水稻规模化转基因技术体系构建与应用

doi:10.3864/j.issn.0578-1752.2014.21.002

Abstract

Abstract: Rice, one of the most important food crops and a model monocot plant, its genetic transformation has attracted widespread attention. Since the world’s first case of transgenic rice got success in 1988, the rice genetic transformation technology has been developed rapidly. In 1994, in a milestone work of Agrobacterium-mediated transformation of a number of japonica rice cultivars, rice genetic transformation system has been more advanced after nearly 20 years of development. Presently, Agrobacterium-mediated transformation and microparticle bombardment are the two most widely used methods for rice genetic transformation, while pollen tube pathway, electroporation and polyethylene glycol (PEG) medium are also employed in some laboratories. Agrobacterium transformation is the method of choice because it is of easy and low cost, the high transformation efficiency, the low copy number of transgenes inserted into the host genome and the stability of expression over generations. Nowadays more than 80% of the transgenic rice is transformed by the Agrobacterium method. Although there are a lot of reports in improving the methods and technology of rice transformation, difficulties and limitations still exist in rice genetic transformation. The transformation efficiency of several elite japonica and many indica varieties is still unsatisfactory as it is much lower. Some transformation protocols are laborious, time consuming, and highly genotype-dependent. Besides, there is a problem of somaclonal variations or somatic mutation frequently occurs in plant cells during in vitro culture. Therefore, it is very important to establish the high efficient, safety, large-scale and standard transformation systems for diverse varieties of rice. This paper briefly reviewed the history and development of rice genetic transformation system. Especially, the recent progress of rice transgenic technology for large scale performance in China was described. Many factors are known to affect the efficiency of genetic transformation, such as the genotype of the recipient plants, the type and age of the tissue being inoculated, the strain of Agrobacterium, the expression vector, the composition of the culture medium, as well as various conditions of tissue culture. Extensive research has been conducted to optimize the high-efficiency transformation system for different genotypes of rice. Various factors affecting the transformation efficiencies were explored. Modification in culture conditions for embryogenic calli induction and regeneration from mature seed and also the compatibility of the Agrobacterium strain were successful in examined transformation protocols. The highly efficient Agrobacterium-mediated transformation system for japonica and indica rice using immature embryos or mature embryo-derived calli as the explant has been established for the large-scale production of transgenic rice by integrating new technology method. Also the Agrobacterium-mediated co-transformation system has been preliminary developed and some experimental parameters for marker-free rice transformation have been optimized. However, a big gap between the update status in China and the advanced level in some renowned international biotechnology companies is still existed in either transformation efficiency or transformation scale of rice. It was concluded that bio-safety, high efficiency and large scale were the bottlenecks for production GM rice products and its industrialization. Future studies in this field should be focused on the use of commercial rice varieties, improvement of transformation efficiency, development of marker-free rice plants, introduction of multiple target genes, integration of foreign genes at specific site, etc. The review also analyzed the problems existing in large-scale rice transformation system and provided strategies to improve it. It may provide some useful information for efficient production of transgenic rice for practical use as well as for studying gene function.

Key words: rice, largescale, transformation system

ZHANG Xin, FU Ya-ping, ZHOU Jun-li, GUO Xiu-ping, LIU Wen-zhen, WU Jie-fang, WU Chuan-yin, WAN Jian-min. Establishment and Application of Large-Scale Transformation Systems for Rice[J].Scientia Agricultura Sinica, 2014, 47(21): 4141-4154.

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Establishment and Application of Large-Scale Transformation Systems for Rice

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