A Simple and Rapid Method for Isolating Monospores of Cercospora

Abstract

The invention relates to a method for simply and quickly separating cercospora monospores. A specially-made spore taking device is adopted, one end of the spore taking device is made into a flat round spore taking ball, and the other end of the spore taking device is made into a spore taking shovel which is slightly sharp at the front end and is in a flat shovel shape. The spore taking device can glue pathogen spores of a small amount, a culture medium is evenly coated with the pathogen spores, it can be guaranteed that one spore arises in a view of 10*10 times, the spore density is low, a target spore is easy to find, blade residual bodies do not adhere easily, the pathogen spores can be separated easily, and the separation rate and separation speed are improved effectively. The chosen spore is a spore which is cultured for 24 h and is germinated, the result that germination cannot occur after single spores are chosen in other methods is effectively avoided, and the success rate is high. The spore is chosen in a mode of spore taking through adhesion, the operation is quick, the success rate is high, the picking efficiency is improved, the operation technique difficulty of the method is not high, the method can be mastered skillfully after several trainings, and the method is easy to handle, simple and easy to learn.

Description

technical field [0001] The invention relates to a simple and rapid method for isolating single spores of the genus Cercospora, which belongs to the technical field of microorganism culture. Background technique [0002] Cercospora fungi are plant pathogenic bacteria that can parasitize on the stems, leaves, flowers or fruits of plants and cause diseases. Cercospora zeina (Meisel B, Korsman J, Kloppers F J, et al. Cercospora zeina is the causal agent of gray leaf spot disease of maize in southern Africa[J]. European journal of plant pathology, 2009, 124(4 ):577-583.), sugar beet brown spot (Cercospora beticola) (Holtschulte B, Asher M J C, Molard M R, et al. Cercospora beticola-worldwide distribution and incidence[J]. sugar beet.,2000:5-16.), pepper brown spot (Cercospora capsici) (Kawagoe H.Influence of temperature and relative humidity on disease occurrence of sweet pepper caused by Cercospora capsici in a plastic house[J].Annals of the Phytopathological Society ofJapan, ...

AI Technical Summary

Problems solved by technology

[0004] Wherein, "single-spore separator" is a single-spore isolation device (Constantinescu O.An instrument and procedure for single-spore isolation[J].Transactions of the British Mycological Society, 1988,91(4):700-702.), However, the installation of this device is relatively complicated, and the microscope equipment needs to be modified, and this device has not been widely used in China.

In contrast, manual separation is more common, but different methods also have some shortcomings, for example, the cutter separation method (Ho W C, Ko W H.A simple method for obtaining single-spore isolates of fungi[J].Botanical Bulletin of Academia Sinica , 1997, 38.), it is necessary to mark the single spores under the microscope, and then cut the single spore agar block with a cutter, often the range of cutting is not accurate, and the situation of single spores cannot be cut; directly pick the spores under the microscope There are agar plates with spores, but the petri dish can not be fixed, nor can it be moved arbitrarily, causing the separation rate to decrease (Fang Zhongda. Plant Disease Research Methods [M]. Beijing: Agricultural Press, 1998.); Glass needle method (Choi Y W , Hyde K D, Ho W H.Single spore isolation offungi.Fungal Diversity,1999,3:29-38.), eyebrow acupuncture method (Liu Changhua, Wang Zhenhua. Study on the relationship between corn head smut inoculation concentration and incidence in the field[J ]. Corn Science, 2008,16 (1): 119-121.), these methods basically all are to pick spores by picking needles, and the tools for making picking needles are mostly insect needles or glass picking needles, and insect needles are thinner. It is easy to determine the location of the spores under a microscope, but the insect needle is soft, and it is difficult to pick out the agar block with spores stuck, while the glass pick needle has moderate hardness, but its diameter is relatively thick, and the observation under the microscope often occupies the entire field of view, and the spores cannot be identified At the same time, the tip of the glass pick is fragile, so be careful when handling it

These factors reduce the success rate and operational efficiency of single spore isolation, and limit the application of single spore isolation method to a certain extent.

Images