Transformation of rice with large maize genomic DNA fragments containing high content repetitive sequences
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Large and complex maize BIBAC inserts, even with a length of about 164 kb and repeat sequences of 88.1 %, were transferred into rice.
The BIBAC vector has been established to clone large DNA fragments and directly transfer them into plants. Previously, we have constructed a maize B73 BIBAC library and demonstrated that the BIBAC clones were stable in Agrobacterium. In this study, we demonstrated that the maize BIBAC clones could be used for rice genetic transformation through Agrobacterium-mediated method, although the average transformation efficiency for the BIBAC clones (0.86 %) is much lower than that for generally used binary vectors containing small DNA fragments (15.24 %). The 164-kb B73 genomic DNA insert of the BIBAC clone B2-6 containing five maize gene models and 88.1 % of repetitive sequences was transferred into rice. In 18.75 % (3/16) of the T1, 13.79 % (4/29) of the T2, and 5.26 % (1/19) of the T3 generation transgenic rice plants positive for the GUS and HYG marker genes, all the five maize genes can be detected. To our knowledge, this is the largest and highest content of repeat sequence-containing DNA fragment that was successfully transferred into plants. Gene expression analysis (RT-PCR) showed that the expression of three out of five genes could be detected in the leaves of the transgenic rice plants. Our study showed a potential to massively use maize genome resource for rice breeding by mass transformation of rice with large maize genomic DNA fragment BIBAC clones.
KeywordsBIBAC Large DNA fragment Genetic transformation Agrobacterium Rice Maize
Bacterial artificial chromosome
Basic local alignments search tool
Transformation-competent artificial chromosome
Pulsed-field gel electrophoresis
We thank Dr. Carol M. Hamilton and the Cornell Center for Technology Enterprise and Commercialization for providing the help for vector pCH32 and pCH30. This work was supported by the National Programs for High Technology Research and Development (863 Project: 2012AA10A305) and the National Natural Science Foundation of China (Grant No. 30971748).
Conflict of interest
The authors declare that they have no conflict of interest.
- Andorf CM, Lawrence CJ, Harper LC, Schaeffer ML, Campbell DA, Sen TZ (2010) The Locus Lookup tool at MaizeGDB: identification of genomic regions in maize by integrating sequence information with physical and genetic maps. Bioinformatics 26:434–436. doi: 10.1093/bioinformatics/btp556 CrossRefPubMedGoogle Scholar
- Chang YL, Chuang HW, Meksem K, Wu FC, Chang CY, Zhang M, Zhang HB (2011) Characterization of a plant-transformation-ready large-insert BIBAC library of Arabidopsis and bombardment transformation of a large-insert BIBAC of the library into tobacco Genome. National Research Council Canada 54:437–447. doi: 10.1139/g11-011 Google Scholar
- Harper LC, Schaeffer ML, Thistle J, Gardiner JM, Andorf CM, Campbell DA, Cannon EK, Braun BL, Birkett SM, Lawrence CJ, Sen TZ (2011) The MaizeGDB Genome Browser tutorial: one example of database outreach to biologists via video database. J Biol Databases Curation 2011:bar016. doi: 10.1093/database/bar016 Google Scholar
- Li Y, Uhm T, Ren C, Wu C, Santos TS, Lee MK, Yan B, Santos F, Zhang A, Scheuring C, Sanchez A, Millena AC, Nguyen HT, Kou H, Liu D, Zhang HB (2007) A plant-transformation-competent BIBAC/BAC-based map of rice for functional analysis and genetic engineering of its genomic sequence. Genome National Research Council Canada 50:278–288. doi: 10.1139/g07-006 Google Scholar
- Liu YG, Shirano Y, Fukaki H, Yanai Y, Tasaka M, Tabata S, 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 USA 96:6535–6540CrossRefPubMedCentralPubMedGoogle Scholar
- Ortiz-Vazquez E, Kaemmer D, Zhang HB, Muth J, Rodriguez-Mendiola M, Arias-Castro C, James A (2005a) Construction and characterization of a plant transformation-competent BIBAC library of the black Sigatoka-resistant banana Musa acuminata cv. Tuu Gia (AA). Theor Appl Genet 110:706–713. doi: 10.1007/s00122-004-1896-1 CrossRefPubMedGoogle Scholar
- Ortiz-Vazquez E, Kaemmer D, Zhang HB, Muth J, Rodriguez-Mendiola M, Arias-Castro C, James A (2005b) Construction and characterization of a plant transformation-competent BIBAC library of the black Sigatoka-resistant banana Musa acuminata cv. Tuu Gia (AA). Theor Appl Genet 110:706–713. doi: 10.1007/s00122-004-1896-1 CrossRefPubMedGoogle Scholar
- Schnable PS, Ware D, Fulton RS, Stein JC, Wei FS, Pasternak S, Liang CZ, Zhang JW, Fulton L, Graves TA, Minx P, Reily AD, Courtney L, Kruchowski SS, Tomlinson C, Strong C, Delehaunty K, Fronick C, Courtney B, Rock SM, Belter E, Du FY, Kim K, Abbott RM, Cotton M, Levy A, Marchetto P, Ochoa K, Jackson SM, Gillam B, Chen WZ, Yan L, Higginbotham J, Cardenas M, Waligorski J, Applebaum E, Phelps L, Falcone J, Kanchi K, Thane T, Scimone A, Thane N, Henke J, Wang T, Ruppert J, Shah N, Rotter K, Hodges J, Ingenthron E, Cordes M, Kohlberg S, Sgro J, Delgado B, Mead K, Chinwalla A, Leonard S, Crouse K, Collura K, Kudrna D, Currie J, He RF, Angelova A, Rajasekar S, Mueller T, Lomeli R, Scara G, Ko A, Delaney K, Wissotski M, Lopez G, Campos D, Braidotti M, Ashley E, Golser W, Kim H, Lee S, Lin JK, Dujmic Z, Kim W, Talag J, Zuccolo A, Fan C, Sebastian A, Kramer M, Spiegel L, Nascimento L, Zutavern T, Miller B, Ambroise C, Muller S, Spooner W, Narechania A, Ren LY, Wei S, Kumari S, Faga B, Levy MJ, McMahan L, Van Buren P, Vaughn MW, Ying K, Yeh CT, Emrich SJ, Jia Y, Kalyanaraman A, Hsia AP, Barbazuk WB, Baucom RS, Brutnell TP, Carpita NC, Chaparro C, Chia JM, Deragon JM, Estill JC, Fu Y, Jeddeloh JA, Han YJ, Lee H, Li PH, Lisch DR, Liu SZ, Liu ZJ, Nagel DH, McCann MC, SanMiguel P, Myers AM, Nettleton D, Nguyen J, Penning BW, Ponnala L, Schneider KL, Schwartz DC, Sharma A, Soderlund C, Springer NM, Sun Q, Wang H, Waterman M, Westerman R, Wolfgruber TK, Yang LX, Yu Y, Zhang LF, Zhou SG, Zhu Q, Bennetzen JL, Dawe RK, Jiang JM, Jiang N, Presting GG, Wessler SR, Aluru S, Martienssen RA, Clifton SW, McCombie WR, Wing RA, Wilson, RK (2009) The B73 maize genome: complexity, diversity, and dynamics. Science 326:1112–1115. doi: 10.1126/science.1178534
- Wang WQ, Wu YR, Li Y, Xie JY, Zhang ZH, Deng ZY, Zhang YY, Yang CP, Lai JB, Zhang HW, Bao HY, Tang SY, Yang CW, Gao P, Xia GX, Guo HS, Xie Q (2010) A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes. Plant Mol Biol 72:91–99. doi: 10.1007/s1110300995533 CrossRefPubMedGoogle Scholar
- Wang C, Shi X, Liu L, Li H, Ammiraju JS, Kudrna DA, Xiong W, Wang H, Dai Z, Zheng Y, Lai J, Jin W, Messing J, Bennetzen JL, Wing RA, Luo M (2013) Genomic resources for gene discovery, functional genome annotation, and evolutionary studies of maize and its close relatives. Genetics 195:723–737. doi: 10.1534/genetics.113.157115 CrossRefPubMedCentralPubMedGoogle Scholar
- Zhai JF, Wang Y, Sun CY, Jiang SC, Wang KY, Zhang Y, Zhang HB, Zhang MP (2013) A plant-transformation-competent BIBAC library of ginseng (Panax ginseng C. A. Meyer) for functional genomics research and characterization of genes involved in ginsenoside biosynthesis. Mol Breeding 31:685–692. doi: 10.1007/s11032-012-9826-4 CrossRefGoogle Scholar