Agrobacterium rhizogenes K599-mediated chrysanthemum transgenic method

Abstract

The invention provides an agrobacterium rhizogenes K599-mediated chrysanthemum transgenic method, which has the high regeneration efficiency of chrysanthemum. In the method, a living plant material is used as a receptor-induced adventitious root (lamina wounds or axillae of the chrysanthemum) infected by agrobacterium rhizogenes K599 directly, and callus is induced by the adventitious root to undergo differentiation culturing to grow into an integral plant. The method has the advantages that false positive adventitious roots can be prevented; simultaneously, based on the characteristic of sterile growing point regions at the top end in the multiplicative process of the adventitious root, the aims of inhibiting and killing agrobacterium can be fulfilled by cutting the regions of growing points adjacent to the top end of the adventitious root for subculturing and by combining sterilization by cephalosporin; and sterilization by antibiotics in the processes of induced healing and differentiation of the adventitious root is avoided, so the transgenic regeneration efficiency is improved.

Description

(1) Technical field [0001] The invention relates to a chrysanthemum transgenic method mediated by Agrobacterium rhizogenes K599. (2) Background technology [0002] Chrysanthemum (Dendranthema morifolium) belongs to the plant of Compositae Chrysanthemum. It is native to my country. It is one of the top ten traditional famous flowers in my country and one of the four major cut flowers in the world. It is also a famous flower integrating cultural significance and economic value. It has edible and medicinal properties. Use value (Dai Silan et al., Chrysanthemum Systematics and Chrysanthemum Origin Research Progress, Journal of Beijing Forestry University, 2002, 24 (5 / 6): 230-234). Plant shape, flower type and flower color are important factors affecting the quality of flowers such as chrysanthemums, and plant transgenic technology plays an important role in improving flower quality. Shao Hanshuang et al., Petty et al., Zheng et al. Genes such as Phy B-1 were transferred into chr...

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Problems solved by technology

In addition, it has been reported that using the Agrobacterium transgenic method to transfer target genes with different functions into chrysanthemums (Fu Rongzhao and Liu Min, Obtaining chrysanthemum transgenic plants through the Agrobacterium tumefaciens-mediated method, Journal of Plant Physiology, 1998, 24 (1): 72-76; Wang Guanlin et al., Studies on aphid resistance of chrysanthemums transformed with snowdrop lectin gene and transgenic plants, Acta Genetica Sinica, 2004, 31(12): 1434-1438; Hong Bo et al., Increased heterologous expression of AtDREB1A gene in chrysanthemums Tolerance of plants to drought and salinity stress, Chinese Science (Series C), 2006, 36(3): 223-231; Hong Bo et al., Regeneration of Somatic Embryo Pathway of Ground Cover Chrysanthemum Fall Color, Genetic Transformation and Resistance of Transgenic Plants Detection of coldness, Chinese Agricultural Sciences, 2006, 39(7): 1443-1450; Sun Lei et al., Obtaining non-selectable marker transgenic chrysanthemums using double T-DNA vector system, Acta Horticultural Science, 2008, 35(5): 727-734 , Jiang Dan et al. Transformation of cut chrysanthemum 'Shenma' with Arabidopsis thaliana flowering gene FT, Acta Horticultural Sinica, 2010, 37(3): 423-430), but the regeneration efficiency of chrysanthemum in the above reports is low

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