Establishment and application of a rapid high-throughput detection method for Huanglongbing.
10.19540/j.cnki.cjcmm.20250122.101
- Author:
Qin YUAN
1
;
Zhi-Peng LI
1
;
Tie-Lin WANG
2
;
Ting DONG
1
;
Yu-Wen YANG
1
;
Wei GUAN
1
;
Ting-Chang ZHAO
1
Author Information
1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences Beijing 100193, China.
2. State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
- Publication Type:English Abstract
- Keywords:
Citri Reticulatae Pericarpium;
Huanglongbing;
TaqMan qPCR detection;
alkali lysis method
- MeSH:
Citrus/microbiology*;
Plant Diseases/microbiology*;
Rhizobiaceae/physiology*;
High-Throughput Screening Assays/methods*;
Liberibacter/physiology*
- From:
China Journal of Chinese Materia Medica
2025;50(7):1735-1740
- CountryChina
- Language:Chinese
-
Abstract:
The dried mature peel of Citrus reticulata, a plant in the Rutaceae family and its cultivated varieties, is a commonly used Chinese medicinal material known as Chenpi(Citri Reticulatae Pericarpium). It is rich in nutritional components and medicinal value, with pharmacological effects including relieving cough and eliminating phlegm, strengthening the spleen and drying dampness, protecting the liver and benefiting the stomach, tonifying Qi, and calming the mind. Huanglongbing(HLB), also known as Citrus Huanglongbing, is a destructive disease in citrus production that seriously threatens the development of the citrus industry. HLB causes symptoms such as the inability of Rutaceae plants to produce mature fruit, gradual weakening of the tree, and eventual death, posing a significant threat to the yield and quality of Chenpi. Due to the uneven distribution of the HLB pathogen in infected plants, accurate detection of the pathogen requires the collection of a large number of plant samples. Current sample pretreatment methods, such as traditional extraction methods and commercial extraction kits, are time-consuming and involve multiple steps, which significantly increase the difficulty and workload of HLB diagnosis and have become a bottleneck in HLB detection. In this study, a rapid high-throughput detection method combining alkali lysis and TaqMan qPCR was developed. This method allows the pretreatment of multiple samples within 5 min, and the entire detection process can be completed within 45 min, with a detection limit of 6.67 fg·μL~(-1). The alkali lysis method and commercial kits were used for parallel detection of field-collected citrus samples, and the results showed no significant difference. The sample pretreatment method established in this study is characterized by low cost, simplicity, and high efficiency. Combined with TaqMan qPCR, it can provide technical support for early and on-site diagnosis of HLB. This method is of great significance for disease prevention and control in the citrus industry and is expected to help improve the yield and quality of citrus medicinal materials.
