Crops ›› 2023, Vol. 39 ›› Issue (2): 46-50.doi: 10.16035/j.issn.1001-7283.2023.02.007

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Optimization of Efficient Silencing System of Tomato VIGS Based on PDS Gene

Guo Yan1(), Liu Zhida1(), Kang Liru2, Bao Tingting1, Yang Xu1, Zhao Jun1, Zhang Zhiwei1()   

  1. 1College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia, China
    2Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010031, Inner Mongolia, China
  • Received:2021-11-16 Revised:2022-01-03 Online:2023-04-15 Published:2023-04-11

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Abstract:

Virus-induced gene silencing (VIGS) is widely used for gene function verification due to its unique advantages. The establishment and optimization of an efficient gene silencing system is the premise and basis for the study of target gene function. In this study, the effects of different OD values, different infection methods, and different culture temperatures after being infected of Agrobacterium on the silencing rate of the PDS gene were studied by using the money maker cultivar and the TRV virus vector carrying phytoene desaturase (PDS) gene sequence. The results showed that, when the OD values of Agrobacterium were 0.50, 1.00, and 1.50, the silencing rates of the PDS gene were 5%, 13%, and 6%, respectively. When the OD value was 1.00, the silencing effects was compared by injection method, vacuum infiltration method and vacuum infiltration after 24 hours of injection treatment, the results showed that, the rates of PDS gene silencing were 13%, 30%, and 15%, respectively. The culture temperature after infection also had a significant effects on the gene silencing rate, with 13% of the plants at 25°C and 46% of the silencing plants at 22°C. The above studies showed that, when the OD value of Agrobacterium is 1.00, the vacuum infiltration was carried out after injection for 24 hours, and the growth of plants at 22°C after inoculation could improve the silencing rate of tomato target genes.

Key words: Tomato, Virus-induced gene silencing, Phytoene desaturase gene, Silence strain rate, Optimization

Fig.1

Phenotype of plants silenced by PDS gene"

Fig.2

Analysis of PDS gene expression in photobleached plants The different lowercase letters mean significant difference (P < 0.05), the same below"

Fig.3

Effects of A.tumefaciens concentration on PDS gene silence strain rate"

Fig.4

Effects of A.tumefaciens different inoculation methods on silence strain rate"

Fig.5

Effects of different temperature treatments on silence strain rate"

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