Department of Plant Pathology, Departments of Forestry and Statistics, Department of Botany, and Department of Soil Science, University of Wisconsin-Madison, Madison, Wisconsin 53706.
Appl Environ Microbiol. 1988 Jun;54(6):1430-5. doi: 10.1128/aem.54.6.1430-1435.1988.
Colletotrichum gloeosporioides, Colletotrichum coccodes, Colletotrichum phomoides, and Acremonium strictum. When the fungi were grown on solid medium in microplates and spore yields were measured by counting the subsamples with a hemacytometer, the variability among hemacytometer squares was always the largest source of variation, accounting for 51 to 91% of the total variation. Variability among replicate cultures and results of repeat experiments were generally also significant. The effect of square-to-square variability on the precision of spore yield measurement was minimized by counting a moderate number (ca. 30) of squares per culture. Culture-to-culture variability limited the practical precision of spore production measurements to a 95% confidence interval of approximately the mean +/- 25%. We provide guidelines for determining the number of replicate cultures required to attain this or other degrees of precision. Particle counter-derived spore counts and counts based on spore weights were much less variable than were hemacytometer counts, but they did not improve spore production estimates very much because of culture-to-culture variability. Results obtained by both of these methods differed from those obtained with a hemacytometer; particle counter measurements required a correction for spore pairs, while the relationship between spore weights and spore counts changed as the cultures aged.
炭疽菌、胶孢炭疽菌、番茄炭疽菌和严格青霉中,研究了真菌孢子产生的可变性和孢子产量的测量。当真菌在微孔板上的固体培养基中生长,通过血细胞计数器对亚样本进行计数来测量孢子产量时,血细胞计数器方格之间的变异性始终是最大的变异来源,占总变异的 51%至 91%。重复培养之间的变异性和重复实验的结果通常也很显著。通过对每个培养物计数约 30 个方格,可以最大限度地减少方格间变异性对孢子产量测量精度的影响。由于培养物间的变异性,孢子产生测量的实际精度限制在 95%置信区间的平均值正负 25%左右。我们提供了确定达到这种或其他精度所需的重复培养物数量的指南。粒子计数器衍生的孢子计数和基于孢子重量的计数比血细胞计数器计数的变异性小得多,但由于培养物间的变异性,它们并没有很大程度地改善孢子产生的估计。这两种方法的结果与血细胞计数器的结果不同;粒子计数器测量需要对孢子对进行校正,而随着培养物的老化,孢子重量与孢子计数之间的关系发生了变化。