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| The Mat# Gene Optimized by Codon Bias of Rice (Oryza sativa) Enhances Resistance to Glufosinate |
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Abstract Rice (Oryza sativa) is one of the most important food crops in the world. Among the factors harmful to rice production, weeds results in the most severe yield loss. To control weeds by herbicide is convenience, fast and low cost, that becomes the first choice in weeding. To cultivate transgenic non-selective herbicide resistant crops can decrease the kind, dosage and residual of herbicide, and prompt the productive efficiency of land, that plays an important role in commercial crop production. The Mat (methionine sulfone N-acetyltransferase) gene from bacterium Nocardia sp. AB2253 is a novel glufosinate resistant gene reported in 2009, which may be useful to develop herbicide resistant crops. However, the previous studies showed that the original Mat gene was not powerful enough in transgenic rice to provide high level of glufosinate resistance. In this study, the optimized Mat# gene according to codon bias in rice was transformed into an indica rice cultivar 9K (O. sativa ssp. indica) by use of Agrobacterium-mediated method. PCR analysis and Southern blot analysis of transgenic plants showed that the target gene Mat# had been integrated into rice genome with two copies. Glufosinate spraying on T2 generation plants at seedling stage showed that the T2 generation plants were resistant to glufosinate with a tolerance concentration of 1 000 mg/L at least. Progeny segregation statistics showed that the ratio of plant resistant and sensitive to glufosinate in T2 generation was 3.56∶1, which was conform to Mendelian inheritance by Chi-square test. Germination of T3 generation seeds under glufosinate stress showed that the T3 generation plants were resistant to glufosinate with a tolerance concentration of 600 mg/L at least, which indicated the resistance level of transgenic plants with Mat# gene was not lower than that of 9KA2 with Bar gene transformed. The investigation of agronomic traits showed that there was no significant differences in main agronomic traits between the T2 generation plants and non-transgenic controls, which indicated the introduction of alien gene did not have an obvious impact on main agronomic traits. Enzyme activity assay showed that the average activity of N-acetyltransferase in leaf was about 6.6 times of that in non-transgenic control. These results suggestted that the optimized Mat# gene provided higher glufosinate resistance than the original version, which can be used as a selection marker and new herbicide resistant gene resource in transgenic crop breeding.
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Received: 29 April 2016
Published: 06 August 2016
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