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Rapid isolation of Yr9 via MutIsoSeq and QTL analysis of durable stripe rust resistance in wheat cultivar Xingzi 9104. 利用mutisseq快速分离小麦品种Yr9并进行星子9104抗持久条锈病的QTL分析。
Pub Date : 2025-04-30 DOI: 10.1007/s44154-025-00226-9
Yibo Zhang, Shuo Huang, Yuqing Li, Shuaiwei Cao, Hui Ren, Mingjie Xiang, Haitao Dong, Jiangna Han, Ying Zhao, Xiangxue Zhang, Xunying Yuan, Qilin Wang, Yajun Wang, Yi Ouyang, Zujun Yang, Zhensheng Kang, Shengjie Liu, Jianhui Wu, Qingdong Zeng, Dejun Han

The fungus Puccinia striiformis f. sp. tritici (Pst) is the causal agent of wheat stripe rust which constitutes a major limitation to wheat production. Cloning and applying disease-resistant genes are considered as an effective solution. Chinese wheat cultivar Xingzi 9104 (XZ9104) has exhibited durable resistance across multiple environments since its release. Through quantitative trait loci (QTL) analysis, eight QTL were found on chromosome arms 1BS, 1BL, 2AL, 2BL, 3BS, 4BL, 5BL and 7BL. YrXZ identified as 1RS.1BL translocation conferred race-specific all-stage resistance to Pst race CYR23. QYrxz.nwafu-1BL.6 and QYrxz.nwafu-3BS.7 were considered as the adult plant resistance genes Yr29 and Yr30, respectively. Notably, QYrxz.nwafu-2BL.5 accounted for 15.75-47.63% of the phenotypic variation across diverse environments and its pyramiding with Yr29 and Yr30 can confer high level of resistance. Other QTL were environment-dependent with minor effects. To clone the above resistance genes, we created a population of over 2,000 M5 mutants in XZ9104 using ethylmethane sulfonate (EMS) mutagenesis and screened various types of susceptible mutants. Using the MutIsoseq approach with five mutant lines susceptible to race CYR23, we rapid isolated a candidate gene for YrXZ encoding coiled-coil nucleotide-binding site leucine-rich repeat (CC-NBS-LRR) protein. Integrating cytological analysis, gene-based association analysis, transcriptomic profiling and virus-induced gene silencing (VIGS), we confirmed that the causal gene for YrXZ was indeed Yr9. This study demonstrated that multiple QTL with different effects contributed to the durable resistance in XZ9104. Understanding the molecular mechanisms and pathways involved in plant defense can inform future strategies for deploying resistance gene and engineering of genetic resistance against evolving diseases.

小麦条锈病是小麦条锈病的主要病原菌,是制约小麦生产的主要病原菌。克隆和应用抗病基因被认为是有效的解决办法。我国小麦品种星子9104 (XZ9104)自投放以来,在多种环境中表现出持久的抗性。通过数量性状位点(QTL)分析,在染色体臂1BS、1BL、2AL、2BL、3BS、4BL、5BL和7BL上发现8个QTL。YrXZ被识别为1RS。1BL易位赋予了对Pst小种CYR23的全阶段特异性抗性。QYrxz.nwafu-1BL。6和QYrxz.nwafu-3BS。其中,Yr29和Yr30分别为成虫抗性基因。值得注意的是,QYrxz.nwafu-2BL。5在不同环境下占表型变异的15.75-47.63%,与Yr29和Yr30的金字塔化可以获得高水平的抗性。其他QTL与环境相关,影响较小。为了克隆上述抗性基因,我们在XZ9104上利用甲烷磺酸乙酯(EMS)诱变技术建立了2000多个M5突变体群体,并筛选了不同类型的易感突变体。利用对CYR23易感的5个突变系的mutisseq方法,我们快速分离出了YrXZ编码卷曲卷曲核苷酸结合位点富亮氨酸重复序列(CC-NBS-LRR)蛋白的候选基因。综合细胞学分析、基因关联分析、转录组学分析和病毒诱导基因沉默(VIGS),我们证实YrXZ的致病基因确实是Yr9。本研究表明,XZ9104的持久抗性是由多个不同作用的QTL促成的。了解植物防御的分子机制和途径可以为未来抗病基因的部署和抗病基因工程提供信息。
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引用次数: 0
Plant-aphid interactions: recent trends in plant resistance to aphids. 植物与蚜虫的相互作用:植物抗蚜虫的最新趋势。
Pub Date : 2025-04-29 DOI: 10.1007/s44154-025-00214-z
Kifle Gebreegziabiher Gebretsadik, Zhixin Liu, Jincheng Yang, Hao Liu, Aizhi Qin, Yaping Zhou, Enzhi Guo, Xiao Song, Peibo Gao, Yajie Xie, Ninkuu Vincent, Lam-Son Phan Tran, Xuwu Sun

Aphids are highly destructive agricultural pests characterized by complex life cycles and phenotypic variability, facilitating their adaptation to diverse climates and host plants. Their feeding behavior leads to plant deformation, wilting, stunted growth, disease transmission, and significant yield losses. Given the economic risks aphids pose, regular updates on their seasonal behaviors, adaptive mechanisms, and destructive activities are critical for improving management strategies to mitigate crop losses. This review comprehensively synthesizes recent studies on aphids as plant pests, the extrinsic factors influencing their life cycles, and the intricate interactions between aphids and their hosts. It also highlights recent advancements in biological control measures, including natural enemies, antibiosis, and antixenosis. Additionally, we explore plant defense mechanisms against aphids, focusing on the roles of cell wall components such as lignin, pectin and callose deposition and the genetic regulations underlying these defenses. Aphids, however, can evolve specialized strategies to overcome general plant defenses, prompting the development of targeted mechanisms in plants, such as the use of resistance (R) genes against specific aphid species. Additionally, plant pattern recognition receptors (PRRs) recognize compounds in aphid saliva, which triggers enhanced phloem sealing and more focused immune responses. This work enhances understanding of aphid-plant interaction and plant resistance and identifies key research gaps for future studies.

蚜虫是具有高度破坏性的农业害虫,其特征是复杂的生命周期和表型变异性,有助于它们适应不同的气候和寄主植物。它们的取食行为导致植物变形、枯萎、生长发育迟缓、疾病传播和显著的产量损失。鉴于蚜虫带来的经济风险,定期更新蚜虫的季节性行为、适应机制和破坏性活动对于改善管理策略以减轻作物损失至关重要。本文综述了近年来蚜虫作为植物害虫的研究进展、影响蚜虫生命周期的外来因素以及蚜虫与寄主之间复杂的相互作用。它还强调了生物防治措施的最新进展,包括天敌、抗生素和抗虫病。此外,我们还探讨了植物对蚜虫的防御机制,重点研究了木质素、果胶和胼胝质沉积等细胞壁成分的作用以及这些防御的遗传调控。然而,蚜虫可以进化出特殊的策略来克服一般的植物防御,促进植物中靶向机制的发展,例如使用抗性(R)基因来对抗特定的蚜虫物种。此外,植物模式识别受体(PRRs)识别蚜虫唾液中的化合物,从而增强韧皮部密封和更集中的免疫反应。这项工作提高了对蚜虫与植物相互作用和植物抗性的理解,并为未来的研究确定了关键的研究空白。
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引用次数: 0
Development of novel InDel markers by whole-genome sequence comparison and genetic diversity assessment of Thailand rice blast fungus populations. 利用泰国稻瘟病菌群体全基因组序列比较和遗传多样性评估开发新的InDel标记。
Pub Date : 2025-04-27 DOI: 10.1007/s44154-025-00212-1
Napassorn Thamkirati, Worrawit Suktrakul, Athipat Ngernmuen, Theerayut Toojinda, Sureeporn Katengam, Nonglak Parinthawong, Waree Laophermsuk, Pradipha Pradapphai, Watchareeporn Suksiri, Suphattra Janthasri, Chatchawan Jantasuriyarat

InDel markers are commonly used to assess genetic relationships among populations. In this study, we employed a whole-genome sequence comparison method to identify and develop InDel markers for the rice blast fungus Pyricularia oryzae. We analyzed 152 whole-genome sequences of P. oryzae isolates from diverse global regions, including Brazil, Burundi, China, Colombia, Côte d'Ivoire, France, Ghana, Hungary, India, Japan, Korea, Laos, Madagascar, Mali, Morocco, Nepal, the Philippines, Portugal, Spain, Suriname, Thailand, the UK, the USA, and Zambia. Our analysis identified a total of 233,595 InDel loci distributed across the seven chromosomes of P. oryzae. From these, 82 loci were selected based on their high polymorphism across the 152 genome sequences. The effectiveness of these 82 loci was assessed by analyzing the genetic diversity of 47 Thai rice blast isolates alongside two reference isolates, GUY11 (France) and KJ201 (Korea). Of the 82 InDel loci, 33 exhibited polymorphisms, with 2-4 alleles per locus and polymorphic information content (PIC) scores ranging from 0.04 to 0.67. Principal coordinate and structure analyses revealed two genetic subgroups among the Thai rice blast isolates, categorized according to host specificity. Genetic relationships highlighted disparities among rice blast populations based on their respective hosts: rice and grassy weeds. This finding suggests a correlation between genetic relatedness and the plant hosts susceptible to rice blast disease. The newly developed InDel markers provide a valuable resource for future research in this field.

InDel标记通常用于评估群体间的遗传关系。本研究采用全基因组序列比对的方法,对稻瘟病菌稻瘟病菌(pyricaria oryzae)进行InDel标记鉴定和开发。我们分析了来自巴西、布隆迪、中国、哥伦比亚、Côte科特迪瓦、法国、加纳、匈牙利、印度、日本、韩国、老挝、马达加斯加、马里、摩洛哥、尼泊尔、菲律宾、葡萄牙、西班牙、苏里南、泰国、英国、美国和赞比亚等全球不同地区的P. oryzae分离株的152个全基因组序列。我们的分析共鉴定出233,595个InDel位点分布在P. oryzae的7条染色体上。从这些基因座中,根据它们在152个基因组序列中的高多态性选择了82个位点。通过分析47个泰国稻瘟病菌株和2个参考菌株GUY11(法国)和KJ201(韩国)的遗传多样性,评价了这82个位点的有效性。在82个InDel位点中,33个位点表现出多态性,每个位点有2-4个等位基因,多态性信息含量(PIC)得分在0.04 ~ 0.67之间。主坐标和结构分析揭示了泰国稻瘟病菌株的两个遗传亚群,根据宿主特异性进行分类。遗传关系突出了稻瘟病种群之间基于各自寄主(水稻和草杂草)的差异。这一发现表明遗传亲缘关系与易患稻瘟病的植物宿主之间存在相关性。新开发的InDel标记为今后该领域的研究提供了宝贵的资源。
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引用次数: 0
The resistance risk of fluopicolide and resistance-associated point mutations in the target protein PlVHA-a in Phytophthora litchii. 荔枝疫霉氟霉酸酯的抗性风险及靶蛋白PlVHA-a的抗性相关点突变
Pub Date : 2025-04-21 DOI: 10.1007/s44154-025-00218-9
Tan Dai, Jikun Yang, Shiping Hu, Chuang Zhao, Kang Yuan, Jianqiang Miao, Xili Liu

Litchi, a fruit that is highly sought-after worldwide, faces significant yield challenges due to litchi downy blight, primarily caused by Phytophthora litchii. Fluopicolide has exhibited remarkable efficacy in inhibiting this pathogen and is utilized for the management of litchi downy blight. Although understanding the resistance of P. litchii to fluopicolide is critical, studies on its risk and mechanisms remain limited. In this study, we determined the sensitivity of 125 P. litchii isolates to fluopicolide, revealing an average EC50 value of 0.131 ± 0.037 μg/mL. Through fungicide adaptation, four resistant mutants were obtained with resistance factors exceeding 600, indicating that these strains exhibited high levels of resistance. A compound fitness index analysis demonstrated that the survival fitness of resistant mutants was significantly lower than that of their parental strains. Cross-resistance assays revealed no cross-resistance between fluopicolide and other fungicides with different modes of action. However, positive cross-resistance was observed with fluopimomide. A comprehensive evaluation suggested a moderate risk of P. litchii developing resistance to fluopicolide. PlVHA-aN771S and PlVHA-aN846S point mutations in resistant mutants were identified by gene sequencing analyses. These two point mutations were validated as contributors to resistance in P. litchii through genetic transformation and molecular docking.

荔枝是一种在世界范围内备受追捧的水果,由于荔枝霉病(主要由荔枝疫霉引起),荔枝面临着重大的产量挑战。氟吡唑内酯对荔枝霜枯病具有显著的抑制作用,可用于荔枝霜枯病的防治。虽然了解荔枝对氟哌啶酸的抗性至关重要,但对其风险和机制的研究仍然有限。本研究测定了125株荔枝假单胞菌对氟哌唑内酯的敏感性,平均EC50值为0.131±0.037 μg/mL。通过对杀菌剂的适应,获得了4个抗性因子超过600的抗性突变体,表明这些菌株具有较高的抗性水平。复合适应度指数分析表明,抗性突变体的生存适应度显著低于亲本菌株。交叉耐药试验显示,氟吡唑啉与其他不同作用方式的杀菌剂无交叉耐药。氟哌啶胺交叉耐药呈阳性。综合评价表明荔枝对氟哌啶酸产生耐药性的风险中等。通过基因测序分析,鉴定出耐药突变体PlVHA-aN771S和PlVHA-aN846S点突变。通过遗传转化和分子对接验证了这两个点突变对荔枝抗性的贡献。
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引用次数: 0
LWRR: Landscape of Wheat Rust Resistance towards practical breeding design. LWRR:面向实用育种设计的小麦抗锈景观。
Pub Date : 2025-04-14 DOI: 10.1007/s44154-025-00232-x
Jiwen Zhao, Haitao Dong, Jinyu Han, Jingrui Ou, Tiantian Chen, Yuze Wang, Shengjie Liu, Rui Yu, Weijun Zheng, Chunlian Li, Zhensheng Kang, Dejun Han, Qingdong Zeng, Xiaojie Wang, Shengwei Ma, Jianhui Wu
{"title":"LWRR: Landscape of Wheat Rust Resistance towards practical breeding design.","authors":"Jiwen Zhao, Haitao Dong, Jinyu Han, Jingrui Ou, Tiantian Chen, Yuze Wang, Shengjie Liu, Rui Yu, Weijun Zheng, Chunlian Li, Zhensheng Kang, Dejun Han, Qingdong Zeng, Xiaojie Wang, Shengwei Ma, Jianhui Wu","doi":"10.1007/s44154-025-00232-x","DOIUrl":"https://doi.org/10.1007/s44154-025-00232-x","url":null,"abstract":"","PeriodicalId":74874,"journal":{"name":"Stress biology","volume":"5 1","pages":"25"},"PeriodicalIF":0.0,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11996731/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144032001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Evolutionary characteristics, expression patterns of wheat receptor-like kinases and functional analysis of TaCrRLK1L16. 小麦受体样激酶TaCrRLK1L16的进化特征、表达模式及功能分析。
Pub Date : 2025-04-03 DOI: 10.1007/s44154-025-00215-y
Guosen Zhao, Shiao Qin, Zhimin Wei, Xingxuan Bai, Jia Guo, Zhensheng Kang, Jun Guo

Reverse genetics research in complex hexaploid wheat often encounters challenges in determining the priority of gene functional characterization. This study aims to systematically analyze the wheat (Triticum aestivum) receptor-like kinase (TaRLK) gene family and develop an effective strategy to identify key candidate genes for further investigation. We identified 3,424 TaRLKs using bioinformatics methods and analyzed the diverse and conserved evolutionary relationships of RLKs among Arabidopsis, rice and wheat. Based on publicly available and our laboratory's transcriptome data, we comprehensively analyzed the transcriptional expression patterns of TaRLKs in response to various stresses, particularly Puccinia striiformis f. sp. tritici (Pst). The TaCrRLK1L16, which is upregulated during Pst infection and triggered cell death in Nicotiana benthamiana, has been identified as a key candidate gene for further functional characterization. Furthermore, our results suggested that the transgenic wheat overexpressing TaCrRLK1L16 significantly enhanced resistance to Pst. This study will provide valuable insights into understanding the evolutionary characteristics and expression patterns of TaRLKs while offering a novel strategy for determining the priority of key candidate TaRLKs.

复杂六倍体小麦的反向遗传学研究在确定基因功能表征的优先级方面经常遇到挑战。本研究旨在对小麦(Triticum aestivum)受体样激酶(receptor-like kinase, TaRLK)基因家族进行系统分析,并建立有效的策略来鉴定关键候选基因,为进一步的研究奠定基础。利用生物信息学方法鉴定了3424个TaRLKs,并分析了拟南芥、水稻和小麦中RLKs的多样性和保守性进化关系。基于公开的转录组数据和我们实验室的转录组数据,我们全面分析了TaRLKs在各种胁迫下的转录表达模式,特别是小麦锈菌(Pst)。TaCrRLK1L16在Pst感染过程中上调,并引发本烟的细胞死亡,已被确定为进一步功能表征的关键候选基因。此外,我们的研究结果表明,过表达TaCrRLK1L16的转基因小麦显著增强了对Pst的抗性。该研究将为理解TaRLKs的进化特征和表达模式提供有价值的见解,同时为确定关键候选TaRLKs的优先级提供新的策略。
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引用次数: 0
A fungal effector hijacks a plastid protein to dampen plant immunity; PR1 is here for rescue. 真菌效应物劫持质体蛋白抑制植物免疫力;PR1是来救援的。
Pub Date : 2025-04-02 DOI: 10.1007/s44154-025-00230-z
Muhammad Saad Shoaib Khan, Faisal Islam, Huan Chen, Jian Chen

Plants are engaged in a constant battle for survival against pathogens, which triggers a multifaceted immune response characterized by pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) to prevent infection. These two immune responses operate synergistically to enhance plant immunity. PTI is considered the first line of defense involving the recognition of pathogen-associated molecular patterns (PAMPs) by specific receptors in host cells known as pattern recognition receptors (PRRs), which initiate defense signaling. However, many pathogens often overcome the first line of defense (PTI) and successfully deploy effector proteins to promote virulence and subvert plant immunity, leading to host susceptibility. In the counter-defense, the ETI defense mechanism is activated by triggering resistance (R) genes in plants that usually encode nucleotide-binding-leucine-rich-containing (NLR) proteins. During plant-pathogen interactions, transcriptional reprogramming of defense-related genes such as pathogenesis-related proteins and generation of reactive oxygen species (ROS) are essential for facilitating programmed cell death at the infected location to inhibit pathogen proliferation. While ROS and PR protein are critical in plant-pathogen interaction, they are not universally required or effective against all pathogens. Hence, plants' multilayer immune layer is encrypted with the compensatory activation of ETI defense response towards the failure of one component of the defense system to maintain robust immunity.

植物与病原体进行持续的生存斗争,从而引发以模式触发免疫(PTI)和效应触发免疫(ETI)为特征的多方面免疫反应来预防感染。这两种免疫反应协同作用,增强植物免疫力。PTI被认为是宿主细胞中被称为模式识别受体(PRRs)的特定受体识别病原体相关分子模式(PAMPs)的第一道防线,它启动防御信号。然而,许多病原体经常克服第一道防线(PTI)并成功地部署效应蛋白来增强毒力并破坏植物免疫,导致宿主易感性。在反防御中,ETI防御机制通过触发植物中通常编码核苷酸结合-富含亮氨酸(NLR)蛋白的抗性(R)基因而被激活。在植物与病原体相互作用过程中,防御相关基因(如致病相关蛋白)的转录重编程和活性氧(ROS)的产生对于促进感染部位的程序性细胞死亡以抑制病原体增殖至关重要。虽然ROS和PR蛋白在植物与病原体的相互作用中起着至关重要的作用,但它们并不是普遍需要的,也不是对所有病原体都有效。因此,植物的多层免疫层被ETI防御反应的补偿性激活加密,以应对防御系统中一个组成部分的失败,以维持强大的免疫力。
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引用次数: 0
One stone for multiple birds: PigmR integrates multiple defense pathways for high and broad-spectrum blast resistance in rice. 一石多鸟:PigmR整合了多种防御途径,用于水稻的高谱和广谱抗稻瘟病。
Pub Date : 2025-03-31 DOI: 10.1007/s44154-025-00228-7
Zhuoer Xie, Leiyun Yang, Zhengguang Zhang
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引用次数: 0
Genetic characterization of the AHAS mutant line K4 with resistance to AHAS-inhibitor herbicides in rapeseed (Brassica napus L.). 甘蓝型油菜抗AHAS除草剂突变系K4的遗传特性
Pub Date : 2025-02-25 DOI: 10.1007/s44154-024-00184-8
Yani Zhang, Qianxin Huang, Shengnan Wang, Lianliang Gao, Gaoping Qu, Yuan Guo, Zhaoxin Hu, Shengwu Hu

It remains a great challenge to control weeds in rapeseed fields in China. Breeding herbicide-resistant rapeseed varieties and using corresponding herbicide formulations has become the most economical and effective way to control weeds in rapeseed field. Characterization of more herbicide-resistant genetic resources will provide opportunities for breeders to develop rapeseed herbicide-resistant varieties with good agronomic performance. Previously, we obtained the tribenuron methyl (TBM)-resistant mutant K4 from ZS9 (Brassica napus L.) through ethyl methyl sulfonate mutagenesis and TBM foliar-spray screening. In this study, the inheritance and molecular characterization of the mutant K4 are carried out. Genetic investigation indicated that the herbicide-resistance of the K4 was controlled by one dominant allele at a single nuclear gene locus. Molecular characterization showed that a single point substitution at position 535 from C to T in BnAHAS3 (BnAHAS3535T), which resulted in a mutation at point 179 in BnAHAS3. The K4 showed a certain degree of resistance to TBM, bensulfuron methyl, and monosulfon sodium, which were 50, 30, and 5 times that of ZS9, respectively. AHAS enzyme assay, structural analysis of AHAS proteins, affinity detection between TBM and BnAHAS3 by surface plasmon resonance analysis, and the transgenic experiment in Arabidopsis using BnAHAS3535T confirmed that BnAHAS3535T endow the K4 with herbicides resistance. In addition, an allele-specific marker was developed to quickly distinguish the heterozygous and homozygous mutated alleles BnAHAS3535T. In conclusion, our research identified and characterized one novel mutative AHAS allele in B. napus and enriched genetic resource for developing herbicide-resistant rapeseed cultivars.

中国油菜地杂草的防治仍然是一个巨大的挑战。培育抗除草剂油菜籽品种并使用相应的除草剂配方已成为防治油菜田杂草最经济有效的途径。更多抗除草剂遗传资源的鉴定将为育种者培育农艺性能良好的油菜抗除草剂品种提供机会。此前,我们通过甲基磺酸乙酯诱变和TBM叶面喷雾筛选,从甘蓝型油菜ZS9 (Brassica napus L.)中获得了抗甲基三苯脲(tribenuron methyl, TBM)突变体K4。本研究对突变体K4的遗传和分子特性进行了研究。遗传分析表明,K4的抗除草剂性由单个核基因位点上的一个显性等位基因控制。分子鉴定表明,BnAHAS3基因在C到T的535位点发生单点取代(BnAHAS3535T),导致BnAHAS3基因在179位点发生突变。K4对TBM、甲苯磺隆和单磺隆钠均有一定的抗性,分别是ZS9的50倍、30倍和5倍。AHAS酶分析、AHAS蛋白结构分析、表面等离子体共振分析检测TBM与BnAHAS3的亲和性,以及BnAHAS3535T转基因拟南芥实验证实,BnAHAS3535T赋予K4抗除草剂能力。此外,开发了等位基因特异性标记,快速区分杂合和纯合突变等位基因BnAHAS3535T。总之,本研究鉴定并鉴定了甘蓝型油菜中一个新的AHAS突变等位基因,为开发抗除草剂油菜品种提供了丰富的遗传资源。
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引用次数: 0
Candidate genes associated with low temperature tolerance in cucumber adult plants identified by combining GWAS & QTL mapping. 利用GWAS和QTL定位相结合的方法鉴定黄瓜成株耐低温相关候选基因。
Pub Date : 2024-12-11 DOI: 10.1007/s44154-024-00191-9
Caixia Li, Shaoyun Dong, Diane M Beckles, Xiaoping Liu, Jiantao Guan, Zaizhan Wang, Xingfang Gu, Han Miao, Shengping Zhang

Fruit quality and yield are reduced when cucumber (Cucumis sativus L.) plants are exposed to low temperature (LT) stress, yet, the inheritance and genes linked to cold tolerance in adult plants have not been reported yet. Here, the LT-tolerance of 120 cucumber accessions representing four ecotypes were evaluated by GWAS, and also, in 140 recombinant inbred lines (RILs) derived from a biparental cross. Plants were exposed to naturally occurring LT environments in a plastic greenhouse, in winter 2022, and 2023, and a low temperature injury index (LTII) was employed to evaluate plant performance. Genetic analysis revealed that the LT-tolerance evaluated in the adult cucumber plants was a multigenic quantitative trait, and that 18 of the 120 accessions were highly LT tolerant by our LTII assessment. Two loci (gLTT1.1 and gLTT3.1) exhibited strong signals that were consistent and stable in two environments. In addition, two QTLs-qLTT1.2 on chromosome (Chr.) 1, and qLTT3.1 on Chr. 3, were discovered in all tests using RIL population derived from a cross between LT-sensitive 'CsIVF0106', and LT-tolerant 'CsIVF0168'. qLTT1.2 was delimited to a 1.24-Mb region and qLTT3.1 was narrowed to a 1.43-Mb region. Interestingly, a peak single nucleotide polymorphism (SNP) at gLTT1.1 and gLTT3.1 was also found in qLTT1.2 and qLTT3.1, respectively. These loci were thus renamed as gLTT1.1 and gLTT3.1. In these regions, 25 genes were associated with the LT response. By identifying differences in haplotypes and transcript profiles among these genes, we identified four candidates: CsaV3_1G012520 (an ethylene-responsive transcription factor) and CsaV3_1G013060 (a RING/U-box superfamily protein) in gLTT1.1, and two RING-type E3 ubiquitin transferases at CsaV3_3G018440 and CsaV3_3G017700 in gLTT3.1 that may regulate LT-tolerance in adult cucumber. Interestingly, the accessions in which the LT-tolerant haplotypes for two loci were pyramided, displayed maximally high tolerance for LT. These findings therefore provide a solid foundation for the identification of LT-tolerant genes and the molecular breeding of cucumber with LT-tolerance.

黄瓜(Cucumis sativus L.)在低温胁迫下果实品质和产量下降,但成株耐冷性的遗传和相关基因尚未见报道。本研究利用GWAS技术对来自4个生态型的120个黄瓜材料和140个双亲本杂交的重组自交系(RILs)的耐低温性进行了评价。将植物分别于2022年和2023年冬季暴露于塑料大棚的低温环境中,采用低温伤害指数(LTII)评价植物的低温性能。遗传分析表明,成体黄瓜耐盐性是一种多基因数量性状,经LTII鉴定,120份材料中有18份具有高耐盐性。两个基因座(gLTT1.1和gLTT3.1)在两种环境中表现出一致和稳定的强信号。此外,在使用由lt敏感的‘CsIVF0106’和lt耐受性的‘CsIVF0168’杂交而来的RIL群体进行的所有测试中,都发现了两个qtl - qltt1.2位于染色体(Chr.) 1, qLTT3.1位于Chr. 3。qLTT1.2被划分为1.24 mb的区域,qLTT3.1被缩小到1.43 mb的区域。有趣的是,gLTT1.1和gLTT3.1的单核苷酸多态性(SNP)峰值也分别在qLTT1.2和qLTT3.1中被发现。这些基因座因此被重命名为gLTT1.1和gLTT3.1。在这些区域中,有25个基因与LT反应相关。通过鉴定这些基因之间的单倍型和转录谱的差异,我们确定了四个候选基因:gLTT1.1中的CsaV3_1G012520(乙烯应答转录因子)和CsaV3_1G013060 (RING/U-box超家族蛋白),以及gLTT3.1中的CsaV3_3G018440和CsaV3_3G017700两个RING型E3泛素转移酶,它们可能调节成年黄瓜的耐盐性。有趣的是,具有两个位点的耐lt单倍型的材料对lt的耐受性最高。这些发现为耐lt基因的鉴定和耐lt黄瓜的分子育种提供了坚实的基础。
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