RETRACTED ARTICLE: Cloning and characterization of TaPUB4 , a U-box gene from common wheat ( Triticum aestivum L.) invol
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Cloning and characterization of TaPUB4, a U-box gene from common wheat (Triticum aestivum L.) involved in regulation of pollen development by influencing sucrose-starch pathway in anther Weidong Zhang & Liang Wang & Yongzhen Wang & Yue Wang & Qingrong Gao Received: 26 September 2019 / Accepted: 18 May 2020 # Springer Nature B.V. 2020
Abstract Development of functional pollen grains in plants is regulated through complex genetic networks and environmental clues. The ubiquitin-26S proteasome system is important in the post-translational modification of intracellular proteins, and the system includes the ubiquitin activating (E1), ubiquitin conjugating (E2), and ubiquitin ligase (E3) enzymes. Here, a novel plant U-box-containing cDNA was isolated in anther from common wheat (Triticum aestivum) and designed as TaPUB4. The cDNA encoding comprised 2598 bp, with an open reading frame of 865 amino acids. The deduced amino acid sequence of TaPUB4 showed 41.2–92.3% identity with other plant Ubox homologous genes. Self-ubiquitination assay indicated that the bacterially expressed TaPUB4 protein had E3 ligase activity, and subcellular localization results showed that TaPUB4 was located in the nucleus and cytoplasm in onion epidermal cells. Overexpression of TaPUB4 gene in Arabidopsis atpub4 mutant restored partially the fertility of the mutant, increased the number of pollens in anthers, enhanced pollen viability, and displayed normal
degradation of tapetum as anther development. The expression levels of genes related to starch deposition also changed greatly in the anthers of transgenic lines. These results support the notion that as a homolog of AtPUB4, TaPUB4 might control male fertility by regulating sucrosestarch metabolism in pollen grains, and this process may be accomplished through ubiquitin-proteasome system. The development and utilization of TaPUB4 as a functional molecular marker may be helpful for improving its fertility in wheat breeding programs in the future. Keywords TaPUB4 . Triticum aestivum . Sterility . E3 ligase . Ubiquitin-proteasome system Abbreviations ARM Armadillo repeats CaMV 35S cauliflower mosaic virus promoter GFP Green fluorescent protein GST Glutathione S-transferase PUB Plant U-box
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11032-020-01138-4) contains supplementary material, which is available to authorized users. W. Zhang : L. Wang : Y. Wang : Y. Wang : Q. Gao State Key Laboratory of Crop Sciences, Shandong Agricultural University, Taian City, Shandong Province, China
W. Zhang : Q. Gao (*) Genetic and Breeding Department, College of Agronomy, Shandong Agricultural University, Taian City, Shandong Province, China e-mail: [email protected]
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qRT-PCR RACE RING E3 UPS WT
Quantitative real-time PCR Rapid amplification of cDNA ends Really interesting new gene Ubiquitin ligase Ubiquitin/26S proteasome systems Wild type
Introduction In higher plants, shoot apical meristem turns into an inflorescence meristem at the end of ve
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