1997
DOI: 10.1111/j.1439-0434.1997.tb00410.x
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Preparation and Regeneration of Mycelial Protoplasts of Alternaria eichhomiae

Abstract: Preparation and regeneration of mycelial protoplasts from Aliernaria eichhorniae were examined. A commercially available muralytic enzyme, Novozym 234, was used for isolation of protoplasts. The mycelial age and the pH of the stabilized buffer affected the formation of protoplasts. The maximum production of protoplasts (3,9 X 10*/gfresh weight mycelia) was obtained from 24-h-old mycelia digested with Novozym 234 (20 mg/m!) in a stabilized buffer of pH 6.4 and incubated in the dark at 30"C on a rotary shaker (9… Show more

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Cited by 6 publications
(5 citation statements)
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“…For V. dahliae , previous studies have used 24-h-old mycelia for protoplast isolation [ 18 , 19 ], but here we obtained more protoplasts from the 20-h culture than from the 24-h culture. The probable reason for this can be the sensitivity of the younger mycelia to the digesting enzymes, with the increase in growth the cell wall becomes thicker and the mycelia would be digested more difficultly leading to decreased protoplast yield [ 36 , 38 , 39 ]. In addition to the age of mycelia, the protoplast yield also depends on the duration of the enzyme digestion, which has ranged from 3 h to 16 h for maximum protoplast release in other fungal species [ 37 , 40 , 42 ].…”
Section: Discussionmentioning
confidence: 99%
“…For V. dahliae , previous studies have used 24-h-old mycelia for protoplast isolation [ 18 , 19 ], but here we obtained more protoplasts from the 20-h culture than from the 24-h culture. The probable reason for this can be the sensitivity of the younger mycelia to the digesting enzymes, with the increase in growth the cell wall becomes thicker and the mycelia would be digested more difficultly leading to decreased protoplast yield [ 36 , 38 , 39 ]. In addition to the age of mycelia, the protoplast yield also depends on the duration of the enzyme digestion, which has ranged from 3 h to 16 h for maximum protoplast release in other fungal species [ 37 , 40 , 42 ].…”
Section: Discussionmentioning
confidence: 99%
“…TES and TES1 null mutants were generated by disrupting the targeted gene using homologous recombination, respectively ( Figure 3 A, C). Protoplasts were prepared from A. alternata ZJ33 as described previously with modifications [ 14 , 34 ]. A. alternata ZJ33 was cultured in YPG liquid medium at 28 °C for 48 h in a rotary shaker (180 rpm), and mycelia were harvested.…”
Section: Methodsmentioning
confidence: 99%
“…The physiological stage of the mycelium plays a significant role in the release of protoplasts [ 62 , 63 ]. Here, the culture medium used had a great influence on the growth of A. niger .…”
Section: Discussionmentioning
confidence: 99%
“…For instance, A. niger conidia were cultured in 100 mL complete medium (CM) for 12 h (at 100 rpm) to generate protoplasts [ 14 ]. It seems that young mycelia are more susceptible to enzymatic digestion than old mycelia and that the cell wall of old mycelia becomes thicker [ 22 , 63 , 64 , 65 ].…”
Section: Discussionmentioning
confidence: 99%