Characterization of Lettuce Chromosomes Based on Condensation Patterns and Physical Mapping of 45S and 5S rDNAs Using Fluorescence &lt;i&gt;in situ&lt;/i&gt; Hybridization

Widarmi, Winny Dewi; Kikuchi, Shinichi; Sassa, Hidenori; Koba, Takato

doi:10.1508/cytologia.85.49

Cited by 4 publications

(3 citation statements)

References 24 publications

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“…Koopman et al (1993) applied chromosome banding techniques to discriminate the chromosomes of Lactuca species. Widarmi et al (2019) and Matoba et al (2007) (Mukai et al 1993;Pedersen and Langridge 1997;Jiang and Gill 2006).…”

Section: Introductionmentioning

confidence: 99%

Cytological Variation of (Aag)7 Repeat on Lettuce Chromosomes by Fluorescence in Situ Hybridization (Fish)

Widarmi¹,

Kikuchi

Sassa

et al. 2021

Int. J. Biosci. Biotech.

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Cytological studies using fluorescence in situ hybridization (FISH) technique provides phylogenetical information in closely related taxa and have been widely applied for karyotyping and studying chromosomal organization and evolution in plant species. In the present study, FISH using a microsatellite sequence of (AAG)7 as the probe was performed in order to discriminate the chromosomes in four Lactuca species, i.e., L. sativa, L. serriola, L. saligna, and L. virosa. The experiment was carried out in April to September 2018 at Laboratory of Genetic and Plant Breeding of Breeding of Graduate School of Horticulture, Chiba University, Japan. Different distribution patterns of (AAG)n signals were shown on the chromosomes in the four Lactuca species studied, In L. sativa and L. serriola, FISH with (AAG)7 sequences revealed dispersed distribution patterns with one pair of bright signals, respectively. While in L. saligna and L. virosa, distinct signals with different intensities were observed in two pairs of chromosomes of L. saligna and five pairs of chromosomes of L. virosa. In conclusion, the AAG repeat signals could be used as cytogenetic landmarks for chromosome identification in Lactuca species.

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Section: Introductionmentioning

confidence: 99%

Cytological Variation of (Aag)7 Repeat on Lettuce Chromosomes by Fluorescence in Situ Hybridization (Fish)

Widarmi¹,

Kikuchi

Sassa

et al. 2021

Int. J. Biosci. Biotech.

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“…To study the regulation of chromatin dynamics in living cells, imaging strategies to detect a specific genomic site or gene locus has been developed Matsunaga 2016, Hirakawa andMatsunaga 2016). Classical imaging methods, such as fluorescent in situ hybridization (FISH), have excellent signal-to-noise ratios but require pre-treatment of cells, such as the cell fixation and the DNA denaturation, which prevents the capture of chromatin dynamics (Matsunaga 2016, Matsunaga and Matsunaga 2017, Kikuchi and Iwamoto 2020, Shafiee et al 2020, Suto et al 2020, Widarmi et al 2020, Amiad-Pavlov et al 2021, Hondo and Azumi 2021, Suto et al 2021, Wang and Sheng 2022. The integration of chromatin labeling with live cell imaging can directly observe the dynamics of the specific chromatin in a nucleus.…”

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confidence: 99%

Various Strategies for Improved Signal-to-Noise Ratio in CRISPR-Based Live Cell Imaging

Matsunaga

2023

CYTOLOGIA

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The various behaviors of cells are closely related to the temporal and spatial dynamics of chromatin. Live cell imaging techniques for chromatin can provide insight into gene expression at the three dimension of a nucleus and offer the novel knowledge in gene regulation for medical diagnosis, crop improvement, and environmental remediation. However, traditional chromatin imaging techniques that rely on the insertion of a target gene are unable to directly analyze the chromatin dynamics in living cell nuclei. CRISPR-based live imaging, with its high accuracy, speed, and low perturbation, has emerged as a reliable tool for high-resolution imaging of the specific chromatin in the nucleus. This review paper introduces the benefits of CRISPR-based live imaging for chromatin dynamics, classifies and analyzes various successful schemes used to improve the signal-to-noise ratio of this method, and discusses future trends in the field.

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“…Chromosome specimen preparation is fundamental for several aspects of cytology, including karyotyping, fluorescence in situ hybridization (FISH), and chromosome microdissection (Bhowmick Biplab and Jha 2019, Kono et al 2019, Dash Chandan et al 2020, Nam et al 2020, Saensouk and Saensouk 2020, Samaropoulou et al 2020, Santra et al 2020, Shafiee et al 2020, Sheng et al 2020, Suto et al 2020, Widarmi et al 2020, Wang et al 2021. These techniques revealed differences in chromosome architecture between species, chromosomal abnormalities associated with diseases, and the roles of chromosomes in sex determination.…”

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confidence: 99%

Effective Chromosomal Preparation Protocol for the Dioecious Plant <i>Silene latifolia</i>

et al. 2021

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Cytology requires chromosome specimens, thus, effective preparation methods are needed. Chromosome specimens are frequently prepared from plant root tips. Furthermore, cell cycle synchronization using chemical reagents is applied to obtain a large number of metaphase chromosome specimens. In this study, we focused on the timing of root tip sampling, which is optimal for the preparation of chromosome specimens of the dioecious plant Silene latifolia. The timing was determined from the time seeds were subjected to a germination treatment. Observation of metaphase chromosomes using microscopy revealed that the number of mitotic cells peaked 54 h after the germination treatment. This trend was also observed when the DNA synthesis inhibitor aphidicolin was administered from 24 to 9 h before sampling time points. We used ice-cold treatment for 8, 16, and 32 h as a chromosome condensation method. The 16 h treatment produced suitable chromosome specimens showing satellite ends of chromosomes, whereas the 32 h treatment produced well-condensed chromosome specimens, which were suitable for counting chromosome numbers. Our findings suggest that the timing of root tip sampling is essential for effectively producing plant chromosome specimens.

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Characterization of Lettuce Chromosomes Based on Condensation Patterns and Physical Mapping of 45S and 5S rDNAs Using Fluorescence <i>in situ</i> Hybridization

Cited by 4 publications

References 24 publications

Cytological Variation of (Aag)7 Repeat on Lettuce Chromosomes by Fluorescence in Situ Hybridization (Fish)

Cytological Variation of (Aag)7 Repeat on Lettuce Chromosomes by Fluorescence in Situ Hybridization (Fish)

Various Strategies for Improved Signal-to-Noise Ratio in CRISPR-Based Live Cell Imaging

Effective Chromosomal Preparation Protocol for the Dioecious Plant <i>Silene latifolia</i>

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