Rapid identification of <i>Henosepilachna vigintioctopunctata</i> and <i>Henosepilachna vigintioctomaculata</i> based on species-specific mitochondrial cytochrome oxidase I primers

GUO Mujuan; LIN Miaojin; PAN Guang; YU Jiaqi; LÜ Jing; GUO Wei; ZHANG Yafeng; YANG Chunxiao; QIU Baoli; ZHOU Xuguo; PAN Huipeng

doi:10.7671/j.issn.1001-411X.202101008

Journal of South China Agricultural University > 2022 > 43(1): 59-66. > DOI: 10.7671/j.issn.1001-411X.202101008

GUO Mujuan, LIN Miaojin, PAN Guang, et al. Rapid identification of Henosepilachna vigintioctopunctata and Henosepilachna vigintioctomaculata based on species-specific mitochondrial cytochrome oxidase I primers[J]. Journal of South China Agricultural University, 2022, 43(1): 59-66. DOI: 10.7671/j.issn.1001-411X.202101008

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Rapid identification of Henosepilachna vigintioctopunctata and Henosepilachna vigintioctomaculata based on species-specific mitochondrial cytochrome oxidase I primers

1.
College of Plant Protection, South China Agricultural University/Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, Guangzhou 510642, China
2.
National Modern Agricultural Industry Science and Technology Innovation Center of Guangzhou, Guangzhou 510520, China
3.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, South China Agricultural University, Guangzhou 510642, China
4.
Department of Entomology, University of Kentucky, Lexington 40546, USA

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Corresponding author:
PAN Huipeng
Received Date: January 05, 2020
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
Henosepilachna vigintioctopunctata (Fabricius) and Henosepilachna vigintioctomaculata (Motschulsky), two kinds of phytophagous ladybeetles in China, are destructive pests causing great damage to solanaceous plants. They are difficult to distinguish based on external morphological characteristics, and it is therefore necessary to develop a rapid and accurate method to differentiate them.

Method
We established a molecular identification technique using species-specific (SS) PCR primers based on the species-specificity of mitochondrial cytochrome oxidase I (mtCOI) of two ladybeetles. Two pairs of SS-mtCOI primers, Hvp and Hvm, were designed based on sequence variations in the mtCOI gene between H. vigintioctopunctata and H. vigintioctomaculata.

Result
PCR amplifications were conducted using these two primers and both had species-specific amplifications. Sensitivity assays were conducted under different DNA concentrations, and the results showed that Hvp primers for H. vigintioctopunctata had a detectable amplification band at a DNA concentration of 3.13 mg/L, while Hvm primers for H. vigintioctomaculata had a detectable amplification band at a DNA concentration of 2.43 mg/L. The egg or 1st instar of H. vigintioctopunctata and H. vigintioctomaculata could also be accurately differentiated by Hvp and Hvm primers. Furthermore, six field populations of H. vigintioctopunctata collected from six provinces could be authenticated by the Hvp primers.

Conclusion
These two pairs of SS-mtCOI primers can differentiate H. vigintioctopunctata and H. vigintioctomaculata rapidly, accurately and sensitively.

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References (26)

References

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Rapid identification of Henosepilachna vigintioctopunctata and Henosepilachna vigintioctomaculata based on species-specific mitochondrial cytochrome oxidase I primers

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