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Identification and characterization of ClAPRR2, a key candidate gene controlling watermelon stripe color 西瓜条纹色关键候选基因ClAPRR2的鉴定与特性研究
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-01-03 DOI: 10.1016/j.plantsci.2024.112383
Shuang Liang , Miaomiao Yang , Linlin Zhang , Xufeng Fang , Xian Zhang , Chunhua Wei , Zuyun Dai , Zhongzhou Yang , Chaonan Wang , Bin Liu , Feishi Luan , Shi Liu
The stripe color of watermelon is a vital commercial trait and is the focus of attention of consumers and researchers. However, the genetic determinants of watermelon stripe color are incompletely understood. Based on the results of preliminary localization studies, we constructed a large-capacity F2 generation population (710 plants) using light-green striped ZXG1555 and green-striped Cream of Saskatchewan (COS) watermelon strains as parental lines for fine mapping. Genes controlling stripe color were located in an 85.284 kb region on chromosome 9, which contained five candidate genes. Combined with parental phenotypes, chlorophyll contents of rinds and stripes were assayed. Gene sequence alignment and transcriptional level analysis of parental lines predicted Cla97C09G175170 (encoding a two-component response regulator-like protein, APRR2) as the best candidate gene for stripe color trait. Two SNPs in the ClAPRR2 coding region caused amino acid substitutions, but were not located in the conserved domain, while a 12 bp insertion caused premature translation termination and a 35 amino acid deletion in the conserved domain and may have affected ClAPRR2 function in ZXG1555. Subcellular localization analysis showed that ClAPRR2 was expressed in the ZXG1555 cell membrane but was located in the nucleus and cell membrane of COS. Nucleotide polymorphisms and deletions were also detected in the promoter region between parental lines and caused cis-acting element variations. Luciferase activity suggested that promoter variations may not be the main factor in the regulation of ClAPRR2 expression.
西瓜的条纹颜色是一种重要的商业性状,是消费者和研究人员关注的焦点。然而,西瓜条纹颜色的遗传决定因素尚不完全清楚。在初步定位研究的基础上,以浅绿色条纹的ZXG1555和绿色条纹的Saskatchewan (COS)西瓜品系为亲本,构建了大容量F2代群体(710株)进行精细定位。控制条纹颜色的基因位于第9染色体的85.284kb区域,包含5个候选基因。结合亲本表型,测定果皮和条纹的叶绿素含量。亲本序列比对和转录水平分析预测cl97c09g175170(编码双组分响应调控因子样蛋白APRR2)是条纹色性状的最佳候选基因。ClAPRR2编码区的两个snp导致了氨基酸替换,但并不位于保守结构域,而一个12bp的插入导致了翻译的过早终止和保守结构域35个氨基酸的缺失,这可能影响了ZXG1555的ClAPRR2功能。亚细胞定位分析表明,ClAPRR2在ZXG1555细胞膜中表达,而位于COS的细胞核和细胞膜中。在亲本系之间的启动子区域也检测到核苷酸多态性和缺失,并导致顺式作用元件的变异。荧光素酶活性提示启动子变异可能不是调控ClAPRR2表达的主要因素。
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引用次数: 0
New insights of advanced biotechnological engineering strategies for tanshinone biosynthesis in Salvia miltiorrhiza 丹参酮生物合成先进生物技术工程策略的新见解。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-01-03 DOI: 10.1016/j.plantsci.2025.112384
Minyu Shou , Qinzhe Lin , Ying Xu , Ruiyan Zhu , Min Shi , Guoyin Kai
Salvia miltiorrhiza Bunge, a well-known traditional Chinese herbal medicine, has been served as not only medicine for human ailments, but also health care products. As one of major bioactive ingredients, tanshinones are widely used to treat cardiovascular and cerebrovascular diseases, and also possess different pharmacological activities including anti-tumor, anti-inflammatory, anti-fibrotic and others. However, the content of tanshinones is relatively low in S. miltiorrhiza plants. Recently, multiple biotechnological strategies have been applied to improve tanshinone production. In this review, advances in bioactivities, biosynthesis pathway and regulation, transcriptional regulatory network, epigenetic modification and synthetic biology are summarized, and future perspectives are discussed, which will help develop high-quality S. miltiorrhiza resources.
丹参是一种著名的传统中草药,它不仅是治疗人类疾病的药物,而且是保健品。丹参酮作为主要生物活性成分之一,被广泛用于治疗心脑血管疾病,并具有抗肿瘤、抗炎、抗纤维化等不同的药理活性。丹参酮在丹参中含量相对较低。近年来,多种生物技术策略被应用于提高丹参酮的产量。本文综述了在生物活性、生物合成途径与调控、转录调控网络、表观遗传修饰和合成生物学等方面的研究进展,并对未来的研究方向进行了展望,以期为开发优质的丹参资源提供参考。
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引用次数: 0
Dissecting sequence-structure-function-diversity in plant cryptochromes 植物隐色素序列-结构-功能-多样性分析。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-31 DOI: 10.1016/j.plantsci.2024.112381
Pratichi Sarkar , Aparna Boral , Devrani Mitra
Ubiquitous to every stratum of life, cryptochromes regulate numerous light dependent functions in terrestrial plants. These include light-dependent transcription, circadian rhythm, inhibition of hypocotyl elongation, programmed cell death, promotion of floral initiation, mediation of gravitropic response, responding to biotic and abiotic stress etc. There have been quite a few seminal reviews including on plant cryptochromes, focusing mostly on the detailed functional aspects. This review primarily focuses on understanding the link connecting sequence-structure hierarchy behind the functional diversity in plant cryptochromes. With available sequence information and 3D structure data, we hereby explore the molecular origin of functional diversity in both the subtypes i.e., CRY1 and CRY2. First, we discuss the structural details and functional distinctiveness of all subtypes of plant cryptochromes. Next we draw a comparison not just between two cryptochromes but also other Cryptochrome/Photolyase Family (CPF) members e.g. CRY-DASH/CRY3 and CPD/6–4 photolyases of plant origin. Further, by constructing a phylogenetic profile from multiple sequence alignment we investigate how a crucial activity like DNA repair is restricted to some members of CPF and not all. It is a well-known fact that the function of a protein is heavily if not solely guided by the structure-sequence relationship. Therefore, the resultant hypothesis as drawn from this comparative and collective study could predict functions of many under-studied plant cryptochromes when compared with their well-studied counterparts like Arabidopsis cryptochromes. An extensive sequence-structure-function analysis complemented with evolutionary studies and bibliographic survey is useful towards understanding the immensely diverse CPF.
隐色素普遍存在于生命的各个阶层,在陆生植物中调节着许多依赖光的功能。这些包括光依赖性转录、昼夜节律、抑制下胚轴伸长、细胞程序性死亡、促进花的形成、介导向地性反应、对生物和非生物胁迫的响应等。关于植物隐色素的研究已经有相当多的开创性的综述,主要集中在详细的功能方面。本文综述了植物隐色素功能多样性背后的序列-结构层次关系。利用现有的序列信息和三维结构数据,我们在此探索CRY1和CRY2亚型功能多样性的分子起源。首先,我们讨论了植物隐色素所有亚型的结构细节和功能特征。接下来,我们不仅比较了两种隐色素,还比较了植物来源的其他隐色素/光解酶家族成员,如CRY-DASH/CRY3和CPD/6-4光解酶。此外,通过构建多序列比对的系统发育图谱,我们研究了DNA修复等关键活动如何局限于CPF的某些成员而不是全部成员。这是一个众所周知的事实,蛋白质的功能很大程度上,如果不是完全由结构-序列关系指导。因此,通过比较和集体研究得出的假设可以预测许多研究不足的植物隐色素的功能,并将其与研究充分的同类植物(如拟南芥隐色素)进行比较。广泛的序列-结构-功能分析与进化研究和文献调查相辅相成,有助于理解极其多样化的CPF。
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引用次数: 0
Exploration of advanced omics tools and resources for the improvement of industrial oil crops 经济油料作物改良的先进组学工具和资源探索。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-31 DOI: 10.1016/j.plantsci.2024.112367
Badal Mahakalkar , Virender Kumar , Sreeja Sudhakaran , Vandana Thakral , Sanskriti Vats , Rushil Mandlik , Rupesh Deshmukh , Tilak Raj Sharma , Humira Sonah
The rapid advancement in the field of omics approaches plays a crucial role in the development of improved industrial oil crops. Industrial oil crops are important for many sectors like food processing, biofuels, cosmetics, and pharmaceuticals, making them indispensable contributors to global economies and these crops serve as vital elements in a multitude of industrial processes. Significant improvements in genomics have revolutionized the agricultural sector, particularly in the realm of oil crops. Cutting-edge advancements have facilitated the efficient sequencing of genomes for key commercial oil crops. This breakthrough not only enhances our understanding of the genetic makeup of these crops but also empowers breeders with invaluable insights for targeted genetic manipulation and breeding programs. Moreover, integrating transcriptomics with genomic data has assisted in a new era of precision agriculture. This approach provides an in-depth understanding of molecular mechanisms involved in traits of interest, such as oil content, yield potential, and resistance to biotic and abiotic stresses. Proteomics methods are instrumental in deciphering the intricacies of protein structure, interactions, and function, while metabolomics and ionomics shed light on the intricate network of metabolites and ions within biological systems. Each omics discipline offers unique insights, and their integration holds the promise of enriching our understanding and furnishing invaluable insights for enhancing oil crops. This review delves into the efficacy and constraints of various omics approaches in the context of refining industrial oil crops. Moreover, it underscores the importance of multi-omics strategies and explores their convergence with genetic engineering techniques to cultivate superior oil crop varieties.
组学方法的快速发展对改良经济油料作物的发展起着至关重要的作用。工业油料作物在食品加工、生物燃料、化妆品和制药等行业发挥着至关重要的作用,使它们成为全球经济不可或缺的贡献者,这些作物在众多工业过程中都是至关重要的元素。基因组学的重大进步已经彻底改变了农业部门,特别是在油料作物领域。尖端技术的进步促进了关键商业油料作物基因组的高效测序。这一突破不仅增强了我们对这些作物基因组成的理解,而且还为育种者提供了有针对性的遗传操作和育种计划的宝贵见解。此外,整合转录组学与基因组数据有助于精准农业的新时代。这种方法提供了对相关性状的分子机制的深入了解,如含油量、产量潜力以及对生物和非生物胁迫的抗性。蛋白质组学方法有助于破译蛋白质结构、相互作用和功能的复杂性,而代谢组学和离子学则揭示了生物系统中代谢物和离子的复杂网络。每个组学学科都提供了独特的见解,它们的整合有望丰富我们的理解,并为提高油料作物提供宝贵的见解。本文综述了不同组学方法在工业油料作物精炼中的应用效果和局限性。此外,强调了多组学策略的重要性,并探讨了其与基因工程技术的结合,以培育优质油料作物品种。
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引用次数: 0
Genome-wide identification of the WOX gene family in Populus davidiana×P.bolleana and functional analysis of PdbWOX4 in salt resistance 杨树WOX基因家族的全基因组鉴定davidiana×P。PdbWOX4在抗盐中的功能分析。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-28 DOI: 10.1016/j.plantsci.2024.112379
Zhengyang Li , Ziqian Zhang , Yumeng Xu , Xiaojin Lei , Qinjun Xie , Zhongyuan Liu , Yanmin Wang , Caiqiu Gao
WOX transcription factors (TFs) are plant specific transcription regulatory factors that have a momentous role in maintaining plant growth and development and responding to abiotic stress. In this study, a total of 13 PdbWOX genes were identified. qRT-PCR analyses showed that 13 PdbWOX genes were responsive to salt stress. Notably, the expression of PdbWOX4 was significantly changed at all time points under NaCl stress, suggesting that PdbWOX4 expression may be involved in salt stress. Further, an overexpression vector of PdbWOX4 was constructed and transient transformed into Shanxin poplar. Biochemical staining and physiological parameter analysis showed that overexpression of PdbWOX4 decreased the total antioxidant capacity (T-AOC) and peroxidase (POD) activity, which in turn reduced the scavenging capacity of reactive oxygen species (ROS), and increased the cell damage and death induced by salt stress. qRT-PCR and ChIP-PCR demonstrated that PdbWOX4 can regulate the expression of PdbDREB2C by binding to its promoter. Further analyses revealed that overexpression of PdbDREB2C can reduce cellular damage by increasing ROS scavenging capacity thereby improving salt tolerance in Shanxin poplar. Taken together, we found that PdbWOX4 negatively regulated the salt tolerance of Shanxin poplar by repressing the PdbDREB2C, suggesting that PdbWOX4 may play a key role in the tolerance of Shanxin poplar to salt stress, and is an important candidate gene for molecular resistance breeding in forest trees.
WOX转录因子是植物特异性转录调控因子,在维持植物生长发育和应对非生物胁迫中起着重要作用。本研究共鉴定出13个PdbWOX基因。qRT-PCR分析显示,13个PdbWOX基因对盐胁迫有响应。值得注意的是,在NaCl胁迫下,PdbWOX4的表达在各个时间点都发生了显著变化,表明PdbWOX4的表达可能与盐胁迫有关。构建了PdbWOX4过表达载体,并将其瞬时转化到山西杨树中。生化染色和生理参数分析表明,PdbWOX4过表达降低了细胞的总抗氧化能力(T-AOC)和过氧化物酶(POD)活性,从而降低活性氧(ROS)的清除能力,增加了盐胁迫引起的细胞损伤和死亡。qRT-PCR和ChIP-PCR证实PdbWOX4可以通过结合PdbDREB2C的启动子调控PdbDREB2C的表达。进一步分析发现,过表达PdbDREB2C可以通过增加活性氧清除能力来减轻细胞损伤,从而提高山新杨树的耐盐性。综上所述,我们发现PdbWOX4通过抑制PdbDREB2C负向调控山新杨耐盐性,提示PdbWOX4可能在山新杨耐盐胁迫中起关键作用,是林木分子抗性育种的重要候选基因。
{"title":"Genome-wide identification of the WOX gene family in Populus davidiana×P.bolleana and functional analysis of PdbWOX4 in salt resistance","authors":"Zhengyang Li ,&nbsp;Ziqian Zhang ,&nbsp;Yumeng Xu ,&nbsp;Xiaojin Lei ,&nbsp;Qinjun Xie ,&nbsp;Zhongyuan Liu ,&nbsp;Yanmin Wang ,&nbsp;Caiqiu Gao","doi":"10.1016/j.plantsci.2024.112379","DOIUrl":"10.1016/j.plantsci.2024.112379","url":null,"abstract":"<div><div>WOX transcription factors (TFs) are plant specific transcription regulatory factors that have a momentous role in maintaining plant growth and development and responding to abiotic stress. In this study, a total of 13 <em>PdbWOX</em> genes were identified. qRT-PCR analyses showed that 13 <em>PdbWOX</em> genes were responsive to salt stress. Notably, the expression of <em>PdbWOX4</em> was significantly changed at all time points under NaCl stress, suggesting that <em>PdbWOX4</em> expression may be involved in salt stress. Further, an overexpression vector of <em>PdbWOX4</em> was constructed and transient transformed into Shanxin poplar. Biochemical staining and physiological parameter analysis showed that overexpression of <em>PdbWOX4</em> decreased the total antioxidant capacity (T-AOC) and peroxidase (POD) activity, which in turn reduced the scavenging capacity of reactive oxygen species (ROS), and increased the cell damage and death induced by salt stress. qRT-PCR and ChIP-PCR demonstrated that PdbWOX4 can regulate the expression of <em>PdbDREB2C</em> by binding to its promoter. Further analyses revealed that overexpression of <em>PdbDREB2C</em> can reduce cellular damage by increasing ROS scavenging capacity thereby improving salt tolerance in Shanxin poplar. Taken together, we found that <em>PdbWOX4</em> negatively regulated the salt tolerance of Shanxin poplar by repressing the <em>PdbDREB2C</em>, suggesting that <em>PdbWOX4</em> may play a key role in the tolerance of Shanxin poplar to salt stress, and is an important candidate gene for molecular resistance breeding in forest trees.</div></div>","PeriodicalId":20273,"journal":{"name":"Plant Science","volume":"352 ","pages":"Article 112379"},"PeriodicalIF":4.2,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142907543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 study of the R2R3-MYB gene family and analysis of HhMYB111r-induced salt tolerance in Hibiscus hamabo Sieb. et Zucc 木芙蓉R2R3-MYB基因家族的全基因组研究及hhmyb111r诱导的耐盐性分析调查。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-27 DOI: 10.1016/j.plantsci.2024.112378
Yu Xu , Longjie Ni , Chaoguang Yu , Jianfeng Hua , Yunlong Yin , Chunsun Gu , Zhiquan Wang
Hibiscus hamabo Sieb. et Zucc. (H. hamabo) is a semi-mangrove plant with excellent stress tolerance that plays a crucial role in the ecological restoration of saline and alkaline areas. It is an ideal candidate species for studying the mechanisms involved in stress tolerance. Although the MYB gene family has preliminarily been characterized in H. hamabo, the specific functions and action mechanisms of the R2R3-MYB genes in this species have not fully been elucidated. In this study, 190 R2R3-MYB genes were identified at the genomic level using bioinformatics methods. The genes were divided into 26 subgroups based on their evolutionary relationships and found to be distributed randomly on 46 chromosomes. RNA sequencing data and subsequent real-time quantitative PCR analysis of 12 differentially expressed R2R3-HhMYB genes showed HhMYB111r to be highly expressed under various abiotic stress conditions. Self-activation and subcellular localization results showed that the intact HhMYB111r had strong self-activation activity and located in both the nucleus and cytoplasm. Overexpression in Arabidopsis significantly improved salt tolerance, and silencing HhMYB111r reduced the tolerance of H. hamabo to salt stress, indicating that HhMYB111r positively regulates the salt stress response. In this first analysis of the R2R3-MYB gene family in H. hamabo, we identified a key salt stress response gene, HhMYB111r, enriching the understanding of MYB function and laying a foundation for exploring the abiotic stress response of plants.
芙蓉花。调查。hamabo是一种具有优良抗逆性的半红树林植物,在盐碱地的生态恢复中起着至关重要的作用。它是研究抗逆性机制的理想候选物种。虽然MYB基因家族在hamabo中已被初步鉴定,但R2R3-MYB基因在该物种中的具体功能和作用机制尚未完全阐明。本研究利用生物信息学方法在基因组水平上鉴定了190个R2R3-MYB基因。这些基因根据它们的进化关系被分成26个亚组,并被发现随机分布在46条染色体上。对12个差异表达的R2R3-HhMYB基因的RNA测序数据和实时定量PCR分析显示,在各种非生物胁迫条件下,HhMYB111r均有高表达。自激活和亚细胞定位结果表明,完整的HhMYB111r具有较强的自激活活性,位于细胞核和细胞质中。在拟南芥中过表达HhMYB111r显著提高了盐胁迫的耐受性,而沉默HhMYB111r则降低了H. hamabo对盐胁迫的耐受性,表明HhMYB111r正调控盐胁迫反应。在首次对hamabo R2R3-MYB基因家族的分析中,我们发现了一个关键的盐胁迫应答基因HhMYB111r,丰富了对MYB功能的认识,为探索植物的非生物胁迫应答奠定了基础。
{"title":"Genome-wide study of the R2R3-MYB gene family and analysis of HhMYB111r-induced salt tolerance in Hibiscus hamabo Sieb. et Zucc","authors":"Yu Xu ,&nbsp;Longjie Ni ,&nbsp;Chaoguang Yu ,&nbsp;Jianfeng Hua ,&nbsp;Yunlong Yin ,&nbsp;Chunsun Gu ,&nbsp;Zhiquan Wang","doi":"10.1016/j.plantsci.2024.112378","DOIUrl":"10.1016/j.plantsci.2024.112378","url":null,"abstract":"<div><div><em>Hibiscus hamabo</em> Sieb. et Zucc. (<em>H. hamabo</em>) is a semi-mangrove plant with excellent stress tolerance that plays a crucial role in the ecological restoration of saline and alkaline areas. It is an ideal candidate species for studying the mechanisms involved in stress tolerance. Although the MYB gene family has preliminarily been characterized in <em>H. hamabo</em>, the specific functions and action mechanisms of the R2R3-MYB genes in this species have not fully been elucidated. In this study, 190 R2R3-MYB genes were identified at the genomic level using bioinformatics methods. The genes were divided into 26 subgroups based on their evolutionary relationships and found to be distributed randomly on 46 chromosomes. RNA sequencing data and subsequent real-time quantitative PCR analysis of 12 differentially expressed R2R3-HhMYB genes showed <em>HhMYB111r</em> to be highly expressed under various abiotic stress conditions. Self-activation and subcellular localization results showed that the intact HhMYB111r had strong self-activation activity and located in both the nucleus and cytoplasm. Overexpression in <em>Arabidopsis</em> significantly improved salt tolerance, and silencing <em>HhMYB111r</em> reduced the tolerance of <em>H. hamabo</em> to salt stress, indicating that <em>HhMYB111r</em> positively regulates the salt stress response. In this first analysis of the R2R3-MYB gene family in <em>H. hamabo</em>, we identified a key salt stress response gene, <em>HhMYB111r</em>, enriching the understanding of MYB function and laying a foundation for exploring the abiotic stress response of plants.</div></div>","PeriodicalId":20273,"journal":{"name":"Plant Science","volume":"352 ","pages":"Article 112378"},"PeriodicalIF":4.2,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142903501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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
A new method for identifying proteins involved in DNA methylation through reverse genetics in Arabidopsis 通过反向遗传学鉴定拟南芥DNA甲基化相关蛋白的新方法。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-27 DOI: 10.1016/j.plantsci.2024.112376
Wei Miao , Jie Dai , Li Zhang , Zhile Liang , Xiaoxuan Sun , Meizi Huang , Aqin Zhang , Long Zheng , Yongjun Li , Ying Li
Forward genetic screens have uncovered numerous genes involved in DNA methylation regulation, but these methods are often time-intensive, costly, and labor-intensive. To address these limitations, this study utilized CRISPR technology to knockout selected co-expressed genes, enabling the rapid identification of low luciferase (LUC) luminescence mutants in the Col-LUC line, which harbors a LUC transgene driven by a 2 × 35S promoter in Arabidopsis. As proof of concept, the repressor of silencing 1 (ROS1) and RNA-directed DNA methylation 1 (RDM1) genes were used as controls, while the increased DNA methylation 3 (IDM3) gene, co-expressed with ROS1, was selected as the target for gene knockout experiments. The results demonstrated that combining co-expression analysis with CRISPR technology is an effective strategy for generating low LUC luminescence mutants in the Col-LUC line. Notably, a new mutant, named reduced luminescence 1 (rl1), was identified through this approach. The rl1 mutant exhibited genome-wide DNA hypermethylation, and its reduced luminescence phenotype was largely reversed by treatment with the DNA methylation inhibitor 5-Aza-2’-deoxycytidine, confirming its anti-silencing role in DNA methylation regulation. This study presents a novel and efficient approach for obtaining low luminescence mutants in the Col-LUC line and identifies RL1 as a previously uncharacterized protein involved in DNA methylation regulation.
前向基因筛选已经发现了许多参与DNA甲基化调控的基因,但这些方法通常是耗时、昂贵和劳动密集型的。为了解决这些局限性,本研究利用CRISPR技术敲除选定的共表达基因,从而能够快速鉴定出coll -LUC系中的低荧光素酶(LUC)发光突变体,该系在拟南芥中含有一个由2×35S启动子驱动的LUC转基因。为了证明这一概念,我们以沉默抑制因子1 (ROS1)和rna定向DNA甲基化1 (RDM1)基因作为对照,而选择与ROS1共表达的DNA甲基化3 (IDM3)基因作为基因敲除实验的靶点。结果表明,将共表达分析与CRISPR技术相结合是在Col-LUC系中产生低LUC发光突变体的有效策略。值得注意的是,通过这种方法发现了一个新的突变体,命名为发光减少1 (rl1)。rl1突变体表现出全基因组DNA超甲基化,并且通过DNA甲基化抑制剂5-Aza-2'-脱氧胞苷处理,其减少的发光表型在很大程度上被逆转,证实了其在DNA甲基化调控中的抗沉默作用。本研究提出了一种新的、有效的方法来获得coll - luc系的低发光突变体,并鉴定出RL1是一种以前未被表征的参与DNA甲基化调控的蛋白质。
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引用次数: 0
Involvement of nitrate reductase in nitric oxide generation and in the induction of acclimation responses to phosphorus restriction in soybean plants 硝酸还原酶在大豆氮氧化物生成和对磷限制的驯化反应中的作用。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-26 DOI: 10.1016/j.plantsci.2024.112377
Melisa Luquet , Cristian Antonelli , Gustavo E. Gergoff Grozeff , Agustina Buet , Andrea Galatro
Nitrate reductase (NR) is an essential enzyme because of its role in nitrogen metabolism and in key signaling events through the generation of the reactive nitrogen species, nitric oxide (NO). In this work, we evaluated changes in endogenous NO levels during the onset of P-restriction in soybean plants (Glycine max), focusing on the possible pathways involved in its generation, namely NR and NO synthase like activity, NOSlike, and the subsequent role of NR during low P-acclimation. During the first 96 h of P-starvation NO levels increased in the leaves. The treatment of plants with L-NAME (a known inhibitor of NOS) did not markedly affect NO levels, while the inhibition of NR activity with sodium tungstate (W) reduced NO-related DAF-FM fluorescence. In addition, P-restriction triggered typical acclimation responses such as the increase of acid phosphatase (AP) and ribonuclease (RNase) activities, and the accumulation of organic acids (malic and citric). Treatment of P starved plants with W reduced NR activity as well as the magnitude of the acclimation responses studied. Overall, the data here presented suggest a role for NR as NO source, influencing the signaling pathways leading to early acclimation responses following P-deprivation in soybean plants.
硝酸还原酶(NR)是一种重要的酶,因为它在氮代谢和通过生成活性氮物种一氧化氮(NO)的关键信号事件中发挥作用。在这项工作中,我们评估了大豆植物(Glycine max)在p限制开始时内源NO水平的变化,重点关注其产生的可能途径,即NR和NO合成酶样活性,nos样,以及NR在低p驯化中的后续作用。在磷饥饿的前96h,叶片中NO水平升高。L-NAME(一种已知的NOS抑制剂)对植物的NO水平没有显著影响,而钨酸钠(W)对NR活性的抑制降低了NO相关的DAF-FM荧光。此外,磷限制引发了典型的驯化反应,如酸性磷酸酶(AP)和核糖核酸酶(RNase)活性的增加,有机酸(苹果酸和柠檬酸)的积累。缺磷植物W处理降低了NR活性和驯化响应的幅度。综上所述,本研究的数据表明NR作为NO源,影响了大豆植株在缺磷后早期驯化反应的信号通路。
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引用次数: 0
Genome-wide identification and analysis of autophagy-related (ATG) genes in Lycium ruthenicum Murray reveals their crucial roles in salt stress tolerance 枸杞自噬相关基因(ATGs)的全基因组鉴定和分析揭示了它们在盐胁迫耐受中的重要作用。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-25 DOI: 10.1016/j.plantsci.2024.112371
Yi Shi , Yan Wu , Mengling Li , Na Luo , Faqiang Li , Shaohua Zeng , Ying Wang , Chao Yang
Autophagy is a highly conserved intracellular degradation system that is crucial for nutrient recycling, thus regulating plant growth and development as well as in response to various stresses. Halophytic plant Lycium ruthenicum Murray (L. ruthenicum) is considered as a potential model plant for studying the physiological mechanisms of salt stress tolerance in plants. Although the genome sequence of L. ruthenicum is available, the characteristics and functions of the salt stress-related genes remain largely unknown. In the present study, a total of 36 AuTophaGy-related (ATG) genes were identified in L. ruthenicum and detailed characteristics of them were given. Quantitative real-time polymerase chain reaction analysis revealed that the expression of 25 LrATGs was significantly upregulated after salt stress treatments. Furthermore, the autophagic marker line pSuper:GFP-LrATG8g was generated and used to demonstrate the salt stress-induced autophagy, as revealed by measuring autophagic flux and observing autophagosome formation. The pSuper:LrATG5-GFP overexpression (OE) lines were also generated and further phenotypic analysis showed that OE-LrATG8g and OE-LrATG5 plants exhibited better salt tolerance than that of WT plants. To the best of our knowledge, this study firstly reports a detailed overview of LrATGs-mediated autophagy in L. ruthenicum response to salt stress. These findings contribute to a global understanding of the characteristics of ATG genes in L. ruthenicum and lay a foundation for future functional study.
自噬是一种高度保守的细胞内降解系统,对养分循环至关重要,从而调节植物的生长发育以及对各种胁迫的反应。盐生植物枸杞(L. ruthenicum)被认为是研究植物耐盐生理机制的潜在模式植物。虽然已知盐胁迫相关基因的基因组序列,但对盐胁迫相关基因的特征和功能仍知之甚少。本研究共鉴定了36个自噬相关基因(LrATGs),并给出了它们的详细特征。定量实时聚合酶链反应分析显示,盐胁迫处理后,25个LrATGs的表达显著上调。通过测定自噬通量和观察自噬体的形成,制备了自噬标记系pSuper:GFP-LrATG8g,用于证明盐胁迫诱导的自噬。进一步的表型分析表明,OE- lratg8g和OE- lratg5植株的耐盐性优于WT植株。据我们所知,本研究首次报道了lratgs介导的自噬在L. ruthenicum对盐胁迫的反应中的详细概述。这些发现有助于全面了解白乳草ATG基因的特征,为今后的功能研究奠定基础。
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引用次数: 0
ArHDZ19 contributes to drought tolerance by advancing flowering time in Anoectochilus roxburghii ArHDZ19通过提前开花来提高野樱草的耐旱性。
IF 4.2 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-12-25 DOI: 10.1016/j.plantsci.2024.112369
Yanqin Zhu , Xiaohui Zhou , Jinjing Shi , Bingcong Xing , Ying Zheng , Siqi Wan , Lan ying Pan , Aimin Lv , Qingsong Shao
The homeodomain-leucine zippper gene family encodes plant-specific transcription factors that not only affect growth and development, but also play important roles in the drought response. ArHDZ19, from Anoectochilus roxburghii, encodes a homeodomain-leucine zipper III subfamily protein whose function and molecule ar mechanism remains unclear. Here, we explored the function of ArHDZ19 in regulating growth and the drought response. ArHDZ19 localized in the nucleus and its expression was strongly induced under drought stress. Overexpression of ArHDZ19 in Arabidopsis thaliana (OE-ArHDZ19) not only increased plant height and the length of stamens and pistils, but also resulted in an earlier flowering phenotype. The flowering-related genes FLOWERING LOCUS T (FT), SUPPRESSOR OF CONSTANS OVEREXPRESSION 1 (SOC1), CONSTANS (CO), FLOWERING LOCUS C (FLC), and GIGANTEA (GI) were up-regulated in the OE-ArHDZ19 lines. Moreover, under drought conditions, overexpression of ArHDZ19 improved the plant survival rate and advanced the flowering time. Stress-responsive genes such as COLD-REGULATED 47 (COR47), KINESIN 1 (KIN1), and RESPONSE TO DESICCATION (RD29A) were up-regulated by drought treatment; however, their transcript levels were lower in OE-ArHDZ19 plants than in wild-type plants. These results indicate that ArHDZ19 can improve the drought resistance of plants by advancing the flowering time, which may be a drought-avoidance mechanism.
homedomain- leucinezipper基因家族编码的植物特异性转录因子不仅影响植物的生长发育,而且在干旱响应中发挥重要作用。ArHDZ19编码同源结构域亮氨酸zipper III亚家族蛋白,其功能和分子机制尚不清楚。本研究探讨了ArHDZ19在调节植物生长和干旱响应中的作用。ArHDZ19定位于细胞核,干旱胁迫强烈诱导其表达。ArHDZ19在拟南芥中的过表达(e -ArHDZ19)不仅增加了植株的株高和雄蕊、雌蕊的长度,而且使开花表型提前。OE-ArHDZ19株系开花相关基因开花位点T (FT)、CONSTANS过表达抑制因子1 (SOC1)、CONSTANS (CO)、开花位点C (FLC)和GIGANTEA (GI)上调。此外,在干旱条件下,ArHDZ19过表达提高了植株成活率,并提前了开花时间。胁迫应答基因如COLD-REGULATED 47 (COR47)、KIN1 (KIN1)和RESPONSE TO des干(RD29A)在干旱处理下表达上调;然而,它们在OE-ArHDZ19植物中的转录水平低于野生型植物。上述结果表明,ArHDZ19可通过提前开花时间提高植物抗旱性,这可能是一种抗旱机制。
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引用次数: 0
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Plant Science
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