Unresolved issues in pre-meiotic anther development

Kelliher, Timothy; Egger, Rachel L.; Zhang, Han; Walbot, Virginia

doi:10.3389/fpls.2014.00347

Cited by 28 publications

(27 citation statements)

References 42 publications

(74 reference statements)

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“…The result was very similar to fluorescent and colorimetric detection of miR2275 expression in maize anthers at the same stage – that is using SR‐SIM; miR2275 is also mainly localized to the tapetal layer and archesporial cells. We also detected miR2275 in secondary parietal cells, which later give rise to the middle layer and tapetal cells (Kelliher et al ., ) (Figure ). The original STORM method used two dyes and required the dyes to be conjugated to the same antibody.…”

Section: Resultsmentioning

confidence: 98%

sRNA‐FISH: versatile fluorescent in situ detection of small RNAs in plants

2019

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Localization of mRNA and small RNAs (sRNAs) is important for understanding their function. Fluorescent in situ hybridization (FISH) has been used extensively in animal systems to study the localization and expression of sRNAs. However, current methods for fluorescent in situ detection of sRNA in plant tissues are less developed. Here we report a protocol (sRNA-FISH) for efficient fluorescent detection of sRNAs in plants. This protocol is suitable for application in diverse plant species and tissue types. The use of locked nucleic acid probes and antibodies conjugated with different fluorophores allows the detection of two sRNAs in the same sample. Using this method, we have successfully detected the co-localization of miR2275 and a 24-nucleotide phased small interfering RNA in maize anther tapetal and archesporial cells. We describe how to overcome the common problem of the wide range of autofluorescence in embedded plant tissue using linear spectral unmixing on a laser scanning confocal microscope. For highly autofluorescent samples, we show that multi-photon fluorescence excitation microscopy can be used to separate the target sRNA-FISH signal from background autofluorescence. In contrast to colorimetric in situ hybridization, sRNA-FISH signals can be imaged using super-resolution microscopy to examine the subcellular localization of sRNAs. We detected maize miR2275 by super-resolution structured illumination microscopy and direct stochastic optical reconstruction microscopy. In this study, we describe how we overcame the challenges of adapting FISH for imaging in plant tissue and provide a step-by-step sRNA-FISH protocol for studying sRNAs at the cellular and even subcellular level.

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Section: Resultsmentioning

confidence: 98%

sRNA‐FISH: versatile fluorescent in situ detection of small RNAs in plants

2019

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“…Development of male reproduction has been investigated extensively in rice (Oryza sativa) (Raghavan 1988;Zhang and Wilson 2009;Zhang et al 2011), Citrus sinensis (Koltunow et al 1995), Hibiscus syriacus (Kim and Kim 1995), Nelumbo nucifera (Kreunen and Osborn 1999), Arabidopsis (Arabidopsis thaliana) (Sanders et al 1999;Zhang et al 2002;Zhao 2009;Feng and Dickinson 2010;Chang et al 2011), Bromeliaceae (Sajo et al 2005), Onobrychis schahuensis (Chehregani et al 2008), Carthamus tinctorius (Yeung et al 2011), and maize (Zea mays L.) (Kelliher et al 2014). By contrast, few studies on anther development have been performed in the family Araliaceae even though there have been investigations on the morphological aspects of pollen grains.…”

Section: Introductionmentioning

confidence: 99%

“…coordinated (Zhao 2009;Feng and Dickinson 2010;Chang et al 2011;Zhang et al 2011;Kelliher et al 2014;Zhang and Yang 2014).…”

Section: Introductionmentioning

confidence: 99%

Cytological characterization of anther development in Panax ginseng Meyer

et al. 2015

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Ginseng (Panax ginseng), a valued medicinal herb, is a slow-growing plant that flowers after 3 years of growth with the formation of a solitary terminal umbel inflorescence. However, little is known about cytological events during ginseng reproduction, such as the development of the male organ, the stamen. To better understand the mechanism controlling ginseng male reproductive development, here, we investigated the inflorescence and flower structure of ginseng. Moreover, we performed cytological analysis of anther morphogenesis and showed the common and specialized cytological events including the formation of four concentric cell layers surrounding male reproductive cells followed by subsequent cell differentiation and degeneration of tapetal cells, as well as the formation of mature pollen grains via meiosis and mitosis during ginseng anther development. Particularly, our transverse section and microscopic observations showed that the ginseng tapetal layer exhibits obvious nonsynchronous cell division evidenced by the observation of one or two tapetal layers frequently observed in one anther lobe, suggesting the unique control of cell division. To facilitate the future study on ginseng male reproduction, we grouped the anther development into 10 developmental stages according to the characterized cytological events.

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“…Because maize anthers are much larger and develop more slowly , we determined that Ms32 is constitutively expressed starting in early lobe ontogeny, but there are no obvious cytological defects until Stage 9. Arabidopsis DYT1 is expressed across the Arabidopsis anther stages 4-7 (Feng et al, 2012), which corresponds to Stages 4-11 in maize and the rice anther stages 4-8 (Table S6; Sanders et al, 1999;Zhang and Wilson, 2009;Kelliher et al, 2014). Furthermore, Ms32 is expressed in many maize organs (Fig.…”

Section: Impact Of Tp Defects On Small Rna Biogenesismentioning

confidence: 99%

MS23, a master basic helix-loop-helix factor, regulates the specification and development of tapetum in maize

et al. 2016

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Successful male gametogenesis involves orchestration of sequential gene regulation for somatic differentiation in pre-meiotic anthers. We report here the cloning of Male Sterile23 (Ms23), encoding an antherspecific predicted basic helix-loop-helix (bHLH) transcription factor required for tapetal differentiation; transcripts localize initially to the precursor secondary parietal cells then predominantly to daughter tapetal cells. In knockout ms23-ref mutant anthers, five instead of the normal four wall layers are observed. Microarray transcript profiling demonstrates a more severe developmental disruption in ms23-ref than in ms32 anthers, which possess a different bHLH defect. RNA-seq and proteomics data together with yeast two-hybrid assays suggest that MS23 along with MS32, bHLH122 and bHLH51 act sequentially as either homo-or heterodimers to choreograph tapetal development. Among them, MS23 is the earliest-acting factor, upstream of bHLH51 and bHLH122, controlling tapetal specification and maturation. By contrast, MS32 is constitutive and independently regulated and is required later than MS23 in tapetal differentiation.

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Unresolved issues in pre-meiotic anther development

Cited by 28 publications

References 42 publications

sRNA‐FISH: versatile fluorescent in situ detection of small RNAs in plants

sRNA‐FISH: versatile fluorescent in situ detection of small RNAs in plants

Cytological characterization of anther development in Panax ginseng Meyer

MS23, a master basic helix-loop-helix factor, regulates the specification and development of tapetum in maize

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