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A transcription factor, PbWRKY24, contributes to russet skin formation in pear fruits by modulating lignin accumulation 转录因子 PbWRKY24 通过调节木质素积累促进梨果赤褐色果皮的形成
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-18 DOI: 10.1093/hr/uhae300
Jialong Wang, Dong Wang, Mingrui Zhao, Mengyuan Yu, Xiaodong Zheng, Yike Tian, Zhijuan Sun, Xiaoli Liu, Caihong Wang, Changqing Ma
Skin color is one of the major traits of fruit appearance quality in pear (Pyrus), which affects the fruit commodity value. Russet skin protects pear fruits from environmental stresses and its formation process is closely linked to lignin accumulation. However, the molecular regulatory networks underlying russet skin formation in pear fruits involve complex secondary metabolic pathways and remain elusive. Here, we explored the regulatory mechanisms underlying lignin accumulation in pear skin based on transcriptome sequencing, co-expression network analysis, and gene expression profiling. We identified a WRKY transcription factor gene, PbWRKY24, that regulates russet skin formation in pear fruits. The relative expression of PbWRKY24 in russet pear skin was significantly correlated with lignin content. We then verified the function of PbWRKY24 in lignin accumulation via genetic transformation. DNA affinity purification sequencing revealed that PbWRKY24 directly binds to the promoter of a lignin biosynthesis gene, PbPRX4. This binding was confirmed by yeast one-hybrid, dual-luciferase, and electrophoretic mobility shift assays. Overexpression of PbPRX4 in pear skin stimulated lignin accumulation and consequently promoted russet skin formation. This study provides a glimpse into the intricate lignin biosynthesis mechanisms during russet skin formation in pear fruits, which is of practical significance to pear breeding for fruit quality.
果皮颜色是梨果实外观品质的主要特征之一,影响果实的商品价值。赤褐色果皮保护梨果免受环境胁迫,其形成过程与木质素积累密切相关。然而,梨果实中赤褐色果皮形成的分子调控网络涉及复杂的次级代谢途径,仍然难以捉摸。在此,我们基于转录组测序、共表达网络分析和基因表达谱分析,探索了梨皮中木质素积累的调控机制。我们发现了一个 WRKY 转录因子基因 PbWRKY24,它调控梨果实赤褐色果皮的形成。PbWRKY24 在赤皮梨果皮中的相对表达量与木质素含量显著相关。我们随后通过基因转化验证了 PbWRKY24 在木质素积累中的功能。DNA 亲和纯化测序显示,PbWRKY24 直接与木质素生物合成基因 PbPRX4 的启动子结合。酵母单杂交、双荧光素酶和电泳迁移试验证实了这种结合。在梨皮中过表达 PbPRX4 会刺激木质素的积累,从而促进赤褐色果皮的形成。这项研究揭示了梨果实赤褐色果皮形成过程中复杂的木质素生物合成机制,对梨果实品质育种具有重要的现实意义。
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引用次数: 0
Fertility restorer gene CaRf and PepperSNP50K provide a promising breeding system for hybrid pepper 生育力恢复基因 CaRf 和 PepperSNP50K 为杂交辣椒提供了一个前景广阔的育种系统
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-16 DOI: 10.1093/hr/uhae223
Bingqian Tang, Huiping Yang, Qinbiao Yin, Wu Miao, Yuting Lei, Qingzhi Cui, Jiawen Cheng, Xinhao Zhang, Ying Chen, Juan Du, Lingling Xie, Shunxue Tang, Meiqi Wang, Jiayue Li, Mingyue Cao, Li Chen, Fangling Xie, Xiumin Li, Fan Zhu, Zhongyi Wang, Cheng Xiong, Xiongze Dai, Xuexiao Zou, Feng Liu
Cytoplasmic male sterility (CMS) is pivotal in plant breeding and widely employed in various crop hybrids, including pepper. However, the functional validation of the restorer of fertility (Rf) gene in pepper has been lacking until now. This study identifies and characterizes CaRf, a single dominant locus crucial for restoring CMS in the pepper strong recovery inbred line Zhangshugang. The CaRf gene encodes a mitochondria-targeted pentatricopeptide repeat protein, validated through the induction of male sterility upon its silencing in hybrid F1 plants. To enhance pepper breeding efficiency, 176 important pepper breeding parent materials were resequenced, and a PepperSNP50K liquid-phase breeding chip was developed, comprising 51 172 markers. Integration of CaRf functional characterization and PepperSNP50K facilitated the development of a high-quality red pepper hybrid. These findings provide significant insights and practical strategies for advancing molecular-designed breeding in peppers.
细胞质雄性不育(CMS)在植物育种中至关重要,被广泛应用于包括辣椒在内的各种作物杂交种中。然而,迄今为止还缺乏对辣椒育性恢复基因(Rf)的功能验证。本研究鉴定并描述了辣椒强恢复性近交系樟树岗(Zhangshugang)中对恢复CMS至关重要的单显性基因座CaRf。CaRf 基因编码一种线粒体靶向五肽重复蛋白,通过在杂交 F1 植株中沉默 CaRf 基因诱导雄性不育得到了验证。为提高辣椒育种效率,对 176 个重要的辣椒育种亲本进行了重新测序,并开发了由 51 172 个标记组成的 PepperSNP50K 液相育种芯片。CaRf功能表征与PepperSNP50K的整合促进了优质红辣椒杂交种的培育。这些发现为推进辣椒分子设计育种提供了重要见解和实用策略。
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引用次数: 0
Diversity and interactions of rhizobacteria determine multinutrient traits in tomato host plants under nitrogen and water disturbances 根瘤菌的多样性和相互作用决定了氮和水干扰下番茄寄主植物的多营养性状
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-15 DOI: 10.1093/hr/uhae290
Wenxuan Shi, Junjie Guo, Xinxuan Yu, Zhixing Li, Boyang Weng, Danxia Wang, Shihao Su, Yufei Sun, Jinfang Tan, Ruohan Xie
Coevolution within the plant holobiont extends the capacity of host plants for nutrient acquisition and stress resistance. However, the role of the rhizospheric microbiota in maintaining multinutrient utilization (i.e., multinutrient traits) in the host remains to be elucidated. Multinutrient cycling index (MNC), analogous to the widely used multifunctionality index, provides a straightforward and interpretable measure of the multinutrient traits in host plants. Using tomato as a model plant, we characterized MNC (based on multiple aboveground nutrient contents) in host plants under different nitrogen and water supply regimes and explored the associations between rhizospheric bacterial community assemblages and host-plant multinutrient profiles. Rhizosphere bacterial community diversity, quantitative abundance, predicted function, and key topological features of the co-occurrence network were more sensitive to water supply than to nitrogen supply. A core bacteriome comprising 61 genera, such as Candidatus Koribacter and Streptomyces, persisted across different habitats and served as a key predictor of host-plant nutrient uptake. The MNC index increased with greater diversity and higher core taxon abundance in the rhizobacterial community, while decreasing with higher average degree and graph density of rhizobacterial co-occurrence network. Multinutrient absorption by host plants was primarily regulated by community diversity and rhizobacterial network complexity under the interaction of nitrogen and water. The high biodiversity and complex species interactions of the rhizospheric bacteriome play crucial roles in host-plant performance. This study supports the development of rhizosphere microbiome engineering, facilitating effective manipulation of the microbiome for enhanced plant benefits, which supports sustainable agricultural practices and plant health.
植物全生物体内的共同进化提高了宿主植物获取养分和抗逆的能力。然而,根瘤微生物群在维持宿主多养分利用(即多养分性状)方面的作用仍有待阐明。多养分循环指数(MNC)类似于广泛使用的多功能指数,它提供了一种直接且可解释的宿主植物多养分性状测量方法。我们以番茄为模式植物,描述了不同氮和水供应制度下寄主植物的多营养元素循环指数(基于多种地上营养元素含量),并探讨了根瘤菌群落组合与寄主植物多营养元素特征之间的关系。根圈细菌群落的多样性、定量丰度、预测功能以及共生网络的关键拓扑特征对供水的敏感性高于对供氮量的敏感性。由 61 个菌属组成的核心细菌群(如 Candidatus Koribacter 和 Streptomyces)在不同生境中持续存在,是预测寄主植物养分吸收的关键因素。MNC指数随着根瘤菌群落多样性和核心类群丰度的提高而增加,同时随着根瘤菌共生网络平均程度和图密度的提高而降低。在氮和水的相互作用下,寄主植物对多种营养元素的吸收主要受群落多样性和根瘤菌网络复杂性的调控。根瘤菌群的高度生物多样性和复杂的物种相互作用对寄主植物的表现起着至关重要的作用。这项研究支持了根圈微生物组工程学的发展,有助于有效操纵微生物组以提高植物效益,从而支持可持续农业实践和植物健康。
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引用次数: 0
Genome Editing for Grass Improvement and Future Agriculture 基因组编辑促进草地改良和未来农业
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-15 DOI: 10.1093/hr/uhae293
Bilal Muhammad, Jie Geng, Lin Chen, Pedro García-Caparros, Tao Hu
Grasses, including turf and forage, cover most of the earth’s surface; predominantly important for land, water, livestock feed, soil and water conservation, as well as carbon sequestration. Improved production and quality of grasses by modern molecular breeding is gaining more research attention. Recent advances in genome-editing technologies are helping to revolutionize plant breeding and also offering smart and efficient acceleration on grass improvement. Here, we reviewed all recent researches using (CRISPR)/CRISPR-associated protein (Cas)-mediated genome editing tools to enhance the growth and quality of forage and turf grasses. Furthermore, we highlighted emerging approaches aimed at advancing grass breeding program. We assessed the CRISPR-Cas effectiveness, discussed the challenges associated with its application, and explored future perspectives primarily focusing on turf and forage grasses. Despite the promising potential of genome editing in grasses, its current efficiency remains limited due to several bottlenecks, such as the absence of comprehensive reference genomes, the lack of efficient gene delivery tools, unavailability of suitable vector and delivery for grass species, high polyploidization, multiple homoeoalleles, etc. Despite these challenges, the CRISPR-Cas system holds great potential to fully harness its benefits in grass breeding and genetics, aiming to improve and sustain the quantity and quality of turf and forage grasses.
禾本科植物,包括草皮和牧草,覆盖了地球表面的大部分区域;对土地、水、牲畜饲料、水土保持以及碳固存具有重要意义。通过现代分子育种提高草的产量和质量正受到越来越多的研究关注。基因组编辑技术的最新进展有助于彻底改变植物育种,同时也为禾本科植物的改良提供了智能、高效的加速手段。在此,我们回顾了近期利用(CRISPR)/CRISPR 相关蛋白(Cas)介导的基因组编辑工具来提高牧草和草坪草生长和质量的所有研究。此外,我们还重点介绍了旨在推进牧草育种计划的新兴方法。我们评估了CRISPR-Cas的有效性,讨论了与应用CRISPR-Cas相关的挑战,并主要针对草皮和牧草探索了未来的前景。尽管基因组编辑在禾本科植物中的应用前景广阔,但目前的效率仍受到一些瓶颈的限制,如缺乏全面的参考基因组、缺乏高效的基因传递工具、没有适合禾本科植物的载体和传递方式、多倍体化程度高、多同源等。尽管存在这些挑战,CRISPR-Cas 系统仍具有巨大潜力,可充分利用其在草育种和遗传学方面的优势,提高并保持草坪和牧草的数量和质量。
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引用次数: 0
CsNAC17 enhances resistance to Colletotrichum gloeosporioides by interacting with CsbHLH62 in Camellia sinensis CsNAC17 通过与山茶中的 CsbHLH62 相互作用,增强山茶对球孢子菌的抗性
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-14 DOI: 10.1093/hr/uhae295
Rui Han, Huiling Mei, Qiwei Huang, Cunqiang Ma, Yuxin Zhao, Anbuaj Jeyaraj, Jing Zhuang, Yuhua Wang, Xuan Chen, Shujing Liu, Xinghui Li
The pathogen Colletotrichum gloeosporioides causes anthracnose, a serious threat to tea trees around the world, particularly in warm and humid regions. RNA-Seq data have previously indicated NAC transcription factors are involved in anthracnose resistance, but underlying mechanisms remain unclear. The BiFC, Split-LUC, and Co-IP assays validated the interaction between CsbHLH62 and CsNAC17 identified through yeast two-hybrid (Y2H) screening. CsNAC17 or CsbHLH62 overexpression enhanced anthracnose resistance, as well as enhanced levels of H2O2, hypersensitivity, and cell death in Nicotiana benthamiana. The NBS-LRR gene CsRPM1 is regulated by CsNAC17 by binding directly to its promoter (i.e., CACG, CATGTG), while CsbHLH62 facilitates CsNAC17’s binding and increases trascriptional activity of CsRPM1. Additionally, transient silencing of CsNAC17 and CsbHLH62 in tea plant leaves using the virus-induced gene silencing (VIGS) system resulted in decreased resistance to anthracnose. Conversely, transient overexpression of CsNAC17 and CsbHLH62 in tea leaves significantly enhanced the resistance against anthracnose. Based on these results, it appears that CsbHLH62 facilitates the activity of CsNAC17 on CsRPM1, contributing to increased anthracnose resistance.
病原体球孢子菌(Colletotrichum gloeosporioides)会引起炭疽病,严重威胁世界各地的茶树,尤其是温暖潮湿地区的茶树。之前的 RNA-Seq 数据表明,NAC 转录因子参与了炭疽病的抗性,但其潜在机制仍不清楚。BiFC、Split-LUC 和 Co-IP 试验验证了通过酵母双杂交(Y2H)筛选确定的 CsbHLH62 和 CsNAC17 之间的相互作用。CsNAC17 或 CsbHLH62 的过表达增强了烟草菌的抗炭疽病能力,同时也增强了烟草菌的 H2O2 水平、超敏性和细胞死亡。NBS-LRR 基因 CsRPM1 受 CsNAC17 的调控,直接与其启动子结合(即 CACG、CATGTG),而 CsbHLH62 可促进 CsNAC17 的结合并提高 CsRPM1 的转录活性。此外,利用病毒诱导基因沉默(VIGS)系统在茶树叶片中瞬时沉默 CsNAC17 和 CsbHLH62 会降低茶树对炭疽病的抗性。相反,CsNAC17 和 CsbHLH62 在茶叶中的瞬时过表达则显著增强了茶叶对炭疽病的抗性。基于这些结果,CsbHLH62 似乎促进了 CsNAC17 对 CsRPM1 的活性,从而提高了炭疽病的抗性。
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引用次数: 0
Tissue Culture-Independent Approaches to Revolutionizing Plant Transformation and Gene Editing 彻底改变植物转化和基因编辑的组织培养独立方法
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-14 DOI: 10.1093/hr/uhae292
Luis Felipe Quiroz, Moman Khan, Nikita Gondalia, Linyi Lai, Peter C McKeown, Galina Brychkova, Charles Spillane
Despite the transformative power of gene editing for crop improvement, its widespread application across species and varieties is limited by the transformation bottleneck that exists for many crops. The genetic transformation of plants is hindered by a general reliance on in vitro regeneration through plant tissue culture. Tissue culture requires empirically-determined conditions and aseptic techniques, and cannot easily be translated to recalcitrant species and genotypes. Both Agrobacterium-mediated and alternative transformation protocols are limited by a dependency on in vitro regeneration, which also limits their use by non-experts and hinders research into non-model species such as those of possible novel biopharmaceutical or nutraceutical use, as well as novel ornamental varieties. Hence, there is significant interest in developing tissue culture-independent plant transformation and gene editing approaches which can circumvent the bottlenecks associated with in vitro plant regeneration recalcitrance. Compared to tissue culture-based transformations, tissue culture-independent approaches offer advantages such as avoidance of somaclonal variation effects, with more streamlined and expeditious methodological processes. The ease of use, dependability, and accessibility of tissue culture-independent procedures can make them attractive to non-experts, outperforming classic tissue culture-dependent systems. This review explores the diversity of tissue culture-independent transformation approaches and compares them to traditional tissue culture-dependent transformation strategies. We highlight their simplicity and provide examples of recent successful transformations accomplished using these systems. Our review also addresses current limitations and explores future perspectives, highlighting the significance of these techniques for advancing plant research and crop improvement.
尽管基因编辑在作物改良方面具有强大的转化能力,但由于许多作物存在转化瓶颈,基因编辑在不同物种和品种间的广泛应用受到了限制。植物的基因转化普遍依赖于通过植物组织培养进行体外再生。组织培养需要经验确定的条件和无菌技术,不容易转化为难转化的物种和基因型。农杆菌介导的转化协议和替代转化协议都因依赖体外再生而受到限制,这也限制了非专业人员对它们的使用,并阻碍了对非模式物种的研究,例如那些可能具有新型生物制药或营养保健用途的物种,以及新型观赏品种。因此,人们对开发不依赖组织培养的植物转化和基因编辑方法产生了浓厚的兴趣,这种方法可以规避与体外植物再生顽固性相关的瓶颈。与基于组织培养的转化相比,独立于组织培养的方法具有避免体细胞变异效应等优势,而且方法流程更加简化和快捷。独立于组织培养的程序使用方便、可靠、易于获得,因此对非专业人员很有吸引力,优于传统的依赖组织培养的系统。本综述探讨了独立于组织培养的转化方法的多样性,并将其与传统的依赖组织培养的转化策略进行了比较。我们强调了它们的简便性,并提供了最近使用这些系统成功完成转化的实例。我们的综述还讨论了目前的局限性,并探讨了未来的前景,强调了这些技术对推动植物研究和作物改良的重要意义。
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引用次数: 0
CHH hypermethylation contributes to the early ripening of grapes revealed by DNA methylome landscape of “Kyoho” and its bud mutant 通过 "Kyoho "及其花蕾突变体的 DNA 甲基组图谱揭示 CHH 超甲基化对葡萄早熟的影响
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-14 DOI: 10.1093/hr/uhae285
Tong-Lu Wei, Yu-Tong Wan, Hai-Nan Liu, Mao-Song Pei, Guang-Qi He, Da-Long Guo
DNA methylation is a stable epigenetic mark that plays a crucial role in plant life processes. However, the specific functions of DNA methylation in grape berry development remain largely unknown. In this study, we performed whole-genome bisulfite sequencing on “Kyoho” grape and its early-ripening bud mutant “Fengzao” at different developmental stages. Our results revealed that transposons (TEs) and gene flanking regions exhibited high levels of methylation, particularly in “Fengzao”, attributed to CHH site methylation. Interestingly, the methylation patterns in these two cultivars showed distinct dynamics during berry development. While methylation levels of genes and TEs increased gradually in “Kyoho” throughout berry development, “Fengzao” did not display consistent changes. Notably, “Fengzao” exhibited higher methylation levels in promoters compared to “Kyoho”, suggesting that hypermethylation of promoters may contribute to its early ripening phenotype. Integration of methylome and transcriptome data highlighted differentially methylated genes (DMGs) and expressed genes (DEGs) associated with secondary metabolite biosynthesis, with 38 genes identified as potential candidates involved in grape berry development. Furthermore, the study identified a jasmonate-induced oxygenase gene (JOX1) as a negative regulator of ripening in Arabidopsis and grapes, indicating that hypermethylation of JOX1 may play a role in the early ripening of “Fengzao”. Overall, our findings provide insights into the distinct DNA methylation patterns during grape berry development, shedding light on the epigenetic regulatory mechanisms underlying the early-ripening bud mutant.
DNA 甲基化是一种稳定的表观遗传标记,在植物生命过程中起着至关重要的作用。然而,DNA甲基化在葡萄浆果发育过程中的具体功能仍不为人知。在这项研究中,我们对 "京丰 "葡萄及其早熟芽突变体 "丰早 "在不同发育阶段进行了全基因组亚硫酸氢盐测序。结果表明,转座子(TE)和基因侧翼区域表现出较高的甲基化水平,尤其是在 "丰早 "中,这归因于 CHH 位点的甲基化。有趣的是,这两个品种的甲基化模式在浆果发育过程中表现出不同的动态变化。在整个浆果发育过程中,"Kyoho "的基因和TEs甲基化水平逐渐增加,而 "Fengzao "则没有表现出一致的变化。值得注意的是,与 "Kyoho "相比,"Fengzao "的启动子甲基化水平更高,这表明启动子的超甲基化可能是其早熟表型的原因之一。甲基组和转录组数据的整合突显了与次生代谢物生物合成相关的差异甲基化基因(DMGs)和表达基因(DEGs),其中有 38 个基因被确定为参与葡萄浆果发育的潜在候选基因。此外,研究还发现茉莉酸诱导的加氧酶基因(JOX1)是拟南芥和葡萄成熟的负调控因子,这表明 JOX1 的高甲基化可能在 "凤枣 "的早熟过程中发挥作用。总之,我们的研究结果深入揭示了葡萄浆果发育过程中不同的DNA甲基化模式,揭示了早熟芽突变体的表观遗传调控机制。
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引用次数: 0
The amino acid permease SlAAP6 contributes to tomato growth and salt tolerance by mediating branched-chain amino acid transport 氨基酸渗透酶 SlAAP6 通过介导支链氨基酸转运促进番茄的生长和耐盐性
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-11 DOI: 10.1093/hr/uhae286
Qi Qiang, Zhonghui Zhang, Xianggui Li, Chun Li, Mengdi Mao, Xiangyu Ding, Jianing Zhang, Shixuan Li, Zesen Lai, Jie Yang, Peng Cao, Weizhen Ye, Shouchuang Wang, Jun Yang
Branched-chain amino acids (BCAAs) are essential amino acids in tomato (Solanum lycopersicum) required for protein synthesis, which also modulate growth and abiotic stress responses. To date, little is known about their uptake and transport in tomato especially under abiotic stress. Here, the tomato amino acid permease 6 (SlAAP6) gene was identified as an amino acid transporter that restored mutant yeast cell growth on media with a variety of amino acids, including BCAAs. Overexpression of SlAAP6 (SlAAP6-OE) in tomato raised the BCAA content and elevated the fresh weight, while SlAAP6 knockouts (slaap6) showed reduced levels of neutral and basic amino acids in seedling tissues and lower total free amino acid distribution to shoots. In comparison to wild type (WT) and slaap6 mutants, SlAAP6-OE alleviated root limited growth by elevated BCAA transport and upregulated the expression of root-growth-related genes by increasing BCAAs in vivo. As SlAAP6 serves as a positive regulator for BCAA abundance, SlAAP6-OE lines showed greater salinity tolerance, while slaap6 mutants exhibited increased salt sensitivity. The salt tolerance of SlAAP6-OE plants was further enhanced by the application of exogenous BCAAs. In addition, BCAA supplementation reduced the accumulation of H2O2 in root under salt stress conditions. Based on these findings, SlAAP6-mediated uptake and transport of BCAAs facilitated growth and salt tolerance in tomato. By characterizing this key amino acid transporter, this study provides a novel approach to simultaneously enhance tomato nutritional quality, growth and development, and stress resistance through genetic improvement.
支链氨基酸(BCAAs)是番茄(Solanum lycopersicum)蛋白质合成所需的必需氨基酸,也能调节生长和非生物胁迫反应。迄今为止,人们对其在番茄中的吸收和运输,尤其是在非生物胁迫下的吸收和运输知之甚少。在此,研究人员发现番茄氨基酸渗透酶 6(SlAAP6)基因是一种氨基酸转运体,它能恢复突变酵母细胞在含有多种氨基酸(包括 BCAAs)的培养基上的生长。在番茄中过表达 SlAAP6(SlAAP6-OE)可提高 BCAA 含量并增加鲜重,而 SlAAP6 基因敲除者(slaap6)则显示幼苗组织中的中性和碱性氨基酸含量降低,游离氨基酸在芽中的总分布降低。与野生型(WT)和slaap6突变体相比,SLAAP6-OE通过提高BCAA的运输来缓解根的生长受限,并通过增加体内的BCAA来上调根生长相关基因的表达。由于 SlAAP6 是 BCAA 丰度的正向调节因子,因此 SlAAP6-OE 株系表现出更强的耐盐性,而 slaap6 突变体则表现出更强的盐敏感性。施用外源 BCAAs 进一步增强了 SlAAP6-OE 植物的耐盐性。此外,在盐胁迫条件下,补充 BCAA 可减少根中 H2O2 的积累。基于这些发现,SlAAP6 介导的 BCAAs 吸收和转运促进了番茄的生长和耐盐性。通过表征这一关键氨基酸转运体,本研究为通过遗传改良同时提高番茄的营养品质、生长发育和抗逆性提供了一种新方法。
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引用次数: 0
Glycerophosphodiester phosphodiesterase 1 mediates G3P accumulation for Eureka lemon resistance to citrus yellow vein clearing virus 甘油磷酸二酯磷酸二酯酶 1 介导 G3P 积累,促进尤里卡柠檬对柑橘黄筋清病毒的抗性
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-11 DOI: 10.1093/hr/uhae287
Ping Liao, Xue Dan, Wen Ge, Qi Zhang, Jinfa Zhao, Changyong Zhou, Yan Zhou
Glycerophosphodiester phosphodiesterase 1 (GDPD1) plays an important function in the abiotic stress responses and participates in the accumulation of sn-glycerol-3-phosphate (G3P) in plants, which is key to plant systemic acquired resistance (SAR). However, the role of GDPD1 in plant responses to biotic stress remains poorly understood. This study characterized the antivirus function of the GDPD1 gene (designated as ClGDPD1) from Eureka lemon. ClGDPD1 is located in the membrane and endoplasmic reticulum, where it interacts with the citrus yellow vein clearing virus (CYVCV) coat protein (CP). Compared to individually express ClGDPD1 or co-expressed ClGDPD1 + CP140-326, transiently co-expressed ClGDPD1 + CP or ClGDPD1 + CP1-139 significantly up-regulated the key substance contents and genes expression involved in glycerophospholipid metabolism. Over-expression of ClGDPD1 significantly facilitated the accumulation of G3P, up-regulated the expression of SAR-related genes, and increased the resistance of transgenic Eureka lemon to CYVCV infection. Furthermore, exogenous glycerol treatment and over-expression of ClGPDH increased the G3P content and reduced CYVCV titers in plants or hairy roots. These results indicated that the enhanced resistance of ClGDPD1 transgenic Eureka lemon to CYVCV may be due to facilitating G3P accumulation through the interaction of ClGDPD1 with CP. Our findings provide novel insights into the role of ClGDPD1 as an important regulatory center in mediating the citrus defense response to viral infections.
甘油磷酸二酯磷酸二酯酶 1(GDPD1)在非生物胁迫响应中发挥着重要功能,它参与植物体内甘油-3-磷酸(G3P)的积累,是植物系统获得性抗性(SAR)的关键。然而,人们对 GDPD1 在植物生物胁迫响应中的作用仍然知之甚少。本研究鉴定了尤里卡柠檬中 GDPD1 基因(命名为 ClGDPD1)的抗病毒功能。ClGDPD1 位于膜和内质网中,与柑橘黄脉清病毒(CYVCV)的衣壳蛋白(CP)相互作用。与单独表达ClGDPD1或共表达ClGDPD1 + CP140-326相比,瞬时共表达ClGDPD1 + CP或ClGDPD1 + CP1-139能显著上调涉及甘油磷脂代谢的关键物质含量和基因表达。过度表达 ClGDPD1 能明显促进 G3P 的积累,上调 SAR 相关基因的表达,增强转基因尤里卡柠檬对 CYVCV 感染的抵抗力。此外,外源甘油处理和 ClGPDH 的过度表达增加了 G3P 含量,降低了植株或毛根中的 CYVCV 滴度。这些结果表明,ClGDPD1 转基因尤里卡柠檬对 CYVCV 的抗性增强可能是由于 ClGDPD1 与 CP 相互作用促进了 G3P 的积累。我们的研究结果为 ClGDPD1 作为一个重要的调控中心在介导柑橘对病毒感染的防御反应中的作用提供了新的见解。
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引用次数: 0
CRISPR/Cas9 mediated CHS2 mutation provides a new insight into resveratrol biosynthesis by causing a metabolic pathway shift from flavonoids to stilbenoids in Vitis davidii cells CRISPR/Cas9 介导的 CHS2 基因突变导致葡萄孢细胞中的代谢途径从黄酮类转变为二苯乙烯类,从而为白藜芦醇的生物合成提供了新的视角
IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences Pub Date : 2024-10-09 DOI: 10.1093/hr/uhae268
Gongti Lai, Peining Fu, Liyuan He, Jianmei Che, Qi Wang, Pufu Lai, Jiang Lu, Chengchun Lai
Resveratrol is an important phytoalexin that adapts to and responds to stressful conditions, plays various roles in health and medical therapies. However, it is only found in a limited number of plant species in low concentrations, which hinders its development and utilization. Chalcone synthase (CHS) and stilbene synthase (STS) catalyze the same substrates to produce flavonoids and resveratrol, respectively. However, it remains unclear how CHS and STS compete in metabolite synthesis. In this study, two CHS2 mutant cell lines (MT1 and MT2) were generated using CRISPR/Cas9 genome editing. These CHS2 mutant cell lines exhibited abundant mutations in CHS2, leading to the premature termination of protein translation and subsequent CHS2 knockout. Amplicon sequencing confirmed comprehensive CHS2 knockout in MT1, whereas the wild-type sequence remained predominant in the MT2 cell line. Transcriptome and RT-qPCR results showed a significant downregulation of genes involved in flavonoid biosynthesis, including CHS2, CHS3, F3H, F3’H, DFR, FLS, LDOX, among others, resulting in decreased flavonoid accumulation, such as anthocyanins, proanthocyanidins, quercetin, and kaempferol. Conversely, STS genes involved in stilbenoid biosynthesis were upregulated competing with the flavonoid pathway. Consequently, there was a marked increase in stilbenoids, including resveratrol, piceatannol, piceid and pterostilbene, with a 4.1-fold increase in resveratrol and a 5.3-fold increase in piceid (a derivative of resveratrol) observed in CHS2 mutant cell lines. This research demonstrates that CHS2 mutation induces a shift from flavonoid biosynthesis towards stilbenoid biosynthesis, offering new insights into metabolite biosynthesis and regulation, as well as an alternative solution for natural resveratrol production, and a novel breeding approach for eliminating non-target agronomic traits using CRISPR-Cas9.
白藜芦醇是一种重要的植物雌激素,能适应和应对压力条件,在健康和医学疗法中发挥着各种作用。然而,白藜芦醇只存在于数量有限、浓度较低的植物物种中,这阻碍了其开发和利用。查尔酮合成酶(CHS)和二苯乙烯合成酶(STS)催化相同的底物,分别产生类黄酮和白藜芦醇。然而,CHS 和 STS 如何在代谢物合成过程中竞争仍不清楚。本研究利用 CRISPR/Cas9 基因组编辑技术生成了两个 CHS2 突变细胞系(MT1 和 MT2)。这些CHS2突变细胞系中的CHS2发生了大量突变,导致蛋白质翻译过早终止,随后CHS2被敲除。扩增子测序证实了 MT1 中 CHS2 的全面基因敲除,而 MT2 细胞系中仍以野生型序列为主。转录组和 RT-qPCR 结果显示,参与黄酮类化合物生物合成的基因,包括 CHS2、CHS3、F3H、F3'H、DFR、FLS、LDOX 等,都出现了明显的下调,导致花青素、原花青素、槲皮素和山柰醇等黄酮类化合物的积累减少。相反,参与类芪生物合成的 STS 基因上调,与类黄酮途径竞争。因此,在 CHS2 突变体细胞系中,包括白藜芦醇、苦橙皮酚、苦橙皮苷和紫檀芪在内的类橙皮甙显著增加,其中白藜芦醇增加了 4.1 倍,苦橙皮苷(白藜芦醇的衍生物)增加了 5.3 倍。这项研究表明,CHS2 突变会诱导黄酮类生物合成向芪类生物合成转变,为代谢物的生物合成和调控提供了新的视角,也为天然白藜芦醇的生产提供了另一种解决方案,并为利用 CRISPR-Cas9 消除非目标农艺性状提供了一种新的育种方法。
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Horticulture Research
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