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| Establishment of an Optimized Co-transformation System of Common Wheat (Triticum aestivum) Using Particle Bombardment |
| HE Xiao-Lan1,2, WANG Jian-Wei1, CHEN Xin-Hong2,*, LI Wen-Xu3, TANG Hong1 |
1 School of Life and Health Science, Kaili University, Kaili 556011, China;
2 Shaanxi Key Laboratory of Genetic Engineering for Plant Breeding/College of Agronomy, Northwest A&F University, Yangling 712100, China;
3 Institute for Wheat Research, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China |
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Abstract Along with the increased commercialization and planting of a range of genetically modified (GM) crops globally, the biosafety of GM crops has become a hot topic and major public concern. To elucidate the optimal molar ratio for co-transformation and develop a safe, high-efficiency wheat (Triticum aestivum) transformation system, in this study, a combination of β-glucuronidase (gus) and a herbicide resistance gene (bar) was used to select stable transformants. Six independent co-transformation experiments were performed using either the minimal cassette Ubi-bar or the whole of pAHC20, with the Ubi-gus minimal cassette at different molar ratios. The whole pAHC25 plasmid was used as a reference, which was transferred into immature embryos of the wheat variety 'Kenong 199'. The results showed that the transient expression rate of gus gene increased significantly with the increase of Ubi-gus molar concentration, but there was no correlation between transient expression and stable expression. The transformation frequency differed significantly among combinations, in which the highest was obtained by combining Ubi-bar or pAHC20∶Ubi-gus at a molar ratio of 1∶2. The transformation frequency was significantly lower using the whole plasmid than the minimal constructs with co-transformation at a molar ratio of 1∶2, but significantly higher than that with 1∶1 and 1∶3. The stable integration, expression, and inheritance of the gus were confirmed by GUS assays and Southern blotting. The positive transgenic lines exhibited normal morphology except for poor seed set. This system may facilitate future transformation studies.
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Received: 28 May 2020
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Corresponding Authors:
*cxh2089@126.com
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