<abstract xmlns="http://www.w3.org/1999/xhtml"><p>Samples of black pepper root with the presence of galls from the most southern region of the State of Bahia, Brazil, were characterized biochemically and morphologically using three criteria: i) observation of the anterior region of the males; ii) analysis of female perineal configuration and iii) polyacrylamide gel electrophoresis technique. <italic>Meloidogyne arenaria</italic> was found. This is the first report of this pathosystem in the State.</p></abstract>

Samples of black pepper root with the presence of galls from the most southern region of the State of Bahia, Brazil, were characterized biochemically and morphologically using three criteria: i) observation of the anterior region of the males; ii) analysis of female perineal configuration and iii) polyacrylamide gel electrophoresis technique. Meloidogyne arenaria was found. This is the first report of this pathosystem in the State.


<div xmlns="http://www.w3.org/1999/xhtml">
<sec id="j_helm-2021-0015_s_001"><div>Introduction</div>
<p>Black pepper (<italic>Piper nigrum</italic> L.), originally from India, is one of the most consumed spices worldwide, being widely used as a condiment in food preparation and processing (<a ref-type="bibr" href="#j_helm-2021-0015_ref_018">Nair, 2011</a>). It is currently grown on a commercial scale in 26 countries, mainly Indonesia, Malaysia, Brazil, Vietnam, China and Thailand (Ravindran, 2000). Brazil stands out as the world’s third largest producer of black pepper, with cultivation growing 53 % in 2017, as the country has ideal conditions for its development, especially in the states of Espirito Santo, Southern Bahia and Pará, the latter being the largest national producer and responsible for almost 70 % of Brazilian production (<a ref-type="bibr" href="#j_helm-2021-0015_ref_011">IBGE, 2017</a>).</p>
<p>Despite the growing production, pipericulture faces serious phytosanitary problems, mainly due to the occurrence of soil pathogens such as fungi and root-knot nematodes, <italic>Meloidogyne</italic> spp., Goeldi, pathogens that are diffi cult to manage due to their wide dissemination and host range (<a ref-type="bibr" href="#j_helm-2021-0015_ref_001">Antes <italic>et al</italic>., 2012</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_015">Maleita <italic>et al</italic>., 2012</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_019">Negretti <italic>et al</italic>., 2014</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_002">Bellé <italic>et al</italic>., 2017</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_017">Moreira <italic>et al</italic>., 2017</a>). In addition, phytonematodes are considered the main elements of predisposition to fusariosis, also known as root rot, a disease caused by the fungus <italic>Fusarium solani</italic> f. sp. <italic>piperis</italic> anti, 2015).</p>
<p>Considering the importance that root-knot nematodes represent and the lack of consistent information about the distribution and occurrence of these pathogens in black pepper, this work aimed to characterize, biochemically and morphologically, the gall nematode species associated with black pepper plantations in the most southern region of the State of Bahia, Brazil.</p>
</sec>
<sec id="j_helm-2021-0015_s_002"><div>Material and Methods</div>
<p>The study was carried out at Fazenda Conjunto Bom Jardim, located in the municipality of Itabela, in the extreme south of Bahia, Brazil, on a property composed of 10 ha grown with black pepper (latitude 16 ° 44 ‘45’ ‘S and longitude 39 ° 28’ 58 ‘’ W), altitude of 120 m at sea level, with four years of age and 3.70 x 1.65 m spacing. Three areas were selected, each consisting of 1 ha, two of them with cv. Kottanadan Roxa and the other with cv. Bragantina. The samples were collected from 0 to 30 cm deep, in the rhizosphere region, giving preference to the collection of radicellae from weakened plants (<a ref-type="fig" href="#j_helm-2021-0015_fig_001">Fig. 1</a>).</p>
<figure id="j_helm-2021-0015_fig_001" position="float" fig-type="figure">
<h2>Fig. 1</h2>
<figCaption><p>(A) Weakened black pepper plants grown in the municipality of Itabela in the extreme south of Bahia; (B) Roots with gall symptoms induced by gall nematodes. Photo: Rafael Assis de Souza.</p></figCaption>
<img xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="graphic/j_helm-2021-0015_fig_001.jpg" src="https://sciendo-parsed.s3.eu-central-1.amazonaws.com/6471e8c9215d2f6c89db4b8b/j_helm-2021-0015_fig_001.jpg?X-Amz-Algorithm=AWS4-HMAC-SHA256&amp;X-Amz-Date=20240704T222835Z&amp;X-Amz-SignedHeaders=host&amp;X-Amz-Expires=18000&amp;X-Amz-Credential=AKIA6AP2G7AKP25APDM2%2F20240704%2Feu-central-1%2Fs3%2Faws4_request&amp;X-Amz-Signature=53f2eadb795b6013773d921e11490ad1a82fbc44bb70f1dd495251434d7b4be3" class="mw-100"/>
</figure>
<p>After zig-zag walking in each area, 10 subsamples were collected that made up 1 composite sample for each area.</p>
<p>The samples were packed in plastic bags, duly identified, and sent to the Laboratory of Nematology at the Research Center for Plant Protection and Animal Health, of the Instituto Biológico, in Campinas, SP, for extraction and identification of phytoparasitic nematodes.</p>
<p>The nematodes were extracted from 10g of roots using the method of <a ref-type="bibr" href="#j_helm-2021-0015_ref_006">Coolen and D´Herde (1972)</a> and the quantification was performed in Peters slide.</p>
<p>The identification of the <italic>Meloidogyne</italic> species was carried out considering three criteria: i) observation of the anterior region of the males; ii) analysis of female perineal configuration (<a ref-type="bibr" href="#j_helm-2021-0015_ref_010">Hunt &amp; Handoo, 2009</a>) and iii) polyacrylamide gel electrophoresis technique according to <a ref-type="bibr" href="#j_helm-2021-0015_ref_020">Oliveira <italic>et al</italic>. (2012)</a>.</p>
<p>Thirty-five females were used in the isoenzymatic identification (esterase) considering five females per sample, totaling seven samples. Five females were also used for identification by the perineal configuration and five males for observation of the anterior region of the nematode.</p>
<sec id="j_helm-2021-0015_s_002_s_001"><div>Ethical Approval and/or Informed Consent</div>
<p>For this study formal consent is not required.</p>
</sec>
<sec id="j_helm-2021-0015_s_002_s_002"><div>Results and Discussion</div>
<p>Black pepper plants parasitized by gall nematodes generally become chlorotic, with severe leaf yellowing, defoliation and reduction in chlorophyll content, which affects vegetative development, resulting in significant reduction in yield and even death (<a ref-type="bibr" href="#j_helm-2021-0015_ref_009">Freire &amp; Monteiro, 1978</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_008">Ferraz &amp; Lordello, 1989</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_016">Mohandas &amp; Ramana, 1991</a>). Some of these symptoms, such as yellowing, defoliation and roots with numerous galls were observed in the plants analyzed in our study.</p>
<p>Studying the anterior region of the males and analyzing the perineal configuration of the females, the presence of <italic>Meloidogyne arenaria</italic> was confirmed (<a ref-type="fig" href="#j_helm-2021-0015_fig_002">Figs. 2A and 2B)</a>.</p>
<figure id="j_helm-2021-0015_fig_002" position="float" fig-type="figure">
<h2>Fig. 2</h2>
<figCaption><p>(A) Anterior region of the males and (B) Perineal configuration of the characteristic females of <italic>Meloidogyne arenaria</italic>.</p></figCaption>
<img xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="graphic/j_helm-2021-0015_fig_002.jpg" src="https://sciendo-parsed.s3.eu-central-1.amazonaws.com/6471e8c9215d2f6c89db4b8b/j_helm-2021-0015_fig_002.jpg?X-Amz-Algorithm=AWS4-HMAC-SHA256&amp;X-Amz-Date=20240704T222835Z&amp;X-Amz-SignedHeaders=host&amp;X-Amz-Expires=18000&amp;X-Amz-Credential=AKIA6AP2G7AKP25APDM2%2F20240704%2Feu-central-1%2Fs3%2Faws4_request&amp;X-Amz-Signature=5331dcd194fe345dba4aaf66411cb02339f3d0dac183840e517c218ac9458474" class="mw-100"/>
</figure>
<p>We observed females with perineal pattern rounded to ovoid, with thin to thick streaks. Low dorsal arch, flattened, with smooth or slightly wavy streaks, slightly bent towards the tail tip on the lateral line; usually forming shoulders on the lateral portion of the arch. The dorsal and ventral striations are usually at an angle on the lateral lines; distinct lateral field, slightly irregular. In males, the labial region was continuous with the rest of the body, smooth, labial disc more or less rounded, slightly elevated above the level of the medial lips. Long distance from the orifice of the dorsal esophageal gland to the base of the stylet.</p>
<p>The analysis of polyacrylamide gel electrophoresis also revealed a characteristic profile of <italic>M. arenaria</italic> (<a ref-type="fig" href="#j_helm-2021-0015_fig_003">Fig. 3</a>).</p>
<figure id="j_helm-2021-0015_fig_003" position="float" fig-type="figure">
<h2>Fig. 3</h2>
<figCaption><p>Phenotype of polyacrylamide gel esterase, characteristic of <italic>Meloidogyne arenaria</italic>, obtained from females of the nematode of galls extracted from black pepper roots. Mj = <italic>M. javanica</italic> (standard); 2 to 7 = <italic>M. arenaria</italic></p></figCaption>
<img xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="graphic/j_helm-2021-0015_fig_003.jpg" src="https://sciendo-parsed.s3.eu-central-1.amazonaws.com/6471e8c9215d2f6c89db4b8b/j_helm-2021-0015_fig_003.jpg?X-Amz-Algorithm=AWS4-HMAC-SHA256&amp;X-Amz-Date=20240704T222835Z&amp;X-Amz-SignedHeaders=host&amp;X-Amz-Expires=18000&amp;X-Amz-Credential=AKIA6AP2G7AKP25APDM2%2F20240704%2Feu-central-1%2Fs3%2Faws4_request&amp;X-Amz-Signature=08997f8d467aac5cc0a1938ae53da68019add6bd90a10cdfabf339dbd681ab6a" class="mw-100"/>
</figure>
<p>The numbers of second stage juveniles (J2) + eggs extracted from black pepper roots are shown in <a ref-type="table" href="#j_helm-2021-0015_tab_001">Table 1</a>. Both cultivars showed a high number of <italic>M. arenaria</italic> demonstrating the susceptibility of these cultivars to this species.</p>
<table-wrap id="j_helm-2021-0015_tab_001" position="float">
<label>Table 1</label>
<caption><p>Number of second stage juveniles (J2) and eggs of <italic>Meloidogyne arenaria</italic> obtained in 10 g of root from black pepper cv. Kottanadan roxa and Bragantina in cultivation areas in the extreme south of the state of Bahia, BA.</p></caption>
<table frame="hsides" rules="groups">
<thead><tr><th align="center"></th>
<th align="center">Area 1<break></break>‘Kottanadan Roxa’</th>
<th align="center">Area 2<break></break>‘Kottanadan Roxa’</th>
<th align="center">Area 3<break></break>‘Bragantina’</th>
</tr></thead>
<tbody>
<tr><td align="center"></td>
<td align="center">root</td>
<td align="center">root</td>
<td align="center">root</td>
</tr>
<tr><td align="center">J2</td>
<td align="center">5600</td>
<td align="center">5300</td>
<td align="center">5000</td>
</tr>
<tr><td align="center">eggs</td>
<td align="center">5600</td>
<td align="center">4300</td>
<td align="center">3100</td>
</tr>
</tbody>
</table>
</table-wrap>
<p>The first study that showed the presence of <italic>Meloidogyne</italic> spp. in black pepper plantations was carried out in 1901 by Zimmerman cited by <a ref-type="bibr" href="#j_helm-2021-0015_ref_022">Winoto (1972)</a>.</p>
<p>In Brazil, reports involving the black pepper pathosystem x <italic>Meloidogyne</italic> spp. are rare and mostly related to <italic>M. incognita</italic> in the State of Pará (<a ref-type="bibr" href="#j_helm-2021-0015_ref_014">Lordello &amp; Silva, 1974</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_013">Ichinohe, 1976</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_008">Freire &amp; Monteiro 1978</a>).</p>
<p>In Brazil, the only report of <italic>M. arenaria</italic> in black pepper is from 1965, made by Dr. Jair C. Carvalho, based on the analysis of soil and roots from Caraguatatuba, SP (<a ref-type="bibr" href="#j_helm-2021-0015_ref_004">Carvalho, 1965</a>). It is important to highlight that before the development of the isoenzyme electrophoresis technique, the identification of <italic>Meloidogyne</italic> species was performed exclusively by examining the perineal configuration (<a ref-type="bibr" href="#j_helm-2021-0015_ref_005">Chitwood, 1949</a>). It is known, however, that this technique is extremely subjective, and it is of fundamental importance also to characterize these pathogens by biochemical means, through the technique of isoenzyme electrophoresis (<a ref-type="bibr" href="#j_helm-2021-0015_ref_003">Carneiro &amp; Almeida, 2001</a>; <a ref-type="bibr" href="#j_helm-2021-0015_ref_020">Oliveira <italic>et al</italic>. 2012</a>), and by observation of the region of the males, as performed in the present study. The use of these last two techniques associated with the analysis of the perineal configuration brings certainty regarding the correct diagnosis of <italic>Meloidogyne</italic> spp. (<a ref-type="bibr" href="#j_helm-2021-0015_ref_007">Cunha <italic>et al</italic>, 2018</a>).</p>
<p>Considering that the success of any nematode management strategy depends, initially, on the knowledge of the species present in the areas where it is desired to reduce the population of these pathogens, the information obtained in the present work may be useful in the development and adoption of appropriate techniques for the management of parasitic nematodes of black pepper.</p>
<p>The species of gall nematode associated with black pepper cultivation in the extreme south of the State of Bahia is <italic>Meloidogyne arenaria</italic>. It is the first report of this pathosystem in the State, and therefore expands its geographical scope.</p>
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</div>

Occurrence of <italic>Meloidogyne arenaria</italic> in black pepper (<italic>Piper nigrum</italic> L.) in the extreme south of the State of Bahia, Brazil

Helminthologia

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Occurrence of  in black pepper ( L.) in the extreme south of the State of Bahia, Brazil

Samples of black pepper root with the presence of galls from the most southern region of the State of Bahia, Brazil, were characterized biochemically and...

Occurrence of Meloidogyne arenaria in black pepper (Piper nigrum L.) in the extreme south of the State of Bahia, Brazil

Article Category: Research Note

Published Online: Jun 08, 2021

Page range: 213 - 216

Received: May 20, 2020

Accepted: Dec 04, 2020

DOI: https://doi.org/10.2478/helm-2021-0015

Keywords
nematological survey, pipericulture, root-knot nematode

© 2021 R. A. De Souza, F. R. Alves, C. M. G. De Oliveira, J. M. O. Rosa, J. De Assis Pinheiro, G. P. De Aguiar, W. B. Moraes, published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Occurrence of Meloidogyne arenaria in black pepper (Piper nigrum L.) in the extreme south of the State of Bahia, Brazil

Article Category: Research Note

Published Online: Jun 08, 2021

Page range: 213 - 216

Received: May 20, 2020

Accepted: Dec 04, 2020

DOI: https://doi.org/10.2478/helm-2021-0015

Keywordsnematological survey, pipericulture, root-knot nematode

© 2021 R. A. De Souza, F. R. Alves, C. M. G. De Oliveira, J. M. O. Rosa, J. De Assis Pinheiro, G. P. De Aguiar, W. B. Moraes, published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Keywords
nematological survey, pipericulture, root-knot nematode