Abstract
Persimmon (Diospyros kaki Thunb.) is a polyploidy fruit tree species of economic importance to East Asia. Natural astringency loss is an important trait in persimmon breeding programs. Quantitative real-time PCR was used to determine the number of AST/ast alleles for fruit astringency in persimmon (D. kaki Thunb.). To this end, the cultivar Jiro was transformed with one or two copies of a gene encoding NADP-dependent sorbitol-6-phosphate dehydrogenase (S6PDH), which was used as a standard for measuring the allele number of a sequenced marker tightly linked to the recessive ast locus for nonastringency. Primers for markers linked to the AST or ast allele were then used to measure the AST to ast ratio directly in the progeny of a full-sib cross. From determination of the AST to ast ratio and the results of the S6PDH copy number, the number of AST and ast alleles at the AST/ast locus was estimated. This research supported the hypothesis that D. kaki is a hexaploid with six AST and/or ast alleles. In addition to the determination of the allelic status of the AST locus, the application of real-time PCR for confirmation of the ploidy level and allelic composition of target genes in autopolyploids or allopolyploids was demonstrated.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- qRT-PCR:
-
quantitative real-time PCR
References
Bond U, Neal C, Donnelly D, James TC (2004) Aneuploidy and copy number breakpoints in the genome of large yeasts mapped by microarray hybridization. Curr Genet 45:360–370
Burton TL, Husband BC (2001) Fecundity and offspring ploidy in matings among diploid, triploid and tetraploid Chamerion angustifolium (Onagraceae): consequences for tetraploid establishment. Heredity 87:573–582
Carputo D, Barone A, Frusciante L (2000) 2n gametes in the potato: essential ingredients for breeding and germplasm transfer. Theor Appl Genet 101:805–813
Chevreau E, Lespinasse Y, Gallet M (1985) Inheritance of pollen enzymes and polyploidy origin of apple (Malus x domestica Borkh.). Theor Appl Genet 71:268–277
Choi YA, Tao R, Yonemori K, Sugiura A (2003) Physical mapping of 45S rDNA by fluorescent in situ hybridization in persimmon (Diospyros kaki) and its wild relatives. J Hort Sci Biotechnol 78:265–271
D’Surney SJ, Shugart LR, Theodorakis CW (2001) Genetic markers and genotyping methodologies: an overview. Ecotoxicology 10:201–204
Doyle JI, Doyle JL (1987) A rapid DNA isolation procedure for small quantities or fresh leaf tissue of woody plants. BioTechniques 19:11–15
FAO (2007) ProdSTAT database. http://faostat.fao.org. Accessed 10 Oct 2007
Galbraith (2004) Cytometry and plant sciences: a personal retrospective. Cytometry A 58:37–44
Gao M, Tao R, Miura K, Dandekar AM, Sugiura A (2001) Transformation of Japanese persimmon (Diospyros kaki Thunb.) with apple cDNA encoding NADP-dependent sorbitol-6-phosphate dehydrogenase. Plant Sci 160:837–845
Hagberg A, Ellerstorm S (1959) The competition between diploid, tetraploid and aneuploid rye. Theoretical and practical aspect. Hereditas 45:369–415
Hauck NR, Yamane H, Tao R, Iezzoni AF (2006) Accumulation of nonfunctional S-haplotypes results in the breakdown of gametophytic self-incompatibility in tetraploid Prunus. Genetics 172:1191–1198
Henry IM, Dilkes BP, Young K, Watson B, Wu H, Comai L (2005) Aneuploidy and genetic variation in the Arabidopsis thaliana triploid response. Genetics 170:1979–1988
Henry IM, Dilkes BP, Comai L (2006) Molecular karyotyping and aneuploidy detection in Arabidopsis thaliana using quantitative fluorescent polymerase chain reaction. Plant J 48:307–319
Kanayama Y, Mori H, Imaseki H, Yamaki S (1992) Nucleotide sequence of a cDNA encoding NADP-sorbitol-6-phosphate dehydrogenase from apple. Plant Physiol 100:1607–1608
Kanzaki S, Sato A, Yamada M, Yonemori K, Sugiura A (2001) Identification of molecular markers linked to the trait of natural astringency loss of Japanese persimmon (Diospyros kaki) fruit. J Am Soc Hort Sci 126(1):51–55
Kanzaki S, Sato A, Yamada M, Utsunomiya M, Kitajima A, Ikegami A, Yonemori K (2008) RFLP markers for the selection of pollination-constant and non-astringent (PCNA)-type persimmon and examination of the inheritance mode of the markers. J Jpn Soc Hort Sci 77:28–32
Kanzaki S, Yamada M, Sato A, Mitani N, Utsunomiya N, Yonemori K (2009) Conversion of RFLP markers for the selection of pollination-constant and non-astringent type persimmons (Diospyros kaki Thunb.) into PCR-based markers. J Jpn Soc Hort Sci 78:68–73
Lyrene PM, Vorsa N, Ballington JR (2003) Polyploidy and sexual polyploidization in the genus Vaccinium. Euphytica 133:27–36
Maheswaran G, Welander M, Hutchinson JF, Graham MW, Richards D (1992) Transformation of apple rootstock M26 with Agrobacterium-tumefaciens. J Plant Physiol 139:560–568
Namikawa I, Higashi M (1928) On the number of chromosomes in Diospyros kaki L. f. and Diospyros lotus L. Bot Mag (Tokyo) 42:436–438
Ochshorn Y, Bar-Shira A, Jonish A, Yaron Y (2006) Rapid prenatal diagnosis of aneuploidy for chromosome 21, 18, 13, and X by quantitative fluorescence polymerase chain reaction. Fetal Diagn Ther 21(4):326–331
Randolph (1935) Cytogenetics of tetraploid maize. J Agric Res 50:591–605
Rommel M (1961) Aneuploidy, seed set and sterility in artificially induced autotetraploid Hordeum vulgare L. Can J Genet Cytol 3:272–282
Roux N, Toloza A, Radecki Z, Zapata-Arias FJ, Dolezel J (2003) Rapid detection of aneuploidy in Musa using flow cytometry. Plant Cell Rep 21(5):483–490
Scorza R, Cordts JM, Ramming DW, Emershad RL (1995) Transformation of grape (Vitis vinifera L.) zygotic-derived somatic embryos and regeneration of transgenic plants. Plant Cell Rep 14:589–592
Song P, Cai C, Skokut M, Kosegi B, Petolino J (2002) Quantitative real-time PCR as a screening tool for estimating transgene copy number in WHISKERS™-derived transgenic maize. Plant Cell Rep 20:948–954
TaKaRa (2004) RR041A e.v.0408 ver. 2 protocol. http://www.takara-bio.eu/pdf/SYBR.pdf. Accessed 3 April 2008
Tamura M, Tao R, Yonemori K, Utsunomiya N, Sugiura A (1998) Ploidy level and genome size of several Diospyros species. J Jpn Soc Hort Sci 67:306–312
The French–Italian Public Consortium for Grapevine Genome Characterization (2007) The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature 449:463–468
Tsujie T, Takemura M, Kimura T, Koichiro S, Tsutsui T, Ogita K, Ozaki M, Murata Y (2006) Rapid detection of trisomy 21 by gene dosage analysis using quantitative real-time polymerase chain reaction. J Obstet Gynaecol Res 32:368–372
Wang CJ, Harper L, Cande WZ (2006) High-resolution single-copy gene fluorescence in situ hybridization and its use in the construction of a cytogenetic map of maize chromosome 9. Plant Cell 18:529–544
Yamada A, Tao R (2006) High frequency sexual polyploidization observed in hexaploid Japanese persimmon (Diospyros kaki) ‘Fujiwaragosho’. J Hort Sci Biotechnol 81:402–408
Yonemori K, Yamada M, Sugiura A (2000) Persimmon genetics and breeding. Plant Breed Rev 19:191–225
Yonemori K, Honsho C, Kanzaki S, Ino H, Ikegami A, Kitajima A, Sugiura A, Parfitt DE (2008) Sequence analyses of the ITS regions and the matK gene for determining phylogenetic relationships of Diospyros kaki (persimmon) with other wild Diospyros (Ebenaceae) species. Tree Gen Genomes 4:149–158
Zhuang DH, Kitajima A, Ishida M (1990) Chromosome numbers of Diospyros kaki cultivars. J Jpn Soc Hort Sci 59:289–297
Zimmermann B, HolzgreveW WF, Hahn S (2002) Novel real-time quantitative PCR test for trisomy 21. Clin Chem 48:362–363
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by W. Boerjan
Rights and permissions
About this article
Cite this article
Akagi, T., Kanzaki, S., Gao, M. et al. Quantitative real-time PCR to determine allele number for the astringency locus by analysis of a linked marker in Diospyros kaki Thunb. Tree Genetics & Genomes 5, 483–492 (2009). https://doi.org/10.1007/s11295-009-0202-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11295-009-0202-4