15. The incubation of several colonies in the same Petri dish is
recommended because of the slower radial growth of
F. fujikuroi compared to other fungi.
16. The fluorescent tubes should not be too close to the cultures to
avoid excessive heating.
17. Use two 2-ml screw-cap microfuge tubes for the same sample
in order to have two replicates.
18. Fill each screw-cap microtube with approximately 100 μl
of sand.
19. The recommended amount of dry mycelium depends on its
carotenoid content. Approximately 20 mg of dry mycelium are
sufficient for samples from strains overproducing carotenoids,
and a higher weight (at least 40–50 mg) are needed for the
samples with lower carotenoid content, as those of the wild
type in the dark.
20. Because of the presence of polar carotenoids in Fusarium,
acetone is an appropriate solvent for extraction of carotenoids.
21. All manipulations during carotenoid extraction and measurement should be done under green or red light to prevent
isomerization and photodegradation of carotenoids.
22. It is important that the supernatant to be dried in the speed
vacuum concentrator is totally clean.
23. Do not fill 2-ml microtubes above 1 ml because the sample
could spill in the speed vacuum concentrator.
24. If a sample has a very low of carotenoid content, dissolve the
sample in only 100 μl of acetone. This requires the availability
of microcells for spectrophotometer measuring.
25. If carotenoids were not going to be analyzed immediately after
the extraction, extracts should be stored at À20
C (or lower)
until spectrophotometer analysis in order to prevent degradation of carotenoids.
26. If maximal absorbance of the spectrum exceeds the value of
1, dilute the carotenoid extract accordingly to get a lower
absorbance.
27. Carotenoids of F. fujikuroi usually have a maximal absorption
peak between 470 and 480 nm.
Acknowledgments
We thank the Spanish Government (projects BIO2012-39716,
BIO2015-69613-R, and CaRed Networks BIO2015-71703REDT and BIO2017-90877-REDT), and Junta de Andalucı ´a
(project CTS-6638) for financial support. J.P-M was granted by
358
Julia Marente et al.
recommended because of the slower radial growth of
F. fujikuroi compared to other fungi.
16. The fluorescent tubes should not be too close to the cultures to
avoid excessive heating.
17. Use two 2-ml screw-cap microfuge tubes for the same sample
in order to have two replicates.
18. Fill each screw-cap microtube with approximately 100 μl
of sand.
19. The recommended amount of dry mycelium depends on its
carotenoid content. Approximately 20 mg of dry mycelium are
sufficient for samples from strains overproducing carotenoids,
and a higher weight (at least 40–50 mg) are needed for the
samples with lower carotenoid content, as those of the wild
type in the dark.
20. Because of the presence of polar carotenoids in Fusarium,
acetone is an appropriate solvent for extraction of carotenoids.
21. All manipulations during carotenoid extraction and measurement should be done under green or red light to prevent
isomerization and photodegradation of carotenoids.
22. It is important that the supernatant to be dried in the speed
vacuum concentrator is totally clean.
23. Do not fill 2-ml microtubes above 1 ml because the sample
could spill in the speed vacuum concentrator.
24. If a sample has a very low of carotenoid content, dissolve the
sample in only 100 μl of acetone. This requires the availability
of microcells for spectrophotometer measuring.
25. If carotenoids were not going to be analyzed immediately after
the extraction, extracts should be stored at À20
C (or lower)
until spectrophotometer analysis in order to prevent degradation of carotenoids.
26. If maximal absorbance of the spectrum exceeds the value of
1, dilute the carotenoid extract accordingly to get a lower
absorbance.
27. Carotenoids of F. fujikuroi usually have a maximal absorption
peak between 470 and 480 nm.
Acknowledgments
We thank the Spanish Government (projects BIO2012-39716,
BIO2015-69613-R, and CaRed Networks BIO2015-71703REDT and BIO2017-90877-REDT), and Junta de Andalucı ´a
(project CTS-6638) for financial support. J.P-M was granted by
358
Julia Marente et al.