摘要
【目的】探究沃柑感染黄龙病的田间症状表现及各症状与感染黄龙病的相关性,为种植者尽早鉴别黄龙病树和及时做好黄龙病防控工作提供参考依据。【方法】从广西不同地区沃柑果园采集沃柑黄龙病疑似样品,对样品按症状分类并记录,用试剂盒法提取叶脉和果柱基因组DNA,采用黄龙病亚洲种的16S r DNA特异引物f D1/fD2和OI1/OI2进行巢式PCR(Nested-PCR)扩增,扩增结果在1%琼脂糖凝胶电泳上检测,统计不同症状样品的阳性率。【结果】收集的214份黄龙病疑似样品中,不同症状样品均检出黄龙病亚洲种病原菌,检出阳性率表现为典型红鼻子果=斑驳黄化叶>果蒂稍红果>均匀黄化叶>叶脉黄化叶>僵果>缺锌黄化叶>其他黄化叶>正常叶。各症状类型与感染黄龙病均存在相关性,但存在差异,其中,典型红鼻子果和斑驳黄化叶阳性率均为100.0%,与感染黄龙病相关性最大;其次果蒂稍红果阳性率为90.9%,与感染黄龙病相关性很大;夏、秋梢均匀黄化叶阳性率为46.5%,与感染黄龙病相关性大。沃柑感染黄龙病后症状复杂,大多出现两种以上复合症状,结果树最初症状为出现果蒂稍红果,落果较多,未结果树以夏、秋梢黄化居多,根系容易受损;正常叶阳性率达16.7%,说明沃柑感染黄龙病早期不易显症。【结论】沃柑出现典型红鼻子果和斑驳黄化叶,可田间诊断为黄龙病树;出现果蒂稍红果,并严重落果,基本可田间诊断为黄龙病树;夏、秋梢均匀黄化黄龙病检出率高,但确诊还需与其他疑似症状(叶脉黄化叶、僵果、缺锌黄化叶、其他黄化叶和正常叶)一样通过PCR进一步检测。Nested-PCR检测技术灵敏性高,可作为沃柑感染黄龙病的早期鉴定技术。
【Objective】The symptoms of Orah infected with Huanglongbing(HLB)and the correlation between the symptoms with HLB were studied in order to provide reference for growers to identify HLB from Orah as early as possible,and to prevent in time.【Method】The leaf samples of suspected HLB in Orah orchards in different places in Guangxi were collected,then classified and recorded by symptoms.Genomic DNA of leaf vein and fruit column was extracted by kit and Nested-PCR amplification was conducted by special primers of 16 S rDNA gene of Candidatus Liberibacter asiaticus(fD1/fD2 and OI1/OI2).The results of PCR amplification were detected by 1%agarose gel electrophoresis and the positive rate of samples with different symptoms was calculated.【Result】Among the 214 suspected samples of HLB collected,the samples of different symptoms were detected Candidatus Liberibacter asiaticus,and the positive rate presented as typical red nose fruits=mottled yellow leaves>fruit stalkslightly red fruit>uniform yellow leaves>leaves with yellow central vein>stone fruits>zinc deficiency yellow leaves>other yellow leaves>normal leaves.All the symptoms were correlated with HLB,but the correlations were different.Red nose fruits and mottled yellow leaves had the positive rate of100.0%and had maximal correlation with HLB.Fruit stalks lightly red fruit had positive rate of 90.9%,which was highly correlated with HLB.Uniform yellow leaves in summer and autumn had positive rate of 46.5%and had strong correlation with HLB.After infection of HLB,the symptoms were complex,most of trees presented two or more complex symptoms.The initial symptoms of trees with fruits were fruit stalks lightly red fruits with a large number of fruit dropping.Most of the unfruited trees were yellowing in summer and autumn,and their roots were easily damaged.The positive rate of normal leaf was 16.7%,indicating that it was not easy to show symptomsin the early period of infection of HLB.【Conclusion】When there are typical"red nose"fruits and mottled yellow leaves on Orah,it can be diagnosed as HLB.When the fruit stalk appears slightly red,and the fruits drop seriously,it can be basically diagnosed as HLB.The detection rate of yellowing summer and autumn shoots is high,but to be diagnosed,it needs to be detected by PCR with other symptoms(leaves with yellow central vein,stone fruits,zinc deficiency yellow leaves,other yellow leaves andnormal leaves.Nested-PCR is a method with high sensitivity,which can be an early identification for Orah infected with HLB.
作者
刘福平
王茜
陈东奎
陈香玲
廖惠红
汪妮娜
黄宏明
黄其椿
LIU Fu-ping;WANG Xi;CHEN Dong-kui;CHEN Xiang-ling;LIAO Hui-hong;WANG Ni-na;HUANG Hong-ming;HUANG Qi-chun(Horticulture Research Institute,Guangxi Academy of Agricultural Sciences/Nanning Investigation Station of South Subtropical Fruit Trees,Ministry of Agriculture and Rural Affairs/Guangxi Citrus Huanglongbing Prevention and Control Engineering Technology Research Center,Nanning 530007,China;Guangxi Crop Genetic Improvement and Biotechnology Laboratory,Nanning 530007,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2020年第1期101-107,共7页
Journal of Southern Agriculture
基金
广西科技重大专项(桂科AA18118046-2，桂科AA17204097-8)
南宁市科学研究与技术开发计划项目(20182198-2)
广西农业科学院基本科研业务专项(桂农科2015YT51)~~
