Lipid droplets of bacteria, algae and fungi and a relationship between their contents and genome sizes as revealed by BODIPY and DAPI staining

Tsuneyoshi Kuroiwa, Mio Ohnuma, Yuuta Imoto, Osami Misumi, Takayuki Fujiwara, Shin Ya Miyagishima, Nobuko Sumiya, Haruko Kuroiwa

研究成果: Article

8 引用 (Scopus)

抄録

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 nonoleaginous. 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.

元の言語English
ページ(範囲)289-299
ページ数11
ジャーナルCytologia
77
発行部数3
DOI
出版物ステータスPublished - 2012 9
外部発表Yes

Fingerprint

algae
Genome Size
droplets
Fungi
Staining and Labeling
Bacteria
fungi
genome
bacteria
lipids
Chlamydomonas reinhardtii
Chloroplasts
microscopes
staining
DAPI
Lipid Droplets
Anabaena flos-aquae
chloroplasts
Klebsormidium
Cyanidium caldarium

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Plant Science
  • Animal Science and Zoology

これを引用

Lipid droplets of bacteria, algae and fungi and a relationship between their contents and genome sizes as revealed by BODIPY and DAPI staining. / Kuroiwa, Tsuneyoshi; Ohnuma, Mio; Imoto, Yuuta; Misumi, Osami; Fujiwara, Takayuki; Miyagishima, Shin Ya; Sumiya, Nobuko; Kuroiwa, Haruko.

:: Cytologia, 巻 77, 番号 3, 09.2012, p. 289-299.

研究成果: Article

Kuroiwa, Tsuneyoshi ; Ohnuma, Mio ; Imoto, Yuuta ; Misumi, Osami ; Fujiwara, Takayuki ; Miyagishima, Shin Ya ; Sumiya, Nobuko ; Kuroiwa, Haruko. / Lipid droplets of bacteria, algae and fungi and a relationship between their contents and genome sizes as revealed by BODIPY and DAPI staining. :: Cytologia. 2012 ; 巻 77, 番号 3. pp. 289-299.
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abstract = "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 nonoleaginous. 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.",
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T1 - Lipid droplets of bacteria, algae and fungi and a relationship between their contents and genome sizes as revealed by BODIPY and DAPI staining

AU - Kuroiwa, Tsuneyoshi

AU - Ohnuma, Mio

AU - Imoto, Yuuta

AU - Misumi, Osami

AU - Fujiwara, Takayuki

AU - Miyagishima, Shin Ya

AU - Sumiya, Nobuko

AU - Kuroiwa, Haruko

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AB - 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 nonoleaginous. 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.

KW - Algae

KW - BODIPY

KW - Cyanidioschyzon merolae

KW - Fungi

KW - Genome size

KW - Lipid droplets

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