Lipid Droplets of Bacteria, Algae and Fungi and a Relationship between their Contents and Genome Sizes as Revealed by BODIPY and DAPI Staining
Lipid Droplets of Bacteria, Algae and Fungi and a Relationship between their Contents and Genome Sizes as Revealed by BODIPY and DAPI Staining
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DOI：
10.1508/cytologia.77.289
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发表时间：
2012-09-01
期刊：
影响因子：
1
通讯作者：
Kuroiwa, Haruko
中科院分区：
文献类型：
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作者：
Kuroiwa, Tsuneyoshi;Ohnuma, Mio;Kuroiwa, Haruko
我们开发了一种简便的方法，通过用二氟化硼(BODIPY)染色来显示某些细菌、藻类和真菌中充满三酰甘油的脂滴(LDS)。当BODIPY被蓝光激发时，LDS发出绿色荧光，这与叶绿体的红色自体荧光很容易区分。这使得BODIPY染色适合于少量LDS的鉴定，特别是在植物中。我们首先确保了在氮充足(+N)和缺氮(-N)的培养基中生长的莱茵衣藻细胞中，BODIPY染色的LDS斑点与尼罗红染色的LDS斑点一致。此外，用视频增强显微镜光计数系统(VIMPCS)显示，N饥饿细胞的每个细胞的LD含量是对照(+N)的200倍。用BODIPY染色显示细菌、藻类和真菌中的LD，包括那些被认为不含油性的藻类。我们在单细胞和多细胞细菌和真核生物中发现了LD斑点，即斑点蓝藻、三角蓝藻、莱茵衣藻、尼氏克雷伯氏菌和青霉，但在水华鱼腥藻中没有。我们还利用VIMPCS研究了藻类和真菌中LDS含量与基因组大小的关系，但未能发现基因组大小与LDS产量之间的强烈关系。最后，我们认为LDS的位置与细胞器有关，包括内质网和叶绿体，它们与LDS的形成有关。
We have developed a convenient method to visualize triacylglycerol-filled lipid droplets (LDs) in some species of bacteria, algae and fungi by staining with borondipyrromethene difluoride (BODIPY). When BODIPY was excited by blue light, LDs emitted green fluorescence, which was distinguished easily from the red autofluorescence of chloroplasts. This makes BODIPY staining suitable for the identification of small amounts of LDs, especially in plants. We first ensured that in Chlamydomonas reinhardtii cells growing in nitrogen-replete (+N) and -deficient (-N) media, the spots of BODIPY-stained LDs coincided with those of Nile Red-stained LDs. In addition, it was shown that the LD content per cell in N-starved cells was 200-fold higher than those of the control (+N) using a video-intensified microscope photoncounting system (VIMPCS). BODIPY staining was applied to visualize LD in bacteria, algae and fungi, and included those algae regarded as non-oleaginous. We identified LD spots in unicellular and multicellular bacteria and eukaryotes, namely Cyanidioschyzon merolae, Cyanidium caldarium delta, Chlamydomonas reinhardtii, Klebsormidium nitens and Penicillium sp., but not in Anabaena flos-aquae. We also examined the relationship between the contents of LDs and the genome size in the algae and fungi using VIMPCS but were unable to find a strong relationship between genome size and production of LDs. Finally, the location of LDs was considered in relation to organelles including the endoplasmic reticulum and chloroplasts, which are related to the formation of LDs.