柑橘裂皮病类病毒的一步RT-PCR法检测及其在甜橙体内的分布

Preliminary study on One-step RT-PCR detection of Citrus exocortis viroid (CEVd) and its distribution in different parts of sweet orange

柑橘裂皮病是由柑橘裂皮病类病毒(Citrusexocortisviroid,CEVd)引起的一种重要的柑橘病害。分别采用快速微量核酸提取法和SDS-KAc抽提法在表现症状的Etrog香橼中提取核酸,采用一步RT-PCR法对CEVd进行检测,结果显示SDS-KAc法可以消除带毒样品中非特异性条带。从嫁接接毒的铜水72-1锦橙植株的接穗部取嫩叶、老叶、皮,从砧木部茎杆上取老皮以及根皮分别提取总核酸进行RT-PCR检测,分析CEVd在甜橙体内的分布情况。初步检测结果表明在接穗的嫩叶、老叶、皮,砧木部茎杆的老皮和根皮上都可以检测到CEVd,以接穗部叶和皮检出稳定性高。

Citrus exocortis is an important viroid disease. Preliminary study on comparison of Fast Micro-extraction method and SDS-KAc method,detecting CEVd in Etrog citron with symptoms by One-step RT-PCR, revealed that SDS-KAc method was preferable for eliminating non-specific band. Distribution of CEVd in the young leaf, mature leaf, bark and rootlet was monitored by One-step RT-PCR. The results indicated that CEVd could be detected in all parts monitored by One-step PT-PCR. Considering stability, leaf and bark of scion are more suitable to be used for detection of CEVd by Onestep RT-PCR.