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Fermented polyherbal formulation ameliorates the severity of acute multiple - antibiotics - resistant Pseudomonas aeruginosa infected burn wound in rat burn model 复方发酵制剂可改善大鼠烧伤模型急性多重耐药铜绿假单胞菌感染创面的严重程度
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.13.566953
Subhanil Chakraborty, Subhajit Sen, Ranadhir Chakraborty
Pseudomonas aeruginosa, a Gram-negative opportunistic bacterium has emerged as a cause of life-threatening infections in topical burn wounds. Current therapeutic approaches through wound dressings and systemic medicines are far from satisfactory; multiple-antibiotic-resistance shown by pathogens contribute to failures of therapy causing mortality. This animal study was conducted to check efficacy of one Ayurveda based fermented polyherbal preparation (AP 01) against multiple antibiotic resistant Pseudomonas aeruginosa HW01 infected rat burn wounds. AP-01 was applied on artificially inflected burn wound on rat model infected with Multiple Antibiotics Resistant Pseudomonas aeruginosa to register the healing effects in terms of reduction in residual wound area percentage, presence of C-reactive protein in blood, presence of viable bacteria colony while keeping conventional antibiotics as positive control.The polyherbal preparation had reduced the infected residual burn wound area at 40.63% from the initial burn wound area within two weeks after a single intervention; whereas residual burn wound area remained much higher in case of animals left untreated and in case of the animals treated with control drug. Restoration to normalcy of serum C-reactive protein level were also achieved earlier in case of polyherbal AP-01 treated groups than other groups. Fermented formulation using components of AP-01 singly or in different combinations were never been tested earlier for topical application in infected burn-wound. The formulation AP-01 was found superior in terms of rate of healing and control of infection by multiple-antibiotic-resistant P. aeruginosa strains in burn wounds in rat model.
铜绿假单胞菌是一种革兰氏阴性机会性细菌,已成为局部烧伤创面危及生命的感染的原因。目前通过伤口敷料和全身药物的治疗方法远不能令人满意;病原体显示的多种抗生素耐药性导致治疗失败,导致死亡。本实验采用阿育吠陀发酵多药制剂(AP 01)对多重耐药铜绿假单胞菌(HW01)感染大鼠烧伤创面的疗效进行了实验研究。将AP-01应用于多种耐药铜绿假单胞菌感染大鼠模型人工感染创面,在常规抗生素为阳性对照的情况下,观察创面残留面积百分率、血c反应蛋白含量、活菌菌落存在等方面的愈合效果。在单次干预后两周内,多药制剂使感染的烧伤残余创面面积比初始创面面积减少40.63%;然而,在未治疗的动物和使用对照药物的动物的情况下,烧伤残余创面面积仍然要高得多。多药AP-01治疗组血清c反应蛋白水平恢复正常的时间也早于其他治疗组。使用AP-01成分单独或不同组合的发酵制剂在感染烧伤创面的局部应用方面从未进行过早期试验。结果表明,AP-01在大鼠模型烧伤创面的治愈率和多重耐药铜绿假单胞菌感染控制方面具有较好的效果。
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引用次数: 0
Emergence of the Novel Infectious bursal disease viruse Variant in Vaccinated Poultry Flocks in Egypt 在埃及接种疫苗的家禽中出现了新的传染性法氏囊病病毒变体
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.13.566865
Momtaz A Shahin, Hesham A Sultan, Ali Zanaty, Zienab Mosaad, Naglaa M Hagage, Amany Adel, Karim Selim, Dalia Saied, Abdel Hafez Samir, Ahmed Erfan, Abdelsatar Arafa, Abdullah Selim, Mohammed Samy, Mahmoud M Naguib, Heba Hassan, Osama El Shazly, Zeinab A Elbade, Mahmoud Kamel, Eman Farghaly, Samah Eid, Mohamed El Shahaby, Mohamed Hammoda, Osama Mehana, Mohamed Nabeh, Ahmed Samy
Infectious bursal disease viruses (IBDVs) have a profound impact on poultry production worldwide, directly causing mortality rates of up to 100%, and indirectly through their immunosuppressive effects. Since the emergence of the antigenically modified very virulent IBDV (vvIBDV) in Egypt in late 1999, the country has experienced recurrent outbreaks with high mortality rates and typical vvIBDV gross lesions. However, a notable shift occurred in 2023, characterized by a substantial increase in reported subclinical IBDV cases exhibiting atrophied bursa and associated immunosuppression. To assess the field situation, we examined samples from 21 farms in 2023 and 18 farms from 2021 and 2022, all of which experienced IBD outbreaks based on clinical diagnosis. These samples were submitted to our laboratory for confirmatory testing and subsequently subjected to VP2-HVR sequencing. Phylogenetic analysis revealed that all samples collected in 2021 and 2022 clustered with classical virulent strains and very virulent IBDV. In 2023, one sample clustered with the Egyptian vvIBDV, while one sample clustered with classic virulent IBDV, and the remaining 2023 samples clustered with the Chinese novel variant IBDV (nVarIBDV). The alignment of deduced amino acid sequences for VP2 revealed that all Egyptian classic virulent strains were similar to the Winterfield or Leukert strains. In contrast, vvIBDV strains exhibited two out of the three typical residues found in Egyptian antigenically atypical vvIBDV, namely Y220F and G254S, but not A321T, and one sample was identical to the European vvIBDV (emerged in 1989). Meanwhile, all variant strains recognized in the present study exhibited typical residues found in variant IBDV, in addition to the three conserved amino acid residues found only in Chinese variant IBDVs. However, all Egyptian variant strains showed a mutation at position 321 (321V), which represents the most exposed part of the capsid and is known to have a massive impact on IBDV antigenicity, with the exception of one sample that had 318G instead. This report highlights the emergence of a new variant IBDV clustered with the Chinese new variant in Egypt, causing bursa atrophy and spreading subclinically in broiler farms over a wide geographic distance, resulting in massive economic losses due to immunosuppression.
传染性法氏囊病病毒(IBDVs)对全世界的家禽生产有着深远的影响,直接导致高达100%的死亡率,并通过其免疫抑制作用间接造成死亡率。自从1999年底在埃及出现经过抗原性修饰的非常强毒的IBDV (vvIBDV)以来,该国经历了高死亡率和典型的vvIBDV大体病变的反复暴发。然而,2023年发生了显著的变化,其特征是报告的亚临床IBDV病例大幅增加,表现为囊萎缩和相关的免疫抑制。为了评估现场情况,我们检查了2023年21个农场和2021年和2022年18个农场的样本,根据临床诊断,这些农场都经历了IBD暴发。这些样本被提交到我们的实验室进行确认测试,随后进行VP2-HVR测序。系统发育分析显示,2021年和2022年收集的所有样本均聚集了经典强毒株和极强毒株。2023年,1份样本聚集为埃及型vvIBDV, 1份样本聚集为经典毒力型IBDV,其余2023份样本聚集为中国新型IBDV (nVarIBDV)。VP2的氨基酸序列比对表明,所有埃及经典毒力菌株与Winterfield或Leukert菌株相似。相比之下,vvIBDV株在埃及抗原性非典型vvIBDV中发现的三个典型残基中有两个,即Y220F和G254S,但没有A321T,并且一个样本与欧洲vvIBDV(1989年出现)相同。与此同时,本研究识别的所有变异菌株除了在中国变异IBDV中发现的3个保守氨基酸残基外,均具有变异IBDV的典型残基。然而,所有埃及变种菌株在321 (321V)位置显示突变,这是衣壳最暴露的部分,已知对IBDV抗原性有巨大影响,除了一个样本有318G。本报告强调了在埃及出现的与中国新变体聚集的新型IBDV,引起囊萎缩并在广泛的地理距离上亚临床地在肉鸡养殖场传播,由于免疫抑制导致巨大的经济损失。
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引用次数: 0
The heparin-binding domain of VEGF165 directly binds to integrin αvβ3 and plays a critical role in signaling. VEGF165的肝素结合域直接与整合素αvβ3结合,在信号传导中起关键作用。
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.14.567104
Yoko K Takada, Jessica Yu, Xiaojin Ye, Chun-Yi Wu, Brunie H Felding, Masaaki Fujita, yoshikazu takada
VEGF-A is a key cytokine in tumor angiogenesis and a major therapeutic target for cancer. VEGF165 is the predominant isoform and is the most potent angiogenesis stimulant. VEGFR2/KDR domains 2 and 3 (D2D3) bind to the N-terminal domain (NTD, residues 1-110) of VEGF165. Since removal of the heparin-binding domain (HBD, residues 111-165) markedly reduced the mitogenic activity of VEGF165, it has been proposed that the HBD plays a critical role in the mitogenicity of VEGF165. Integrin αvβ3 has been shown to bind to VEGF165, but the role of integrin αvβ3 in VEGF165 signaling are unclear. Here we describe that αvβ3 specifically bound to the isolated HBD, but not to the NTD. We identified several critical amino acid residues in HBD for integrin binding (Arg-123, Arg-124, Lys-125, Lys-140, Arg-145, and Arg-149) by docking simulation and mutagenesis, and generated full-length VEGF165 that is defective in integrin binding by including mutations in the HBD. The full-length VEGF165 mutant defective in integrin binding (R123A/R124A/K125A/K140A/R145A/R149A) was defective in ERK1/2 phosphorylation, integrin β3 phosphorylation, and KDR phosphorylation, although the mutation did not affect KDR binding to VEGF165. We propose a model in which VEGF165 induces KDR (through NTD)-VEGF165 (through HBD)-integrin αvβ3 ternary complex formation on the cell surface and this process is critically involved in potent mitogenicity of VEGF165.
VEGF-A是肿瘤血管生成的关键细胞因子,也是肿瘤的主要治疗靶点。VEGF165是主要的异构体,是最有效的血管生成兴奋剂。VEGFR2/KDR结构域2和3 (D2D3)与VEGF165的n端结构域(NTD,残基1-110)结合。由于去除肝素结合结构域(HBD,残基111-165)显著降低了VEGF165的有丝分裂活性,因此有人提出HBD在VEGF165的有丝分裂性中起关键作用。整合素αvβ3已被证实与VEGF165结合,但整合素αvβ3在VEGF165信号传导中的作用尚不清楚。在这里,我们描述了αvβ3特异性结合到分离的HBD,而不是NTD。通过对接模拟和诱变,我们确定了HBD中整合素结合的几个关键氨基酸残基(Arg-123、Arg-124、Lys-125、Lys-140、Arg-145和Arg-149),并通过在HBD中加入突变,生成了全长的整合素结合缺陷VEGF165。全长突变体VEGF165整合素结合缺陷(R123A/R124A/K125A/K140A/R145A/R149A)在ERK1/2磷酸化、整合素β3磷酸化和KDR磷酸化方面存在缺陷,尽管该突变不影响KDR与VEGF165的结合。我们提出了一个模型,其中VEGF165诱导KDR(通过NTD)-VEGF165(通过HBD)-整合素αvβ3三元复合物在细胞表面形成,这一过程对VEGF165的强有丝分裂性至关重要。
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引用次数: 0
Transcriptomic landscape ofseedstickinArabidopsis thalianafuniculus after fertilisation 拟南芥受精后种子粘附的转录组景观
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.13.566818
Maria João Ferreira, Jessy Silva, Hidenori Takeuchi, Takamasa Suzuki, Tetsuya Higashiyama, Sílvia Vieira de Almeida Coimbra
In Angiosperms, the continuation of plant species is intricately dependent on the funiculus multifaceted role in nutrient transport, mechanical support, and dehiscence of seeds. SEEDSTICK (STK) is a MADS-box transcription factor involved in seed size and dehiscence, and one of the few genes identified as affecting funiculus growth. Given the importance of the funiculus to a correct seed development, allied with previous phenotypic observations of stk mutants, we performed a transcriptomic analysis of stk funiculi, using RNA-sequencing, to infer on the deregulated networks of genes. The generated dataset of differentially expressed genes was enriched with cell wall biogenesis, cell cycle, sugar metabolism and transport terms, all in accordance with stk phenotype. We selected eight differentially expressed genes involved with abscission, seed development or novel functions in stk funiculus, such as hormones/secondary metabolites transport, for transcriptome validation using qPCR and/or promoter reporter lines. Overall, the analysis performed in this study allowed delving into the STK-network established in Arabidopsis funiculus, fulfilling a literature gap. Simultaneously, our findings reinforced the reliability of the transcriptome, and identified processes and new candidate genes that will enable a better understanding on the role of this sporophytic structure and how seed development may be affected by it.
在被子植物中,植物物种的延续复杂地依赖于球囊在营养转运、机械支持和种子开裂等方面的多重作用。SEEDSTICK (STK)是一种MADS-box转录因子,与种子大小和开裂有关,是为数不多的影响球茎生长的基因之一。考虑到索柄对正确的种子发育的重要性,结合之前对索柄突变体的表型观察,我们使用rna测序对索柄进行了转录组学分析,以推断基因的非调控网络。生成的差异表达基因数据集丰富了细胞壁生物发生、细胞周期、糖代谢和运输等术语,这些术语都与stk表型一致。我们选择了8个差异表达的基因,这些基因与茎的脱落、种子发育或新功能有关,如激素/次生代谢物运输,使用qPCR和/或启动子报告系进行转录组验证。总的来说,本研究的分析可以深入研究拟南芥中建立的stk -网络,填补了文献空白。同时,我们的发现加强了转录组的可靠性,并确定了过程和新的候选基因,这将使我们更好地了解这种孢子体结构的作用以及它如何影响种子发育。
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引用次数: 0
Comparing hormone dynamics in cereal crops via transient expression of hormone sensors 通过激素传感器的瞬时表达比较谷类作物的激素动态
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.14.567063
Thai Q Dao, Colleen Drapek, Alexander M Jones, Samuel Leiboff
Plant hormones are small molecules which elicit profound physiological responses. Although plant hormone biosynthesis and response genes have been critical for agricultural improvement, it has been difficult to experimentally compare hormone biology across species because of complex phenotypic outputs. We used transient expression of genetic hormone sensors and transcriptomics to quantify tissue-specific gibberellic acid (GA) and auxin responses across tissues and genotypes in cereal crops. We found that the FRET-based GPS2 biosensor detects exogenous GA treatments in maize, barley, sorghum, and wheat, in both vegetative and floral tissues. Measuring GPS2 output across GA dosages revealed tissue- and genotype-specific differences in GA sensor response. We observed marked differences in maize vs barley leaves and floral tissues and an unexpected drop in GPS2 output in the maize d1 GA biosynthesis mutant after GA treatment, likely reflecting differences in bioactive GA content, GA transport, and mechanisms of GA response. We then used RNAseq to measure transcriptional responses to GA treatment in leaves from maize wildtype, d1, and barley as well as floral tissues from maize and barley for a cross-tissue, cross-genotype, and cross-species GA-response comparison. After orthology prediction and analysis of within- and cross-species GO-term enrichment, we identified core sets of GA-responsive genes in each species as well as maize-barley orthogroups. Our analysis suggests that downregulation of GA-INSENSITIVE DWARF1 (GID1) and upregulation of α-Expansin1 (EXPA1) orthologs comprises a universal GA-response mechanism that is independent of GA biosynthesis, and identifies F-Box proteins, hexokinase, and AMPK/SNF1 protein kinase orthologs as unexpected cross-tissue, cross-genotype, and cross-species GA-responsive genes. We then compared the transient expression of the DR5, DR5v2, and DII-mDII auxin reporters in barley and maize and find that although DR5 did not respond to exogenous auxin in barley, DR5v2 responded to auxin treatment with a similar magnitude as in maize. Both species display auxin-mediated DII degradation that requires the 26S proteasome.
植物激素是引起深刻生理反应的小分子。虽然植物激素的生物合成和反应基因对农业改良至关重要,但由于复杂的表型输出,很难通过实验比较不同物种间的激素生物学。我们使用瞬时表达的基因激素传感器和转录组学来量化谷物作物组织和基因型间的组织特异性赤霉素(GA)和生长素反应。研究人员发现,基于fret的GPS2生物传感器可以在玉米、大麦、高粱和小麦的营养组织和花组织中检测外源GA处理。测量不同GA剂量的GPS2输出揭示了GA传感器反应的组织和基因型特异性差异。我们观察到玉米与大麦叶片和花组织的显著差异,以及玉米d1 GA生物合成突变体在GA处理后GPS2产量的意外下降,这可能反映了生物活性GA含量、GA运输和GA响应机制的差异。然后,我们使用RNAseq来测量玉米野生型、d1和大麦叶片以及玉米和大麦花组织对GA处理的转录反应,以进行跨组织、跨基因型和跨物种的GA反应比较。通过对种内和种间GO-term富集的同源预测和分析,我们确定了每个物种以及玉米-大麦同源群中ga响应基因的核心集合。我们的分析表明,GA不敏感的DWARF1 (GID1)的下调和α-扩张蛋白1 (EXPA1)同源物的上调构成了一种独立于GA生物合成的通用GA反应机制,并将F-Box蛋白、己糖激酶和AMPK/SNF1蛋白激酶同源物鉴定为意想不到的跨组织、跨基因型和跨物种的GA反应基因。然后,我们比较了DR5、DR5v2和DII-mDII生长素报告基因在大麦和玉米中的瞬时表达,发现尽管DR5在大麦中对外源生长素没有反应,但DR5v2对生长素处理的反应程度与玉米相似。这两个物种都表现出生长素介导的DII降解,这需要26S蛋白酶体。
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引用次数: 0
Dietary restriction and life-history trade-offs: insights into mTOR pathway regulation and reproductive investment in birds 饮食限制和生活史权衡:对鸟类mTOR通路调控和生殖投资的见解
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.14.567012
Gebrehaweria Kidane Reda, Sawadi Fransisco Ndunguru, Brigitta Csernus, Gabriella Gulyas, Renata Knop, Csaba Szabo, Levente Czegledi, Adam Z. Lendvai
Resources are needed for growth, reproduction and survival, and organisms must trade-off limited resources among competing processes. Nutritional availability in organisms is sensed and monitored by nutrient-sensing pathways that can trigger physiological changes or alter gene expression. Previous studies have proposed that one such signalling pathway, the mechanistic target of rapamycin (mTOR), underpins a form of adaptive plasticity when individuals encounter constraints in their energy budget. Despite the fundamental importance of this process in evolutionary biology, how nutritional limitation is regulated through the expression of genes governing this pathway and its consequential effects on fitness remains understudied, particularly in birds. We used dietary restriction to simulate resource depletion and examined its effects on body mass, reproduction and gene expression in Japanese quails (Coturnix japonica). Quails were subjected to ad libitum (ADL) feeding or 20%, 30%, and 40% restriction levels for two weeks. All restricted groups exhibited reduced body mass, whereas reductions in the number and mass of eggs were observed only under more severe restrictions. Dietary restriction led to decreased expression of mTOR and insulin-like growth factor 1 (IGF1), whereas the ribosomal protein S6 kinase 1 (RPS6K1) and autophagy-related genes (ATG9A and ATG5) were upregulated. The pattern in which mTOR respond to restriction was similar to what has been seen in body mass. Regardless of the treatment, proportionally higher reproductive investment was associated with individual variation in mTOR expression. These findings reveal the connection between dietary intake and the expression of mTOR and related genes in this pathway.
生长、繁殖和生存都需要资源,生物必须在竞争过程中权衡有限的资源。生物体内的营养可用性是通过营养感应途径感知和监测的,这些途径可以触发生理变化或改变基因表达。先前的研究已经提出,当个体在能量预算中遇到限制时,雷帕霉素(mTOR)的机制靶标(mechanistic target of rapamycin)这一信号通路支持一种形式的适应性可塑性。尽管这一过程在进化生物学中具有根本的重要性,但营养限制是如何通过控制这一途径的基因表达来调节的,以及它对适应性的影响仍未得到充分的研究,特别是在鸟类中。本研究以日本鹌鹑为研究对象,采用饮食限制模拟资源耗竭,研究其对日本鹌鹑体重、繁殖和基因表达的影响。鹌鹑分别饲喂20%、30%和40%限制水平的自由采食(ADL) 2周。所有受限制的组都表现出体重减少,而只有在更严格的限制下才观察到鸡蛋数量和质量的减少。饮食限制导致mTOR和胰岛素样生长因子1 (IGF1)表达降低,而核糖体蛋白S6激酶1 (RPS6K1)和自噬相关基因(ATG9A和ATG5)表达上调。mTOR对限制的反应模式与在体重中所看到的相似。无论治疗方式如何,比例较高的生殖投资与mTOR表达的个体差异相关。这些发现揭示了膳食摄入与mTOR及其相关基因表达之间的联系。
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引用次数: 0
Neopolyploidy-induced changes in the giant duckweed (Spirodela polyrhiza) alter herbivore preference, performance, and plant population performance 巨型浮萍(Spirodela polyrhiza)新多倍体诱导的变化改变了食草动物的偏好、性能和植物种群性能
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.14.567047
Hannah R Assour, Tia-Lynn Ashman, Martin M Turcotte
Premise: Polyploidy is a widespread mutational process in angiosperms that may alter population performance of not only plants but also their animal associates. Yet, knowledge of whether ploidy affects plant-herbivore dynamics is scarce. Here, we test whether aphid herbivores exhibit preference for diploid or neopolyploid plants, whether ploidy impacts plant and herbivore performance, and whether these interactions depend on plant genetic background. Methods: Using multiple pairs of independently synthesized neotetraploid greater duckweed (Spirodela polyrhiza) and their diploid progenitors, we evaluated the effect of neopolyploidy on duckweed's interaction with the water-lily aphid (Rhopalosiphum nymphaeae). Using two-way choice experiments, we first evaluated feeding preference by the herbivore. We then evaluated the consequences of ploidy on aphid and plant performance by measuring population growth over multiple generations. Key Results: Aphids preferred neopolyploids over diploids when the plants were provided at equal abundances but not when they were provided at equal surface area, indicating the role of plant size in driving this preference. Additionally, neopolyploidy increased aphid population performance, but this result was highly dependent on the genetic lineage of the plant. Lastly, the impact of herbivory on neopolyploids vs. diploid duckweed varied greatly with genetic lineage, but overall, neopolyploids appeared to be generally less tolerant than diploids. Conclusions: We conclude that polyploidization can impact the preference and performance of herbivores on their plant hosts, whereas plant performance depends on complex interactions between herbivory, ploidy, and genetic lineage. These results have significant implications for the establishment and persistence of plants and herbivores in nature.
前提:多倍体是被子植物中广泛存在的一种突变过程,它不仅可以改变植物的种群性能,也可以改变它们的动物同伴。然而,关于倍性是否影响植物-食草动物动力学的知识很少。在这里,我们测试了蚜虫食草动物是否表现出对二倍体或新多倍体植物的偏好,多倍体是否影响植物和食草动物的表现,以及这些相互作用是否依赖于植物遗传背景。方法:利用多对独立合成的新四倍体大浮萍(Spirodela polyrhiza)及其二倍体祖细胞,研究新多倍体对浮萍与睡菊蚜互作的影响。通过双向选择实验,我们首先评估了食草动物的摄食偏好。然后,我们通过测量多代的种群增长来评估倍性对蚜虫和植物性能的影响。关键结果:当植物丰度相等时,蚜虫对新多倍体的偏好高于二倍体,而当植物表面积相等时,蚜虫对新多倍体的偏好则不同,这表明植物大小在驱动这种偏好的作用。此外,新多倍体增加了蚜虫群体的表现,但这一结果高度依赖于植物的遗传谱系。最后,草食对新多倍体和二倍体浮萍的影响因遗传谱系而异,但总体而言,新多倍体的耐受性普遍低于二倍体。结论:多倍性可以影响食草动物对植物寄主的偏好和表现,而植物的表现取决于食草性、倍性和遗传谱系之间的复杂相互作用。这些结果对植物和食草动物在自然界的建立和持续具有重要意义。
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引用次数: 0
The Velvet transcription factor PnVeA regulates necrotrophic effectors and secondary metabolism in the wheat pathogenParastagonospora nodorum 丝绒转录因子PnVeA调控小麦病原菌芽孢对抗性菌的坏死效应物和次生代谢
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.13.566949
Shota Morikawa, Callum Verdonk, Evan John, Leon Lenzo, Nicolau Sbaraini, Chala J Turo, Hang Li, David Jiang, Yit-Heng Chooi, Kar-Chun Tan
The fungus Parastagonospora nodorum causes septoria nodorum blotch on wheat. The role of the fungal Velvet-family transcription factor VeA in P. nodorum development and virulence was investigated here. Deletion of the P. nodorum VeA ortholog, PnVeA , resulted in growth abnormalities including pigmentation, abolished asexual sporulation and highly reduced virulence on wheat. Comparative RNA-Seq and RT-PCR analyses revealed that the deletion of PnVeA also decoupled the expression of major necrotrophic effector genes. In addition, the deletion of PnVeA resulted in an up-regulation of four predicted secondary metabolite (SM) gene clusters. Using liquid-chromatography mass-spectrometry, it was observed that one of the SM gene clusters led to an accumulation of the mycotoxin alternariol. PnVeA is essential for asexual sporulation, full virulence, secondary metabolism and necrotrophic effector regulation.
真菌结核对抗性菌引起小麦上的结核斑病。本文研究了丝绒家族转录因子VeA在结核分枝杆菌(P. nodorum)发育和毒力中的作用。nodorum VeA同源基因PnVeA的缺失导致小麦的生长异常,包括色素沉着,无性孢子的减少和对小麦的毒力大大降低。比较RNA-Seq和RT-PCR分析显示,PnVeA的缺失也使主要的坏死性效应基因的表达分离。此外,PnVeA的缺失导致4个预测的次级代谢物(SM)基因簇的上调。利用液相色谱-质谱法,观察到其中一个SM基因簇导致真菌毒素互交霉醇的积累。PnVeA对无性孢子形成、全毒力、次生代谢和坏死性效应调节至关重要。
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引用次数: 0
Robust organ size in Arabidopsis is primarily governed by cell growth rather than cell division patterns 拟南芥健壮的器官大小主要由细胞生长而不是细胞分裂模式决定
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.11.566685
Isabella Burda, Chun-Biu Li, Frances K Clark, Adrienne HK Roeder
Organ sizes and shapes are highly reproducible, or robust, within a species and individuals. Arabidopsis thaliana sepals, which are the leaf-like organs that enclose flower buds, have consistent size and shape, which indicates robust development. Counterintuitively, variability in cell growth rate over time and between cells facilitates robust development because cumulative cell growth averages to a uniform rate. Here we investigate how sepal morphogenesis is robust to changes in cell division but not robust to changes in cell growth variability. We live image and quantitatively compare the development of sepals with increased or decreased cell division rate (lgo mutant and LGO overexpression, respectively), a mutant with altered cell growth variability (ftsh4), and double mutants combining these. We find that robustness is preserved when cell division rate changes because there is no change in the spatial pattern of growth. Meanwhile when robustness is lost in ftsh4 mutants, cell growth accumulates unevenly, and cells have disorganized growth directions. Thus, we demonstrate in vivo that both cell growth rate and direction average in robust development, preserving robustness despite changes in cell division.
器官的大小和形状在一个物种和个体中是高度可复制的。拟南芥萼片是包裹花蕾的叶状器官,其大小和形状一致,表明其发育健壮。与直觉相反的是,细胞生长速率随时间和细胞之间的变异性促进了健壮的发育,因为累积的细胞生长平均为统一的速率。在这里,我们研究萼片形态发生对细胞分裂的变化是稳健的,但对细胞生长变异性的变化不是稳健的。我们实时成像并定量比较了细胞分裂率增加或减少的萼片发育(分别为lgo突变体和lgo过表达)、细胞生长变异性改变的突变体(ftsh4)和结合这些突变体的双突变体。我们发现,当细胞分裂率发生变化时,由于生长的空间格局没有变化,因此鲁棒性得以保留。同时,当ftsh4突变体失去鲁棒性时,细胞生长积累不均匀,细胞生长方向混乱。因此,我们在体内证明了细胞生长速度和方向在稳健发育中都是平均的,尽管细胞分裂发生了变化,但仍保持稳健。
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引用次数: 0
Primary multistep phosphorelay activation comprises both cytokinin and abiotic stress responses in Brassicaceae 在十字花科植物中,主要的多步磷接力激活包括细胞分裂素和非生物胁迫响应
Pub Date : 2023-11-14 DOI: 10.1101/2023.11.14.567013
Katrina Leslie Nicolas Mala, Jan Skalak, Elena Zemlyanskaya, Vladislav Dolgikh, Veronika Jedlickova, Helene Robert-Boisivon, Lenka Havlickova, Klara Panzarova, Martin Trtilek, Ian Bancroft, Jan Hejatko
Multistep phosphorelay (MSP) signaling integrates hormonal and environmental signals to control plant development and adaptive responses. The type-A RESPONSE REGULATORs (RRAs), the downstream members of the MSP cascade and cytokinin primary response genes, are supposed to mediate primarily the negative feedback regulation of (cytokinin-induced) MSP signaling. However, the transcriptional data suggest the involvement of RRAs in stress-related responses as well. By employing evolutionary conservation with the well-characterized Arabidopsis thaliana RRAs, we identified 5 and 38 novel putative RRAs in Brassica oleracea and Brassica napus, respectively. Our phylogenetic analysis suggests the existence of gene-specific selective pressure, maintaining the homologs of ARR3, ARR6, and ARR16 as singletons during the evolution of Brassica oleracea and Brassica rapa. We categorized RRAs based on the kinetics of their cytokinin-mediated upregulation and observed both similarities and specificities in this type of response across Brassicaceae. Using bioinformatic analysis and experimental data demonstrating the cytokinin responsiveness of Arabidopsis-derived TCSv2 reporter, we unveil the mechanistic conservation of cytokinin-mediated upregulation of RRAs in Brassica rapa and Brassica napus. Notably, we identify partial cytokinin dependency of cold stress-induced RRA transcription, thus corroborating the role of cytokinin signaling in the crop adaptive responses.
多步磷接力(Multistep phosphorelay, MSP)信号通过整合激素和环境信号来控制植物的发育和适应性反应。a型反应调节因子(type-A RESPONSE REGULATORs, RRAs)是MSP级联的下游成员和细胞分裂素一级反应基因,被认为主要介导(细胞分裂素诱导的)MSP信号的负反馈调节。然而,转录数据表明RRAs也参与应激相关反应。通过对拟南芥RRAs的进化保守分析,我们在甘蓝和甘蓝型油菜中分别鉴定出5个和38个新的推测RRAs。我们的系统发育分析表明,在甘蓝和油菜的进化过程中,存在基因特异性的选择压力,使ARR3、ARR6和ARR16的同源基因保持为单基因。我们根据细胞分裂素介导的上调动力学对RRAs进行了分类,并观察了十字花科植物中这种反应的相似性和特异性。利用生物信息学分析和实验数据证明了拟南芥来源的TCSv2报告细胞分裂素的响应性,我们揭示了细胞分裂素介导的RRAs上调在油菜和甘蓝型油菜中的机制保护。值得注意的是,我们发现冷胁迫诱导的RRA转录部分依赖细胞分裂素,从而证实了细胞分裂素信号在作物适应性反应中的作用。
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引用次数: 0
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bioRxiv (Cold Spring Harbor Laboratory)
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