摘要：

为探究苹果各器官不同部位的RNA提取和病毒检测效果,以携带3种病毒的苹果植株为试验材料,采用吸附柱法提取苹果的花(雌蕊、花药、花丝、花瓣、花萼、花托和花柄)、叶片(叶柄、叶肉和叶脉)、果实(果柄、果皮、果肉、果核、种皮和胚乳)和枝条(韧皮部)4个器官共17个组织部位的总RNA,利用常规RT-PCR对不同部位进行病毒检测,并采用实时荧光定量PCR分析病毒在不同部位的相对含量。结果表明,苹果不同器官的RNA提取效果存在差异,叶片的浓度最高,其次是花和枝条,果实的浓度最低。花药、花丝、花柄在花中提取效果较好、胚乳和叶柄分别在果实和叶片中提取效果最好。除胚乳外,其余16个组织部位均能检测出3种苹果病毒。定量分析表明,苹果病毒在花中的相对含量最高,在雌蕊、种皮和叶柄中的相对含量分别高于花、果实和叶片的其他组织部位。此外,3种病毒在果实和叶片的不同组织部位的含量变化趋势相似,在花的不同组织部位的分布存在一定的差异。

关键词: 苹果, RNA 提取, 病毒检测, 病毒含量比较

Abstract:

In order to explore the effect of RNA extraction and virus detection of different parts of apple organs,apple plants infected by three viruses were used as experimental materials.Adsorption column method was used to extract total RNA from flowers(pistil,anther,filament,petals,calyx,receptacle,and flower stalk),leaves(petiole,mesophyll,and veins),fruits(fruit stalk,peel,flesh,score,episperm,and endosperm)and branches(phloem),a total of 17 tissues of four organs.The extraction effects of different sampling parts were compared,the detection effects of three viruses in different parts of organs were analyzed by regular RT-PCR,and the relative content of viruses in different parts of organs were analyzed by Real-time fluorescence quantitative PCR.The results showed that the effects of RNA extraction from different organs of apple were different,with the highest concentration in leaves,followed by flowers and phloem of branches,and the lowest concentration in fruits.Anther,filament and petiole were best extracted from flowers and endosperm and petiole were best extracted from fruits and leaves,respectively.Three apple viruses could be detected in 16 tissues except endosperm.Quantitative analysis showed that the relative content of the three apple viruses was the highest in flowers.Furthermore,the relative contents of viruses in pistil,episperm and petiole were correspondingly higher than that in other tissue parts of flower,fruit and leaf,respectively.In addition,the content of the three viruses in different tissue parts of fruits and leaves showed similar trends,but there were certain differences in the distribution of different tissue parts of flowers.

Key words: Apple, RNA extraction, Virus detection, Comparison of viral concentration