Azot Protoksit ve Orizalinin ‘41 B’ ile ‘Fercal’ Asma Anaçlarında Mutajenik Etkileri
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Dosyalar
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Asma (Vitis spp.), küresel boyutta sosyal ve ekonomik açıdan en önemli bitkilerden birisidir.
Üzüm çeşitleri ve asma anaçlarının geliştirilmesi günümüzdeki tüketici taleplerinin yanı sıra iklim
değişikliğine uyum sağlamak için giderek artan bir öneme sahiptir. Bugüne kadar seleksiyon ve
melezleme çalışmalarıyla elde edilen asma anaçları, tüm talepleri karşılayamamaktadır. Çoğu asma anacı
Avrupa bağ bölgeleri için seçilmiş olsa da ülkemizdeki kireçli ve sığ topraklarda, kısa vejetasyonlu bağ
alanlarında üretici gereksinimlerini tam olarak karşılayamamaktadırlar. Asma ıslahında doğal genetik
çeşitlilikten yeni genotipler elde etmede mutantlar, melezlemeye kıyasla daha umut vericidir. Poliploit
bitkiler, diploit akrabalarına kıyasla çeşitli açılardan daha iyi performans gösterirler. Bu çalışmada, 41 B
ve Fercal asma anaçlarının büyüme için uyarılmış tohum ve çeliklerine orizalin ve N2O gazının farklı doz
ve sürelerde uygulanarak poliploit asma anaçları elde edilmedeki etkinlikleri incelenmiştir. Kimyasal
mutajen uygulamalarının LD50 değerleri belirlenmiş, morfolojik ve sitolojik etkileri makroskobik,
mikroskobik ve sitolojik yöntemlerle incelenmiştir. Tüm mutajen uygulamalarında canlılık oranları
azalttığı gibi, uygulama süresi ve doz artışına zıt bir şekilde LD50 değerleri de azalmıştır. Orizalin ve N2O
uygulamalarıyla uyardığımız poliploit bireylerin yaprak kalınlığı ve klorofil içeriklerinde diploit
ebeveynlerine kıyasla önemli artışlar belirlenmiştir. Benzer şekilde poliploit bireylerin stoma boyutları
önemli ölçüde artarken stoma yoğunlukları azalmış, kloroplast sayılarında önemli artışlar kaydedilmiştir.
Kloroplast ve stoma ölçümleri sonucunda mutant olduğu düşünülen 55 adet örnekte FC analizleri
yapılmıştır. FC analizinde, Fercal neslinden 1 adet tetraploit, 4 adet miksoploit ve 41 B neslinden ise 1
adet miksoploit bitki seçilmiştir. Fercal asma anacı tohumlarına N2O gazının 10 bar 96 saat uygulaması
sunucunda tetraploit elde edilirken miksoploit bitkiler 41 B ve Fercal asma anacının her ikisinde de
orizalinin 100 μM 96 saat uygulaması yapılan fideler arasından seçilmiştir. Miksoploit ve tetraploit
bireylerin nükleer DNA içeriği diploit bitkilerle karşılaştırıldığında sırasıyla 1.34 ve 2.00 kat artmıştır.
Poliploit anaç genotiplerinin kök ucu örneklerinde yapılan kromozom sayımında tetraploit Fercal asma
genotipinin kromozom sayısı diploit orijinaline kıyasla 2 kat arttığı doğrulanmıştır. 41 B ve Fercal asma
anaçlarında farklı doz ve sürelerdeki orizalin ve N2O uygulamaları poliploit bitki uyarımında etkili
bulunmuştur. Uygulamaların poliploidinin doğrulanması, poliploit bireylerin tespitinde ön veriler olarak
kullanılan stoma özellikleri ve kloroplast sayımları kısmen başarılı bulunmuştur. FC analizi miksoploit
genotiperin tespitinde vazgeçilmez olarak görülmektedir. Poliploidi uyarımında elde edilen mutantların
ploidi düzeylerinin tespiti için FC analizi ve kesin doğrulama için kromozom sayımlarının gerekliliği
düşünülmektedir. Bundan sonraki poliploit mutant elde etme çalışmalarında asma anacı genotiplerine,
100 μM-96-saat orizalin ve 10 bar 96-saat N2O dozları öncelikli olarak test edilebilir.
Grape (Vitis spp.) is one of the most important crops globally, both socially and economically. The development of grape varieties and vine rootstocks is of increasing importance to adapt to climate change as well as today's consumer demands. The vine rootstocks obtained by selection and hybridization studies have not been able to meet all demands. Although most grape rootstocks are selected for the European vineyard regions, they cannot fully meet the needs of the producers in the calcareous and shallow soils and vineyard areas with short vegetation in our country. In grape breeding, mutants are more promising than crossbreeding in obtaining new genotypes from natural genetic diversity. Polyploid plants outperform their diploid relatives in several respects. In this study, the effects of oryzalin and N2O gas on seeds and cuttings stimulated for growth of 41 B and Fercal rootstocks were investigated in obtaining polyploid rootstocks by applying different doses and application times. LD50 values of chemical mutagen applications were determined, morphological and cytological effects were examined by macroscopic, microscopic, and cytological methods. Significant increases were determined in leaf thickness and chlorophyll contents of polyploid seedlings, which we stimulated with oryzalin and N2O applications, compared to their diploid parents. Similarly, stomatal sizes of polyploid individuals increased significantly, while their stomatal densities decreased, and significant increases were recorded in chloroplast numbers. FC analyses were performed on 55 samples that were thought to be mutant because of chloroplast and stoma measurements. In FC analyses, 1 tetraploid seedling from Fercal offspring, 4 mixoploid seedlings and 1 mixoploid seedling from 41 B offspring were selected. While tetraploid mutant was obtained by applying N2O gas to Fercal vine rootstock seeds for 96 hours at 10 bar, mixoploid seedlings were selected among the seedlings in which 100 μM-96-hour of oryzalin was applied to both grapevine rootstocks. The nuclear DNA content of mixoploid and tetraploid offspring was increased by 1.34 and 2.00-fold, respectively, when compared to their diploid parents. Chromosome counts of root tip samples of polyploid grapevine rootstock offspring confirmed that the chromosome number of the tetraploid Fercal grapevine rootstock offspring was increased by 2 times compared to the diploid parent. Different doses and durations of oryzalin and N2O applications to 41 B and Fercal rootstocks were effective in stimulating polyploid plants. The stomatal characteristics and chloroplast counts, which were used as preliminary data in the confirmation of the polyploidy of the applications and the detection of polyploid individuals, were found to be partially successful. FC analysis was indispensable in detecting the mixoploid genotype. FC analysis for the detection of ploidy levels of mutants obtained in polyploidy induction and chromosome count were deemed necessary for definitive vi confirmation. In future studies to obtain polyploid mutants in grapevine rootstock genotypes, 100 μM-96- hour oryzalin and 10 bar 96-hour N2O can be tested with priority.
Grape (Vitis spp.) is one of the most important crops globally, both socially and economically. The development of grape varieties and vine rootstocks is of increasing importance to adapt to climate change as well as today's consumer demands. The vine rootstocks obtained by selection and hybridization studies have not been able to meet all demands. Although most grape rootstocks are selected for the European vineyard regions, they cannot fully meet the needs of the producers in the calcareous and shallow soils and vineyard areas with short vegetation in our country. In grape breeding, mutants are more promising than crossbreeding in obtaining new genotypes from natural genetic diversity. Polyploid plants outperform their diploid relatives in several respects. In this study, the effects of oryzalin and N2O gas on seeds and cuttings stimulated for growth of 41 B and Fercal rootstocks were investigated in obtaining polyploid rootstocks by applying different doses and application times. LD50 values of chemical mutagen applications were determined, morphological and cytological effects were examined by macroscopic, microscopic, and cytological methods. Significant increases were determined in leaf thickness and chlorophyll contents of polyploid seedlings, which we stimulated with oryzalin and N2O applications, compared to their diploid parents. Similarly, stomatal sizes of polyploid individuals increased significantly, while their stomatal densities decreased, and significant increases were recorded in chloroplast numbers. FC analyses were performed on 55 samples that were thought to be mutant because of chloroplast and stoma measurements. In FC analyses, 1 tetraploid seedling from Fercal offspring, 4 mixoploid seedlings and 1 mixoploid seedling from 41 B offspring were selected. While tetraploid mutant was obtained by applying N2O gas to Fercal vine rootstock seeds for 96 hours at 10 bar, mixoploid seedlings were selected among the seedlings in which 100 μM-96-hour of oryzalin was applied to both grapevine rootstocks. The nuclear DNA content of mixoploid and tetraploid offspring was increased by 1.34 and 2.00-fold, respectively, when compared to their diploid parents. Chromosome counts of root tip samples of polyploid grapevine rootstock offspring confirmed that the chromosome number of the tetraploid Fercal grapevine rootstock offspring was increased by 2 times compared to the diploid parent. Different doses and durations of oryzalin and N2O applications to 41 B and Fercal rootstocks were effective in stimulating polyploid plants. The stomatal characteristics and chloroplast counts, which were used as preliminary data in the confirmation of the polyploidy of the applications and the detection of polyploid individuals, were found to be partially successful. FC analysis was indispensable in detecting the mixoploid genotype. FC analysis for the detection of ploidy levels of mutants obtained in polyploidy induction and chromosome count were deemed necessary for definitive vi confirmation. In future studies to obtain polyploid mutants in grapevine rootstock genotypes, 100 μM-96- hour oryzalin and 10 bar 96-hour N2O can be tested with priority.
Açıklama
Anahtar Kelimeler
asma anacı, flow sitometri, ıslah, kromozom sayımı, mutajen, poliploidi, breeding, chromosome count, grape rootstock, flow cytometry, mutagen, polyploidy
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Doğan, O., (2022). Azot Protoksit ve Orizalinin ‘41 B’ ile ‘Fercal’ Asma Anaçlarında Mutajenik Etkileri. (Doktora Tezi). Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Konya.
