Research Article |
Corresponding author: Narin Chomphuphuang ( narich@kku.ac.th ) Corresponding author: Kittisak Kumtanom ( kittisak.k@pit.ac.th ) Academic editor: J. Adilson Pinedo-Escatel
© 2025 Kanyakorn Piraonapicha, Nithina Kaewtongkum, Narin Chomphuphuang, Panrak Kimsawat, Kittisak Kumtanom, Yudthana Samung.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Piraonapicha K, Kaewtongkum N, Chomphuphuang N, Kimsawat P, Kumtanom K, Samung Y (2025) Mukaria sakaeratensis sp. nov. (Hemiptera, Cicadellidae, Deltocephalinae), a new species of bamboo leafhopper from Sakaerat Biosphere Reserve, Thailand. ZooKeys 1239: 305-320. https://doi.org/10.3897/zookeys.1239.145803
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Mukaria sakaeratensis Piraonapicha & Chomphuphuang, sp. nov. is described based on male and female specimens recently collected in Nakhon Ratchasima, Thailand. The new species is herein described by an integrative approach combining morphological and molecular evidence. Genetic distance analyses revealed a potential barcoding gap (K2P) of 0.20–12.07% for COI in Mukaria. Species delimitation methods ABGD and ASAP demonstrated promising results for the COI gene. This species clearly differs from all its congeners in the aedeagal shaft abruptly narrowed and curved inward in the distal half, and with a pair of spines pointed anteriorly. Mukaria sakaeratensis sp. nov. has been found on the bamboo Vietnamosasa pusilla (A. Chev. & A. Camus) T.Q. Nguyen. This finding constitutes the first recorded instance of a specialized member of the tribe Mukariini (Hemiptera: Cicadellidae: Deltocephalinae) feeding exclusively on bamboo from the genus Vietnamosasa. The holotype has been deposited in the Entomology Section, Queen Sirikit Botanic Garden, The Botanical Garden Organization, Thailand.
Barcoding gene, COI, deciduous dipterocarp forests, identification key, molecular identification, morphology, Mukariini, Thailand, Vietnamosasa
The leafhopper genus Mukaria was established by Distant in 1908 based on the type species Mukaria penthimioides from Sri Lanka. Currently, the genus consists of 15 valid described species reported in Bangladesh, China, India, Indonesia, Japan, Sri Lanka, Thailand, and Pakistan (
In this study, we describe a new species, Mukaria sakaeratensis sp. nov., discovered at the Sakaerat Environmental Research Station within the Sakaerat Biosphere Reserve, Nakhon Ratchasima Province, in northeastern Thailand. This species is characterized by its dark-colored body and was found exclusively on its host plant, the small-sized sympodial bamboo Vietnamosasa pusilla (A.Chev. & A.Camus) T.Q.Nguyen, in dry dipterocarp forests. The description of Mukaria sakaeratensis sp. nov. is based on an integrative approach, combining detailed morphological analyses of both male and female specimens with molecular evidence derived from COI barcoding, which provides robust DNA-based support for its classification.
The Mukaria specimens were collected from a restoration forest within deciduous dipterocarp forests, Sakaerat Biosphere Reserve, Nakhon Ratchasima Province, Thailand. The new species was found on leaves of Vietnamosasa pusilla (A.Chev. & A.Camus). Specimens of the new species were prepared through dry pinning and preservation in alcohol for morphological examination. The genitalia of males and females were dissected from the abdominal segment. The dissected genitalia were cleared in a 10% potassium hydroxide (KOH) solution for one day at room temperature, washed with distilled water, and then stored in glycerine in microvials before examination and imaging. Morphological observations were conducted using a Nikon SMZ445 stereomicroscope. Images were taken using a Nikon Digital Sight Ri1 camera attached to a Nikon AZ100M stereomicroscope and processed with NIS-Elements-D for a multi-focused montage. Images of genitalia and wing venation were taken using a Nikon DS-F12 camera attached to a Nikon Eclipse Ci-L compound microscope. Images of living workers and nest entrances were taken using a Canon RF100 mm f/2.8L macro lens attached to a Canon R6 digital camera. Specimens are deposited in the Entomology Section, Queen Sirikit Botanic Garden, The Botanical Garden Organization, Chiang Mai Province, Thailand (
Tissue samples from the right legs of male and female adults were preserved in 95% ethanol. Total DNA was extracted following the DNeasy Blood & Tissue Kit (Qiagen) protocol, and the extracted DNA was stored at -20 °C. The PCR reaction mix had a total volume of 50 μl, comprising 20 μl ultrapure water, 3 μl of the DNA template, 1 μl of each primer (10 μM), and 25 μl of master mix. Thermal cycling started with incubation at 94 °C for 1 min, followed by 40 cycles at 94 °C for 30 s, annealing at 48, 50 °C for 50 s, and extension at 72 °C for 1 min, with a final extension step at 72 °C for 5 min. All PCR products were visualized on 1.5% agarose gels using Omnipur Agarose (United States of America). A 592 bp fragment of the COI gene was amplified using universal primers LCO1490 and HCO2198, as described by
Purified PCR products underwent bidirectional Sanger sequencing on the ABI PRISM 3130x.1 Genetic Analyzer (Applied Biosystems, Foster City, USA) Sequencing was performed by Macrogen Inc. Sequencing (Seoul, Korea). Consensus sequences were generated, and sequences of the candidate DNA barcodes aligned using ClustalW and verified using BIOEDIT v. 7.2.5 (
Species | Locality | Sex | Voucher | Accession number/ sequence ID | Sources |
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M. sakaeratensis Piraonapicha & Chomphuphuang, sp. nov. | Nakhon Ratchasima, Thailand | Female |
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PP582202 | This study |
Nakhon Ratchasima, Thailand | Female |
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PP582203 | This study | |
Nakhon Ratchasima, Thailand | Female |
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PP582204 | This study | |
Nakhon Ratchasima, Thailand | Female |
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PP582205 | This study | |
Nakhon Ratchasima, Thailand | Male |
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PP582206 | This study | |
Nakhon Ratchasima, Thailand | Male |
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PP582207 | This study | |
Nakhon Ratchasima, Thailand | Male |
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PP582208 | This study | |
Nakhon Ratchasima, Thailand | Male |
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PP582209 | This study | |
M. splendida | Yunnan, China | Male | MG813485 |
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MK862276 | GenBank | ||||
MW487892 | GenBank | ||||
India | Male | OM869458 |
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India | Male | OM345004 |
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India | Male | OP617462 |
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India | Male | OP616038 |
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Punjab, Pakistan | GMPJA9077-21.COI-5P | BOLD Systems | |||
Punjab, Pakistan | GMPJA609-21.COI-5P | BOLD Systems | |||
Punjab, Pakistan | GMPJA5825-21.COI-5P | BOLD Systems | |||
Punjab, Pakistan | GMPJA4951-21.COI-5P | BOLD Systems | |||
KwaZulu-Natal, South Africa | SAKZA3134-22.COI-5P | BOLD Systems | |||
Chittagong, Bangladesh | GMBDE4119-23.COI-5P | BOLD Systems | |||
M. maculata | MG736687 | GenBank | |||
MG736688 | GenBank | ||||
MG736689 | GenBank | ||||
MG736690 | GenBank | ||||
M. albinotata | MG736685 | GenBank | |||
MG736686 | GenBank | ||||
M. bambusana | MG736694 | GenBank | |||
MG736695 | GenBank | ||||
MG736696 | GenBank | ||||
MG736697 | GenBank | ||||
Deltocephalus vulgaris | Hainan, China | MT998308 |
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To assess phylogenetic relationships, COI sequences from previous studies were retrieved from GenBank, as detailed in Table
Two methods were used for phylogenetic analysis: maximum likelihood (ML) and Bayesian inference (BI). The ML reconstructions were performed using RAxML-NG (
Mukaria Distant, 1908: 269. Type species: Mukaria penthimioides Distant, 1908, by original designation.
Parabolotettix
Matsumura, 1912: 280.Type-species: Parabolotettix maculatus Matsumura, 1912, by original designation. Synonymised by
Ikomella
Ishihara, 1961: 253. Type species: Ikomella confersa Ishihara, 1961, by original designation. Synonymised by
Revised from
M. albinotata Cai & Ge, 1996: China (
M. creagra Zhao, Luo & Chen, 2024: China (
M. confersa (Ishihara, 1961): Thailand (
M. flavida Cai & Ge, 1996: China (
M. hainanensis Yao, Yang & Chen, 2019: China (
M. lii Yang & Chen, 2011: China (
M. maculata (Matsumura, 1912): China, Japan and Indonesia (
M. nigra Kuoh & Kuoh, 1983: China (
M. omani Viraktamath & Webb, 2019: India (
M. penthimioides Distant, 1908: Sri Lanka and India (
M. splendida Distant, 1908: Bangladesh, India and Pakistan (
M. striola Zhao, Luo & Chen, 2024: China (
M. vakra Viraktamath & Webb, 2019: India (
M. variabilis Evans, 1973: Indonesia (New Guinea) (
M. zonata Hayashi, 1996: Japan (
Thailand. Holotype • One male, Sakaerat Biosphere Reserve, Udomsub Sub-District, Wang Nam Khiao District, Nakhon Ratchasima Province, Thailand, 14°30'33.7"N, 101°56'23.4"E, 326 m a.s.l., 10.VI.2024, N. Kaewtongkum leg. (
Measurements. Male. Body length (including tegmen) 2.92 ± 0.12 mm (N = 10); head widths 0.87 ± 0.02 mm (N = 10). Female. Body length (including tegmen) 3.07 ± 0.11 mm (N = 10); head widths 0.92 ± 0.03 mm (N = 7).
Coloration. Male. Head (excluding eyes) entirely black in dorsal view, postclypeus dark brown with median area brown; lorum and gena dark brown; pronotum and scutellum dark brown; forewing dark brown, apical 1/4 with brown, yellow spot at mid-length extending to lateral margin of pronotum, oblique spot on costa, subtriangular spot near outer apical cell, small yellow spot near ScP+RA, hindwing brown, hyaline; wing veins brown; coxa and trochanter of all legs dark brown, femur, tibia and tarsi of fore and middle leg yellow; femur and tibia of hindleg brown, apex of tibia dark brown; basal half of 1 segmented tarsi segment pale brown, apical half dark brown, 2 segmented of tarsi pale brown, 3 segmented of tarsi and claw dark brown. Female. Similar to male.
Male genitalia.
In lateral view, male pygofer subtriangular, approximately twice as length as high. In ventral view, valve subtriangular and wider than long and with anterior margin slightly concave and posterior margin produced medially. In ventral view, subgenital plate subrounded, with subtriangular shaped apex, inner margin roundly convex, outer margin almost straight, slightly concave near apex, basal part of subgenital plate with approximately 12 long setae, and distal part with 9–13 long setae. Style, slightly wider at base with lateral process, preapical lobe rectangular, strongly concave near apex. Apex of style digitiform and curved outward. Connective Y shaped, stem as long as arms. Aedeagus with a pair of arcuate laterobasal processes, nearly 1/3 as long as aedeagal shaft, and aedeagal shaft abruptly narrowed and curved inward in distal half, half portion in ventral view with a pair of spines pointed anteriorly. In lateral view, near apex of aedeagus with strong subtriangular lobe pointed anteriorly (Fig.
Female genitalia.
Female sternite VII subrectangular, approximately 2 times longer than wide, with pair subtriangular lobes at apex, outer margin convex and strongly emarginate in middle. Valvula I almost straight. Valvula II similar to valvula I, but 1/3-part upper margin serrate. Pygofer spinose in posterior half, ovipositor not exceeding pygofer (Fig.
Thailand (Nakhon Ratchasima Province) (Fig.
The specific epithet ‘sakaeratensis’ refers to the type locality.
The bamboo species Vietnamosasa pusilla (A.Chev. & A.Camus) T.Q. Nguyen has been identified as the host plant for Mukaria sakaeratensis sp. nov., a newly described species of bamboo-feeding leafhopper discovered in the Sakaerat Biosphere Reserve, Thailand. This finding marks the first documented ecological relationship between the bamboo genus Vietnamosasa and members of the tribe Mukariini (Hemiptera: Cicadellidae: Deltocephalinae), which are specialized herbivores feeding exclusively on bamboos. The genus Vietnamosasa comprises sympodial bamboos found in Southeast Asia, including Thailand, Cambodia, Laos, and Vietnam. Vietnamosasa pusilla, commonly found in dry dipterocarp forests, is characterized by its small size and adaptation to fire-prone environments. During the collection of the new species of Mukaria, the plant height, measured from the base of the stem (at the soil surface) to the tallest part of the plant, was approximately 70–90 cm.
In this study, genetic distances were calculated for the cytochrome c oxidase subunit I (COI) gene across several Mukaria species. The intraspecific genetic distances ranged from 0.10% to 0.20%, indicating relatively low genetic variation within individual species. When comparing between species, the interspecific genetic distances varied considerably, with the highest value of 23.71% observed between M. bambusana and M. maculata (2). Conversely, the lowest interspecific genetic distance was 12.07%, found between M. sakaeratensis sp. nov. and M. splendida (Fig.
The genus Mukaria is distributed in the southeastern Palaearctic, Oriental and Oceanic regions. Three species were reported in Southeast Asia, i.e., M. confersa, M. maculata and M. variabilis. Prior to this study, there was a long gap of 63 years during which no new species of the genus were reported from Thailand.
Mukaria sakaeratensis Piraonapicha & Chomphuphuang, sp. nov., male, A–C genital capsule, ventral view (A), dorsal view (B), lateral view (C), D valve and subgenital plate in ventral view E–G style, connective and aedeagus, ventral view (E) dorsal view (F) lateral view (G). Scale bars: 0.2 mm (A–C), 0.1 mm (E–G).
Mukaria sakaeratensis Piraonapicha & Chomphuphuang, sp. nov., female genitalia, A, B genital capsule (A), lateral view (B), ventral view, C sternite VII D valvula I apex magnified in lateral view E valvula I in lateral view F valvula II, apex magnified in lateral view G valvula II in lateral view.
Molecular species delimitation of Mukaria using maximum likelihood and partial COI sequences. The tree was inferred from 592 base pairs of six species of Mukaria and one species of Deltocephalus as the outgroup. Nodal support values are bootstrap values (percentage of 1000 replicates). Node numbers represent two support values: bootstrap support from RAxML and posterior probability from Bayesian inference. The bars illustrate molecular delimitation methods, encompassing genetic distances (ABGD, ASAP). Scale bar indicates 0.050 nucleotide substitutions.
1 | Apex of aedeagus bifurcated | 2 |
– | Apex of aedeagus not bifurcated (Fig. |
M. sakaeratensis sp. nov. |
2 | Pair of spines near apex of aedeagus curved inward ( |
M. maculata |
– | Pair of spines near apex of aedeagus curved outward ( |
M. variabilis |
We sincerely thank Surachit Waengsothorn (Sakaerat Environmental Research Station, Thailand Institute of Scientific and Technological Research, Thailand) and Weeyawat Jaitrong (National Science Museum, Thailand) for their invaluable contributions to the discovery of the new species in Sakaerat Environmental Research Station. KP extends gratitude to Ji-Chun Xing (Guizhou University) for editing the manuscript. We also extend our thanks to Yuranan Nanthaisong, Piyarat Thawongsa, Benjapon Kaewwanna, Channarong Imrattanakul and Charanchai Chanthakhat for their fieldwork assistance and for taking photos of leafhoppers in the field.
The authors have declared that no competing interests exist.
This research has been reviewed and approved by the Institutional Animal Care and Use Committee of Khon Kaen University based on the Ethic of Animal Experimentation of the National Research Council of Thailand (Record No. IACUC-
This research was supported by Khon Kaen University has received funding support from the National Science Research and Innovation Fund (NSRF).
Kanyakorn Piraonapicha: Data curation (lead); formal analysis (lead); investigation (lead); methodology (lead); validation (lead); visualization (lead); writing – original draft (lead); writing–review and editing (equal). Nithina Kaewtongkum: Writing – review and editing (equal). Narin Chomphuphuang: Conceptualization (equal); methodology (equal); supervision (lead); funding acquisition (lead); writing – review and editing (equal). Panrak Kimsawat: methodology (equal). Kittisak Kumtanom: funding acquisition (equal); writing – review and editing (equal); methodology (equal); writing – review and editing (equal). Yudthana Samung: methodology (equal); visualization (equal).
Kanyakorn Piraonapicha https://orcid.org/0000-0003-3800-8625
Nithina Kaewtongkum https://orcid.org/0009-0001-8782-3055
Narin Chomphuphuang https://orcid.org/0000-0003-0738-3879
Panrak Kimsawat https://orcid.org/0009-0008-7743-8947
Kittisak Kumtanom https://orcid.org/0009-0000-9059-7872
Yudthana Samung https://orcid.org/0000-0003-4069-4257
All of the data that support the findings of this study are available in the main text.