato by multiplex RT-PCR. Letter in applied Microbiology 359-363.
- Lefebvre V, Pflieger S, Thabuis A, Caranta C, Blattes A, Chauvet JC, Daubeze AM, Palloix A (2002) Towards the saturation of the pepper linkage map by alignment of three intraspecific maps including known-function genes. Genome 45(5): 839-854.
- Matsunaga HT, Saito M, Hırai T, Yoshıda T (2003) DNA markers linked to pepper mild mottle virus (PMMoV) resistant locus (L4) in Capsicum. Japanese Society for Horticultural Science 72: 218-220.
- Milosevic D, Stankovic I, Bulajic A, Nikolic Z, Ignjatov M, Krstic B (2012) Molecular characterization of Pepper mild mottle virus in Serbia. Genetıka 47(2): 651-663.
- Pernezny K, Roberts PD, Murphy JF, Goldberg NP (2003) Compendium of Pepper Diseases. St. Paul, Minnedota: The American Phytopathological Society, p. 73.
- Rast ATB (1988) Pepper tobamoviruses and pathotypes used in resistance breeding. Capsicum Newsletter 7: 20-23.
- Sawada H, Takeuchi S, Matsumoto K, Hamada H, Kiba A, Matsumoto M, Watanabe Y, Suzuki K, Hikichi Y (2005) A new Tobamovirus-resistance gene, Hk, in Capsicum annuum. Japanese Society for Horticultural Science 74: 289-294.
- Scholthof KBGS, Adkıns H, Czosnek P, Palukaitis E, Jacquot T, Hohn B, Hoh K, Saunders T, Candresse P, Ahlquist C, Foster GD (2011) Top 10 plant viruses in molecular plant pathology. Molecular Plant Pathology 12: 938-954.
- Tomita R, Ken-Taro S, Hiroyuki M, Sakamoto M, Murai J, Kiba A, Hikichi Y, Suzuki K, Kobayashi K (2011) Genetic basis for the hierarchical interaction between Tobamovirus spp. and L resistance gene alleles from different pepper species. Molecular Plant-Microbe Interactions 24: 108-117.
- TUİK (2018) Statistical database. http://www.tuik.gov.tr/ PreTablo.do?alt_id=1001. Accessed 01 May 2018.
- Van Duin PJW (1998) Xth EUCARPIA Meeting on Genetics and Breeding of Capsicum & Eggplant, Avignon, France.
- Velasco L, Janssen D, Ruiz-Garcia L, Segundo E, Cuadrado IM (2002) The complete nucleotide sequence and development of a diferential detection assay for a pepper mild mottle virus (PMMoV) isolate that overcomes L3 resistance in pepper. Journal of Virological Methods 106(1): 135–140.
- Wetter C, Conti M, Altschuh D, Tabillion R, Van Regenmortel MHV (1984) Pepper Mild Mottle Virus, a Tobamovirus infecting pepper cultivars in Sicily. Phytopathology 74(4): 405-410.
- Yang HB, Lıu WY, Kang WH, Jahn M, Kang BC (2009) Development of SNP markers linked to the L locus in Capsicum spp. by a comparative genetic analysis. In Molecular Breeding 24(4): 433-446.
Screening of L4 resistance status to pepper mild mottle virus (PMMoV) and characterization by molecular methods
Öz
Pepper mild mottle virus (PMMoV) is one of the
most common Tobamoviruses can be found in worldwide. This virus has
potential to cause economic losses and has kept secret infections until harvest
time causing great damages on pepper fruits at this period. During 2016-2019
production periods, it has been frequently seen in pepper grown areas. Several
isolates reported from different regions of the world regarding resistance with
L4 gene fights against this virus disease. In this study, we aimed to
investigate how the PMMoV increased and understood its incidence pepper grown
areas in Turkey. In order to determine the effectiveness of L4 resistance,
PMMoV and other suspected pepper samples were isolated. The collected samples
were studied by RT-PCR technique using PMMoV-Kum isolate and its gene bank’s
access number (MK806437) was registered. A sensitive (B1) and 5 resistant
(L4B1, L4B2, L4B3, L4B4, L4B5) plants were mechanically inoculated with the
PMMoV-Kum isolate and their phenotypic observations were recorded. The
PMMoV-Kum isolate was confirmed as avirulent in biological tests, the PMMoV
could not overcome L4 gene mediated resistance. Furthermore, a
successful primer set has determined as AP-7 / AP-8 in L4 gene mediated
resistance against occurrence of any PMMoV symptoms on the tested plants.
Determining new PMMoV isolates which break the L4 gene mediated resistance, phylogenetic analyzes was performed to
determine its place among the world’s isolates.
Anahtar Kelimeler
L4 gene Resistance Breaking Pepper Pepper mild mottle virus PMMoV
Proje Numarası
FYL-2018-3638
Kaynakça
- Antignus Y, Lachman O, Pearlsman M, Maslenin L, Rosner A (2008) A New Pathotype of Pepper mild mottle virus (PMMoV) Overcomes the L4 Resistance Genotype of Pepper Cultivars. Plant Disease 92(7): 1033–1037.
- Berzal-Herranz A, de la Cruz A, Tenllado F, Díaz-Ruíz JR, López L, Sanz AI, Vaquero C, Serra MT, García-Luque I (1995) The Capsicum L3 gene-mediated resistance against the Tobamoviruses is elicited by the coat protein. Virology 209: 498-505.
- Boukema IW (1980) Allelism of genes controlling resistance to TMV in Capsicum L. Euphytica 29: 433–439.
- Boukema IW (1983) Research on the location of the gene for resistance to TMV in Capsicum chacoense Hunz. and male sterility in progenies from the cross C. chacoense×C. annuum L. Proceedings Vth Meeting Capsicum and Eggplant Working Group of Eucarpia, Bulgaria, p. 84-87.
- Boukema IW (1984) Resistance to TMV in Capsicum chacoense Hunz. is governed by an allele of the L-locus. Capsicum Newsletter 3: 47-48.
- Choi GS, Choi SK, Cho JD, Cho IS (2013) A pathotype of Pepper Mild Mottle Virus causing necrotic spot symptoms in paprika fruit. Research in Plant Disease 19: 124-127.
- Choi GS, Choi SK, Cho IS, Kwon SJ (2014) Resistance screening to Pepper Mild Mottle iVrus pathotypes in paprika cultivars. Research in Plant Disease 20: 299-302.
- Csillery G, Tobias I, Rusko J (1983) A New Pepper Strain of Tomato Mosaic Virus. Acta Phytopathologica Academiae Scientiarum Hungaricae 18: 195-200.
- Çağlar BK, Fidan H, Elbeaino T (2012) Detection and Molecular Characterization of Pepper Mild Mottle Virus from Turkey. Journal of Phytopathology 161(6): 434-438.
- FAOSTAT (2018) Statistical database. http://www. fao.org/faostat/en/#data/QC. Accessed 11 May 2018.
- Fidan H, Sarı N (2019) Molecular Characterization of Resistance-Breaking Tomato Spotted Wilt Virus (TSWV) Isolate Medium Segment in Tomato. Applıed Ecology and Environmental Research 17(2): 5321-5339.
- Genda Y, Kanda A, Hamada H, Sato K, Ohnishi J, Tsuda S (2007) Two amino acid substitutions in the coat protein of Pepper mild mottle virus are responsible for overcoming the L4 gene-mediated resistance in Capsicum spp. Phytopathology 97: 787-793.
- Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Oxford University Prees, Nucleic Acids Syposium Series No.41: 95-98.
- Janzac B, Fabre MF, Palloıx A, Moury B (2009) Phenotype and spectrum of action of the Pvr4 resistance in pepper against potyviruses, and selection of virulent variants. Plant Pathology 58: 443-449.
- Kim HJ, Han JH, Yoo JH, Cho HJ, Kim BD (2008) Development of a sequence characteristic amplified region marker linked to the L4 locus conferring broad spectrum resistance to tobamoviruses in pepper plants. Molecules and Cells 25: 205-210.
- Kumar S, Udaya AC, Nayaka SC, Lund OS, Prakas HS (2011) Detection of Tobacco mosaic virus and Tomato mosaic virus in pepper and tomato by multiplex RT-PCR. Letter in applied Microbiology 359-363.
- Lefebvre V, Pflieger S, Thabuis A, Caranta C, Blattes A, Chauvet JC, Daubeze AM, Palloix A (2002) Towards the saturation of the pepper linkage map by alignment of three intraspecific maps including known-function genes. Genome 45(5): 839-854.
- Matsunaga HT, Saito M, Hırai T, Yoshıda T (2003) DNA markers linked to pepper mild mottle virus (PMMoV) resistant locus (L4) in Capsicum. Japanese Society for Horticultural Science 72: 218-220.
- Milosevic D, Stankovic I, Bulajic A, Nikolic Z, Ignjatov M, Krstic B (2012) Molecular characterization of Pepper mild mottle virus in Serbia. Genetıka 47(2): 651-663.
- Pernezny K, Roberts PD, Murphy JF, Goldberg NP (2003) Compendium of Pepper Diseases. St. Paul, Minnedota: The American Phytopathological Society, p. 73.
- Rast ATB (1988) Pepper tobamoviruses and pathotypes used in resistance breeding. Capsicum Newsletter 7: 20-23.
- Sawada H, Takeuchi S, Matsumoto K, Hamada H, Kiba A, Matsumoto M, Watanabe Y, Suzuki K, Hikichi Y (2005) A new Tobamovirus-resistance gene, Hk, in Capsicum annuum. Japanese Society for Horticultural Science 74: 289-294.
- Scholthof KBGS, Adkıns H, Czosnek P, Palukaitis E, Jacquot T, Hohn B, Hoh K, Saunders T, Candresse P, Ahlquist C, Foster GD (2011) Top 10 plant viruses in molecular plant pathology. Molecular Plant Pathology 12: 938-954.
- Tomita R, Ken-Taro S, Hiroyuki M, Sakamoto M, Murai J, Kiba A, Hikichi Y, Suzuki K, Kobayashi K (2011) Genetic basis for the hierarchical interaction between Tobamovirus spp. and L resistance gene alleles from different pepper species. Molecular Plant-Microbe Interactions 24: 108-117.
- TUİK (2018) Statistical database. http://www.tuik.gov.tr/ PreTablo.do?alt_id=1001. Accessed 01 May 2018.
- Van Duin PJW (1998) Xth EUCARPIA Meeting on Genetics and Breeding of Capsicum & Eggplant, Avignon, France.
- Velasco L, Janssen D, Ruiz-Garcia L, Segundo E, Cuadrado IM (2002) The complete nucleotide sequence and development of a diferential detection assay for a pepper mild mottle virus (PMMoV) isolate that overcomes L3 resistance in pepper. Journal of Virological Methods 106(1): 135–140.
- Wetter C, Conti M, Altschuh D, Tabillion R, Van Regenmortel MHV (1984) Pepper Mild Mottle Virus, a Tobamovirus infecting pepper cultivars in Sicily. Phytopathology 74(4): 405-410.
- Yang HB, Lıu WY, Kang WH, Jahn M, Kang BC (2009) Development of SNP markers linked to the L locus in Capsicum spp. by a comparative genetic analysis. In Molecular Breeding 24(4): 433-446.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Ziraat Mühendisliği |
| Bölüm | Makaleler |
| Yazarlar | |
| Proje Numarası | FYL-2018-3638 |
| Yayımlanma Tarihi | 1 Aralık 2019 |
| Gönderilme Tarihi | 19 Temmuz 2019 |
| Yayımlandığı Sayı | Yıl 2019 Cilt: 32 Sayı: 3 |
Kaynak Göster
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