Microarray expression analysis of meiosis and microsporogenesis in hexaploid bread wheat

Crismani, Wayne; Baumann, Ute; Sutton, Tim; Shirley, Neil J.; Webster, Tracie; Spangenberg, Germán; Langridge, Peter; Able, Jason A.

doi:10.1186/1471-2164-7-267

Cited by 72 publications

(68 citation statements)

References 56 publications

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“…The signals were especially strong in premeiotic and leptotene anthers, and were concentrated in areas where the meiocytes develop, but not in the connective tissue between anther lobes. The occurrence of Dmc1 expression before the onset of meiosis has been reported before, for example in wheat [14]. In zygotene, the signals were more confined to meiocytes.…”

Section: Resultssupporting

confidence: 64%

The transcriptome landscape of early maize meiosis

et al. 2014

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BackgroundA major step in the higher plant life cycle is the decision to leave the mitotic cell cycle and begin the progression through the meiotic cell cycle that leads to the formation of gametes. The molecular mechanisms that regulate this transition and early meiosis remain largely unknown. To gain insight into gene expression features during the initiation of meiotic recombination, we profiled early prophase I meiocytes from maize (Zea mays) using capillary collection to isolate meiocytes, followed by RNA-seq.ResultsWe detected ~2,000 genes as preferentially expressed during early meiotic prophase, most of them uncharacterized. Functional analysis uncovered the importance of several cellular processes in early meiosis. Processes significantly enriched in isolated meiocytes included proteolysis, protein targeting, chromatin modification and the regulation of redox homeostasis. The most significantly up-regulated processes in meiocytes were processes involved in carbohydrate metabolism. Consistent with this, many mitochondrial genes were up-regulated in meiocytes, including nuclear- and mitochondrial-encoded genes. The data were validated with real-time PCR and in situ hybridization and also used to generate a candidate maize homologue list of known meiotic genes from Arabidopsis.ConclusionsTaken together, we present a high-resolution analysis of the transcriptome landscape in early meiosis of an important crop plant, providing support for choosing genes for detailed characterization of recombination initiation and regulation of early meiosis. Our data also reveal an important connection between meiotic processes and altered/increased energy production.

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Section: Resultssupporting

confidence: 64%

The transcriptome landscape of early maize meiosis

et al. 2014

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“…The quantification of mRNAs is usually achieved by northern blotting or by ribonuclease protection assay (RPA) (Suzuki et al, 2000). These methods are less precise than RT-qPCR (Bustin, 2000), which has emerged as an important technique to compare the expression profiles of target genes in several species, tissues and treatments, and also to validate high-throughput gene-expression profiles (Crismani et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Identification of reference genes for expression analysis by real-time quantitative PCR in drought-stressed soybean

Stolf-Moreira

Lemos

Abdelnoor

et al. 2011

Pesq. agropec. bras.

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-The objective of this work was to validate, by quantitative PCR in real time (RT-qPCR), genes to be used as reference in studies of gene expression in soybean in drought-stressed trials. Four genes commonly used in soybean were evaluated: Gmβ-actin, GmGAPDH, GmLectin and GmRNAr18S. Total RNA was extracted from six samples: three from roots in a hydroponic system with different drought intensities (0, 25, 50, 75 and 100 minutes of water stress), and three from leaves of plants grown in sand with different soil moistures (15, 5 and 2.5% gravimetric humidity). The raw cycle threshold (Ct) data were analyzed, and the efficiency of each primer was calculated for an overall analysis of the Ct range among the different samples. The GeNorm application was used to evaluate the best reference gene, according to its stability. The GmGAPDH was the least stable gene, with the highest mean values of expression stability (M), and the most stable genes, with the lowest M values, were the Gmβ-actin and GmRNAr18S, when both root and leaves samples were tested. These genes can be used in RT-qPCR as reference gene for expression analysis.Index terms: Glycine max, expression stability, RT-qPCR, water deficit. Identificação de genes de referência para análise de expressão por PCR quantitativo em tempo real, em soja submetida à secaResumo -O objetivo deste trabalho foi validar, pela técnica de PCR quantitativo em tempo real (RT-qPCR) genes para serem utilizados como referência em estudos de expressão gênica em soja, em ensaios de estresse hídrico. Foram avaliados quatro genes comumente utilizados em soja: Gmβ-actin, GmGAPDH, GmLectin e GmRNAr18S. O RNA total foi extraído de seis amostras: três amostras de raízes em sistema de hidroponia com diferentes intensidades de déficit hídrico (0, 25, 50, 75 e 100 minutos de estresse hídrico), e três amostras de folhas de plantas cultivadas em areia com diferentes umidades do solo (15, 5 e 2,5% de umidade gravimétrica). Os dados brutos do intervalo "cycle threshold" (Ct) foram analisados, e a eficiência de cada iniciador foi calculada para uma analise da Ct entre as diferentes amostras. A aplicação do programa GeNorm foi utilizada para a avaliação dos melhores genes de referência, de acordo com a estabilidade. O GmGAPDH foi o gene menos estável, com o maior valor médio de estabilidade de expressão (M), e os genes mais estáveis, com menor valor de M, foram o Gmβ-actin e GmRNAr18S, tanto nas amostras de raízes como nas de folhas. Estes genes podem ser usados em RT-qPCR como gens de referência para análises de expressão gênica.Termos para indexação: Glycine max, estabilidade de expressão, RT-qPCR, déficit hídrico.

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“…). In other plant species, such as petunia, wheat and maize, comparisons of transcript profiles were obtained from anthers sampled at different stages of meiosis (Cnudde et al 2006;Crismani et al 2006;Wijeratne et al 2007). Consequently, detailed information on cell-type-specific gene expression is limited.…”

Section: Introductionmentioning

confidence: 99%

Gene expression profiling of Arabidopsis meiocytes

et al. 2011

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Meiosis is a special type of cell division present in all organisms that reproduce by sexual reproduction. It ensures the transition between the sporophytic and gametophytic state and allows gamete production through meiotic recombination and chromosome number reduction. In this paper, we describe a technique for the isolation of Arabidopsis thaliana male meiocytes. From this cellular material, it was then possible to develop large-scale transcriptome studies using CATMA microarrays and thus to obtain an overview of genes expressed during Arabidopsis meiosis. The expression profiles were studied with either stringent statistical criteria or by performing clustering. Both methods resulted in gene clusters enriched in meiosis-specific genes (from 14- to 55-fold). Analysis of these data provided a unique set of genes that will be pivotal to further analysis aimed at understanding the meiotic process.

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Microarray expression analysis of meiosis and microsporogenesis in hexaploid bread wheat

Cited by 72 publications

References 56 publications

The transcriptome landscape of early maize meiosis

The transcriptome landscape of early maize meiosis

Identification of reference genes for expression analysis by real-time quantitative PCR in drought-stressed soybean

Gene expression profiling of Arabidopsis meiocytes

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