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DNA fingerprinting differentiation between beta-carotene hyperproducer strains of Dunaliella from around the world.

Olmos J, Ochoa L, Paniagua-Michel J, Contreras R - Saline Syst. (2009)

Bottom Line: In this work, we applied our intron-sizing method to compare the 18S rDNA fingerprint between D. salina (CCAP 19/18), D. salina/bardawil (UTEX LB2538) and beta-carotene hyperproducing strains of Dunaliella isolated from salt saturated lagoons in Baja, Mexico.In Baja Mexico we found D. salina and D. salina/bardawil species by using intron-sizing-method.The National Center for Biotechnology Information (NCBI) Dunaliella 18S rDNA gene sequences were analyzed with our methodology and extraordinary correlation was found with experimental results.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Microbiology Laboratory, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Department of Marine Biotechnology, Ensenada, B.C, México.

ABSTRACT

Background: Dunaliella salina is the most important species of the genus for beta-carotene production. Several investigations have demonstrated that D. salina produces more than 10% dry weight of pigment and that the species grows in salt saturated lagoons. High plasticity in the green stage and the almost indistinguishable differences in the red phase make identification and differentiation of species and ecotypes very difficult and time consuming.

Results: In this work, we applied our intron-sizing method to compare the 18S rDNA fingerprint between D. salina (CCAP 19/18), D. salina/bardawil (UTEX LB2538) and beta-carotene hyperproducing strains of Dunaliella isolated from salt saturated lagoons in Baja, Mexico. All hyperproducer strains reached beta-carotene levels of about 10 pg/cell. Optical microscopy did not allow to differentiate between these Dunaliella strains; however, 18S rDNA fingerprinting methodology allowed us to differentiate D. salina from D. salina/bardawil.

Conclusion: In Baja Mexico we found D. salina and D. salina/bardawil species by using intron-sizing-method. The National Center for Biotechnology Information (NCBI) Dunaliella 18S rDNA gene sequences were analyzed with our methodology and extraordinary correlation was found with experimental results.

No MeSH data available.


Amplification with conserved and specific oligonucleotides. Lane 1, molecular weight marker. Lane 2 corresponds to amplification with MA1–MA2 conserved primers, using 19/18 DNA sample. Lane 3 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene hyperproducer species. Lane 4 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene non hyperproducer species.
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Figure 2: Amplification with conserved and specific oligonucleotides. Lane 1, molecular weight marker. Lane 2 corresponds to amplification with MA1–MA2 conserved primers, using 19/18 DNA sample. Lane 3 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene hyperproducer species. Lane 4 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene non hyperproducer species.

Mentions: Microscopic examination of Dunaliella salina 19/18 sample showed two different species of Dunaliella; one red and one green. Molecular fingerprinting determination using MA1–MA2 18S rDNA conserved primers from 19/18 DNA sample, gave two PCR products: one band of ~2100 bp that belongs to β-carotene hyperproducer species of D. salina, and a second product of ~1700 bp from Dunaliella species that never turns red (Fig. 2). These results agree with microscopic determination where two species of Dunaliella were observed (data not shown). Furthermore, using 19/18 DNA sample and DSs-MA2 specific primers we obtained a PCR product of ~700 bp from red species and no band from green species (Fig. 2). With these results we established the fingerprinting profile of 19/18 β-carotene hyperproducer strain of D. salina as (MA1–MA2 = ~2100 bp/DSs-MA2 = ~700 bp). Interestingly, the same fingerprinting was shown by D. salina M84320 isolated from Chile and reported by Wilcox and coworkers [12]. Furthermore, D. salina BCO2 strain isolated in Mexico, also presented the same 18S rDNA fingerprint [5,13] as did D. salina strain found in India by Raja and coworkers [14]. A common characteristic from these D. salina strains in addition to their fingerprinting profile was their β-carotene hyperproduction capacity.


DNA fingerprinting differentiation between beta-carotene hyperproducer strains of Dunaliella from around the world.

Olmos J, Ochoa L, Paniagua-Michel J, Contreras R - Saline Syst. (2009)

Amplification with conserved and specific oligonucleotides. Lane 1, molecular weight marker. Lane 2 corresponds to amplification with MA1–MA2 conserved primers, using 19/18 DNA sample. Lane 3 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene hyperproducer species. Lane 4 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene non hyperproducer species.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2710335&req=5

Figure 2: Amplification with conserved and specific oligonucleotides. Lane 1, molecular weight marker. Lane 2 corresponds to amplification with MA1–MA2 conserved primers, using 19/18 DNA sample. Lane 3 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene hyperproducer species. Lane 4 corresponds to amplification with DSs-MA2 specific-conserved oligonucleotides, from β-carotene non hyperproducer species.
Mentions: Microscopic examination of Dunaliella salina 19/18 sample showed two different species of Dunaliella; one red and one green. Molecular fingerprinting determination using MA1–MA2 18S rDNA conserved primers from 19/18 DNA sample, gave two PCR products: one band of ~2100 bp that belongs to β-carotene hyperproducer species of D. salina, and a second product of ~1700 bp from Dunaliella species that never turns red (Fig. 2). These results agree with microscopic determination where two species of Dunaliella were observed (data not shown). Furthermore, using 19/18 DNA sample and DSs-MA2 specific primers we obtained a PCR product of ~700 bp from red species and no band from green species (Fig. 2). With these results we established the fingerprinting profile of 19/18 β-carotene hyperproducer strain of D. salina as (MA1–MA2 = ~2100 bp/DSs-MA2 = ~700 bp). Interestingly, the same fingerprinting was shown by D. salina M84320 isolated from Chile and reported by Wilcox and coworkers [12]. Furthermore, D. salina BCO2 strain isolated in Mexico, also presented the same 18S rDNA fingerprint [5,13] as did D. salina strain found in India by Raja and coworkers [14]. A common characteristic from these D. salina strains in addition to their fingerprinting profile was their β-carotene hyperproduction capacity.

Bottom Line: In this work, we applied our intron-sizing method to compare the 18S rDNA fingerprint between D. salina (CCAP 19/18), D. salina/bardawil (UTEX LB2538) and beta-carotene hyperproducing strains of Dunaliella isolated from salt saturated lagoons in Baja, Mexico.In Baja Mexico we found D. salina and D. salina/bardawil species by using intron-sizing-method.The National Center for Biotechnology Information (NCBI) Dunaliella 18S rDNA gene sequences were analyzed with our methodology and extraordinary correlation was found with experimental results.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Microbiology Laboratory, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Department of Marine Biotechnology, Ensenada, B.C, México.

ABSTRACT

Background: Dunaliella salina is the most important species of the genus for beta-carotene production. Several investigations have demonstrated that D. salina produces more than 10% dry weight of pigment and that the species grows in salt saturated lagoons. High plasticity in the green stage and the almost indistinguishable differences in the red phase make identification and differentiation of species and ecotypes very difficult and time consuming.

Results: In this work, we applied our intron-sizing method to compare the 18S rDNA fingerprint between D. salina (CCAP 19/18), D. salina/bardawil (UTEX LB2538) and beta-carotene hyperproducing strains of Dunaliella isolated from salt saturated lagoons in Baja, Mexico. All hyperproducer strains reached beta-carotene levels of about 10 pg/cell. Optical microscopy did not allow to differentiate between these Dunaliella strains; however, 18S rDNA fingerprinting methodology allowed us to differentiate D. salina from D. salina/bardawil.

Conclusion: In Baja Mexico we found D. salina and D. salina/bardawil species by using intron-sizing-method. The National Center for Biotechnology Information (NCBI) Dunaliella 18S rDNA gene sequences were analyzed with our methodology and extraordinary correlation was found with experimental results.

No MeSH data available.