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Isolation, identification, and pathogenicity of Steinernema carpocapsae and its bacterial symbiont in Cauca-Colombia

Esteban Neira-Monsalve
Esteban Neira-Monsalve
, 
Natalia Carolina Wilches-Ramírez
Natalia Carolina Wilches-Ramírez
, 
Wilson Terán
Wilson Terán
, 
María del Pilar Márquez
María del Pilar Márquez
, 
Ana Teresa Mosquera-Espinosa
Ana Teresa Mosquera-Espinosa
 und 
Adriana Sáenz-Aponte
Adriana Sáenz-Aponte
   | 31. Aug. 2020
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Online veröffentlicht: 31. Aug. 2020
Seitenbereich: 1 - 16
DOI: https://doi.org/10.21307/jofnem-2020-089
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© 2020 Esteban Neira-Monsalve et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Phylogenetic tree of S. carpocapsae BPS. A. Phylogenetic relationships from the 18S marker. B. Phylogenetic relationships from concatenated ITS and 28S markers. The relationships were inferred by maximum likelihood estimation for the 18S marker while UPGMA was used for the concatenated markers. Caenorhabditis. elegans was used as an outgroup. The numbers on the nodes indicate the bootstrap values (1,000 replications).
Phylogenetic tree of S. carpocapsae BPS. A. Phylogenetic relationships from the 18S marker. B. Phylogenetic relationships from concatenated ITS and 28S markers. The relationships were inferred by maximum likelihood estimation for the 18S marker while UPGMA was used for the concatenated markers. Caenorhabditis. elegans was used as an outgroup. The numbers on the nodes indicate the bootstrap values (1,000 replications).

Figure 2:

Susceptibility of two Hass avocado (P. americana) pest insects to S. carpocapsae BPS and S. carpocapsae FA2015. A. E. cucumeris. B. P. cinereus.
Susceptibility of two Hass avocado (P. americana) pest insects to S. carpocapsae BPS and S. carpocapsae FA2015. A. E. cucumeris. B. P. cinereus.

Figure 3:

Microscopic and macroscopic morphological characteristics of the bacterial symbiont, Xenorhabdus nematophila BPS. A. Gram stain, 100X. B. Colonies on NBTA agar. C. Colony on MacConkey agar. D. Isolate of bacterial symbiont on blood agar, view from the back of the box. The bar measures 0.5 cm.
Microscopic and macroscopic morphological characteristics of the bacterial symbiont, Xenorhabdus nematophila BPS. A. Gram stain, 100X. B. Colonies on NBTA agar. C. Colony on MacConkey agar. D. Isolate of bacterial symbiont on blood agar, view from the back of the box. The bar measures 0.5 cm.

Figure 4:

Phylogenetic tree of the bacterial symbiont Xenorhabdus nematophila BPS. A. Phylogenetic relationships from the 16S marker. B. Phylogenetic relationships from concatenation of dnaN, recA, gltX, and rplB markers. The relationships were inferred by maximum likelihood estimation. Morganella morganii subsp. morganii was used as an outgroup for 16S, while Proteus mirabilis and Photorhabdus luminescens subsp. laumondii were used for concatenated markers. The numbers on the nodes indicate the bootstrap values (≥ 50%) (100 replicates).
Phylogenetic tree of the bacterial symbiont Xenorhabdus nematophila BPS. A. Phylogenetic relationships from the 16S marker. B. Phylogenetic relationships from concatenation of dnaN, recA, gltX, and rplB markers. The relationships were inferred by maximum likelihood estimation. Morganella morganii subsp. morganii was used as an outgroup for 16S, while Proteus mirabilis and Photorhabdus luminescens subsp. laumondii were used for concatenated markers. The numbers on the nodes indicate the bootstrap values (≥ 50%) (100 replicates).

Accession numbers of sequences used for phylogenetic analysis of the EPN isolated and its bacterial symbiont.

Gen
Species 18S ITS 28S
Steinernema carpocapsae BPS MK558002 ■ MK558041 ■ MK558056 ■
Steinernema carpocapsae KJ636405 AF121049 KJ950293
Steinernema arenarium KJ636393 KU194614 KU194619
Steinernema feltiae KJ636413 AF121050 JF920963
Steinernema glaseri KU180674 AF122015 GU177831
Steinernema bicornutum KT878311 AF121048 GU569045
Steinernema poinari KT878314 KF241753 KF241750
Steinernema akhursti KT878310 DQ375757 KF289902
Steinernema karii AJ417021 AY230173 AF331902
Steinernema beitlechemi KT878316 KT373856 KT580949
Steinernema kushidai LC157426 GQ497741 AF331897
Steinernema affine FJ040425 AY230159 AF331899
Steinernema websteri – FJ381666 AY841762
Steinernema scarabaei – FJ263673 AY172023
Steinernema khuongi – GU174002 GU177835
Steinernema diaprepesi – AF122021 GU177828
Steinernema hermaphroditum – MF663703 MF693228
Steinernema rarum – DQ221115 AY253296
Steinernema intermedium – AF122016 AF331909
Steinernema unicornum – GQ497167 GU191462
Steinernema scapterisci – AF122020 AF331898
Steinernema neocurtillae – AF122018 FJ263674
Steinernema monticolum – AF122017 AF331895
Steinernema nguyeni – KP325084 KR815816
Steinernema sacchari – KC633095 KC633096
Steinernema phyllophagae – FJ410327 FJ666054
Steinernema pakistanense – MF289981 JX068823
Steinernema australe – FJ235125 FJ235126
Steinernema brazilense – FJ410325 FJ410326
Steinernema riobrave – AY230182 GU177834
Steinernema cubanum – AY230166 AF331889
Steinernema ceratophorum – AY230165 AF331888
Steinernema texanum – EF152568 EF152569
Steinernema cholashanense – MF039642 EF520284
Steinernema apuliae – HQ416968 KU194621
Steinernema khoisanae – DQ314287 DQ314289
Steinernema aciari – AY787660 GU395637
Steinernema citrae – FJ235074 MF540678
Steinernema silvaticum – MG543848 MG547579
Steinernema surkhetense – MF919614 KU187262
Steinernema litorale – JF892546 JQ795723
Steinernema anatoliense – EU200356 AY841761
Caenorhabditis elegans X03680 X03680 X03680
Panagrellus redivivus – – AF331910
16S dnaN recA gltX rplB
Xenorhabdus nematophila BPS MK558196 ■ MK570079 ■ MK570081 ■ MK570080 ■ MK570082 ■
Xenorhabdus nematophila ATCC19061 D78009 NC_014228 AF127333 NC_014228 CBJ88402
Xenorhabdus beddingii AY278675 FJ831460 FJ823415 FJ840506 FJ808999
Xenorhabdus bovienii X82252 FJ831466 FJ823426 FJ840514 FJ809005
Xenorhabdus japonica DQ202310 FJ831453 FJ823400 FJ840503 FJ808989
Xenorhabdus poinarii D78010 FJ831454 FJ823409 FJ840499 FJ808995
Xenorhabdus szentirmaii AJ810295 FJ831458 FJ823416 FJ840508 FJ809002
Xenorhabdus ehlersii AJ810294 FJ831448 FJ823398 FJ840495 FJ808992
Xenorhabdus innexi AJ810292 FJ831476 FJ823424 FJ840522 FJ809018
Xenorhabdus budapestensis AJ810293 FJ831474 FJ823418 FJ840518 FJ809017
Xenorhabdus indica AM040494 FJ831470 FJ823421 FJ840520 FJ809013
Xenorhabdus cabanillasii AY521244 FJ831472 FJ823422 FJ840521 FJ809015
Xenorhabdus doucetiae DQ211709 FJ831450 FJ823402 FJ840497 FJ809001
Xenorhabdus griffiniae DQ211710 FJ831449 FJ823399 FJ840496 FJ808991
Xenorhabdus hominickii DQ211719 FJ831461 FJ823410 FJ840510 FJ809010
Xenorhabdus koppenhoeferi DQ205450 FJ831457 FJ823413 FJ840504 FJ809004
Xenorhabdus kozodoii DQ211716 FJ831446 FJ823404 FJ840493 FJ808994
Xenorhabdus mauleonii DQ211715 FJ831464 FJ823417 FJ840507 FJ809003
Xenorhabdus miraniensis DQ211713 FJ831459 FJ823414 FJ840505 FJ808990
Xenorhabdus romanii DQ211717 FJ831451 FJ823403 FJ840498 FJ809000
Xenorhabdus stockiae DQ202309 FJ831477 FJ823425 FJ840524 FJ809020
Xenorhabdus vietnamensis DQ205447 FJ831452 FJ823401 FJ840502 FJ808998
Xenorhabdus ishibashii GQ149086 JQ348908 JQ348906 JQ348909 PHM62307
Xenorhabdus magdalenensis HQ877464 JF798399 JF798401 JF798400 -
Xenorhabdus khoisanae HQ142625 AB685733 AB685736 AB685734 KMJ46808
Xenorhabdus thuongxuanensis KX602193 KX602195 KX602194 KX602196 OKP02162
Xenorhabdus eapokensis KX602187 KX602189 KX602188 KX602190 OKP00696
Morganella morganii subsp. morganii AJ301681 – – – –
Photorhabdus luminescens subsp. laumondii – FJ831497 KT963835 KT963845 FJ817457
Proteus mirabilis – NC_010554 X14870 CAR43779 CAR46384

Isolation of EPNs according to types of vegetation cover at sampling sites.

Vegetation cover Location Altitude (m.a.s.l) Soil temperature (°C) Recovery frequency (%) Larvae of G. mellonella affected by EPNs (%) Larvae of T. molitor affected by EPNs (%)
Natural grassland (NG) 02°58′43.2″ N076°09′36.9″ W 2,963 13.9 0.0 0.0 0.0
Pasture bordering annual crops (PBAC) 02°56′57,3″ N076°16′21,5″ W 1,757 20.0 0.0 0.0 0.0
Natural forest (NF) 02°56′0.99″ N 76°17′19.8″ W 1,863 16.4 50.0 21.7 1.7
First-growth bamboo (FGAB) 03°00′49.5″ N076°12′41,3″ W 1,894 18.0 0.0 0.0 0.0
Coffee cultivation (CC) 02°56′0.45″ N 76°17′19.1″ W 1,852 16.6 8.3 3.3 1.7
Horticultural cultivation (HC) 03°00′49.5″ N076°12′41,3″ W 1,894 18.0 0.0 0.0 0.0
Strawberry cultivation (SC) 02°58′43.2″ N076°09′36.9″ W 2,963 13.9 0.0 0.0 0.0
Combined lulo and papaya cultivation (CLPC) 02°56′0,39″ N076°17′15″ W 1,820 18.5 0.0 0.0 0.0
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