Maligoy Mathieu, Mercade Myriam, Cocaign-Bousquet Muriel, Loubiere Pascal
INSA-LISBP, 135 Avenue de Rangueil, 31077 Toulouse cedex 4, France.
Appl Environ Microbiol. 2008 Jan;74(2):485-94. doi: 10.1128/AEM.01531-07. Epub 2007 Nov 9.
The study of microbial interactions in mixed cultures remains an important conceptual and methodological challenge for which transcriptome analysis could prove to be the essential method for improving our understanding. However, the use of whole-genome DNA chips is often restricted to the pure culture of the species for which the chips were designed. In this study, massive cross-hybridization was observed between the foreign cDNA and the specific Lactococcus lactis DNA chip. A very simple method is proposed to considerably decrease this nonspecific hybridization, consisting of adding the microbial partner's DNA. A correlation was established between the resulting cross-hybridization and the phylogenetic distance between the microbial partners. The response of L. lactis to the presence of Saccharomyces cerevisiae was analyzed during the exponential growth phase in fermentors under defined growth conditions. Although no differences between growth kinetics were observed for the pure and the mixed cultures of L. lactis, the mRNA levels of 158 genes were significantly modified. More particularly, a strong reorientation of pyrimidine metabolism was observed when L. lactis was grown in mixed cultures. These changes in transcript abundance were demonstrated to be regulated by the ethanol produced by the yeast and were confirmed by an independent method (quantitative reverse transcription-PCR).
对混合培养物中微生物相互作用的研究仍然是一个重要的概念和方法挑战,转录组分析可能被证明是增进我们理解的关键方法。然而,全基因组DNA芯片的使用通常仅限于为其设计芯片的物种的纯培养。在本研究中,观察到外源cDNA与特定的乳酸乳球菌DNA芯片之间存在大量交叉杂交。提出了一种非常简单的方法来显著减少这种非特异性杂交,即添加微生物伙伴的DNA。在所得交叉杂交与微生物伙伴之间的系统发育距离之间建立了相关性。在确定的生长条件下,在发酵罐的指数生长期分析了乳酸乳球菌对酿酒酵母存在的反应。尽管乳酸乳球菌的纯培养物和混合培养物在生长动力学上未观察到差异,但158个基因的mRNA水平发生了显著改变。更特别的是,当乳酸乳球菌在混合培养物中生长时,观察到嘧啶代谢有强烈的重新定向。这些转录本丰度的变化被证明受酵母产生的乙醇调节,并通过独立方法(定量逆转录PCR)得到证实。