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液泡加工酶VPE正向调节植物对寄生疫霉的抗病性及细胞死亡

GAO Xian-xian, TANG Ya-ling, SHI Qing-yao, WEI Yu-shu, WANG Xiao-xue, SHAN Wei-xing, QIANG Xiao-yu

Journal of Integrative Agriculture 2023, 22 (5): 1424-1433. DOI: 10.1016/j.jia.2022.08.124

摘要（211）

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疫霉属卵菌引致马铃薯晚疫病等作物灾难性病害，严重威胁作物可持续生产。由于其独特的遗传变异机制，导致作物品种抗病性丧失问题极为突出，因此亟需挖掘和探索新型抗病基因资源及其免疫调控机理，并将之有效应用于作物抗病分子育种。拟南芥RTP1 (Resistance to Phytophthora parasitica 1)是前期研究鉴定获得的免疫负调控因子，RTP1基因缺失的拟南芥rtp1突变体植株呈现出对多种病原菌的抗病性，并在病菌侵染初期产生快速细胞死亡和活性氧累积。基于细胞死亡在植物免疫中的重要作用，本研究旨在探究RTP1介导的植物细胞死亡机理，鉴定到一类液泡加工酶γVPE能够影响rtp1突变体响应寄生疫霉侵染而引发的抗病反应及细胞死亡。以寄生疫霉侵染的拟南芥野生型Columbia-0 (Col-0)和rtp1突变体植株为材料，通过实时定量PCR分析VPE基因表达模式，并利用VPE/Caspase-1蛋白酶的特异荧光底物分析酶活性水平，结果表明，相较于野生型Col-0，寄生疫霉侵染的拟南芥rtp1突变体中γVPE基因上调表达，并伴随着升高的VPE/caspase-1酶活性水平。进一步利用特异酶活性抑制剂，结果揭示了拟南芥植株响应寄生疫霉侵染而产生的细胞死亡以及rtp1突变体对寄生疫霉的抗病性均依赖于VPE/caspase-1酶活性。利用拟南芥γvpe突变体植株或农杆菌介导的AtγVPE瞬时过表达烟草叶片进行接菌表型分析，结果证明了AtγVPE能够正向调节植物对寄生疫霉的抗病性。综上所述，本研究不仅揭示了γVPE是植物响应寄生疫霉侵染过程中参与调节植物抗病反应和细胞死亡的关键因子，还有助于深入理解感病因子RTP1介导的细胞死亡调节机制，为作物抗病分子育种提供了新型基因资源与新思路。

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2. Transcriptomic analyses reveal new genes and networks response to H5N1 influenza viruses in duck (Anas platyrhynchos)

HUANG Yin-hua, FENG Hua-peng, HUANG Li-ren, YI Kang, RONG En-guang, CHEN Xiao-yun, LI Jian-wen, WANG Zeng, ZHU Peng-yang, LIU Xiao-juan, WANG Xiao-xue, HU Jia-xiang, LIU Xin, CHEN Hua-lan, WANG Jun...

Journal of Integrative Agriculture 2019, 18 (7): 1460-1472. DOI: 10.1016/S2095-3119(19)62646-8

摘要（205）

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H5N1 influenza represents one of the great challenges to public health. Some H5N1 viruses (i.e., A/goose/Hubei/65/05, GS/65) are weakly pathogenic, while the others (i.e., A/duck/Hubei/49/05, DK/49) are highly pathogenic to their natural hosts. Here, we performed brain and spleen transcriptomic analyses of control ducks and ones infected by the DK/49 or the GS/65 H5N1 virus. We demonstrated that, compared to the GS/65 virus, the DK/49 virus infection changed more numerous immune genes’ expression and caused continuous increasing of immune pathways (i.e., RIG-I and MDA5) in ducks. We found that both H5N1 virus strains might escape or subvert host immune response through affecting alternative translation of immune genes, while the DK/49 virus seemed to induce alternative translation of more immune genes than the GS/65 virus. We also identified five co-expressional modules associated with H5N1 virus replication through the weight correlation network analysis (WGCNA). Moreover, we first demonstrated that the duck BCL2L15 and DCSTAMP in one of these five modules inhibited both the highly pathogenic and weakly pathogenic H5N1 virus replication efficiently. These analyses, in combination with our comprehensive transcriptomic data, provided global view of the molecular architecture for the interaction between host and H5N1 viruses.

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3. Identification of novel genes associated with duck OASL in response to influenza A virus

WANG Xiao-xue, LU Chang, RONG En-guang, HU Jia-xiang, XING Yan-ling, LIU Zheng-yu, GAO Chu-ze, LIU Jin-hua, HUANG Yin-hua

Journal of Integrative Agriculture 2019, 18 (7): 1451-1459. DOI: 10.1016/S2095-3119(19)62685-7

摘要（201）

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2´-5´-Oligoadenylate synthetase like protein (OASL) plays a key role in response to viral infections through selectively activating the OAS/RNase L or OASL/RIG-I signaling pathway. Although classic pathway of OASL is well-known, its regulated genes or co-actors are largely unknown. To study the possible molecular mechanism of duck OASL (dOASL), we performed RNA-sequencing (RNA-seq) and immunoprecipitation and mass spectrometry (IP-MS) at the level of mRNA and protein, respectively. For RNA-seq, we used DF1 cell lines (DF1^dOASL+/+,DF1^{cOASL–/–,}and DF1) with or without the CK/0513 H5N1 virus (A/chicken/huabei/0513/2007) infection. 1 737 differentially expressed genes (DEGs) were identified as candidate target genes regulated by dOASL. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Weighted Correlation Network Analysis (WGCNA) were performed. We identified one important yellow co-expression module correlated with antiviral immune response. In this module, Ankyrin repeat and FYVE domain containing 1 (ANKFY1), harboring a BTB domain similar to the methyl CpG-binding protein 1 (MBD1) which bound to OASL in human, was regulated by dOASL. At protein level, 133 host proteins were detected. Interestingly, ANKFY1 was one of them binding to dOASL protein. Further phylogenomic and chromosomal syntenic analysis demonstrated MBD1 was absent in birds, while mammals retained. It is suggested that OASL-ANKFY1 interaction might act as a compensatory mechanism to regulate gene expression in birds. Our findings will provide a useful resource for the molecular mechanism research of dOASL.

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4. Genetic diversity and elite gene introgression reveal the japonica rice breeding in northern China

LIU Dan, WANG Jia-yu, WANG Xiao-xue, YANG Xian-li, SUN Jian, CHEN Wen-fu

Journal of Integrative Agriculture 2015, 14 (5): 811-822. DOI: 10.1016/S2095-3119(14)60898-4

摘要（2350）

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Abundant genetic diversity and rational population structure of germplasm benefit crop breeding greatly. To investigate genetic variation among geographically diverse set of japonica germplasm, we analyzed 233 japonica rice cultivars collected from Liaoning, Jilin and Heilongjiang provinces of China, which were released from 1970 to 2011 by using 62 simple sequence repeat (SSR) markers and 8 functional gene tags related to yield. A total of 195 alleles (Na) were detected with an average of 3.61 per locus, indicating a low level of genetic diversity level among all individuals. The genetic diversity of the cultivars from Jilin Province was the highest among the three geographic distribution zones. Moreover, the genetic diversity was increased slightly with the released period of cultivars from 1970 to 2011. The analysis of molecular variance (AMOVA) revealed that genetic differentiation was more diverse within the populations than that among the populations. The neighbor-joining (NJ) tree indicated that cultivar clusters based on geographic distribution represented three independent groups, among which the cluster of cultivars from Heilongjiang is distinctly different to the cluster of cultivars from Liaoning. For the examined functional genes, two or three allelic variations for each were detected, except for IPA1 and GW2, and most of elite genes had been introgressed in modern japonica rice varieties. These results provide a valuable evaluation for genetic backgrounds of current japonica rice and will be used directly for japonica rice breeding in future.

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