In vitro regeneration, biological activities and phenolic composition of Astragalus gymnolobus Fischer, an endemic plant
Küçük Resim Yok
Tarih
2015
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Bolu Abant İzzet Baysal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Astragalus, Fabaceae ailesine ait yaklaşık 3000 türü kapsayan geniş bir cinstir. Astragalus türleri halk arasında antibakteriyel, antiviral, antifungal, antioksidant, antikanser, karaciğeri koruyucu, yara iyileştirici, antidiyabetik ve idrar söktürücü olarak kullanılmaktadır. Astragalus gymnolobus Fischer (gevenotu) endemik, bodur, yastık formunda çalılardır. A. gymnolobus için güvenilir bir bitki rejenerasyonu protokolü geliştirilmiştir. İki farklı eksplant (yaprak ve yaprak sapı) çeşitli bitki büyüme düzenleyicisi kombinasyonlarının bulunduğu MS (Murashige and Skoog) ortamı üzerinde kültüre alınmıştır.Yaprak eksplantları, yaprak sapı eksplantlarından daha fazla sürgün oluşturmuştur. En iyi sürgün oluşumu 0.5 mg/l TDZ (thidiazuron) ile yaprak eksplantlarından elde edilmiştir. En iyi kallus oluşumu 3.0 mg/l BA (benziladenin) ve 0.5mg/l 2,4-D (2,4-diklorofenoksiasetik asit) ile yaprak eksplantlarından elde edilmiştir. Rejenere olmuş sürgünler, farklı konsantrasyonlarda IAA (indol-3-asetik asit), IBA (indol-3-bütrik asit), 2.4-D veya NAA (naftalen asetik asit) içeren ortamlara konulmuştur. Ancak, rejenere olmuş sürgünler ile köklenme gözlemlenmemiştir. Toplanmış kalluslar A. gymnolobus bitkisinin biyolojik aktivite çalışmaları için kullanılmıştır. A. gymnolobus bitkisinin in-vitro yetiştirilmiş kallus ve doğada yetişen yaprak parçaları ile antibakteriyel ve antioksidant aktivite, toplam fenol ve flavonoid içeriği ve fenolik bileşenlerin HPLC (yüksek performanslı sıvı kromatografisi) analizi değerlendirilmiştir. A. gymnolobus bitkisinin su ve methanol özütlerinin antibakteriyel aktivitesi disk difüzyon metodu (Kirby-Bauer) ile belirlenmiştir. Genellikle, doğada yetişen yapraklar, in-vitro yetiştirilen kalluslardan daha iyi antibakteriyel aktivite göstermiştir. A. gymnolobus bitkisinin metanol özütlerinin serbest radikal süpürme gücü DPPH (2,2-difenil-1-pikrilhidrazil) metodu ile belirlenmiştir. En iyi antioksidant aktivite 25 ?g/ml konsantrasyonunda (91.47 %) doğada yetişen yaprakların metanol özütlerinde gözlemlenmiştir. Folin-Ciocaltaeu methodu kullanılarak toplam fenol içeriği ve alüminyum klorid (AlCl3) kolorimetrik methodu kullanılarak toplam flavonoid içeriği belirlenmiştir. Doğada yetişen yaprakların metanol özütleri, in-vitro yetiştirilen kalluslardan daha fazla fenol ve flavonoid içermektedir. Doğada yetişen yaprakların fenolik bileşenlerinin konsantrasyonu HPLC analizi ile in-vitro yetiştirilen kalluslardan yüksek bulunmuştur.
Astragalus is a large genus covering about 3000 species belonging to Fabaceae family. Astragalus species have been used as antibacterial, antiviral, antifungal, antioxidant, anticancer, hepatoprotective, wound healing, anti-diabetic and diuretic in folk medicine. Astragalus gymnolobus Fischer (milkvetch weed) is an endemic dwarf cushion-forming shrub. An efficient plant regeneration system was developed for A. gymnolobus. Two different explants (leaf and petiole) were cultured on MS (Murashige and Skoog) medium with various plant growth regulator combinations. Leaf explants formed more shoots than petiole explants. The best shoot formation was obtained from leaf explants with 0.5 mg/l TDZ (thidiazuron). The best callus formation was obtained with 3.0 mg/l BA (benzyladenine) and 0.5 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) with leaf explant. Regenerated shoots were put on medium containing different concentrations of IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), 2,4-D or NAA (naphthalene acetic acid). However, rooting was not observed with regenerated shoots. Collected calluses were used for biological activity studies of A. gymnolobus. Antibacterial and antioxidant activities, total phenol and flavonoid content, and HPLC (high performance liquid chromatograph) analysis of phenolic constituents of field-grown leaves and in vitro-grown callus parts of A. gymnolobus were evaluated. Antibacterial activity of water and methanol extracts of A. gymnolobus was determined with Kirby-Bauer disk diffusion assay. Generally, field-grown leaves showed better antibacterial activities than in vitro-grown callus. The free radical scavenging potency of methanol extracts of A. gymnolobus was determined with DPPH (2,2-diphenyl-1-picrylhydrazil) method. The best antioxidant activity was obtained with methanol extract of field-grown leaves at 25 ?g/ml concentrations (91.47 %). Total phenolic content by using Folin-Ciocaltaeu method and the total flavonoid content by using aluminum chloride (AlCl3) colorimetric method were determined. Methanol extract of field-grown leaves contained more phenols and flavonoid than in vitro-grown callus. Concentrations of phenolic compounds in field-grown leaves were found higher than the in vitro-grown callus with HPLC analysis.
Astragalus is a large genus covering about 3000 species belonging to Fabaceae family. Astragalus species have been used as antibacterial, antiviral, antifungal, antioxidant, anticancer, hepatoprotective, wound healing, anti-diabetic and diuretic in folk medicine. Astragalus gymnolobus Fischer (milkvetch weed) is an endemic dwarf cushion-forming shrub. An efficient plant regeneration system was developed for A. gymnolobus. Two different explants (leaf and petiole) were cultured on MS (Murashige and Skoog) medium with various plant growth regulator combinations. Leaf explants formed more shoots than petiole explants. The best shoot formation was obtained from leaf explants with 0.5 mg/l TDZ (thidiazuron). The best callus formation was obtained with 3.0 mg/l BA (benzyladenine) and 0.5 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) with leaf explant. Regenerated shoots were put on medium containing different concentrations of IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), 2,4-D or NAA (naphthalene acetic acid). However, rooting was not observed with regenerated shoots. Collected calluses were used for biological activity studies of A. gymnolobus. Antibacterial and antioxidant activities, total phenol and flavonoid content, and HPLC (high performance liquid chromatograph) analysis of phenolic constituents of field-grown leaves and in vitro-grown callus parts of A. gymnolobus were evaluated. Antibacterial activity of water and methanol extracts of A. gymnolobus was determined with Kirby-Bauer disk diffusion assay. Generally, field-grown leaves showed better antibacterial activities than in vitro-grown callus. The free radical scavenging potency of methanol extracts of A. gymnolobus was determined with DPPH (2,2-diphenyl-1-picrylhydrazil) method. The best antioxidant activity was obtained with methanol extract of field-grown leaves at 25 ?g/ml concentrations (91.47 %). Total phenolic content by using Folin-Ciocaltaeu method and the total flavonoid content by using aluminum chloride (AlCl3) colorimetric method were determined. Methanol extract of field-grown leaves contained more phenols and flavonoid than in vitro-grown callus. Concentrations of phenolic compounds in field-grown leaves were found higher than the in vitro-grown callus with HPLC analysis.
Açıklama
Fen Bilimleri Enstitüsü, Biyoloji Ana Bilim Dalı
Anahtar Kelimeler
Biyoloji, Biology ; Biyoteknoloji
