Method for detecting fungi carried by rice seeds using improved Bengal red medium
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A technology of Bengal red and rice seeds, which is applied in microorganism-based methods, biochemical equipment and methods, and determination/inspection of microorganisms, etc. Effects of bacterial contamination
Active Publication Date: 2019-12-06
YUNNAN LINPENG AGRI SCI & TECH CO LTD
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[0010] The technical problem to be solved by the present invention is to overcome the existence of isolated fungi in the prior art of seed health detection, which are susceptible to bacterial interference, so that there are few types of isolated fungi, which cannot fully reflect the fungi carried by the seeds, and when separated, some fungi grow faster. Fast, long colony diameter, interference to the isolation of other fungi, and the defect that it is difficult to isolate fungi alone
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Embodiment 1
[0027] The inventive method, the steps are as follows:
[0028] (1) Improve the preparation of Bengal red culture medium, weigh the ingredients according to the improved Bengal red culture medium formula, put them in a conical flask, add 1L of water, and sterilize at 121°C, 0.15MPa, moist heat for 20min according to the moist heat sterilization method , sub-packed into 9cm diameter petri dish for subsequent use (can stay in the triangular flask as not being used for the time being, sub-package after next time sterilization); The formula of described improved Bengal red culture medium is: add peptone ( Peptone) 5g, glucose (Glucose) 10g, potassium dihydrogen phosphate (KH 2 PO 4 )1g, MgSO 4 ·7H 2 O 0.25g, Agar powder 15g, Bengal red (Tetrachlorofluorescein) 0.033g, chloramphenicol (Chloramphenicol) 0.1g, copper sulfate (CuSO 4 ) 0.05g, zinc nitrate (Zn(NO 3 ) 2 ) 0.029g, cadaverine (1,5-Pentanediamine) 0.0001g;
[0029] (2) After the improved Bengal red culture medium in...
Embodiment 2
[0036] Embodiment 2 except being the Yuanyang red rice (Moon Valley) rice seed that gets in addition, all the other experimental methods are identical with embodiment 1, and experimental effect sees for details figure 2 and Figure 4 .
[0037] figure 2 Among them, 2 kinds of fungi were detected in the culture medium of the present invention, ( figure 2 As shown in A, figure 2 In A, 1 is a fungus of the genus Penicillium (Penicillium), and 2 is a fungus of the genus Pythium. See the picture of the spores of the isolated fungi Figure 4 ), the diameter of the fungal colony is 2-3cm, which is convenient for picking and observing. Two kinds of fungi were detected in PDA medium ( figure 2 As shown in B, figure 2 In B, 3 is a fungus of the genus Pythium, and 4 is a fungus of the genus Penicillium. See the picture of the spores of the isolated fungi Figure 4 ), the fungal colonies almost covered the petri dish, the growth of the two fungi overlapped, and it was difficu...
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Abstract
The invention discloses a method for detecting fungi carried by rice seeds by using an improved rose Bengal culture medium. The method comprises the steps of culturing and detecting the rice seeds by using the improved rose Bengal culture medium, wherein the culturing comprises the steps of putting culture dishes in an incubator, carrying out culturing for 1 day under the conditions of 25 DEG C, 9,000Lx illumination intensity, 12h illumination and 12h darkness, putting the culture dishes in a refrigerator with the temperature of -20 DEG C after the seeds are imbibed, keeping the temperature for 12 hours, taking out the culture dishes, carrying out culturing for 6 days under the conditions that the constant temperature is 28 DEG C, the illumination intensity is 9,000Lx and the light cycle is 12h light period and 12h dark period, picking the fungi, and microscopically observing and checking species of the fungi carried by the seeds. According to the improved rose Bengal culture medium, copper sulfate, zinc nitrate and cadaverine are added, and the consumption of magnesium sulfate is lowered. The method disclosed by the invention is beneficial to the growth of heterogeneous fungi, the species of the fungi can be detected and separated, the operation is not prone to bacterial contamination, the diameter of fungal colonies is relatively small, and thus, the separation and counting of the fungi are facilitated.
Description
technical field [0001] The invention belongs to the technical field of seed quality detection, and in particular relates to a method for detecting fungi carried by seeds. Background technique [0002] At present, seed health testing has become a routine measure in the process of seed "quality assessment" and "phytosanitary inspection" in many countries, and has become an indispensable part of the seed quality control system. Some countries have established special seed health inspections. Assay laboratory. Seed health measurement is to determine whether the seeds carry pathogenic bacteria (such as fungi, bacteria and viruses), harmful animals (such as nematodes and pests) and other health conditions, that is, to detect the types and quantities of diseases and insect pests carried by seeds, and the detection of fungi carried by seeds It is an important content in the detection of seed health. For example, since the establishment of the Seed Health Unit (SEED HEALTH UNIT) in...
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