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• W2355216919 abstract "Cotton seeds collected from plants infected by Verticillium ablo-atrum were used for the purpose of studying seed transmission of the pathogen. Although verticilliate conidiophores had been observed occasionally under low-power microscope on the surface of the seeds plated on PDA in 1955, yet isolations of the organism failed due to the interference of several contaminants. Seeds were delinted in order to diminish the possibility of interference from lint-born contaminants. The materials were plated on different culture media such as PDA, acidified PDA, cotton meal agar and rice agar. All these plates failed to show verticillium colonies. Among the fungi obtained in these plates, Fusarium, Alternaria and Colletotrichum were the most frequent. Since both Verticillium albo-atrum and other fungi were harbored within the same seed, the difference of growth rates might be considered as an important factor involved in the difficulty of the isolations: Cultures of V. albo-atrum and Fusarium sp. were transferred simutaneously to PDA plates to test their growth rates. The diameter of their colonies were measured after incubation for 12 days under 22±1℃. Those of V. albo-atrum measured 2.8 cm., while the average of the latter was 9,3 cm. Interference of the fast-growers might render a slow-grower such as V. albo-atrum to be masked in the observations and to be depressed in the isolations. A method was developed to avoid the interference. The fundamental procedures of the method are outlined as follows. 1. Plating and observation. (1) Cotton seeds being delinted with commercial surphuric acid in order to avoid the interference from lint-born contaminants. (2) Delinted seeds being washed in a flask covered with wire gauze for 24 hours under running tap water in order to get rid of bacteria. Absorption of sufficient water during washing activates the hibernating mycelium. (3) Seeds being plated on 1.7% water agar instead of nutrient agar in order to diminish the growth of the fast-growers such as Fusaria and Alternaria. The plates being incubated under 22±1℃. for 15 days. (4) The plates being uncovered and observed directly under low-power microscope in order to observe the fungi undisturbed. 2. Isolation and preservation of the Verticillium cultures. (1) Verticilliate conidiophores being located under low-power microscope. (2) The tip of a flamed needle moistened with sterile agar being introduced within the microscopic field in order to fish the spores under focus. (3) Spore suspension being made by washing the needle in a 5 cc sterile water blank. (4) Dilution plates being made by placing 1 cc of the spore suspension in a petri dish and diluting with 10 cc of cotton stem agar. (5) The plates being observed under low-power microscope after incubation for 5—7 days at 22±1℃. (6) Verticillium colonies being locatad and a bit of the margin of a colony being transferred to PDA slant. (7) The slant cultures being flooded with sterile liquid paraffin after incubation for 7—15 days at 22±1℃. The cultures preserved with liquid paraffin were able to keep alived for 2 years. Seeds collected from the infected plants in 1955, 1956 and 1957 were studied. The percentages of seeds carrying Verticillium observed in these years were 39.8, 5.92 and 9.5% respectively. One of the representative single spore colonies obtained was identified as V. albo—atrum. Inoculation and reisolation tests showed positive results. Inspection of cotton seed samples collected from eight co-op farms in Shensi Province in 1958 showing that the percentages of seeds infected by verticillium ranged 3 to 23% with an average of 12.3%." @default.
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• W2355216919 date "1958-01-01" @default.
• W2355216919 modified "2023-09-23" @default.
• W2355216919 title "A METHOD FOR THE INSPECTION AND ISOLATION OF SEED-BORN VERTICILLIUM FROM COTTON SEEDS" @default.
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