Crops ›› 2025, Vol. 41 ›› Issue (6): 45-50.doi: 10.16035/j.issn.1001-7283.2025.06.006

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Construction of a TRV-Mediated VIGS System in Sunflower

Zhou Fei1(), Huang Xutang1, Xie Pengyuan1, Wang Jing1, Liu Yan1, Cui Jiawei2, Tang Lili1, Wang Wenjun1()   

  1. 1 Industrial Crops Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, Heilongjiang, China
    2 Institute of Agricultural Science of the 10th Division, Xinjiang Production and Construction Corps, Beitun 836000, Xinjiang, China
  • Received:2024-06-12 Revised:2024-09-29 Online:2025-12-15 Published:2025-12-12

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Abstract:

To address the limitations of sunflower (Helianthus annuus L.) gene function research due to the lack of a stable genetic transformation system, the sunflower variety ?R5? was used as the material, with the phytoene desaturase (PDS) gene serving as the reporter gene. A recombinant VIGS vector, pTRV2-HaPDS, was constructed and delivered to sunflower cotyledons using two methods: leaf abaxial injection and vacuum infiltration. The results showed that leaf abaxial injection method was unsuccessful. However, the vacuum infiltration method, which involved two cycles of vacuuming to -0.08 MPa for 3 minutes each, was successful. True leaves exhibited partial whitening signs at 20 days after inoculation. By 30 days after inoculation, the albino phenotype became more pronounced, with the majority of plants showing whitening on the second and third pairs of true leaves, though the distribution was uneven. The fourth pair of true leaves showed only a slight albino phenotype. The silencing effect lasted for approximately 20 to 75 days after infection, with a silencing efficiency of about 57%. qRT-PCR results confirmed that the expression of the HaPDS gene was significantly lower in the albino plants compared to the control group, proving successful gene silencing.

Key words: Sunflower, VIGS, HaPDS gene, Gene silencing

Fig.1

The PCR electrophoresis diagram of HaPDS cloning fragments M: DNA marker; 1: HaPDS gene fragment."

Fig.2

Sequence comparison results of pTRV2-HaPDS vector The first line shows reference sequence of HaPDS gene fragment, the second and third line show sequence of positive transformed clone，deep blue indicates the aligned sequences."

Fig.3

PCR identification of pTRV2-HaPDS vector in Agrobacterium 1 and 2: Agrobacterium positive clones; “-”: pure water negative control; “+”: pTRV2-HaPDS vector plasmid positive control; M: DL2000 marker."

Fig.4

Albino phenotype of sunflower leaf after HaPDS gene silencing"

Fig.5

Analysis of HaPDS gene expression in sunflower leaves CK: inoculated water control group; TRV2:Empty: control group of inoculated virus empty vector; TRV2:HaPDS: the experimental group inoculated with HaPDS recombinant virus vector,“*”means significantly different from CK (P < 0.05),“**”means extremely significantly different from CK (P < 0.01)."

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