云南柑橘5种常见病害发生分布调查及其规律分析

doi:10.16420/j.issn.0513-353x.2023-0766

摘要/Abstract

摘要：

为明确黄龙病菌（“Candidatus Liberibacter asiaticus”，CLas）、柑橘黄化脉明病毒（citrus yellow vein clearing virus，CYVCV）、柑橘碎叶病毒（citrus tatter leaf virus，CTLV）、柑橘裂皮类病毒（citrus exocortis viroid，CEVd）和柑橘衰退病病毒（citrus tristeza virus，CTV）所引起的柑橘病害在云南省的发生和分布情况，广泛收集了该省柑橘14个主产州、市38个县的577份柑橘叶片样品，采用巢式PCR和RT-PCR对以上5种病原进行检测，并通过遗传进化分析构建系统发育树。结果表明，CLas、CYVCV、CTLV、CEVd、CTV检出率分别为36.0%、33.6%、15.1%、13.0%、82.0%，其中CLas和CTV在14个供检州、市中均被检测到。从种类上看，CYVCV、CLas和CTV的检出率均为甜橙 > 宽皮柑橘 > 柠檬，其中CTV在柠檬中的检出率（40.0%）明显低于宽皮柑橘（87.4%）和甜橙（92.2%），而CTLV和CEVd在甜橙中的检出率最低，分别为5.9%和4.0%；从海拔上看，CLas的发生与海拔呈现出一定的负相关。此外，不同病害之间存在一定的相关性，CYVCV与CTV、CTLV与CEVd经常存在复合感染的情况，而CLas与CEVd同时侵染的概率较低。通过遗传进化分析发现，云南柑橘病害病原的异地引入和内部传播均为病原扩散的重要途径。

关键词: 柑橘, 黄龙病, 病毒病, 检测, 分布, 传播

Abstract:

To clarify the occurrence and distribution of citrus diseases caused by“Candidatus Liberibacter asiaticus”（CLas），citrus yellow vein clearing virus（CYVCV），citrus tatter leaf virus （CTLV），citrus exocortis viroid（CEVd）and citrus tristeza virus（CTV）in Yunnan Province. In this study，577 citrus leaf samples were collected from 38 counties of 14 major citrus producing states in Yunnan Province. The pathogens of the above five citrus diseases were detected by nested PCR and RT-PCR，phylogenetic tree was constructed by genetic evolution analysis. The analysis showed that the positive rates of CLas，CYVCV，CTLV，CEVd and CTV were 36.0%，33.6%，15.1%，13.0% and 82.0%，respectively. CLas and CTV occurred in all the detected cities. From the citrus species，sweet orange exhibited the highest detection rates of CYVCV，CLas and CTV，followed by mandarin and lemon. Notably，the detection rate of CTV in lemon（40.0%）was obviously lower than that in mandarin（87.4%）and sweet orange（92.2%）. While the detection rate of CTLV and CEVd in sweet orange was only 5.9% and 4.0%，respectively，and was lower than mandarin and lemon. Based on the correlation analysis，results revealed a negative correlation between CLas occurence and altitude. It was also found that the correlations of CYVCV and CTV，CTLV and CEVd occurs frequently，whereas the correlation of CLas and CEVd was rarely detected，suggesting correlations among citrus diseases. Through genetic evolution analysis，it was found that the introduction and internal spread of pathogens in Yunnan were both important ways for the spread of the pathogens.

Key words: citrus, Huanglong disease, virus disease, detection, distribution, spread

聂睿涵, 孙曼丽, 冯迪, 宋谢天, 高俊燕, 郭莉娜, 邓秀新, 柴利军, 谢宗周, 叶俊丽. 云南柑橘5种常见病害发生分布调查及其规律分析[J]. 园艺学报, 2024, 51(11): 2685-2700.

NIE Ruihan, SUN Manli, FENG Di, SONG Xietian, GAO Junyan, GUO Lina, DENG Xiuxin, CHAI Lijun, XIE Zongzhou, YE Junli. An Investigation on Occurrence and Distribution Patterns of Five Common Citrus Diseases in Yunnan[J]. Acta Horticulturae Sinica, 2024, 51(11): 2685-2700.

图/表 10

参考文献 40

[1]	Bai Bin. 2014. Huanglong disease caused 10 000 mu citrus damage in Binchuan,Yunnan. China Fruit Industry Information, 31 (3):67. (in Chinese)
	柏斌. 2014. 云南·黄龙病导致宾川万亩柑桔受害. 中国果业信息, 31 (3):67.
[2]	Chen H, Li Z, Wang X, Zhou Y, Tang K, Zhou C, Zhao X, Yue J. 2014. First report of citrus yellow vein clearing virus on lemon in Yunnan,China. Plant Disease, 98 (12):1747.
[3]	Chen Y, Yi L, Zhong K, Wang C, Chen B, Li S. 2023. Population genetic characteristics of citrus tristeza virus from wild mandarins in the Nanling Mountains of China. Tropical Plant Pathology, 48:270-282.
[4]	Cui Tiantian, Wang Yanjiao, Li Zhongan, Zhou Changyong, Song Zhen. 2017. Bioinformatics and subcellular localization on Citrus yellow vein clearing virus-triple gene block. Acta Horticulturae Sinica, 44 (8):1579-1588. (in Chinese) doi: 10.16420/j.issn.0513-353x.2017-0165
	崔甜甜, 王艳娇, 李中安, 周常勇, 宋震. 2017. 柑橘黄化脉明病毒的TGB基因生物信息学分析及亚细胞定位. 园艺学报, 44 (8):1579-1588.
[5]	Deng Xiuxin. 2022. A review and perspective for citrus breeding in China during the last six decades. Acta Horticulturae Sinica, 49 (10):2063-2074. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0701
	邓秀新. 2022. 中国柑橘育种60年回顾与展望. 园艺学报, 49 (10):2063-2074. doi: 10.16420/j.issn.0513-353x.2021-0701
[6]	Dong Jiahong, Yue Jianqiang, Yin Yueyan, Gao Junyan, Chen Wei, Zhang Zhongkai. 2008. Detection and analysis of lemon tristeza diseases in Yunnan. Journal of Yunnan University(Natural Science Edition),(S1):73-76,80. (in Chinese)
	董家红, 岳建强, 尹跃艳, 高俊燕, 陈伟, 张仲凯. 2008. 云南柠檬生产区衰退病检测与分析. 云南大学学报(自然科学版),(S1):73-76,80.
[7]	Dong Peng, Bao Gaili, Ran Zhiwei, Chen Hairu, Li Fan. 2011. Detection and sequence analysis of the 16S rDNA of citrus huanglongbing causal agent in Yunnan. Journal of Yunnan Agricultural University Science, 26 (5):607-611. (in Chinese)
	董鹏, 包改丽, 冉志伟, 陈海如, 李凡. 2011. 云南柑橘黄龙病病原检测及其16S rDNA序列分析. 云南农业大学学报(自然科学版), 26 (5):607-611.
[8]	Geng Xuexin, Gao Fanglan, Liang Wenxin, Liang Zhouyong, He Xinhua, Zou Chengwu. 2023. Survey and genetic diversity analysis of citrus tristeza virus(CTV)and citrus yellow vein clearing virus(CYVCV)in Guangxi,China. Plant Protection, 49 (4):84-91. (in Chinese)
	耿雪欣, 高方栏, 梁文馨, 梁洲勇, 何新华, 邹承武. 2023. 广西柑橘衰退病和黄脉病调查及其病原病毒的遗传多样性分析. 植物保护, 49 (4):84-91.
[9]	Gross H, Krupp G, Domdey H, Raba M, Jank P, Lossow C, Alberty H, Ramm K, Sanger H. 1982. Nucleotide sequence and secondary structure of citrus exocortis and chrysanthemum stunt viroid. European Journal of Biochemistry, 121:249/257. pmid: 7060550
[10]	Han Jian. 2008. Damage investigation of CTV-Stem pitting and screening of mild CTV strains in Hunan[M. D. Dissertation]. Changsha: Hunan Agricultural University. (in Chinese)
	韩健. 2008. 湖南省柑橘茎陷点型衰退病为害情况调查及其弱毒系筛选研究[硕士论文]. 长沙: 湖南农业大学.
[11]	Huang Linxiao, Yang Yi, Li Danyang, Ma Yunlong, Li Shuxiang, Jiang Lei. 2022. Comparative study on PCR detection primers of citrus HLB. Plant Quarantine, 36 (2):1-6. (in Chinese)
	黄霖霄, 杨毅, 李丹阳, 马云龙, 李树香, 姜蕾. 2022. 柑橘黄龙病PCR检测引物比较研究. 植物检疫, 36 (2):1-6.
[12]	Investigation Group of Citrus Huanglongbing in Yunnan. 1981. Investigation report on citrus yellowing in Yunnan. Chinese Citrus,(4):14-16. (in Chinese)
	云南柑桔黄龙病调查组. 1981. 云南柑桔黄化问题考察报告. 中国柑桔,(4):14-16.
[13]	Jiang Meiliang, Wu Xingjia, He Yuanwei. 2015. Identification of citrus exocortis viroid and its control and prevention technology. Contemporary Horticulture,(13):118-119. (in Chinese)
	蒋美亮, 伍兴甲, 何源委. 2015. 柑桔裂皮病的识别及其控防技术. 现代园艺,(13):118-119.
[14]	Lei Ping, Huang Binbin, Huang Jun, Bi Shiyu, Guo Zhaohui, Liu Qingshu, Tang Ying. 2020. PCR detection of citrus Huanglongbing pathogen and endogenous bacteria isolation for navel orange(Citrus sinensis Osb. var. brasliliensis Tanaka)in Hunan. Hunan Agricultural Sciences,(7):1-4. (in Chinese)
	雷平, 黄彬彬, 黄军, 毕世宇, 郭照辉, 刘清术, 唐滢. 2020. 湘南脐橙柑橘黄龙病的PCR检测及内生细菌的分离. 湖南农业科学,(7):1-4.
[15]	Li Yan,Li Zhao,Han Weijun. 2020. Occurrence of citrus quarantine diseases and the related control technologies in Yunnan Province. Plant Quarantine, 34 (6):53-55. (in Chinese)
	李燕, 李钊, 韩伟君. 2020. 云南省柑橘检疫性病害发生与防控技术. 植物检疫, 34 (6):53-55.
[16]	Li Zhenxi, Pan Ruixuan, Xu Meirong, Zheng Zheng, Deng Xiaoling. 2023. Development of duplex real-time PCR assay of‘Candidatus Liberibacter asiatics’. Acta Horticulturae Sinica, 50 (1):188-196. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0502
	李镇希, 潘睿翾, 许美容, 郑正, 邓晓玲. 2023. 柑橘黄龙病菌双重实时荧光PCR检测方法的建立. 园艺学报, 50 (1):188-196. doi: 10.16420/j.issn.0513-353x.2021-0502
[17]	Lin Youjian, Xie Lianhui, Powell C. 2001. Advances in citrus tristeza virus transmission by brown citrus aphid. Journal of Fujian Agricultural University,(1):59-66. (in Chinese)
	林尤剑, 谢联辉,Powell C. 2001. 橘蚜传播柑橘衰退病毒的研究进展. 福建农业大学学报,(1):59-66.
[18]	Liu Kehong, Song Zhen, Zhou Yan, Li Zhongan, Zhou Changyong. 2012. Influence of eight citrus cultivars on the composition of citrus tatter leaf virus. Journal of Fruit Tree, 29 (1):90-93. (in Chinese)
	刘科宏, 宋震, 周彦, 李中安, 周常勇. 2012. 8种柑橘类植物对柑橘碎叶病毒分子组成的影响. 果树学报, 29 (1):90-93.
[19]	Ma Zhimin, Xu Jianjian, Duan Yu, Wang Chunqing, Su Yue, Zhang Qi, Bin Yu, Zhou Changyong. 2021. Establishment of RT-RPA for citrus yellow vein clearing virus(CYVCV)detection. Chinese Agricultural Sciences, 54 (15):3241-3249. (in Chinese)
	马志敏, 许建建, 段玉, 王春庆, 苏越, 张琦, 宾羽, 周常勇. 2021. 柑橘黄化脉明病毒RT-RPA检测方法的建立. 中国农业科学, 54 (15):3241-3249. doi: 10.3864/j.issn.0578-1752.2021.15.009
[20]	Magome H, Yoshikawa N, Takahashi T, Ito T, Miyakawa T. 1997. Molecular variability of the genomes of capillo viruses from apple,Japanese pear,European pear,and citrus trees. Phytopathology, 87 (4):389-396. pmid: 18945117
[21]	Osvaldo L, Gian P, Vera M, Addolorata C. 2003. An isolate of apple stem grooving virus associated with cleopatra mandarin fruit intumescence. Fitopatologia Brasileira, 28 (1):54-58.
[22]	Pallas V, Flores R. 1989. Interactions between citrus exocortis and potato spindle tuber viroids in plants of Gynura aurantiaca and Lycopersicon esculentum. Intervirology, 30 (1):10-17.
[23]	Schwarz R. 1966. Mechanical transmission of a virus from Meyer lemon to herbaceous hosts. South African Journal of Agriculture Science, 9:263-264.
[24]	Serra P, Seyed M, Bani H, Fagoaga C, Romero J, Susana R, Maria T, Edson B. 2014. Virus-viroid interactions:Citrus tristeza virus enhances the accumulation of Citrus dwarfing viroid in Mexican lime via virus-encoded silencing suppressors. Journal of Virology, 88 (2):1394-1397.
[25]	Shen Lili, Wang Fenglong, Qian Yumei, Chen Dexin, Wu Huihui, He Jingmei,Niu, Zhufeng. 2007. A study on resistance of antagonistic bacteria to TMV. Chinese Tobacco Science,(5):9-11. (in Chinese)
	申莉莉, 王凤龙, 钱玉梅, 陈德鑫, 吴惠惠, 何京美, 牛柱峰. 2007. 拮抗细菌对烟草花叶病毒(TMV)的抑制作用研究. 中国烟草科学,(5):9-11.
[26]	Song Z, Liu K, Li Z, Zhou C. 2016. RT-PCR-RFLP for genetic diversity analysis of the citrus tatter leaf virus strain of apple stem grooving virus. European Journal of Plant Pathology, 144 (3):687-692.
[27]	Syller J, Marczewski W. 2001. Potato leafroll virus-assisted aphid transmission of potato spindle tuber viroid to potato leafroll virus-resistant potato. Journal of Phytopatholigy, 159 (34):195-201.
[28]	Tang Mingli. 2005. Citrus exocortis viroid and its control. Guangxi Horticulture,(2):33-34. (in Chinese)
	唐明丽. 2005. 柑桔裂皮病及其防治. 广西园艺,(2):33-34.
[29]	Tang Xianshou, He Shizhong. 2015. Correlation analysis of key factors in the outbreak and spread of citrus Huanglong disease. China Tropical Agriculture,(4):54-57. (in Chinese)
	唐仙寿, 何世忠. 2015. 柑桔黄龙病暴发蔓延关键因素的关联性分析. 中国热带农业,(4):54-57.
[30]	Wang Fang, Yin Youping, Sun Liqin, Wu Xiaofang, Wang Zhongkang. 2014. Endophytic bacterial community in the symptoms and symptomless tissues of HLB-affected citrus plant. Journal of Microbiology, 54 (8):868-875. (in Chinese)
	王芳, 殷幼平, 孙丽琴, 吴晓芳, 王中康. 2014. 柑橘黄龙病罹病植株显症差异组织内生细菌群落结构分析. 微生物学报, 54 (8):868-875.
[31]	Wu Fengnian, Chen Yanling, Huang Jiaquan, Deng Xiaoling, Cen Yijing. 2016. Effects of temperature and host on Cacopsylla citrisuga growth and development. Journal of South China Agricultural University, 37 (5):57-61. (in Chinese)
	吴丰年, 陈燕玲, 黄家权, 邓晓玲, 岑伊静. 2016. 温度和寄主对柚喀木虱生长发育的影响. 华南农业大学学报, 37 (5):57-61.
[32]	Yang Binhua, Yan Yin, Hu Zhonggui. 2020. Investigation and analysis of the effect of citrus tristeza virus on the growth and fruiting condition of navel orange. Contemporary Horticulture, 43 (3):38-39. (in Chinese)
	杨斌华, 严颖, 胡重贵. 2020. 柑桔衰退病对脐橙生长和结果情况影响的调查与分析. 现代园艺, 43 (3):38-39.
[33]	Yi Long, Tao Zhenzhen, Lu Zhanjun, Zhong Balian. 2012. Sequence analysis of the full length cDNA of a citrus exocortis viroid CEVd-RJ Jiangxi isolate. Fruit trees of southern China, 41 (6):1-4. (in Chinese)
	易龙, 陶珍珍, 卢占军, 钟八莲. 2012. 柑桔裂皮类病毒江西分离物(CEVd-RJ)全长cDNA序列分析. 中国南方果树, 41 (6):1-4.
[34]	Zhang Shaoran, Zong Qi, Liu Yang, Jiang Ling. 2023. In situ hybridization analysis of distinctive ribonucleotide reductase β subunit gene from Candidatus liberibacter asiaticus of citrus. Acta Horticulturae Sinica, 50 (12):2689-2700. (in Chinese) doi: 10.16420/j.issn.0513-353x.2022-1095
	张少然, 宗琪, 刘阳, 姜玲. 2023. 柑橘黄龙病病原菌nrdB基因的原位杂交分析. 园艺学报, 50 (12):2689-2700. doi: 10.16420/j.issn.0513-353x.2022-1095
[35]	Zhao Hengyan, Guan Guijing, Zhou Changyong, Yu Yunqi, Wang Hongsu, Li Zhongan, Liu Jinxiang. 2017. Establishment and application of duplex RT-PCR for the detection of citrus yellow vein clearing virus and citrus tristeza virus. Acta Horticulturae Sinica, 44 (7):1405-1414. (in Chinese) doi: 10.16420/j.issn.0513-353x.2017-0101
	赵恒燕, 关桂静, 周常勇, 于云奇, 王洪苏, 李中安, 刘金香. 2017. 柑橘黄化脉明病毒和衰退病毒的二重RT-PCR检测体系的建立与应用. 园艺学报, 44 (7):1405-1414. doi: 10.16420/j.issn.0513-353x.2017-0101
[36]	Zhao Jun, Guo Jun,Lai Xinpu,Wang Ziran,Mao Jiamei,Yue Jianqiang. 2022. A survey of the occurrence of Huanglongbing and its insect vectors in lemon orchards of western Yunnan. Tropical Agricultural Sciences, 42 (1):61-64. (in Chinese)
	赵俊, 郭俊, 赖新朴, 王自然, 毛加梅, 岳建强. 2022. 滇西柠檬园黄龙病及其媒介昆虫的发生情况调查. 热带农业科学, 42 (1):61-64.
[37]	Zhao Jun,Liu Yueming,Feng Zhijin,Li Yongping,Li Ming,Li Yueqiong. 2018. Analysis of competitive advantages of citrus industry development in Yunnan Province. Yunnan Agriculture,(1):40-41. (in Chinese)
	赵俊, 刘跃明, 冯稚进, 李永平, 李明, 李跃琼. 2018. 云南省柑橘产业发展竞争优势分析. 云南农业,(1):40-41.
[38]	Zhao Xiaolong, Zhang Shenan, Liu Shengqiu, Yang Yanmi. 2005. Citrus tatter leaf virus and its control. Guangxi Horticulture,(2):32-33. (in Chinese)
	赵小龙, 张社南, 刘升球, 阳廷密. 2005. 柑桔碎叶病及其防治. 广西园艺,(2):32-33.
[39]	Zheng Lin,Wang Shuai,Liu Yunuo,Du Meixia,Peng Aihong,He Yongrui,Chen Shanchun,Zou Xiuping. 2022. Gene cloning and expression analysis of NAC gene in citrus in response to Huanglongbing. Acta Horticulturae Sinica, 49 (7):1441-1457. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0553
	郑林, 王帅, 刘语诺, 杜美霞, 彭爱红, 何永睿, 陈善春, 邹修平. 2022. 柑橘响应黄龙病菌侵染的NAC基因的克隆及表达分析. 园艺学报, 49 (7):1441-1457. doi: 10.16420/j.issn.0513-353x.2021-0553
[40]	Zhou Dongguo, Dong Meichao, Li Jinxue, Du Yuxia, Li Jing, Zhu Chunhua, Yue Jianqiang. 2014. Effect of different grafted lemon scions on plant growth. Jiangsu Agricultural Sciences, 42 (7):151-154. (in Chinese)
	周东果, 董美超, 李进学, 杜玉霞, 李晶, 朱春华, 岳建强. 2014. 嫁接不同柠檬接穗对植株生长的影响. 江苏农业科学, 42 (7):151-154.

采样地点 Sampling location		样品 Sample		样品数 Number of samples	海拔/m Altitude	经纬度/° Latitude and longitude	引种地 Place of introduction
州、市 City	县 County	样品 Sample		样品数 Number of samples	海拔/m Altitude	经纬度/° Latitude and longitude	引种地 Place of introduction
保山 Baoshan	隆阳Longyang	沃柑，尤力克柠檬 Orah Mandarin，Eureka Lemon		15	608 ~ 690	98.84 ~ 98.88E，24.97 ~ 25.39N	—
保山 Baoshan	昌宁 Channing	沃柑，椪柑 Orah Mandarin，Ponkan		15	797 ~ 1 267	99.36 ~ 99.45E，24.47 ~ 24.96N	—
楚雄 Chuxiong	永仁 Yongren	沃柑，椪柑，晚熟椪柑 Orah Mandarin，Ponkan，Late-maturing Ponkan		15	—	—	—
楚雄 Chuxiong	双柏 Shuangbai	晚熟绿柑，冰糖橙 Late-maturing Green Mandarin，Bingtangcheng		15	689 ~ 1 028	101.56 ~ 101.78E，24.23 ~ 24.29N	—
大理 Dali	宾川 Binchuan	沃柑，岩溪晚芦 Orah Mandarin，Yanxi Luwan		15	1 381 ~ 1 690	100.45 ~ 100.55E，25.72 ~ 25.98N	四川，云南Sichuan，Yunnan
	巍山 Weishan	尤力克柠檬，香水柠檬 Eureka Lemon，Perfume Lemon		15	1 316 ~ 1 689	99.96 ~ 100.28E，25.00 ~ 25.28N	四川，广东，云南 Sichuan，Guangdong，Yunnan
	鹤庆 Heqing	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 274 ~ 1 316	100.41 ~ 100.44E，26.20 ~ 26.26N	重庆，云南 Chongqing，Yunnan
	弥渡 Midu	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 627 ~ 1 701	100.46 ~ 100.58E，25.22 ~ 25.36N	四川，云南 Sichuan，Yunnan
	永平 Yongping	沃柑，柳叶橘，茂谷柑，尤力克柠檬 Orah Mandarin，Liuyeju Tangerine，Murcott，Eureka Lemon		15	1 314 ~ 1 404	99.35 ~ 99.38E，25.31 ~ 25.32N	云南 Yunnan
德宏 Dehong	芒市 Mangshi	香水柠檬 Perfume Lemon		15	110 ~ 930	98.02 ~ 98.99E，24.22 ~ 24.83N	—
德宏 Dehong	梁河 Lianghe	沃柑 Orah Mandarin		17	860 ~ 1 550	98.28 ~ 98.30E，24.80 ~ 24.83N	—
迪庆 Diqing	德钦 Deqin	皱皮柑 Zhoupigan Mandarin		5	1 627	99.35E，28.20N	—
红河 Honghe	建水 Jianshui	砂糖橘，沃柑，椪柑，芦柑，温州蜜柑 Shatangju，Orah Mandarin，Ponkan，Lugan，Satsuma Mandarin		25	1 359 ~ 1 517	101.93 ~ 103.02E，23.52 ~ 24.01N	—
昆明 Kunming	禄劝 Luquan	沃柑，纽荷尔脐橙，红橘 Orah Mandarin，Newhall Navel Orange，Red Tangerine		15	1 409 ~ 1 598	102.50 ~ 102.71E，25.50 ~ 26.23N	—
昆明 Kunming	东川 Dongchuan	香水柠檬，芦柑，纽荷尔脐橙 Perfume Lemon，Lugan，NewhallNavel Orange		15	1 041 ~ 1 512	103.15 ~ 103.39E，25.99 ~ 26.60N	中国台湾，广西，云南 Chinese Taiwan，Guangxi，Yunnan
丽江 Lijiang	永胜 Yongsheng	沃柑 Orah Mandarin		30	1 248 ~ 1 550	100.57 ~ 100.66E，26.24 ~ 26.74N	广东，广西，重庆 Guangdong，Guangxi，Chongqing
丽江 Lijiang	华坪 Huaping	沃柑，香橙，麻柑，椪柑，岩溪晚芦，柳叶橙 Orah Mandarin，Xiangcheng，Magan，Ponkan，Yanxi Wanlu，Liuyecheng Orange		15	1 539 ~ 1 682	101.16 ~ 101.27E，26.54 ~ 26.79N	—
临沧 Lincang	耿马 Gengma	—		5	—	—	—
	凤庆 Fengqing	砂糖橘，明日见，红橘，红美人，沃柑，春见，长叶香橙，香橙，椪柑 Shatangju，Asumi，Red Tangerine，Beni Madonna，Orah Mandarin，Harumi，Changye Xiangcheng，Xiangcheng，Ponkan		15	1 216 ~ 1 526	99.65 ~ 100.06E，24.41 ~ 24.94N	云南，广西 Yunnan，Guangxi
	云县 Yunxian	柳叶橘，椪柑，晚熟绿柑，贡柑，沃柑，砂糖橘，红美人 Liuyeju Tangerine，Ponkan，Late-maturing Green Mandarin，Gonggan，Orah Mandarin，Shatangju，Beni Madonna		15	1 098 ~ 1 263	100.10 ~ 100.18E，24.45 ~ 24.46N	云南，广西 Yunnan，Guangxi
普洱 Puer	景谷 Jinggu	沃柑，红心柚 Orah Mandarin，Red Pomelo		15	999 ~ 1 175	100.35 ~ 100.73E，23.41 ~ 23.68N	云南，广西，重庆 Yunnan，Guangxi，Chongqing
	墨江 Mojiang	沃柑 Orah Mandarin		15	750 ~ 1 588	101.43 ~ 101.52E，23.35 ~ 23.36N	—
	思茅 Simao	椪柑，绿柑，沃柑 Ponkan，Green Mandarin，Orah Mandarin		15	1 156 ~ 1 187	101.29E，22.54 ~ 22.59N	云南 Yunnan
曲靖 Qujing	师宗 Shizong	椪柑，云柠一号 Ponkan，Yunning 1 Lemon		15	836 ~ 1 210	104.14 ~ 104.34E，24.47 ~ 24.61N	—
曲靖 Qujing	罗平 Luoping	沃柑，椪柑，鲁布革柠檬 Orah Mandarin，Ponkan，Lubuge Lemon		15	906 ~ 1 250	104.30 ~ 104.49E，24.68 ~ 25.12N	—
文山 Wenshan	西畴 Xichou	千金橘，沃柑，椪柑，芦柑Qianjinju，Orah Mandarin，Ponkan，Lugan		15	1 088 ~ 1 204	104.57 ~ 104.58E，23.26 ~ 23.30N	广西，云南 Guangxi，Yunnan
	砚山 Yanshan	红美人，沃柑，脆蜜金柑Beni Madonna，Orah Mandarin，Cuimi Kumquat		20	1 088 ~ 1 469	103.71 ~ 103.79E，23.69 ~ 23.81N	广西，云南 Guangxi，Yunnan
	文山 Wenshan	贡柑，沃柑 Gonggan，Orah Mandarin		15	1 098 ~ 1 485	103.90 ~ 104.04E，23.10 ~ 23.56N	浙江，重庆 Zhejiang，Chongqing
	麻栗坡 Malipo	砂糖橘，沃柑，三红蜜柚Shatangju，Orah Mandarin，Sanhong Miyou		25	669 ~ 1 019	104.87 ~ 105.20E，23.19 ~ 23.42N	福建，云南，广西 Fujian，Yunnan，Guangxi
	马关 Maguan	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 385 ~ 1 428	104.31 ~ 104.44E，22.98 ~ 23.04N	—
玉溪 Yuxi	新平 Xinping	沃柑，冰糖橙 Orah Mandarin，Bingtangcheng		15	725 ~ 1 400	101.69 ~ 101.85E，23.78 ~ 24.02N	—
	元江 Yuanjiang	—		15	—	—	—
	华宁 Huaning	兴津蜜柑，大浦蜜柑、宫本，金秋砂糖橘，冰糖橙Okitsu Wase Mandarin，Dapu Satsuma Mandarin，Miyamoto Wase，Jinqiu Shatangju，Bingtangcheng		15	1 106 ~ 1 202	103.01 ~ 103.12E，24.05 ~ 24.26N	重庆，云南 Chongqing，Yunnan
	易门 Yimen		大浦蜜柑，沃柑，金秋砂糖橘 Dapu Satsuma Mandarin，Orah Mandarin，Jinqiu Shatangju	15	1 262 ~ 1 525	102.05 ~ 102.18E，24.50 ~ 24.54N	重庆，湖北 Chongqing，Hubei
	澄江 Chengjiang		沃柑，冰糖橙 Orah Mandarin，Bingtangcheng	15	1 507 ~ 1 603	103.02 ~ 103.06E，24.55 ~ 24.73N	—
	峨山 Eshan		世纪红橘，兴津蜜柑，冰糖橙 Shiji Red Tangerine，Okitsu Wase Mandarin，Bingtangcheng	15	1 134 ~ 1 434	102.18 ~ 102.20E，24.12 ~ 24.27N	湖南，云南 Hunan，Yunnan
昭通 Zhaotong	盐津 Yanjin		—	11	—	—	—
昭通 Zhaotong	永善 Yongshan		沃柑，白橘 Orah Mandarin，White Tangerine	4	398 ~ 1 066	103.51 ~ 103.86E，28.00 ~ 28.27N	—

采样地点 Sampling location		样品 Sample		样品数 Number of samples	海拔/m Altitude	经纬度/° Latitude and longitude	引种地 Place of introduction
州、市 City	县 County	样品 Sample		样品数 Number of samples	海拔/m Altitude	经纬度/° Latitude and longitude	引种地 Place of introduction
保山 Baoshan	隆阳Longyang	沃柑，尤力克柠檬 Orah Mandarin，Eureka Lemon		15	608 ~ 690	98.84 ~ 98.88E，24.97 ~ 25.39N	—
保山 Baoshan	昌宁 Channing	沃柑，椪柑 Orah Mandarin，Ponkan		15	797 ~ 1 267	99.36 ~ 99.45E，24.47 ~ 24.96N	—
楚雄 Chuxiong	永仁 Yongren	沃柑，椪柑，晚熟椪柑 Orah Mandarin，Ponkan，Late-maturing Ponkan		15	—	—	—
楚雄 Chuxiong	双柏 Shuangbai	晚熟绿柑，冰糖橙 Late-maturing Green Mandarin，Bingtangcheng		15	689 ~ 1 028	101.56 ~ 101.78E，24.23 ~ 24.29N	—
大理 Dali	宾川 Binchuan	沃柑，岩溪晚芦 Orah Mandarin，Yanxi Luwan		15	1 381 ~ 1 690	100.45 ~ 100.55E，25.72 ~ 25.98N	四川，云南Sichuan，Yunnan
	巍山 Weishan	尤力克柠檬，香水柠檬 Eureka Lemon，Perfume Lemon		15	1 316 ~ 1 689	99.96 ~ 100.28E，25.00 ~ 25.28N	四川，广东，云南 Sichuan，Guangdong，Yunnan
	鹤庆 Heqing	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 274 ~ 1 316	100.41 ~ 100.44E，26.20 ~ 26.26N	重庆，云南 Chongqing，Yunnan
	弥渡 Midu	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 627 ~ 1 701	100.46 ~ 100.58E，25.22 ~ 25.36N	四川，云南 Sichuan，Yunnan
	永平 Yongping	沃柑，柳叶橘，茂谷柑，尤力克柠檬 Orah Mandarin，Liuyeju Tangerine，Murcott，Eureka Lemon		15	1 314 ~ 1 404	99.35 ~ 99.38E，25.31 ~ 25.32N	云南 Yunnan
德宏 Dehong	芒市 Mangshi	香水柠檬 Perfume Lemon		15	110 ~ 930	98.02 ~ 98.99E，24.22 ~ 24.83N	—
德宏 Dehong	梁河 Lianghe	沃柑 Orah Mandarin		17	860 ~ 1 550	98.28 ~ 98.30E，24.80 ~ 24.83N	—
迪庆 Diqing	德钦 Deqin	皱皮柑 Zhoupigan Mandarin		5	1 627	99.35E，28.20N	—
红河 Honghe	建水 Jianshui	砂糖橘，沃柑，椪柑，芦柑，温州蜜柑 Shatangju，Orah Mandarin，Ponkan，Lugan，Satsuma Mandarin		25	1 359 ~ 1 517	101.93 ~ 103.02E，23.52 ~ 24.01N	—
昆明 Kunming	禄劝 Luquan	沃柑，纽荷尔脐橙，红橘 Orah Mandarin，Newhall Navel Orange，Red Tangerine		15	1 409 ~ 1 598	102.50 ~ 102.71E，25.50 ~ 26.23N	—
昆明 Kunming	东川 Dongchuan	香水柠檬，芦柑，纽荷尔脐橙 Perfume Lemon，Lugan，NewhallNavel Orange		15	1 041 ~ 1 512	103.15 ~ 103.39E，25.99 ~ 26.60N	中国台湾，广西，云南 Chinese Taiwan，Guangxi，Yunnan
丽江 Lijiang	永胜 Yongsheng	沃柑 Orah Mandarin		30	1 248 ~ 1 550	100.57 ~ 100.66E，26.24 ~ 26.74N	广东，广西，重庆 Guangdong，Guangxi，Chongqing
丽江 Lijiang	华坪 Huaping	沃柑，香橙，麻柑，椪柑，岩溪晚芦，柳叶橙 Orah Mandarin，Xiangcheng，Magan，Ponkan，Yanxi Wanlu，Liuyecheng Orange		15	1 539 ~ 1 682	101.16 ~ 101.27E，26.54 ~ 26.79N	—
临沧 Lincang	耿马 Gengma	—		5	—	—	—
	凤庆 Fengqing	砂糖橘，明日见，红橘，红美人，沃柑，春见，长叶香橙，香橙，椪柑 Shatangju，Asumi，Red Tangerine，Beni Madonna，Orah Mandarin，Harumi，Changye Xiangcheng，Xiangcheng，Ponkan		15	1 216 ~ 1 526	99.65 ~ 100.06E，24.41 ~ 24.94N	云南，广西 Yunnan，Guangxi
	云县 Yunxian	柳叶橘，椪柑，晚熟绿柑，贡柑，沃柑，砂糖橘，红美人 Liuyeju Tangerine，Ponkan，Late-maturing Green Mandarin，Gonggan，Orah Mandarin，Shatangju，Beni Madonna		15	1 098 ~ 1 263	100.10 ~ 100.18E，24.45 ~ 24.46N	云南，广西 Yunnan，Guangxi
普洱 Puer	景谷 Jinggu	沃柑，红心柚 Orah Mandarin，Red Pomelo		15	999 ~ 1 175	100.35 ~ 100.73E，23.41 ~ 23.68N	云南，广西，重庆 Yunnan，Guangxi，Chongqing
	墨江 Mojiang	沃柑 Orah Mandarin		15	750 ~ 1 588	101.43 ~ 101.52E，23.35 ~ 23.36N	—
	思茅 Simao	椪柑，绿柑，沃柑 Ponkan，Green Mandarin，Orah Mandarin		15	1 156 ~ 1 187	101.29E，22.54 ~ 22.59N	云南 Yunnan
曲靖 Qujing	师宗 Shizong	椪柑，云柠一号 Ponkan，Yunning 1 Lemon		15	836 ~ 1 210	104.14 ~ 104.34E，24.47 ~ 24.61N	—
曲靖 Qujing	罗平 Luoping	沃柑，椪柑，鲁布革柠檬 Orah Mandarin，Ponkan，Lubuge Lemon		15	906 ~ 1 250	104.30 ~ 104.49E，24.68 ~ 25.12N	—
文山 Wenshan	西畴 Xichou	千金橘，沃柑，椪柑，芦柑Qianjinju，Orah Mandarin，Ponkan，Lugan		15	1 088 ~ 1 204	104.57 ~ 104.58E，23.26 ~ 23.30N	广西，云南 Guangxi，Yunnan
	砚山 Yanshan	红美人，沃柑，脆蜜金柑Beni Madonna，Orah Mandarin，Cuimi Kumquat		20	1 088 ~ 1 469	103.71 ~ 103.79E，23.69 ~ 23.81N	广西，云南 Guangxi，Yunnan
	文山 Wenshan	贡柑，沃柑 Gonggan，Orah Mandarin		15	1 098 ~ 1 485	103.90 ~ 104.04E，23.10 ~ 23.56N	浙江，重庆 Zhejiang，Chongqing
	麻栗坡 Malipo	砂糖橘，沃柑，三红蜜柚Shatangju，Orah Mandarin，Sanhong Miyou		25	669 ~ 1 019	104.87 ~ 105.20E，23.19 ~ 23.42N	福建，云南，广西 Fujian，Yunnan，Guangxi
	马关 Maguan	沃柑，椪柑 Orah Mandarin，Ponkan		15	1 385 ~ 1 428	104.31 ~ 104.44E，22.98 ~ 23.04N	—
玉溪 Yuxi	新平 Xinping	沃柑，冰糖橙 Orah Mandarin，Bingtangcheng		15	725 ~ 1 400	101.69 ~ 101.85E，23.78 ~ 24.02N	—
	元江 Yuanjiang	—		15	—	—	—
	华宁 Huaning	兴津蜜柑，大浦蜜柑、宫本，金秋砂糖橘，冰糖橙Okitsu Wase Mandarin，Dapu Satsuma Mandarin，Miyamoto Wase，Jinqiu Shatangju，Bingtangcheng		15	1 106 ~ 1 202	103.01 ~ 103.12E，24.05 ~ 24.26N	重庆，云南 Chongqing，Yunnan
	易门 Yimen		大浦蜜柑，沃柑，金秋砂糖橘 Dapu Satsuma Mandarin，Orah Mandarin，Jinqiu Shatangju	15	1 262 ~ 1 525	102.05 ~ 102.18E，24.50 ~ 24.54N	重庆，湖北 Chongqing，Hubei
	澄江 Chengjiang		沃柑，冰糖橙 Orah Mandarin，Bingtangcheng	15	1 507 ~ 1 603	103.02 ~ 103.06E，24.55 ~ 24.73N	—
	峨山 Eshan		世纪红橘，兴津蜜柑，冰糖橙 Shiji Red Tangerine，Okitsu Wase Mandarin，Bingtangcheng	15	1 134 ~ 1 434	102.18 ~ 102.20E，24.12 ~ 24.27N	湖南，云南 Hunan，Yunnan
昭通 Zhaotong	盐津 Yanjin		—	11	—	—	—
昭通 Zhaotong	永善 Yongshan		沃柑，白橘 Orah Mandarin，White Tangerine	4	398 ~ 1 066	103.51 ~ 103.86E，28.00 ~ 28.27N	—

病原 Pathogens	引物序列（5′-3′） Primer	退火温度/℃ Annealing temperature	产物长度/bp The amplified product length	参考文献 Reference
CLas 一轮 First round of CLas	F：GCGCGTATGCAATACGAGCGGCA R：GCCTCGCGACTTCGCAACCCAT	60	1 169	雷平等,2020
CLas 二轮 Second round of CLas	F：RGGGAAAGATTTTATTGGAG R：GAAAATAYCATCTCTGATATCGT	58	750
CYVCV	F：TACCGCAGCTATCCATTTCC R：GCAGAAATCCCGAACCACTA	56	614	马志敏等,2021
CTLV	F：AGAGTGGACAAACTCTAGAC R：CCCTCTCAGCTAGAATTGAA	58	614	Magome et al.,1997；Osvaldo et al.,2003
CEVd	F：CCGGGGATCCCTGAAGGACTT R：GGAAACCTGGAGGAAGTCGAG	58	371	Gross et al.,1982
CTV	F：GGACAAACTTTNNTTTCTGTGAACCTTTC R：GATGAAGTGGTGTTCACGGAGAACTC	56	888	韩健,2008

病原 Pathogens	引物序列（5′-3′） Primer	退火温度/℃ Annealing temperature	产物长度/bp The amplified product length	参考文献 Reference
CLas 一轮 First round of CLas	F：GCGCGTATGCAATACGAGCGGCA R：GCCTCGCGACTTCGCAACCCAT	60	1 169	雷平等,2020
CLas 二轮 Second round of CLas	F：RGGGAAAGATTTTATTGGAG R：GAAAATAYCATCTCTGATATCGT	58	750
CYVCV	F：TACCGCAGCTATCCATTTCC R：GCAGAAATCCCGAACCACTA	56	614	马志敏等,2021
CTLV	F：AGAGTGGACAAACTCTAGAC R：CCCTCTCAGCTAGAATTGAA	58	614	Magome et al.,1997；Osvaldo et al.,2003
CEVd	F：CCGGGGATCCCTGAAGGACTT R：GGAAACCTGGAGGAAGTCGAG	58	371	Gross et al.,1982
CTV	F：GGACAAACTTTNNTTTCTGTGAACCTTTC R：GATGAAGTGGTGTTCACGGAGAACTC	56	888	韩健,2008

品种 Species	样品数量 Number of samples	检出率/% Detection rate
品种 Species	样品数量 Number of samples	CLas	CYVCV	CTLV	CEVd	CTV	CTV强毒 Highly toxic of CTV
沃柑Orah Mandarin	200	37.0	33.0	15.0	20.0	83.0	78.5
椪柑Ponkan	46	34.8	32.6	19.6	10.9	95.6	93.5
晚熟椪柑Late-maturing Ponkan	1	0	100.0	0	0	100.0	100.0
绿柑Green Mandarin	5	20.0	0	40.0	0	100.0	100.0
晚熟绿柑Late-maturing Green Mandarin	6	66.7	83.3	33.3	16.7	100.0	100.0
芦柑Lugan	11	18.2	90.9	0	0	90.9	72.7
岩溪晚芦Yanxi Wanlu	6	0	0	0	0	100.0	100.0
麻柑Magan	1	0	0	0	0	100.0	100.0
茂谷柑Murcott	2	0	0	0	0	50.0	50.0
春见Harumi	1	0	100.0	100.0	0	100.0	100.0
贡柑Gonggan	9	33.3	77.8	11.1	0	100.0	100.0
温州蜜柑Satsuma Mandarin	5	0	40.0	0	0	80.0	60.0
柳叶橘Liuyeju Tangerine	5	20.0	0	40.0	20.0	100.0	100.0
砂糖橘Shatangju	12	58.3	58.3	8.3	0	75.0	66.7
金秋砂糖橘Jinqiu Shatangju	2	50.0	50.0	0	0	100.0	100.0
红橘Red Tangerine	5	60.0	40.0	0	0	80.0	60.0
大浦蜜柑Dapu Satsuma Mandarin	13	38.5	23.1	0	0	100.0	76.9
兴津蜜柑Okitsu Wase Mandarin	10	20.0	10.0	0	0	80.0	60.0
世纪红橘Shiji Red Tangerine	5	20.0	40.0	20.0	0	100.0	100.0
千金橘Qianjinju	7	100.0	28.6	0	0	100.0	28.6
白橘White Tangerine	2	50.0	0	0	0	100.0	100.0
宫本Miyamoto Wase	3	33.3	66.7	0	0	100.0	33.3
明日见Asumi	1	0	100.0	100.0	0	100.0	100.0
红美人Beni Madonna	8	37.5	12.5	75.0	12.5	100.0	100.0
皱皮柑Zhoupigan Mandarin	5	40.0	0	0	0	100.0	100.0
冰糖橙Bingtangcheng	34	47.1	64.7	8.8	2.9	85.3	82.4
柳叶橙Liuyecheng Orange	1	0	0	0	0	100.0	100.0
纽荷尔脐橙Newhall Navel Orange	14	28.6	71.4	0	0	71.4	71.4
香橙Xiangcheng	2	0	50.0	50.0	0	100.0	100.0
长叶香橙Changye Xiangcheng	1	0	100.0	0	0	100.0	100.0
香水柠檬Perfume Lemon	20	0	25.0	0	0	25.0	25.0
尤力克柠檬Eureka Lemon	25	12.0	12.0	20.0	56.0	48.0	20.0
鲁布革柠檬Lubuge Lemon	5	80.0	0	20.0	60.0	100.0	100.0
云柠一号Yuning 1 Lemon	10	50.0	10.0	0	0	10.0	0
红心柚Red Pomelo	5	0	0	40.0	0	100.0	0
三红蜜柚Sanhong Miyou	5	100.0	0	80.0	0	20.0	0
脆蜜金柑Cuimi Kumquat	10	100.0	0	0	0	70.0	70.0
未知品种Unknown species	74	36.5	29.7	20.3	12.2	91.9	63.5
总计Sum	577	36.0	33.6	15.1	13.0	82.0	71.1