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Creation of Transgenic Bananas Expressing Human Lysozyme Gene for Panama Wilt Resistance
Authors:Xin-Wu PEI  Shi-Kai CHEN  Rui-Ming WEN  Shang YE  Jia-Qin HUANG  Yong-Qiang ZHANG  Bing-Shan WANG  Zhi-Xing WANG  Shi-Rong JIA
Institution:Biotechnology Institute, Chinese Academy of Agricultural Sciences;, Beijing 100081, China; Xinyi Institute of Agricultural Science and Technology;, Xinyi, Guangdong 523530, China
Abstract:Human lysozyme (HL) inhibits Fusarium oxysporum (FocR4) growth in vitro. To obtaintransgenic bananas (Musa spp.) that are resistant to Panama wilt (F. oxysporum), we introduced an HL genethat is driven by a constitutive cauliflower mosaic virus 35S promoter into the banana via Agrobacterium-mediated transformation. PCR confirmed that 51 transgenic plants were obtained. The development ofPanama wilt symptoms were examined after the plants had been grown in pots. The non-transgenic plantsdeveloped typical fusarium symptoms 60 d after FocR4 inoculation, whereas 24 of 51 transgenic plants remained healthy. The transgenic banana plants that showed resistance to FocR4 in the pots were then planted in a field that was heavily infected with FocR4 for further investigation. Eleven of 24 plants developed symptoms before bud emergence; another 11 plants showed symptoms after bud emergence and the remaining two plants, H-67 and H-144, remained healthy and were able to fruit. Northern blotting analysisdemonstrated that H-67 and H-144, bearing the strongest resistance to Panama wilt, had the highest level ofHL expression and that the expression of HL was well correlated with the FocR4 resistance of transgenicplants. We conclude that Agrobacterium-mediated transformation, with the assistance of particlebombardment, is a powerful approach for banana transformation and that a transgenic HL gene can causeresistance of the crop to FocR4 in the field.
Keywords:Agrobacterium-mediated transformation  Fusarium oxysporum  human lysozyme  Panamawilt disease  particle bombardment  transgenic banana
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