A Chromosome Preparation Method Using Young Leaves of Citrus.

Kitajima, Akira; Befu, Mayumi; Hidaka, Yoshiko; Hotta, Tomoko; Hasegawa, K.

doi:10.2503/jjshs.70.191

Cited by 6 publications

(10 citation statements)

References 10 publications

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“…Chromosomes in the samples were relatively extended and well-spread. This result agrees with those of previous reports that the enzymatic maceration method is reliable for observing chromosomes of fruit trees (Kitajima et al, 2001;Schuster, 1996;Yamamoto et al, 1999;Zhuang et al, 1990).…”

Section: Discussionsupporting

confidence: 93%

“…Chromosome analysis of some fruit trees has progressed by application of the enzymatic maceration method and/or fluorescent staining (De Melo et al, 2001;Guerra, 1993;Kitajima et al, 2001;Schuster, 1996;Yamamoto et al, 1999;Zhuang et al, 1990). Clearly defined chromosome samples have been obtained by the enzymatic maceration method (Kurata and Omura, 1978).…”

Section: Introductionmentioning

confidence: 99%

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Fluorescent Staining Analysis of Chromosomes in Pear (Pyrus spp.)

Yamamoto

Takada

Hirabayashi

et al. 2010

J. Japan. Soc. Hort. Sci.

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Fluorescent banding patterns of pear chromosomes were determined from samples taken from root tips of openpollinated seedlings of six cultivars from three species [Pyrus pyrifolia (Burm. F) Nakai (Japanese pear), P. communis L. (European pear), and P. bretschneideri Rehder (Chinese pear)]. Root tips were pretreated in 2 mM 8-hydroxyquinoline at 10°C for 4 h; the chromosome samples were prepared by the enzymatic maceration and air-drying method. All cultivars used in this study had 2n = 34 chromosomes. Chromomycin A 3 (CMA) positive (+) bands were observed in telomeric positions of four chromosomes. In some samples, these CMA + bands were observed at satellite positions. 4'-6-diamidino-2-phenylindole (DAPI) negative bands (−) were seen to correspond with CMA + bands. Thirty chromosomes had no CMA +/DAPI − bands. No propidium iodide (PI) bands were observed in any chromosomes. The number and positions of CMA + bands were stable, and there was no difference among the three species.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Fluorescent Staining Analysis of Chromosomes in Pear (Pyrus spp.)

Yamamoto

Takada

Hirabayashi

et al. 2010

J. Japan. Soc. Hort. Sci.

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“…The chromosome preparation method was followed that of Kitajima et al (2001). Young spring leaves, about 3-5 mm long, from trifoliate orange were immersed in 2 mM 8-hydroxyquinoline for 3 h at 20°C and fixed in methanol-acetic acid (3 : 1, v/v).…”

Section: Plant Materials and Chromosome Preparationmentioning

confidence: 99%

Chromosome Identification and Characterization in Trifoliate Orange (Poncirus trifoliata (L.) Raf.) by CMA and PI/DAPI Staining and GISH

バンナラット¹,

宣²,

安津³

et al. 2007

J. Japan. Soc. Hort. Sci.

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The chromosomes of trifoliate orange (Poncirus trifoliata (L.) Raf.) were stained with chromomycin A 3 (CMA) and double stained with alkylating fluorochromes 4'-6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI). PI-positive (PI (+)) regions in PI/DAPI staining were identical to CMA-positive (CMA (+)) regions. The signals of PI (+) were more stable than those of CMA (+). Based on the relative-sized PI (+) signals and the morphology of no-signal chromosomes, eighteen chromosomes of trifoliate orange were classified into 9 groups. Genomic in situ hybridization (GISH) was performed on trifoliate orange chromosomes using the double probe of DNA from trifoliate orange labeled with digoxigenin-rhodamine (red), and from 'Nankan No. 20' satsuma mandarin (Citrus unshiu Marcow.) or 'Tosa Buntan' pummelo (C. maxima (Burm.) Merr.) labeled with biotin-FITC (green). Sixteen GISH signals with a higher intensity were detected in the identical positions of PI (+) regions. The coloration of GISH showed 13 yellow signals and 1 green signal by the biotin probes of satsuma mandarin, and 5 yellow signals and 11 red signals by those of 'Tosa Buntan' pummelo. This indicates that trifoliate orange is more closely related to satsuma mandarin than 'Tosa Buntan' pummelo, because red signals indicate that they share no homologous sequences between genomes. GISH signals on the secondary constriction regions of all B-type chromosomes and on the telomeric region of the D 4 -type chromosome were yellow or green in both satsuma mandarin and 'Tosa Buntan' pummelo biotin probes. This result suggests that these regions in trifoliate orange chromosomes can be homologous to satsuma mandarin and 'Tosa Buntan' pummelo genomes, and are thus conserved regions.

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“…In the case of the Arenaria ciliata complex, as for other species, the alternative is to use shoot meristems (Kitajima et al . ). Tissue disruption and exposure of dividing cells is usually achieved after pretreatment with either enzymatic digestion (Schwarzacher & Leitch ) or hydrochloric acid (Ma et al .…”

Section: Introductionmentioning

confidence: 97%

“…; Kitajima et al . ), followed by Feulgen cell staining and visualization using confocal microscopy (Hepler & Gunning ). Flow cytometry analysis has utilized propidium iodide staining of unknown and calibrated control samples, with samples drawn from living and archival frozen tissue collections (after Galbraith et al .…”

Section: Introductionmentioning

confidence: 99%

Characterization of diverse ploidy in the arctic‐alpine Arenaria ciliata species complex (Caryophyllaceae) using shoot meristem staining and flow cytometry analysis of archived frozen tissue

Abukrees

Kozlowski

Meade

2018

Plant Species Biology

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Ploidy levels were analyzed in 21 European populations of the Arenaria ciliata complex using baseline chromosome counts derived from Feulgen staining of HCl‐treated shoot meristems and calibrated flow‐cytometry analysis of fresh and archival frozen tissue. Calibration with two to three control samples of different ploidy facilitated rapid identification of ploidy states in unknown samples. Observed ploidy levels varied from 2N = 40–200, with the majority of populations showing 2N = 40–80. High‐altitude populations collectively showed the full range of ploidy states, but at low elevations only lower ploidy levels were observed. Populations with the highest observed ploidy contained the greatest observed phylogenetic diversity in the western and eastern Alps. Multiple polyploidization events are inferred in the continental European metapopulation, with lower, more stable ploidy characteristic of the west and north. The method deployed provides an effective approach to ploidy analysis for archival desiccated/frozen tissue samples from biogeographic collections.

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A Chromosome Preparation Method Using Young Leaves of Citrus.

Cited by 6 publications

References 10 publications

Fluorescent Staining Analysis of Chromosomes in Pear (Pyrus spp.)

Fluorescent Staining Analysis of Chromosomes in Pear (Pyrus spp.)

Chromosome Identification and Characterization in Trifoliate Orange (Poncirus trifoliata (L.) Raf.) by CMA and PI/DAPI Staining and GISH

Characterization of diverse ploidy in the arctic‐alpine Arenaria ciliata species complex (Caryophyllaceae) using shoot meristem staining and flow cytometry analysis of archived frozen tissue

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