Horticultural Plant Journal最新文献

{"title":"Evaluating the canopy light environment, photosynthesis, and fruit comprehensive performance of greenhouse tomato under different mechanized planting layouts","authors":"Xiaolong Ma, Jiayue Chang, Xinru Chai, Yan Liu, Jianming Li","doi":"10.1016/j.hpj.2025.03.010","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.03.010","url":null,"abstract":"Plant layout is critical for the formation of canopy microenvironment, and fruit yield and quality. Plant-layout designs that meet the requirements of photosynthetically efficient and mechanizable greenhouse cultivation are urgently needed. Here, we report our findings from a two-year trial using tomatoes grown in a plastic film greenhouse at high planting density (HPD, 4.55 plants · m<ce:sup loc=\"post\">−2</ce:sup>) or low planting density (LPD, 3.13 plants · m<ce:sup loc=\"post\">−2</ce:sup>). Each density comprised south-north (SN) and east-west (EW) row orientations, with three plant layouts (S1–S3 or E1-E3, S and E represent SN and EW row orientations, respectively) in each orientation: 90 cm + 35 cm + 35 cm, 90 cm + 60 cm + 30 cm, and 90 cm + 90 cm + 25 cm for HPD; and 120 cm + 40 cm + 40 cm, 120 cm + 60 cm + 36 cm, and 120 cm + 120 cm + 27 cm for LPD. Analysis included canopy accumulated light absorption (ACL), photosynthesis (ACP), and light uniformity (UC<ce:inf loc=\"post\">LP</ce:inf>) conducted using a three-dimensional canopy photosynthesis model, along with individual fruit indices, and a comprehensive evaluation using the Technique for Order Preference by Similarity to an Ideal Solution (TOPSIS). Plants grown under S2 and E2 exhibited respective dominance in both planting densities compared to those grown under any other treatment. Particularly, S2 showed higher ACL and single fruit weight, whereas E2 showed superior UC<ce:inf loc=\"post\">LP</ce:inf>, comprehensive fruit quality, and fruit uniformity. Therefore, E2 was identified as an optimal plant configuration for both HPD and LPD based on overall fruit performance as evaluated using the TOPSIS method. Correlation analysis indicated that the light environment of the lower canopy layer under HPD and that of the middle canopy layer under LPD showed the strongest correlations with the main fruit indices. Overall, the optimal plant layout identified herein provides valuable decision support for horticultural production under mechanized planting conditions. The correlations between canopy light and fruit indices define quantitative breeding and management targets for optimizing fruit performance through adjustments in plant structure.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"205 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144613340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Telomere-to-Telomere Genome of Selaginella moellendorffii Provides Insights into Genome Evolution and Biflavone Biosynthesis","authors":"Hui Xiong, Cong Yin, Ding Tang, Xiran Xiong, Xinqiao Liu, Zhengwen Wang, Xiaolei Yu, Zhinan Mei, Juan Li","doi":"10.1016/j.hpj.2025.04.007","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.04.007","url":null,"abstract":"<ce:italic>Selaginella moellendorffii</ce:italic> Hieron., a lycophyte of significant medicinal and evolutionary importance, is recognized as one of the earliest vascular plants. However, the absence of a high-quality reference genome has hindered the comprehensive exploration of its unique phylogenetic position and therapeutic potential, thereby limiting our understanding of its genomic structure and metabolic capabilities. In this study, we present the first chromosome-level, telomere-to-telomere (T2T) genome assembly of <ce:italic>S. moellendorffii</ce:italic>, constructed utilizing PacBio HiFi, Oxford Nanopore (ONT), and Hi-C technologies. The assembled genome, spanning 112.83 Mb across 10 chromosomes with a contig N50 of 11.11 Mb, exhibited exceptional completeness (BUSCO score: 95.7 %) and accuracy (QV = 48.11). Comparative genomic analysis identified 3 515 gene families unique to <ce:italic>S. moellendorffii</ce:italic>, with significant enrichment in secondary metabolism pathways, including those related to flavonoid biosynthesis. Phylogenetic analysis revealed that <ce:italic>S. moellendorffii</ce:italic> diverged from <ce:italic>Isoetes</ce:italic> approximately 339.6 million years ago (MYA), representing a key evolutionary transition in early vascular plants. By integrating tissue-specific transcriptome and metabolome analyses, we uncovered the molecular basis of biflavone biosynthesis, identifying key enzymes and regulatory networks that govern the production of these bioactive compounds. We observed a correlation between the tissue-specific accumulation patterns of six major biflavones, including amentoflavone and ginkgetin, and the expression of their biosynthetic genes. This high-quality genome assembly, coupled with multi-omics analyses, offers unprecedented insights into the evolution of early vascular plants and elucidates the molecular mechanisms behind their specialized metabolism.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"11 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144613299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gibberellins and their interplay with other hormones in the regulation of fruit ripening","authors":"Tu Feng, Wenbo Leng, Yingdie Yang, Hong Zhao, Zixin Zhang, Mingchun Liu, Gaofeng Liu","doi":"10.1016/j.hpj.2025.04.006","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.04.006","url":null,"abstract":"Gibberellins (GAs) are a class of tetracyclic diterpenoid phytohormones involved in a wide range of plant growth and developmental processes, including seed germination, stem elongation, flower and fruit development. Fruit ripening is a complex process mediated by a regulatory network consists of plant hormones, key transcription factors and epigenetics. Ethylene plays a key role in climacteric fruit ripening, but the involvement of other phytohormones in ripening process remains largely unexplored. Recently, there are increasing evidence showed that GA also play an important role in fruit ripening and related quality regulation. Here, we summarize the recent advances in GA biosynthesis, signaling transduction and their involvement in fruit ripening. Moreover, we discuss the role of GAs in regulating ripening-related traits including softening, color, and flavor formation, as well as their interactions with other hormones in controlling fruit ripening. By integrating insights into GA-mediated control of cell wall integrity, pigment metabolism, and hormonal crosstalk, this review highlights the potential of manipulating GA pathways to optimize fruit quality and postharvest shelf life.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"37 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms and management strategies for rain-cracking in greenhouse and open-air cherry","authors":"Shuo Yang, Zilong Huang, Fuming Yang, Yujie Chi, Yuan Chi","doi":"10.1016/j.hpj.2025.04.008","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.04.008","url":null,"abstract":"Rain-induced cracking remains a persistent challenge in cherry (<ce:italic>Prunus avium</ce:italic>) cultivation, affecting both greenhouse and open-air grown varieties. This review explores and contrasts the underlying mechanisms of rain-cracking in these two cultivation environments, emphasizing key differences in water transport pathways. While rain-cracking in open-air cherries is predominantly driven by xylem water transport, greenhouse cherries are primarily affected by water absorption through the fruit epidermis. Additional distinctions include environmental influences such as calcium leaching from the pericarp and temperature drops caused by wind in open-air cherry, as well as the challenges associated with overthinning in greenhouse cultivation. To mitigate rain-cracking, this review summarizes various orchard management strategies, including crop load regulation, mineral sprays, bio-stimulants, growth regulators, anti-transpirants, edible coatings, and the air-blast sprayer integrated with field-sensing technology. Each approach is evaluated in terms of environmental suitability, efficacy, technical feasibility, and research advancements at both domestic and international levels. Furthermore, key assessment methodologies, including cracking index (CI), T50/WU50, and cracking ratio (CR), are analyzed and compared to establish a standardized framework for evaluating rain-cracking susceptibility. By providing a comprehensive synthesis of current knowledge, this review offers guidance for cherry orchard management and lays the foundation for future research on effective rain-cracking mitigation strategies across diverse cultivation conditions.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"29 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Salt stress response pathway and regulatory mechanism of the Malus domestica G protein-coupled receptor MdGPCR","authors":"Ziquan Feng, Lin Zhao, Tong Li, Xinyi Li, Shuo Ma, Huaina Gao, Ruxue Sha, Ge Tian, Xinxiang Xu, Yue Xing, Mengxue Lyu, Jingquan Liu, Shunfeng Ge, Zhanling Zhu, Yuanyuan Li, Han Jiang, Yuanmao Jiang","doi":"10.1016/j.hpj.2025.03.009","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.03.009","url":null,"abstract":"China accounts for about half of the world's apple area and production, and is a pillar industry that promotes farmers' income growth and rural revitalization. In recent years, with the dramatic change of the global climate, most apple (<ce:italic>Malus domestica</ce:italic>) producing areas in China have suffered salt damage. Soil salinization, especially secondary salinization, is common in apple producing areas in the North China Plain around the Bohai Sea. These phenomena indicate that the ability of apple industry to prevent and resist soil salinization still needs to be improved. In this article, we identified a novel salt sensitive gene <ce:italic>MdGPCR</ce:italic> in apple. We conducted salt stress experiments on apple callus and leaves using <ce:italic>MdGPCR</ce:italic>. The results showed that after salt stress, overexpression of <ce:italic>MdGPCR</ce:italic> in transgenic apple would accumulate a large amount of reactive oxygen species, leading to an imbalance of redox levels in their bodies. At the same time, MdGPCR interacts with MdSOS3 and promotes its degradation, further reducing its salt stress resistance.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"48 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MdCAX5 plays a critical role in calcium homeostasis and bitter pit development in apple","authors":"Jia Liu, Yingwei Qi, Weijie Yin, Xiwen Li, Caixia Wang, Xiaolin Ren","doi":"10.1016/j.hpj.2025.03.008","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.03.008","url":null,"abstract":"Bitter Pit (BP) is a prevalent physiological disorder in apple that significantly reduces fruit quality and market value. While numerous studies have investigated the mechanisms underlying BP occurrence, the molecular processes, particularly the role of the Ca<ce:sup loc=\"post\">2+</ce:sup>/H<ce:sup loc=\"post\">+</ce:sup> exchanger (CAX), remain unclear. This study aims to elucidate the function of the <ce:italic>MdCAX5</ce:italic> gene in relation to BP development. To achieve this, we utilized transient transformation in apple, as well as stable transformation in <ce:italic>Arabidopsis</ce:italic> and tomato, to measure the mineral content in transgenic plants, thereby validating the function of <ce:italic>MdCAX5</ce:italic>. The overexpression of the <ce:italic>MdCAX5</ce:italic> gene significantly reduced calcium (Ca) content in plants and disrupted the mineral element balance within the plant. Analysis of the <ce:italic>MdCAX5</ce:italic> gene promoter revealed that Ca<ce:sup loc=\"post\">2+</ce:sup> can enhance promoter activity, indicating that the <ce:italic>MdCAX5</ce:italic> gene can effectively respond to Ca signaling. Transcriptomic analysis of tomato plants stably overexpressing the <ce:italic>MdCAX5</ce:italic> gene revealed significant alterations in the expression of genes involved in Ca signal transduction and transport, which in turn impacted the biosynthesis of secondary metabolites and metabolic pathways within the plants. These changes resulted in a reduction in Ca content, imbalanced Ca distribution, increased hydrolase activity, and disrupted cellular structures, including compromised organelles, cellular membranes, and membrane components. These disruptions culminated in the manifestation of Ca deficiency symptoms in the plants. This study provides theoretical insights into the mechanisms underlying the occurrence of apple BP disease.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"46 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515924","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-density genetic mapping and QTL analysis for key horticultural traits in bitter gourd : Insights into plant architecture, fruit development, and wart characteristics","authors":"Feifan Chen, Jiayu Liu, Hongrong Chen, Yi Yang, Xuzhen Li, Yuehong Ding, Libo Tian, Sang Shang","doi":"10.1016/j.hpj.2025.03.007","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.03.007","url":null,"abstract":"Bitter gourd (<ce:italic>Momordica charantia</ce:italic> L.), a staple crop in subtropical and tropical Asia and Africa, is valued for its nutritional and medicinal properties. Despite its significance, genetic studies on key horticultural traits remain limited. To address this, we generated F<ce:inf loc=\"post\">1</ce:inf>, F<ce:inf loc=\"post\">2</ce:inf>, and F<ce:inf loc=\"post\">2:3</ce:inf> populations by crossing two inbred lines, HNU004 and HNU025. Whole-genome sequencing of the parents and 178 F<ce:inf loc=\"post\">2</ce:inf> individuals enabled the construction of a high-density genetic map with 2,605 bin SNP markers across 11 linkage groups. QTL mapping using Composite Interval Mapping (CIM) and Multiple QTL Model (MQM) approaches across three environments identified 22 QTLs influencing plant architecture, fruit size, and wart traits, explaining 6.14–68.12% of phenotypic variance. This included three novel QTLs for average internode length, three for lateral branch number (LBN), and four for fruit-related traits. Five key QTLs were consistently detected: <ce:italic>lbn3.1</ce:italic> on chromosome 3 (LBN), <ce:italic>mfl5.1</ce:italic> and <ce:italic>smfw5.1</ce:italic> on chromosome 5 (mature fruit length and single mature fruit weight), and <ce:italic>fwf4.1</ce:italic> and <ce:italic>fwf6.1</ce:italic> on chromosomes 4 and 6 respectively governing fruit wart characteristics. These QTLs spanned physical regions ranging from 220 kb to 2.1 Mb. Candidate genes were predicted for major QTLs, including <ce:italic>Moc03g28260</ce:italic> (<ce:italic>lbn3.1</ce:italic>), <ce:italic>Moc05g28880</ce:italic> and <ce:italic>Moc05g29850</ce:italic> (<ce:italic>mfl5.1</ce:italic>), and <ce:italic>Moc06g04450</ce:italic> and <ce:italic>Moc06g04970</ce:italic> (<ce:italic>fwf6.1</ce:italic>). Epistatic interactions between QTLs for LBN and fruit length suggested complex genetic regulation. Molecular markers for <ce:italic>mfl5.1</ce:italic>, <ce:italic>fwf4.1</ce:italic>, and <ce:italic>fwf6.1</ce:italic> were validated in an independent F<ce:inf loc=\"post\">2</ce:inf> population of 213 individuals, which confirmed their phenotypic effects. This study provides a dense and informative set of genetic markers suitable for marker-assisted selection in bitter gourd breeding and establishes a foundation for the cloning of candidate genes, thereby accelerating genetic improvement efforts.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"149 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of plant factory with artificial lighting in horticultural production: current progress and future trends","authors":"Meng Guo, Zhi Jin, Li Ma, Siying Ou","doi":"10.1016/j.hpj.2025.04.005","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.04.005","url":null,"abstract":"Horticultural production, enriched in essential nutrients such as proteins, carbohydrates, vitamins, and lipids, face challenges due to rapid global population growth, resource shortages, environmental degradation, and a decline in the number of horticultural practitioners. These challenges prevent their yield and quality from meeting sustainability requirements. By regulating temperature, humidity, light, and nutrient supply, plant factories with artificial lighting (PFALs) enable stable crop cultivation, facilitate year-round production, and promote efficient resource use, yielding high-quality horticultural products with significant application potential. However, issues such as high initial investment and electricity costs remain unresolved. Primarily, PFALs are utilized in the production of green leafy vegetables, transplants, and medicinal plants. In PFALs, lettuce yield can increase by over 50% under optimal lighting conditions compared to traditional methods. PFALs are also employed in the commercial production of horticultural seedlings, technical research on shortening growth cycle and mechanism research on environmental regulation of metabolites. This supports rapid breeding and the production of high-value horticultural products. Additionally, this paper addresses the challenges faced by plant factories in horticultural production and explores prospects and considerations for PFAL applications in this domain.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"30 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of genes involved in the tomato Mi-1-mediated immunity to root-knot nematodes parasitism under high temperature","authors":"Yuqing Lai, Zhijie Li, Erfeng Li, Rui Liu, Nv Chang, Cuihua Lu, Jian Ling, Xiaoxiao Zhang, Shanshan Yang, Zhukan Chen, Yuhong Yang, Yan Li, Bingyan Xie, Zhenchuan Mao, Jianlong Zhao","doi":"10.1016/j.hpj.2025.02.021","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.02.021","url":null,"abstract":"Root-knot nematodes (RKNs) are notorious plant pathogens that cause substantial losses in horticultural crops globally. The tomato RKN resistance gene, <ce:italic>Mi-1</ce:italic>, is currently the sole commercially available source of resistance. However, its effectiveness significantly declines when soil temperatures exceed 28 °C. Although numerous studies have explored the immune signaling pathways mediated by <ce:italic>Mi-1</ce:italic>, the specific plant immune responses affected by high temperatures and the key temperature-sensitive genes involved remain poorly understood. In this study, we demonstrated that the <ce:italic>Mi-1</ce:italic>-mediated hypersensitive response is impaired at 32 °C. At this temperature, the production of reactive oxygen species (ROS) in roots is reduced, while callose deposition increases. Through comparative transcriptome analysis between 24 °C and 32 °C, we identified significant changes in hormone signaling pathways, immune signaling, and gene alternative splicing. For instance, the jasmonate (JA) pathway was upregulated, and the salicylic acid (SA) pathway was inhibited at 32 °C. High temperatures also disrupted the MAPK cascade and influenced metabolite synthesis. Notably, most genes upregulated at 24 °C were downregulated at 32 °C. Furthermore, virus-induced gene silencing (VIGS) assays verified that interfering with the expression of differentially expressed genes, such as the JA biosynthesis key gene MYB transcription factor <ce:italic>AOS3</ce:italic>, the abscisic acid (ABA) synthesis regulation gene <ce:italic>JA2</ce:italic>, and the heat stress transcription factor <ce:italic>A-6b</ce:italic>, increased the susceptibility of <ce:italic>Mi-1</ce:italic> tomatoes to RKNs. These findings offer crucial insights into the temperature sensitivity of <ce:italic>Mi-1</ce:italic> resistance and support the development of RKN-resistant tomatoes that can remain effective under high-temperature conditions.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"47 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515925","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and activity evaluation of a highly efficient CRISPR-Cas genome editing system in larch","authors":"Xuelian Zheng, Lei Ding, Guanqin Liu, Jiong Guo, Zhe Zhang, Jingru Zhang, Yunjin Pang, Xu Tang, Qiurong Ren, Binglin Liu, Lan Huang, Tao Zhang, Yong Zhang","doi":"10.1016/j.hpj.2025.02.020","DOIUrl":"https://doi.org/10.1016/j.hpj.2025.02.020","url":null,"abstract":"The CRISPR/Cas system has been widely used among plant species. However, due to the long life cycle, complex genetic background, high levels of genomic heterozygosity, and difficulties in stable transformation, genome editing has not been thoroughly explored in larch (<ce:italic>Larix kaempferi</ce:italic> (Lamb.) Carr.). In this study, we evaluated a highly efficient CRISPR/Cas genome editing system in larch that utilizes endogenous promoters. We optimized the critical parameters in larch protoplast preparation and transformation. As a result, this process achieved more than 90% active cells and a transient transformation efficiency of 40%, meeting the requirements for gene editing system evaluation. Using integrated whole-genome and transcriptome sequencing analysis, we screened highly expressed mRNA transcripts and cloned 41 candidate promoters. Based on protoplast transient expression detection, we identified a highly expressed endogenous promoter in larch, designated LarPE004. We compared the single and multiple gene knockout capabilities of the LarPE004-driven single transcription unit CRISPR-Cas9 (STU-Cas9) and two transcription unit CRISPR-Cas9 (TTU-Cas9) systems. Our findings showed that the LarPE004::STU-Cas9 system was more efficient than the LarPE004::TTU-Cas9 system and significantly outperformed the CaMV 35S- and ZmUbi1-driven STU-Cas9 systems. Moreover, the LarPE004::STU-Cas9 system demonstrated improved capabilities for multiple gene editing in larch. The LarPE004::STU-SpRY system based on Cas9 mutant protein SpRY facilitated the efficient editing of various PAM sites in larch. Combining these findings, we developed LarPE004::STU-Cas9, an efficient CRISPR-Cas9 genome system for single and multiple gene editing, establishing a foundation for functional genomics research and molecular breeding in larch.","PeriodicalId":13178,"journal":{"name":"Horticultural Plant Journal","volume":"244 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}