The Effects of Silver Nitrate and Pre-Cold Treatments on Callus Formation in Strawberry Anther Culture

Year 2024, Volume: 21 Issue: 1, 15 - 23, 30.01.2024

Sevinç Şener , Ayşe Gül Nasırcılar , Ahmet Karaçan

https://doi.org/10.33462/jotaf.1217960

Abstract

Strawberry (Fragaria x ananassa Duch.) which is one of the widely grown berry species in the world has economic and commercial importance. In commercial strawberry varieties, in order to increase yield and quality, it is necessary to obtain starting materials that are resistant/tolerant to biotic and abiotic stress factors. Biotechnological methods have an important place in strawberry breeding studies due to the long and costly process of classical breeding methods, the genetic expansion of seed production, high ploidy level and strong heterozygosity. Haploid plant production is an efficient breeding method that has been successfully applied to most plant species. However, due to the lack of sufficient haploid studies on strawberry and the fact that a specific protocol for this species has not yet been developed the necessary progress has not been made in this regard. In this study, the effectiveness of some factors determine the success in anther culture which has a significant place in obtaining haploid strawberries was investigated. For this reason, first, different sodium hypochlorite doses (NaOCl; 1%, 2%, 3%) and application durations (10, 15, 20 min) were used to determine the appropriate method for sterilisation, then cold pre-treatments (24, 36, 48, 72 hours at +4 °C) and different silver nitrate doses (AgNO3; 10, 20, 30, 40 mg l-1) were employed for callus induction in Festival strawberry variety. At the conclusion of the study it was observed that the lowest contamination rate (1%) was obtained by soaking in 1% sodium hypochlorite solution for 10 minutes. Cold pre-treatment of flower buds at +4 °C for 36 hours produced the highest callus induction rate (96%). The evaluation of the effect of AgNO3 application at different doses on the callus induction rate revealed that the highest callus induction (82%) was obtained from 20 mg l-1 AgNO3 dosage. This study showed that anther culture practices in strawberry can be improved by using cold pre-treatment, appropriate sterilization method and silver nitrate addition to the medium.

Keywords

Anther, Callogenesis, Silver nitrate, Micropropagation, Haploid

Gümüş Nitrat ve Ön Soğuk Uygulamalarının Çilek Anter Kültüründe Kallus Oluşumuna Etkileri

Abstract

Çilek (Fragaria x ananassa Duch.), dünyada yaygın olarak yetiştiriciliği yapılan, ekonomik ve ticari öneme sahip olan önemli üzümsü meyve türlerinden birisidir. Ticari çilek çeşitlerinde, verim ve kalitenin artırılması için biotik ve abiotik stress faktörlerine karşı dayanıklı veya tolerant başlangıç materyallerinin elde edilmesi gerekmektedir. Çilekte, klasik ıslah çalışmaları süreçlerinin uzun ve masraflı olması, tohumdan üretiminin genetik açılım göstermesi, yüksek ploidi seviyesi ve güçlü heterozigotluk sebebiyle, biyoteknolojik yöntemler ıslah çalışmalarında önemli bir yer tutmaktadır. Haploid bitki üretimi çoğu bitki türünde başarı ile uygulanan etkili bir ıslah yöntemidir. Ancak çilekte haploid bitki üretimi ile ilgili yeterli sayıda çalışmanın olmaması ve bu türe özgü bir protokolün hala geliştirilememesi gibi nedenlerle bu konuda gerekli ilerlemeler sağlanamamıştır. Bu çalışmada, haploid çilek eldesinde önemli bir yer tutan anter kültüründe başarıyı etkileyen bazı faktörlerin etkinliğinin araştırılması amaçlanmıştır. Bu nedenle, Festival çilek çeşidinde kallus indüksiyonu için öncelikle sterilizasyon için uygun yöntemi belirlemek amacıyla farklı sodyum hipoklorit dozları (NaOCl; %1, %2, %3) ve uygulama süreleri (10, 15, 20 dk) ardından farklı soğuk ön uygulamaları (+4 oC'de 24, 36, 48, 72 saat) ve gümüş nitrat dozları (AgNO3; 10, 20, 30, 40 mg l-1) kullanılmıştır. Araştırmadan elde edilen verilerin değerlendirilmesi sonucunda en düşük kontaminasyon oranının (%1) %1'lik sodyum hipoklorit çözeltisinde 10 dakika bekletilerek elde edildiği belirlenmiştir. Çiçek tomurcuklarının 36 saat boyunca +4 oC 'de soğuk ön muameleye tabi tutulması ile en yüksek kallus indüksiyon oranı (%96) sağlanmıştır. Farklı dozlarda AgNO3 uygulamasının kallus indüksiyonu üzerine etkisinin değerlendirilmesi ise en yüksek oranın (%82) 20 mg l-1 AgNO3 dozundan elde edildiğini göstermiştir. Bu çalışma, çilekte anter kültürü çalışmalarının soğuk ön uygulaması, uygun sterilizasyon yöntemi ve besi ortamına gümüş nitrat ilavesi ile geliştirilebileceğini göstermiştir.

Keywords

Anter, Kallogenesis, Gümüş nitrat, Mikroçoğaltım, Haploid

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