Suzuki T, Hitomi A, Magee P T, Sakaguchi S
Department of Biology, Faculty of Science, Nara Women's University, Japan.
J Bacteriol. 1994 Jun;176(11):3345-53. doi: 10.1128/jb.176.11.3345-3353.1994.
In order to clarify the relationship between polyploidization and the capability of phenotypic switching in the imperfect yeast Candida albicans, two types of variants were isolated as segregants from a fusant, which produced a proportion of the cell population with a higher ploidy than the rest, either in a temperature-dependent or -independent manner, when incubated at low (28 degrees C) and high (37 degrees C) temperatures. In the case of the temperature-dependent type of variants, high-ploidy cells appeared at 37 degrees C but rarely at 28 degrees C. This phenotype was named Pldts (temperature-sensitive polyploidization), and the temperature-independent phenotype was called Pld-. The appearance of high-ploidy cells in the culture of the Pldts strain at 37 degrees C was accompanied by a significant increase in the frequency of auxotrophic variants; these variants probably occur as a result of segregation of auxotrophic markers from the heterozygous to the homozygous state. Both Pldts and Pld- phenotypes were recessive in a fusion with a Pld+ parent. An adenine auxotrophic marker (ade1) was introduced into a Pldts strain in a heterozygous state, and the individual high-ploidy cells of this strain, grown at 37 degrees C, were micromanipulated to form colonies, which consisted of red and white sectors appearing at high frequency on a pink background. When the ade1 auxotrophy was introduced into Pld- strains, frequently sectored colonies were produced. These results suggested an increased level of chromosome missegregation in both types of Pld mutants. Analyses by pulsed-field gel electrophoresis of Ade-segregants, derived from a micromanipulated high-ploidy cell of a Pld(ts) strain, suggested the occurrence of nonreciprocal recombination, some of which includes chromosome loss.
为了阐明多倍体化与不完全酵母白色念珠菌表型转换能力之间的关系,从一个融合体中分离出两种类型的变体作为分离子,当在低温(28℃)和高温(37℃)下培养时,该融合体产生一定比例的细胞群体,其倍性高于其余细胞群体,且这种现象以温度依赖性或非依赖性方式出现。对于温度依赖性变体类型,高倍体细胞在37℃时出现,但在28℃时很少出现。这种表型被命名为Pldts(温度敏感型多倍体化),非温度依赖性表型被称为Pld-。Pldts菌株在37℃培养时高倍体细胞的出现伴随着营养缺陷型变体频率的显著增加;这些变体可能是由于营养缺陷型标记从杂合状态分离到纯合状态而产生的。Pldts和Pld-表型在与Pld+亲本融合时都是隐性的。将腺嘌呤营养缺陷型标记(ade1)以杂合状态引入Pldts菌株,该菌株在37℃下生长的单个高倍体细胞经显微操作形成菌落,这些菌落由在粉红色背景上高频出现的红色和白色扇形区域组成。当将ade1营养缺陷型引入Pld-菌株时,会产生频繁出现扇形区域的菌落。这些结果表明两种类型的Pld突变体中染色体错分离水平增加。对源自Pld(ts)菌株显微操作的高倍体细胞的Ade分离子进行脉冲场凝胶电泳分析,表明发生了非相互重组,其中一些包括染色体丢失。