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Multi-Gene Identification and Pathogenicity Analysis of Sugarcane Pokkah Boeng Disease Pathogens in Yunnan, China 中国云南甘蔗白粉病病原体的多基因鉴定和致病性分析
IF 2 4区 生物学 Q2 PLANT SCIENCES Pub Date : 2024-09-06 DOI: 10.1007/s12042-024-09368-y
Changmi Wang, Jiong Yin, Yinhu Li, Jie Li, Rongyue Zhang, Xiaoyan Wang, Hongli Shan, Zhiming Luo

Pokkah boeng disease (PBD), a fungal disease of sugarcane, is caused by multiple species of Fusarium. To identify the pathogens causing PBD in Yunnan Province, China, we collected 87 leaf samples with typical PBD symptoms from the sugarcane-growing areas of Puer, Honghe, and Lincang, from which eight strains (FS1–FS8) were isolated and purified. The identities of the eight isolates were determined based on a combination of morphological observations and molecular analyses, using which we identified five Fusarium species as the causal agents of PBD in Yunnan, namely, F. incarnatum, F. andiyazi, F. sacchari, F. proliferatum, and F. verticillioides. Among these, F. incarnatum is a newly recorded species in Yunnan. Pathogenicity tests revealed that all five Fusarium species can infect sugarcane and cause PBD symptoms, with the severity of pathogenicity, ranked from strong to weak, being ordered as follows: F. proliferatum > F. andiyazi > F. incarnatum > F. verticillioides > F. sacchari. In this study, we identified the species and established the pathogenicity of the causal agents of PBD in Yunnan Province. Our findings will provide a scientific basis for the prevention and control of this disease.

Pokkah boeng 病(PBD)是一种甘蔗真菌病害,由多种镰刀菌引起。为了鉴定中国云南省引起 PBD 的病原菌,我们从普洱、红河和临沧的甘蔗种植区采集了 87 份具有典型 PBD 症状的叶片样本,从中分离并纯化了 8 株菌株(FS1-FS8)。通过形态学观察和分子分析,我们确定了云南 PBD 的病原菌为 5 个镰刀菌种,即 F. incarnatum、F. andiyazi、F. sacchari、F. proliferatum 和 F. verticillioides。其中,F. incarnatum 是云南新记录的物种。致病性试验表明,这五种镰刀菌都能侵染甘蔗并引起 PBD 症状,致病性由强到弱依次为F. proliferatum > F. andiyazi > F. incarnatum > F. verticillioides > F. sacchari。在这项研究中,我们确定了云南省 PBD 病原的种类并确定了其致病性。我们的研究结果将为该病的防控提供科学依据。
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引用次数: 0
Genome-wide Identification and Functional Analysis of RNAi Gene Families in Papaya (Carica papaya L.) 木瓜(Carica papaya L.)RNAi 基因家族的全基因组鉴定和功能分析
IF 2 4区 生物学 Q2 PLANT SCIENCES Pub Date : 2024-07-02 DOI: 10.1007/s12042-024-09366-0
Fee Faysal Ahmed, Afsana Yeasmin Mim, Amina Rownaq, Ive Sultana, Anamika Podder, Md. Abdur Rauf Sarkar

RNA silencing plays a direct functional role in gene expression and defense against pathogens during various developmental stages in plants. This silencing process is regulated by the RNA interference (RNAi) pathway, which relies on double-stranded RNA (dsRNA) generated from small RNAs (sRNAs). Three important protein families regulate the functions of sRNAs: Dicer-like (DCLs), Argonautes (AGOs), and RNA-dependent RNA polymerases (RDRs). These major components have not been identified and characterized in papaya. In this study, we identified the RNAi gene families and comprehensively characterized their regulatory functions through a bioinformatics approach in papaya. The papaya genome contains 3 CaDCL, 8 CaAGO and 4 CaRDR genes, which comprise diverse functional regulatory elements compared to their corresponding Arabidopsis AtDCL, AtAGO, and AtRDR genes, indicating their pivotal roles in the RNA silencing mechanism. Phylogenetic tree and multiple sequence analyses reveal that each CaDCL, CaAGO and CaRDR protein clusters and aligns with the corresponding RNAi genes in Arabidopsis. The results from conserved domain, motif, and gene structure analyses indicate a higher level of similarity within the same gene family with some exceptions. The chromosomal and sub-cellular localization of the predicted proteins shows a well distributed pattern. Furthermore, the gene ontology (GO) analysis demonstrates that the candidate RNAi genes are associated with the RNAi silencing mechanism and related pathways. The network and sub-network analyses reveal interactions between various transcription factors (TFs), such as ERF, Dof, MIKC_MADS, NAC, and BBR-BPC families, and the identified RNAi gene families. Additionally, an analysis of cis-acting regulatory elements uncovers light-responsive (LR), stress-responsive (SR), hormone-responsive (HR), and other activities (OT) functions in the candidate CaDCL, CaAGO, and CaRDR genes. Tissue-specific expression patterns of papaya RNAi genes showed differential expression in various tissues. RNA-seq analysis revealed five candidates CpDCL1, CpAGO1, CpAGO5, CpAGO10a, and CpRDR6 that demonstrated potential in response to anthracnose-resistant in papaya. Over all, our findings provide useful information for the genetic improvement of papaya cultivars in breeding programs to overcome stress responses and offer a better understanding of these gene families in papaya.

在植物的各个发育阶段,RNA 沉默在基因表达和抵御病原体方面发挥着直接的功能作用。这一沉默过程受 RNA 干扰(RNAi)途径调控,而 RNA 干扰则依赖于小 RNA(sRNA)产生的双链 RNA(dsRNA)。有三个重要的蛋白家族调节着 sRNAs 的功能:Dicer-like (DCLs)、Argonautes (AGOs) 和 RNA 依赖性 RNA 聚合酶 (RDRs)。在番木瓜中,这些主要成分尚未被鉴定和表征。在这项研究中,我们确定了 RNAi 基因家族,并通过生物信息学方法全面描述了它们在番木瓜中的调控功能。木瓜基因组中含有3个CaDCL、8个CaAGO和4个CaRDR基因,与拟南芥中相应的AtDCL、AtAGO和AtRDR基因相比,木瓜基因组中的CaDCL、CaAGO和CaRDR基因由不同的功能调控元件组成,表明它们在RNA沉默机制中起着关键作用。系统发生树和多重序列分析表明,每种 CaDCL、CaAGO 和 CaRDR 蛋白都与拟南芥中相应的 RNAi 基因聚类和一致。保守结构域、主题和基因结构分析的结果表明,除个别情况外,同一基因家族内的相似度较高。预测蛋白质的染色体和亚细胞定位显示出良好的分布模式。此外,基因本体(GO)分析表明,候选 RNAi 基因与 RNAi 沉默机制及相关途径有关。网络和子网络分析揭示了各种转录因子(TFs),如 ERF、Dof、MIKC_MADS、NAC 和 BBR-BPC 家族与已发现的 RNAi 基因家族之间的相互作用。此外,对顺式作用调控元件的分析揭示了候选 CaDCL、CaAGO 和 CaRDR 基因的光响应(LR)、胁迫响应(SR)、激素响应(HR)和其他活动(OT)功能。木瓜 RNAi 基因的组织特异性表达模式显示了在不同组织中的差异表达。RNA-seq分析揭示了五个候选基因CpDCL1、CpAGO1、CpAGO5、CpAGO10a和CpRDR6,这些基因在应对木瓜抗炭疽病方面具有潜力。总之,我们的研究结果为育种计划中木瓜栽培品种克服胁迫反应的遗传改良提供了有用的信息,并使人们对木瓜中的这些基因家族有了更好的了解。
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引用次数: 0
Identification of novel marker-trait associations and candidate genes for combined low phosphorus and nitrogen-deficient conditions in rice at seedling stage 鉴定水稻幼苗期低磷缺氮联合条件下的新型标记性状关联和候选基因
IF 2 4区 生物学 Q2 PLANT SCIENCES Pub Date : 2024-06-26 DOI: 10.1007/s12042-024-09365-1
Parameswaran Chidambaranathan, Shivraj Sahu, Sabarinathan Selvaraj, Reshmi Raj, Cayalvizhi Balasubramaniasai, Sanghamitra Samantaray, Baishnab Charan Muduli, Anandan Annamalai, Jitendriya Meher, Dibyendu Chatterjee, Sangita Mohanty, Padmini Swain, Lambodar Behera

Rice responds to individual N and P deficiencies through root traits’ modifications and characteristic starvation responses. The genomic regions associated with combined deficiencies of N and P are less reported, though the combinatorial regulation of N and P deficiencies is vital for seedling development. In this study, genome-wide association analysis (GWAS) using ~ 22 k SNPs was performed in one hundred and thirty rice genotypes for nine different traits at the seedling stage (21 days after sowing), and twenty-four statistically significant marker trait associations contributing to the phenotypic variation of 10–79% were identified. Further, except for 10% increase in root length, traits like shoot length, number of leaves, shoot area, shoot dry weight, and root dry weight decreased by 45%, 15%, 60%, 24%, and 45%, respectively, under N and P-deficient soils. Besides, candidate genes for root architecture remodeling (Dro1 and Sor1), P and N uptake (PTF1, PEPC), and amino acid transport and homeostasis (AAP7, BCAT2) were found within the genomic regions regulating the combined tolerance to low P and low N. Furthermore, three superior genotypes, namely ENT-62 (Root area, shoot area, and shoot dry weight), ENT-303 (shoot dry weight and root dry weight), and ENT-32 (no. of leaves and shoot area), were identified for regulating more than one trait under low P-low N conditions. Therefore, this study characterized the seedling stage trait response in rice genotypes and identified genomic regions regulating seedling traits for combined N and P deficient soils. The identified QTLs of these genes could be utilized in breeding programs for the combined improvement of nitrogen and phosphorus use efficiency under deficit soils.

水稻通过根系性状的改变和特有的饥饿反应来应对单个氮和磷的缺乏。尽管氮和磷的综合调控对幼苗的生长发育至关重要,但与氮和磷的综合缺乏相关的基因组区域却鲜有报道。本研究利用约 22 k 个 SNPs 对 130 个水稻基因型的幼苗期(播种后 21 天)的 9 个不同性状进行了全基因组关联分析(GWAS),发现了 24 个对表型变异有 10-79% 贡献的具有统计学意义的标记性状关联。此外,在缺氮和缺磷的土壤中,除了根长增加了 10%外,芽长、叶片数、芽面积、芽干重和根干重等性状分别减少了 45%、15%、60%、24% 和 45%。此外,在调控根系结构重塑(Dro1 和 Sor1)、P 和 N 吸收(PTF1 和 PEPC)以及氨基酸转运和稳态(AAP7 和 BCAT2)的基因组区域内发现了候选基因。此外,在低磷低氮条件下,ENT-62(根面积、芽面积和芽干重)、ENT-303(芽干重和根干重)和ENT-32(叶片数和芽面积)这三个优良基因型可调控一个以上的性状。因此,本研究描述了水稻基因型的苗期性状响应,并确定了在缺氮和缺磷的综合土壤条件下调控苗期性状的基因组区域。这些基因的 QTLs 可用于育种计划,以综合提高缺氮和缺磷土壤的氮磷利用效率。
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引用次数: 0
Comprehensive Analysis of the Aquaporin Genes in Eucalyptus grandis Suggests Potential Targets for Drought Stress Tolerance 全面分析大桉树的水蒸气素基因,为耐干旱胁迫提供潜在靶标
IF 2 4区 生物学 Q2 PLANT SCIENCES Pub Date : 2024-06-14 DOI: 10.1007/s12042-024-09364-2
Dayana S. Seidel, Paulo H. Claudino, Gabriela Sperotto, Simone N. Wendt, Zachery D. Shomo, Ravi V. Mural, Henrique M. Dias

This study delves into the comprehensive analysis of AQP genes in Eucalyptus grandis, providing insights into their genomic abundance, diversification, expression patterns across tissues, and responses to drought stress. We identified 48 AQP genes in the Eucalyptus grandis genome, categorized into four subfamilies: AQP-NIP, AQP-SIP, AQP-PIP, and AQP-TIP. This abundance of AQP genes is a reflection of gene duplications, both tandem and whole-genome, which have shaped their expansion. The chromosomal distribution of these genes reveals their widespread presence across the genome, with some subfamilies exhibiting more tandem duplications, suggesting distinct roles and evolutionary pressures. Sequence analysis uncovered characteristic motifs specific to different AQP subfamilies, demonstrating the diversification of protein and targeting. The expression profiles of AQP genes in various tissues in both Arabidopsis thaliana and Eucalyptus grandis showcased variations, with root tissues showing higher expression levels. Notably, AQP-PIP genes consistently exhibited robust expression across tissues, highlighting their importance in maintaining water regulation within plants. Furthermore, the study investigated the response of AQP genes to drought stress and rehydration, revealing differential expression patterns. EgAQP-NIP and EgAQP-TIP genes were up-regulated during drought stress, emphasizing their role in osmotic equilibrium and water transport. Conversely, EgAQP-PIP genes showed down-regulation during drought stress but were up-regulated upon rehydration, indicating their involvement in water movement across cell membranes. Overall, this research contributes to our understanding of AQP genes in Eucalyptus grandis, shedding light on their genomic evolution, expression patterns, and responses to environmental challenges, particularly drought stress. This information can be valuable for future studies aimed at enhancing the drought resilience of woody perennial plants like Eucalyptus grandis.

本研究对桉树中的 AQP 基因进行了全面分析,深入了解了这些基因的基因组丰度、多样性、跨组织表达模式以及对干旱胁迫的响应。我们在桉树基因组中发现了 48 个 AQP 基因,分为四个亚家族:AQP-NIP、AQP-SIP、AQP-PIP 和 AQP-TIP。大量的 AQP 基因反映了基因的重复,包括串联重复和全基因组重复,这些重复形成了 AQP 基因的扩展。这些基因在染色体上的分布显示了它们在整个基因组中的广泛存在,其中一些亚家族出现了更多的串联重复,这表明它们具有不同的作用和进化压力。序列分析发现了不同 AQP 亚家族特有的特征基调,证明了蛋白质和靶向的多样化。拟南芥和桉树的 AQP 基因在不同组织中的表达谱呈现出差异,根部组织的表达水平较高。值得注意的是,AQP-PIP 基因在不同组织中始终表现出较强的表达能力,这凸显了它们在植物体内维持水分调节的重要性。此外,研究还调查了 AQP 基因对干旱胁迫和补水的响应,发现了不同的表达模式。在干旱胁迫期间,EgAQP-NIP 和 EgAQP-TIP 基因上调,强调了它们在渗透平衡和水分运输中的作用。相反,EgAQP-PIP 基因在干旱胁迫期间下调,但在复水后上调,表明它们参与了细胞膜上的水分运输。总之,这项研究有助于我们了解桉树的 AQP 基因,揭示它们的基因组进化、表达模式以及对环境挑战(尤其是干旱胁迫)的反应。这些信息对今后旨在提高桉树等多年生木本植物抗旱能力的研究很有价值。
{"title":"Comprehensive Analysis of the Aquaporin Genes in Eucalyptus grandis Suggests Potential Targets for Drought Stress Tolerance","authors":"Dayana S. Seidel, Paulo H. Claudino, Gabriela Sperotto, Simone N. Wendt, Zachery D. Shomo, Ravi V. Mural, Henrique M. Dias","doi":"10.1007/s12042-024-09364-2","DOIUrl":"https://doi.org/10.1007/s12042-024-09364-2","url":null,"abstract":"<p>This study delves into the comprehensive analysis of <i>AQP</i> genes in <i>Eucalyptus grandis</i>, providing insights into their genomic abundance, diversification, expression patterns across tissues, and responses to drought stress. We identified 48 <i>AQP</i> genes in the <i>Eucalyptus grandis</i> genome, categorized into four subfamilies: <i>AQP-NIP</i>, <i>AQP-SIP</i>, <i>AQP-PIP</i>, and <i>AQP-TIP</i>. This abundance of <i>AQP</i> genes is a reflection of gene duplications, both tandem and whole-genome, which have shaped their expansion. The chromosomal distribution of these genes reveals their widespread presence across the genome, with some subfamilies exhibiting more tandem duplications, suggesting distinct roles and evolutionary pressures. Sequence analysis uncovered characteristic motifs specific to different AQP subfamilies, demonstrating the diversification of protein and targeting. The expression profiles of <i>AQP</i> genes in various tissues in both <i>Arabidopsis thaliana</i> and <i>Eucalyptus grandis</i> showcased variations, with root tissues showing higher expression levels. Notably, <i>AQP-PIP</i> genes consistently exhibited robust expression across tissues, highlighting their importance in maintaining water regulation within plants. Furthermore, the study investigated the response of <i>AQP</i> genes to drought stress and rehydration, revealing differential expression patterns. <i>EgAQP-NIP</i> and <i>EgAQP-TIP</i> genes were up-regulated during drought stress, emphasizing their role in osmotic equilibrium and water transport. Conversely, <i>EgAQP-PIP</i> genes showed down-regulation during drought stress but were up-regulated upon rehydration, indicating their involvement in water movement across cell membranes. Overall, this research contributes to our understanding of <i>AQP</i> genes in <i>Eucalyptus grandis</i>, shedding light on their genomic evolution, expression patterns, and responses to environmental challenges, particularly drought stress. This information can be valuable for future studies aimed at enhancing the drought resilience of woody perennial plants like <i>Eucalyptus grandis</i>.</p>","PeriodicalId":54356,"journal":{"name":"Tropical Plant Biology","volume":null,"pages":null},"PeriodicalIF":2.0,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141532433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Genome-Wide Identification and Expression Analysis of WRKY Transcription Factor Genes in Passion Fruit (Passiflora edulis) 百香果(西番莲)中 WRKY 转录因子基因的全基因组鉴定和表达分析
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-05-31 DOI: 10.1007/s12042-024-09355-3
Shugang Xu, Xingcheng Zhu, Qian Zhang, Shixian Zeng, Yan Li, Yong Wang

The WRKY gene family is an important class of transcription factors in higher plants that play key roles in secondary metabolism, phytohormone signaling, plant defense responses, and abiotic stress responses. The WRKY gene family has not been systematically studied in Passiflora edulis, a tropical fruit with edible and medicinal values. In this study, we performed a genome-wide analysis of passion fruit and identified 58 candidate PeWRKY genes distributed unevenly on nine chromosomes of passion fruit. Phylogenetic and gene structure analyses showed that members of the PeWRKY gene family could be categorized into three groups: class I (9), class II (40), and class III (9). Promoter and target gene prediction analyses indicated that the PeWRKY gene may be involved in various biological processes, including growth and development, metabolism, hormones, and stress responses, by regulating multiple target genes. The Ka/Ks ratios of PeWRKY indicated that PeWRKY may have undergone strong purification selection in the passion fruit genome. Tissue expression analysis showed that the PeWRKY gene was expressed in different tissues of Passiflora edulis, where the number and expression of the PeWRKY gene were more significant in the roots. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that the relative expression of all 10 candidate PeWRKY genes was significantly up-regulated in leaf tissues after 48 h of drought stress compared with the control, and only PeWRKY55 and PeWRKY30 genes were down-regulated. In addition, at the time of biotic stress treatment up to 24 h, most PeWRKY genes were up-regulated after treatment, except the PeWRKY40 gene, which showed down-regulation. Three genes, PeWRKY02, PeWRKY30 and PeWRKY58, showed significant up-regulation of their expression after treatment up to 48 h. This study provides a valuable reference for the functional characterization of WRKY genes in passion fruit and other plants.

WRKY 基因家族是高等植物中一类重要的转录因子,在次生代谢、植物激素信号转导、植物防御反应和非生物胁迫反应中发挥关键作用。西番莲是一种具有食用和药用价值的热带水果,目前尚未对其 WRKY 基因家族进行系统研究。在这项研究中,我们对百香果进行了全基因组分析,发现了 58 个候选 PeWRKY 基因,它们不均匀地分布在百香果的 9 条染色体上。系统发育和基因结构分析表明,PeWRKY 基因家族成员可分为三类:I 类(9 个)、II 类(40 个)和 III 类(9 个)。启动子和靶基因预测分析表明,PeWRKY 基因可能通过调控多个靶基因参与多种生物过程,包括生长发育、新陈代谢、激素和应激反应。PeWRKY 的 Ka/Ks 比值表明,PeWRKY 可能在百香果基因组中经历了强烈的纯化选择。组织表达分析表明,PeWRKY基因在西番莲的不同组织中均有表达,其中根部的PeWRKY基因数量和表达量更为显著。定量实时聚合酶链反应(qRT-PCR)显示,与对照相比,干旱胁迫 48 h 后叶片组织中所有 10 个候选 PeWRKY 基因的相对表达量均显著上调,只有 PeWRKY55 和 PeWRKY30 基因下调。此外,在生物胁迫处理 24 h 内,除 PeWRKY40 基因下调外,大多数 PeWRKY 基因在处理后上调。该研究为百香果及其他植物中 WRKY 基因的功能表征提供了有价值的参考。
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引用次数: 0
Molecular Cloning and Functional Analysis of ScHAK10 Gene Promoter from Sugarcane (Saccharum officinarum L.) 甘蔗(Saccharum officinarum L.)SCHAK10 基因启动子的分子克隆和功能分析
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-05-17 DOI: 10.1007/s12042-024-09363-3
Hai-Bin Luo, Cheng-Mei Huang, Hui-Qing Cao, Yuan-Wen Wei, Lin Xu, Kai-Chao Wu, Zhi-Nian Deng, Xing-Jian Wu, Li-Ping Ye, Xiao-Ping Yi
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引用次数: 0
Silencing of Coat Protein Gene Using IhpRNA Develops Resistance to Banana Bract Mosaic Virus in Musa Acuminata (AAA) cv. Grand Naine 使用 IhpRNA 沉默鞘蛋白基因可提高香蕉苞片花叶病毒在钝茎麝香(AAA)变种中的抗性Grand Naine
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-05-11 DOI: 10.1007/s12042-024-09360-6
Pritam Ramesh Jadhav, Sachin Chandrakant Ekatpure, K. B. Soni, Alex Swapna, R. S. Lekshmi, Yogesh Sahebrao Wagh, R. V. Manju

Banana bract mosaic virus (BBrMV), transmitted by aphids, is a major threat to banana cultivation, causing substantial economic losses. This study focuses on the development of BBrMV-resistant lines of banana cv. ‘Grand Naine’ by silencing viral coat protein (CP) gene using RNA interference (RNAi) strategy. To achieve this, an intron hairpin RNA (ihpRNA) construct containing a 326 bp fragment of the CP gene was designed using the pSTARLING vector. Identification of a Dicer substrate within the CP gene facilitated the prediction of small interfering RNAs (siRNAs) through Custom Dicer-Substrate siRNA analysis. The absence of viral silencing suppressors was validated using the VsupPred tool. Cloning of the sense and antisense fragments of the CP gene into the pSTARLING vector, flanking the cre intron, was confirmed through PCR analysis. Subsequently, the NotI fragment comprising the ubiquitin promoter, ubiquitin intron, sense fragment inserts, cre intron, antisense strand insert, and tumour morphology locus (tmL) terminator was transferred to the Agrobacterium tumefaciens binary vector pART27. Embryogenic calli were transformed with the ihpRNA-CP cassette, and regenerated plantlets were screened for complete cassette integration using PCR. Northern hybridization confirmed the production of siRNAs against coat protein mRNA. Upon exposure to virulent aphids carrying BBrMV, the transformed lines exhibited no disease symptoms. Additionally, reverse transcription quantitative PCR (RT-qPCR) demonstrated the absence of BBrMV, with transformed lines resembling healthy, non-inoculated controls both morphologically and in terms of coat protein gene expression. This RNAi-based approach showcases the successful creation of BBrMV-resistant banana lines, presenting a promising strategy for combating the virus's detrimental effects on banana cultivation.

由蚜虫传播的香蕉苞片花叶病毒(BBrMV)是香蕉种植的主要威胁,造成了巨大的经济损失。本研究的重点是利用 RNA 干扰(RNAi)策略沉默病毒衣壳蛋白(CP)基因,培育香蕉品种 "Grand Naine "的 BBrMV 抗性品系。为此,使用 pSTARLING 载体设计了一个内含子发夹 RNA(ihpRNA)构建体,其中包含 CP 基因的 326 bp 片段。CP 基因中 Dicer 底物的鉴定有助于通过定制 Dicer 底物 siRNA 分析预测小干扰 RNA(siRNA)。使用 VsupPred 工具验证了病毒沉默抑制因子的缺失。通过 PCR 分析确认了将 CP 基因的有义和反义片段克隆到 pSTARLING 载体中,侧翼为 cre 内含子。随后,由泛素启动子、泛素内含子、有意义片段插入物、cre 内含子、反义链插入物和肿瘤形态基因座(tmL)终止子组成的 NotI 片段被转入农杆菌双元载体 pART27。用ihpRNA-CP基因盒转化胚胎胼胝体,并用PCR筛选再生的小植株是否完全整合了基因盒。Northern 杂交证实产生了针对外壳蛋白 mRNA 的 siRNA。在接触携带 BBrMV 的烈性蚜虫时,转化品系没有表现出疾病症状。此外,反转录定量 PCR(RT-qPCR)证明没有 BBrMV,转化品系在形态和外壳蛋白基因表达方面都与健康的非接种对照相似。这种基于 RNAi 的方法成功培育出了抗 BBrMV 的香蕉品系,为消除病毒对香蕉种植的有害影响提供了一种前景广阔的策略。
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引用次数: 0
Identification and Expression Analysis of Soybean (Glycine max L.) Dynamin Genes Reveal Their Involvements in Plant Development and Stress Response 大豆(Glycine max L.)Dynamin 基因的鉴定和表达分析揭示了它们在植物发育和应激反应中的作用
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-05-04 DOI: 10.1007/s12042-024-09361-5
Xiangbo Duan, Yanang Xu, Ke Zhang, Zhouli Liu, Yang Yu

Dynamin and dynamin-related proteins (DRPs) are large GTPases that are vital for cytokinesis, endocytosis and multiple biological processes. However, knowledge of the DRP gene family in soybean (Glycine max L.), an important leguminous crop plant, is still limited. In this study, 31 GmDRPs were identified from soybean genome, and were classified into five groups based on phylogenetic analysis. We observed that each group displayed specific conserved domain distribution and exon–intron structures. Collinearity analysis indicated that gene duplication events contribute largely to the expansion of GmDRP family. According to functional annotation, soybean dynamins were found implicated in cell division, endocytosis, and mitochondrion/peroxisome fission in GTP-dependent manner. Promoter analysis implied the potential roles of GmDRPs in mediating developmental processes, plant hormone signaling, and stress responses. Based on RNA-seq data, some of the GmDRPs were found ubiquitously expressed in various tissues/organs, some were barely expressed, while some showed obvious tissue/organ-preference. The expression analysis also revealed the involvement of GmDRPs in cold and/or drought stress response. In sum, we performed a systematic analysis of soybean dynamin family and our results provide a foundation for further researches on their functional roles.

动态蛋白和动态蛋白相关蛋白(Dynamin-related proteins,DRPs)是一种大型 GTP 酶,对细胞分裂、内吞和多种生物过程至关重要。然而,人们对大豆(Glycine max L.)这一重要豆科作物中 DRP 基因家族的了解仍然有限。本研究从大豆基因组中鉴定了 31 个 GmDRPs,并根据系统发育分析将其分为五组。我们观察到每个组都显示出特定的保守结构域分布和外显子-内含子结构。共线性分析表明,基因复制事件在很大程度上导致了 GmDRP 家族的扩展。根据功能注释,大豆动态蛋白以 GTP 依赖性方式参与细胞分裂、内吞和线粒体/过氧物酶体裂变。启动子分析表明,GmDRPs 在介导发育过程、植物激素信号转导和胁迫响应方面具有潜在作用。根据 RNA-seq 数据,发现一些 GmDRPs 在不同组织/器官中普遍表达,一些几乎不表达,而另一些则表现出明显的组织/器官偏好。表达分析还揭示了 GmDRPs 在冷和/或干旱胁迫响应中的参与。总之,我们对大豆动态蛋白家族进行了系统分析,我们的研究结果为进一步研究其功能作用奠定了基础。
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引用次数: 0
Genome-wide Identification and Characterization of the Ascorbate Peroxidase Gene Family in Citrus sinensis in Response to Huanglongbing 应对黄龙病的柑橘抗坏血酸过氧化物酶基因家族的全基因组鉴定和特征描述
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-05-03 DOI: 10.1007/s12042-024-09362-4
Ruimin Li, Cheng Yang, Xinyou Wang, Yana Yan, Guiyan Huang

Ascorbate peroxidases (APXs) are essential for plants as they act as hydrogen peroxide-scavenging enzymes, providing protection against oxidative damage. Using bioinformatic methods, five APX genes were discovered in the genome of Citrus sinensis in this study. APX genes of C. sinensis (CsAPXs) encode polypeptides between 250 and 436 residues in length, with molecular weights that range from 27.56 to 47.34 kDa. Additionally, the isoelectric point of CsAPXs varies from 5.64 to 8.63. The predicted locations of CsAPXs are peroxisome, chloroplast, and mitochondrion, with an uneven distribution across four chromosomes and eight orthologous gene pairs with Arabidopsis thaliana. A phylogenetic analysis revealed that the CsAPXs were divided into three clades. The CsAPXs all contained a conserved APX domain and six common motifs. Upon promoter analysis, it was found that CsAPXs could respond to abscisic acid, auxin, ethylene, gibberellin, methyl jasmonate, salicylic acid, and wounding stress treatments. In addition, the analysis of expression patterns revealed that the presence of Candidatus Liberibacter asiaticus (CLas) has a dynamic impact on the expression of CsAPXs, with CsAPX2 showing significant inhibition in response to CLas infection. These findings will provide novel insights for the forthcoming functional investigations of CsAPXs within the process of citrus-CLas interactions.

抗坏血酸过氧化物酶(APXs)是植物所必需的,因为它们是过氧化氢清除酶,能保护植物免受氧化损伤。本研究利用生物信息学方法,在中华柚基因组中发现了五个 APX 基因。中华芸香科植物的 APX 基因(CsAPXs)编码长度在 250 至 436 个残基之间的多肽,分子量在 27.56 至 47.34 kDa 之间。此外,CsAPXs 的等电点从 5.64 到 8.63 不等。CsAPXs 的预测位置为过氧物酶体、叶绿体和线粒体,在四条染色体上分布不均,与拟南芥有八个同源基因对。系统进化分析表明,CsAPXs 被分为三个支系。CsAPXs 都含有一个保守的 APX 结构域和六个共同的基序。启动子分析发现,CsAPXs 能对脱落酸、辅助素、乙烯、赤霉素、茉莉酸甲酯、水杨酸和伤口胁迫处理做出反应。此外,对表达模式的分析表明,Candidatus Liberibacter asiaticus(CLas)的存在对 CsAPXs 的表达有动态影响,其中 CsAPX2 对 CLas 感染有明显的抑制作用。这些发现将为即将开展的柑橘-CLas 相互作用过程中 CsAPXs 的功能研究提供新的见解。
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引用次数: 0
A Wild Arachis Endochitinase Enhances Sclerotinia Resistance in Transgenic Plants 野生花生内切酶能增强转基因植物对硬菌的抗性
IF 2 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2024-04-12 DOI: 10.1007/s12042-024-09359-z
Deziany da Silva Ferreira, Andressa da Cunha Quintana Martins, Pedro Souza Berbert, Renan Miguel dos Anjos, Mario Alfredo de Passos Saraiva, Ana Cristina Miranda Brasileiro, Robert Neil Gerard Miller, Patricia Messenberg Guimaraes

Plant endochitinases promote the cleavage of chitin, a polymer naturally found in the cell walls of fungi and insects. Although such enzymes are widely employed in plant genetic engineering to increase tolerance to pathogenic fungi, endochitinases from wild germplasm are poorly exploited for biotechnological purposes. Wild peanut species (Arachis spp.) have evolved under a range of environmental conditions and display distinct defensive adaptations, harboring high levels of genetic diversity and constituting an attractive source of resistance genes against pathogens. Arachis stenosperma shows broad resistance against various biotic stresses such as nematodes, fungi, and viruses. Previous transcriptome and proteomic studies on A. stenosperma challenged with fungi and nematodes identified differentially expressed genes (DEGs) involved in plant defense responses, including an upregulated endochitinase (AsECHI1). Here, we characterized endochitinases from 12 different legumes, including wild Arachis species, and evaluated the effects of overexpression of AsECHI1 for control of Sclerotinia sclerotiorum in tobacco, singly and in association with an expansin-like B defense-priming gene (AdEXLB8). Both singly and pyramided transgenic tobacco lines overexpressing AsECHI1 exhibited a delay in disease progression, and up to a 46% reduction in fungal lesions. Further analysis of transgenic plants showed that the overexpression of AsECHI1 led to an increased expression of defense-related genes in the jasmonic acid, auxin, and ethylene biosynthesis pathways, as well as a substantial accumulation of H2O2. These results suggest that the AsECHI1 gene isolated from wild Arachis has the potential to enhance resistance against this highly damaging necrotrophic fungal pathogen, reducing environmental damage related to the use of fungicides and increasing crop sustainability.

植物内几丁质酶能促进几丁质的裂解,几丁质是真菌和昆虫细胞壁中天然存在的聚合物。虽然这类酶被广泛用于植物基因工程,以提高对病原真菌的耐受性,但野生种质中的内几丁质酶却很少被用于生物技术目的。野生花生物种(Arachis spp.)在一系列环境条件下进化,显示出独特的防御适应性,具有高度的遗传多样性,是抗病原体基因的诱人来源。Arachis stenosperma 对线虫、真菌和病毒等各种生物胁迫具有广泛的抗性。之前对受到真菌和线虫挑战的 A. stenosperma 进行的转录组和蛋白质组研究发现了参与植物防御反应的差异表达基因(DEGs),包括上调的内吸素酶(AsECHI1)。在这里,我们鉴定了来自 12 种不同豆科植物(包括野生Arachis物种)的内切酶的特征,并评估了单独过表达AsECHI1和与扩张素样B防御激发基因(AdEXLB8)联合过表达AsECHI1对烟草中硬皮病的控制效果。过量表达 AsECHI1 的单一转基因烟草品系和金字塔转基因烟草品系都能延缓病害的发展,真菌病害最多可减少 46%。对转基因植株的进一步分析表明,AsECHI1 的过表达导致茉莉酸、辅助素和乙烯生物合成途径中防御相关基因的表达增加,以及 H2O2 的大量积累。这些结果表明,从野生金花菜中分离出的 AsECHI1 基因有可能增强金花菜对这种破坏性极强的坏死性真菌病原体的抗性,从而减少与使用杀菌剂有关的环境破坏,提高作物的可持续发展能力。
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引用次数: 0
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Tropical Plant Biology
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