Chinese Journal OF Rice Science >
Development and Identification of Insect Resistant Transgenic Rice with Cry30Fa1 Gene
Received date: 2015-12-25
Revised date: 2016-01-15
Online published: 2016-05-10
Modified Cry30Fa1, a Bacillus thuringiensis (Bt) gene encoding a high-efficiency insecticidal protein to the brown planthopper(BPH),was introduced into Shuhui 818(R818) by Agrobacterium-mediated genetic transformation and 46 transgene lines were obtained. We confirmed the valid transformation by RT-qRCR and Western blot at transcriptional level and translational level, and fixed the stable expression Cry30Fa1 in homozygosis. The classical pedigree breeding methods were also used to select the important agronomic traits in transgenic lines. Furthermore, we evaluated the resistance to BPH of different lines in growth chamber and field, and found that the R818-Cry30Fal displayed higher resistance than untransformed R818 in seedling stage. We also observed the reduced population of BPH feeding the transgenic lines, suggesting that the transformed Cry30Fa1 enhanced resistibility to BPH in rice. We cultivated new BT protein transgenic rice restorer line R818-Cry30Fal, which may provide new BPH resistance material to three line hybrid rice breeding and enrich BPH resistance germplasm resources in rice.
Hai-peng WANG, Xiao-xi HUANG, Yue-yang LIANG, Jun ZHU, Cui-xia ZHANG, Xiu-mei WANG, Chang-wei GONG, Ai-ping ZHENG, Qi-ming DENG, Shuang-cheng LI, Ling-xia WANG, Ping LI, Shi-quan WANG . Development and Identification of Insect Resistant Transgenic Rice with Cry30Fa1 Gene[J]. Chinese Journal OF Rice Science, 2016 , 30(3) : 256 -264 . DOI: 10.16819/j.1001-7216.2016.5190
| [1] | Dinh H D, Cuong N D, Tai N D, et al.Analysis of rice pest constraints based on survey data at Tan Phu Thanh village, Cantho Province.Omonrice, 2001, 9: 102-106. |
| [2] | Settle W H, Ariawan H, Astuti E T, et al.Managing tropical rice pests through conservation of generalist natural enemies and alternative prey.Ecology,1996, 77(7): 1975-1988. |
| [3] | 杨瑞芳, 白建江, 朴钟泽, 等. 转cry1Ac1基因抗虫水稻的培育. 分子植物育种, 2014, 12(6): 1103-1111. |
| [3] | Yang R F, Bai J J, Piao Z Z, et al.Development of insect-resistant transgenic rice with cry1Ac1 gene.Mol Plant Breeding, 2014, 12(6): 1103-1111.(in Chinese with English abstract) |
| [4] | 胡巍, 李艳芳, 胡侃, 等. 分子标记辅助选择抗褐飞虱基因改良桂农占的BPH抗性. 分子植物育种, 2015, 13(5): 951-960. |
| [4] | Hu W, Li Y F, Hu K, et al.Improving BPH-resistance of rice cultivar Guinongzhan by marker-assisted selection for BPH-resistant genes.Mol Plant Breeding, 2015, 13(5): 951-960. |
| [5] | Swaminathan M S.The Hindu Survey of Indian Agriculture. The Hindu Group of Publications, 1999: 9-16. |
| [6] | 吴振映, 柳絮, 王文英, 等. 利用农杆菌介导法获得转cry2A抗虫基因水稻的研究. 山东农业科学, 2011(5): 1-37. |
| [6] | Wu Z Y, Liu X, Wang W Y, et al.Development of japonica rice with insect-resistant gene Cry2A* by agrobacterium-mediated transformation.Shandong Agric Sci,2011(5): 1-37.(in Chinese with English abstract) |
| [7] | 祁永斌, 叶胜海, 陆艳婷, 等. 转Cry1A(b)基因抗虫水稻的获得及鉴定. 浙江农业学报, 2007, 19(4): 264-267. |
| [7] | Qi Y B, Ye H S, Lu Y T, et al.Development and identification of insect resistant transgenic rice with Cry1A (b) gene.Acta Agric Zhejiang, 2007, 19(4): 264-267.(in Chinese with English abstract) |
| [8] | 于志晶, 刘丽, 李淑芳, 等. 转Cry1C*基因抗虫水稻的培育. 分子植物育种, 2011, 9(6): 702-708. |
| [8] | Yu Z J, Liu L, Li S F, et al.Development of transgenic insect-resistant japonica rice with Cry1C* gene.Mol Plant Breeding, 2011, 9(6): 702-708.(in Chinese with English abstract) |
| [9] | Rao K V, Rathore K S, Hodges T K, et al.Expression of snowdrop lectin( GNA) in transgenic rice plants confers resistance to rice brown planthopper.Plant J, 1998, 15(4): 469-477. |
| [10] | 左示敏, 张平, 祝树德, 等. 水稻转GNA基因后代对褐飞虱的抗性研究. 作物学报, 2004, 30(4): 371-375. |
| [10] | Zuo S M, Zhang P, Zhu S D, et al.Resistance to brown plant hopper in progenies of GNA transgenic rice.Acta Agron Sin,2004, 30(4): 371-375.(in Chinese with English abstract) |
| [11] | 邹良平, 起登凤, 李平, 等. 利用农杆菌介导法将PTA基因导入水稻的研究. 西南农业学报, 2005, 18(3): 229-233. |
| [11] | Zuo L P, Qi D F, Li P, et al.Studies on transformation of the pinellia tenata agglutinin (pta) gene into rice mediated by agrobacterium.Southwest China J Agric Sci,2005, 18(3): 229-233.(in Chinese with English abstract) |
| [12] | 康俊梅, 熊恒硕, 杨青川, 等. 植物抗虫转基因工程研究进展. 生物技术通报, 2008, 1: 14-19. |
| [12] | Kang J M, Xiong H S, Yang Q C, et al.Progress on genetic engineering of plant for insect resistance.Biotechnol Bull,2008, 1: 14-19.(in Chinese with English abstract) |
| [13] | Zhu J, Zheng A P, Wang S Q, et al.Characterization and expression of cry4Cb1 and cry30Ga1 from Bacillus thuringiensis strain HS18-1.J Invert Pathol,2009, 103(3): 200-202. |
| [14] | Guan P, Dai X J, Zhu J, et al.Characterization of the cry54Ab1 operon of Bacillus thuringiensis subsp. sichuansis strain MC28.Biocont Sci Technol,2014, 24(1): 80-89. |
| [15] | Tan F R, Zhu J, Tang J, et al.Cloning and characterization of two novel crystal protein genes,cry54Ab1 and Cry30Fa1, from Bacillus thuringiensis Strain BtMC28.Curr Microbiol,2009, 58(6): 654-659. |
| [16] | 周宗梁, 林智敏, 耿丽丽, 等. 水稻中cry1Ah1基因密码子优化方案的比较. 生物工程学报, 2012, 28(10):1184-1194. |
| [16] | Zhou Z L, Lin Z M, Geng L L, et al.Comparison of codon optimizations of cry1Ah1 gene in rice.Chin J Biotechnol,2012, 28(10):1184-1194.(in Chinese with English abstract) |
| [17] | 李双成, 王世全, 尹福强, 等. 提高农杆菌介导转化水稻效率的因素. 中国水稻科学, 2005, 19(3): 231-237. |
| [17] | Li S C, Wang S Q, Yin F Q, et al.Some factors affecting agrobacterium-mediated transformation frequency in rice.Chin J Rice Sci,2005, 19(3): 231-237.(in Chinese with English abstract) |
| [18] | Heinrichs E A, Medrano E G, Rapusas H R.Genetic evaluation for insect resistance in rice. Manila: IRRI,1985, 72-177. |
| [19] | 刘光杰, 付志红, 沈君辉, 等. 水稻品种对稻飞虱抗性鉴定方法的比较研究. 中国水稻科学, 2002, 16(1): 52-56. |
| [19] | Liu G J, Fu Z H, Shen Q H, et al.Comparative study on evaluation methods for resistance to rice planthoppers (Homoptera: Delphacidae) in rice.Chin J Rice Sci,2002, 16(1): 52-56.(in Chinese with English abstract) |
| [20] | 程海鹏, 朱睦元, 金伟, 等. 植物转基因沉默研究进展. 生物工程进展, 2001, 21(6): 47-49. |
| [20] | Cheng H P, Zhu M Y, Jin W, et al.Advances of researches on the plant transgene silencing.Prog Biotechnol,2001, 21(6): 47-49.(in Chinese with English abstract) |
| [21] | 唐微. 转基因拷贝数对农艺性状的影响. 安徽农业科学, 2008, 36(32):13991-13992. |
| [21] | Tang W.Effects of trans gene copy number on agronomic characters of transformed plants.J Anhui Agric Sci,2008, 36(32): 13991-13992.(in Chinese with English abstract) |
| [22] | Hiwasa-Tanase K, Nyarubona M, Hirai T, et al.High-level accumulation of recombinant miraculin protein in transgenic tomatoes expressing a synthetic miraculin gene with optimized codon usage terminated by the native miraculin terminator.Plant Cell Rep, 2011, 30(1): 113-124. |
| [23] | 王育花, 赵森, 陈芬, 等. 利用实时荧光定量PCR法检测转基因水稻外源基因拷贝数的研究. 生命科学研究, 2007, 11(4): 301-305. |
| [23] | Wang Y H, Zhao S, Chen F, et al.Estimation of the copy number of exogenous gene in transgenic rice by real-time fluorescence quantitative PCR.Life Sci Res, 2007, 11(4): 301-305.(in Chinese with English abstract) |
| [24] | 陈峰, 傅强, 罗举, 等. 苗期抗性不同的水稻品种成株期对褐飞虱的抗性. 中国水稻科学, 2009, 23(2): 201-206. |
| [24] | Chen F, Fu Q, Luo J, et al.Adult stage resistances to brown planthopper, Nilaparvata lugens of rice varieties with different seedling resistances.Chin J Rice Sci, 2009, 23(2): 201-206.(in Chinese with English abstract) |
| [25] | 胡杰, 杨长举, 张庆路, 等. 基因聚合改良杂交稻组合的稻飞虱田间抗性表现. 应用昆虫学报, 2011, 48(5): 1341-1347. |
| [25] | Hu J, Yang C J, Zhang Q L, et al.Resistance of pyramided rice hybrids to brown planthoppers.Chin J Appl Entomol,2011, 48(5): 1341-1347.(in Chinese with English abstract) |
| [26] | 汪秀峰, 叶芬, 李莉, 等. 转基因水稻恢复性及其F1代Bt蛋白的时空表达分析. 分子植物育种, 2014, 12(6): 1077-1081. |
| [26] | Wang X F, Ye F, Li L, et al.Temporal and spatial expression of Bt toxin in transgenic restorer line and its F1 hybrids.Mol Plant Breeding, 2014, 12(6): 1077-1081.(in Chinese with English abstract) |
| [27] | 于志晶, 蔡勤安, 林秀峰, 等. 转基因抗虫水稻中 Bt 蛋白表达量的研究. 安徽农业科学, 2012, 40(6): 3251-3252. |
| [27] | Yu Z J, Cai Q A, Lin X F, et al.Expression of Bt protein in transgenic insect-resistant rice.J Anhui Agric Sci,2012, 40(6): 3251-3252.(in Chinese with English abstract) |
| [28] | 束春娥, 孙以文, 孙洪武. 转基因抗虫棉承受棉铃虫风险压力研究初报. 江苏农业科学, 2000(4): 38-41. |
| [28] | Su C E, Sun Y W, Sun H W.Primary report on studying transgenic insect resistant cotton under risk pressure from Helicoverpa armigera.Jiangsu Agric Sci,2000(4): 38-41.(in Chinese with English abstract) |
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