文献类型：期刊文献

中文题名：嫁接于3个枣园病砧的抗病接穗枣疯植原体检测及分子变异

英文题名：Detection and Molecular Variation of Jujube Witches’Broom Phytoplasma in Resistant Jujube Scions Grafted on Diseased Root Stocks in Three Jujube Orchards

作者：张文鑫[1] 于少帅[2] 田国忠[1] 王合[3] 任争光[4] 王圣洁[1] 孔德治[1] 李永[1] 林彩丽[1]

第一作者：张文鑫

机构：[1]中国林业科学研究院森林生态环境与保护研究所,国家林业和草原局森林保护学重点实验室,北京100091;[2]中国热带农业科学院椰子研究所,文昌571339;[3]北京市林业保护站,北京100029;[4]北京农学院植物科学技术学院,北京102206

年份：2021

卷号：57

期号：11

起止页码：49-58

中文期刊名：林业科学

外文期刊名：Scientia Silvae Sinicae

收录：CSTPCD;;EI(收录号：20220311481606);Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金项目(31370644);国家微生物资源平台(NIMR 2017-7)。

语种：中文

中文关键词：嫁接;抗病性;枣疯病;植原体;16S rDNA;himA及其下游假定蛋白

外文关键词：graft;disease resistance;jujube witches’-broom disease;phytoplasma;16S rDNA;himA and its downstream hypothetical protein

分类号：S763.7

摘要：【目的】检测3个枣园不同抗性枣树材料嫁接传毒后携带植原体状况,分析不同植原体株系传播流行、遗传变异和系统发育及其与枣树品种抗性之间的关系,揭示枣疯植原体群体遗传变异特性,为抗病品系合理利用与病害防控提供依据。【方法】4月采集不同抗性枣树材料嫁接到北京玉泉山枣树资源圃、北京昌平王家园和河北唐县军城镇三地枣园的发病砧木上,当年8—10月分别提取接穗和砧木组织样品总DNA,利用植原体16S rDNA、16S-23S间区(SR)、secA、tuf、fusA-tuf间区、himA及其下游假定蛋白基因等片段进行PCR和LAMP检测,并对PCR产物进行测序,分析遗传变异和系统进化。【结果】抗病品系的接穗材料症状严重度大都低于感病砧木;表现无症接穗T30通过直接和巢式PCR与LAMP均未检测到植原体;3个枣园枣树样品中的植原体16S rDNA、SR、tuf和secA基因序列同源性为100%;根据所测38份样品中himA及其下游假定蛋白基因序列在117、154和190 bp位点的碱基变异,军城镇样品序列聚于T-A型,玉泉山样品和河北唐县梁家沟枣疯序列聚于C-A型,而昌平样品和泗洪、泰安、通州枣疯序列聚于C-C型;根据所测29份样品中fusA-tuf基因间区序列在第39 bp位置有碱基A插入或缺失,将玉泉山和军城镇样品归于TJ2型,而国内其他地区各株系和昌平样品为TJ1型;himA及其下游假定蛋白基因序列及fusA-tuf基因间区序列在3个枣园各有优势菌株序列,序列分型对应且互相佐证。【结论】本研究首次测定枣树‘骏优2号’(Z18)优良品系具有高抗病性,并首次检测到3个枣园内枣疯植原体株系的himA及其下游假定蛋白基因与fusA-tuf基因间区存在明显地域差异,且与样品抗病性强弱无关联;初步排除玉泉山枣园植原体来源于江苏泗洪大枣移栽苗传病的假设。
【Objective】This study aimed to detect the jujube witches’-broom(JWB)phytoplasma status of jujubes cultivated in three jujube orchards after inoculating the trees by graft,and to analyze the transmission,epidemic,heritable variation,and phylogeny of different phytoplasma strains;and reveal the relationship between different phytoplasma strains and different resistance jujube cultivars.【Method】In April 2017,different resistant jujube scions were collected and grafted on the phytoplasma-infected rootstocks cultivated in three jujube orchards including Yuquanshan and Changping orchards in Beijing and Juncheng orchard in Tang county,Hebei province.The total DNA of scion and rootstock tissue samples were extracted from August to October of that year,to be used as templates for PCR amplification.Specific PCR products of 16S rDNA,16S-23S intergenic region(SR),secA,tuf,fusA-tuf intergenic region,himA and its downstream putative protein gene were sequenced.Genetic variation and phylogenetic evolution were analyzed with obtained sequence data.【Result】The symptom severity of resistant scions was generally lower than that of susceptible rootstocks.No phytoplasma was detected in all of T30 scions without symptoms of witches’-broom by direct and nested PCR and LAMP.The homology of 16S rDNA,SR,tuf and secA gene sequences of JWB phytoplasma isolates from three orchards was 100%.According to the base variation of himA and its downstream putative protein gene at 117,154 and 190 bp sites in 38 samples,JWB strains from Junchengzhen were clustered in the T-A type,JWB strains from Yuquanshan and Liangjiagou were clustered in the C-A type,while JWB strains from Changping were clustered in the C-C type.According to the insertion or deletion of base A at 39 bp in the sequence of the fusA-tuf intergenic region in 29 samples,JWB strains from Yuquanshan and Juncheng were clustered in the TJ2(A+)type,while others were clustered in the TJ1(A-)type.There were dominant strains in each orchard that cause the sequence types of both regions(himA and its downstream putative protein gene,intergenic region of fusA-tuf genes)were mutually corroborated.【Conclusion】In this study‘Junyou No.2’(Z18)jujube is for the first time identified as the high resistant cultivar to JWB.The variations of different JWB strains from two orchards have no significant correlation with the disease resistance of cultivars,while with the regional difference.The gene sequences of JWB strains from the two regions provide the evidence that JWB strains of Yuquanshan are not originated from Sihong County,Jiangsu Province.The results are helpful to reveal the genetic variation characteristics of JWB phytoplasma population and provide a theory basis for rational utilization of resistant cultivars and JWB disease control.