HcAnk1 and HcAnk2 genes function and HcNPV susceptibility of Hyphantria cunea based on RNAi

doi:10.12302/j.issn.1000-2006.202203049

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2024, Vol. 48 ›› Issue (3): 181-190.doi: 10.12302/j.issn.1000-2006.202203049

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HcAnk1 and HcAnk2 genes function and HcNPV susceptibility of Hyphantria cunea based on RNAi

PENG Mengmeng¹(), WU Hongqu¹^,²(), ZHANG Jiawen¹, YAN Liqiong¹, CAO Chuanwang¹, SUN Lili¹^,^*()

1. Key Laboratory of Sustainable Forest Ecosystem Management-Ministry of Education, Northeast Forestry University, Harbin 150040, China
2. Hubei Biopesticide Engineering Research Center, Wuhan 430070, China

Received:2022-03-24 Revised:2022-09-05 Online:2024-05-30 Published:2024-06-14
Supported by:
National Natural Science Foundation of China(32171791);National College Students’ Innovation and Entrepreneurship Training Program(202010225031);Natural Science Foundation of Heilongjiang(YQ2021C007)

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Abstract

Abstract:

【Objective】 Ankyrin is a family of adaptor proteins that connect the cytoskeleton to the plasma membrane. However, the functions of insect ankyrin are poorly understood. In this study, the Ankyrin genes of Hyphantria cunea were cloned and their expression characteristics were determined in different developmental stages and tissues of H. cunea. The mortality rate of H. cunea was measured under the stress of exposure to nucleopolyhedrosis virus, which provided a theoretical basis for further developing synergists for NPV-based control strategies. 【Method】 The Ankyrin1 (HcAnk1) and Ankyrin2 (HcAnk2) genes were screened by a transcriptome library, and the characteristics of the two Ankyrin genes were determined using bioinformatics. Using qRT-PCR, the expression of HcAnk1 and HcAnk2 genes were determined at different developmental stages and in different tissues, and under different HcNPV concentrations. The survival rate of H. cunea larvae under HcNPV stress was investigated after the gene silencing of HcAnk1 and HcAnk2 using an RNAi technique. 【Result】 The open reading frames of the HcAnk1 and HcAnk2 genes were 1392 and 1866 bp, encoding 463 and 621 amino acids, respectively. The molecular weights of the HcAnk1 and HcAnk2 proteins were 59.18 and 69.19 ku, respectively, and the theoretical isoelectric points were 5.74 and 8.66, respectively. A phylogenetic analysis showed that HcAnk1 was closely related to Trichoplusia ni and was clustered into one group. HcAnk1 and HcAnk2 were expressed at all developmental stages, with the highest expression in the egg stage and the lowest expression in the sixth instar larva. The highest expression of HcAnk1 and HcAnk2 was observed in the midgut and hindgut, respectively, but HcAnk1 was not detected in the ovary or testis, while HcAnk2 was mainly expressed in the midgut and testis. In tests with different levels of HcNPV stress, the transcription levels of HcAnk1 and HcAnk2 were induced at low concentrations and inhibited at high concentrations. The relative growth rate and food conversion efficiency (ECD) were significantly decreased after silencing HcAnks in H. cunea larvae. Additionally, the H. cunea with HcAnks silencing were significantly less resistant to HcNPV. 【Conclusion】 HcAnk1 and HcAnk2 play an important role in the resistance to HcNPV. HcAnk1 and HcAnk2 can be used as HcNPV synergists for the pollution-free control of H. cunea.

Key words: Hyphantria cunea, Ankyrins, Hyphantria cunea nucleopolyhedrovirus, RNAi

CLC Number:

S763.3

PENG Mengmeng, WU Hongqu, ZHANG Jiawen, YAN Liqiong, CAO Chuanwang, SUN Lili. HcAnk1 and HcAnk2 genes function and HcNPV susceptibility of Hyphantria cunea based on RNAi[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2024, 48(3): 181-190.

Figures/Tables 7

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gene 基因	forward primer sequences 正向引物序列	reverse primer sequences 反向引物序列	primer usage 引物用途
dsHcAnk1	TAATACGACTCACTATAGG GATGCCTTCAGAAGTGATATCAG	TAATACGACTCACTATAGG GAAAAGTGAGCAGCAACTCCA	RNAi
dsHcAnk2	TAATACGACTCACTATAGGG CAGAAAACTATGACATGATTACAT	TAATACGACTCACTATAGGG CAACATCGCCTATTTCTTCG
dsGFP	TAATACGACTCACTATAGGG GGAGAAGAACTTTTCACTGG	TAATACGACTCACTATAGGG AGTTGAACGGATCCATCTTC	RT-qPCR
HcAnk1	CGTGGCTCCTAGCGCTGAGG	GGAGTCTTAGCACGGTGCATTAG
HcAnk2	ATGAATCCATCCTTGGAAGACA	GCAGTGGTTTTGTTTAGAAGATC

treatment 处理	relative growth ratio (RGR) 相对生长率	relative consumption rate (RCR) 相对取食量	efficiency of ingested food (ECI) 食物利用率	efficiency of conversion of food (ECD) 食物转化率
dsGFP	74.40±0.62 a	5.92±0.71 a	12.78±1.73 a	13.56±1.84 a
dsHcAnk1	48.33±0.68 c	4.80±0.13 b	10.09±0.22 a	10.40±0.34 b
dsHcAnk2	60.14±2.53 b	7.84±1.06 a	7.81±1.09 b	8.19±0.95c

HcAnk1 and HcAnk2 genes function and HcNPV susceptibility of Hyphantria cunea based on RNAi

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