Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (2): 215-228.doi: 10.3864/j.issn.0578-1752.2015.02.02

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Production of Transgenic Maize Germplasm with Multi-Traits of Insect-resistance, Glyphosate-Resistance and Drought-Tolerance

SUN Yue1, LIU Xiu-xia1, LI Li-li2, GUAN Yun-yun1, ZHANG Ju-ren1   

  1. 1School of Life Science, Shandong University, Jinan 250100
    2Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan 250100
  • Received:2014-06-11 Online:2015-01-16 Published:2015-01-16

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Abstract:

【Objective】 The objective of this experiment is to study the production of transgenic maize with triple resistance of insect, glyphosate and drought stress. 【Method】In this study, an expression vector harboring four target genes (cry1AcM, epsps, GAT and ZmPIS）was used to transform to maize elite inbred lines 9801 and Qi 319 (Q319) mediated by Agrobacterium, and produced transgenic maize plants with multiple modified traits. Based on herbicide resistance, PCR analysis, transcriptional analysis, western blot detection and insect resistance assay, six stable transgenic maize lines were selected from a large number of transgenic lines. Subsequently, the inbred lines 9801 and Q319 were used as the nontransgenic control to analyze the resistance of transgenic plants to Ostrinia furnacalis In view of different resistances to O. furnacalis of maize plants at different developmental stages, the resistance to O. furnacalis of transgenic plants at different developmental periods was tested indoor and in the fields by innoculating the worm of O. furnacalis, and kernels and the bracts of ears from transgenic plants at filling stage were used to feed the worm. In the herbicide resistance trials in fields, transgenic plants at the six-leaf stage were sprayed with 0.84 kg·hm-2 glyphosate (commercial application dosage) to evaluate the resistance of transgenic plants. A spraying experiment with triple dose of glyphosate on the plants at three-leaf stage was also performed to evaluate the glyphosate-resistant levels of transgenic lines. To evaluate the resistance of drought stress, the morphological changes of transgenic plants were observed and their photosynthesis and chlorophyll fluorescence were measured on plants at the 10-leaf stage in a drought stress treatment. 【Result】Among six stable transgenic maize lines, L1-L3 come from inbred line 9801 and line Q1-Q3 come from inbred line Q319. The stable expression of the four transgenic genes in the transgenic lines was confirmed by using RT-PCR assay. The levels of cry1Ac protein in the different transgenic lines were determined by Western blot, which showed reliable expression in various organs of the transgenic lines. Six lines with higher resistance to the corn borer compared with wild type were selected through the corn borer resistance experiment in the plants at vegetative stage and filling stage. For herbicide resistance trials, the glyphosate resistance of transgenic plants showed more tolerant than that of wild type control. During drought stress, the transgenic plants had more strong photosynthetic capacity and photosystem II activity than that of the nontransgenic control plants under drought stress conditions. 【Conclusion】The introduction of the cry1Ac-M, epsps, GAT, and ZmPIS genes into maize elite inbred lines increased the insect-resistance, glyphosate-tolerance, and improved the drought-resistance of transgenic plants, and the transgenic plant are up to the level of extensive utilization in corn production. Six new corn transgenic lines with excellent complex traits were obtained in this study., glyphosate and drought under strictly controlled experimental conditions.

Key words: resistance genetically modified maize, complex traits; insect resistance, herbicide tolerance, drought

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