Isolation, Molecular Identification, Phototrophic, Mixotrophic and Heterotrophic Production of Chlorella sorokiniana

Öz

Microalgae was isolated using dilution and pouring plate method from the sea water sample taken from the Gumuldur region of Izmir. The species was identified as Chlorella sp. through morphologic evaluation under light microscope. Microalgae DNA was isolated through molecular methods and 16S and 18S rRNA gene regions were amplified by means of PCR. As a result of sequencing and phylogenetic evaluation of the gene regions, it was determined that the microalgae was Chlorella sorokiniana. Different methods such as centrifuge washing, antibiotic treatment, growth on agar and single cell isolation used to obtain axenic C. sorokiniana yielded successful results. Biomass and specific growth rate of phototrophic C. sorokiniana (0.19 g L-1 and 0.78 days-1, respectively), mixotrophic C. sorokiniana (0.31 g L-1 and 1.3 days-1, respectively) and heterotrophic C. sorokiniana (0.6 g L-1 and 2.52 days-1, respectively) were determined. According to the findings obtained, the biomass productivity of axenic
C. sorokiniana under different culture conditions can be ordered as: heterotrophy > mixotrophy > phototrophy.

Anahtar Kelimeler

• Chlorella sorokiniana
• molecular identification
• phototropy
• mixotrophy
• heterotrophy

Chlorella sorokiniana’ nın İzolasyonu, Moleküler Tanılanması, Fototrofik, Miksotrofik ve Heterotrofik Üretimi

Öz

İzmir’in Gümüldür bölgesinden alınan su örneğinde, seyreltme ve dökme plaka yöntemleri kullanılarak mikroalg izolasyonu yapılmıştır. Işık mikroskobuyla morfolojik olarak değerlendirilen türün Chlorella sp. olduğu saptanmıştır. Moleküler yöntemlerle mikroalg DNA’sı izole edilerek 16S ve 18S rRNA gen bölgeleri PCR’da çoğaltılmıştır. Bu dizinin sekanslanması ve filogenetik olarak değerlendirilmesi sonucu Chlorella sorokiniana olduğu belirlenmiştir. Aksenik C. sorokiniana elde etmek için santrifüj ile yıkama, antibiyotik ile muamele, agar ortamında büyütme ve tek hücre izolasyonu gibi farklı yöntemler kullanılarak aksenikleştirme işleminden başarılı sonuçlar elde edilmiştir. Fototrofik C. sorokiniana’ dan elde edilen biyokütle (0,19 g L-1) ve spesifik büyüme hızı (0,78 gün-1), miksotrofik C. sorokiniana’ dan elde edilen biyokütle (0,31 g L-1) ve spesifik büyüme hızı(1,3 gün-1), heterotrofik C. sorokiniana’ dan elde edilen biyokütle (0,6 g L-1) ve spesifik büyüme hızı (2,52 gün-1) olarak belirlenmiştir. Elde edilen bulgulara göre aksenik mikroalg C. sorokiniana’ nın farklı üretim koşullarındaki biyokütle verimliliği şu şeklide sıralanabilir: heterotrofi>miksotrofi>fototrofi.

Anahtar Kelimeler

• Chlorella sorokiniana
• moleküler tanılama
• Fototrofi
• Miksotrofi
• Heterotrofi

Kaynakça

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