核桃细菌性黑斑病菌LUX发光标记菌株的构建及其在核桃抗性评价中的应用 Click here to download the full text Citation：徐海娇,崔建潮,赵文诗,于秋香,许长新,王新茹,李立涛,焦蕊,贺丽敏.核桃细菌性黑斑病菌LUX发光标记菌株的构建及其在核桃抗性评价中的应用.Journal of Plant Protection,2025,52(2):316-326 Hits: Download times: Author Name Affiliation E-mail Xu Haijiao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Cui Jianchao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Zhao Wenshi  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Yu Qiuxiang  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Xu Changxin  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Wang Xinru  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Li Litao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Jiao Rui  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    He Limin  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China  helimin122@163.com  中文摘要:为便于核桃细菌性黑斑病菌黄单胞杆菌Xanthomonas arboricola pv.juglandis的早期定量检测、侵染过程观察以及核桃品种的抗性评价，将发光杆菌Photorhabdus luminescens的luxCDABE操纵子转入到黄单胞杆菌野生型菌株Xaj细胞中，构建了组成性表达lux基因的黄单胞杆菌发光标记菌株Xaj-LUX，测定该菌株中LUX光信号的稳定性及其生长曲线和致病性，并对核桃叶片中菌株Xaj-LUX发光值与黄单胞杆菌浓度进行相关分析，基于此对71份核桃种质资源进行抗性评价。结果显示： luxCDABE的转入不影响黄单胞杆菌野生型菌株Xaj的菌落形态、生长及致病力；在侵染核桃叶片过程中，菌株Xaj-LUX发光值与黄单胞杆菌浓度呈显著线性相关，表明发光值能准确反映叶片中黄单胞杆菌浓度；在71份核桃种质资源中，高抗种质资源17份，中抗种质资源23份，低抗种质资源18份，分别占供试总数的23.9%、32.4%和25.4%，而中感和高感种质资源所占比例分别为11.3%和7.0%。表明菌株Xaj-LUX可用于核桃细菌性黑斑病菌的快速定量检测及其在植物体内的高通量检测。 中文关键词:核桃  细菌性黑斑病菌  发光标记菌株  生物发光  抗性评价   Construction of a LUX-tagged strain of Xanthomonas arboricola pv. juglandis and its application in the evaluation of walnut disease resistance Author Name Affiliation E-mail Xu Haijiao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Cui Jianchao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Zhao Wenshi  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Yu Qiuxiang  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Xu Changxin  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Wang Xinru  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Li Litao  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    Jiao Rui  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China    He Limin  Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry Sciences, Changli 066600, Hebei Province, China  helimin122@163.com  Abstract:To facilitate in vivo quantitative detection of Xanthomonas arboricola pv. juglandis and observation of the pathogen infection process at the early stage, and the evaluation of disease resistance in walnut varieties, the luxCDABE operon from Photorhabdus luminescens was transformed into the wild type strain Xaj, constructing a constitutively luminescent strain named Xaj-LUX. The stability of luminescence, bacteria growth curve and pathogenicity of the Xaj-LUX strain were measured, and the correlation analysis between the luminescence value and the concentration of strain Xaj-LUX in walnut leaves was performed. Subsequently, evaluation on the resistance of 71 walnut germplasm resources was conducted. The results showed that bacterial colony morphology, growth, or pathogenicity of the wild type strain Xaj were not affected by the integration of luxCDABE. During the infection process of walnut leaves, the luminescence intensity of the strain Xaj-LUX showed a significantly linear correlation with the concentration of X. arboricola pv. juglandis, accurately and exhibited the growth of X. arboricola pv. juglandis in leaves. Among the 71 walnut germplasm resources, 17 showed high resistance, 23 middle resistance and 18 low resistance, accounting for 23.9%, 32.4% and 25.4% of the total numbers of the germplasms tested, respectively; while middle susceptible and high susceptible germplasms accounted for 11.3% and 7.0%, respectively. These findings demonstrate that the Xaj-LUX strain can be used for rapid quantitative detection of the walnut bacterial blight pathogen and highthroughput monitoring within plant tissues. keywords:walnut  Xanthomonas arboricola pv. juglandis  luminescent marker strain  bioluminescence  resistance evaluation View Full Text  View/Add Comment  Download reader

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