GTP cyclohydrolase: a RNAi target gene against the brown planthopper (Nilaparvata lugens)

doi:10.3969/j.issn.1004-1524.20240530

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (6): 1285-1292.DOI: 10.3969/j.issn.1004-1524.20240530

• Plant Protection • Previous Articles Next Articles

GTP cyclohydrolase: a RNAi target gene against the brown planthopper (Nilaparvata lugens)

DONG Daixing¹(), YANG Guiying², WANG Aiying², LUO Ju², LIU Shuhua²^,^*()

1. Fuyang District Agricultural Technology Extension Center, Hangzhou City, Hangzhou 311400, China
2. China National Rice Research Institute, Hangzhou 311401, China

Received:2024-06-21 Online:2025-06-25 Published:2025-07-08

Abstract

Abstract:

Guanosine triphosphate cyclohydrolase I (GCHI) is a crucial regulatory factor for the growth and development of holometabolous insects, involved in melanin synthesis and the molting process. To evaluate the feasibility of the GCHI gene as a target for RNA interference (RNAi) in controlling the hemimetabolous insect Nilaparvata lugens(brown planthopper, BPH), this study cloned the NlGCHI gene from BPH, analyzed its spatiotemporal expression patterns, and validated gene function through microinjection experiments. The results indicated that NlGCHI has two transcripts, designated as NlGCHIa and NlGCHIb. Notably, NlGCHIb exhibited periodic expression dynamics in late-instar nymphs and was highly expressed in the cuticle. Treatment of BPH with dsNlGCHI injection or feeding with the GCHI inhibitor 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) resulted in high mortality rates, primarily due to molting defects characterized by failure to shed the old cuticle properly. Additionally, dsNlGCHI-treated individuals displayed abnormal melanization. The above results indicate that the function of GCHI remains relatively conserved in both holometabolous insects and hemimetabolous insect, influencing cuticle formation and molting processes in insects. Its encoding gene can be utilized to develop RNAi-based pesticides for controlling N. lugens.

Key words: Nilaparvata lugens, RNA interference, guanosine triphosphate cyclohydrolase I, cuticle

CLC Number:

S435.112.3

DONG Daixing, YANG Guiying, WANG Aiying, LUO Ju, LIU Shuhua. GTP cyclohydrolase: a RNAi target gene against the brown planthopper (Nilaparvata lugens)[J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1285-1292.

Figures/Tables 5

Table 1 The specific primers used in this research

用途 Purpose	引物名称^a Primer name^a	引物序列(5'→ 3')^b Primer sequence(5'→ 3')^b
ORF扩增	NlGCHI-F1	ATGGAGCTGGACCACAGACCAC
ORF clone	NlGCHI-R1	CAAGAATTCCTCGCGTGTCTTGG
非翻译区扩增RACE	NlGCHI-5'	GTGGTCTGTGGTCCAGCTCCATGTC
	NlGCHI-3'	GCAAGACGACCACATCTACGATGCTG
实时荧光定量PCR	NlGCHIa-F	CACCAGGTCACGAGAAA
qRT-PCR	NlGCHIb-F	CGAGAATGGTCACGAGAA
	NlGCHIa&b-R	AATACGGCGTCATTCAG
	qRPS15-F	CCGATCGTGTGGCGTTGAAGGG
	qRPS15-R	ATGGCCGACATTCTTCCAGGTCC
	qTUB-F	ACTCGTTCGGAGGAGGCACC
	qTUB-R	GTTCCAGGGTGGTGTGGGTGGT
dsRNA合成引物	dsNlGCHI-F	taatacgactcactatagggagGCATCACTTGGTCCCATTC
dsRNA synthesis	dsNlGCHI-R	taatacgactcactatagggagAATTCCTCGCGTGTCTTGG

Fig.1 Spatial-temporal expression patterns of NlGCHIa and NlGCHIb in Nilaparvata lugens (A) Relative expression levels of NlGCHIa and NlGCHIb during different developmental stages. N4-4h represents 4 h-old 4th-instar nymphs; FA-4h represents 4 h-old female adults; The other samples have the same naming method. (B) Relative expression levels of NlGCHIa and NlGCHIb in different tissues. MG, Midgut; FB, Fat body; CT, Cuticle; OV, Ovary; TE, Testis. The relative expression level was expressed as mean ±standard error (n=4).

Fig.2 Effects of dsNlGCHI injection on NlGCHI mRNA expression levels and survival rate in N. lugens (A) Relative expression level of NlGCHI post dsNlGCHI injection; (B) Survival pattern after injection of dsNlGCHI into 3rd instar nymphs. *, The survival rate of BPHs injected with dsNlGCHI is significantly lower than that injected with dsGFP.

Fig.3 Effects of NlGCHI mRNA knockdown on eye coloration, body coloration and dead phenotype in N. lugens (A) The phenotype of compound eyes after dsNlGCHI injection; (B) The phenotype of body color after dsNlGCHI injection; 0 h, 4 h, and 24 h represent 0 h-, 4 h- and 24 h-old 5th-instar nymphs, respectively; (C) The dead phenotype after dsNlGCHI injection.

Fig.4 Effects of inhibitor DAHP on the growth and development of N. lugens (A) The survival rate curve of N. lugens; *, The survival rate of BPHs after feeding inhibitor DAHP is significantly lower than that in the control group; (B) The dead phenotype of N. lugens after feeding the inhibitor DAHP.

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GTP cyclohydrolase: a RNAi target gene against the brown planthopper (Nilaparvata lugens)

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