Real-time PCR for detection and quantification of C. gloeosporioides s.l. growth in Stylosanthes and Arabidopsis
Real-time PCR for detection and quantification of C. gloeosporioides s.l. growth in Stylosanthes and Arabidopsis
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用于检测和定量 C. gloeosporioides s.l. 的实时 PCR
DOI：
10.1016/j.cropro.2022.106021
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发表时间：
2022-05
期刊：
影响因子：
2.8
通讯作者：
Lijuan Luo
中科院分区：
文献类型：
--
作者：
Miting Wan;Liyun Yang;Shizi Zhang;Jing Gao;Lingyan Jiang;Lijuan Luo
Colletotrichum gloeosporioides sensu lato (s.l.) 是经济和科学上最重要的真菌病原体之一，在热带和亚热带地区引起严重疾病。 C. gloeosporioides s.l. 的检测对疾病控制具有重要意义。在本研究中，开发了一种基于 ACT 的实时 PCR 测定法，用于对 C. gloeosporioides s.l. 进行定量，该测定法可靠地检测到少至 100 fg 基因组 DNA、100 个目标 DNA 拷贝和 20 个分生孢子。该方法可以识别所有四种测试的 C. gloeosporioides s.l。分离株，而在其他炭疽菌属物种和灰葡萄孢中没有观察到扩增，表明该测定的特异性。 C. gloeosporioides s.l. 的检测和定量在人工和自然感染的宿主叶子中得到了证明。首先，使用接种后不同时间点收集的叶子样品进行实时 PCR 分析，以监测 C. gloeosporioides s.l 的生长。随着时间的推移。其次，利用该方法比较了两个柱花品种和两个拟南芥品种的抗性。最后采用实时荧光定量PCR方法对田间自然感染和无症状的柱花草叶片进行检测。总体而言，实时 PCR 检测可以在感染的早期无症状阶段进行检测；并且结果与显微镜观察和后期疾病症状有很好的相关性。因此，我们的研究为研究植物-炭疽菌相互作用以及疾病预测和控制提供了一种快速有效的检测方法。 • 建立了炭疽杆菌的检测方法。 • 方法的特异性和敏感性。 • 适用于柱花属和拟南芥属植物的检测。 • 适用于自然感染宿主样本。
Colletotrichum gloeosporioides sensu lato (s.l.) is one of the most economically and scientifically important fungal pathogens, causing serious diseases in tropical and subtropical region. The detection of C. gloeosporioides s.l. is of importance for disease control. In this study, an ACT-based real-time PCR assay was developed for quantification of C. gloeosporioides s.l., which reliably detected as little as 100 fg genomic DNA , 100 copies of target DNA and 20 conidia . This method could recognize all four tested C. gloeosporioides s.l. isolates, while no amplification was observed in other Colletotrichum species and Botrytis cinerea , indicating the specificity of this assay. Detection and quantification of C. gloeosporioides s.l. was demonstrated in artificially and naturally infected host leaves. First, the real-time PCR analysis was performed using leaf samples collected at different time points post inoculation to monitor the growth of C. gloeosporioides s.l. over time. Secondly, the method was used to compare the resistance in two Stylosanthes cultivars and two Arabidopsis cultivars. Finally, naturally infected and symptomless leaves of Stylosanthes in the fields were tested by the real-time PCR method. Overall, the real-time PCR assay could allow the detection at early symptomless phase of infection; and the results was well correlated with microscopic observation and later disease symptoms. Therefore, our studies have provided a rapid and effective detection method for studying the plant- Colletotrichum interaction as well as disease prediction and control. • Established a detection method for C. gloeosporioides. • The specificity and sensitivity of the method. • Suitable for the detection in Stylosanthes and Arabidopsis. • Suitable for the natural infected host samples.
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影响因子：
5.1
作者：
Ross A;Somssich IE
通讯作者：
Somssich IE
影响因子：
5.1
作者：
Weßling R;Panstruga R
通讯作者：
Panstruga R
影响因子：
--
作者：
Liu Guodao;C. Phaikaew;W. Stür
通讯作者：
Liu Guodao;C. Phaikaew;W. Stür
DOI：
10.1016/j.plaphy.2017.06.024
发表时间：
2017-09
期刊：
Plant physiology and biochemistry : PPB
影响因子：
--
作者：
Hui Wang;Zhijian Chen;Guodao Liu;C. Bai;H. Qiu;Yan-An Jia;Lijuan Luo
通讯作者：
Hui Wang;Zhijian Chen;Guodao Liu;C. Bai;H. Qiu;Yan-An Jia;Lijuan Luo
影响因子：
0.7
作者：
A. Chandra
通讯作者：
A. Chandra