First report of root-knot nematode, Meloidogyne incognita on calendula in Turkey

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Journal of Nematology

Society of Nematologists

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First report of root-knot nematode, Meloidogyne incognita on calendula in Turkey

Hürkan Ataş / Gülsüm Uysal / Çiğdem Gözel / Tevfik Özalp / Uğur Gözel / Zübeyir Devran *

Keywords : Calendula officinalis, Identification, Meloidogyne incognita

Citation Information : Journal of Nematology. Volume 53, Pages 1-5, DOI: https://doi.org/10.21307/jofnem-2021-041

License : (CC-BY-4.0)

Received Date : 06-January-2021 / Published Online: 01-April-2021

ARTICLE

ABSTRACT

Calendula officinalis L. (Asteraceae) is a traditional medicinal plant called pot marigold or English marigold. In this study, galled roots of pot marigold were collected from Balıkesir province of Turkey and egg masses were picked up from the roots of each plant with fine forceps. DNA was then extracted from samples and analyzed by species-specific primers referring to the most common Meloidogyne spp. Our results showed that Meloidogyne incognita was found as the only species in all the samples taken. The determination of M. incognita on calendula was done for the first time in Turkey.

Graphical ABSTRACT

Plant parasitic nematodes, especially root-knot nematodes (Meloidogyne spp.), cause serious yield losses in vegetables, ornamental and medicinal plants, and horticultural crops around the world (Devran et al., 2017; Karssen et al., 2013; Pintea et al., 2003). The plantation areas of ornamental and medicinal plants in tropical and sub-tropical countries have significantly increased recently in order to be used in pharmaceutical, perfumery, cosmetic and food industries (Pandey, 2017). Calendula officinalis L. (Asteraceae) has been planted as a traditional medicinal plant and is medicinally used in the world (Muley et al., 2009). Its extracts possess a wide range of pharmacological effects (Pintea et al., 2003). However, ornamental and medicinal plants can be attacked by several soil borne pathogens. Russo et al. (2008) pointed out that C. officinalis was infested with Meloidogyne incognita (Kofoid and White, 1919), Chitwood, 1949 in Italy. In another study, Brito et al. (2010) reported root-knot nematodes infected ornamental plants including calendula in Florida. Histopathological changes induced by M. incognita were investigated in five ornamental plants including C. officinalis and it was found moderately susceptible based on the galling index (Siddiqui et al., 2014). Similarly, Alijani et al. (2015) showed that growth factors of C. officinalis significantly decreased with increasing the number of M. javanica (Treub, 1885) Chitwood, 1949. To the best of our knowledge, in Turkey, the infestation of root-knot nematodes on calendula have not been reported.

We surveyed on calendula plants grown in open fields of Balıkesir province of Turkey, we observed symptoms of wilting in some calendula plants. When examining the roots of these plants, we observed galls on the roots (Fig. 1). Egg masses on the roots of calendula were collected using forceps and were put into incubation for hatching of second stage juveniles were fixed in TAF fixative and permanent preparations were made according to Seinhorst’s (1959) method. Measurements of approximately 25 J2s were made according to Karssen (2002) under Leica DM1000 stereomicroscope (Table 1). Adult females were removed from the roots of the calendula plants with a needle and scalpel under binoculars. Perineal patterns of the extracted females were cut in 45% lactic acid and their preparations were made in glycerin (Hooper, 1986). Morphological identification was made by us according to Jepson (1987) and Karssen (2002) (Fig. 2). The overall morphology and morphometric measurements of this population appear to be similar to M. incognita (Eisenback and Triantaphyllou, 1991; Whitehead, 1968). For molecular identification of Meloidogyne sp., genomic, DNA was isolated from J2s using the High Pure PCR Template Preparation Kit (Roche). Subsequently, DNA was analyzed by species-specific primers referring to common root-knot nematode species M. incognita, M. javanica, M. arenaria (Neal, 1889) Chitwood, 1949, M. hapla (Chitwood, 1949), M. fallax (Karssen, 1996), and M. chitwoodi (Golden et al., 1980) (Devran et al., 2018; Randig et al., 2002; Wishart et al., 2002; Zijlstra et al., 2000). PCR reactions were carried out in a SimpliAmpTM Thermal Cycler (Applied Biosystems, CA, USA) according to Özalp et al. (2020). PCR products were run on a 2% agarose gel in 1X TAE buffer. Agarose gel was dyed with Xpert Green DNA Stain and viewed using the Gel iX Imager (Intas Science, Göttingen, Germany). M. incognita-specific inc-K14F/inc-K14R primers (Randig et al., 2002) and MincF1/MincR1 primer set (Devran et al., 2018) primers only produced an expected approximately 400 and 150 bp products, respectively, but other primers failed to amplify any products (Fig. 3).

Figure 1:

Galls caused by Meloidogyne incognita on the root of Calendula officinalis L.

10.21307_jofnem-2021-041-f001.jpg
Figure 2:

Meloidogyne incognita (Kofoid & White, 1919) from Calendula officinalis A: Anterior end region, B: Tail region, C-D: Perineal pattern (Scale bar: 20 µm).

10.21307_jofnem-2021-041-f002.jpg
Table 1.

Morphometric characters of second-stage juveniles (J2) of Meloidogyne incognita on Calendula officinalis.

10.21307_jofnem-2021-041-t001.jpg
Figure 3:

Amplified DNA of using Meloidogyne incognita-specific primers A: MincF1/MincR1 primer set, B: Inc-K14F/Inc-K14R primer set. M: DNA Ladder, Hibrigen 100 bp; BA1-BA5: Samples; S6: M. incognita (positive control); W: Water.

10.21307_jofnem-2021-041-f003.jpg

This is the first report identification of M. incognita on calendula in Turkey. It was also reported for the first time that plant parasitic nematodes on calendula in Turkey. The results demonstrated that calendula-growing areas in Balıkesir province were infested with M. incognita. These findings are important information for floriculturists and can be used to manage the damage caused by M. incognita.

References


  1. Alijani, Z. , Olia, M. , Sharifnabi, B. and Jaimand, K. 2015. Effect of different inoculum densities of root-knot nematode, Meloidogyne javanica on progress of disease and production of secondary compounds in pot marigold, Calendula officinalis , Iranian Journal of Plant Pathology 51:215–25.
  2. Brito, J. A. , Kaur, R. , Cetintas, R. , Stanley, J. D. , Mendes, M. L. , Powers, T. O. and Dickson, D. W. 2010. Meloidogyne spp. infecting ornamental plants in Florida. Nematropica 40:87–103.
  3. Chitwood, B. G. 1949. Root-knot nematodes Part 1. A revision of the genus Meloidogyne Goeldi, 1987. Proceedings of the helminthological Society of Washington 16:90–104.
  4. Devran, Z. , Mıstanoğlu, İ. and Özalp, T. 2017. Occurrence of mixed populations of root-knot nematodes in vegetable greenhouses in Turkey, as determined by PCR screening. Journal of Plant Diseases and Protection 124:617–30.
  5. Devran, Z. , Polat, İ. , Mıstanoğlu, İ. and Baysal, Ö. 2018. A novel multiplex PCR tool for simultaneous detection of three root-knot nematodes. Australasian Plant Pathology 47:389–92.
  6. Eisenback, J. D. , Triantaphyllou, H. H. 1991. Root-knot nematodes: Meloidogyne species and races. Manual of Agricultural Nematology, 1:191–274.
  7. Golden, A. M. , O’Bannon, H. , Santo, G. S. and Finley, A. M. 1980. Description and SEM observations of Meloidogyne chitwoodi n. sp. (Meloidogynidae), a root-knot nematode on potato in the Pacific Northwest. Journal of Nematology 12:319–27.
  8. Hooper, D. J. 1986. Extraction of free-living stages from soil, In Southey, J. F. (Ed.), Laboratory Methods for Work with Plant Soil Nematodes, London: Her Majesty’s Stationary Office, pp. 5–30.
  9. Jepson, S. B. 1987. Identification of Root-knot Nematodes Meloidogyne species CABI Wallingford, p. 265.
  10. Karssen, G. 1996. Description of Meloidogyne fallax n. sp. (Nematoda: Heteroderidae), a root-knot nematode from the Netherlands. Fundamental and Applied Nematology 19:593–600.
  11. Karssen, G. 2002. The plant parasitic nematode genus Meloidogyne Goeldi, 1892 (Tylenchida) in Europe. Leiden: Brill Academic Publishers.
  12. Karssen, G. , Wesemael, W. and Moens, M. 2013. “Root-knot nematodes”. In Perry, R. N. and Moens, M. (Eds), Plant Nematology, 2nd ed., Wallingford: CAB International, pp. 59–90.
  13. Kofoid, C. A. and White, A. W. 1919. A new nematode infection of man. Journal of the American Medical Association 72:567–569.
  14. Muley, B. , Khadabadi, S. and Banarase, N. 2009. Phytochemical constituents and pharmacological activities of Calendula officinalis Linn (Asteraceae): a review. Tropical Journal of Pharmaceutical Research 8:455–65.
  15. Neal, J. C. 1889. The root-knot disease of the peach, orange and other plants in Florida due to the work of Anguillula, Bulletin US Bureau of Entomology.
  16. Özalp, T. , Könül, G. , Ayyıldız, Ö. , Tülek, A. and Devran Z. 2020. First report of root-knot nematode Meloidogyne arenaria, on lavender in Turkey. Journal of Nematology 52:1–3.
  17. Pandey, R. 2017. Diseases of medicinal and aromatic plants: insights in nematode biomanagement. Indian Phytopathology 70:12–21.
  18. Pintea, A. , Bele, C. , Andrei, S. and Socaciu, C. 2003. HPLC analysis of carotenoids in four varieties of Calendula officinalis L. flowers. Acta Biologica Szegediensis 47:37–40.
  19. Randig, O. , Bongiovanni, M. , Carneiro, R. M. and Castagnone-Sereno, P. 2002. Genetic diversity of root knot nematodes from Brazil and development of SCAR marker specific for the coffee damaging species. Genome 45:862–70.
  20. Russo, G. , Soppelsa, O. and D’Errico, G. 2008. Infestation of root-knot nematodes (Meloidogyne spp.) on two flower plants of new introduction: Calendula officinalis L. and Paeonia lactiflora Pall. REDIA-J Xoology 91:163–66.
  21. Seinhorst, J. W. 1959. A rapid method for the transfer of nematodes from fixative to anhydrous glycerin. Nematologica, 4:67–69.
  22. Siddiqui, Y. , Ali, A. and Naidu, Y. 2014. Histopathological changes induced by Meloidogyne incognita in some ornamental plants. Crop Protection 65:216–20.
  23. Treub, M. 1885. Onderzoekingen over sereh-ziek suikerriet gedaan in s’Lands Plantentuin te Buitenzorg. Meded. uit ‘s lands plantentuin Batavia 2:1–39.
  24. Whitehead, A. G. 1968. Taxonomy of Meloidogyne (Nematodea: Heteroderidae) with descriptions of four new species. Transactions of Zoological Society of London, 31:263–401.
  25. Wishart, J. , Phillips, M. S. and Blok, V. C. 2002. Ribosomal Intergenic Spacer: a Polymerase chain reaction diagnostic for Meloidogyne chitwoodi, M. fallax, and M. hapla . Nematology 92:884–92.
  26. Zijlstra, C. , Donkers-Venne, D. T. H. M. and Fargette, M. 2000. Identification of Meloidogyne incognita, M. javanica and M. arenaria using sequence characterized amplified region (SCAR) based PCR assays. Nematology 2:847–53.
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FIGURES & TABLES

Figure 1:

Galls caused by Meloidogyne incognita on the root of Calendula officinalis L.

Full Size   |   Slide (.pptx)

Figure 2:

Meloidogyne incognita (Kofoid & White, 1919) from Calendula officinalis A: Anterior end region, B: Tail region, C-D: Perineal pattern (Scale bar: 20 µm).

Full Size   |   Slide (.pptx)

Figure 3:

Amplified DNA of using Meloidogyne incognita-specific primers A: MincF1/MincR1 primer set, B: Inc-K14F/Inc-K14R primer set. M: DNA Ladder, Hibrigen 100 bp; BA1-BA5: Samples; S6: M. incognita (positive control); W: Water.

Full Size   |   Slide (.pptx)

REFERENCES

  1. Alijani, Z. , Olia, M. , Sharifnabi, B. and Jaimand, K. 2015. Effect of different inoculum densities of root-knot nematode, Meloidogyne javanica on progress of disease and production of secondary compounds in pot marigold, Calendula officinalis , Iranian Journal of Plant Pathology 51:215–25.
  2. Brito, J. A. , Kaur, R. , Cetintas, R. , Stanley, J. D. , Mendes, M. L. , Powers, T. O. and Dickson, D. W. 2010. Meloidogyne spp. infecting ornamental plants in Florida. Nematropica 40:87–103.
  3. Chitwood, B. G. 1949. Root-knot nematodes Part 1. A revision of the genus Meloidogyne Goeldi, 1987. Proceedings of the helminthological Society of Washington 16:90–104.
  4. Devran, Z. , Mıstanoğlu, İ. and Özalp, T. 2017. Occurrence of mixed populations of root-knot nematodes in vegetable greenhouses in Turkey, as determined by PCR screening. Journal of Plant Diseases and Protection 124:617–30.
  5. Devran, Z. , Polat, İ. , Mıstanoğlu, İ. and Baysal, Ö. 2018. A novel multiplex PCR tool for simultaneous detection of three root-knot nematodes. Australasian Plant Pathology 47:389–92.
  6. Eisenback, J. D. , Triantaphyllou, H. H. 1991. Root-knot nematodes: Meloidogyne species and races. Manual of Agricultural Nematology, 1:191–274.
  7. Golden, A. M. , O’Bannon, H. , Santo, G. S. and Finley, A. M. 1980. Description and SEM observations of Meloidogyne chitwoodi n. sp. (Meloidogynidae), a root-knot nematode on potato in the Pacific Northwest. Journal of Nematology 12:319–27.
  8. Hooper, D. J. 1986. Extraction of free-living stages from soil, In Southey, J. F. (Ed.), Laboratory Methods for Work with Plant Soil Nematodes, London: Her Majesty’s Stationary Office, pp. 5–30.
  9. Jepson, S. B. 1987. Identification of Root-knot Nematodes Meloidogyne species CABI Wallingford, p. 265.
  10. Karssen, G. 1996. Description of Meloidogyne fallax n. sp. (Nematoda: Heteroderidae), a root-knot nematode from the Netherlands. Fundamental and Applied Nematology 19:593–600.
  11. Karssen, G. 2002. The plant parasitic nematode genus Meloidogyne Goeldi, 1892 (Tylenchida) in Europe. Leiden: Brill Academic Publishers.
  12. Karssen, G. , Wesemael, W. and Moens, M. 2013. “Root-knot nematodes”. In Perry, R. N. and Moens, M. (Eds), Plant Nematology, 2nd ed., Wallingford: CAB International, pp. 59–90.
  13. Kofoid, C. A. and White, A. W. 1919. A new nematode infection of man. Journal of the American Medical Association 72:567–569.
  14. Muley, B. , Khadabadi, S. and Banarase, N. 2009. Phytochemical constituents and pharmacological activities of Calendula officinalis Linn (Asteraceae): a review. Tropical Journal of Pharmaceutical Research 8:455–65.
  15. Neal, J. C. 1889. The root-knot disease of the peach, orange and other plants in Florida due to the work of Anguillula, Bulletin US Bureau of Entomology.
  16. Özalp, T. , Könül, G. , Ayyıldız, Ö. , Tülek, A. and Devran Z. 2020. First report of root-knot nematode Meloidogyne arenaria, on lavender in Turkey. Journal of Nematology 52:1–3.
  17. Pandey, R. 2017. Diseases of medicinal and aromatic plants: insights in nematode biomanagement. Indian Phytopathology 70:12–21.
  18. Pintea, A. , Bele, C. , Andrei, S. and Socaciu, C. 2003. HPLC analysis of carotenoids in four varieties of Calendula officinalis L. flowers. Acta Biologica Szegediensis 47:37–40.
  19. Randig, O. , Bongiovanni, M. , Carneiro, R. M. and Castagnone-Sereno, P. 2002. Genetic diversity of root knot nematodes from Brazil and development of SCAR marker specific for the coffee damaging species. Genome 45:862–70.
  20. Russo, G. , Soppelsa, O. and D’Errico, G. 2008. Infestation of root-knot nematodes (Meloidogyne spp.) on two flower plants of new introduction: Calendula officinalis L. and Paeonia lactiflora Pall. REDIA-J Xoology 91:163–66.
  21. Seinhorst, J. W. 1959. A rapid method for the transfer of nematodes from fixative to anhydrous glycerin. Nematologica, 4:67–69.
  22. Siddiqui, Y. , Ali, A. and Naidu, Y. 2014. Histopathological changes induced by Meloidogyne incognita in some ornamental plants. Crop Protection 65:216–20.
  23. Treub, M. 1885. Onderzoekingen over sereh-ziek suikerriet gedaan in s’Lands Plantentuin te Buitenzorg. Meded. uit ‘s lands plantentuin Batavia 2:1–39.
  24. Whitehead, A. G. 1968. Taxonomy of Meloidogyne (Nematodea: Heteroderidae) with descriptions of four new species. Transactions of Zoological Society of London, 31:263–401.
  25. Wishart, J. , Phillips, M. S. and Blok, V. C. 2002. Ribosomal Intergenic Spacer: a Polymerase chain reaction diagnostic for Meloidogyne chitwoodi, M. fallax, and M. hapla . Nematology 92:884–92.
  26. Zijlstra, C. , Donkers-Venne, D. T. H. M. and Fargette, M. 2000. Identification of Meloidogyne incognita, M. javanica and M. arenaria using sequence characterized amplified region (SCAR) based PCR assays. Nematology 2:847–53.

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