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| Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into II-32A/B, a Widely Used Cytogenic Male Sterile System |
| LAI Yun-song, HUANG Hai-qing, XU Meng-yun, WANG Liang-chao, ZHANG Xiao-bo, ZHANG Ji-wen , TU Ju-min |
1、Institute of Crop Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China
2、Institute of Pomology & Olericulture, Sichuan Agricultural University, Chengdu 611130, P.R.China
3、Rice Research Institute of Enshi Academy of Agriculture Sciences, Enshi 445000, P.R.China
4、Chinese National Rice Research Institute, Hangzhou 310006, P.R.China |
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摘要 Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis (Bt) δ-endotoxin and provides effective and economic control of lepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR (TAIL-PCR). Bt was located in the promoter region of LOC.Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile (CMS) system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B (maintainer line) and II-32A (male sterile line). Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice.
Abstract Huahui 1 is an elite transgenic male sterile restorer line of wild rice abortive-type that expresses a Bacillus thuringiensis (Bt) δ-endotoxin and provides effective and economic control of lepidopteran insects. To exploit Huahui 1 to develop a new Bt rice, the insertion site of the Bt gene was determined by thermal asymmetric interlaced PCR (TAIL-PCR). Bt was located in the promoter region of LOC.Os10g10360, approximately 5.35 Mb from the telomere of the short arm of chromosome 10. For the first time, a Bt cytoplasmic male sterile (CMS) system was developed by introgressing Bt from Huahui 1. The recipient CMS system used consisted of Indonesia paddy rice-type II-32B (maintainer line) and II-32A (male sterile line). Marker-assisted selection was used to increase selection efficiency in the backcrossing program. In BC5F1, the Bt plant 85015-8 was selected for further analyses, as it had the highest SSR marker homozygosity. In addition, the linkage drag of the foreign Bt gene in 85015-8 was minimized to 8.01-11.46 Mb. The foreign Bt gene was then delivered from 85015-8 into II-32A. The resultant Bt II-32A and Bt II-32B lines were both resistant to lepidopteran in field trials, and agronomic traits were not disturbed. The maintainability of II-32B, and the male sterility and general combining ability of II-32A, were not affected by the Bt introgression. This study demonstrates a simple and fast approach to develop Bt hybrid rice.
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Received: 20 May 2013
Accepted:
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| Fund: Acknowledgements This work was supported by the National High-Tech R&D Program of China (2006AA10Z159). |
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Corresponding Authors:
TU Ju-min, Tel/Fax: +86-571-88982835, E-mail: jtu@zju.edu.cn
E-mail: jtu@zju.edu.cn
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| About author: LAI Yun-song, Tel/Fax: +86-28-86291136, E-mail: ronson521@hotmail.com |
Cite this article:
LAI Yun-song, HUANG Hai-qing, XU Meng-yun, WANG Liang-chao, ZHANG Xiao-bo, ZHANG Ji-wen , TU Ju-min.
2014.
Development of Insect-Resistant Hybrid Rice by Introgressing the Bt Gene from Bt Rice Huahui 1 into II-32A/B, a Widely Used Cytogenic Male Sterile System. Journal of Integrative Agriculture, 13(10): 2081-2090.
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| Adang M J, Staver M J, Rocheleau T A, Leighton J, BarkerR F, Thompson D V. 1985. Characterized full-length andtruncated plasmid clones of the crystal protein of Bacillusthuringiensis subsp. kurstaki HD-73 and their toxicity toManduca sexta Gene, 36, 289-300.Barton K A, Whiteley H R, Yang N S. 1987. Bacillusthuringiensis section sign-endotoxin expressed intransgenic Nicotiana tabacum provides resistance toLepidopteran insects. Plant Physiology, 85, 1103-1109Butaye K M, Goderis I J, Wouters P F, Pues J M, Delaure SL, Broekaert W F, Depicker A, Cammue B P, de BolleM F. 2004. Stable high-level transgene expression inArabidopsis thaliana using gene silencing mutants andmatrix attachment regions. The Plant Journal, 39, 440-449Chen H, Tang W, Xu C, Li X, Lin Y, Zhang Q. 2005.Transgenic indica rice plants harboring a synthetic cry2A*gene of Bacillus thuringiensis exhibit enhanced resistanceagainst lepidopteran rice pests. Theoretical and AppliedGenetics, 111, 1330-1337Frisch M, Bohn M, Melchinger A E. 1999. Comparison ofselection strategies for marker-assisted backcrossing of agene. Crop Science, 39, 1295-1301Frisch M, Melchinger A E. 2005. Selection theory for markerassistedbackcrossing. Genetics, 170, 909-917Fujimoto H, Itoh K, Yamamoto M, Kyozuka J, Shimamoto K.1993. Insect resistant rice generated by introduction of amodified delta-endotoxin gene of Bacillus thuringiensis.Biotechnology (New York), 11, 1151-1155High S M, Cohen M B, Shu Q Y, Altosaar I. 2004. Achievingsuccessful deployment of Bt rice. Trends in Plant Science,9, 286-292Hospital F, Chevalet C, Mulsant P. 1992. Using markers ingene introgression breeding programs. Genetics, 132,1199-1210Hospital F. 2001. Size of donor chromosome segments aroundintrogressed loci and reduction of linkage drag in markerassistedbackcross programs. Genetics, 158, 1363-1379James C. 2012. Global status of commercialized Biotech/GMcrops. ISAAA Brief No. 44. ISAAA, Ithaca, NY.Khush G S. 2005. What it will take to feed 5.0 billion riceconsumers in 2030. Plant Molecular Biology, 59, 1-6Li J, Brunner A M, Meilan R, Strauss S H. 2008. Matrixattachment region elements have small and variableeffects on transgene expression and stability in field-grownPopulus. Plant Biotechnology Journal, 6, 887-896Li J, Xin Y, Yuan L. 2009. Hybrid rice technologydevelopment. Ensuring China’s Food Security. IFPRIDuscussion Paper 00918. International Food PolicyResearch Institute (IFPRI), Washington D.C., USA.Li Z B. 1980. A preliminary discussion about the classificationof male sterile lines of rice in China. Acta AgronomicaSinica, 6, 17-26 (in Chinese)Liu Y G, Whittier R F. 1995. Thermal asymmetric interlacedPCR: automatable amplification and sequencing of insertend fragments from P1 and YAC clones for chromosomewalking. Genomics, 25, 674-681Lu C. 2010. The first approved transgenic rice in China. GMCrops, 1, 113-115Maqbool S B, Christou P. 1999. Multiple traits of agronomicimportance in transgenic indica rice plants: Analysisof transgene integration patterns, expression levels andstability. Molecular Breeding, 5, 471-480Murray M G, Thompson W F. 1980. Rapid isolation of highmolecular weight plant DNA. Nucleic Acids Research, 8,4321-4325Neeraja C N, Maghirang-Rodriguez R, Pamplona A, HeuerS, Collard B C, Septiningsih E M, Vergara G, Sanchez D,Xu K, Ismail A M, Mackill D J. 2007. A marker-assistedbackcross approach for developing submergence-tolerantrice cultivars. Theoretical and Applied Genetics, 115,767-776Perlak F J, Deaton R W, Armstrong T A, Fuchs R L, Sims SR, Greenplate J T, Fischhoff D A. 1990. Insect resistantcotton plants. Biotechnology (New York), 8, 939-943Servin B, Martin O C, Mezard M, Hospital F. 2004. Towarda theory of marker-assisted gene pyramiding. Genetics, 168, 513-523Sheng C F, Wang H T, Gao L D, Xuan W J. 2003. The currentstatus on large scale occurrence of rice stem borers, theirloss estimation and control and protection strategies inChina. Plant Protection, 29, 37-39Sheng X B, Li Z B. 1986. Advance in the studies on themale sterile cytoplasm of hybrid rice in China. ScientiaAgricultura Sinica, 6, 12-16 (in Chinese)Spiker S, Thompson W F. 1996. Nuclear matrix attachmentregions and transgene expression in plants. PlantPhysiology, 110, 15-21Tang W, Lin Y J. 2007. Field experiment of transgenic cry1Abinsect resistant rice. Hereditas (Beijing), 29, 1008-1012 (in Chinese)Temnykh S, DeClerck G, Lukashova A, Lipovich L, CartinhourS, McCouch S. 2001. Computational and experimentalanalysis of microsatellites in rice (Oryza sativa L.):frequency, length variation, transposon associations, andgenetic marker potential. Genome Research, 11, 1441-1452Tu J, Datta K, Alam M F, Fan Y, Khush G S, Datta S K. 1998.Expression and function of a hybrid Bt toxin gene in thetransgenic rice conferring resistance to insect pests. PlantBiotechnology, 15, 195-203Tu J, Datta K, Oliva N, Zhang G, Xu C, Khush G S, Zhang Q,Datta S K. 2003. Site-independently integrated transgenesin the elite restorer rice line Minghui 63 allow removalof a selectable marker from the gene of interest by selfsegregation.Plant Biotechnology Journal, 1, 155-165Tu J, Zhang G, Datta K, Xu C, He Y, Zhang Q, Khush G S,Datta S K. 2000. Field performance of transgenic elitecommercial hybrid rice expressing Bacillus thuringiensisδ-endotoxin. Nature Biotechnology, 18, 1101-1104Ülker B, Allen G C, Thompson W F, Spiker S, Weissinger AK. 1999. A tobacco matrix attachment region reduces theloss of transgene expression in the progeny of transgenictobacco plants. The Plant Journal, 18, 253-263Wang G Y, Zhang H, Xie Y J. 1995. Transfer of Bt-toxinprotein gene into maize and resistance of transgenic plantsto corn borer. Journal of Agricultural Biotechnology, 3,49-53Wunn J, Kloti A, Burkhardt P K, Biswas G C, Launis K,Iglesias V A, Potrykus I. 1996. Transgenic Indica ricebreeding line IR58 expressing a synthetic cryIA(b) genefrom Bacillus thuringiensis provides effective insect pestcontrol. Biotechnology (New York), 14, 171-176Xu S B, Tao Y F, Yang Z Q, Chu J Y. 2002. A simple andrapid methods used for silver staining and gel preservation.Hereditas (Beijing), 24, 335-336 (in Chinese)Yuan G. 2011. Development of hybrid rice and its prospect.Chinese Agricultural Science Bulletin, 27, 6-11Zeng Q C, Zhou K D, Zhu Z, Luo Q. 2000. Current status inthe use of hybrid rice heterosis in China. Chinese Journalof Rice Science, 14, 243-246 (in Chinese)Zheng K, Huang N, Bennett J, Khush G S. 1995. PCR-basedmarker assisted selection in rice breeding. IRRI DiscussionPaper Series No. 12. International Rice Research Institute(IRRI), Manila, the Philippines.Zhu X D, Zhao J, Yan Q Q, Zhou Q M, Peng L J, Xiao M,Zen X D, Peng G D. 2009. Study on combining ability ofyield and related traits of hybrid rice with different typesof cytoplasm. Journal of Hunan Agricultural University(Natural Sciences), 35, 352-356 (in Chinese) |
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