DDRT-PCR analysis of the expressional modulation showing genes in the maize-maize rust compatible interaction

Abstract

Differential display analyses of expressional modulations occurring in plant-microbe interactions provide valuable information about the physiological changes and underlying genetic factors and mechanisms involved in host resistance and susceptibility to disease. We carried out a differential display analysis study in the compatible interaction of Z. mays and P. sorghi and identified 98 Transcript Derived Fragments (TDFs) as expressional modulation showing tags. 72 TDFs were cloned and sequenced. Sequence database similarity searches revealed that there is at least one close matching GenBank record for each TDF. Compiled results showed that approximately half of the TDFs were derived from genes with known functions, about a half of which are known to display expressional modulations in plant-microbe interactions. CA, BiP2, An2, ARP1 and ASR3-like protein encoding sequence similar TDFs constituted the prominent examples of this group. A large proportion of TDFs were found to be similar to uncharacterized/hypothetical protein encoding GenBank records. RT-qPCR primers were designed for Bip2, CA, cALDH, ARP and Arpp3 genes to verify their observed expressional modulations, which were generally confirmed except for CA. Although a limited number of TDFs were characterized, overall, the results provide an overview to the expressional modulation showing genes in the compatible interaction of the studied pathosystem.

Keywords

• Zea mays
• Puccinia sorghi
• Compatible interaction
• Gene expression
• RT-qPCR

Mısır-mısır pası uyumlu interaksiyonunda ekspresyonu değişim gösteren genlerin DDRT-PCR analizi

Abstract

Bitki-mikrop interaksiyonlarında ekspresyonu değişim gösteren genlerin tanımlanması, konukçu direnci ve duyarlılığında gerçekleşen fizyolojik değişimler ve bunlardan sorumlu genetik faktörler ve mekanizmalar hakkında değerli bilgiler sağlamaktadır. Mısır (Zea mays) ve mısır pası (Puccinia sorghi) uyumlu interaksiyonunda gerçekleştirdiğimiz çalışmada, ekspresyonu değişim gösteren 98 transkript derivativi fragment (TDF) tanımlanmıştır. Klonlanarak sekans karakterizasyonu yapılan 72 TDF ile gerçekleştirilen GenBankası taramaları, münferit etiketler için bir veya daha fazla benzer kayıt bulunduğunu göstermektedir. Genel olarak, TDF’lerin yaklaşık yarısının fonksiyonu bilinen genlerin sekanslarına benzer olduğu ve bunların önemli bir bölümünün bitki-patojen interaksiyonlarında ekspresyonu değişim gösterenler oldukları tespit edilmiştir. Bunlar arasında, karbonik anhidraz, Bip2, An2, ARP ve ASR3 proteinlerini kodlayan genlere benzerlik gösteren TDF’ler bulunmaktadır. TDF’lerin kalan bölümü, diğer stres yanıtlarıyla ilgili olanlar ve karakterize edilmemiş/hipotetik protein kodlayan sekanslara benzerlik göstermektedir. ZmBip2, ZmCA, ZmcALDH, ZmARP ve ZmARPP3 genleri için RT-qPCR primerler tasarlanarak kontrol ve infekte materyalde ekspresyon teyitleri yapıldı. ZmCA hariç, diğerlerinin tespitlerinde gözlenen ekspresyon değişimleri doğrulandı. Sınırlı sayıda TDF çalışılmış olmakla birlikte, belirli fonksiyonlarla ilişkili olanlarla birlikte fonksiyonu bilinmeyenler, çalışılan patosistem uyumlu interaksiyonunda ekspresyonu modülasyon gösteren genler olarak tanımlanmıştır.

Keywords

• Zea mays
• Puccinia sorghi
• uyumlu interaksiyon
• gen ekspresyonu
• RT-qPCR

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