First report of Meloidogyne javanica infecting Zinnia elegans in Ceará State, Brazil

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Journal of Nematology

Society of Nematologists

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ISSN: 0022-300X
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First report of Meloidogyne javanica infecting Zinnia elegans in Ceará State, Brazil

Francisco Jorge Carlos Souza Junior * / Mayara Castro Assunção

Keywords : Meloidogyne javanica , Molecular biology, Morphology, Ribosome markers, Root-knot nematodes, Zinnia elegans

Citation Information : Journal of Nematology. Volume 52, Pages 1-4, DOI: https://doi.org/10.21307/jofnem-2020-066

License : (CC-BY-4.0)

Received Date : 25-April-2020 / Published Online: 28-July-2020

ARTICLE

ABSTRACT

Roots of Zinnia elegans exhibiting galls were observed and collected in the city of Pacoti, CE, Brazil. Through the morphological characterization and molecular analysis of the ITS and 28S rDNA regions, the causal agent was identified as a nematode of the genus Meloidogyne species M. javanica. This is the first detection of M. javanica in Zinnia elegans in the state of Ceará in Brazil.

Graphical ABSTRACT

Zinnia (Zinnia elegans Jacq.), a herbaceous plant of the Asteraceae family, is among the species use in Brazil for flower production. This species, originally from Mexico, has a broad diversity of flower colors, a wide variety of petal shapes, and the possibility of being cultivated throughout the year (Torres, 1963; Stimart, 1987).

The Zinnia genus comprises 17 species and in Brazil it is known by common names as captain, young, and old, cinnamon sticks or zinnia, being grown in regions of mild temperature and in tropical areas. The plant has an erect shape, oval leaves, and colorful floral rays of various forms (Torres, 1963; Ahmad et al., 2015).

The genus Meloidogyne (Goeldi, 1887), also known as gall nematode, is widespread in all regions of Brazil, associated with agricultural crops. Alternative hosts, such as ornamental plants, maintain the inoculum and increase its populations, compromising the development of plants and affecting the quality of products and resulting in losses in productivity (Moens et al., 2009).

In October 2019, samples of roots of Zinnia elegans with galls caused by root-knot nematode were sampled in vegetable gardens in the city of Pacoti (4° 13′ 45″S; 38° 55′ 26″ W), in the state of Ceará, Brazil. Nematodes in the roots were processed for extraction using the protocol given by Coolen and D’Herde (1972). For the morphological characterization of the species, 20 females and juveniles of second stage were measured and perineal pattern from 20 females were prepared according to Taylor and Netscher (1974). The determination of the esterase profile was made according to Carneiro and Almeida (2001), using 20 female.

For molecular identification, the D2 to D3 region of 28S rDNA segment was amplified and sequenced using the primers D2A (5′-ACAAGTACCGTGAGGGAAAGTTG-3′) and D3B (5′-TCGGAAGGAACCAGCTACTA-3′) (De Ley et al., 1999) and ITS primers with VRAIN2F (5′-CTTTGTACACACCGCCCGTCGCT-3′) and VRAIN2R (5′-TTTCACTCGCCGTTACTAAGGGAATC-3′) (Vrain et al., 1992).

The consensus sequences were assembled from the forward and reverse sequences, using the Staden package (Staden et al., 2000). All consensus sequences obtained were used to compare with the NCBI nucleotide database, based on the research using the BLAST algorithm. Several sequence alignments for each individual gene were generated with the online version of MAFFT version 7 with the iterative refinement method L-INS-i (Katoh and Toh, 2008; Katoh, 2013). Phylogenetic analysis was performed using the maximum likelihood (ML) methods for individual genes, performed via RAxML-HCP2 v.8.2.8 (Stamatakis, 2014) implemented in CIPRES Portal v.2.0 (https://www.phylo.org/portal2/home.action) with 1,000 repetitions. For the ML analysis, a GTR-GAMMA model of evolution was used.

In females (n = 20), the body length was 437.3 ± 34.1 (427.1-514.3) μm; the stylus measured 12.4 ± 0.4 (11.4-14.1) μm in length; the DGO was 3.0 ± 0.6 (2.8-4.5) μm. For second-stage juveniles (n = 20), the body length was 334.0 ± 24.3 (327.0-417.8) μm; stylus length 11.0 ± 1.4 (10.2-12.0) μm; DGO equal to 2.6 ± 0.5 (2.0-2.8) μm; and tail measurements c = 9.1 ± 0.8 (7.4-10.2) μm and c′ = 5.0 ± 0.6 (4,7-5.7) μm, showing tapering in the terminal region (Mai and Mullin, 1996). The perineal patterns of the females were rounded with the low dorsal arch, smooth streaks, and parallel lateral lines highlighted on both sides (Fig. 1A). The polymorphisms of the esterase bands by electrophoresis revealed the phenotype J3 (Rm = 1.00, 1.25, and 1.40) typical of M. javanica (Fig. 1B).

Figure 1:

(A) Perineal pattern of Meloidogyne javanica, (B) Esterase phenotypes of M. javanica detected in Zinnia elegans (J3: M. javanica; J3*: M. javanica reference isolate).

10.21307_jofnem-2020-066-f001.jpg

The sequences submitted to GenBank ITS: MT337435 and D2 to D3 28S: MT341299 of the studied rDNA regions, showed 98 to 99% identity with sequences of M. javanica isolates from Brazil, China, and India. Phylogenetic analyzes of these sequences, using ML, placed the Meloidogyne population (CN0027) isolated from Zinnia elegans in a clade with the M. javanica sequences from GenBank, confirming it as M. javanica (Fig. 2).

Figure 2:

Phylogenetic relationship of Meloidogyne javanica from Ceará, Brazil, based on alignment of the sequence of the (A) D2/D3 expansion fragments of 28S rDNA and (B) sequence of ITS. The phylogenetic tree was estimated by maximum likelihood. Pratylenchus scribneri and P. zeae was used an outgroup.

10.21307_jofnem-2020-066-f002.jpg

In 2014, M. javanica was reported associated with Z. elegans in Iraq (Ali et al., 2014) and, in Brazil in 2017, the species M. incognita (Moreira et al., 2017) and M. hapla (Freire and Mosca, 2009) were described in Z. elegans. This nematode has economic importance in the production areas, with wide geographical distribution and occupying the first position in the world ranking among the nematodes harmful to agricultural crops (Jones et al., 2013)

However, this is the first record of M. javanica parasitizing roots of Z. elegans in Brazil, and thus, this work updates the information on the occurrence of this species in Z. elegans, indicating this plant as a potential host for this nematode in the state of Ceará.

References


  1. Ahmad, I., Whipker, B. E. and Dole, J. M. 2015. Paclobutrazol or ancymidol effects on postharvest performance of potted ornamental plants and plugs. HortScience 50:1370–1374.
  2. Ali, H. H., Fatah, K. M. and Ahmed, A. I. 2014. First record of nematode root knot on Zennia elegans caused by Meloidogyne javanica plant in Erbil Governorate, Kurdistan Region, Iraq. Arab Journal of Plant Protection 32:276–277.
  3. Carneiro, R. M. D. G. and Almeida, M. R. A. 2001. Electrophoresis technique used in the study of root-knot nematode enzymes for species identification. Nematologia Brasileira 25:555–560.
  4. Coolen, W. A. and D’Herde, C. J. 1972. A method for the quantitative extraction of nematodes from plant tissue State Agricultural and Entomology Research Station, Ghent.
  5. De Ley, P., Felix, M. A., Frisse, L. M., Nadler, S. A., Sternberg, P. W. and Thomas, W. K. 1999. Molecular and morphological characterization of two reproductively isolated species with mirror-image anatomy (Nematoda: Cephalobidae). Nematology 1:591–612.
  6. Freire, F. D. C. O. and Mosca, J. L. 2009. Patógenos associados a doenças de plantas ornamentais no Estado do Ceará. Ornamental Horticulture 15:83–89.
  7. Goeldi, E. A. 1887. Relatório sobre a moléstia do cafeeiro na provincial da Rio de Janeiro. Archivos do Museu Nacional do Rio de Janeiro 8:7–123.
  8. Jones, J. T., Haegeman, A., Danchin, E. G. J., Gaur, H. S., Helder, J., Jones, M. G. K., Kikuchi, T., Manzanilla-López, R., Palomares-Rius, J. E., Wesemael, W. M. L. and Perry, R. N. 2013. Top 10 plant-parasitic nematodes in molecular plant pathology. Molecular Plant Pathology 14:946–961.
    [PUBMED]
  9. Katoh, K. and Toh, H. 2008. Recent developments in the MAFFT multiple sequence alignment program. Briefings in Bioinformatics 9:286–298.
    [PUBMED]
  10. Katoh, S. 2013. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30:772–780.
    [PUBMED]
  11. Mai, W. F. and Mullin, P. G. 1996. Plant parasitic nematodes: a pictorial key to genera, New York, NY: Cornell University Press.
  12. Moens, M., Perry, R. N. and Starr, J. L. 2009. “ Meloidogyne species – a diverse group of novel and important plant parasites”, In Perry, R. N., Moens, M. and Starr, J. L. (Eds), Root-knot nematodes, Wallingford: CABI, 1–18.
  13. Moreira, F. J. C., Santos, C. D. G., Silva, G. S. and Innecco, R. 2017. Hospedabilidade de plantas ornamentais e medicinais ao nematoide das galhas (Meloidogyne incognita): raça 2. Revista Verde de Agroecologia e Desenvolvimento Sustentável 12:701–711.
  14. Staden, R., Beal, K. F. and Bonfield, J. K. 2000. The staden package, 1998. Bioinformatics Methods and Protocols 132:115–130.
  15. Stamatakis, A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30:1312–1313.
  16. Stimart, D. P. 1987. Genetic and physiological studies of Zinnia elegans, Z. angustifolia and their interspecific hybrids. HortScience 22:689–691.
  17. Taylor, A. L. and Netscher, C. 1974. An improved technique for preparing perineal patterns of Meloidogyne spp. Nematologica 20:268–269.
  18. Torres, A. M. 1963. Taxonomy of Zinnia New York, NY: Brittonia.
  19. Vrain, T. C., Wakarchuk, D. A., Levesque, A. C. and Hamilton, R. I. 1992. Intraspecific rDNA restriction fragment length polymorphism in the Xiphinema americanum group. Fundamental & Applied Nematology 15:563–573.
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FIGURES & TABLES

Figure 1:

(A) Perineal pattern of Meloidogyne javanica, (B) Esterase phenotypes of M. javanica detected in Zinnia elegans (J3: M. javanica; J3*: M. javanica reference isolate).

Full Size   |   Slide (.pptx)

Figure 2:

Phylogenetic relationship of Meloidogyne javanica from Ceará, Brazil, based on alignment of the sequence of the (A) D2/D3 expansion fragments of 28S rDNA and (B) sequence of ITS. The phylogenetic tree was estimated by maximum likelihood. Pratylenchus scribneri and P. zeae was used an outgroup.

Full Size   |   Slide (.pptx)

REFERENCES

  1. Ahmad, I., Whipker, B. E. and Dole, J. M. 2015. Paclobutrazol or ancymidol effects on postharvest performance of potted ornamental plants and plugs. HortScience 50:1370–1374.
  2. Ali, H. H., Fatah, K. M. and Ahmed, A. I. 2014. First record of nematode root knot on Zennia elegans caused by Meloidogyne javanica plant in Erbil Governorate, Kurdistan Region, Iraq. Arab Journal of Plant Protection 32:276–277.
  3. Carneiro, R. M. D. G. and Almeida, M. R. A. 2001. Electrophoresis technique used in the study of root-knot nematode enzymes for species identification. Nematologia Brasileira 25:555–560.
  4. Coolen, W. A. and D’Herde, C. J. 1972. A method for the quantitative extraction of nematodes from plant tissue State Agricultural and Entomology Research Station, Ghent.
  5. De Ley, P., Felix, M. A., Frisse, L. M., Nadler, S. A., Sternberg, P. W. and Thomas, W. K. 1999. Molecular and morphological characterization of two reproductively isolated species with mirror-image anatomy (Nematoda: Cephalobidae). Nematology 1:591–612.
  6. Freire, F. D. C. O. and Mosca, J. L. 2009. Patógenos associados a doenças de plantas ornamentais no Estado do Ceará. Ornamental Horticulture 15:83–89.
  7. Goeldi, E. A. 1887. Relatório sobre a moléstia do cafeeiro na provincial da Rio de Janeiro. Archivos do Museu Nacional do Rio de Janeiro 8:7–123.
  8. Jones, J. T., Haegeman, A., Danchin, E. G. J., Gaur, H. S., Helder, J., Jones, M. G. K., Kikuchi, T., Manzanilla-López, R., Palomares-Rius, J. E., Wesemael, W. M. L. and Perry, R. N. 2013. Top 10 plant-parasitic nematodes in molecular plant pathology. Molecular Plant Pathology 14:946–961.
    [PUBMED]
  9. Katoh, K. and Toh, H. 2008. Recent developments in the MAFFT multiple sequence alignment program. Briefings in Bioinformatics 9:286–298.
    [PUBMED]
  10. Katoh, S. 2013. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution 30:772–780.
    [PUBMED]
  11. Mai, W. F. and Mullin, P. G. 1996. Plant parasitic nematodes: a pictorial key to genera, New York, NY: Cornell University Press.
  12. Moens, M., Perry, R. N. and Starr, J. L. 2009. “ Meloidogyne species – a diverse group of novel and important plant parasites”, In Perry, R. N., Moens, M. and Starr, J. L. (Eds), Root-knot nematodes, Wallingford: CABI, 1–18.
  13. Moreira, F. J. C., Santos, C. D. G., Silva, G. S. and Innecco, R. 2017. Hospedabilidade de plantas ornamentais e medicinais ao nematoide das galhas (Meloidogyne incognita): raça 2. Revista Verde de Agroecologia e Desenvolvimento Sustentável 12:701–711.
  14. Staden, R., Beal, K. F. and Bonfield, J. K. 2000. The staden package, 1998. Bioinformatics Methods and Protocols 132:115–130.
  15. Stamatakis, A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30:1312–1313.
  16. Stimart, D. P. 1987. Genetic and physiological studies of Zinnia elegans, Z. angustifolia and their interspecific hybrids. HortScience 22:689–691.
  17. Taylor, A. L. and Netscher, C. 1974. An improved technique for preparing perineal patterns of Meloidogyne spp. Nematologica 20:268–269.
  18. Torres, A. M. 1963. Taxonomy of Zinnia New York, NY: Brittonia.
  19. Vrain, T. C., Wakarchuk, D. A., Levesque, A. C. and Hamilton, R. I. 1992. Intraspecific rDNA restriction fragment length polymorphism in the Xiphinema americanum group. Fundamental & Applied Nematology 15:563–573.

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