Abstract
The small genome size (740 Mb), short life cycle (3 months) and high economic importance as a food crop legume make chickpea (Cicer arietinum L.) an important system for genomics research. Although several genetic linkage maps using various markers and genomic tools have become available, sequencing efforts and their use are limited in chickpea genomic research. In this study, we explored the genome organization of chickpea by sequencing approximately 500 kb from 11 BAC clones (three representing ascochyta blight resistance QTL1 (ABR-QTL1) and eight randomly selected BAC clones). Our analysis revealed that these sequenced chickpea genomic regions have a gene density of one per 9.2 kb, an average gene length of 2,500 bp, an average of 4.7 exons per gene, with an average exon and intron size of 401 and 316 bp, respectively, and approximately 8.6% repetitive elements. Other features analyzed included exon and intron length, number of exons per gene, protein length and %GC content. Although there are reports on high synteny among legume genomes, the microsynteny between the 500 kb chickpea and available Medicago truncatula genomic sequences varied depending on the region analyzed. The GBrowse-based annotation of these BACs is available at http://www.genome.ou.edu/plants_totals.html. We believe that our work provides significant information that supports a chickpea genome sequencing effort in the future.
Similar content being viewed by others
References
Bodenteich A, Chissoe S, Wang YF, Roe BA (1993) Shotgun cloning as the strategy of choice to generate templates for high-throughput dideoxynucleotide sequencing. In: Venter JC (ed) Automated DNA sequencing and analysis techniques. Academic Press, London, pp 42–50
Buhariwalla HK, Jayashree B, Eshwar K, Crouch JH (2005) Development of ESTs from chickpea roots and their use in diversity analysis of the Cicer genus. BMC Plant Biol 17(5):16
Burge C, Karlin S (1997) Prediction of complete gene structures in human genomic DNA. J Mol Biol 268:78–94
Cannon SB, Sterck L, Rombauts S, Sato S, Cheung F, Gouzy J, Wang X, Mudge J, Vasdewani J, Schiex T, Spannagl M, Monaghan E, Nicholson C, Humphray SJ, Schoof H, Mayer KF, Rogers J, Quétier F, Oldroyd GE, Debellé F, Cook DR, Retzel EF, Roe BA, Town CD, Tabata S, Van de Peer Y, Young ND (2006) Legume genome evolution viewed through the Medicago truncatula and Lotus japonicus genomes. Proc Natl Acad Sci USA 103:14959–14964
Chissoe SL, Bodenteich A, Wang YF, Wang YP, Burian Dennis CSW, Crabtree J, Freeman A, Iyer K, Jian L, Ma Y, McLaury HJ, Pan HQ, Sharan O, Toth S, Wong Z, Zhang G, Heisterkamp N, Groffen J, Roe BA (1995) Sequence and analysis of the human ABL gene, the BCR gene, and regions involved in the Philadelphia chromosomal translocation. Genomics 27:67–82
Choi HK, Mun JH, Kim DJ, Zhu H, Baek JM, Mudge J, Roe B, Ellis N, Doyle J, Kiss GB, Young ND, Cook DR (2004) Estimating genome conservation between crop and model legume species. Proc Natl Acad Sci 101:15289–15294
Corum T, Pang ECK (2005) Isolation and analysis of candidate ascochyta blight defence genes in chickpea. Part I. Generation and analysis of an expressed sequence tag (EST) library. Physiol Mol Plant Pathol 66:192–200
Detter JC, Jett JM, Lucas SM, Dalin E, Arellano AR, Wang M, Nelson JR, Chapman J, Lou Y, Rokhsar D, Hawkins TL, Richardson PM (2002) Isothermal strand-displacement amplification applications for high-throughput genomics. Genomics 80:691–698
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small amount of fresh leaf tissue. Phytochem Bull 19:11–15
Dubcovsky J, Ramakrishna W, SanMiguel PJ, Busso CS, Yan L, Shiloff BA, Bennetzen JL (2001) Comparative sequence analysis of collinear barley and rice bacterial artificial chromosomes. Plant Physiol 125:1342–1353
Ewing B, Green P (1998) Basecalling of automated sequencer traces using phred. II. Error probabilities. Genome Res 8:186–194
Ewing B, Hillier L, Wendl M, Green P (1998) Basecalling of automated sequencer traces using phred. I. Accuracy assessment. Genome Res 8:175–185
Feng Q, Zhang Y, Hao P, Wang S, Fu G, Huang Y, Li Y, Zhu J, Liu Y, Hu X et al (2002) Sequence and analysis of rice chromosome 4. Nature 420:316–320
Gill KS, Gill BS, Endo TR, Taylor T (1996) Identification and high-density mapping of gene-rich regions in chromosome group 1 of wheat. Genetics 144:1883–1891
Goff SA et al (2002) A draft sequence of the rice genome (Oryza sativa L. ssp. japonica). Science 296:92–100
Gordon D, Abajian C, Green P (1998) Consed: a graphical tool for sequence finishing. Genome Res 8:195–202
Jeandet P, Douillet-Breuil AC, Bessis R, Debord S, Sbaghi M, Adrian M (2002) Phytoalexins from the Vitaceae: biosynthesis, phytoalexins gene expression in transgenic plants, antifungal activity and metabolism. J Agric Food Chem 50:2731–2741
Jurka J (2000) Repbase update: a database and an electronic journal of repetitive elements. Trends Genet 16:418–420
Kosambi DD (1944) The estimation of map distance from recombination values. Ann Eugen 12:172–175
Kato T, Sato S, Nakamura Y, Kaneko T, Asamizu E, Tabata S (2003) Structural analysis of a Lotus japonicus genome: V. Sequence features and mapping of sixty-four TAC clones which cover the 6.4 Mb regions of the genome. DNA Res 10:277–285
Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181
Lichtenzveig J, Scheuring C, Dodge J, Abbo S, Zhang HB (2005) Construction of BAC and BIBAC libraries and their applications for generation of SSR markers for genome analysis of chickpea, Cicer arietinum L. Theor Appl Genet 110:492–510
Manly KF (1998) User’s manual for map manager classic and map manager QT
Nelson JC (1997) QGENE: software for maker-based genomic analysis and breeding. Mol Breed 3:239–245
Neumann P, Koblížková A, Navrátilová A, Macas JI (2006) Significant expansion of Vicia pannonica genome size mediated by amplification of a single type of giant retroelement. Genetics 173:1047–1056
Pan HQ, Wang YP, Chissoe SL, Bodenteich A, Wang Z, Iyer K, Clifton SW, Crabtree JS, Roe BA (1994) The complete nucleotide sequence of the SacBII domain of the P1 pAD10-SacBII cloning vector and three cosmid cloning vectors: pTCF, svPHEP, and LAWRIST16. Genetic Anal Techniques Appl 11:181–186
Rajesh PN, Muehlbauer FJ (2008) Discovery and detection of single nucleotide polymorphism (SNP) in coding and genomic sequences in chickpea (Cicer arietinum L.). Euphytica (in press)
Rajesh PN, Coyne C, Meksem K, Sharma KD, Gupta V, Muehlbauer FJ (2004) Construction of a HindIII bacterial artificial chromosome library and its use in identification of clones associated with disease resistance in chickpea. Theor Appl Genet 108:663–669
Rajesh PN, McPhee K, Muehlbauer FJ (2005) Detection of polymorphism using CAPS and dCAPS markers in two chickpea genotypes. Int Chickpea Pigeonpea Newsl 12:4–6
Rakshit S, Winter P, Tekeoglu M, Juarez Muñoz J, Pfaff T, Benko-Iseppon AM, Muehlbauer FJ, Kahl G (2003) DAF marker tightly linked to a major locus for Ascochyta blight resistance in chickpea (Cicer arietinum L.). Euphytica 132:23–30
Roe BA (2004) Shotgun library construction for DNA sequencing in methods in molecular biology, vol 255: Bacterial artificial chromosomes, vol 1: Library construction, physical mapping and sequencing. Human Press, Totowa, pp 171–187
Roe B, Crabtree J, Khan A (1996) In: Rickwood D (eds) DNA isolation and sequencing: essential techniques series. Wiley, New York
Salamov AA, Solovyev VV (2000) Ab initio gene finding in drosophila genomic DNA. Genome Res 10:516–522
Sandhu D, Gill KS (2002) Gene-containing regions of wheat and the other grass genomes. Plant Physiol 128:803–811
Santra DK, Tekeoglu M, Ratnaparkhe MB, Gupta VS, Ranjekar PK, Muehlbauer FJ (2000) Identification and mapping of QTLs conferring resistance to ascochyta blight in chickpea. Crop Sci 40:1606–1612
Sanyal I, Singh AK, Kaushik M, Amla DV (2005) Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) with Bacillus thuringiensis cry-IAc gene for resistance against pod borer insect Helicoverpa armigera. Plant Sci 168:1135–1146
Sarmah BK, Moore A, Tate W, Molvig L, Morton RL, Rees DP, Chiaiese P, Chrispeels MJ, Tabe LM, Higgins TJV (2004) Transgenic chickpea seeds expressing high levels of a bean α-amylase inhibitor. Mol Breeding 14:73–82
Stein L et al (2002) The generic genome browser: a building block for a model organism system database. Genome Res 12:1599–1610
Tekeoglu M, Rajesh PN, Muehlbauer FJ (2002) Integration of sequence tagged microsatellite sites to the chickpea genetic map. Theor Appl Genet 105:847–854
Ware D, Stein L (2003) Comparison of genes among cereals. Curr Opin Plant Biol 6:121–127
Wheeler DL et al (2005) Plant genome resources at the National Center for Biotechnology Information. Plant Physiol 138:1280–1288
Yu O et al (2002) A draft sequence of the rice genome. Science 296:79–91
Yu O, Shi J, Hession AO, Maxwell CA, McGonigle B, Odell JT (2003) Metabolic engineering to increase isoflavone biosynthesis in soybean seed. Phytochemistry 63:753–763
Zhang J, Madden T (1997) PowerBLAST: a new network BLAST application for interactive and automated sequence analysis and annotation. Genome Res 7:649–656
Acknowledgments
PNR wishes to thank McKnight Foundation, USA, for the financial support. MB wishes to thank National Science Foundation, USA, for supporting genome sequencing research. We thank Hongshing Lai and Steve Kenton of bioinformatics group at OU for implementing the Genome Browser for the Chickpea BACs and for their help and advice during their analysis.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by D. A. Hoisington.
P. N. Rajesh and M. O’Bleness have contributed equally.
Rights and permissions
About this article
Cite this article
Rajesh, P.N., O’Bleness, M., Roe, B.A. et al. Analysis of genome organization, composition and microsynteny using 500 kb BAC sequences in chickpea. Theor Appl Genet 117, 449–458 (2008). https://doi.org/10.1007/s00122-008-0789-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-008-0789-0