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THE MORPHOMETRIC CHARACTERISTICS AND MOLECULAR-
GENETIC ANALYSIS OF ORYCTES NASICORNIS (LINNAEUS, 1758)
DISTRIBUTED IN THE LOW HILLS OF KARAKALPAKSTAN
Amangul Sarsenbaevna Yusupova
Independent Researcher,
Institute of Zoology,
Academy of Sciences of the Republic of Uzbekistan
https://doi.org/10.5281/zenodo.14863375
Abstract.
This article presents an analysis of the morphometric
characteristics and molecular-genetic results of Oryctes nasicornis (Linnaeus,
1758) distributed in the low hills of Karakalpakstan. Significant differences in
the morphological structure between male and female representatives of
Oryctes nasicornis were clearly identified. Although morphological identification
remains one of the most widely used and generally accepted methods, it may
lead to misidentifications due to intraspecific morphological variations. No
nucleotide differences were observed within Oryctes nasicornis. However,
molecular-genetic analysis revealed a 15.6% nucleotide divergence between
Oryctes nasicornis and Oryctes rhinoceros based on sequences obtained from
the GenBank database.
Keywords:
Low hills, morphometric characteristics, head structure, genus,
species.
Introduction
The considerable size of
Oryctes nasicornis
(Linnaeus, 1758) becomes
evident when it unexpectedly collides with us while flying in the evening,
allowing us to physically sense its substantial weight. To visualize the strength
or size of insects, we can compare them to other organisms: for instance, an
elephant can lift 30–50% of its div weight, while a human can lift 50–80% of
their own weight. If a human weighing approximately 70 kg possessed the
strength of
Oryctes nasicornis
, they would be able to move several railway
wagons weighing up to 7 tons with ease. Given that beetles can support multiple
times their div mass, developed countries are actively conducting research on
remotely photographing such biological objects. Additionally, there is ongoing
exploration into the potential of radio control over these insects during flight.
The morphological structure of
Oryctes nasicornis
varies significantly
between male and female individuals. The div length of males ranges from 25–
43 mm, reaching up to 47 mm, while in females, it ranges from 26–41 mm. The
beetle is large, with a dark brown-reddish or reddish-brown coloration. The
ventral part of the div and femora are relatively lighter, appearing yellowish-
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reddish-brown. The div is moderately elongated and convex, with a noticeably
broad structure. The head, pronotum, and especially the elytra are highly glossy.
The head is relatively small, with a triangular clypeus that narrows slightly
at the front. In males, the clypeus is covered with fine, non-fused punctures,
whereas in females, it is densely covered with deep punctures and wrinkles. The
eyes are wide, and the antennae are fan-shaped with a large basal segment. The
upper mandibles are broad, lacking external teeth. In males, the majority of the
head is occupied by a large horn, which gradually narrows. The anterior surface
of the horn is flat, lacking a longitudinal groove in the middle, and is densely
covered with fine punctures. The posterior part of the head is smooth. In
females, the horn is much smaller and often forms an angle with the anterior
part of the head.
The pronotum is slightly transverse, with the anterior part bending
downward and gradually rising towards the back. The anterior margin of the
pronotum features a deep triangular depression bordered by a wide, smooth
rim. The anterior angles of the pronotum are sharp, with rounded tips that
protrude forward, while the posterior angles are rounded (Figure 1). Both sexes
exhibit dense, small punctures on the thoracic region. The legs are robust but
not particularly long, with the hind legs being the longest and slightly thickened.
The middle and hind tibiae gradually widen towards the apex and possess long
teeth at the tip. The coxae of the hind legs are significantly thickened.
Figure 1.
Molecular-genetic analysis of
Oryctes nasicornis
(Linnaeus, 1758).
Morphological identification is currently one of the most widely used and
generally accepted methods. However, intraspecific morphological variations
may lead to misidentification (St Laurent B, 2016; St Laurent B & Supratman S,
2016; Lobo NF, 2015). One of the challenges associated with morphological
identification is the damage to diagnostic characteristics, which can make it
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difficult or even impossible to identify species with similar or insufficiently
distinct traits (Lobo NF, 2015). Moreover, accurate morphological identification
requires thorough and meticulous preparation.
Molecular identification offers more precise results and can be particularly
useful in biodiversity-rich regions where numerous closely related species exist
(Davidson JR, 2020; Sinka ME, 2011; Stresman GH, 2010). Mitochondrial genome
structure and evolution provide substantial data for genetic and phylogenetic
population studies. Currently, specific mitochondrial genome genes are widely
used to analyze interspecies and intraspecies differences. For instance, the COI
gene sequence has been employed for mosquito species identification through
mtDNA barcoding (Gao B & Fang Y, 2012; Wang G & Li C, 2012). Additionally,
genes such as SOI (Feng X, 2017), COII (Yang M & Ma Y, 2015), and ND5
(Makhawi AM, 2013) have been utilized as markers for assessing genetic
population structure.
The objective of this study is to perform the molecular genetic classification
of
Oryctes nasicornis
(Linnaeus, 1758), collected from the low hill regions of
Karakalpakstan, which belongs to the genus
Oryctes
Hellwig, 1798.
Molecular genetic analyses of
Oryctes
species were conducted based on
sequencing chromatography. The mitochondrial DNA (mtDNA) of
Oryctes
nasicornis
was extracted, focusing on the COI region, consisting of 556 base
pairs. To compare these sequences, reference nucleotide data were obtained
from
the
International
Bioinformatics
Information
Center
https://blast.ncbi.nlm.nih.gov
), specifically for
Oryctes nasicornis
(Accession
Number: MZ633039) and
Oryctes rhinoceros
(Accession Number: KY313828).
According to bioinformatic analysis, no nucleotide differences were found
between the studied
Oryctes nasicornis
specimens and the reference
Oryctes
nasicornis
sequence from the GenBank database (as shown in Figure 2).
Figure 2.
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Comparative Analysis of
Oryctes nasicornis
and
Oryctes rhinoceros
Nucleotide Sequences. A comparative analysis of nucleotide sequences was
conducted for
Oryctes nasicornis
and
Oryctes rhinoceros
, both belonging to the
genus
Oryctes
Hellwig, 1798. The nucleotide differences between
Oryctes
nasicornis
and
Oryctes rhinoceros
(GenBank Accession Number: KY313828)
were identified at 103 positions. Specifically:
At positions
13, 14, 118, 145, 302, 343, 364, 385, 412, 442, 445,
475, 521, 526, 532, 547, and 581
,
Oryctes nasicornis
had thymine (T),
whereas
Oryctes rhinoceros
had cytosine (S).
At positions
22, 88, 274, 349, and 574
,
Oryctes nasicornis
had
adenine (A), while
Oryctes rhinoceros
had cytosine (S).
At positions
28, 58, 73, and 433
,
Oryctes nasicornis
had adenine (A),
whereas
Oryctes rhinoceros
had guanine (G).
At positions
31, 43, 223, 280, 319, 373, 376, 400, 477, 550, 562,
568, 592, 631, and 637
,
Oryctes nasicornis
contained thymine (T), while
Oryctes rhinoceros
had adenine (A).
At positions
40, 52, 70, 171, 205, 286, 337, and 541
,
Oryctes
nasicornis
had cytosine (S), whereas
Oryctes rhinoceros
had adenine (A).
At positions
50, 106, 136, 427, 490, 496, 505, 514, 529, 535, 571,
601, 610, 616, and 646
,
Oryctes nasicornis
had cytosine (S), whereas
Oryctes rhinoceros
had thymine (T).
The molecular-genetic comparison also revealed that
Oryctes nasicornis
and
Oryctes rhinoceros
(GenBank Accession Number: KR143134) exhibited
15.6%
nucleotide sequence divergence
.
Phylogenetic tree. Based on the results of molecular-genetic research, the
analysis of nucleotide sequences of the COI region of mitochondrial DNA from
the studied species of the genus Oryctes and those obtained from the GenBank
database revealed that representatives of this genus are grouped into two
clades. (Figure 3).
Figure 3.
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A phylogenetic family tree developed on the basis of the maximum
likelihood (ML) method of mites of the genus Oryctes. Oryctes rhinoceros and
Oryctes nasicornis species were placed in the first group, and representatives of
this species united to form a butistrap support of 97-99%. In the second group,
the Oryctes nasicornis species has 82% relative to the main joint, and 99%
within the species, forming a butistrap support.
Thus, based on the results of the molecular genetic research conducted on
the species Oryctes nasicornis nasicornis belonging to the genus Oryctes, the
nucleotide sequences belonging to the COI region of the mRNA were placed in
the Genbank database and accession numbers were obtained.
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