000			02757nab|a22003377a|4500
999			_c62729 _d62721
001			62729
003			MX-TxCIM
005			20240919021229.0
008			200423s2020||||xxk|||p|op||||00||0|eng|d
022			_a2045-2322
024	8		_ahttps://doi.org/10.1038/s41598-020-59580-5
040			_aMX-TxCIM
041			_aeng
100	1		_aGarrido, J. _916486
245	1	0	_aIdentification and validation of reference genes for RT-qPCR normalization in wheat meiosis
260			_aLondon (United Kingdom) : _bNature Publishing Group, _c2020.
500			_aPeer review
500			_aOpen Access
520			_aMeiosis is a specialized type of cell division occurring in sexually reproducing organisms to generate haploid cells known as gametes. In flowering plants, male gametes are produced in anthers, being encased in pollen grains. Understanding the genetic regulation of meiosis key events such as chromosome recognition and pairing, synapsis and recombination, is needed to manipulate chromosome associations for breeding purposes, particularly in important cereal crops like wheat. Reverse transcription-quantitative PCR (RT-qPCR) is widely used to analyse gene expression and to validate the results obtained by other transcriptomic analyses, like RNA-seq. Selection and validation of appropriate reference genes for RT-qPCR normalization is essential to obtain reproducible and accurate expression data. In this work, twelve candidate reference genes were evaluated using the mainstream algorithms geNorm, Normfinder, BestKeeper and ΔCt, then ranked from most to least suitable for normalization with RefFinder. Different sets of reference genes were recommended to normalize gene expression data in anther meiosis of bread and durum wheat, their corresponding genotypes in the absence of the Ph1 locus and for comparative studies among wheat genotypes. Comparisons between meiotic (anthers) and somatic (leaves and roots) wheat tissues were also carried out. To the best of our knowledge, our study provides the first comprehensive list of reference genes for robust RT-qPCR normalization to study differentially expressed genes during male meiosis in wheat in a breeding framework.
546			_aText in English
650		7	_aWheat _gAGROVOC _2 _91310
650		7	_2AGROVOC _98835 _aGene Expression
650		7	_aPlant breeding _gAGROVOC _2 _91203
650		7	_2AGROVOC _916487 _aReverse transcription
650		7	_2AGROVOC _912563 _aPCR
700	1		_aAguilar, M. _916488
700	1		_aPrieto, P. _916489
773	0		_tNature Scientific Reports _gv. 10, art. 2726 _dLondon (United Kingdom) : Nature Publishing Group, 2020. _x2045-2322 _wa58025
856	4		_yClick here to access online _uhttps://doi.org/10.1038/s41598-020-59580-5
942			_cJA _n0 _2ddc