Callus regeneration and polyploidy induction of Allium cepa L var. Bima Brebes using oryzalin

Abstract Polyploidy induction could increase shallot bulb-size to raise consumer preference and local shallot productivity. The research aimed to obtain an effective method of polyploidy induction on callus of onion (Allium cepa) var. Bima Brebes. The experiment was consisted of two experimental steps, which were callus induction of onion and polyploid induction of the callus. A 1×1 cm callus was treated by two drops of oryzalin with concentrations 0, 25, 50, 75, 100, and 120 μM. The ploidy level was identified based on morphological trait, stomatal analysis and DNA content using a flow cytometry. The results showed callus diameter, number of green spots, and number of shoots were decreased with increasing oryzalin concentration. The planlet leaves regenerated from oryzalin treated callus were darker than that of control. The flow cytometry analysis showed that planlets with 75 μM oryzalin was tetraploid, had longer and wider stomata than that of the control.

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The genotypic response towards haploid induction in Turkish onion (Allium cepa L.) germplasm

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha48311964 ◽

2020 ◽

Vol 48 (3) ◽

pp. 1176-1184

Author(s):

Faika YARALI KARAKAN

Keyword(s):

Flow Cytometry ◽

Allium Cepa ◽

Ms Medium ◽

Haploid Induction ◽

Flower Buds ◽

Plant Survival ◽

Survival Percentage ◽

Induction Rate ◽

Allium Cepa L ◽

Medium Flow

Haploid induction efficiency of unpollinated flower buds in twenty-six Turkish onion germplasm were determined on two media, Dunstan and Short medium (BDS) and Murashige and Skoog’s medium (MS). On BDS medium, 5850 flower buds were cultured and the induction rate was 2.10% and a plant survival percentage of 1.62% (2 plants). On MS medium, 5850 flower buds were cultured and 175 plantlets were induced; induction rate was 2.99% and plant survival percentage was 13.71% (24 plants). The highest number of plantlets was obtained from genotype Sanliurfa 2; 27 plantlets on BDS medium while 45 plantlets on MS medium. Flow cytometry confirmed that out of the twenty-six plants, 4 plants were haploid (15.38%), 2 mixoploid (7.69%), 18 (69.23%) diploid and 2 tetraploid (7.69%).

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Nuclear DNA amount in pure species and hybrid willows (Salix): a flow cytometric investigation

Canadian Journal of Botany ◽

10.1139/b97-153 ◽

1998 ◽

Vol 76 (1) ◽

pp. 157-165 ◽

Cited By ~ 6

Author(s):

Jérôme Thibault

Keyword(s):

Flow Cytometry ◽

Ploidy Level ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Natural Hybridization ◽

Ploidy Levels ◽

Key Words Flow Cytometry ◽

Dna Values ◽

Dna Amounts

Flow cytometry (FCM) has been used to estimate the nuclear DNA content of 11 Salix species and 5 hybrids. One hundred and sixty nine individuals were studied including 159 individuals from a sequence of 32 communities along a stretch of river in France and 10 individuals from French and English collections for comparison. Isolated nuclei were stained with propidium iodide. FCM was a significantly more practical and rapid technique than that of establishing the karyotype to survey many samples of Salix for variation in ploidy. The 2C DNA amounts for diploid species ranged from 0.76 to 0.98 pg, and tetraploid values ranged from 1.62 to 1.80 pg. The DNA values were consistent with the known ploidy levels. With the exception of a doubtful Salix xquercifolia, ploidy levels and DNA amounts of hybrids were intermediate compared with those of their parents. Intraspecific variation of nuclear DNA values including instrumental variation was low (i.e., 6-11% at the same ploidy level). FCM appeared to be an accurate tool for determination of Salix triploid hybrids. However, it remains limited concerning hybrids from crosses between species of the same ploidy level. Results suggest that natural hybridization might not be frequent in the communities studied, although they have been subject to disturbance. Previous overestimates of hybridization frequency in willows were probably due to misinterpretation of the effects of the environment on Salix spp. morphology; however, the extent and mechanisms of introgression in the genus remain to be further investigated. Key words: flow cytometry, Salix, hybridization, nuclear DNA content, riparian vegetation, disturbance.

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CALLUS INDUCTION AND PLANT REGENERATION IN ONION (ALLIUM CEPA L.)

Acta Horticulturae ◽

10.17660/actahortic.2005.688.43 ◽

2005 ◽

pp. 301-308 ◽

Cited By ~ 2

Author(s):

P.A. Marinangeli ◽

D.C. Zappacosta ◽

N.R. Curvetto ◽

C.R. Galmarini

Keyword(s):

Plant Regeneration ◽

Allium Cepa ◽

Callus Induction ◽

Allium Cepa L

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Chromosome Number, Ploidy Level, and Nuclear DNA Content in 23 Species of Echeveria (Crassulaceae)

Genes ◽

10.3390/genes12121950 ◽

2021 ◽

Vol 12 (12) ◽

pp. 1950

Author(s):

Guadalupe Palomino ◽

Javier Martínez-Ramón ◽

Verónica Cepeda-Cornejo ◽

Miriam Ladd-Otero ◽

Patricia Romero ◽

...

Keyword(s):

Flow Cytometry ◽

Chromosome Number ◽

Ploidy Level ◽

Dna Content ◽

Chromosome Numbers ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Root Tips ◽

Ploidy Levels ◽

Dna Amounts

Echeveria is a polyploid genus with a wide diversity of species and morphologies. The number of species registered for Echeveria is approximately 170; many of them are native to Mexico. This genus is of special interest in cytogenetic research because it has a variety of chromosome numbers and ploidy levels. Additionally, there are no studies concerning nuclear DNA content and the extent of endopolyploidy. This work aims to investigate the cytogenetic characteristics of 23 species of Echeveria collected in 9 states of Mexico, analyzing 2n chromosome numbers, ploidy level, nuclear DNA content, and endopolyploidy levels. Chromosome numbers were obtained from root tips. DNA content was obtained from the leaf parenchyma, which was processed according to the two-step protocol with Otto solutions and propidium iodide as fluorochrome, and then analyzed by flow cytometry. From the 23 species of Echeveria analyzed, 16 species lacked previous reports of 2n chromosome numbers. The 2n chromosome numbers found and analyzed in this research for Echeveria species ranged from 24 to 270. The range of 2C nuclear DNA amounts ranged from 1.26 pg in E. catorce to 7.70 pg in E. roseiflora, while the 1C values were 616 Mbp and 753 Mbp, respectively, for the same species. However, differences in the level of endopolyploidy nuclei were found, corresponding to 4 endocycles (8C, 16C, 32C and 64C) in E. olivacea, E. catorce, E. juarezensis and E. perezcalixii. In contrast, E. longiflora presented 3 endocycles (8C, 16C and 32C) and E. roseiflora presented 2 endocycles (8C and 16C). It has been suggested that polyploidization and diploidization processes, together with the presence of endopolyploidy, allowed Echeveria species to adapt and colonize new adverse environments.

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MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

JOURNAL OF NEOTROPICAL AGRICULTURE ◽

10.32404/rean.v6i1.3203 ◽

2019 ◽

Vol 6 (1) ◽

pp. 1-7

Author(s):

Diego Pandeló José ◽

José Marcello Salabert De Campos ◽

Lyderson Facio Viccini ◽

Emilly Ruas Alkimim ◽

Marcelo De Oliveira Santos

Keyword(s):

Flow Cytometry ◽

Medicinal Plant ◽

Dna Content ◽

Nuclear Dna ◽

Nuclear Dna Content ◽

Naphthalene Acetic Acid ◽

Flow Cytometry Analysis ◽

Ploidy Levels

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.

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Flow Cytometry Analysis in Voided Urine of Patients with Bladder Cancer

Urologia Journal ◽

10.1177/039156039306000207 ◽

1993 ◽

Vol 60 (2) ◽

pp. 162-166

Author(s):

A. Lotto ◽

G. Carluccio ◽

A. Calisti ◽

A. Disperati ◽

E. Capuzzo ◽

...

Keyword(s):

Flow Cytometry ◽

Bladder Cancer ◽

Quantitative Evaluation ◽

Dna Content ◽

Urine Cytology ◽

Urinary Cytology ◽

Flow Cytometry Analysis ◽

Excellent Correlation ◽

First Diagnosis

Flow cytometry is known to be able to give a quantitative evaluation of the DNA of cellular populations (grade of ploidy), as well as to estimate the percentages of phases (S + G2M) providing useful information about the pathology in question and its aggressivity. This method has been applied in diagnosing patients with bladder cancer, using their voided urine and comparing with urine cytology. Our data, from 59 patients, indicate flow cytometry utility in diagnosing bladder cancer; in fact there is an excellent correlation between the urinary cytology and the DNA content in cytometry which increases in higher grade bladder cancer. The sensitivity of CFM is in the range of 92% to 94%, and is superior to that of conventional voided urine cytology (range 64% to 84%). It is felt that cytofluorometric analysis permits a reliable evaluation of voided urine, not only at first diagnosis, but especially during follow-up.

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Determining Ploidy Level and Nuclear DNA Content in Rubus by Flow Cytometry

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.127.5.767 ◽

2002 ◽

Vol 127 (5) ◽

pp. 767-775 ◽

Cited By ~ 37

Author(s):

Rengong Meng ◽

Chad Finn

Keyword(s):

Flow Cytometry ◽

Pacific Northwest ◽

Ploidy Level ◽

Dna Content ◽

Chromosome Numbers ◽

Nuclear Dna ◽

Diploid Species ◽

Nuclear Dna Content ◽

Dna Flow Cytometry ◽

Ploidy Levels

Nuclear DNA flow cytometry was used to differentiate ploidy level and determine nuclear DNA content in Rubus. Nuclei suspensions were prepared from leaf discs of young leaves following published protocols with modifications. DNA was stained with propidium iodide. Measurement of fluorescence of 40 genotypes, whose published ploidy ranged from diploid to dodecaploid, indicated that fluorescence increased with an increase in chromosome number. Ploidy level accounted for 99% of the variation in fluorescence intensity (r2 = 0.99) and variation among ploidy levels was much higher than within ploidy levels. This protocol was used successfully for genotypes representing eight different Rubus subgenera. Rubus ursinus Cham. and Schldl., a native blackberry species in the Pacific Northwest, which has been reported to have 6x, 8x, 9x, 10x, 11x, and 12x forms, was extensively tested. Genotypes of R. ursinus were predominantly 12x, but 6x, 7x, 8x, 9x, 11x, and 13x forms were found as well. Attempts to confirm the 13x estimates with manual counts were unsuccessful. Ploidy level of 103 genotypes in the USDA-ARS breeding program was determined by flow cytometry. Flow cytometry confirmed that genotypes from crosses among 7x and 4x parents had chromosome numbers that must be the result of nonreduced gametes. This technique was effective in differentiating chromosome numbers differing by 1x, but was not able to differentiate aneuploids. Nuclear DNA contents of 21 diploid Rubus species from five subgenera were determined by flow cytometry. Idaeobatus, Chamaebatus, and Anaplobatus were significantly lower in DNA content than those of Rubus and Cylactis. In the Rubus subgenus, R. hispidus and R. canadensis had the lowest DNA content and R. sanctus had the highest DNA content, 0.59 and 0.75 pg, respectively. Idaeobatus had greater variation in DNA content among diploid species than the Rubus subgenus, with the highest being from R. ellipticus (0.69 pg) and lowest from R. illecebrosus (0.47 pg).

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