First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus
Нематоды Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 были выделены, с использованием гусениц Galleria mellonella в качестве приманок, из образцов почвы, собранных около Краснодара (Северный Кавказ). Обнаружены различия между краснодарской и типовой (югославской) культурами S. bicornutu...
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Інститут зоології ім. І. І. Шмальгаузена НАН України
2003
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| Cite this: | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Caucasus / E. S. Ivanova, S. E. Spiridonov // Вестн. зоологии. — 2003. — Т. 37, № 3. — С. 57-63. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859980118064103424 |
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| author | Ivanova, E.S. Spiridonov, S.E. |
| author_facet | Ivanova, E.S. Spiridonov, S.E. |
| citation_txt | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Caucasus / E. S. Ivanova, S. E. Spiridonov // Вестн. зоологии. — 2003. — Т. 37, № 3. — С. 57-63. — англ. |
| collection | DSpace DC |
| description | Нематоды Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 были выделены, с использованием гусениц Galleria mellonella в качестве приманок, из образцов почвы, собранных около Краснодара (Северный Кавказ). Обнаружены различия между краснодарской и типовой (югославской) культурами S. bicornutum в средней длине инвазионных личинок и длине спикул самцов. Все эти различия (за исключением длины спикул у самцов второй генерации) не являются статистически достоверными. Рестрикционный анализ рибосомальной ДНК дифференцирует эти две популяции S. bicornutum лишь по спектрам эндонуклеазы Cfo I (один сайт рестрикции у типовой, и два — у краснодарской культуры). Проведено секвенирование 715 пар оснований фрагмента рибосомального гена (ITS1 + 5.8S + ITS2). Сравнение с последовательностью типового изолята показало различие в 18 парах нуклеотидов, включая 17 замен и одну делецию в типовой культуре. Выявленные морфологические и молекулярные различия оцениваются как межпопуляционные, а обнаруженные в Краснодаре нематоды как еще один изолят S. bicornutum.
Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 was isolated by Galleria baiting from soil samples collected near Krasnodar (North Caucasus). The differences were found between the type (Yugoslavian) population and Krasnodar isolate in the average length of infective juveniles and spicules. All these differences (except spicule length in the second generation males) are not statistically significant. The RFLP analysis was able to distinguish nematodes from these two populations of S. bicornutum only in the Cfo I profiles (one restriction site in the type culture and two restriction sites in the Krasnodar isolate). The 715 bp long fragment of ribosomal gene (ITS1 + 5.8S + ITS2) of the Krasnodar isolate of S. bicornutum was sequenced and compared with similar sequence of the type strain. In accordance with RFLP data, one additional Cfo I restriction site was found in the Krasnodar S. bicornutum. Totally, the difference in 18 pairs of nucleotides per fragment of 715 bp was found between Krasnodar and type cultures, including 17 substitutions and one deletion (in type culture). Reported morphological and molecular differences are considered as interpopulational ones, and described nematodes from Krasnodar as another isolate of S. bicornutum.
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ÓÄÊ 576.895.132
FIRST REPORT OF STEINERNEMA BICORNUTUM
(NEMATODA, RHABDITIDA, STEINERNEMATIDAE)
FROM THE NORTH CAUCASUS
E. S. Ivanova, S. E. Spiridonov
Institute of Parasitology, Russian Academy of Sciences, Leninskii pr., 33, Moscow, 117071 Russia
E-mail: rjnem@rjnem.msk.ru
Accepted 17 February 2002
Ïåðâàÿ íàõîäêà Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) íà Ñåâåðíîì
Êàâêàçå. Èâàíîâà Å. Ñ., Ñïèðèäîíîâ Ñ. Ý. — Íåìàòîäû Steinernema bicornutum Tallosi, Peters &
Ehlers, 1995 áûëè âûäåëåíû, ñ èñïîëüçîâàíèåì ãóñåíèö Galleria mellonella â êà÷åñòâå ïðèìàíîê,
èç îáðàçöîâ ïî÷âû, ñîáðàííûõ îêîëî Êðàñíîäàðà (Ñåâåðíûé Êàâêàç). Îáíàðóæåíû ðàçëè÷èÿ
ìåæäó êðàñíîäàðñêîé è òèïîâîé (þãîñëàâñêîé) êóëüòóðàìè S. bicornutum â ñðåäíåé äëèíå èí-
âàçèîííûõ ëè÷èíîê è äëèíå ñïèêóë ñàìöîâ. Âñå ýòè ðàçëè÷èÿ (çà èñêëþ÷åíèåì äëèíû ñïèêóë
ó ñàìöîâ âòîðîé ãåíåðàöèè) íå ÿâëÿþòñÿ ñòàòèñòè÷åñêè äîñòîâåðíûìè. Ðåñòðèêöèîííûé àíàëèç
ðèáîñîìàëüíîé ÄÍÊ äèôôåðåíöèðóåò ýòè äâå ïîïóëÿöèè S. bicornutum ëèøü ïî ñïåêòðàì ýí-
äîíóêëåàçû Cfo I (îäèí ñàéò ðåñòðèêöèè ó òèïîâîé, è äâà — ó êðàñíîäàðñêîé êóëüòóðû).
Ïðîâåäåíî ñåêâåíèðîâàíèå 715 ïàð îñíîâàíèé ôðàãìåíòà ðèáîñîìàëüíîãî ãåíà (ITS1 + 5.8S +
ITS2). Ñðàâíåíèå ñ ïîñëåäîâàòåëüíîñòüþ òèïîâîãî èçîëÿòà ïîêàçàëî ðàçëè÷èå â 18 ïàðàõ íóê-
ëåîòèäîâ, âêëþ÷àÿ 17 çàìåí è îäíó äåëåöèþ â òèïîâîé êóëüòóðå. Âûÿâëåííûå ìîðôîëîãè÷åñêèå
è ìîëåêóëÿðíûå ðàçëè÷èÿ îöåíèâàþòñÿ êàê ìåæïîïóëÿöèîííûå, à îáíàðóæåííûå â Êðàñíîäàðå
íåìàòîäû êàê åùå îäèí èçîëÿò S. bicornutum.
Êëþ÷åâûå ñ ëîâ à: ýíòîìîïàòîãåííûå íåìàòîäû, Rhabditida, Steinernema, S. bicornutum, íî-
âûé èçîëÿò, ìîðôîëîãèÿ, rDNA.
First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-
casus. Ivanova E. S., Spiridonov S. E. — Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 was
isolated by Galleria baiting from soil samples collected near Krasnodar (North Caucasus). The diffe-
rences were found between the type (Yugoslavian) population and Krasnodar isolate in the average
length of infective juveniles and spicules. All these differences (except spicule length in the second
generation males) are not statistically significant. The RFLP analysis was able to distinguish nematodes
from these two populations of S. bicornutum only in the Cfo I profiles (one restriction site in the type
culture and two restriction sites in the Krasnodar isolate). The 715 bp long fragment of ribosomal gene
(ITS1 + 5.8S + ITS2) of the Krasnodar isolate of S. bicornutum was sequenced and compared with
similar sequence of the type strain. In accordance with RFLP data, one additional Cfo I restriction site
was found in the Krasnodar S. bicornutum. Totally, the difference in 18 pairs of nucleotides per fragment
of 715 bp was found between Krasnodar and type cultures, including 17 substitutions and one deletion
(in type culture). Reported morphological and molecular differences are considered as inter-populatio-
nal ones, and described nematodes from Krasnodar as another isolate of S. bicornutum.
K e y wo r d s: entomopathogenic nematodes, Rhabditida, Steinernema, S. bicornutum, new isolate,
morphology, rDNA.
The studies in the taxonomy of Steinernematidae — the family of entomopathogenic soil-inhabiting ne-
matodes — were very active in Western Europe because of expected economic potential of these nematodes
as new tool for pest management. Unusual species of this family was found in the province of Vojvodina
(Northern Yugoslavia) and described, as Steinernema bicornutum (Tallosi, Peters & Ehlers, 1995). The pecu-
liarity of this species which was not known before among steinernematids is the presence of two horn-like
appendages on the anterior end of infective juveniles. Since then, this species was reported throughout the
Europe, though nowhere it was found common or widespread. Recently, the nematodes with morphology of
infective juveniles similar to S. bicornutum, were isolated during Galleria baiting survey of Krasnodar region
in Southern Russia (Kuban river on the north slope of Caucasus). After study and comparison these nema-
todes were considered as conspecific with Yugoslavian S. bicornutum. Short description of differences between
type and Krasnodar cultures of this species are presented below.
Vestnik zoologii, 37(3): 57–63, 2003
© E. S. Ivanova, S. E. Spiridonov, 2003
Material and methods
Steinernema bicornutum culture was isolated from the garden soil on the grounds of the All-Russian In-
stitute for Biological Protection of Plants in the vicinities of Krasnodar in June 2000. Culture was maintained
in Moscow on Galleria. Adults for morphological examination were obtained from infected caterpillars. All
measurements of nematodes given in text are in micrometers. Infective juveniles were stored in the refrige-
rator and used for molecular studies. For comparative studies the isolate of S. bicornutum was also obtained
in April 2000 from Dr. Igor Kramer (then at Federal Agricultural Centre in Wadenswil, Switzerland).
Juveniles were collected in 8 mkl of worm-lysis buffer (100 mM KCl, 20 mM Tris-HCl pH 8.3, 3 mM
MgCl2, 2 mM DDT and 0,9% Tween 20) and homogenized. The contiguous sequence including 2 non-tran-
scribed regions and 5.8S gene (ITS1 + 5.8S + ITS2) was amplified for compared Krasnodar and Swiss po-
pulations of S. bicornutum with the use of Vrain et al. (1992) primers. The RFLP analysis of obtained PCR
products was performed as described by Hominick et al. (1997) with 7 endonucleases (Dde I; Eco RI; Cfo I;
Hind III; Hinf I; Rsa I and Pvu II). To obtain product for sequencing of Krasnodar isolate a pair or primers
TW81 (5'–GTTTCAGTAGGTGAACCTGA-3') and AB28 (5'–ATATGATTAAGTTCAGCGGGT-3') as
described by Joyce et al. (1994) was used. Amplifications were carried out in a 23–25 mkl reaction volume,
containing 2.5 mkl of 10x PCR-buffer solution, 5 mkl of Q-solution, 0.5 mkl of dNTP mix, 0.15 mkl of each
primer and 0.1 mkl of Taq polymerase (from standard "Taq PCR Core Kit" of Qiagen Ltd, Crawley, UK)
with 2–4 mkl of the single juvenile homogenate and 12.6 mkl of autoclaved bidistilled water. PCR products
were cleaned using Qiaquick PCR or Gel Purification Kit (Qiagen Ltd, Crawley, UK). Attempt was made
to use purified PCR products for direct sequencing using AB28, TW81, but as direct sequencing of two
samples of S. bicornutum from Krasonodar produced electrophoregrams with multiple peaks, cloning of PCR
products was performed. PCR product from single juvenile was cloned in pGEM-T vector and transformed
into JM109 competent cells according to the instructions of the manufacturer (Promega, Benelux b. v.
Leiden, The Netherlands). PCR products from two colonies were used for sequencing. For the sequencing
reaction the mixture of 8 mkl BigDye Terminator v3.0 cycle sequencing ready reaction mix (Applied
Biosystems, Warrington, UK) and 0.5 mkl of each primer were used with 1–3 mkl of PCR product. Then
the total volume of sequencing mix was adjusted to the total volume of 20 mkl with double distilled water.
Nucleotides and other DNA remnants were removed from sequencing Gel Filtration cartridges (Edge
Biosystems, Inc., Gaithersburg, USA).
The contiguous sequences including 2 non-transcribed regions (ITS) and 5.8S gene obtained from two
colonies of transformants with Krasnodar S. bicornutum were found to be identical. The sequence was depo-
sited in the GenBank (accession number AY 171 279). Similar sequence for type (Yugoslavian) culture of
S. bicornutum was obtained from GenBank (Accession number AF 121 048). These sequences were compared
using GeneDoc 2.5.000 program.
Results
Gene r a l mo r pho l o g y. Body robust. Head end bluntly rounded or truncate.
Six partly fused lips bearing very short labial papillae. Four distinct cephalic papillae.
Amphids indistinct. Stoma short and wide consisting from two cuticularized rings.
Oesophagus muscular with cylindrical procorpus, slightly swollen metacorpus a little
shorter than procorpus, short isthmus as wide as procorpus and basal bulb a bit wider
than metacorpus. Excretory pore past halfway from anterior to oesophageal base, 2–4 wi-
de. Cuticularized excretory duct 1–3 wide and 10–20 long. Excretory cell prominent.
Cardia conical. Intestine well developed, expanded posterior to oesophagus. Intestinal
walls 10–30 thick.
F i r s t g e n e r a t i o n f ema l e s. Cuticle with prominent broad annulations at
anterior. Stoma 9–11 wide and 4–5 deep. Oesophagus 181 (150–212) long. Procorpus
50–60 long and 20–25 wide, metacorpus 45–53 long and 31–34 wide, isthmus
18–25 long and 21–23 wide, basal bulb 50–60 long and 37–42 wide. Excretory pore at
90 (55–115) from anterior. Flexure of anterior ovary in 480-600 from oesophageal base.
Oviducts of four cells in cross-section. Mature oocytes 30–32 in diameter. Up to 16 gi-
ant amoeboid cells, round or elliptical in shape, 36–60 x 27–40 in size, in both uteri
and oviducts (fig. 1, D, E). Vulva at mid-body: V% = 52 (49–57). Vagina straight,
cuticularized, about 20 long. Vulva lips 3–5 long slightly protruded. Tail round or rarely
truncate, short (10–19) and wide (anal diameter 80–90). Small conical mucron 2–3 long.
S e cond g en e r a t i o n f ema l e s. Much shorter than first generation females.
Body C-shaped. Lips more prominent. Stoma 8–9 wide and 3–5 deep. Oesophagus
137–150 long. Procorpus 45–50 long and 15–17 wide, metacorpus 40–45 long and
22–25 wide, isthmus 15–20 long and 15–20 wide, basal bulb 30–40 long and 26–28 wi-
58 E. S. Ivanova, S. E. Spiridonov
de. Intestinal walls thinner than in first generation females. Anterior ovary flexure at
155–400 from oesophageal base. Eggs thin-shelled, 40–46x30 in size. Vulva lips pro-
truded. Up to 4 giant amoeboid cells in uteri. Tail broadly conical or rarely truncate,
longer (26–30) and narrower (anal diameter 35–55) than in first generation females.
Small mucron often presents.
59First report of Steinernema bicornutum...
Fig. 1. Morphology of Steinernema bicornutum from Krasnodar, Russia: À — 1st generation male, total view,
laterally; B — spicules and gubernaculum of type II, laterally; Ñ — male tail and spicules of type I, laterally;
D — giant amoeboid cell and smaller cell of similar structure; E — amoeboid cells in female oviduct; F —
infective juvenile, anterior end, laterally; G — infective juvenile, head end, ventrally; H — infective juvenile,
head end, laterally.
Ðèñ. 1. Ìîðôîëîãèÿ Steinernema bicornutum 1995 èç Êðàñíîäàðà, Ðîññèÿ: À — ñàìåö 1-ãî ïîêîëåíèÿ,
îáùèé âèä, ëàòåðàëüíî; B — ñïèêóëû è ðóëåê âòîðîãî òèïà, ëàòåðàëüíî; Ñ — õâîñò ñàìöà è ñïèêóëû
ïåðâîãî òèïà, ëàòåðàëüíî; D — ãèãàíòñêàÿ àìåáîèäíàÿ êëåòêà è íåáîëüøàÿ êëåòêà ñõîäíîé ñòðóê-
òóðû; E — àìåáîèäíûå êëåòêè â ÿéöåâîäå ñàìêè; F — ïåðåäíèé êîíåö èíâàçèîííîé ëè÷èíêè,
ëàòåðàëüíî; G — ãîëîâíîé êîíåö èíâàçèîííîé ëè÷èíêè, âåíòðàëüíî; H — ãîëîâíîé êîíåö èíâàçè-
îííîé ëè÷èíêè, ëàòåðàëüíî.
F i r s t g e n e r a t i o n ma l e s. Body J-shaped (fig. 1, A). Cuticle annulations not
visible. Stoma 6–8 wide and 3–5 deep. Oesophagus 145–161 long with procorpus 52–55
long and 12–14 wide, metacorpus 40–45 long and 17–21 wide, isthmus 23–25 long and
13–16 wide and basal bulb 27–35 long and 23–24 wide. Excretory pore 2–3 wide,
located in 90–110 from anterior, more distant than in females. Excretory cell less
prominent. Testis flexure at 550–680 from oesophageal base. Testis sometimes forms
one or two loops. Spicules (fig. 1, C) curved, yellow-brownish in colour. Capitulum well
separated from lamina, bowl-shaped, 13–18 long and 11–15 wide, calamus 9–12 wide
with protrusion, lamina curved 5–7 thick. Two cuticularized ridges 1–1.5 thick run
from calamus along lamina. Spicule tips straight, pointed, 2 thick with shallow trans-
verse incision. Velum 4–5 wide. Another type of spicules (fig. 1, B) occasionally seen:
Capitulum less separated from lamina and not much broader than latter one, spicule
tips a little wider. Gubernaculum spindle-shaped, flattened or convex, much variable in
size and direction of proximal end, which can be more or less separated, straight or
bent inwardly or outwardly. Muscles of tail end strongly developed. Six pairs of subven-
tral precloacal papillae and a single ventral adanal papillae, 3 pairs of subventral post-
cloacal papillae, 2 pairs near tail tip and 1 pair on dorsal side of tail tip (fig. 1, A, C).
No mucron presents.
S e c ond g en e r a t i o n ma l e s. Body thinner, without swelling at mid-part as in
first generation males. Oesophagus shorter and thinner (total length 127–130, width of
metacorpus 14–16, of bulb 19–21). Testis flexure at 280–330 from oesophageal base.
Spicules light yellow in colour. Velum thinner or absent. Both types of spicules repor-
ted above for first generation males present but type II more common.
I n f e c t i v e j u v en i l e s (fig. 1, F-H). Cuticle with distinct annulation. Two horn-
like structures on head end around mouth opening oriented in lateral position. Head
very slightly constricted at cervical region. Four prominent cephalic papillae in 1.5–2 back
60 E. S. Ivanova, S. E. Spiridonov
Fig. 2. The restriction patterns of Steinernema bicornutum from Krasnodar and Switzerland. Each pair of lanes
represents digestion of rDNA (about 1000 bp) of two cultures under comparison. Kr — S. bicornutum from
Krasnodar, Sw — S. bicornutum from Switzerland.
Ðèñ. 2. Ñïåêòðû ðåñòðèêöèè äëÿ Steinernema bicornutum èç Êðàñíîäàðà è Øâåéöàðèè. Êàæäàÿ ïàðà
äîðîæåê ïðåäñòàâëÿåò ðåçóëüòàò ðåñòðèêöèè ðèáîñîìàëüíîé ÄÍÊ ñðàâíèâàåìûõ êóëüòóð: Kr — S. bi-
cornutum èç Êðàñíîäàðà è Sw — S. bicornutum èç Øâåéöàðèè.
Kr Sw Kr Sw Kr Sw Kr Sw Kr Sw Kr Sw Kr Sw
Dde I Eco RI Cfo I IIind III IIind I Rsa I Pvu II
from head end. Stoma about 5–7 long, collapsed. Oesophagus thin, metacorpus very
slightly swollen (up to 7), isthmus long and thin, basal bulb 25–27 long and 10–12 wi-
de. Valve ridges of bulb poorly visible. Excretory pore 1 thick, cuticularized duct about
10 long, situated halfway from anterior to oesophageal base. Vesicle containing bacte-
rial cells distinct in few specimens, 12–30x5–17 in size in three-month-old juveniles.
Lateral field 6 wide.
Ana l y s i s o f rDNA s e qu en c e s. The RFLP comparison of type and Krasno-
dar populations of S. bicornutum revealed marked differences only in Cfo I profiles.
When type culture of S. bicornutum is characterized with the profile of two bands (i. e.
one restriction site for compared rDNA sequence), the nematodes from Krasnodar
population are characterized by three bands in Cfo I profile (fig. 2). The alignement of
partial rDNA sequences for both populations demonstrate molecular basis of observed
RFLP differences: two restriction sites for Cfo I were found in the sequence of Krasnodar
population, when only single Cfo I restriction site for type Yugoslavian culture (fig. 3).
Discussion
The data on morphometry of these two S. bicornutum isolates are presented in the
table 1. The values for type specimens of the species were obtained from original de-
scription (Tallosi et al., 1995). The mean value for spicule length is higher in Krasnodar
specimens than in Yugoslavian ones, though gubernaculum length ranges are quite close
for these two isolates. The ranges for spicule length are not overlapping in second gen-
eration males of both isolates. The mean length of infective juveniles from Krasnodar
was found to be lower than in type isolate (701 vs. 770), though the data for S. bicornu-
tum infective juveniles reported by D. Sturhan (personal communication) for juvenile
body length — 717(608–873) — are closer to those of Krasnodar isolate.
The restriction analysis of ribosomal DNA sequences of both S. bicornutum popula-
tions revealed difference in the profiles of Cfo I endonuclease. The comparison of
Ta b l e 1. Comparative morphometrics of Steinernema bicornutum of type culture and Krasnodar isolate,
Russia, mean value ± standard deviation (range)
Ò à á ëèö à 1. Ñðàâíèòåëüíûå äàííûå ïî ìîðôîìåòðèè òèïîâîé êóëüòóðû è êðàñíîäàðñêîãî èçîëÿòà S. bi-
cornutum
Character
Type specimens from Strazilovo,
Vojvodina, Yugoslavia (Tallosi,
Peters, Ehlers, 1995)
Specimens collected near Plant
Protection Institute, Krasnodar,
Russia
Body length of infective juvenile 770 ± 52 (648–873) 701 ± 94 (548–840)
Oesophagus length of infective
juveniles 124 ± 6 (113–135) 129 ± 8 (112–145)
Body length of first generation
males 1353 ± 150 (945–1539) 1530 ± 97 (1418–1675)
Distance between anterior end
and excretory pore of first genera-
tion males 82 ± 8 (68–98) 90 ± 15 (62–110)
Spicule length of first generation
males 65 ± 4.3 (53–70) 84 ± 9.6 (66–94)
Gubernaculum length of first ge-
neration males 48 ± 3.5 (38–50) 44 ± 1.9 (40–46)
Spicule length of second generati-
on males 51 ± 1.5 (48–53) 71 ± 9.7 (59–82)
Gubernaculum length of second
generation males 33 ± 2.9 (25–38) 37 ± 2.8 (31–40)
61First report of Steinernema bicornutum...
62 E. S. Ivanova, S. E. Spiridonov
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715 bp long sequence of rDNA of Krasnodar isolate with the similar sequence of Yugo-
slavian S. bicornutum (Nguyen et al., 2001) revealed the molecular basis for observed
RFLP difference: because of the single nucleotide C-A substitution (fig. 3) the nema-
todes of type culture lack second Cfo I restriction site. Totally differences in 18 pairs
of nucleotides were found between these two isolates including 17 substitutions and one
indel in Yugoslavian strain. Thus, 2.2% of nucleotides from this 715 bp part of riboso-
mal DNA are different in the isolates under comparison. We have found the difference
in 2.4% of rDNA nucleotides between the populations of another steinernematid spe-
cies — S. feltiae. The fertile crosses between the steinernematid populations with such
level of nucelotide differences were obtained (own unpublished observations). Much
higher differences (more than in 15% of nucleotides) were found between S. bicornu-
tum and the closest species of the genus — S. ceratophorum, which was described from
China (Jian et al., 1997; A. Reid, personal communication, unpublished). The absence
of clear morphological differences between studied isolates persuades us to consider
these as conspecific. This finding is the first report of S. bicornutum for the territory of
former USSR. Our results indicate that European S. bicornutum is splad at least up to
northern slope of the Caucasus. Infective juveniles of S. bicornutum from Krasnodar
were found to be highly invasive for Galleria mellonella caterpillars. Both these nemato-
des and their symbiotic bacteria represent promising object for the research in biocontrol.
The work was supported by Russian Foundation for Basic researches (grant 02–04–48389). The authors
are grateful to Dr. Oksana Ivakhnenko for her help and guidance in the gardens of Krasnodar Institute of
Plant protection where nematodes were collected, to Dr. Igor Kramer and Dr. Jurg Grunder (Federal Agri-
cultural Research Centre in Wadenswil, Switzerland) for lending the Swiss culture of S. bicornutum and to
Dr. Maurice Moens (CLO, Merelbeke, Belgium) for his support of EPN research in Russia, and for his per-
mit to use his laboratory resources to sequence steinernematids from Krasnodar.
Hominick W. M., Briscoe B. R., del Pino F. G. et al. Biosystematics of entomopathogenic nematodes: current
status, protocols and definitions. // J. Helminthology. — 1997. — 71. — P. 271–298.
Jian H., Reid A. P., Hunt D. J. Steinernema ceratophorum n. sp. (Nematoda, Steinernematidae), a new
entomopathogenic nematode from north-east China. // Systematic Parasitology. — 1997. — 37, N 1.
— P. 115–125.
Joyce S. A., Reid A., Driver F., Curran J. Application of polymerase chain reaction (PCR) A. M., R. — U.
Ehlers and J. P. Masson, Eds. COST 812 Biotechnology: Genetics of entomo-pathogenic nematodes-
bacterium complexes // Proceedings of symposium and workshop, S. Patrick's college, Maynooth. CO.
Kildare, Ireland. EC DG XII: Luxembourg. 1994. — P. 178–187.
Nguyen K. B., Maruniak J., Adams B. J. Diagnostic and phylogenetic utility of the rDNA internal transcribed
spacer sequences of Steinernema // J. Nematology. — 2001. — 33, N 2–3. — P. 73–82.
Tallosi, B., Peters A., Ehlers R.-U. Steinernema bicornutum sp. n. (Rhabditida: Steinernematidae) from
Vojvodina, Yugoslavia // Russian Journal of Nematology. — 1995. — 3, N 2. — P. 71–80.
Vrain T. C., Wakarchuk D. A., Levesque A. C., Hamilton R. T. Intraspecific rDNA restriction fragment length
polymorphisms in the Xiphinema americanum group // Fundamental and Applied Nematology. —
1992. — 15. — P. 567–574.
63First report of Steinernema bicornutum...
|
| id | nasplib_isofts_kiev_ua-123456789-3837 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0084-5604 |
| language | English |
| last_indexed | 2025-12-07T16:25:26Z |
| publishDate | 2003 |
| publisher | Інститут зоології ім. І. І. Шмальгаузена НАН України |
| record_format | dspace |
| spelling | Ivanova, E.S. Spiridonov, S.E. 2009-07-10T12:22:44Z 2009-07-10T12:22:44Z 2003 First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Caucasus / E. S. Ivanova, S. E. Spiridonov // Вестн. зоологии. — 2003. — Т. 37, № 3. — С. 57-63. — англ. 0084-5604 https://nasplib.isofts.kiev.ua/handle/123456789/3837 576.895.132 Нематоды Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 были выделены, с использованием гусениц Galleria mellonella в качестве приманок, из образцов почвы, собранных около Краснодара (Северный Кавказ). Обнаружены различия между краснодарской и типовой (югославской) культурами S. bicornutum в средней длине инвазионных личинок и длине спикул самцов. Все эти различия (за исключением длины спикул у самцов второй генерации) не являются статистически достоверными. Рестрикционный анализ рибосомальной ДНК дифференцирует эти две популяции S. bicornutum лишь по спектрам эндонуклеазы Cfo I (один сайт рестрикции у типовой, и два — у краснодарской культуры). Проведено секвенирование 715 пар оснований фрагмента рибосомального гена (ITS1 + 5.8S + ITS2). Сравнение с последовательностью типового изолята показало различие в 18 парах нуклеотидов, включая 17 замен и одну делецию в типовой культуре. Выявленные морфологические и молекулярные различия оцениваются как межпопуляционные, а обнаруженные в Краснодаре нематоды как еще один изолят S. bicornutum. Steinernema bicornutum Tallosi, Peters & Ehlers, 1995 was isolated by Galleria baiting from soil samples collected near Krasnodar (North Caucasus). The differences were found between the type (Yugoslavian) population and Krasnodar isolate in the average length of infective juveniles and spicules. All these differences (except spicule length in the second generation males) are not statistically significant. The RFLP analysis was able to distinguish nematodes from these two populations of S. bicornutum only in the Cfo I profiles (one restriction site in the type culture and two restriction sites in the Krasnodar isolate). The 715 bp long fragment of ribosomal gene (ITS1 + 5.8S + ITS2) of the Krasnodar isolate of S. bicornutum was sequenced and compared with similar sequence of the type strain. In accordance with RFLP data, one additional Cfo I restriction site was found in the Krasnodar S. bicornutum. Totally, the difference in 18 pairs of nucleotides per fragment of 715 bp was found between Krasnodar and type cultures, including 17 substitutions and one deletion (in type culture). Reported morphological and molecular differences are considered as interpopulational ones, and described nematodes from Krasnodar as another isolate of S. bicornutum. en Інститут зоології ім. І. І. Шмальгаузена НАН України Краткие сообщения First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus Первая находка Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) на Северном Кавказе Article published earlier |
| spellingShingle | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus Ivanova, E.S. Spiridonov, S.E. Краткие сообщения |
| title | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus |
| title_alt | Первая находка Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) на Северном Кавказе |
| title_full | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus |
| title_fullStr | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus |
| title_full_unstemmed | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus |
| title_short | First report of Steinernema bicornutum (Nematoda, Rhabditida, Steinernematidae) from the North Cau-casus |
| title_sort | first report of steinernema bicornutum (nematoda, rhabditida, steinernematidae) from the north cau-casus |
| topic | Краткие сообщения |
| topic_facet | Краткие сообщения |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/3837 |
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