In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vec...In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vector pSB130, respectively, and introduced into the calli derived from the immature seeds of two elite japonica rice varieties, Guangling Xiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many cotransgenic rice lines containing both the AP1 gene and the marker gene were regenerated and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable marker-free transgenic rice plants were subsequently obtained from the progeny of the cotransformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.展开更多
This study was conducted to investigate changes in the expression of AP1 gene in flowering process. Potassium nitrate and ethephon were sprayed on 7- year-old Guifei trees out of season. The results showed that AP1 ge...This study was conducted to investigate changes in the expression of AP1 gene in flowering process. Potassium nitrate and ethephon were sprayed on 7- year-old Guifei trees out of season. The results showed that AP1 gene had a higher expression level in terminal buds, and especially, the expression level increased significantly in late stage of flower bud differentiation. Potassium nitrate and ethephon promoted flower bud differentiation, and the expression level of AP1 gene in- creased in flowering process remarkably. Expression ofAP1 gene of the potassium nitrate treatment was significantly greater than that of the ethephon treatment and the CK.展开更多
The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were st...The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were studied.The results showed that the leaf explants precultured for 2～8 hours or not precultured were best for transformation by A. tumefaciens. The suitable concentration of bacterial and the time for infecting was OD_ 600 0.5 for 10 minutes. Leaves were cocultivated with bacterial at 23～25 ℃ for 2 days,then delayed selection for 3 days. Kan^r plant were selected by increased selective press from 5 mg·L ~ -1 G418 to 7.5 mg5L~ -1 G418 followed by 10 mg5L~ -1 G418. The integration of AP1 gene into C. morifolium ‘Yu Ren Mian’ was confirmed by PCR and Southern blotting.Two of transgenic plants bloomed 15 days earlier than untransformed plants.展开更多
基金This paper is translated from its Chinese version in Scientia Agricultura Sinica.This study was supported by the Government of Jiangsu Province,China(BG2002301 and JH02-106)National Transgenic Plant R&D Project(JY03-B-10)+1 种基金National Natural Science Foundation of China(30170567)Department of Education of Jiangsu Goverment,China(K05015).
文摘In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vector pSB130, respectively, and introduced into the calli derived from the immature seeds of two elite japonica rice varieties, Guangling Xiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many cotransgenic rice lines containing both the AP1 gene and the marker gene were regenerated and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable marker-free transgenic rice plants were subsequently obtained from the progeny of the cotransformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.
基金Supported by CATAS-TCGRI(1630032013010)Special Fund for Agro-scientific Research in the Public Interest(201203092)
文摘This study was conducted to investigate changes in the expression of AP1 gene in flowering process. Potassium nitrate and ethephon were sprayed on 7- year-old Guifei trees out of season. The results showed that AP1 gene had a higher expression level in terminal buds, and especially, the expression level increased significantly in late stage of flower bud differentiation. Potassium nitrate and ethephon promoted flower bud differentiation, and the expression level of AP1 gene in- creased in flowering process remarkably. Expression ofAP1 gene of the potassium nitrate treatment was significantly greater than that of the ethephon treatment and the CK.
文摘The flower’s meristematic characteristic gene AP1 was introduced into Chrysanthemum morifolium cv. ‘Yu Ren Mian’ mediated by Agrobacterium tumefaciens.The factors influencing genetic transformation protocol were studied.The results showed that the leaf explants precultured for 2～8 hours or not precultured were best for transformation by A. tumefaciens. The suitable concentration of bacterial and the time for infecting was OD_ 600 0.5 for 10 minutes. Leaves were cocultivated with bacterial at 23～25 ℃ for 2 days,then delayed selection for 3 days. Kan^r plant were selected by increased selective press from 5 mg·L ~ -1 G418 to 7.5 mg5L~ -1 G418 followed by 10 mg5L~ -1 G418. The integration of AP1 gene into C. morifolium ‘Yu Ren Mian’ was confirmed by PCR and Southern blotting.Two of transgenic plants bloomed 15 days earlier than untransformed plants.
