Periodic selection in longterm continuous-flow cultures of the filamentous fungus Fusarium graminearum.

M G Wiebe, G D Robson, B Cunliffe, S G Oliver, A P Trinci
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引用次数: 20

Abstract

By monitoring increases and decreases in the proportion of cycloheximide-resistant macroconidia, periodic selection was observed in populations of the filamentous fungus Fusarium graminearum, grown in glucose-limited chemostat cultures. The results indicated that periodic selection of advantageous mutants of F. graminearum occurred at intervals of about 124 h at both high (D = 0.19 h-1, approximately 34 generations) and low (D = 0.06 h-1, approximately 11 generations) dilution rates. Several 'adaptive' peaks (each indicating the appearance of an advantageous mutation) were observed before morphological (highly branched) mutants appeared in the populations; these mutants have previously been observed to have a selective advantage over the parental strain. At intervals, macroconidia harvested from the chemostat were used to inoculate plates of non-antibiotic-containing agar medium, and it was possible to monitor periodic selection in the original chemostat culture using second generation macroconidia harvested from these cultures. The proportion of cycloheximide-, potassium chlorate-, and p-fluoro-DL-phenylalanine-resistant macroconidia in these second generation macroconidia changed in a pattern similar to that observed when monitoring the proportion of cycloheximide-resistant macroconidia in the first generation population harvested directly from the chemostat. The experiments demonstrated that populations of filamentous fungi are heterogeneous and that much of this heterogeneity may already be present at the end of batch growth, i.e., before the onset of continuous cultivation.

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丝状真菌谷草镰刀菌长期连续培养中的周期性选择。
通过监测抗环己亚胺大分生孢子比例的增加和减少,观察到在葡萄糖限制化培养物中生长的丝状真菌镰刀菌(Fusarium graminearum)群体的周期性选择。结果表明,在高稀释率(D = 0.19 h-1,约34代)和低稀释率(D = 0.06 h-1,约11代)下,有利突变体的周期性选择间隔约124 h。在种群中出现形态(高度分支)突变之前,观察到几个“适应性”峰值(每个峰值表明有利突变的出现);这些突变体先前已被观察到比亲本菌株具有选择优势。每隔一段时间,从恒化菌中收获的大分生孢子被用于接种不含抗生素的琼脂培养基,并且可以使用从这些培养基中收获的第二代大分生孢子来监测原始恒化菌培养中的周期性选择。第二代大分生孢子中耐环己亚胺、氯酸钾和对氟dl -苯丙氨酸的大分生孢子比例的变化模式与监测直接从趋化器中收获的第一代大分生孢子中耐环己亚胺的大分生孢子比例的变化模式相似。实验表明,丝状真菌的种群是异质的,而且这种异质的大部分可能在批量生长结束时就已经存在,即在连续培养开始之前。
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