All Issue

2020 Vol.38, Issue 5 Preview Page

Research Article

Phenotypic Characteristics and Karyotype Analysis of Hibiscus sabdariffa var. sabdariffa by Fluorescence in Situ Hybridization (FISH)
Deepo Deen Mohammad1
,
Islam MD Mazharul1
,
Tapce Cherry Ann1
,
Hong-Yul Kim12
,
Ki-Byung Lim12*
1Department of Horticultural Science, Kyungpook National University
2Institute of Agricultural Science and Technology, Kyungpook National University
* Corresponding Author
31 October 2020. pp. 695-704
PDF XML Citation
Abstract

The aim of this research was to ascertain the chromosome number, karyomorphology, distribution of 5S and 18S rDNA loci, individual chromosome length, total chromosome length, DNA content, genome sizes, and phenotypic characteristics of H. sabdariffa var. sabdariffa. The chromosome number and detailed karyotype were analyzed by fluorescence in situ hybridization (FISH) using 5S and 18S rDNA probes. H. sabdariffa var. sabdariffa was tetraploid, and the chromosome number of somatic cells was 72 (2n = 4x = 72). According to FISH analysis, four 5S rDNA loci and ten 18S rDNA loci were detected. The lengths of the somatic chromosomes ranged from 3.53 to 6.88 μm, with a total chromosome length of 97.95 μm, where the total length of short arms and long arms were 37.16 and 60.79 μm respectively. The somatic cells of this species had twelve pairs of metacentric (m) and six pairs of submetacentric (sm) chromosomes. The karyotype formula was K = (2n) = 4x = 72 = 48m + 24sm. In this experiment, DNA content and genome sizes of H. sabdariffa var. sabdariffa were estimated by flow cytometry. 2C-DNA content and 2C genome size were 4.29 pg and 3338.8 Mbp, respectively. The leaf shape, leaf apex, and leaf margin were ovate, acute, and palmatipartite, respectively. The flower colour was purple and single in type. In addition, the diameter of the flower was 7.2 cm and the length of the flower eye was 1.87 cm. These results will contribute to further breeding and cytogenetic studies of H. sabdariffa var. sabdariffa.

Keywords
breeding
cytogenetics
loci
5S rDNA
18S rDNA

References
1
Akpan GA (2000) Cytogenetic characteristics and the breeding system in six Hibiscus species. Theor Appl Genet 100:315-318. doi:10.1007/s001220050041
10.1007/s001220050041
2
Anamthawat-Jónsson KJMICS (2004) Preparation of chromosomes from plant leaf meristems for karyotype analysis and in situ hybridization. Methods Cell Sci 25:91-95. doi:10.1007/s11022-004-5620-y
10.1007/s11022-004-5620-y15801153
3
Andras S, Hartman T, Marshall J, Marchant R, Power J, Cocking E, Davey M (1999) A drop-spreading technique to produce cytoplasm-free mitotic preparations from plants with small chromosomes. Chrom Res 7:641-647. doi:10.1023/A:1009288119851
10.1023/A:100928811985110628665
4
Babajide J, Bodunde J, Salami A (2004) Quality and sensory evaluation of processed calyces of six varieties of Roselle (Hibiscus sabdariffa L.). Nigerian J Hortic Sci 9:110-115. doi:10.4314/njhs.v9i1.3388
10.4314/njhs.v9i1.3388
5
Babalola S, Babalola A, Aworh O (2001) Compositional attributes of the calyces of roselle (Hibiscus sabdariffa L.). J Food Technol Afric 133-134. doi:10.4314/jfta.v6i4.19306
10.4314/jfta.v6i4.19306
6
Chen JH, Wang CJ, Wang CP, Sheu JY, Lin CL, Lin H-HJFC (2013) Hibiscus sabdariffa leaf polyphenolic extract inhibits LDL oxidation and foam cell formation involving up-regulation of LXRα/ABCA1 pathway. Food Chem 141:397-406. doi:10.1016/j.foodchem.2013.03.026
10.1016/j.foodchem.2013.03.02623768373
7
Clydesdale F, Main J, Francis F (1979) Roselle (Hibiscus sabdariffa L.) anthocyanins as colorants for beverages and gelatin desserts. J food protection 42:204-207. doi:10.4315/0362-028X-42.3.204
10.4315/0362-028X-42.3.20430812289
8
Da-Costa-Rocha I, Bonnlaender B, Sievers H, Pischel I, Heinrich MJFc (2014) Hibiscussabdariffa L.-A phytochemical and pharmacological review. 165:424-443. doi:10.1016/j.foodchem.2014.05.002
10.1016/j.foodchem.2014.05.00225038696
9
Dolezel J, Greilhuber J, Suda (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc 2:2233. doi:10.1038/nprot.2007.310
10.1038/nprot.2007.31017853881
10
Faraji MH, Tarkhani AHJJoE (1999) The effect of sour tea (Hibiscus sabdariffa) on essential hypertension. J Ethnopharmacology 65:231-236. doi:10.1016/S0378-8741(98)00157-3
10.1016/S0378-8741(98)00157-3
11
Fasoyiro S, Ashaye O, Adeola A, Samuel F (2005) Chemical and storability of fruit-flavoured (Hibiscus sabdariffa) drinks. World J Agric Sci 1:165-168
12
Gan Y, Liu F, Chen D, Wu Q, Qin Q, Wang C, Li S, Zhang X, Wang Y, et al. (2013) Chromosomal locations of 5S and 45S rDNA in Gossypium genus and its phylogenetic implications revealed by FISH. PLoS ONE 8:e68207. doi:10.1371/journal.pone.0068207
10.1371/journal.pone.006820723826377PMC3695094
13
Hu J, Su Z, Yue B, Zhang Q (2002) Progress of karyotype analysis method in plants research. Journal of Sichuan Teachers College (Natural Science) 23:239-244
14
Hwang Y-J, Kim HH, Kim J-B, Lim K-B (2011) Karyotype analysis of Lilium tigrinum by FISH. Hortic Environ Biotechnol 52:292-297. doi:10.1007/s13580-011-0225-2
10.1007/s13580-011-0225-2
15
Jo YK, MazharuI MD, Kim C-K, Kim H-Y, Lim K-B (2019) Morphological Characteristics and FISH Analysis of Hibiscus F1 Hybrids and Parental Lines. Korean J Hortic sci Technol 37:630-639
16
Kim K-H, Lee C-S, Kang H-C (2016) Study on the selection of promising cultivars with unique flower characteristics in the recently developed cultivars of rose of sharon (Hibiscus spp.) for landscape uses. J Korean Inst Tradit Landsc Archit 34:124-139. doi:10.14700/KITLA.2016.34.3.124
10.14700/KITLA.2016.34.3.124
17
Knight CA, Molinari NA, Petrov DA (2005) The large genome constraint hypothesis: Evolution, ecology and phonotype. Ann. Bot 95:177-190. doi:10.1093/aob/mci011
10.1093/aob/mci01115596465PMC4246716
18
Konar S, Karmakar J, Ray A, Adhikari S, Bandyopadhyay TK (2018) Regeneration of plantlets through somatic embryogenesis from root derived calli of Hibiscus sabdariffa L.(Roselle) and assessment of genetic stability by flow cytometry and ISSR analysis. PLoS ONE 13:e0202324. doi:10.1371/journal.pone.0202324
10.1371/journal.pone.020232430133505PMC6105006
19
Lattier JD, Chen H, Contreras RN (2019) Variation in Genome Size, Ploidy, Stomata, and rDNA Signals in Althea. J Am Soc Hortic Sci 144:130-140. doi:10.21273/JASHS04618-18
10.21273/JASHS04618-18
20
Levan A, Fredga K, Sandberg AAJH (1964) Nomenclature for centromeric position on chromosomes. Hereditas 52:201-220. doi:10.1111/j.1601-5223.1964.tb01953.x
10.1111/j.1601-5223.1964.tb01953.x
21
Li Y-P, Zhang X, Wu W-T, Miao S-X, Chang J-l (2015a) Chromosome and karyotype analysis of Hibiscus mutabilis f. mutabilis. Front Life Sci 8:300-304. doi:10.1080/21553769.2015.1041166
10.1080/21553769.2015.1041166
22
Li YP, Miao SX, Luo FX (2015b) Chromosome and Karyotype Analysis of Hibiscus syriacus. In Applied Mechanics and Materials, Vol 737. Trans Tech Publ, pp 340-344. doi:10.4028/www.scientific.net/AMM.737.340
10.4028/www.scientific.net/AMM.737.340
23
Liang G, Chen HJPo (2015) Scaling chromosomes for an evolutionary karyotype: a chromosomal tradeoff between size and number across woody species. PLoS ONE 10:e0144669. doi:10.1371/journal.pone.0144669
10.1371/journal.pone.014466926657837PMC4684206
24
Lim K-B, Chung J-D, Van Kronenburg BC, Ramanna MS, De Jong JH, Van Tuyl JMJCR (2000) Introgression of Lilium rubellum Baker chromosomes into L. longiflorum Thunb.: a genome painting study of the F1 hybrid, BC1 and BC2 progenies. Chrom Res 8:119-125. doi:10.1023/A:1009290418889
10.1023/A:100929041888910780700
25
Lim K-B, Wennekes J, Jong JHd, Jacobsen E, Van Tuyl JMJG (2001) Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation. Genome 44:911-918. doi:10.1139/g01-066
10.1139/g01-06611681616
26
Mahadevan N, Kamboj P (2009) Hibiscus sabdariffa Linn.-an overview. Natural Product Radiance 8:77-83
27
Pellicer J, Fay MF, Leitch IJ (2010) The largest eukaryotic genome of all Linn. Societ Bot J 164:10-15. doi:10.1111/j.1095-8339.2010.01072.x
10.1111/j.1095-8339.2010.01072.x
28
Prado E, Faivre-Rampant P, Schneider C, Darmency MJG (1996) Detection of a variable number of ribosomal DNA loci by fluorescent in situ hybridization in Populus species. Genome 39:1020-1026. doi:10.1139/g96-127
10.1139/g96-12718469951
29
Qi Y, Chin KL, Malekian F, Berhane M, Gager J (2005) Biological characteristics, nutritional and medicinal value of roselle, Hibiscus sabdariffa. Circular-urban forestry natural resources and environment 604:1-2
30
Rudkin GT, Stollar B (1977) High resolution detection of DNA-RNA hybrids in situ by indirect immunofluorescence. Nature 265:472. doi:10.1038/265472a0
10.1038/265472a0401954
31
Sánchez-Mendoza J, Domínguez-López A, Navarro-Galindo S, López-Sandoval JAJJofe (2008) Some physical properties of Roselle (Hibiscus sabdariffa L.) seeds as a function of moisture content. J Food Eng 87:391-397. doi:10.1016/j.jfoodeng.2007.12.023
10.1016/j.jfoodeng.2007.12.023
32
Shi L-R, Li H-f, Gao R-Y, Han H-Q (2009) Chromosome Number and Karyotype Analysis of Althaea rosea [J]. Jiangsu Agri Sci 5:173-174
33
Silva GS, Souza MM, de Melo CAF, Urdampilleta JD, Forni-Martins ERJBg (2018) Identification and characterization of karyotype in Passiflora hybrids using FISH and GISH. BMC Genet 19:26. doi:10.1186/s12863-018-0612-0
10.1186/s12863-018-0612-029703134PMC5921547
34
Skovsted AJP (1941) Chromosome numbers in the Malvaceae. II. Compt. Rend. Trav. Lab. Carlsberg, Ser. 23:195-242
35
Song J-J (2001) Studies on the Chromosome Number, Ploidy Relationship and Relative Relationship of Several Plants in Hibisucus [D]. Shantou University
36
Speicher MR, Carter NP (2005) The new cytogenetics: blurring the boundaries with molecular biology. Nat Rev Genet 6:782. doi:10.1038/nrg1692
10.1038/nrg169216145555
37
Stewart RN (1947) The morphology of somatic chromosomes in Lilium. Am J Bot 9-26. doi:10.1002/j.1537-2197.1947.tb12953.x
10.1002/j.1537-2197.1947.tb12953.x20282980
38
Tabassum N, Rafique U, Balkhair KS, Ashraf MA (2014) Chemodynamics of methyl parathion and ethyl parathion: adsorption models for sustainable agriculture. BioMed Res intern. doi:10.1155/2014/831989
10.1155/2014/83198924689059PMC3932840
39
Trask BJ (2002) Human genetics and disease: human cytogenetics: 46 chromosomes, 46 years and counting. Nat Rev Genet 3:769. doi:10.1038/nrg905
10.1038/nrg90512360235
40
Van Laere K, Khrustaleva L, Van Huylenbroeck J, Van Bockstaele E (2010) Application of GISH to characterize woody ornamental hybrids with small genomes and chromosomes. Plant breeding 129:442-447. doi:10.1111/j.1439-0523.2009.01692.x
10.1111/j.1439-0523.2009.01692.x
41
Wong P-K, Yusof S, Ghazali H, Che Man Y (2002) Physico-chemical characteristics of roselle (Hibiscus sabdariffa L.). Nutri Food Sci 32:68-73. doi:10.1108/00346650210416994
10.1108/00346650210416994
42
Yan S, An Y, Sun R, Li S (2008) Research progress of karyotype analysis and chromosome micro-dissection technology. Biotechnol Bulletin 4:70-74
43
Yang D-W, Zhu W-Y, Lin J, ZHAO L, ZHANG Y-D, ZHU Z, CHEN T, WANG C-l (2008) Mapping of QTLs for phenol reaction using chromosome segment substitution lines in rice (Oryza sativa L.). Jiangsu J of Agr Sci 24:751-755
44
Zhu W-Y, Wang C-l (2008) A Review on Chromosomal Segment Substitution Lines in Crops [J]. Jiangsu J Agric Sci 6
Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :원예과학기술지
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 38
  • No :5
  • Pages :695-704
  • Received Date : 2020-04-02
  • Revised Date : 2020-06-17
  • Accepted Date : 2020-07-31
  • DOI :https://doi.org/10.7235/HORT.20200063
Journal Informaiton Horticultural Science and Technology Horticultural Science and Technology
Online Submission
  • scopus
  • NRF
  • KOFST
  • crossref crossmark
  • crossref cited-by
  • crosscheck
  • orcid
  • open access
  • ccl
Journal Informaiton Journal Informaiton - close