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Plant-LncPipe: a computational pipeline providing significant improvement in plant lncRNA identification. Plant-LncPipe:一种可显著改进植物 lncRNA 鉴定的计算管道。
Pub Date : 2024-02-08 eCollection Date: 2024-04-01 DOI: 10.1093/hr/uhae041
Xue-Chan Tian, Zhao-Yang Chen, Shuai Nie, Tian-Le Shi, Xue-Mei Yan, Yu-Tao Bao, Zhi-Chao Li, Hai-Yao Ma, Kai-Hua Jia, Wei Zhao, Jian-Feng Mao

Long non-coding RNAs (lncRNAs) play essential roles in various biological processes, such as chromatin remodeling, post-transcriptional regulation, and epigenetic modifications. Despite their critical functions in regulating plant growth, root development, and seed dormancy, the identification of plant lncRNAs remains a challenge due to the scarcity of specific and extensively tested identification methods. Most mainstream machine learning-based methods used for plant lncRNA identification were initially developed using human or other animal datasets, and their accuracy and effectiveness in predicting plant lncRNAs have not been fully evaluated or exploited. To overcome this limitation, we retrained several models, including CPAT, PLEK, and LncFinder, using plant datasets and compared their performance with mainstream lncRNA prediction tools such as CPC2, CNCI, RNAplonc, and LncADeep. Retraining these models significantly improved their performance, and two of the retrained models, LncFinder-plant and CPAT-plant, alongside their ensemble, emerged as the most suitable tools for plant lncRNA identification. This underscores the importance of model retraining in tackling the challenges associated with plant lncRNA identification. Finally, we developed a pipeline (Plant-LncPipe) that incorporates an ensemble of the two best-performing models and covers the entire data analysis process, including reads mapping, transcript assembly, lncRNA identification, classification, and origin, for the efficient identification of lncRNAs in plants. The pipeline, Plant-LncPipe, is available at: https://github.com/xuechantian/Plant-LncRNA-pipline.

长非编码 RNA(lncRNA)在染色质重塑、转录后调控和表观遗传修饰等多种生物过程中发挥着重要作用。尽管长编码 RNA 在调控植物生长、根系发育和种子休眠等方面具有重要功能,但由于缺乏特异性的、经过广泛测试的鉴定方法,植物长编码 RNA 的鉴定仍然是一项挑战。大多数用于植物lncRNA鉴定的基于机器学习的主流方法最初都是利用人类或其他动物数据集开发的,它们在预测植物lncRNA方面的准确性和有效性尚未得到充分评估或利用。为了克服这一局限性,我们使用植物数据集重新训练了 CPAT、PLEK 和 LncFinder 等几个模型,并将它们的性能与 CPC2、CNCI、RNAplonc 和 LncADeep 等主流 lncRNA 预测工具进行了比较。对这些模型的再训练大大提高了它们的性能,其中两个再训练模型--LncFinder-plant 和 CPAT-plant 以及它们的集合成为最适合植物 lncRNA 鉴定的工具。这凸显了模型再训练在应对植物 lncRNA 鉴定相关挑战中的重要性。最后,我们开发了一个管道(Plant-LncPipe),它包含了两个表现最好的模型的集合,涵盖了整个数据分析过程,包括读数映射、转录本组装、lncRNA 鉴定、分类和起源,以高效鉴定植物中的 lncRNA。该管道Plant-LncPipe可在以下网址获取:https://github.com/xuechantian/Plant-LncRNA-pipline。
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引用次数: 0
Dissecting the complex genetic basis of pre- and post-harvest traits in Vitis vinifera L. using genome-wide association studies. 利用全基因组关联研究剖析葡萄采收前后性状的复杂遗传基础。
Pub Date : 2024-01-03 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad283
Julian García-Abadillo, Paola Barba, Tiago Carvalho, Viviana Sosa-Zuñiga, Roberto Lozano, Humberto Fanelli Carvalho, Miguel Garcia-Rojas, Erika Salazar, Julio Isidro Y Sánchez

Addressing the pressing challenges in agriculture necessitates swift advancements in breeding programs, particularly for perennial crops like grapevines. Moving beyond the traditional biparental quantitative trait loci (QTL) mapping, we conducted a genome-wide association study (GWAS) encompassing 588 Vitis vinifera L. cultivars from a Chilean breeding program, spanning three seasons and testing 13 key yield-related traits. A strong candidate gene, Vitvi11g000454, located on chromosome 11 and related to plant response to biotic and abiotic stresses through jasmonic acid signaling, was associated with berry width and holds potential for enhancing berry size in grape breeding. We also mapped novel QTL associated with post-harvest traits across chromosomes 2, 4, 9, 11, 15, 18, and 19, broadening our grasp on the genetic intricacies dictating fruit post-harvest behavior, including decay, shriveling, and weight loss. Leveraging gene ontology annotations, we drew parallels between traits and scrutinized candidate genes, laying a robust groundwork for future trait-feature identification endeavors in plant breeding. We also highlighted the importance of carefully considering the choice of the response variable in GWAS analyses, as the use of best linear unbiased estimators (BLUEs) corrections in our study may have led to the suppression of some common QTL in grapevine traits. Our results underscore the imperative of pioneering non-destructive evaluation techniques for long-term conservation traits, offering grape breeders and cultivators insights to improve post-harvest table grape quality and minimize waste.

要应对农业领域的紧迫挑战,就必须迅速推进育种计划,尤其是像葡萄这样的多年生作物。我们超越了传统的双亲数量性状位点(QTL)图谱,开展了一项全基因组关联研究(GWAS),涵盖了智利育种计划中的 588 个葡萄品种,跨越三个季节,测试了 13 个与产量相关的关键性状。一个强候选基因 Vitvi11g000454 位于第 11 号染色体上,通过茉莉酸信号转导与植物对生物和非生物胁迫的响应有关,该基因与浆果宽度相关,具有在葡萄育种中提高浆果大小的潜力。我们还在 2、4、9、11、15、18 和 19 号染色体上绘制了与采后性状相关的新型 QTL,从而拓宽了我们对决定果实采后行为(包括腐烂、干瘪和重量损失)的遗传复杂性的掌握。利用基因本体注释,我们得出了性状与候选基因之间的相似之处,为今后植物育种中的性状特征鉴定工作奠定了坚实的基础。我们还强调了在 GWAS 分析中仔细考虑响应变量选择的重要性,因为在我们的研究中使用最佳线性无偏估计值(BLUEs)校正可能会导致葡萄性状中一些常见 QTL 的抑制。我们的研究结果强调了针对长期保存性状开创非破坏性评价技术的必要性,为葡萄育种者和栽培者提供了提高采后鲜食葡萄质量和最大限度减少浪费的真知灼见。
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引用次数: 0
Exploring large-scale gene coexpression networks in peach (Prunus persica L.): a new tool for predicting gene function. 探索桃(Prunus persica L.)中的大规模基因共表达网络:预测基因功能的新工具。
Pub Date : 2024-01-02 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad294
Felipe Pérez de Los Cobos, Beatriz E García-Gómez, Luis Orduña-Rubio, Ignasi Batlle, Pere Arús, José Tomás Matus, Iban Eduardo

Peach is a model for Prunus genetics and genomics, however, identifying and validating genes associated to peach breeding traits is a complex task. A gene coexpression network (GCN) capable of capturing stable gene-gene relationships would help researchers overcome the intrinsic limitations of peach genetics and genomics approaches and outline future research opportunities. In this study, we created four GCNs from 604 Illumina RNA-Seq libraries. We evaluated the performance of every GCN in predicting functional annotations using an algorithm based on the 'guilty-by-association' principle. The GCN with the best performance was COO300, encompassing 21 956 genes. To validate its performance predicting gene function, we performed two case studies. In case study 1, we used two genes involved in fruit flesh softening: the endopolygalacturonases PpPG21 and PpPG22. Genes coexpressing with both genes were extracted and referred to as melting flesh (MF) network. Finally, we performed an enrichment analysis of MF network and compared the results with the current knowledge regarding peach fruit softening. The MF network mostly included genes involved in cell wall expansion and remodeling, and with expressions triggered by ripening-related phytohormones, such as ethylene, auxin, and methyl jasmonate. In case study 2, we explored potential targets of the anthocyanin regulator PpMYB10.1 by comparing its gene-centered coexpression network with that of its grapevine orthologues, identifying a common regulatory network. These results validated COO300 as a powerful tool for peach and Prunus research. This network, renamed as PeachGCN v1.0, and the scripts required to perform a function prediction analysis are available at https://github.com/felipecobos/PeachGCN.

水蜜桃是李属植物遗传学和基因组学的典范,然而,识别和验证与水蜜桃育种性状相关的基因是一项复杂的任务。能够捕捉稳定的基因-基因关系的基因共表达网络(GCN)将有助于研究人员克服桃遗传学和基因组学方法的内在局限性,并勾勒出未来的研究机会。在本研究中,我们从 604 个 Illumina RNA-Seq 文库中创建了四个 GCN。我们使用基于 "有罪关联 "原则的算法评估了每个 GCN 在预测功能注释方面的性能。性能最好的 GCN 是 COO300,包含 21 956 个基因。为了验证其预测基因功能的性能,我们进行了两项案例研究。在案例研究 1 中,我们使用了两个参与果肉软化的基因:内多聚半乳糖醛酸酶 PpPG21 和 PpPG22。我们提取了与这两个基因共表达的基因,并将其称为果肉融化(MF)网络。最后,我们对 MF 网络进行了富集分析,并将分析结果与目前有关桃果软化的知识进行了比较。MF 网络主要包括参与细胞壁扩张和重塑的基因,以及由成熟相关植物激素(如乙烯、辅助素和茉莉酸甲酯)引发表达的基因。在案例研究 2 中,我们通过比较花青素调控因子 PpMYB10.1 与葡萄同源物的基因中心共表达网络,发现了一个共同的调控网络,从而探索了花青素调控因子 PpMYB10.1 的潜在靶标。这些结果验证了 COO300 是桃和李研究的有力工具。该网络已更名为 PeachGCN v1.0,执行功能预测分析所需的脚本可在 https://github.com/felipecobos/PeachGCN 上获取。
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引用次数: 0
miR156b-targeted VvSBP8/13 functions downstream of the abscisic acid signal to regulate anthocyanins biosynthesis in grapevine fruit under drought. miR156b 靶向 VvSBP8/13 在脱落酸信号下游发挥作用,调节干旱条件下葡萄果实中花青素的生物合成。
Pub Date : 2024-01-02 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad293
Shuihuan Guo, Meng Zhang, Mingxin Feng, Guipeng Liu, Laurent Torregrosa, Xiaoqing Tao, Ruihua Ren, Yulin Fang, Zhenwen Zhang, Jiangfei Meng, Tengfei Xu

Anthocyanins are the primary color components of grapevine berries and wines. In cultivation practices, a moderate water deficit can promote anthocyanin accumulation in red grape skins. Our previous study showed that abscisic acid (ABA) plays a key role in this process. Herein, we identified a microRNA, vv-miR156b, that is generated in grapevine berries in response to drought stress, along with increasing anthocyanin content and biosynthetic structural gene transcripts. In contrast, vv-miR156b short tandem target mimic (STTM) function-loss callus exhibits the opposite phenotype. Results from in vivo and in vitro experiments revealed that the ABA-signaling-regulated transcription factor VvAREB2 binds directly to the ABA-responsive element (ABRE) of the MIR156b promoter and activates miR156b expression. Furthermore, two miR156b downstream targets, VvSBP8 and VvSBP13, exhibited reduced grape anthocyanin content in their overexpressors but there was a contrary result in their CRISPR-edited lines, the decrease in anthocyanin content was rescued in miR156b and SBP8/13 double overexpressors. We further demonstrated that both VvSBP8 and VvSBP13, encoding transcriptional repressors, displayed sufficient ability to interact with VvMYC1 and VvMYBA1, thereby interfering with MYB-bHLH-WD (MBW) repeat transcriptional complex formation, resulting in the repression of anthocyanin biosynthesis. Our findings demonstrate a direct functional relationship between ABA signaling and the miR156-SBP-MBW complex regulatory module in driving drought-induced anthocyanin accumulation in grape berries.

花青素是葡萄浆果和葡萄酒的主要颜色成分。在栽培实践中,适度缺水可促进红葡萄果皮中花青素的积累。我们之前的研究表明,脱落酸(ABA)在这一过程中起着关键作用。在这里,我们发现了一种微RNA--vv-miR156b,它在葡萄浆果中产生,以应对干旱胁迫,同时增加花青素含量和生物合成结构基因转录本。相比之下,vv-miR156b 短串联靶标模拟(STM)功能缺失胼胝体表现出相反的表型。体内和体外实验结果表明,ABA 信号调控转录因子 VvAREB2 可直接与 MIR156b 启动子的 ABA 响应元件(ABRE)结合,并激活 miR156b 的表达。此外,两个 miR156b 下游靶标 VvSBP8 和 VvSBP13 在它们的过表达株中表现出葡萄花青素含量的降低,但在它们的 CRISPR 编辑株中却出现了相反的结果,花青素含量的降低在 miR156b 和 SBP8/13 双过表达株中得到了挽救。我们进一步证明,编码转录抑制因子的 VvSBP8 和 VvSBP13 都有足够的能力与 VvMYC1 和 VvMYBA1 相互作用,从而干扰 MYB-bHLH-WD (MBW)重复转录复合物的形成,导致花青素生物合成受到抑制。我们的研究结果表明,ABA 信号与 miR156-SBP-MBW 复合物调控模块在驱动干旱诱导的葡萄浆果花青素积累方面存在直接的功能关系。
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引用次数: 0
Tissue culture-free transformation of traditional Chinese medicinal plants with root suckering capability. 具有吸根能力的中药植物的无组织培养转化。
Pub Date : 2023-12-29 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad290
Jinghua Lu, Suhui Lu, Chunli Su, Shuai Deng, Mugui Wang, Huan Tang, Zhunian Wang, Guofu Li, Zhaobo Lang, Jian-Kang Zhu
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引用次数: 0
AlliumDB: a central portal for comparative and functional genomics in Allium. AlliumDB:薤白比较和功能基因组学的中心门户。
Pub Date : 2023-12-29 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad285
Pengtao Yang, Yu Yuan, Chao Yan, Yue Jia, Qi You, Lingling Da, Ao Lou, Bingsheng Lv, Zhonghua Zhang, Yue Liu

The genus Allium belongs to the botanical family Amaryllidaceae and includes economically important crops such as onion, garlic, bunching onion, and leek, used as vegetables, spices, and traditional medicines. The large sizes of Allium genomes hamper the genetic dissection of agronomically important traits and molecular breeding. With the growing accumulation of genomic, resequencing, transcriptome, and phenotypic data, the demand for an integrative Allium database is increasing. Here we present a user-friendly database, AlliumDB (https://allium.qau.edu.cn), as a functional genomics hub integrating public and in-house data. The database contains all currently available nuclear and organelle genomes for Allium species, with genes comprehensively annotated based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, orthology, gene families, protein families (Pfam), and non-coding RNA families (Rfam). Transcriptome and variation profiles are integrated into dynamic visualization tools. We took phenotypic photographs and generated trait records for hundreds of Allium germplasms collected worldwide, which are included in the database. We incorporated JBrowse for the visualization of gene structures, RNA sequencing data, and variation data. Analysis tools such as the basic local alignment search tool (BLAST), sequence fetch, enrichment, and motif analyses are available to explore potential gene functions. This database incorporates comprehensive Allium genotypic and phenotypic datasets. As the community assembles new genomes and generates resequencing data for Allium germplasms, the database will be improved and continuously updated with these multi-omics data and comparative genomic studies. We expect the AlliumDB database to become a key resource for the study of Allium crops.

葱属(Allium)属于植物学天南星科(Amaryllidaceae),包括洋葱、大蒜、束葱和韭菜等具有重要经济价值的作物,可用作蔬菜、香料和传统药物。葱属植物基因组庞大,阻碍了对重要农艺性状的遗传分析和分子育种。随着基因组、重测序、转录组和表型数据的不断积累,对综合性 Allium 数据库的需求日益增加。在此,我们提出了一个用户友好型数据库 AlliumDB (https://allium.qau.edu.cn),作为整合公共数据和内部数据的功能基因组学中心。该数据库包含所有目前可用的薤属物种的核基因组和细胞器基因组,并根据基因本体(GO)和京都基因与基因组百科全书(KEGG)分析、基因选集、基因家族、蛋白质家族(Pfam)和非编码 RNA 家族(Rfam)对基因进行了全面注释。转录组和变异图谱被整合到动态可视化工具中。我们为全球收集的数百种薤白种质拍摄了表型照片,并生成了性状记录,这些记录都包含在数据库中。我们将 JBrowse 用于基因结构、RNA 测序数据和变异数据的可视化。基本局部比对搜索工具(BLAST)、序列提取、富集和主题分析等分析工具可用于探索潜在的基因功能。该数据库包含全面的 Allium 基因型和表型数据集。随着社区为薤白种质组装新基因组并生成重测序数据,该数据库将得到改进,并根据这些多组学数据和比较基因组研究不断更新。我们期待 AlliumDB 数据库成为研究薤白作物的关键资源。
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引用次数: 0
IAA-miR164a-NAC100L1 module mediates symbiotic incompatibility of cucumber/pumpkin grafted seedlings through regulating callose deposition. IAA-miR164a-NAC100L1模块通过调节胼胝质沉积介导黄瓜/南瓜嫁接苗的共生不相容。
Pub Date : 2023-12-29 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad287
Mingzhu Yuan, Tong Jin, Jianqiang Wu, Lan Li, Guangling Chen, Jiaqi Chen, Yu Wang, Jin Sun

Grafting is one of the key technologies to overcome the obstacles of continuous cropping, and improve crop yield and quality. However, the symbiotic incompatibility between rootstock and scion affects the normal growth and development of grafted seedlings after survival. The specific molecular regulation mechanism of graft incompatibility is still largely unclear. In this study, we found that the IAA-miR164a-NAC100L1 module induced callose deposition to mediate the symbiotic incompatibility of cucumber/pumpkin grafted seedlings. The incompatible combination (IG) grafting interface accumulated more callose, and the activity of callose synthase (CmCalS1) and IAA content were significantly higher than in the compatible combination (CG). Treatment with IAA polar transport inhibitor in the root of the IG plants decreased CmCalS activity and callose content. Furthermore, IAA negatively regulated the expression of Cm-miR164a, which directly targeted cleavage of CmNAC100L1. Interestingly, CmNAC100L1 interacted with CmCalS1 to regulate its activity. Further analysis showed that the interaction between CmNAC100L1 and CmCalS1 increased the activity of CmCalS1 in the IG plants but decreased it in the CG plants. Point mutation analysis revealed that threonine at the 57th position of CmCalS1 protein played a critical role to maintain its enzyme activity in the incompatible rootstock. Thus, IAA inhibited the expression of Cm-miR164a to elevate the expression of CmNAC100L1, which promoted CmNAC100L1 interaction with CmCalS1 to enhance CmCalS1 activity, resulting in callose deposition and symbiotic incompatibility of cucumber/pumpkin grafted seedlings.

嫁接是克服连作障碍、提高作物产量和品质的关键技术之一。然而,砧木与接穗之间的共生不相容性会影响嫁接苗成活后的正常生长发育。目前,嫁接不亲和的具体分子调控机制还很不清楚。本研究发现,IAA-miR164a-NAC100L1模块诱导胼胝质沉积介导黄瓜/南瓜嫁接苗的共生不相容。不相容组合(IG)嫁接界面积累了更多的胼胝质,胼胝质合成酶(CmCalS1)的活性和IAA含量明显高于相容组合(CG)。在 IG 植株根部使用 IAA 极性运输抑制剂会降低 CmCalS 的活性和胼胝质含量。此外,IAA 对 Cm-miR164a 的表达有负向调节作用,Cm-miR164a 可直接定向裂解 CmNAC100L1。有趣的是,CmNAC100L1 与 CmCalS1 相互作用以调节其活性。进一步的分析表明,CmNAC100L1与CmCalS1之间的相互作用提高了IG植株中CmCalS1的活性,但降低了CG植株中CmCalS1的活性。点突变分析表明,CmCalS1蛋白第57位的苏氨酸对维持其在不相容砧木中的酶活性起着关键作用。因此,IAA抑制了Cm-miR164a的表达,从而提高了CmNAC100L1的表达,促进了CmNAC100L1与CmCalS1的相互作用,增强了CmCalS1的活性,导致胼胝质沉积和黄瓜/南瓜嫁接苗的共生不相容。
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引用次数: 0
A chromosome-level genome assembly of Agave hybrid NO.11648 provides insights into the CAM photosynthesis. 龙舌兰杂交种 NO.11648 的染色体级基因组组装提供了对 CAM 光合作用的深入了解。
Pub Date : 2023-12-19 eCollection Date: 2024-02-01 DOI: 10.1093/hr/uhad269
Ziping Yang, Qian Yang, Qi Liu, Xiaolong Li, Luli Wang, Yanmei Zhang, Zhi Ke, Zhiwei Lu, Huibang Shen, Junfeng Li, Wenzhao Zhou

The subfamily Agavoideae comprises crassulacean acid metabolism (CAM), C3, and C4 plants with a young age of speciation and slower mutation accumulation, making it a model crop for studying CAM evolution. However, the genetic mechanism underlying CAM evolution remains unclear because of lacking genomic information. This study assembled the genome of Agave hybrid NO.11648, a constitutive CAM plant belonging to subfamily Agavoideae, at the chromosome level using data generated from high-throughput chromosome conformation capture, Nanopore, and Illumina techniques, resulting in 30 pseudo-chromosomes with a size of 4.87 Gb and scaffold N50 of 186.42 Mb. The genome annotation revealed 58 841 protein-coding genes and 76.91% repetitive sequences, with the dominant repetitive sequences being the I-type repeats (Copia and Gypsy accounting for 18.34% and 13.5% of the genome, respectively). Our findings also provide support for a whole genome duplication event in the lineage leading to A. hybrid, which occurred after its divergence from subfamily Asparagoideae. Moreover, we identified a gene duplication event in the phosphoenolpyruvate carboxylase kinase (PEPCK) gene family and revealed that three PEPCK genes (PEPCK3, PEPCK5, and PEPCK12) were involved in the CAM pathway. More importantly, we identified transcription factors enriched in the circadian rhythm, MAPK signaling, and plant hormone signal pathway that regulate the PEPCK3 expression by analysing the transcriptome and using yeast one-hybrid assays. Our results shed light on CAM evolution and offer an essential resource for the molecular breeding program of Agave spp.

姬松茸亚科(Agavoideae)包括茎基酸代谢(CAM)、C3 和 C4 植物,其物种形成的年龄较小,变异积累较慢,因此是研究 CAM 进化的模式作物。然而,由于缺乏基因组信息,CAM进化的遗传机制仍不清楚。本研究利用高通量染色体构象捕获、Nanopore和Illumina技术产生的数据,在染色体水平上组装了龙舌兰杂交种NO.11648的基因组,得到了30条伪染色体,大小为4.87 Gb,支架N50为186.42 Mb。基因组注释发现了 58 841 个蛋白编码基因和 76.91% 的重复序列,其中最主要的重复序列是 I 型重复序列(Copia 和 Gypsy 分别占基因组的 18.34% 和 13.5%)。我们的研究结果还支持在杂交种从天南星亚科(Asparagoideae)分化出来之后,在杂交种的世系中发生了一次全基因组复制事件。此外,我们还发现了磷酸烯醇丙酮酸羧化酶激酶(PEPCK)基因家族的基因复制事件,并揭示了三个 PEPCK 基因(PEPCK3、PEPCK5 和 PEPCK12)参与了 CAM 通路。更重要的是,我们通过分析转录组和使用酵母单杂交实验,确定了富含在昼夜节律、MAPK 信号和植物激素信号通路中调控 PEPCK3 表达的转录因子。我们的研究结果揭示了 CAM 的进化过程,为龙舌兰属植物的分子育种计划提供了重要资源。
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引用次数: 0
A biosynthetic network for protoberberine production in Coptis chinensis. 黄连生产原小檗碱的生物合成网络。
Pub Date : 2023-12-13 eCollection Date: 2024-01-01 DOI: 10.1093/hr/uhad259
Linrui Wu, Binxin Zhao, Zixin Deng, Bin Wang, Yi Yu

Protoberberine alkaloids are a group of tetracyclic isoquinoline compounds known for their well-established antimicrobial and anti-inflammatory properties. The richness and diversity of protoberberine alkaloids accumulated in the Coptis genus necessitate a comprehensive examination of the biosynthetic machinery to understand their ecological significance. Here, from Coptis chinensis we identified CcCYP719A1, which could install a methylenedioxy bridge on either ring A or ring D of the protoberberine backbone, thus diverging metabolite flux towards the biosynthesis of various protoberberine components. We also obtained CcCYP719A2 and CcCYP719A3, which underwent positive selection after diverging from CcCYP719A1 and maintained specific catalytic activity on ring D. Further, we resolved the biosynthetic pathway of jatrorrhizine by identifying two demethylases, which could also modulate protoberberine composition by removing the C-3 methyl group and methylenedioxy bridge of ring D, allowing demethylated metabolites to be redirected into different routes. Moreover, we characterized 2-O-methyltransferase CcOMT1 and flavin-dependent oxidase CcTHBO, respectively responsible for the commonly observed 2-O-methylation and aromatic ring-C assembly in protoberberine alkaloids. Overall, this study reveals an interconnected metabolite network from which diverse protoberberine alkaloids originate. It provides valuable insights into the existence of undiscovered protoberberine components, and paves the way for the targeted production of desired protoberberine components for potential therapeutic development.

原小檗碱是一组四环异喹啉化合物,具有公认的抗菌和消炎特性。由于黄连属积累了丰富多样的原小檗碱生物碱,因此有必要对其生物合成机制进行全面研究,以了解其生态学意义。在这里,我们从黄连中鉴定出了 CcCYP719A1,它可以在原小檗碱骨架的 A 环或 D 环上安装亚甲基二氧桥,从而使各种原小檗碱成分的生物合成代谢通量发生分化。我们还获得了 CcCYP719A2 和 CcCYP719A3,它们在与 CcCYP719A1 分歧后进行了正向选择,并保持了对 D 环的特异性催化活性。此外,我们还通过鉴定两种去甲基酶解决了药根碱的生物合成途径问题,这两种酶也可以通过去除 D 环上的 C-3 甲基和亚甲二氧基桥来调节原小檗碱的组成,从而使去甲基代谢物转向不同的途径。此外,我们还鉴定了 2-O-甲基转移酶 CcOMT1 和黄素依赖性氧化酶 CcTHBO,它们分别负责原小檗碱中常见的 2-O-甲基化和芳香环-C 组装。总之,这项研究揭示了一个相互关联的代谢物网络,各种原小檗碱生物碱就来源于这个网络。它为了解尚未发现的原小檗碱成分的存在提供了有价值的见解,并为有针对性地生产所需的原小檗碱成分以进行潜在的治疗开发铺平了道路。
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引用次数: 0
A highly diversified NLR cluster in melon contains homologs that confer powdery mildew and aphid resistance. 甜瓜中一个高度多样化的 NLR 簇包含赋予白粉病和蚜虫抗性的同源物。
Pub Date : 2023-12-13 eCollection Date: 2024-01-01 DOI: 10.1093/hr/uhad256
Nathalie Boissot, Veronique Chovelon, Vincent Rittener-Ruff, Nathalie Giovinazzo, Pascale Mistral, Michel Pitrat, Myriam Charpentier, Christelle Troadec, Abdelhafid Bendahmane, Catherine Dogimont

Podosphaera xanthii is the main causal agent of powdery mildew (PM) on Cucurbitaceae. In Cucumis melo, the Pm-w resistance gene, which confers resistance to P. xanthii, is located on chromosome 5 in a cluster of nucleotide-binding leucine-rich repeat receptors (NLRs). We used positional cloning and transgenesis, to isolate the Pm-wWMR 29 gene encoding a coiled-coil NLR (CC-NLR). Pm-wWMR 29 conferred high level of resistance to race 1 of PM and intermediate level of resistance to race 3 of PM. Pm-wWMR 29 turned out to be a homolog of the Aphis gossypii resistance gene Vat-1PI 161375. We confirmed that Pm-wWMR 29 did not confer resistance to aphids, while Vat-1PI 161375 did not confer resistance to PM. We showed that both homologs were included in a highly diversified cluster of NLRs, the Vat cluster. Specific Vat-1PI 161375 and Pm-wWMR 29 markers were present in 10% to 13% of 678 accessions representative of wild and cultivated melon types worldwide. Phylogenic reconstruction of 34 protein homologs of Vat-1PI 161375 and Pm-wWMR 29 identified in 24 melon accessions revealed an ancestor with four R65aa-a specific motif in the LRR domain, evolved towards aphid and virus resistance, while an ancestor with five R65aa evolved towards PM resistance. The complexity of the cluster comprising the Vat/Pm-w genes and its diversity in melon suggest that Vat homologs may contribute to the recognition of a broad range of yet to be identified pests and pathogens.

Podosphaera xanthii 是葫芦科植物白粉病(PM)的主要病原菌。在西甜瓜中,Pm-w 抗性基因位于 5 号染色体上的核苷酸结合富亮氨酸重复受体(NLRs)群中,该基因可使西甜瓜对黄粉病菌产生抗性。我们利用定位克隆和转基因技术分离出了 Pm-wWMR 29 基因,该基因编码一个盘绕线圈 NLR(CC-NLR)。Pm-wWMR 29 对 PM 第 1 种族具有高水平的抗性,对 PM 第 3 种族具有中等水平的抗性。Pm-wWMR 29 原来是蚜虫抗性基因 Vat-1PI 161375 的同源物。我们证实,Pm-WWMR 29 并不赋予蚜虫抗性,而 Vat-1PI 161375 也不赋予 PM 抗性。我们发现这两个同源物都包含在一个高度多样化的 NLRs 簇(即 Vat 簇)中。在代表全球野生和栽培甜瓜类型的 678 个品种中,10% 至 13% 的品种存在特异的 Vat-1PI 161375 和 Pm-wWMR 29 标记。对在 24 个甜瓜品种中鉴定出的 34 个 Vat-1PI 161375 和 Pm-wWMR 29 蛋白同源物进行系统发育重建后发现,其祖先具有四个 R65aa--LRR 结构域中的特异图案,进化为抗蚜虫和病毒,而具有五个 R65aa 的祖先则进化为抗 PM。由 Vat/Pm-w 基因组成的基因簇的复杂性及其在甜瓜中的多样性表明,Vat 同源物可能有助于识别各种尚未确定的害虫和病原体。
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园艺研究(英文)
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