THE EFFECTS OF ANTIOXIDANTS ON THE TRANSIENT EXPRESSION OF GUS GENE IN SOYBEAN HYPOCOTYLS MEDIATED BY AGROBACTERIUM TUMEFACIENS

doi:10.17521/cjpe.2006.0044

Abstract

Abstract:

Soybean hypocotyls have advantages of higher regeneration frequency and faster formation of adventitious shoots, and often be used in soybean transformation mediated by Agrobacterium tumefaciens. But the browning and necrosis of hypocotyls during co_cultivation greatly affects the efficiency of the transformation.The effects of antioxidants on the transient expression efficiency of GUS (β_glucuronidase) gene in soybean hypocotyls mediated byAgrobacterium tumefaciens were studied by adding antioxidants (thiosulfate, L_cysteine, and dithiothreitol) in the co_cultivation medium,and the transient expression efficiency of the GUS gene was measured by histochemically method. The results showed that the antioxidant in the co_cultivation medium increased the transient expression efficiency up to 23.4% in hypocotyls. Further investigation demonstrated that higher transient expression efficiency was associated with concomitant decreases in the levels of hypocotyl browning. The stable expression of GUS gene in hypocotyls was found in the cambium cells, which was formed as deep blue zone on the hypocotyls. Our results also showed that concentration of Agrobacterium tumefaciens and co_cultivation time were the important factors that affected the transient expression efficiency of GUS gene in hypocotyls. In our experiments, the highest transient expression efficiency of GUS gene obtained by inoculating hypocotyls in Agrobacterium tumefaciens (OD₆₀₀ 0.6) and co_cultivating for 3 days in the darkness. All results suggested that antioxidants could efficiently improve transient expression efficiency of GUS gene in soybean hypocotyls.

Key words: Soybean, Hypocotyls, Agrobacterium tumefaciens, GUS gene, Transient expression

JI Feng_Yuan, WANG Ge_Liang, XU Yi_Nong. THE EFFECTS OF ANTIOXIDANTS ON THE TRANSIENT EXPRESSION OF GUS GENE IN SOYBEAN HYPOCOTYLS MEDIATED BY AGROBACTERIUM TUMEFACIENS[J]. Chin J Plant Ecol, 2006, 30(2): 330-334.

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URL: https://www.plant-ecology.com/EN/10.17521/cjpe.2006.0044

https://www.plant-ecology.com/EN/Y2006/V30/I2/330

Figures/Tables 5

Fig.1 Transient expression efficiency of GUS gene in soybean hypocotyls

Fig.2 The effects of antioxidants on the browning, necrosis of hypocotyls and the location of GUS expression in soybean hypocotyls

Fig.3 The effects of co_cultivation time on transient expression efficiency of GUS gene in soybean hypocotyls

Fig.4 The effects of re_suspend liquid medium dilute consistence on GUS gene transient expression efficiency in soybean hypocotyls

Fig.5 The effects of different soybean species on soybean hypocotyls GUS gene transient expression efficiency

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