Abstract
Simple SummaryIn this study, we investigated the soft rot disease of Dendrobium officinale in Guizhou Province, and designed a rapid method to detect the pathogen is Fusarium oxysporum in Dendrobium officinale by using specific gene sequences and loop-mediated isothermal amplification technology. This method can accurately detect Fusarium oxysporum on soft rot plants of Dendrobium officinale. Therefore, the results of this study are crucial for the early diagnosis of soft rot on Dendrobium officinale.Soft rot causing Fusarium oxysporum is one of the most destructive diseases of Dendrobium officinale Kimura et Migo in China that reduces D. officinale yield and quality. A key challenge for an integrated management strategy for this disease is the rapid and accurate detection of F. oxysporum on D. officinale. Therefore, a new loop-mediated isothermal amplification (LAMP) assay was developed for this purpose. In this study, the primers were selected and designed using the translation elongation factor-1α (TEF-1α) gene region as the target DNA sequence in order to screen the best system of reaction of LAMP to detect F. oxysporum through optimizing different conditions of the LAMP reaction, including time, temperature, concentrations of MgSO4, and concentrations of inner and outer primers. The optimized system was able to efficiently amplify the target gene at 62 °C for 60 min with 1.2 μM internal primers, 0.4 μM external primers, 7 mM Mg2+, and 5 fg/µL minimum detection concentration of DNA for F. oxysporum. The amplified products could be detected with the naked eye after completion of the reaction with SYBR green I. We were better able to control the effect of soft rot in D. officinale using fungicides following a positive test result. Additionally, the control effect of synergism combinations against soft rot was higher than 75%. Thus, LAMP assays could detect F. oxysporum in infected tissues of D. officinale and soils in field, allowing for early diagnosis of the disease.
Highlights
Dendrobium officinale Kimura et Migo is a perennial herb belonging to the Orchidaceae Dendrobium Sw. that is commonly used in traditional Chinese medicine [1,2]
The colonies of strain XY1E208 were circular on potato dextrose agar (PDA) medium with hyphae stripes
Our study showed that loop-mediated isothermal amplification (LAMP) assays could correctly detect F. oxysporum
Summary
Dendrobium officinale Kimura et Migo is a perennial herb belonging to the Orchidaceae Dendrobium Sw. that is commonly used in traditional Chinese medicine [1,2]. Most studies have shown that D. officinale contains abundant mineral elements, amino acids, and fatty acids, which explains its high medicinal value [3,4]. D. officinale is distributed in east Asia, southeast Asia, and Australia, along with a few other countries or regions. It is mainly distributed in the southern subtropical areas of China [7,8]. D. officinale requires a very specific growth environment and particular climactic conditions. It grows slowly so the wild resources are very scarce [9,10]. With increasing market demand for D. officinale, people often choose to imitate the wild cultivation of D. officinale [11,12]
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