Many stress responsive genes have been reported with an effect on improving stress resistance in model plants under greenhouse conditions. Towards identification of genes for drought resistance breeding, seven well do...Many stress responsive genes have been reported with an effect on improving stress resistance in model plants under greenhouse conditions. Towards identification of genes for drought resistance breeding, seven well documented genes （CBF3, SOS2, NCED2, NPK1, LOSS, ZAT10, and NHX1） in stress resistance were selected in this study and transformed into rice cultivar Zhonghua 11 under the control of constitutive promoter Actinl and stress-inducible pro- moter of a rice HVA22 homolog, and transgenic rice were tested for drought resistance under field conditions. A total of 1598 independent transgenic To plants were generated. The percentages of single copy and expression of the transgenes were 36.7% and 57.6%, respectively. For each gene construct, 30 T1 families with expression of transgene were selected for drought resistance testing at the reproductive stage in field, and 10 of them were tested in PVC pipes with a defined stress protocol at the same stage. Relative yield and relative spikelet fertility were used as two major criteria to evaluate drought resistance performance because significantly decreased yield was observed in the T1 generation, Trans- genic families of eight constructs （HVA22P：CBF3, HVA22P：NPK1, Actin 1：LOS5, HVA22P：L OS5, Actin 1：ZA T10, HVA22P：ZA T10, Actinl：NHX1, and HVA22P：NHX1） showed significantly higher RY than wild-type （WT） under both drought stress field and PVC tube conditions. Transgenic families of 9 constructs （HVA22P.SOS2 and CBF3, LOS5, ZAT10, and NHX1 by both promoters） showed significantly higher relative spikelet fertility than WT in the field or PVC pipes. In the field drought resistance testing of T2 families derived from the T1 families with relatively lower yield decrease, transgenic families of seven constructs （HVA22P：CBF3, Actinl：NPK1, HVA22P：NPK1, Actinl：LOS5, HVA22P：LOS5, Actin1：ZAT10, and HVA22P：ZAT10） showed significantly higher yield per plant than WT, and families of nine constructs （Actinl：CBF3, HVA22P：CBF3, HVA22P：- SOS2, HVA22P：NPK1, Actinl：LOS5, HVA22P：LOS5, Actin1：ZAT10, HVA22P：ZAT10, and Actinl：NHX1） had higher spikelet fertility than WT. In general, LOS5 and ZAT10 showed relatively better effect than the other five genes in improving drought resistance of transgenic rice under field conditions. The results and experience obtained from this study could be a useful reference for drought resistance engineering in rice.展开更多
A series of high-throughput binary cloning vectors were constructed to facilitate gene function analysis in higher plants. This vector series consists of plasmids designed for plant expression, promoter analysis, gene...A series of high-throughput binary cloning vectors were constructed to facilitate gene function analysis in higher plants. This vector series consists of plasmids designed for plant expression, promoter analysis, gene silencing, and green fluorescent protein fusions for protein localization. These vectors provide for high-throughput and efficient cloning utilizing sites for λ phage integrase/excisionase. In addition, unique restriction sites are incorporated in a multiple cloning site and enable promoter replacement. The entire vector series are available with complete sequence information and detailed annotations and are freely distributed to the scientific community for non-commercial uses.展开更多
文摘Many stress responsive genes have been reported with an effect on improving stress resistance in model plants under greenhouse conditions. Towards identification of genes for drought resistance breeding, seven well documented genes （CBF3, SOS2, NCED2, NPK1, LOSS, ZAT10, and NHX1） in stress resistance were selected in this study and transformed into rice cultivar Zhonghua 11 under the control of constitutive promoter Actinl and stress-inducible pro- moter of a rice HVA22 homolog, and transgenic rice were tested for drought resistance under field conditions. A total of 1598 independent transgenic To plants were generated. The percentages of single copy and expression of the transgenes were 36.7% and 57.6%, respectively. For each gene construct, 30 T1 families with expression of transgene were selected for drought resistance testing at the reproductive stage in field, and 10 of them were tested in PVC pipes with a defined stress protocol at the same stage. Relative yield and relative spikelet fertility were used as two major criteria to evaluate drought resistance performance because significantly decreased yield was observed in the T1 generation, Trans- genic families of eight constructs （HVA22P：CBF3, HVA22P：NPK1, Actin 1：LOS5, HVA22P：L OS5, Actin 1：ZA T10, HVA22P：ZA T10, Actinl：NHX1, and HVA22P：NHX1） showed significantly higher RY than wild-type （WT） under both drought stress field and PVC tube conditions. Transgenic families of 9 constructs （HVA22P.SOS2 and CBF3, LOS5, ZAT10, and NHX1 by both promoters） showed significantly higher relative spikelet fertility than WT in the field or PVC pipes. In the field drought resistance testing of T2 families derived from the T1 families with relatively lower yield decrease, transgenic families of seven constructs （HVA22P：CBF3, Actinl：NPK1, HVA22P：NPK1, Actinl：LOS5, HVA22P：LOS5, Actin1：ZAT10, and HVA22P：ZAT10） showed significantly higher yield per plant than WT, and families of nine constructs （Actinl：CBF3, HVA22P：CBF3, HVA22P：- SOS2, HVA22P：NPK1, Actinl：LOS5, HVA22P：LOS5, Actin1：ZAT10, HVA22P：ZAT10, and Actinl：NHX1） had higher spikelet fertility than WT. In general, LOS5 and ZAT10 showed relatively better effect than the other five genes in improving drought resistance of transgenic rice under field conditions. The results and experience obtained from this study could be a useful reference for drought resistance engineering in rice.
文摘A series of high-throughput binary cloning vectors were constructed to facilitate gene function analysis in higher plants. This vector series consists of plasmids designed for plant expression, promoter analysis, gene silencing, and green fluorescent protein fusions for protein localization. These vectors provide for high-throughput and efficient cloning utilizing sites for λ phage integrase/excisionase. In addition, unique restriction sites are incorporated in a multiple cloning site and enable promoter replacement. The entire vector series are available with complete sequence information and detailed annotations and are freely distributed to the scientific community for non-commercial uses.
