选用分别含有抗番茄黄化曲叶病(TYLCD)基因Ty-1、Ty-2和Ty-3的3种抗病番茄材料(分别简称TY-1、TY-2和TY-3)和感病番茄品种(苏粉9号),利用携带TYLCV的烟粉虱对所选番茄材料幼苗接种病毒,从核酸水平检测抗病品系和感病品种的病毒侵入时间差异,并检测接种病毒前后己糖转运基因(LeHT1)和透性蛋白基因(Permease)在转录水平上的表达量变化。结果显示,感病品种与抗病品系TY-2在接种第2 d便检测到病毒,抗病品系TY-1和TY-3分别在3 d、15 d检测到病毒,说明病毒侵入时间早晚与番茄的抗病性没有直接关系。感病品种和抗病品系TY-1分别在接种后7 d、20 d有明显的发病症状,抗病品系TY-2和TY-3整个生育期均无发病症状。感病品种接种前LeHT1无明表达,接种后随时间增加表达量先上升后下降;抗病品系TY-2、TY-3在接种前有较高的表达量,接种后随时间增加呈下降趋势,抗病品系TY-1在接种后5 d时降至最低,随后稍有增加。接种后,Per-mease基因在感病品种中表达量显著提高,7 d后略有降低;抗病品系TY-1的Permease基因表达量先降低,15 d时表达量回升;抗病品系TY-2的Permease基因在接种后的表达量增加;抗病品系在TY-3中,Permease基因在接种前后表达水平变化不大,直至15 d时略有升高。接种前、后LeHT1和Permease基因在不同抗病材料和感病材料中具有不同的表达特征,表明不同抗源可能含有不同的抗病信号通路和抗病机制。
In order to identify the difference in tomato yellow leaf curl virus(TYLCV) infection time in three resistant tomato lines(R) which contain Ty-1,Ty-2 and Ty-3 gene respectively(hereinafter referred as materials TY-1,TY-2,TY-3) and susceptible tomato line(S) Sufen 9,and to compare the changes of resistance-related gene [hexose transporter gene(LeHT1) and permease 1-like protein gene(permease) ]expression before and after TYLCV infection in R and S lines,PCR and semi-quantitative PCR were used to detect TYLCV infection time and LeHT1 and permease genes expression profile.The results showed that the virus was detectable in S and TY-2 seedling just 2 d after inoculation,and in TY-1 and TY-3 seedling at 3 d and 15 d after inoculation respectively,which indicated that there was no direct relation between the time for virus to invade and disease resistance.S and TY-1 showed typical disease symptoms 7 d and 20 d after inoculation respectively,while TY-2 and TY-3 were symptomless at all time.There was no LeHT1 gene expressed before inoculation in S lines,but the expression increased and then decreased after TYLCV inoculation.LeHT1 gene had a higher expression before inoculation in TY-2 and TY-3,and the expression presented downtrend after TYLCV inoculation.However LeHT1 decreased to minimum at 5 d after inoculation in TY-1,then increased slightly.The expression of permease gene had a significant increase in S,and a slight decrease at 7 d after inoculation.The expression of permease gene decreased after inoculation in TY-1,and the expression went up at 15 d after inoculation.On the contrary,the expression of permease increased in TY-2 after inoculation.The expression of permease gene has a little change before and after inoculation in TY-3,only increased at 15 d after inoculation.The expression pattern of LeHT1 and permease gene was different among these lines before and after inoculation.These indicate that they maybe contain divergent resistant signal pathway and the resistant mechanism.