ABSTRACT
Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression levels in luffa plants exposed to various simulated abiotic stresses [i.e., cold, drought, heat, salt, H2O2, and abscisic acid (ABA) treatments] were analyzed by qRT-PCR. The stability of the reference gene expression levels was validated using the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. The results indicated that EF-1α was the most stably expressed and suitable reference gene overall and for the heat, cold, and ABA treatments. Additionally, UBQ expression was stable following the salt treatment, whereas TUB was identified as a suitable reference gene for H2O2 and drought treatments. The reliability of the selected reference genes was verified by analyzing the expression of copper/zinc superoxide dismutase (Cu/Zn-SOD) gene in luffa. When the most unstable reference genes were used for data normalizations, the resulting expression patterns had obvious biases when compared with the expression patterns for the most ideal reference genes used alone or combined. These results will be conducive to more accurate quantification of gene expression levels in luffa.
Subject(s)
Adaptation, Physiological/genetics , Genes, Essential , Luffa/genetics , Peptide Elongation Factor 1/genetics , Plant Proteins/genetics , Real-Time Polymerase Chain Reaction/standards , Abscisic Acid/pharmacology , Cold Temperature , Droughts , Gene Expression Profiling , Gene Expression Regulation, Plant , Hot Temperature , Hydrogen Peroxide/pharmacology , Luffa/drug effects , Luffa/metabolism , Peptide Elongation Factor 1/metabolism , Plant Proteins/metabolism , Reference Standards , Salinity , Stress, Physiological/genetics , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolismABSTRACT
The yield and quality of rice are directly impacted by floral organ development in rice. Understanding of the floral development mechanism will be useful in genetic improvement of yield and quality. In this study, a rice mutant palea degradation 2 (pd2) in an indica cultivar '8PW33' was obtained after 60Co γ-ray treatment. Analysis of the mutant showed that, compared to the wild type, plant height, total grain number per panicle, and sword leaf width were significantly increased, but the seed setting rate were significantly decreased. The florets of the mutant exhibited degraded palea and sickle-shaped tortuous lemma. Detail examination using scanning electron microscopy revealed that when epidermis of the vane and lemma were normal, epidermis of the palea were arranged tightly, which might result from degraded palea. Genetic analysis supported that this mutation phenotype was controlled by a single recessive gene. Polymorphic analysis of simple sequence repeat markers demonstrated that PD2 gene is located on chromosome 9. With a larger mapping population and more indel markers, we further mapped PD2 gene between 2 indel markers with a physical region of about 82 kb. Within this region, there is a cloned gene REP1 known to control rice palea development. By comparing the DNA sequences of REP1 from pd2 and 8PW33, in combination with the results of phenotypic analysis, we concluded that PD2 is an allele of REP1.
