Songbiao Chen

Publications (13)66.15 Total impact

• Source

  Available from: Yuese Ning

  Article: The Magnaporthe oryzae Effector AvrPiz-t Targets the RING E3 Ubiquitin Ligase APIP6 to Suppress Pathogen-Associated Molecular Pattern-Triggered Immunity in Rice.

  Chan-Ho Park, Songbiao Chen, Gautam Shirsekar, Bo Zhou, Chang Hyun Khang, Pattavipha Songkumarn, Ahmed J Afzal, Yuese Ning, Ruyi Wang, Maria Bellizzi, Barbara Valent, Guo-Liang Wang
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  ABSTRACT: Although the functions of a few effector proteins produced by bacterial and oomycete plant pathogens have been elucidated in recent years, information for the vast majority of pathogen effectors is still lacking, particularly for those of plant-pathogenic fungi. Here, we show that the avirulence effector AvrPiz-t from the rice blast fungus Magnaporthe oryzae preferentially accumulates in the specialized structure called the biotrophic interfacial complex and is then translocated into rice (Oryza sativa) cells. Ectopic expression of AvrPiz-t in transgenic rice suppresses the flg22- and chitin-induced generation of reactive oxygen species (ROS) and enhances susceptibility to M. oryzae, indicating that AvrPiz-t functions to suppress pathogen-associated molecular pattern (PAMP)-triggered immunity in rice. Interaction assays show that AvrPiz-t suppresses the ubiquitin ligase activity of the rice RING E3 ubiquitin ligase APIP6 and that, in return, APIP6 ubiquitinates AvrPiz-t in vitro. Interestingly, agroinfection assays reveal that AvrPiz-t and AvrPiz-t Interacting Protein 6 (APIP6) are both degraded when coexpressed in Nicotiana benthamiana. Silencing of APIP6 in transgenic rice leads to a significant reduction of flg22-induced ROS generation, suppression of defense-related gene expression, and enhanced susceptibility of rice plants to M. oryzae. Taken together, our results reveal a mechanism in which a fungal effector targets the host ubiquitin proteasome system for the suppression of PAMP-triggered immunity in plants.
  The Plant Cell 11/2012; · 8.99 Impact Factor
• Article: Identification and Characterization of In-planta Expressed Secreted Effectors from Magnaporthe oryzae that Induce Cell Death in Rice.

  Songbiao Chen, Pattavipha Songkumarn, R Venu, Malali Gowda, Maria Bellizzi, Jinnan Hu, Wende Liu, D Ebbole, Tom Mitchell, Guo-Liang Wang
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  ABSTRACT: Interactions between rice and Magnaporthe oryzae involve the recognition of cellular components and the exchange of complex molecular signals from both partners. How these interactions occur in rice cells is still elusive. We employed robust-long serial analysis of gene expression (RL-SAGE), massively parallel signature sequencing (MPSS), and sequencing by synthesis (SBS), to examine transcriptome profiles of infected rice leaves. A total of 6,413 in-planta expressed fungal genes including 851 genes encoding predicted effector proteins were identified. We used a protoplast transient expression system to assess 42 of the predicted effector proteins for the ability to induce plant cell death. Ectopic expression assays identified five novel effectors that induced host cell death only when they contained the signal peptide for secretion to the extracellular space. The five apoplastic effectors induced cell death in a non-host-selective-manner. Although the five effectors are highly diverse in their sequences, the physiological basis of cell death induced by each was similar. This study demonstrates that our integrative genomic approach is effective for the identification of in-planta expressed cell death-inducing effectors from M. oryzae that may play an important role facilitating colonization and fungal growth during infection.
  Molecular Plant-Microbe Interactions 10/2012; · 4.43 Impact Factor
• Source

  Available from: Gyan Mishra

  Article: A 5-methylcytosine DNA glycosylase/lyase demethylates the retrotransposon Tos17 and promotes its transposition in rice.

  Honggui La, Bo Ding, Gyan P Mishra, Bo Zhou, Hongmei Yang, Maria del Rosario Bellizzi, Songbiao Chen, Blake C Meyers, Zhaohua Peng, Jian-Kang Zhu, Guo-Liang Wang
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  ABSTRACT: DNA 5-methylcytosine (5-meC) is an important epigenetic mark for transcriptional gene silencing in many eukaryotes. In Arabidopsis, 5-meC DNA glycosylase/lyases actively remove 5-meC to counteract transcriptional gene silencing in a locus-specific manner, and have been suggested to maintain the expression of transposons. However, it is unclear whether plant DNA demethylases can promote the transposition of transposons. Here we report the functional characterization of the DNA glycosylase/lyase DNG701 in rice. DNG701 encodes a large (1,812 amino acid residues) DNA glycosylase domain protein. Recombinant DNG701 protein showed 5-meC DNA glycosylase and lyase activities in vitro. Knockout or knockdown of DNG701 in rice plants led to DNA hypermethylation and reduced expression of the retrotransposon Tos17. Tos17 showed less transposition in calli derived from dng701 knockout mutant seeds compared with that in wild-type calli. Overexpression of DNG701 in both rice calli and transgenic plants substantially reduced DNA methylation levels of Tos17 and enhanced its expression. The overexpression also led to more frequent transposition of Tos17 in calli. Our results demonstrate that rice DNG701 is a 5-meC DNA glycosylase/lyase responsible for the demethylation of Tos17 and this DNA demethylase plays a critical role in promoting Tos17 transposition in rice calli.
  Proceedings of the National Academy of Sciences 09/2011; 108(37):15498-503. · 9.68 Impact Factor
• Source

  Available from: Yuese Ning

  Article: The SINA E3 ligase OsDIS1 negatively regulates drought response in rice.

  Yuese Ning, Chachawan Jantasuriyarat, Qingzhen Zhao, Huawei Zhang, Songbiao Chen, Jinling Liu, Lijing Liu, Sanyuan Tang, Chan Ho Park, Xuejun Wang, Xionglun Liu, Liangying Dai, Qi Xie, Guo-Liang Wang
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  ABSTRACT: Ubiquitin-regulated protein degradation is a critical regulatory mechanism that controls a wide range of biological processes in plants. Here, we report that OsDIS1 (for Oryza sativa drought-induced SINA protein 1), a C3HC4 RING finger E3 ligase, is involved in drought-stress signal transduction in rice (O. sativa). The expression of OsDIS1 was up-regulated by drought treatment. In vitro ubiquitination assays showed that OsDIS1 possessed E3 ubiquitin ligase activity and that the conserved region of the RING finger was required for the activity. Transient expression assays in Nicotiana benthamiana leaves and rice protoplasts indicated that OsDIS1 was localized predominantly in the nucleus. Overexpression of OsDIS1 reduced drought tolerance in transgenic rice plants, while RNA interference silencing of OsDIS1 enhanced drought tolerance. Microarray analysis revealed that a large number of drought-responsive genes were induced or suppressed in the OsDIS1 overexpression plants under normal and drought conditions. Yeast two-hybrid screening showed that OsDIS1 interacted with OsNek6 (for O. sativa NIMA-related kinase 6), a tubulin complex-related serine/threonine protein kinase. Coexpression assays in N. benthamiana leaves indicated that OsNek6 was degraded by OsDIS1 via the 26S proteasome-dependent pathway and that this degradation was abolished by the OsDIS1(H71Y) mutation, which is essential for its E3 ligase activity. Together, these results demonstrate that OsDIS1 plays a negative role in drought stress tolerance through transcriptional regulation of diverse stress-related genes and possibly through posttranslational regulation of OsNek6 in rice.
  Plant physiology 06/2011; 157(1):242-55. · 6.53 Impact Factor
• Article: A versatile zero background T-vector system for gene cloning and functional genomics.

  Songbiao Chen, Pattavipha Songkumarn, Jianli Liu, Guo-Liang Wang
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  ABSTRACT: With the recent availability of complete genomic sequences of many organisms, high-throughput and cost-efficient systems for gene cloning and functional analysis are in great demand. Although site-specific recombination-based cloning systems, such as Gateway cloning technology, are extremely useful for efficient transfer of DNA fragments into multiple destination vectors, the two-step cloning process is time consuming and expensive. Here, we report a zero background TA cloning system that provides simple and high-efficiency direct cloning of PCR-amplified DNA fragments with almost no self-ligation. The improved T-vector system takes advantage of the restriction enzyme XcmI to generate a T-overhang after digestion and the negative selection marker gene ccdB to eliminate the self-ligation background after transformation. We demonstrate the feasibility and flexibility of the technology by developing a set of transient and stable transformation vectors for constitutive gene expression, gene silencing, protein tagging, protein subcellular localization detection, and promoter fragment activity analysis in plants. Because the system can be easily adapted for developing specialized expression vectors for other organisms, zero background TA provides a general, cost-efficient, and high-throughput platform that complements the Gateway cloning system for gene cloning and functional genomics.
  Plant physiology 05/2009; 150(3):1111-21. · 6.53 Impact Factor
• Chapter: Isolation and Functional Analysis of Putative Effectors from Magnaporthe oryzae Using Integrated Genomic Approaches

  Songbiao Chen, Malali Gowda, R.C. Venu, Pattavipha Songkumarn, Chan Ho Park, Maria Bellizzi, Daniel J. Ebbole, Guo-Liang Wang
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  ABSTRACT: Rice blast disease, caused by the fungus Magnaporthe oryzae, is a leading constraint to rice production and is a serious threat to food security worldwide. To elucidate the function of effector proteins from M. oryzae in pathogenesis and interaction with the host, we have performed RL-SAGE and MPSS approaches to study the gene expression profiles of M. oryzae during the interaction. The RL-SAGE and MPSS analyses identified 3,441 and 3,004 annotated M. oryzae genes from blast infected rice leaf tissues, respectively. Among them, 217 genes encoding putative secreted proteins, which may play important roles as effectors, were identified. We developed a highly efficient transient protoplast system for gene functional analysis in rice. We present here the detailed procedure of the rice protoplast transient expression system and show the examples of using the system for functional analysis of putative effectors from M. oryzae. The combination of RL-SAGE/MPSS genomic profiling approaches with a protoplast functional assay system has provided an efficient approach for large-scale isolation and analysis of effectors from M. oryzae.
  12/2008: pages 93-103;
• Source

  Available from: Miguel E. Vega-Sanchez

  Article: SPIN1, a K homology domain protein negatively regulated and ubiquitinated by the E3 ubiquitin ligase SPL11, is involved in flowering time control in rice.

  Miguel E Vega-Sánchez, Lirong Zeng, Songbiao Chen, Hei Leung, Guo-Liang Wang
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  ABSTRACT: The rice (Oryza sativa) E3 ligase SPOTTED LEAF11 (SPL11) negatively regulates programmed cell death and disease resistance. We demonstrate here that SPL11 also regulates flowering via interaction with SPIN1 (for SPL11-interacting protein1), a Signal Transduction and Activation of RNA family member. SPIN1 binds RNA and DNA in vitro and interacts with SPL11 in the nucleus. Spl11 mutants have delayed flowering under long-day conditions. Spin1 overexpression causes late flowering independently of daylength; expression analyses of flowering marker genes in these lines suggested that SPIN1 represses flowering by downregulating the flowering promoter gene Heading date3a (Hd3a) via Hd1-dependent mechanisms in short days and by targeting Hd1-independent factors in long days. Both Spin1 and Spl11 are regulated diurnally in opposing phases. SPL11 negatively regulates Spin1 transcript levels, while SPIN1 also affects Spl11 expression. Moreover, we show that coincidence of high accumulation of Spin1 mRNA with the light in the morning and early evening is needed to repress flowering. SPIN1 is monoubiquitinated by SPL11, suggesting that it is not targeted for degradation. Our data are consistent with a model in which SPIN1 acts as a negative regulator of flowering that itself is negatively regulated by SPL11, possibly via ubiquitination.
  The Plant Cell 07/2008; 20(6):1456-69. · 8.99 Impact Factor
• Article: Magnaporthe grisea infection triggers RNA variation and antisense transcript expression in rice.

  Malali Gowda, R-C Venu, Huameng Li, Chatchawan Jantasuriyarat, Songbiao Chen, Maria Bellizzi, Vishal Pampanwar, HyeRan Kim, Ralph A Dean, Eric Stahlberg, Rod Wing, Cari Soderlund, Guo-Liang Wang
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  ABSTRACT: Rice blast disease, caused by the fungal pathogen Magnaporthe grisea, is an excellent model system to study plant-fungal interactions and host defense responses. In this study, comprehensive analysis of the rice (Oryza sativa) transcriptome after M. grisea infection was conducted using robust-long serial analysis of gene expression. A total of 83,382 distinct 21-bp robust-long serial analysis of gene expression tags were identified from 627,262 individual tags isolated from the resistant (R), susceptible (S), and control (C) libraries. Sequence analysis revealed that the tags in the R and S libraries had a significant reduced matching rate to the rice genomic and expressed sequences in comparison to the C library. The high level of one-nucleotide mismatches of the R and S library tags was due to nucleotide conversions. The A-to-G and U-to-C nucleotide conversions were the most predominant types, which were induced in the M. grisea-infected plants. Reverse transcription-polymerase chain reaction analysis showed that expression of the adenine deaminase and cytidine deaminase genes was highly induced after inoculation. In addition, many antisense transcripts were induced in infected plants and expression of four antisense transcripts was confirmed by strand-specific reverse transcription-polymerase chain reaction. These results demonstrate that there is a series of dynamic and complex transcript modifications and changes in the rice transcriptome at the M. grisea early infection stages.
  Plant physiology 06/2007; 144(1):524-33. · 6.53 Impact Factor
• Article: A highly efficient transient protoplast system for analyzing defence gene expression and protein-protein interactions in rice.

  Songbiao Chen, Lizen Tao, Lirong Zeng, Miguel E Vega-Sanchez, Kenji Umemura, Guo-Liang Wang
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  ABSTRACT: SUMMARY The transient assay system based on mesophyll or cultured cell-derived protoplasts has been exploited in several plant species and has become a powerful tool for rapid gene functional analysis and biochemical manipulations. However, the system has not been widely used in rice owing to the difficulties in large-scale isolation of viable rice protoplasts from leaves or suspension-cultured cells. Here, we describe a significantly improved method to isolate a large number of protoplasts from stem and sheath tissues of both young and mature plants. High-level coexpression of multiple constructs and efficient suppression of exogenous and endogenous genes were observed in the stem- and sheath-derived protoplasts. A transient green fluorescent protein and luciferase-based reporter system for defence-related genes expression analysis has been established, which is useful for screening and characterizing genes involved in rice defence signalling pathways. Furthermore, a protoplast-based bimolecular fluorescence complementation (BiFC) system for the detection of protein-protein interactions in living rice cells was developed. The YFP complementation of two split-YFP halves mediated by homodimerization of the GUS and SPIN1, a cell-death related protein, was observed in transfected protoplasts. In combination with genetic, genomic and proteomic approaches, the established versatile protoplast transient assay system will facilitate large-scale functional analysis of defence-related genes in rice.
  Molecular Plant Pathology 09/2006; 7(5):417-27. · 3.90 Impact Factor
• Article: Cloning and Functional Analysis of the Bifunctional Agglutinin/Trypsin Inhibitor from Helianthus tuberosus L.

  Tuanjie Chang, Hongli Zhai, Songbiao Chen, Guisheng Song, Honglin Xu, Xiaoli Wei, Zhen Zhu
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  ABSTRACT: In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively (GenBank accession numbers AF477031-AF477034), were isolated from a tuber cDNA expression library of Helianthus tuberosus L. Sequence analysis revealed that all four cDNAs contain an open reading frame of 444 bp, coding a polypeptide of 147 amino acid residues, and that the sequences of the cDNAs are very similar to those of the mannose-binding agglutinin genes of the jacalin-related family. In hemagglutination reactions and hapten inhibition assays, affinity-purified HTA (Helianthus tuberosus agglutinin) from induced Escherichia coli BL21(DE3) expressing GST-HTA shows hemagglutination ability and a higher carbohydrate-binding ability for mannose than other tested sugars. Trypsin inhibitory activity was detected in the crude extracts of induced E. coli BL21(DE3) expressing HTA, and was further verified by trypsin inhibitory activity staining on native polyacrylamide gel. The mechanism of interaction between HTA and trypsin was studied by molecular modeling. We found that plenty of hydrogen bonds and electrostatic interactions can be formed between the supposed binding sites of HTA-b and the active site of trypsin, and that a stable HTA/trypsin complex can be formed. The results above imply that HTA might be a bifunctional protein with carbohydrate-binding activity and trypsin inhibitory activity. Moreover, Northern blotting analysis demonstrated that hta is predominantly expressed in tubers of H. tuberosus, very weakly expressed in stems, but not expressed at all in other tissues. Southern blotting analysis indicated that hta is encoded by a multi-gene family. The insect resistance traits have been described in another paper.(Managing editor: Li-Hui Zhao)
  Journal of Integrative Plant Biology 08/2006; 48(8):971 - 982. · 2.53 Impact Factor
• Article: Green fluorescent protein as a vital elimination marker to easily screen marker-free transgenic progeny derived from plants co-transformed with a double T-DNA binary vector system.

  Songbiao Chen, Xugang Li, Xiang Liu, Hongling Xu, Kun Meng, Guifang Xiao, Xiaoli Wei, Feng Wang, Zhen Zhu
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  ABSTRACT: We investigated the potential of a novel double T-DNA vector for generating marker-free transgenic plants. Co-transformation methods using a double T-DNA vector or using mixture of two Agrobacterium tumefaciens strains were compared, and showed that the double T-DNA vector method could produce marker-free transgenic tobacco (Nicotiana tabacum L.) plants more efficiently. A dual marker double T-DNA vector was then constructed by assembling the green fluorescent protein (GFP) gene mgfp5 and the neomycin phosphotransferase gene nptII into the same T-DNA. The frequency of co-transformants produced by this vector was 56.3%. Co-expression of mgfp5 and nptII was found in 28 out of 29 T1 lines, and segregation of the reporter beta-glucuronidase gene, gusA, from mgfp5 to nptII was found in 12 out of 29 T1 lines. Therefore, GFP could be used as a vital marker to improve the transformation efficiency and to easily monitor the segregation of marker genes, thus facilitating screening of marker-free progeny.
  Plant Cell Reports 03/2005; 23(9):625-31. · 2.27 Impact Factor
• Article: A metallothionein-like gene htMT2 strongly expressed in internodes and nodes of Helianthus tuberosus and effects of metal ion treatment on its expression.

  Tuanjie Chang, Xiang Liu, Honglin Xu, Kun Meng, Songbiao Chen, Zhen Zhu
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  ABSTRACT: A cDNA sequence encoding a type-2 metallothionein (MT)-like protein, designated htMT2, was isolated from a Helianthus tuberosus L. tuber cDNA library. The isolated cDNA is 509 bp, coding a 7.8-kDa polypeptide. Two partial genomic fragments covering the open reading frame of htMT2 were cloned by PCR. The fragments htMTG-1 (986 bp) and htMTG-2 (982 bp) contain three exons and two introns. The N- and C-terminal domains of the predicted polypeptide have eight and seven cysteine residues, separated by a central cysteine-free spacer. Sequence alignment revealed that the predicted protein was homologous to type-2 MTs of plants. Southern blot analysis indicated that htMT2 is encoded by a small multi-gene family in the H. tuberosus genome. Northern blot analysis showed that htMT2 transcripts were predominantly expressed in internodes and nodes, but were low in leaves, leafstalks, tubers and young roots, and none was detected in roots. Treatment with Cu(2+) reduced the expression of htMT2 in internodes, nodes, leaves and leafstalks. In addition, the expression levels in internodes and nodes share an inverse relationship with the concentrations of Cu(2+). In internodes and nodes, treatment with Zn(2+) at 10 and 100 microM reduced the expression levels of htMT2, and 1000 microM Zn(2+)reduced it to the lowest level, but 500 microM Zn(2+) had little effect. The expressions of htMT2 in different tissues were not appreciably affected by heat shock. It is suggested that HtMT2 might be involved in the transport or availability of Cu(2+) and Zn(2+) to some apo-metal enzymes or apo-metal proteins.
  Planta 02/2004; 218(3):449-55. · 3.00 Impact Factor
• Article: Transformation of tobacco with genes encoding Helianthus tuberosus agglutinin (HTA) confers resistance to peach-potato aphid (Myzus persicae).

  Tuanjie Chang, Lei Chen, Songbiao Chen, Huaya Cai, Xiang Liu, Guifang Xiao, Zhen Zhu
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  ABSTRACT: The effects of the hta gene encoding Helianthus tuberosus agglutinin (HTA) on an insect in the order Homoptera were investigated. Homologous cDNAs of hta-a, hta-b, hta-c and hta-d with CaMV35S as promoter were introduced into tobacco via Agrobacterium tumefaciens. Southern blot results showed that the exogenous hta gene was inserted into the genome of host plants, and northern blot analysis confirmed that hta was expressed in transgenic plants. A bioassay with peach-potato aphid (Myzus persicae) demonstrated that transgenic plants had deleterious effects on the insect. The average population of aphids fed on transgenic T0 plants during an 11-day assay decreased by 70%, compared controls. In transgenic plants of T1 generation, aphid fecundity inhibitions were 53.0% (hta-b) and 64.6% (hta-c), respectively. The development of aphids was notably retarded. We conclude that hta could be a novel and promising candidate for plant transgenic engineering against homopteran insect pests.
  Transgenic Research 11/2003; 12(5):607-14. · 2.75 Impact Factor