Table 1.
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 |
Table 2.
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″ N 076°09′36.9″ W | 2,963 | 13.9 | 0.0 | 0.0 | 0.0 |
| Pasture bordering annual crops (PBAC) | 02°56′57,3″ N 076°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″ N 076°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″ N 076°12′41,3″ W | 1,894 | 18.0 | 0.0 | 0.0 | 0.0 |
| Strawberry cultivation (SC) | 02°58′43.2″ N 076°09′36.9″ W | 2,963 | 13.9 | 0.0 | 0.0 | 0.0 |
| Combined lulo and papaya cultivation (CLPC) | 02°56′0,39″ N 076°17′15″ W | 1,820 | 18.5 | 0.0 | 0.0 | 0.0 |
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).
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.
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.
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).