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The present study was conducted to determine the effect of exogenous application of brassinolide (BR) on Leymus chinensis grown under shade, i.e., control (100% natural light), mild shade (70% natural light), and moderate shade (50% natural light). Shade substantially enhanced the plant growth, synthesis of photosynthetic pigments, photosynthetic efficiency, and chlorophyll (Chl) fluorescence attributes of L. chinensis as compared with control. The order of increase was mild shade > moderate shade > natural light except Chl content, where the order of increase was moderate shade > mild shade > natural light. Likewise, application of BR resulted in further exacerbation of plant height, plant fresh and dry mass, but less in case of Chl and carotenoids contents, gas-exchange characteristics, and Chl fluorescence attributes. The results conclude that shade significantly enhanced plant growth through alterations in physiological attributes of L. chinensis, while, application of BR may not further improve the plant growth under shade.

This study aimed to develop a new vector system to remove selection genes and to introduce two or more genes of interest into plants in order to express them in a coordinated manner. A multigene expression vector was established based on pCamBIA2300 using a selectable marker gene (SMG)-free system based on the combination of the isocaudamer technique and double T-DNA. The vector DT7 containing seven target genes was constructed and introduced into tobacco using Agrobacterium-mediated transformation. Twenty-one of 27 positive transgenic plants contained both T-DNA regions. The co-transformation frequency was 77.8 %. The frequency of unlinked integration of two intact T-DNAs was 22.22 % (6/27). The frequency of removal of SMG from transgenic T1 plants was 19.10 %. These results suggest that this vector system was functional and effective for multigene expression and SMG-free transgenic plant cultivation. At least seven target genes can be co-expressed using this system. Overall, these findings provide a new and highly effective platform for multigene and marker-free transgenic plant production.