Abstract
As a genome model of fruit trees, peach (Prunus persica [L.] Batch) has advantages for studying structural and functional genomics. Okubo, a traditional peach variety used as a parent in Asian peach breeding, displays economically valuable agronomic traits. To develop an efficient platform for peach gene cloning and genomic research, a large-insert genomic DNA library of Okubo was constructed in a transformation-competent artificial chromosome (TAC) vector, pYLTAC7, which can accept and stably maintain large genomic DNA fragments in bothEscherichia coli andAgrobacterium tumefaciens. The TAC library contains 41,472 recombinant clones with an average insert size of approximately 42 kb, and it is equivalent to 6 haploid peach genomes. The TAC library was stored in 2 ways: one copy as frozen cultures in 108 pieces of 384-well plates and another copy as bulked pools in 36 pieces of 96-well plates, each well containing 12 individual clones. The lack of hybridization signal to chloroplast and mitochondrial genes indicated that the TAC library had no significant cytoplast organelle DNA contamination. TAC clones were stable inE. coli cells until generation 100 and stable in bothE. coli andA. tumefaciens. Twenty-one clones containing the polygalacturonase-inhibiting protein (PGIP) gene were detected by using pooled PCR in the TAC library. Positive clones can be used for peach PGIP gene cloning and functional analysis. The library is well suited for gene cloning and genetic engineering in peach.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- BAC:
-
bacterial artificial chromosome
- CIAP:
-
calf intestinal alkaline phosphatase
- EDTA:
-
ethylenediaminetetraacetic acid
- HB:
-
homogenization buffer
- HMW:
-
high molecular weight
- LMP:
-
low melting point
- ORF:
-
open reading frame
- PFGE:
-
pulsed field gel electrophoresis
- PG:
-
polygalacturonase
- PGIP:
-
polygalacturonase-inhibiting protein
- PMSF:
-
phenylmethyl sulfonyl fluoride
- TBE:
-
buffer of Tris-base, boric acid, and EDTA
- TAC:
-
transformation-competent artificial chromosome
References
Abbott AG, Ballard RE, Callahan A, Sosinski BR, Arus P, Wing R, Reighard GL, Baird WV, Parfitt DE, and Dandekar A (2003) Structural and functional genomics in peach: a plant genome model. Plant and Animal Genomes XI Conference, 11–15 January 2003, San Diego, CA, USA.
Arondel V, Lemieux B, Hwang I, Gibson S, Goodman HM, and Somerville CR (1992) Map-based cloning of a gene controlling omega-3 fatty acid desaturation inArabidopsis. Science 258: 1353–5.
Arumuganathan K and Earle E (1991) Nuclear DNA content of some important plant species. Plant Mol Biol Rep 9: 208–18.
Baird WV, Estager AS, and Wells JK (1994) Estimating nuclear DNA content in peach and related diploid species using large flow cytometry and DNA hybridization. J Am Soc Hortic Sci 119: 1312–6.
Belthoff LE, Ballard R, Abbott A, Baird WV, Morgens P, Callahan A, Scorza R, and Monet R (1993) Development of a saturated linkage map ofPrunus persica using molecular based marker systems. Acta Hortic 336: 51–6.
Chapparo JX, Werner DJ, O'Malley D, and Sederoff RR (1994) Targeted mapping and linkage analysis of morphological, isozyme, and RAPD markers in peach. Theor Appl Genet 87: 805–15.
Cheung WY and Gale MD (1990) The isolation of high molecular weight DNA from wheat, barley and rye for analysis by pulse-field gel electrophoresis. Plant Mol Biol 14(5): 881–8.
De Lorenzo G and Ferrari S (2002) Polygalacturonase-inhibiting proteins in defense against phytopathogenic fungi. Curr Opin Plant Biol 5(4): 295–9.
Dirlewanger E, Pronier V, Parvery C, Rothan C, Guye A, and Monet R (1998) Genetic linkage map of peach [Prunus persica (L.) Batsch] using morphological and molecular markers. Theor Appl Genet 97: 888–95.
Dixon MA, Hatzixanthis K, Jones DA, Harrison K, and Jones JDG (1998) The tomato Cf-5 disease resistance gene and six homologs show pronounced allelic variation in leucine-rich repeat copy number. Plant Cell 10: 1915–25.
Fang YD, Liu YG, Wu H, Zhang QY, Chen PD, and Liu DJ (2000) Construction of a transformation-competent artificial chromosome (TAC) library of a wheat-haynaldia villosa translocation line. Chinese J of Biotech 16: 433–6.
Giraudat J, Hauge BM, Valon C, Smalle J, Parcy F, and Goodman HM (1992) Isolation of theArabidopsis ABI3 gene by position cloning. Plant Cell 4: 1251–61.
Jelenkovic G and Harrington E (1972) Morphology of the pachytene chromosomes inPrunus persica. Can J Genet Cytol 14: 317–24.
Liu YG, Liu H, Chen LT, Qiu WH, Zhang QY, Wu H, Yang CY, Su J, Wang ZH, and others (2002) Development of new transformation-competent artificial chromosome vectors and rice genomic libraries for efficient gene cloning. Gene 282: 247–55.
Liu YG, Nagaki K, Fujita M, Kawaura K, Uozumi M, and Ogihara Y (2000) Development of an efficient maintenance and screening system for large-insert genomic DNA libraries of hexaploid wheat in a transformation-competent artificial chromosome (TAC) vector. Plant J 23: 687–95.
Liu YG, Shirano Y, Fukaki H, Yanai Y, Tasaka M, Tabata S, and Shibata D (1999) Complementation of plant mutants with large genomic DNA fragments by a transformation-competent artificial chromosome vector accelerates positional cloning. Proc Natl Acad Sci 96: 6535–40.
Liu YG and Whittier RF (1994) Rapid preparation of megabase plant DNA from nuclei in agarose plugs and microbeads. Nucl Acids Res 22: 2168–9.
Lu ZX, Sonsinsiki B, Reighard GL, Barrd WV, and Abbott AG (1998) Construction of a genetic linkage map and identification of AFLP markers for resistance to root-knot nematodes in peach rootstocks. Genome 41: 199–207.
Lund AH, Duch M, and Pedersen, FS (1996) Increased cloning efficiency by temperature-cycle ligation. Nucl Acids Res 24: 800–1.
Rajapakse S, Belthoff LE, He G, Estager AE, Scorza R, Verde I, Ballard RE, Baird WV, Callahan A, Monet R, and Abbott AG (1995) Genetic linkage mapping in peach using morphological, RFLP and RAPD markers. Theor Appl Genet 91: 964–71.
Sambrook J, Frisch E, and Maniatus T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Scorza R and Sherman WB (1996) Peaches. In: Janik J and Moore JN (eds), Fruiting breeding, vol. 1: tree and tropical fruits, pp 325–440, John Wiley and Sons, Inc., New York.
Shizuya H, Birren B, Kim UJ, Mancino V, Slepak T, Tachiiri Y, and Simon M (1992) Cloning and stable maintenance of 300-kilobase-pair fragments inEscherichia coli using an F-factor-based vector. Proc Natl Acad Sci 89: 8794–7.
Song WY, Wang GL, Chen LL, Kim H, Pi LY, Holsten T, Gardner J, Wang B, Zhai WX, Zhu LH, and others (1995) A receptor kinase-like protein encoded by the rice disease resistance gene Xa21. Science 270: 1804–6.
Sosinski B, Sossey-Alaoui K, Rajapakse S, Glassmoyer K, Ballard RE, Abbott AG, Lu ZX, Baird WV, Reighard G, Tabb A, and Scorza R (1998) Use of AFLP and RFLP markers to create a combined linkage map in peach [Prunus persica (L.) Batsch] for use in marker assisted selection. Acta Hortic 465: 61–68.
Viruel MA, Madur D, Dirlewanger E, Pascal T, and Kervella J (1998) Mapping quantitative trait loci controlling peach leaf curl resistance. Acta Hortic 465: 79–84.
Wang Q, Zhang K, Qu X, Jia J, Shi J, Jin D, and Wang B (2001) Construction and characterization of a bacterial artificial chromosome library of peach. Theor Appl Genet 103: 1174–9.
Wang XP, Zhang ZY, Zhang QY, Liu YG, and Xin ZY (2002) Construction, characterization and screening of a transformation-competent artificial chromosoem (TAC) library of a wheat-thinopyrum intermedium translocation line with resistance to barley yellow dwarf virus. Acta Genetica Sinica 29: 712–8.
Warburton ML, Becerra-Velasquez VL, Goffreda JC, and Bliss FA (1996) Utility of RAPD markers in identifying genetic linkages to genes of economic interest in peach. Theor Appl Genet 93: 920–5.
Woo SS, Rastogi VK, Zhang HB, Paterson AH, Schertz KF, and Wing R (1995) Isolation of megabase-size DNA fromsorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction. Plant Mol Biol Rep 13: 82–94.
Zhang HB, Choi S, Woo SS, Jiang J, Li Z, and Wing RA (1996) Construction and characterization of two rice bacterial artificial chromosome libraries from parents of a permanent recombinant inbred mapping population. Mol Breeding 2: 11–24.
Zhang H, Zhao X, Ding X, Paterson AH, and Wing RA (1995) Preparation of megabase-size DNA from plant nuclei. Plant J 7: 175–84.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liang, F.S., Zhang, K.C., Yu, Z.W. et al. Construction, characterization, and screening of a transformation-competent artificial chromosome library of peach. Plant Mol Biol Rep 22, 37–48 (2004). https://doi.org/10.1007/BF02773347
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF02773347