Ustilaginoidea virens spore real-time quantification loop-mediated isothermal amplication detection method and kit

Abstract

The invention discloses a LAMP-specific primer composition used in a q-LAMP rapid detection method for rice smut spores. The LAMP-specific primer composition is composed of four primers: upstream and downstream outer primers and upstream and downstream inner primers. The present invention also discloses a loop-mediated isothermal amplification kit for the quantitative detection of rice smut spores, which comprises the above-mentioned LAMP-specific primer composition, and also contains betaine, 10×SYBR Green I, 8U / μL Bst DNA polymerase, etc. The invention also correspondingly discloses a loop-mediated isothermal amplification method for quantitatively detecting the spores of the rice false smut fungus.

Description

technical field [0001] The invention relates to a real-time quantitative loop-mediated isothermal amplification (q-LAMP) detection method and kit for rice false smut, belonging to the technical field of early warning of plant disease detection, identification and prevention. Background technique [0002] Rice false smut is a worldwide rice disease caused by Ustilaginoidea virens (Cooke) Tak., which is seriously harmful in China, Japan, India and other countries. The disease is an ear disease, and white rice balls are formed on the ears at the beginning, and then quickly turn into yellow or dark green rice balls. The disease significantly affects rice yield and rice quality, and the annual yield loss rate is 30-50% in severe cases, and the processed rice is mixed with a large number of chlamydospores. Chlamydospores of rice koji fungus contain the mycotoxin Ustilaxin. The results of animal toxicity experiments by Japanese researchers showed that chlamydospore water extract ...

AI Technical Summary

Problems solved by technology

[0003] Rice false smut is one of the most difficult pests of rice to control at present. The main reason is that it is difficult to grasp its infection dynamics in a timely and rapid manner with existing technologies.

This leads to many agents having high inhibitory activity against rice smut, but the effect is not good in actual application

The main reason is that due to the lack of sensitive and fast bacteria monitoring technology, it is impossible to achieve scientific drug use and scientific prevention

At present, although the number of overwintering chlamydospores in the soil can be checked by artificial isolation culture and conventional microscope observation; however, there are many kinds of microorganisms in the paddy field, and the growth rate of rice mistletoe is slow, and the hyphae and conidia produced by the germination of chlamydospores Without specific morphological characteristics, it is difficult to identify by pathological means such as microscope observation and isolation culture, and it is impossible to understand the changes in the number of rice false smut bacteria in the field during the rice growth season, the infection of plants and their proliferation in plants. These monitoring techniques It takes more than a week

[0004] With the continuous development of nucleic acid-related identification methods, the method based on common PCR has been successfully used for the detection of pathogenic bacteria, but the specificity of the PCR method takes a long time, requiring 6-8 hours, and the reaction is easily affected by PCR inhibitors. The pre-processing process is complicated

At the same time, the PCR method requires an expensive instrument PCR machine, and the detection sensitivity is low, and the detection process is complicated

Images