Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana....Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana. Whether TCP1 can directly bind to the promoter region of DWF4, however, has not been experimentally demonstrated. Here we provide our biochemical and genetic evidence that TCP1 mediates the expression of DWF4 by directly associating with the two GGNCCC motifs in the promoter region of DWF4. The expression levels of DWF4 are positively correlated to TCP1 abundance in planta. Electrophoretic mobility shift assays （EMSAs） using various synthetic DNA fragments suggest that the GGNCCC core sequence is critical for TCP1 binding. DNA sequences flanking the GGNCCC motifs are less important for the association of TCPI. Using DWF4p-GUS transgenic plants as an assay system, it is clearly indicated that these motifs are required for the positive regulation of DWF4 transcription by TCP1. More significantly, whole genome microarray analyses indicate that TCP1 can directly or indirectly regulate the expression of many other genes known to be important for normal plant growth and development.展开更多
Brassinosteroids（BRs）, a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable ...Brassinosteroids（BRs）, a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable and has been adopted as the model species for studying fruit growth, development, and ripening. To understand the role of endogenous BRs in the development of tomato fruit, the expression patterns of three homologues of DWF4 gene were investigated and the transgenic tomato plants were generated in which the Gh DWF4 gene from upland cotton（Gossypium hirsutum L.） was ectopically expressed. The contents of main quality components were analyzed in fruits of transgenic tomato line and non-transgenic line（control plant, CP） when the fruit was mature. Sl CYP90B3 that possesses high homology with Gh DWF4 preferentially expressed in mature fruit. Significantly higher contents of soluble sugar, soluble proteins, and vitamin C were obtained in fruit of transgenic tomato lines compared with those in the CP. Furthermore, overexpressing Gh DWF4 promoted fruit growth and ripening. The weight per fruit was increased by about 23% in transgenic lines. In addition, overexpressing Gh DWF4 promoted the germination of transgenic tomato seeds and hypocotyl elongation of seedlings. These results indicated that overexpressing Gh DWF4 gene in tomato could increase the contents of many nutrients in fruit and accelerate fruit ripening. It is suggested that increased endogenous BRs in fruit affect the growth and development of tomato fruit and therefore improved the nutrient quality of tomato.展开更多
基金supported by the National Basic Research Program of China(Grant No.2011CB915401)the National Natural Science Foundation of China(Grant Nos. 90917019,91117008 and 91317311)
文摘Our previous studies indicated that TCP1 is a positive regulator of the brassinosteroid （BR） biosynthesis pathway by mediating the transcription of DWF4, one of the key BR biosynthetic genes in Arabidopsis thaliana. Whether TCP1 can directly bind to the promoter region of DWF4, however, has not been experimentally demonstrated. Here we provide our biochemical and genetic evidence that TCP1 mediates the expression of DWF4 by directly associating with the two GGNCCC motifs in the promoter region of DWF4. The expression levels of DWF4 are positively correlated to TCP1 abundance in planta. Electrophoretic mobility shift assays （EMSAs） using various synthetic DNA fragments suggest that the GGNCCC core sequence is critical for TCP1 binding. DNA sequences flanking the GGNCCC motifs are less important for the association of TCPI. Using DWF4p-GUS transgenic plants as an assay system, it is clearly indicated that these motifs are required for the positive regulation of DWF4 transcription by TCP1. More significantly, whole genome microarray analyses indicate that TCP1 can directly or indirectly regulate the expression of many other genes known to be important for normal plant growth and development.
基金supported by the Natural Science Foundation of Chongqing, China (CSTC, 2011BB1007)the Genetically Modified Organisms Breeding Major Projects of China (2009ZX08009-118B)
文摘Brassinosteroids（BRs）, a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable and has been adopted as the model species for studying fruit growth, development, and ripening. To understand the role of endogenous BRs in the development of tomato fruit, the expression patterns of three homologues of DWF4 gene were investigated and the transgenic tomato plants were generated in which the Gh DWF4 gene from upland cotton（Gossypium hirsutum L.） was ectopically expressed. The contents of main quality components were analyzed in fruits of transgenic tomato line and non-transgenic line（control plant, CP） when the fruit was mature. Sl CYP90B3 that possesses high homology with Gh DWF4 preferentially expressed in mature fruit. Significantly higher contents of soluble sugar, soluble proteins, and vitamin C were obtained in fruit of transgenic tomato lines compared with those in the CP. Furthermore, overexpressing Gh DWF4 promoted fruit growth and ripening. The weight per fruit was increased by about 23% in transgenic lines. In addition, overexpressing Gh DWF4 promoted the germination of transgenic tomato seeds and hypocotyl elongation of seedlings. These results indicated that overexpressing Gh DWF4 gene in tomato could increase the contents of many nutrients in fruit and accelerate fruit ripening. It is suggested that increased endogenous BRs in fruit affect the growth and development of tomato fruit and therefore improved the nutrient quality of tomato.
