Oryza sativa L. ssp. japonica and indica exhibit different sensitivity to photoinhibition and they show different stability of their core proteins D1 in the chloroplast photosystem Ⅱ. Using in situ hybridization, psb...Oryza sativa L. ssp. japonica and indica exhibit different sensitivity to photoinhibition and they show different stability of their core proteins D1 in the chloroplast photosystem Ⅱ. Using in situ hybridization, psbA, the gene encoding D1 protein of O. sativa ssp. japonica cv. 9516, and that of O. sativa ssp. indica cv. Shanyou 63 was cloned. As revealed by homology comparison of their sequences, the sequences are identical in the regions of promoter and 5′-UTR; differences are found in individual bases in the coding region all of which, being in the third position of respective codons, however, do not affect the amino acids coded finally; a difference is noted in the length of the oligo-U sequence in the region of 3′-UTR. It is thus apparent that, rather than a result of any difference in the amino acid sequences, the differences in the sensitivity to photoinhibition of D1 proteins between japonica and indica rice may be related to the upstream factors that regulate expression of psbA or to differences of photoprotective mechanisms.展开更多
钙离子作为植物细胞的第二信使,广泛参与植物应对不同逆境胁迫的信号调控过程。水稻G蛋白促进蛋白1(Oryza sativa GTPase-activating protein 1,OsGAP1)包含1个C2结构域,而含C2结构域的蛋白质是一类钙离子结合蛋白质,受钙信号的调控。...钙离子作为植物细胞的第二信使,广泛参与植物应对不同逆境胁迫的信号调控过程。水稻G蛋白促进蛋白1(Oryza sativa GTPase-activating protein 1,OsGAP1)包含1个C2结构域,而含C2结构域的蛋白质是一类钙离子结合蛋白质,受钙信号的调控。本研究鉴定了水稻OsGAP1的由5个保守性天冬氨酸残基组成的阳离子结合区域。该区域可结合2个钙离子或者钾离子,且其结合钙离子的强度高于其结合钾离子的强度,但是不能结合镁离子。当将其中2个保守的天冬氨酸残基(Asp-23和Asp-28)突变为丙氨酸后,其对钙离子的结合能力减弱。对OsGAP1 C2结构域阳离子结合区域结合金属离子能力的研究,有助于加深对钙信号调控蛋白质的认识,为其在农业生产中的应用提供理论依据。展开更多
Alternative splicing can generate multiple mRNAs that differ in their untranslated regions or coding sequences,and these differences might affect mRNA stability or result in different protein isoforms with diverse fun...Alternative splicing can generate multiple mRNAs that differ in their untranslated regions or coding sequences,and these differences might affect mRNA stability or result in different protein isoforms with diverse functions and/or localizations.In this study,we isolated a sterile mutant in rice with abnormal meiosis of microspore mother cells and megaspore mother cells that carried a point mutation in OsRAD1 gene.Cloning of OsRAD1 cDNAs revealed three transcript variants,named as OsRAD1.1,OsRAD1.2 and OsRAD1.3,respectively,which were derived from alternative splicing of the last intron.Proteins derived from the three transcripts were mostly identical except the difference in the very C-terminal domain.The three transcripts exhibited similar expression patterns in various tissues,but the expression level of OsRAD1.1 was the highest.Specific knockout of OsRAD1.1 led to sterility,while knockout of OsRAD1.2 and OsRAD1.3 together did not change the plant fertility.Overexpression of OsRAD1.2 and OsRAD1.3 cDNAs in OsRAD1.1-specific mutant did not complement the plant fertility.Yeast two-hybrid assay showed that OsRAD1.1,but not OsRAD1.2 and OsRAD1.3,interacted with the three other meiosis proteins OsHUS1,OsRAD9 and OsRAD17,suggesting that the C-terminal domain of OsRAD1.1 is critical for the protein function.展开更多
Protein folding in the endoplasmic reticulum (ER) is a fundamental process in plant cells that is vulnerable to many environmental stresses. When unfolded or misfolded proteins accumulate in the ER, the well-conserv...Protein folding in the endoplasmic reticulum (ER) is a fundamental process in plant cells that is vulnerable to many environmental stresses. When unfolded or misfolded proteins accumulate in the ER, the well-conserved unfolded protein response (UPR) is initiated to mitigate the ER stress by enhancing the protein folding capability and/or accelerating the ER-associated protein degradation. Here, we report the conservation of the activation mechanism of OsbZIP74 (also known as OsbZIP50), an important ER stress regulator in monocot plant rice (Oryza sativa L.). Under normal conditions, OsbZIP74 mRNA encodes a basic leucine-zipper transcription factor with a putative transmembrane domain. When treating with ER stress-inducing agents such as tunicamycin and DTT, the conserved double stem-loop structures of OsbZIP74 mRNA are spliced out. Thereafter, the resulting new OsbZIP74 mRNA produces the nucleus-localized form of OsbZIP74 protein, eliminating the hydrophobic region. The activated form of OsbZIP74 has transcriptional activation activity in both yeast cells and Arabidopsis leaf protoplasts. The induction of OsbZIP74 splicing is much suppressed in the OsIRE1 knock- down rice plants, indicating the involvement of OslRE1 in OsbZIP74 splicing. We also demonstrate that the unconventional splicing of OsbZIP74 mRNA is associated with heat stress and salicylic acid, which is an important plant hormone in systemic acquired resistance against pathogen or parasite.展开更多
We cloned and characterized a rice gene OsBIABPI encoding an AMP-binding protein. The full-length cDNA of OsBIABP1 is 1912-bp long and is predicted to encode a 558-aa protein. OsBIABP1 contains a typical AMP-binding s...We cloned and characterized a rice gene OsBIABPI encoding an AMP-binding protein. The full-length cDNA of OsBIABP1 is 1912-bp long and is predicted to encode a 558-aa protein. OsBIABP1 contains a typical AMP-binding signature motif and shows high similarity to members of AMP-binding protein family. OsBIABP1 is expressed in stems, leaves and flowers of rice plants, but is not expressed, or expressed at a very low level, in rice roots. The expression of OsBIABP1 was induced by some defense-related signal molecules, e.g., salicylic acid (SA), benzothiadiazole, jasmonic acid (JA), and 1-amino cyclopropane- 1-carboxylic acid, which mediate SA- and JA/ethylene (ET)-dependent defense signaling pathways, respectively. Furthermore, the expression of OsBIABP1 is activated by the infection ofMagnaporthe oryzae, and the induced expression is quicker and stronger during early stages ofpathogenesis in incompatible interaction than that in compatible interaction between rice and M. oryzae. Our results suggest that OsBIABP1 may be a defense-related AMP-binding protein that is involved in the regulation of defense response through SA and/or JA/ET signaling pathways.展开更多
Many plant mltogen-actlvated protein klnases (MAPKs) play an important role In regulating responses to both ablotlc and biotic stresses. The first reported rice MAPK gene BWMK1 Is Induced by both rice blast (Magnap...Many plant mltogen-actlvated protein klnases (MAPKs) play an important role In regulating responses to both ablotlc and biotic stresses. The first reported rice MAPK gene BWMK1 Is Induced by both rice blast (Magnaporthe grisea) Infection and mechanical wounding. For further analysis of Its response to other environmental cues and plant hormones, such as jasmonlc acid (JA), salicylic acid (SA), and benzothladlazole (BTH), the promoter of BWMKf was fused with the coding region of the β-glucuronldase (GUS) reporter gene. Two promoter-GUS constructs with a 1.0- and 2.5-kb promoter fragment, respectively, were generated and transformed into the Japonica rice cultIvars TP309 and Zhonghua 11. Expression of GUS was Induced in the transgenic lines by cold, drought, dark, and JA. However, light, SA, and BTH treatments suppressed GUS expression. These results demonstrate that BWMK1 Is responsive to multiple ablotlc stresses and plant hormones and may play a role In cross-talk between different signaling pathways.展开更多
文摘Oryza sativa L. ssp. japonica and indica exhibit different sensitivity to photoinhibition and they show different stability of their core proteins D1 in the chloroplast photosystem Ⅱ. Using in situ hybridization, psbA, the gene encoding D1 protein of O. sativa ssp. japonica cv. 9516, and that of O. sativa ssp. indica cv. Shanyou 63 was cloned. As revealed by homology comparison of their sequences, the sequences are identical in the regions of promoter and 5′-UTR; differences are found in individual bases in the coding region all of which, being in the third position of respective codons, however, do not affect the amino acids coded finally; a difference is noted in the length of the oligo-U sequence in the region of 3′-UTR. It is thus apparent that, rather than a result of any difference in the amino acid sequences, the differences in the sensitivity to photoinhibition of D1 proteins between japonica and indica rice may be related to the upstream factors that regulate expression of psbA or to differences of photoprotective mechanisms.
基金supported by grants from Natural Science Foundation of Guangdong Province(Grant Nos.B030308008,2017A030310500 and A03013104)National Key Research and Development Plan Program(Grant Nos.2016YFD0101801 and 2016YFD0100406)+2 种基金Shenzhen Commission on Innovation and Technology Programs(Grant No.JCYJ20160229204920363)Guangzhou Science and Technology Innovation Commission(Grant No.201804010034)National Natural Science Foundation of China(Grant No.31500254).
文摘Alternative splicing can generate multiple mRNAs that differ in their untranslated regions or coding sequences,and these differences might affect mRNA stability or result in different protein isoforms with diverse functions and/or localizations.In this study,we isolated a sterile mutant in rice with abnormal meiosis of microspore mother cells and megaspore mother cells that carried a point mutation in OsRAD1 gene.Cloning of OsRAD1 cDNAs revealed three transcript variants,named as OsRAD1.1,OsRAD1.2 and OsRAD1.3,respectively,which were derived from alternative splicing of the last intron.Proteins derived from the three transcripts were mostly identical except the difference in the very C-terminal domain.The three transcripts exhibited similar expression patterns in various tissues,but the expression level of OsRAD1.1 was the highest.Specific knockout of OsRAD1.1 led to sterility,while knockout of OsRAD1.2 and OsRAD1.3 together did not change the plant fertility.Overexpression of OsRAD1.2 and OsRAD1.3 cDNAs in OsRAD1.1-specific mutant did not complement the plant fertility.Yeast two-hybrid assay showed that OsRAD1.1,but not OsRAD1.2 and OsRAD1.3,interacted with the three other meiosis proteins OsHUS1,OsRAD9 and OsRAD17,suggesting that the C-terminal domain of OsRAD1.1 is critical for the protein function.
基金This project is funded by the National Natural Science Foundation of China (31070233, 31171157), Shanghai Pujiang Talent Program (11PJ1400700), and partly supported by the National Basic Research Program of China (973 Program, 2012CB910500), all granted to J.X.L. ACKNOWLEDGMENTS We would also like thank Drs Yuhya Wakasa and Fumio Takaiwa for providing the OslREI transgenic rice seeds. No conflict of interest declared.
文摘Protein folding in the endoplasmic reticulum (ER) is a fundamental process in plant cells that is vulnerable to many environmental stresses. When unfolded or misfolded proteins accumulate in the ER, the well-conserved unfolded protein response (UPR) is initiated to mitigate the ER stress by enhancing the protein folding capability and/or accelerating the ER-associated protein degradation. Here, we report the conservation of the activation mechanism of OsbZIP74 (also known as OsbZIP50), an important ER stress regulator in monocot plant rice (Oryza sativa L.). Under normal conditions, OsbZIP74 mRNA encodes a basic leucine-zipper transcription factor with a putative transmembrane domain. When treating with ER stress-inducing agents such as tunicamycin and DTT, the conserved double stem-loop structures of OsbZIP74 mRNA are spliced out. Thereafter, the resulting new OsbZIP74 mRNA produces the nucleus-localized form of OsbZIP74 protein, eliminating the hydrophobic region. The activated form of OsbZIP74 has transcriptional activation activity in both yeast cells and Arabidopsis leaf protoplasts. The induction of OsbZIP74 splicing is much suppressed in the OsIRE1 knock- down rice plants, indicating the involvement of OslRE1 in OsbZIP74 splicing. We also demonstrate that the unconventional splicing of OsbZIP74 mRNA is associated with heat stress and salicylic acid, which is an important plant hormone in systemic acquired resistance against pathogen or parasite.
基金supported by the National Natural Science Foundation of China (No. 30771399)the High-Tech Research and Development Program (863) of China (No. 2007AA10Z140)the Research Fund for the Doctoral Program of Higher Education of the Ministry of Education of China (No. 20070335111)
文摘We cloned and characterized a rice gene OsBIABPI encoding an AMP-binding protein. The full-length cDNA of OsBIABP1 is 1912-bp long and is predicted to encode a 558-aa protein. OsBIABP1 contains a typical AMP-binding signature motif and shows high similarity to members of AMP-binding protein family. OsBIABP1 is expressed in stems, leaves and flowers of rice plants, but is not expressed, or expressed at a very low level, in rice roots. The expression of OsBIABP1 was induced by some defense-related signal molecules, e.g., salicylic acid (SA), benzothiadiazole, jasmonic acid (JA), and 1-amino cyclopropane- 1-carboxylic acid, which mediate SA- and JA/ethylene (ET)-dependent defense signaling pathways, respectively. Furthermore, the expression of OsBIABP1 is activated by the infection ofMagnaporthe oryzae, and the induced expression is quicker and stronger during early stages ofpathogenesis in incompatible interaction than that in compatible interaction between rice and M. oryzae. Our results suggest that OsBIABP1 may be a defense-related AMP-binding protein that is involved in the regulation of defense response through SA and/or JA/ET signaling pathways.
基金Supported, in part, by a Start-up Fund from the 0hio Agricultural Research and Development Center and the 0hio State University, the National Natural Science Foundation of China (30470990), and Scientific Research Fund of Hunan Provincial Education Department (04A024). Publication of this paper is supported by the National Natural Science Foundation of China (30624808).Acknowledgements The authors thank Drs Megan Griffith at Latrobe University in Australia and Beth Hazen at American Journal of Botany for suggestions and help with the preparation of the manuscript, and Maria Billizzi for technical assistance.
文摘Many plant mltogen-actlvated protein klnases (MAPKs) play an important role In regulating responses to both ablotlc and biotic stresses. The first reported rice MAPK gene BWMK1 Is Induced by both rice blast (Magnaporthe grisea) Infection and mechanical wounding. For further analysis of Its response to other environmental cues and plant hormones, such as jasmonlc acid (JA), salicylic acid (SA), and benzothladlazole (BTH), the promoter of BWMKf was fused with the coding region of the β-glucuronldase (GUS) reporter gene. Two promoter-GUS constructs with a 1.0- and 2.5-kb promoter fragment, respectively, were generated and transformed into the Japonica rice cultIvars TP309 and Zhonghua 11. Expression of GUS was Induced in the transgenic lines by cold, drought, dark, and JA. However, light, SA, and BTH treatments suppressed GUS expression. These results demonstrate that BWMK1 Is responsive to multiple ablotlc stresses and plant hormones and may play a role In cross-talk between different signaling pathways.