Priming edilmiş endüstriyel kenevir (Cannabis sativa L.) tohumlarının in vitro çimlenmesinin optimize edilmesi

Abstract

Endüstriyel kenevir (Cannabis sativa L., Cannabaceae) bitkisinin in vitro çimlenmesi, genom düzenleme gibi biyoteknolojik araçların uygulanması için oldukça önemlidir. Bu nedenle, tohumlar yüzey sterilize edildikten sonra 24, 48 ve 72 saat boyunca hidrojen peroksit (H2O2) ve steril distile su (dH2O) ile priming uygulama yapılmıştır. Daha sonra tohumlar, 1.0 mg/L BAP ve 200 mg/L antibiyotik ile zenginleştirilmiş Murashige ve Skoog (MS) ortamı üzerinde kültüre alınmıştır. Tohumların H2O2 ile yapılan priming sonucunda daha iyi sonuç verdiğini ve %100 çimlenme olduğu gözlemlenmiştir. Buna karşı hidropriming ile yapılan çalışmada 30-52,5 çimlenme gözlenirken, en iyi sonucun 48 saatlik priming sonucu olduğu kaydedilmiştir. Sürgün sayılarına bakıldığında en fazla 1,98 sürgün, %3,0 H2O2 ve 72 saatlik priming süresi kombinasyonundan elde edilmiştir. Sonuçlar ayrıca kutu grafiği, normal grafikler, kontur grafikleri ve yüzey grafikleri gibi farklı istatistiksel grafikler oluşturularak da analiz edilmiştir. Normal grafikler, H2O2 konsantrasyonunun her iki çıktı değişkenindeki önemini açıkça göstermiştir. Kontur ve yüzey grafikleri ise çıktı verilerini farklı alt gruplara ayırmış ve sonuçları doğrulanmıştır.

Keywords

• Çimlenme
• endüstriyel kenevir
• in vitro
• priming
• istatistiksel analiz

Optimizing in vitro germination of primed industrial hemp (Cannabis sativa L.) seeds

Abstract

Germination under in vitro conditions for industrial hemp (Cannabis sativa L., Cannabaceae) is highly significant for the application of biotechnological tools like genome editing. Therefore, seeds were surface sterilized followed by priming with hydrogen peroxide (H2O2) and sterile distilled water (dH2O) for 24, 48, and 72 h. The primed seeds were inoculated on the Murashige and Skoog (MS) medium enriched with 1.0 mg/L benzyl amino purine (BAP) and 200 mg/L antibiotics. Exposing seeds to H2O2 was superior and 100% germination was observed. Whereas hydropriming resulted in 30-52.5% germination with a maximum of 48h priming time. The results on shoot counts revealed a maximum of 1.98 shoots from the combination of 3.0% H2O2 and 72h priming time. The results were also analyzed by constructing different statistical plots like box plots, normal plots, contour plots, and surface plots. The normal plots exhibited the significance of H2O2 concentration on both output variables. Whereas contour and surface plots classified the output data into different sub-groups and confirmed the results.

Keywords

• Germination
• industrial hemp
• in vitro
• priming
• statistical analysis

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