| 摘要: |
| 【 目的】明确引起茶梅（Camellia sasanqua）花叶病的病毒种类和主要病毒的全基因组序列，建立可靠的病毒分子检测技术，为茶梅病毒病的早期诊断和抗病毒育种提供参考。【 方法】以2022年7月至2024年3月，采自陕西、浙江、福建和江苏地区具有黄化、褪绿、植株矮小等症状的茶梅叶片为材料，通过小RNA（sRNA）测序技术鉴定茶梅花叶病的病毒种类，并克隆病毒的全基因组序列，研究其分子变异情况。建立茶梅3种病毒一步法多重RT-PCR（mRT-PCR）快速诊断技术并进行优化。【 结果】在表现花叶症状的茶梅叶片中检测到山茶环斑相关病毒 3（camellia ringspot-associated virus 3，CRSaV-3）、山茶黄化环斑病毒（camellia yellow ringspot virus，CaYRSV）和山茶乙型线形相关病毒 3（camellia japonica associated metaflexid virus 3，CjBV-3）3种病毒。系统发育结果显示，茶梅3种病毒分子变异较小，但与来源于山茶花（C. japonica）的分离物相比，仍然存在较大的分子变异。建立了3种茶梅病毒一步法 mRTPCR检测技术，该检测技术具有快速、高效、操作简便等优点。【 结论】首次鉴定了茶梅花叶病的病毒种类，克隆获得3种茶梅病毒的11条全基因组序列，建立了茶梅病毒mRT-PCR检测技术，该技术具有快速、高效、操作简便等特点，可用于CRSaV-3、CaYRSV和CjBV-3的快速检测。 |
| 关键词: 茶梅花叶病 病毒鉴定 多重RT-PCR（mRT-PCR） |
| DOI：10.13207/j.jnwafu.2026.04.016 |
| 分类号: |
| 基金项目:国家自然科学基金项目（32000123）；陕西省重点研发计划项目（2024NC-YBXM-058）；陕西省高校青年创新团队项目 （2023AYTD01）；陕 西省教育厅协同创新中心项目（23JY003）；国家级大学生创新训练项目（202211397007， 202411397014） |
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| Identification of virus species of Camellia sasanqua mosaic disease and establishment of multiplex PCR detection technology |
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ZHU Hongpei1,2, HUANG Wenrou1, LIU Huan11, LIU Fei3
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1.College of Modern Agriculture and Biotechnology，Ankang University，Ankang，Shaanxi 725000，China;2.School of Horticulture，Anhui Agricultural University，Hefei，Anhui 230036，China;3.Xianyang Agricultural Research Academy，Xianyang，Shaanxi 712000，China
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| Abstract: |
| 【 Objective】This research aims to determine the species of viruses that cause Camellia sasanqua mosaic disease and the complete genome sequences of the main viruses，and develop reliable molecular detection protocols to facilitate early disease diagnosis and support antiviral breeding.【Method】C. sasanqua leaves were collected from symptomatic plants exhibiting yellowing，chlorosis，and dwarf in Shaanxi，Zhejiang，Fujian，and Jiangsu from July 2022 to March 2024. Small RNA（sRNA） sequencing was used to identify the virus species，and the complete genome sequence was obtained to study its molecular variation. A one-step multiplex RT-PCR（mRTPCR）rapid diagnosis technology for three viruses of C. sasanqua mosaic disease was established and optimized.【Result】Three viruses，including camellia ringspot-associated virus 3（CRSaV-3），camellia yellow ringspot virus （CaYRSV） and camellia japonica associated metaflexid virus 3（ CjBV-3），were identified in C. sasanqua with mosaic symptoms. The phylogenetic analysis revealed that these three viruses in C. sasanqua exhibited relatively low levels of molecular variation，but significant difference still can be observed compared to the isolates derived from C. japonica.In addition，a one-step mRT-PCR detection technique for these three viruses of C. sasanqua mosaic disease was established with advantages of rapidity，high efficiency and simple operation.【Conclusion】This work firstly identified the viruses associated with C. sasanqua mosaic disease，and 11 complete genome sequences of them were cloned successfully. A mRT-PCR detection technology for these virus was established，which was fast，efficient，and easy to operate for detection of CRSaV-3，CaYRSV，and CjBV-3. |
| Key words: Camellia sasanqua mosaic disease virus identification multiplex RT-PCR（mRT-PCR） |
