The Citrus tristeza virus (CTV) uses 3 silencing suppressor genes, p20, p23 and p25, to resist the attacks from its Citrus hosts. Inactivating these genes is therefore obviously a potential defensive option in additio...The Citrus tristeza virus (CTV) uses 3 silencing suppressor genes, p20, p23 and p25, to resist the attacks from its Citrus hosts. Inactivating these genes is therefore obviously a potential defensive option in addition to the current control strat-egies including aphid management and the use of mild strain cross protection. In this study, we cloned partial DNA frag-ments from the three genes, and used them to construct vectors for expressing hairpin RNAs (hpRNAs). To facilitate the formation of hpRNAs, the constructs were introduced in a loop structure. Fol owing transformation of sour orange (Citrus aurantium) with these constructs, 8 p20 hpRNA (hp20) and 1 p25 hpRNA (hp25) expressing lines were obtained. The 7 hp20 transgenic lines were further characterized. Their reactions to CTV were tested fol owing inoculation with CT14A and/or TR-L514, both of which are severe strains. Results showed that 3 lines (hp20-5, hp20-6 and hp20-8) were completely resistant to TR-L514 under greenhouse conditions for no detectable viral load was found in their leaves by PCR. However, they exhibited only partial suppression of TR-L514 under screen house conditions since the virus was detected in their leaves, though 2 months later compared to non-transgenic controls. Further tests showed that hp20-5 was tolerant also to CT14A under screen house conditions. The growth of hp20-5 was much better than others including the controls that were concurrently chal enged with CT14A. These results showed that expressing p20 hpRNA was sufifcient to confer sour orange with CTV resistance/tolerance.展开更多
Viral RNAs have been known to contain N^(6)-methyladenosine(m^(6)A)modifications since the 1970s.The function of these modifications remained unknown until the development of genome-wide methods to map m^(6)A residues...Viral RNAs have been known to contain N^(6)-methyladenosine(m^(6)A)modifications since the 1970s.The function of these modifications remained unknown until the development of genome-wide methods to map m^(6)A residues.Increasing evidence has recently revealed a strong association between m^(6)A modifications and plant viral infection.This highlight introduces advances in the roles of RNA m^(6)A modifications in plant-virus interactions.展开更多
基金supported by the International Science & Technology Cooperation Program of China (2012DFA30610)the National Natural Science Foundation of China (30571291)the Special Fund for Agro-Scientific Research in the Public Interest, China (201203075-07)
文摘The Citrus tristeza virus (CTV) uses 3 silencing suppressor genes, p20, p23 and p25, to resist the attacks from its Citrus hosts. Inactivating these genes is therefore obviously a potential defensive option in addition to the current control strat-egies including aphid management and the use of mild strain cross protection. In this study, we cloned partial DNA frag-ments from the three genes, and used them to construct vectors for expressing hairpin RNAs (hpRNAs). To facilitate the formation of hpRNAs, the constructs were introduced in a loop structure. Fol owing transformation of sour orange (Citrus aurantium) with these constructs, 8 p20 hpRNA (hp20) and 1 p25 hpRNA (hp25) expressing lines were obtained. The 7 hp20 transgenic lines were further characterized. Their reactions to CTV were tested fol owing inoculation with CT14A and/or TR-L514, both of which are severe strains. Results showed that 3 lines (hp20-5, hp20-6 and hp20-8) were completely resistant to TR-L514 under greenhouse conditions for no detectable viral load was found in their leaves by PCR. However, they exhibited only partial suppression of TR-L514 under screen house conditions since the virus was detected in their leaves, though 2 months later compared to non-transgenic controls. Further tests showed that hp20-5 was tolerant also to CT14A under screen house conditions. The growth of hp20-5 was much better than others including the controls that were concurrently chal enged with CT14A. These results showed that expressing p20 hpRNA was sufifcient to confer sour orange with CTV resistance/tolerance.
基金funded by the National Key Research and Development Program of China(2021YFD1400400)to FLthe National Natural Science Foundation of China(32320103010 and 31930089)to FL and XZ.
文摘Viral RNAs have been known to contain N^(6)-methyladenosine(m^(6)A)modifications since the 1970s.The function of these modifications remained unknown until the development of genome-wide methods to map m^(6)A residues.Increasing evidence has recently revealed a strong association between m^(6)A modifications and plant viral infection.This highlight introduces advances in the roles of RNA m^(6)A modifications in plant-virus interactions.