Efficiencies of Generating Selectable Marker-Free Transgenic Rice with Different Transformation Methods

doi:10.1016/S1672-6308(08)60088-8

Abstract

Abstract: To study the efficiency of generating selectable marker-free (SMF) transgenic rice, two transformation methods were employed for four rice varieties (Wuxiangjing 9, Longtefu, Xieqingzao and Zhenshan 97). One method is by using a single twin T-DNA binary vector pYH592 in one Agrobacterium strain, which is composed of two separate T-DNA regions (one carrying an antisense Wx gene and the other carrying a HPT gene). The other one, named as two-strain/ two-vector system, is by using two separate binary vectors in two separate Agrobacterium cultures. The results indicated that the average co-transformation frequencies of the antisense Wx gene and the HPT gene were 10.1% and 45.0%, respectively, for the four rice varieties. And the SMF transgenic plants selected from the offsprings of co-transformants were 55.6% and 60.0% in the two-strain/two-vector and twin T-DNA vector binary systems, respectively.

Key words: rice, Agrobacterium tumeficien, selectable marker-free, co-transformation

YU Heng-xiu, LU Mei-fang, CHEN Xiu-hua, GONG Zhi-yun, LIU Qiao-quan, GU Ming-hong. Efficiencies of Generating Selectable Marker-Free Transgenic Rice with Different Transformation Methods[J]. RICE SCIENCE, 2009, 16(4): 254-260 .

References

Li H Q, Li M R. Discussion on factors influencing Agrobacterium-mediated plant transformation. Plant Physiol Comm, 1999, 35: 145–151. (in Chinese with English abstract)
2 Chan M T, Chang H H, Ho S L, Tong W F, Yu S M. Agrobacterium-mediated production of transgenic rice plants expressing a chimeric alpha-amylase promoter/beta-glucuronidase gene. Plant Mol Biol, 1993, 22: 491–506.
3 Hiei Y, Ohta S, Komari T, Kumashiro T. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 1994, 6: 271–282.
4 Rashid H, Yokoi S, Toriyama K, Hinata K. Transgenic plant production mediated by Agrobacterium in indica rice. Plant Cell Rep, 1996, 15: 727–730.
5 Liu Q Q, Zhang J L, Wang Z Y, Hong M M, Gu M H. A highly efficient transformation system mediated by Agrobacterium tumefaciens in rice. Acta Phytophysiol Sin, 1998, 24: 259–271. (in Chinese with English abstract)
6 Yoder J I, Goldsbrough A P. Transformation system for generating marker-free transgenic plants. Biotechnology, 1994, 12: 263–267.
7 Jia S R. Safety evaluation of marker genes in transgenic food plants. Sci Agric Sin, 1997, 30(2): 1–15. (in Chinese with English abstract)
8 Miki B, McHugh S. Selectable marker genes in transgenic plants: Applications, alternatives and biosafety. J Biotechnol, 2004, 107: 193–232.
9 Chen S B, Li X G, Wang F, Zhu Z. Culturing marker-free transgenic plants. China Biotechnol, 2005, 25: 1–7. (in Chinese with English abstract)
10 Cotsaftis O, Sallaud C, Breitler J C, Meynard D, Greco R, Pereira A, Guiderdoni E. Transposon-mediated generation of T-DNA and marker-free rice plants expressing a Bt endotoxin gene. Mol Breeding, 2002, 10: 165–182.
11 Lyznik L A, Mitchell J C, Hirayama L, Hodges T K. Activity of yeast FLP recombinase in maize and rice protoplasts. Nucl Acids Res, 1993, 21(4): 969–975.
12 Komari T, Hiei Y, Saito Y, Murai N, Kumashiro T. Vectors carrying two separate T-DNAs for co-transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers. Plant J, 1996, 10: 165–174.
13 de Neve M, de Buck S, Jacobs A, van Montagu M, Depicker A. T-DNA integration patterns in co-transformed plant cells suggest that T-DNA repeats originate from co-integration of separate T-DNAs. Plant J, 1997, 11: 15–29.
14 de Buck S, Jacobs A, van Montagu M, Depicker A, de Buck S, van Montagu M. The DNA sequences of T-DNA junctions suggest that complex T-DNA loci are formed by a recombination process resembling T-DNA integration. Plant J, 1999, 20: 295–304.
15 Afolabi A S, Worland B, Snape J W, Vain P. A large-scale study of rice plants transformed with different T-DNAs provides new insights into locus composition and T-DNA linkage configurations. Theor Appl Genet, 2004, 109: 815– 826.
16 Breitler J C, Meynard D, Boxtel J V, Royer M, Bonnot F, Cambillau L, Guiderdoni E. A novel two T-DNA binary vector allows efficient generation of marker-free transgenic plants in three elite cultivars of rice (Oryza sativa L.). Transgenic Res, 2004, 13(3): 271–278.
17 Park J, Lee Y K, Kang B K, Chung W I. Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants. Theor Appl Genet, 2004, 109(8): 1562–1567.
18 Parkhi V, Rai M, Tan J, Oliva N, Rehana S, Bandyopadhyay A, Torrizo L, Ghole V, Datta K, Datta S K. Molecular characterization of marker-free transgenic lines of indica rice that accumulate carotenoids in seed endosperm. Mol Genet Gen, 2005, 274: 325–336.
19 Hashizume F, Nakazaki T, Tsuchiya T, Matsuda T. Effectiveness of genotype-based selection in the production of marker free and genetically fixed transgenic lineages: Ectopic expression of a pistil chitinase gene increases leaf-chitinase activity in transgenic rice plants without hygromycin- resistance gene. Plant Biotechnol, 2006, 23(4): 349–356.
20 Darbani B, Eimanifar A, Stewart C N, Camargo W N. Methods to produce marker-free transgenic plants. Biotechnol J, 2007, 2: 83–90.
21 Yu H X, Liu Q Q, Wang L, Zhao Z P, Xu L H, Huang B L, Gong Z Y, Tang S Z, Gu M H. Breeding of selectable marker-free transgenic rice lines containing AP1 gene with enhanced disease resistance. Agric Sci China, 2006, 5(11): 812–819.
22 Sripriya R, Raghupathy V, Veluthambi K. Generation of selectable marker-free sheath blight resistant transgenic rice plants by efficient co-transformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain. Plant Cell Rep, 2008, 27: 1635–1644.
23 Hofgen R, Willmitzer L. Storage of competent cells for Agrobacterium transformation. Nucl Acids Res, 1988, 16: 9877
24 Chen X H, Liu Q Q，Wang Z Y, Zhang J L, Hong M M, Gu M H. Study on some factors influencing Agrobacterium- mediated transformation of indica rice. Jiangsu Agric Res, 2001, 22: 1–6. (in Chinese with English abstract)
25 Yu H X, Liu Q Q, Wang L, Huang J, Gong Z Y, Tang S Z, Wang W C, Li N, Gu M H. Optimization of Agrobacterium- mediated transformation of rice mature embryos and regeneration of transgenic plants with metr gene. Hereditas (Beijing), 2005, 27(5): 756–762. (in Chinese with English abstract)
26 Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucl Acid Res, 1980, 8: 4321–4325.
27 Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. New York, USA: Cold Spring Harbor Laboratory Press, 1989.
28 Lu M F, Liu Q Q, Yu H X, Gu M H. A study on several factors on influencing co-transformation of rice with Agrobactericum binary vectors. Biotechnol Bull, 2005(5): 55–62. (in Chinese with English abstract)

[1]	Prathap V, Suresh KUMAR, Nand Lal MEENA, Chirag MAHESHWARI, Monika DALAL, Aruna TYAGI. Phosphorus Starvation Tolerance in Rice Through a Combined Physiological, Biochemical and Proteome Analysis [J]. Rice Science, 2023, 30(6): 8-.
[2]	Serena REGGI, Elisabetta ONELLI, Alessandra MOSCATELLI, Nadia STROPPA, Matteo Dell’ANNO, Kiril PERFANOV, Luciana ROSSI. Seed-Specific Expression of Apolipoprotein A-I_Milano Dimer in Rice Engineered Lines [J]. Rice Science, 2023, 30(6): 6-.
[3]	Sundus ZAFAR, XU Jianlong. Recent Advances to Enhance Nutritional Quality of Rice [J]. Rice Science, 2023, 30(6): 4-.
[4]	Kankunlanach KHAMPUANG, Nanthana CHAIWONG, Atilla YAZICI, Baris DEMIRER, Ismail CAKMAK, Chanakan PROM-U-THAI. Effect of Sulfur Fertilization on Productivity and Grain Zinc Yield of Rice Grown under Low and Adequate Soil Zinc Applications [J]. Rice Science, 2023, 30(6): 9-.
[5]	FAN Fengfeng, CAI Meng, LUO Xiong, LIU Manman, YUAN Huanran, CHENG Mingxing, Ayaz AHMAD, LI Nengwu, LI Shaoqing. Novel QTLs from Wild Rice Oryza longistaminata Confer Rice Strong Tolerance to High Temperature at Seedling Stage [J]. Rice Science, 2023, 30(6): 14-.
[6]	LIN Shaodan, YAO Yue, LI Jiayi, LI Xiaobin, MA Jie, WENG Haiyong, CHENG Zuxin, YE Dapeng. Application of UAV-Based Imaging and Deep Learning in Assessment of Rice Blast Resistance [J]. Rice Science, 2023, 30(6): 10-.
[7]	Md. Forshed DEWAN, Md. AHIDUZZAMAN, Md. Nahidul ISLAM, Habibul Bari SHOZIB. Potential Benefits of Bioactive Compounds of Traditional Rice Grown in South and South-East Asia: A Review [J]. Rice Science, 2023, 30(6): 5-.
[8]	Raja CHAKRABORTY, Pratap KALITA, Saikat SEN. Phenolic Profile, Antioxidant, Antihyperlipidemic and Cardiac Risk Preventive Effect of Chakhao Poireiton (A Pigmented Black Rice) in High-Fat High-Sugar induced Rats [J]. Rice Science, 2023, 30(6): 11-.
[9]	LI Qianlong, FENG Qi, WANG Heqin, KANG Yunhai, ZHANG Conghe, DU Ming, ZHANG Yunhu, WANG Hui, CHEN Jinjie, HAN Bin, FANG Yu, WANG Ahong. Genome-Wide Dissection of Quan 9311A Breeding Process and Application Advantages [J]. Rice Science, 2023, 30(6): 7-.
[10]	JI Dongling, XIAO Wenhui, SUN Zhiwei, LIU Lijun, GU Junfei, ZHANG Hao, Tom Matthew HARRISON, LIU Ke, WANG Zhiqin, WANG Weilu, YANG Jianchang. Translocation and Distribution of Carbon-Nitrogen in Relation to Rice Yield and Grain Quality as Affected by High Temperature at Early Panicle Initiation Stage [J]. Rice Science, 2023, 30(6): 12-.
[11]	Nazaratul Ashifa Abdullah Salim, Norlida Mat Daud, Julieta Griboff, Abdul Rahim Harun. Elemental Assessments in Paddy Soil for Geographical Traceability of Rice from Peninsular Malaysia [J]. Rice Science, 2023, 30(5): 486-498.
[12]	Monica Ruffini Castiglione, Stefania Bottega, Carlo Sorce, Carmelina SpanÒ. Effects of Zinc Oxide Particles with Different Sizes on Root Development in Oryza sativa [J]. Rice Science, 2023, 30(5): 449-458.
[13]	Tan Jingyi, Zhang Xiaobo, Shang Huihui, Li Panpan, Wang Zhonghao, Liao Xinwei, Xu Xia, Yang Shihua, Gong Junyi, Wu Jianli. ORYZA SATIVA SPOTTED-LEAF 41 (OsSPL41) Negatively Regulates Plant Immunity in Rice [J]. Rice Science, 2023, 30(5): 426-436.
[14]	Ammara Latif, Sun Ying, Pu Cuixia, Noman Ali. Rice Curled Its Leaves Either Adaxially or Abaxially to Combat Drought Stress [J]. Rice Science, 2023, 30(5): 405-416.
[15]	Liu Qiao, Qiu Linlin, Hua Yangguang, Li Jing, Pang Bo, Zhai Yufeng, Wang Dekai. LHD3 Encoding a J-Domain Protein Controls Heading Date in Rice [J]. Rice Science, 2023, 30(5): 437-448.