Department of Biological Sciences, University of Lethbridge, Lethbridge, T1K 3M4 Alberta, Canada.
Mutat Res. 2011 Feb 10;707(1-2):61-6. doi: 10.1016/j.mrfmmm.2010.12.013. Epub 2011 Jan 12.
In earlier studies, we showed that abiotic stresses, such as ionizing radiation, heavy metals, temperature and water, trigger an increase in homologous recombination frequency (HRF). We also demonstrated that many of these stresses led to inheritance of high-frequency homologous recombination, HRF. Although an increase in recombination frequency is an important indicator of genome rearrangements, it only represents a minor portion of possible stress-induced mutations. Here, we analyzed the influence of heat, cold, drought, flood and UVC abiotic stresses on two major types of mutations in the genome, point mutations and small deletions/insertions. We used two transgenic lines of Arabidopsis thaliana, one allowing an analysis of reversions in a stop codon-containing inactivated β-glucuronidase transgene and another one allowing an analysis of repeat stability in a microsatellite-interrupted β-glucuronidase transgene. The transgenic Arabidopsis line carrying the β-glucuronidase-based homologous recombination substrate was used as a positive control. We showed that the majority of stresses increased the frequency of point mutations, homologous recombination and microsatellite instability in somatic cells, with the frequency of homologous recombination being affected the most. The analysis of transgenerational changes showed an increase in HRF to be the most prominent effect observed in progeny. Significant changes in recombination frequency were observed upon exposure to all types of stress except drought, whereas changes in microsatellite instability were observed upon exposure to UVC, heat and cold. The frequency of point mutations in the progeny of stress-exposed plants was the least affected; an increase in mutation frequency was observed only in the progeny of plants exposed to UVC. We thus conclude that transgenerational changes in genome stability in response to stress primarily involve an increase in recombination frequency.
在早期的研究中,我们表明非生物胁迫,如电离辐射、重金属、温度和水,会引发同源重组频率(HRF)的增加。我们还证明,许多这些应激导致高频同源重组的遗传。虽然重组频率的增加是基因组重排的一个重要指标,但它只代表了可能的应激诱导突变的一小部分。在这里,我们分析了热、冷、干旱、洪水和 UVC 非生物胁迫对基因组中两种主要类型突变的影响,点突变和小的缺失/插入。我们使用了两个拟南芥的转基因系,一个用于分析含有失活β-葡萄糖醛酸酶基因的终止密码子的回复突变,另一个用于分析微卫星中断的β-葡萄糖醛酸酶基因的重复稳定性。携带基于β-葡萄糖醛酸酶的同源重组底物的转基因拟南芥系被用作阳性对照。我们表明,大多数应激增加了体细胞中点突变、同源重组和微卫星不稳定性的频率,其中同源重组的频率受影响最大。对跨代变化的分析表明,HRF 的增加是在后代中观察到的最显著的效应。除了干旱之外,所有类型的应激都导致重组频率的显著变化,而微卫星不稳定性的变化仅在暴露于 UVC、热和冷的植物后代中观察到。暴露于应激的植物后代中的点突变频率受影响最小;仅在暴露于 UVC 的植物后代中观察到突变频率的增加。因此,我们得出结论,对基因组稳定性的跨代应激反应主要涉及重组频率的增加。