The Influence of Genotype, Pre-Chilling and Nutrient Media on Callus Induction and Plantlet Regeneration in Rye (Secale cereale L.) Anther Culture

Year 2019, Issue: 16, 559 - 566, 31.08.2019

Nüket Altındal , İlknur Akgün

https://doi.org/10.31590/ejosat.581763

Abstract

The present study investigated the effects of genotypes (diploid and tetraploid), of pre-treatment (coldtreatmentwithin one week at 4°C and non-cold treatment) and and nutrient media (MS, N6 and NN) on anther response ratio,callus induction and green plantlet regeneration in anther culture of rye. In our experiment, the MSmedium (consisting of 10 g/L of agar), NN medium (consisting of 8 g/L agar) and N6 medium (consisting of 5 g/L of agar)were used for callus formation, MS and NN media supplemented with 2.4 µmol/L of KIN, 4.4 µmol/L of BAP and 2.7 µmol/L of NAA as hormone and 30.3 g/L of sucrose as carbon source. Additionaly, N6 medium supplemented with 2 mg/L 2.4-D, 0.5 mg/L KIN, 1 mg/L IAA as hormone and 90 g/L sucrose as carbon source. Two media (MS and NN) used for callus formation were the same for plantlet regeneration, however N6 media was modified and included 2 mg/L KIN, 2 g/L activated charcoal, 20 g/L sucrose and 5 g/L agar. Our study demonstrated the negativeeffect of pre-chilling on anther response ratio and callus size, but it also showed that the positive effect on callus ratio and root formation.On the other hand, effect of pre-chilling on investigated characters changed according to rye genotypes and was significant (except for anther response ratio). While effect of nutrient media on callus ratio and size was significant, it was not significant on anther response ratio. The highest root and plantlet regeneration ratio were obtained as a result of cultured on MS media of pre-chilled anthers in diploid rye.

Keywords

Anther culture, in vitro, callus, rye

Çavdarda (Secale cereale L.) Anter Kültürü ile Kallus Oluşumu ve Bitki Rejenerasyonu Üzerine Genotip, Soğuk Uygulama ve Besi Ortamlarının Etkisi

Abstract

Bu çalışmada, çavdarda genotipin (diploid
ve tetraploid), ön işlem (+4 ^oC’de 1 hafta süre ön işlem uygulanmış
ve uygulanmamış) ve besin ortamlarının (MS, N₆ ve NN) anter kültüründe
anter tepki oranı, kallus oluşumu ve bitki rejenerasyonu üzerine etkileri
incelenmiştir. Denemede kallus oluşum ortamı olarak MS ortamında 10 g/L agar,
NN ortamında 8 g/L agar kullanılmış; 2,4 µmol/L Kinetin, 4,4 µmol/L BAP, 2,7
µmol/L NAA ve karbon kaynağı olarak 30,3 g/L sükroz her iki ortama ilave
edilmiştir. N₆ ortamına ise 5 g/L agar, 2 mg/L 2,4-D, 0,5 mg/L
Kinetin, 1 mg/L IAA ve 90 g/L sükroz ilave edilmiştir. Rejenerasyon ortamı
olarak modifiye edilmiş N₆ ortamına 2 g/L aktif karbon, karbon
kaynağı olarak 20 g/L sükroz, hormon olarak 2 mg/L kinetin, katılaştırıcı madde
olarak da 5 g/L agar eklenmiştir. Araştırmada soğuk ön işlem uygulamasının
anter tepki oranı ve kallus büyüklüğü üzerine olumsuz, kallus oranı ve bitki
rejenerasyonu üzerine olumlu etkisi belirlenmiştir. Diğer taraftan soğuk ön
uygulamasının incelenen özellikler üzerine etkisi çavdar genotiplerinde
farklılık göstermiştir. Ayrıca anter tepki oranı hariç, incelenen diğer özellikler
üzerine ön işlem uygulamasının etkisi önemli, genotipin etkisi ise önemsiz
çıkmıştır. Besin ortamlarının etkisi ise kallus oranı ve büyüklüğü üzerinde
önemli, anter tepki oranı üzerine ise önemsiz bulunmuştur. En fazla kök oluşumu
ve bitki rejenerasyonu soğuk uygulamalı diploid çavdar anterlerinin MS
ortamında kültüre alınması ile elde edilmiştir.

Keywords

Anter kültürü, in vitro, kallus

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