China has the largest area of kiwifruit production in the world. Pathogens associated with root diseases of kiwi trees have not been investigated extensively. In this research, three Phytophthora species including P. cactorum, P. cinnamomi, and P. lateralis, which are pathogenic to kiwi trees in the main planting areas of China, were studied. The population densities of these species in 128 soil samples from 32 kiwi orchards in 2017 and 2018 were measured using multiplex real-time quantitative PCR, based on the ras-related protein gene Ypt1. P. cactorum was the most widely distributed of the three species in orchards of the Zhouzhi and Meixian Prefectures. We also used redundancy analysis (RDA) to examine soil factors in the kiwi orchards, to understand their effects on the population densities of the Phytophthora species. The RDA analysis indicated that soil temperature and pH were significantly correlated with the abundance of P. cactorum and P. cinnamomi. Besides, two loop-mediated isothermal amplification (LAMP) detection systems for P. cactorum were developed based on the tigA gene. The color-change detection system was proved to be accurate, sensitive, and faster than quantitative PCR. The results of this study, along with the LAMP detection systems, will be of great use in the control of Phytophthora diseases for the production of kiwifruits in China.
Aconitum carmichaelii Debeaux is a traditional Chinese medicinal herb that has been utilized for approximately two thousand years. However, as cultivation has increased, there have been more reports of A. carmichaelii infections caused by four major pathogenic fungal species, Fusarium oxysporum, F. solani, Mucor circinelloides, and Sclerotium rolfsii, resulting in increased disease incidences and limited production and quality. To detect these infections, we developed a LAMP-based toolbox in this study. The cytochrome c oxidase subunit 1 (cox1) gene, translation elongation factor-1α (EF-1α), internal transcribed spacer (ITS) regions of rDNA, and alcohol dehydrogenase 1 (ADH1) gene, respectively, were used to design species-specific LAMP primer sets for F. oxysporum, F. solani, S. rolfsii, and M. circinelloides. The results showed that the LAMP-based toolbox was effective at detecting pathogens in soil and plant materials. We also used this toolbox to investigate pathogen infection in the main planting regions of A. carmichaelii. Before harvesting, F. oxysporum, M. circinelloides, and S. rolfsii, were commonly found in the planting fields and in infected A. carmichaelii plants. Therefore, the toolbox we developed will be useful for tracking these infections, as well as for disease control in A. carmichaelii.
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