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柿树为我国重要的经济林果树,柿果、柿叶和柿霜等包含丰富的生物活性成分,具有重要的医学价值。从山东省邹城市采集到表现带化并伴随节间缩短和叶片变小的柿树枝条样品,利用透射电子显微镜进行植原体细胞检测,结果显示染病的柿枝条韧皮部筛管细胞内存在直径为200~600 nm的圆形或者椭圆形植原体粒子。提取染病柿树枝条和健康枝条的韧皮部总DNA,利用植原体16S rRNA基因及延伸因子tuf基因通用引物进行PCR扩增,从染病组织DNA中得到了长度分别约为1.2和0.8 kb的片段,健康柿树枝条没有扩增条带。序列一致率分析表明,柿树带化植原体16S rRNA基因和tuf基因与枣疯植原体相应基因的核苷酸一致率分别高达99.8%和99.9%;i Phy Classifi er在线分析表明,柿树带化植原体与16Sr V-B亚组代表株系的16S rRNA基因具有相同的酶切图谱,相似系数为1.0;基于两个基因构建的系统进化树均表明,柿树带化植原体与16Sr V-B亚组成员聚为一簇。生物学和分子生物学鉴定结果表明,柿树带化病存在植原体,且该植原体属于榆树黄化组(16Sr V)B亚组,与候选种‘Candidatus Phytoplasma ziziphi’相关。
Persimmon tree is an important economic forest tree in our country, persimmon fruit, persimmon leaf and persimmon cream contain rich bioactive ingredients, which has important medical value. From Zoucheng, Shandong Province, samples of persimmon shoots with banding and shortening of internodes and leaves were collected and detected by transmission electron microscopy. The results showed that there were 200 ~ 600 nm round or oval phytoplasma particles. Total phloem DNA extracted from infected persimmon shoots and healthy shoots was amplified by PCR using phytoplasma 16S rRNA gene and elongation factor tuf gene universal primers, and fragments of about 1.2 and 0.8 kb in length were obtained from the infected tissues, respectively, and healthy Persimmon branches without amplification bands. Sequence analysis showed that the nucleotide identity of 16S rRNA, tuf and Phytoplasma counterparts was 99.8% and 99.9%, respectively. On-line analysis by i Phy Classifi er showed that persimmon tree The phylogenetic tree of 16S rRNA gene of the 16Sr VB subgroup was the same as that of the 16S rRNA gene of the 16Sr VB subgroup. The similarity coefficient was 1.0. Phylogenetic tree based on the two genes showed that the phylogenetic tree of 16S rBV was Members clustered together. The results of biology and molecular biology showed that phytoplankton existed in the persimmon belt disease, and the phytoplasma belonged to the elite tree group (16Sr V) B subgroup, which was related to the candidate species Candidatus Phytoplasma ziziphi.