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Comprehensive comparative analysis and development of molecular markers for Lasianthus species based on complete chloroplast genome sequences 基于完整叶绿体基因组序列的 Lasianthus 品种分子标记的综合比较分析与开发
IF 5.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-12-31 DOI: 10.1186/s12870-024-05383-z
Yue Zhang, Meifang Song, Deying Tang, Xianjing Li, Niaojiao Xu, Haitao Li, Lu Qu, Yunqiang Wang, Cuiyun Yin, Lixia Zhang, Zhonglian Zhang
Lasianthus species are widely used in traditional Chinese folk medicine with high medicinal value. However, source materials and herbarium specimens are often misidentified due to morphological characteristics and commonly used DNA barcode fragments are not sufficient for accurately identifying Lasianthus species. To improve the molecular methods for distinguishing among Lasianthus species, we report the complete chloroplast (CP) genomes of Lasianthus attenuatus, Lasianthus henryi, Lasianthus hookeri, Lasianthus sikkimensis, obtained via high-throughput Illumina sequencing. These showed CP genomes size of 160164-160246 bp and a typical quadripartite structure, including a large single-copy region (86675–86848 bp), a small single-copy region (17177–17326 bp), and a pair of inverted repeats (28089–28135 bp). As a whole, the gene order, GC content and IR/SC boundary structure were remarkably similar among of the four Lasianthus CP genomes, the partial gene length and IR, LSC and SSC regions length are still different. The average GC content of the CP genomes was 36.71–36.75%, and a total of 129 genes were detected, including 83 different protein-coding genes, 8 different rRNA genes and 38 different tRNA genes. Furthermore, we compared our 4 complete CP genomes data with publicly available CP genome data from six other Lasianthus species, and we initially screened eleven highly variable region fragments were initially screened. We then evaluated the identification efficiency of eleven highly variable region fragments and 5 regular barcode fragments. Ultimately, we found that the optimal combination fragment' ITS2 + psaI-ycf4' could authenticated the Lasianthus species well. Additionally, the results of genome comparison of Rubiaceae species showed that the coding region is more conservative than the non-coding region, and the ycf1 gene shows the most significant variation. Finally, 49 species of CP genome sequences belonging to 16 genera of the Rubiaceae family were used to construct phylogenetic trees. Our research is the first to analyze the chloroplast genomes of four species of Lasianthus in detail and we ultimately determined that the combination fragment' ITS2 + psaI-ycf4' is the optimal barcode combination for identifying the genus of Lasianthus. Meanwhile, we gathered the available CP genome sequences from the Rubiaceae and used them to construct the most comprehensive phylogenetic tree for the Rubiaceae family. These investigations provide an important reference point for further studies in the species identification, genetic diversity, and phylogenetic analyses of Rubiaceae species.
桔梗被广泛用于传统中药中,具有很高的药用价值。然而,由于形态特征的原因,原始材料和标本馆标本经常被误认,而常用的 DNA 条形码片段也不足以准确鉴定桔梗属植物。为了改进分子鉴定方法,我们报告了通过高通量 Illumina 测序获得的 Lasianthus attenuatus、Lasianthus henryi、Lasianthus hookeri 和 Lasianthus sikkimensis 的完整叶绿体(CP)基因组。它们的 CP 基因组大小为 160164-160246 bp,具有典型的四方结构,包括一个大的单拷贝区(86675-86848 bp)、一个小的单拷贝区(17177-17326 bp)和一对倒位重复区(28089-28135 bp)。从整体上看,4个Lasianthus CP基因组的基因顺序、GC含量和IR/SC边界结构非常相似,但部分基因长度和IR、LSC和SSC区长度仍有差异。CP基因组的平均GC含量为36.71%-36.75%,共检测到129个基因,包括83个不同的蛋白编码基因、8个不同的rRNA基因和38个不同的tRNA基因。此外,我们将 4 个完整的 CP 基因组数据与其他 6 个 Lasianthus 品种的 CP 基因组数据进行了比较,初步筛选出 11 个高变异区片段。然后,我们评估了 11 个高度可变区片段和 5 个常规条形码片段的鉴定效率。最终,我们发现最佳组合片段'ITS2 + psaI-ycf4'能很好地鉴定腊梅属植物。此外,茜草科植物基因组比较结果表明,编码区比非编码区保守,其中 ycf1 基因的变异最为显著。最后,我们利用茜草科 16 属 49 个物种的 CP 基因组序列构建了系统发生树。我们的研究首次详细分析了四种茜草属植物的叶绿体基因组,并最终确定'ITS2 + psaI-ycf4'组合片段是鉴定茜草属植物的最佳条形码组合。同时,我们收集了茜草科现有的 CP 基因组序列,并利用这些序列构建了最全面的茜草科系统发生树。这些研究为进一步研究茜草科植物的物种鉴定、遗传多样性和系统发育分析提供了重要参考。
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引用次数: 0
Deciphering the complex organelle genomes of two Rhododendron species and insights into adaptive evolution patterns in high-altitude. 解密两个杜鹃花物种的复杂细胞器基因组,洞察高海拔地区的适应性进化模式。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-08 DOI: 10.1186/s12870-024-05761-7
Zhen-Yu Lyu, Gao-Ming Yang, Xiong-Li Zhou, Si-Qi Wang, Rui Zhang, Shi-Kang Shen

Background: The genomes within organelles are crucial for physiological functions such as respiration and photosynthesis and may also contribute to environmental adaptation. However, the limited availability of genetic resources, particularly mitochondrial genomes, poses significant challenges for in-depth investigations.

Results: Here, we explored various assembly methodologies and successfully reconstructed the complex organelle genomes of two Rhododendron species: Rhododendron nivale subsp. boreale and Rhododendron vialii. The mitogenomes of these species exhibit various conformations, as evidenced by long-reads mapping. Notably, only the mitogenome of R. vialii can be depicted as a singular circular molecule. The plastomes of both species conform to the typical quadripartite structure but exhibit elongated inverted repeat (IR) regions. Compared to the high similarity between plastomes, the mitogenomes display more obvious differences in structure, repeat sequences, and codon usage. Based on the analysis of 58 organelle genomes from angiosperms inhabiting various altitudes, we inferred the genetic adaptations associated with high-altitude environments. Phylogenetic analysis revealed partial inconsistencies between plastome- and mitogenome-derived phylogenies. Additionally, evolutionary lineage was determined to exert a greater influence on codon usage than altitude. Importantly, genes such as atp4, atp9, mttB, and clpP exhibited signs of positive selection in several high-altitude species, suggesting a potential link to alpine adaptation.

Conclusions: We tested the effectiveness of different organelle assembly methods for dealing with complex genomes, while also providing and validating high-quality organelle genomes of two Rhododendron species. Additionally, we hypothesized potential strategies for high-altitude adaptation of organelles. These findings offer a reference for the assembly of complex organelle genomes, while also providing new insights and valuable resources for understanding their adaptive evolution patterns.

背景:细胞器内的基因组对呼吸和光合作用等生理功能至关重要,也可能有助于环境适应。然而,有限的遗传资源,尤其是线粒体基因组,给深入研究带来了巨大挑战:在此,我们探索了各种组装方法,并成功重建了两个杜鹃花物种的复杂细胞器基因组:boreale 和 Rhododendron vialii。这些物种的有丝分裂基因组表现出不同的构象,长线程图谱就是证明。值得注意的是,只有 R. vialii 的有丝分裂原体可以被描绘成一个单一的环状分子。这两个物种的质粒都符合典型的四方结构,但表现出拉长的倒置重复(IR)区域。与质粒之间的高度相似性相比,有丝分裂原质粒在结构、重复序列和密码子使用方面的差异更为明显。基于对生活在不同海拔地区的被子植物的 58 个细胞器基因组的分析,我们推断了与高海拔环境相关的遗传适应性。系统发育分析表明,质粒体和有丝分裂原衍生的系统发育之间存在部分不一致。此外,与海拔相比,进化系对密码子使用的影响更大。重要的是,在几个高海拔物种中,atp4、atp9、mttB 和 clpP 等基因表现出正选择的迹象,这表明它们与高山适应有潜在联系:我们测试了处理复杂基因组的不同细胞器组装方法的有效性,同时还提供并验证了两个杜鹃花物种的高质量细胞器基因组。此外,我们还假设了细胞器高海拔适应的潜在策略。这些发现为组装复杂细胞器基因组提供了参考,同时也为了解其适应性进化模式提供了新的见解和宝贵的资源。
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引用次数: 0
Promoting tomato resilience: effects of ascorbic acid and sulfur-treated biochar in saline and non-saline cultivation environments. 提高番茄的抗逆性:抗坏血酸和硫处理生物炭在盐碱和非盐碱栽培环境中的效果。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-08 DOI: 10.1186/s12870-024-05734-w
Muhammad Ikram, Asif Minhas, Arwa A Al-Huqail, Adel M Ghoneim, Sammina Mahmood, Esawy Mahmoud, Maryam Tahira, Muhammad Mehran, Muhammad Faizan Khurram Maqsood, Abdul Rauf, Waqar Ali

The resilience of tomato plants under different cultivation environments, particularly saline and non-saline conditions, was investigated by applying various treatments, including 0.5% Ascorbic Acid (AsA) and 1% Sulphur-treated Biochar (BS). The study evaluated parameters such as fruit length, diameter, yield per plant and pot, Total Soluble Solids (TSS) content, chlorophyll content, electrolyte leakage, enzyme activities (Superoxide Dismutase - SOD, Peroxidase - POD, Catalase - CAT), and nutrient content (Nitrogen - N%, Phosphorus - P%, Potassium - K%). Under saline conditions, significant enhancements were observed in fruit characteristics and yield metrics with the application of AsA and BS individually, with the combined treatment yielding the most substantial improvements. Notably, AsA and BS treatments exhibited varying effects on TSS levels, chlorophyll content, electrolyte leakage, and enzyme activities, with the combination treatment consistently demonstrating superior outcomes. Additionally, nutrient content analysis revealed notable increases, particularly under non-saline conditions, with the combined treatment showcasing the most significant enhancements. Overall, the study underscores the potential of AsA and BS treatments in promoting tomato resilience, offering insights into their synergistic effects on multiple physiological and biochemical parameters crucial for plant growth and productivity.

通过采用各种处理方法,包括 0.5% 抗坏血酸 (AsA) 和 1% 硫处理生物炭 (BS),研究了番茄植株在不同栽培环境,特别是盐碱和非盐碱条件下的抗逆性。研究评估了果实长度、直径、每株和每盆产量、总可溶性固形物(TSS)含量、叶绿素含量、电解质渗漏、酶活性(超氧化物歧化酶 - SOD、过氧化物酶 - POD、过氧化氢酶 - CAT)和养分含量(氮 - N%、磷 - P%、钾 - K%)等参数。在盐碱条件下,单独施用 AsA 和 BS 能显著改善果实特征和产量指标,而联合施用则能获得最大改善。值得注意的是,AsA 和 BS 处理对总悬浮固体含量、叶绿素含量、电解质渗漏和酶活性的影响各不相同,而组合处理的效果始终更佳。此外,营养成分分析表明,特别是在非盐水条件下,营养成分含量明显增加,其中组合处理的改善效果最为显著。总之,该研究强调了 AsA 和 BS 处理在促进番茄抗逆性方面的潜力,并深入探讨了它们对植物生长和生产力至关重要的多个生理和生化参数的协同作用。
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引用次数: 0
Assembly and comparative analysis of the first complete mitochondrial genome of Astragalus membranaceus (Fisch.) Bunge: an invaluable traditional Chinese medicine. 黄芪(Fisch.)
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-08 DOI: 10.1186/s12870-024-05780-4
Kun Zhang, Gaoyang Qu, Yue Zhang, Jianxia Liu
<p><strong>Background: </strong>Astragalus membranaceus (Fisch.) Bunge is one of the most well-known tonic herbs in traditional Chinese medicine, renowned for its remarkable medicinal value in various clinical contexts. The corresponding chloroplast (cp) and nuclear genomes have since been accordingly sequenced, providing valuable information for breeding and phylogeny studies. However, the mitochondrial genome (mitogenome) of A. membranaceus remains unexplored, which hinders comprehensively understanding the evolution of its genome.</p><p><strong>Results: </strong>For this study, we de novo assembled the mitogenome of A. membranaceus (Fisch.) Bunge var. mongholicus (Bunge) P. K. Hsiao using a strategy integrating Illumina and Nanopore sequencing technology and subsequently performed comparative analysis with its close relatives. The mitogenome has a multi-chromosome structure, consisting of two circular chromosomes with a total length of 398,048 bp and an overall GC content of 45.3%. It encodes 54 annotated functional genes, comprising 33 protein-coding genes (PCGs), 18 tRNA genes, and 3 rRNA genes. An investigation of codon usage in the PCGs revealed an obvious preference for codons ending in A or U (T) bases, given their high frequency. RNA editing identified 500 sites in the coding regions of mt PCGs that exhibit a perfect conversion of the base C to U, a process that tends to lead to the conversion of hydrophilic amino acids into hydrophobic amino acids. From the mitogenome analysis, a total of 399 SSRs, 4 tandem repeats, and 77 dispersed repeats were found, indicating that A. membranaceus possesses fewer repeats compared to its close relatives with similarly sized mitogenomes. Selection pressure analysis indicated that most mt PCGs were purifying selection genes, while only five PCGs (ccmB, ccmFc, ccmFn, nad3, and nad9) were positive selection genes. Notably, positive selection emerged as a critical factor in the evolution of ccmB and nad9 in all the pairwise species comparisons, suggesting the extremely critical role of these genes in the evolution of A. membranaceus. Moreover, we inferred that 22 homologous fragments have been transferred from cp to mitochondria (mt), in which 5 cp-derived tRNA genes remain intact in the mitogenome. Further comparative analysis revealed that the syntenic region and mt gene organization are relatively conserved within the provided legumes. The comparison of gene content indicated that the gene composition of Fabaceae mitogenomes differed. Finally, the phylogenetic tree established from analysis is largely congruent with the taxonomic relationships of Fabaceae species and highlights the close relationship between Astragalus and Oxytropis.</p><p><strong>Conclusions: </strong>We provide the first report of the assembled and annotated A. membranaceus mitogenome, which enriches the genetic resources available for the Astragalus genus and lays the foundation for comprehensive exploration of this invaluable medic
背景:黄芪(Astragalus membranaceus (Fisch.) Bunge)是传统中药中最著名的滋补药材之一,因其在各种临床情况下的显著药用价值而闻名。相应的叶绿体(cp)基因组和核基因组已经完成测序,为育种和系统发育研究提供了宝贵的信息。然而,A. membranaceus 的线粒体基因组(有丝分裂基因组)仍未被探索,这阻碍了对其基因组进化的全面了解:结果:在本研究中,我们利用 Illumina 和 Nanopore 测序技术,从头组装了膜草属植物(A. membranaceus (Fisch.) Bunge var.有丝分裂基因组具有多染色体结构,由两条环状染色体组成,总长度为 398,048 bp,总 GC 含量为 45.3%。它编码 54 个注释功能基因,包括 33 个蛋白质编码基因(PCGs)、18 个 tRNA 基因和 3 个 rRNA 基因。对 PCGs 中密码子使用情况的调查显示,由于以 A 或 U(T)碱基结尾的密码子使用频率较高,因此这些密码子明显更受青睐。RNA 编辑发现,在 mt PCGs 的编码区中有 500 个位点显示出碱基 C 到 U 的完美转换,这一过程往往会导致亲水性氨基酸转换为疏水性氨基酸。在有丝分裂基因组分析中,共发现了 399 个 SSR、4 个串联重复序列和 77 个分散重复序列,这表明与有丝分裂基因组大小相似的近缘种相比,膜衣壳属植物的重复序列较少。选择压力分析表明,大多数 mt PCGs 是纯化选择基因,只有 5 个 PCGs(ccmB、ccmFc、ccmFn、nad3 和 nad9)是正向选择基因。值得注意的是,在所有物种配对比较中,正选择都是ccmB和nad9进化的关键因素,这表明这些基因在膜草属植物的进化中起着极其关键的作用。此外,我们还推断出 22 个同源片段已从 cp 转移到线粒体(mt),其中 5 个 cp 衍生的 tRNA 基因在有丝分裂基因组中保持完整。进一步的比较分析表明,在所提供的豆科植物中,同源区和 mt 基因组织相对保守。基因含量的比较表明,豆科植物有丝分裂基因组的基因组成存在差异。最后,通过分析建立的系统发生树与豆科植物的分类关系基本一致,并突出了黄芪与牛膝草之间的密切关系:我们首次报道了组装和注释的黄芪有丝分裂基因组,这丰富了黄芪属的遗传资源,为全面探索这一珍贵的药用植物奠定了基础。
{"title":"Assembly and comparative analysis of the first complete mitochondrial genome of Astragalus membranaceus (Fisch.) Bunge: an invaluable traditional Chinese medicine.","authors":"Kun Zhang, Gaoyang Qu, Yue Zhang, Jianxia Liu","doi":"10.1186/s12870-024-05780-4","DOIUrl":"https://doi.org/10.1186/s12870-024-05780-4","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Astragalus membranaceus (Fisch.) Bunge is one of the most well-known tonic herbs in traditional Chinese medicine, renowned for its remarkable medicinal value in various clinical contexts. The corresponding chloroplast (cp) and nuclear genomes have since been accordingly sequenced, providing valuable information for breeding and phylogeny studies. However, the mitochondrial genome (mitogenome) of A. membranaceus remains unexplored, which hinders comprehensively understanding the evolution of its genome.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;For this study, we de novo assembled the mitogenome of A. membranaceus (Fisch.) Bunge var. mongholicus (Bunge) P. K. Hsiao using a strategy integrating Illumina and Nanopore sequencing technology and subsequently performed comparative analysis with its close relatives. The mitogenome has a multi-chromosome structure, consisting of two circular chromosomes with a total length of 398,048 bp and an overall GC content of 45.3%. It encodes 54 annotated functional genes, comprising 33 protein-coding genes (PCGs), 18 tRNA genes, and 3 rRNA genes. An investigation of codon usage in the PCGs revealed an obvious preference for codons ending in A or U (T) bases, given their high frequency. RNA editing identified 500 sites in the coding regions of mt PCGs that exhibit a perfect conversion of the base C to U, a process that tends to lead to the conversion of hydrophilic amino acids into hydrophobic amino acids. From the mitogenome analysis, a total of 399 SSRs, 4 tandem repeats, and 77 dispersed repeats were found, indicating that A. membranaceus possesses fewer repeats compared to its close relatives with similarly sized mitogenomes. Selection pressure analysis indicated that most mt PCGs were purifying selection genes, while only five PCGs (ccmB, ccmFc, ccmFn, nad3, and nad9) were positive selection genes. Notably, positive selection emerged as a critical factor in the evolution of ccmB and nad9 in all the pairwise species comparisons, suggesting the extremely critical role of these genes in the evolution of A. membranaceus. Moreover, we inferred that 22 homologous fragments have been transferred from cp to mitochondria (mt), in which 5 cp-derived tRNA genes remain intact in the mitogenome. Further comparative analysis revealed that the syntenic region and mt gene organization are relatively conserved within the provided legumes. The comparison of gene content indicated that the gene composition of Fabaceae mitogenomes differed. Finally, the phylogenetic tree established from analysis is largely congruent with the taxonomic relationships of Fabaceae species and highlights the close relationship between Astragalus and Oxytropis.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;We provide the first report of the assembled and annotated A. membranaceus mitogenome, which enriches the genetic resources available for the Astragalus genus and lays the foundation for comprehensive exploration of this invaluable medic","PeriodicalId":9198,"journal":{"name":"BMC Plant Biology","volume":null,"pages":null},"PeriodicalIF":4.3,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The role of gibberellin synthase gene VvGA2ox7 acts as a positive regulator to salt stress in Arabidopsis thaliana. 拟南芥赤霉素合成酶基因 VvGA2ox7 对盐胁迫的正向调节作用
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-07 DOI: 10.1186/s12870-024-05708-y
Huimin Gou, Shixiong Lu, Guojie Nai, Weifeng Ma, Jiaxuan Ren, Lili Guo, Baihong Chen, Juan Mao

Background: Soil salinity is an important environmental component affecting plant growth and yield, but high-salinity soils are a major constraint to the development of the grape industry. Previous studies have provided lines of evidence that gibberellins (GAs) play a significant regulatory role in plant responses to salt stress. However, it remains unclear whether GA2ox, a key enzyme that maintains the balance of bioactive gibberellins and intermediates in plants, is involved in the mechanism of salt stress tolerance in grapes.

Results: In this study, we found that GA2ox7 positively modulates salt stress via its ectopic expression in Arabidopsis thaliana. The GA2ox7 gene cloned from grape was a hydrophilic protein, its CDS length was 1002 bp. Besides, VvGA2ox7 protein contained DIOX_N and 2OG-FeII_Oxy domains and was localized at the nucleus and cytoplasm. Yeast two-hybrid (Y2H) showed VvARCN1, VvB5R, VvRUB2, and VvCAR11 might be potential interaction proteins of VvGA2ox7. Compared with the wild type, overexpression of VvGA2ox7 in Arabidopsis thaliana enhanced antioxidant enzymatic activities and proline, chlorophyll, and ABA contents, and decreased relative electrical conductivity, malondialdehyde, and GA3 contents. Moreover, overexpression of VvGA2ox7 positively regulated the expression of salt stress response genes (KAT1, APX1, LEA, P5CS1, AVP1, CBF1), indicating that the VvGA2ox7 overexpression improved the salt stress tolerance of plants.

Conclusions: Taken together, this investigation indicates that VvGA2ox7 may act as a positive regulator in response to salt stress and provide novel insights for a deeper understanding of the role of VvGA2ox7 in grapes.

背景:土壤盐分是影响植物生长和产量的重要环境因素,但高盐分土壤是葡萄产业发展的主要制约因素。以前的研究已经提供了一系列证据,证明赤霉素(GA)在植物对盐胁迫的反应中起着重要的调节作用。然而,GA2ox(一种维持植物体内生物活性赤霉素和中间产物平衡的关键酶)是否参与了葡萄耐盐胁迫的机制,目前仍不清楚:结果:在这项研究中,我们发现 GA2ox7 通过在拟南芥中异位表达对盐胁迫有积极的调节作用。从葡萄中克隆的GA2ox7基因是一种亲水蛋白,其CDS长度为1002 bp。此外,VvGA2ox7蛋白含有DIOX_N和2OG-FeII_Oxy结构域,定位于细胞核和细胞质。酵母双杂交(Y2H)显示 VvARCN1、VvB5R、VvRUB2 和 VvCAR11 可能是 VvGA2ox7 潜在的相互作用蛋白。与野生型相比,VvGA2ox7在拟南芥中的过表达提高了抗氧化酶活性、脯氨酸、叶绿素和ABA含量,降低了相对电导率、丙二醛和GA3含量。此外,VvGA2ox7的过表达还能正向调节盐胁迫响应基因(KAT1、APX1、LEA、P5CS1、AVP1、CBF1)的表达,表明VvGA2ox7的过表达能提高植物对盐胁迫的耐受性:综上所述,这项研究表明 VvGA2ox7 可能是应对盐胁迫的正调控因子,为深入了解 VvGA2ox7 在葡萄中的作用提供了新的见解。
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引用次数: 0
Establishment and application of highly efficient regeneration, genetic transformation and genome editing system for cucurbitacins biosynthesis in Hemsleya chinensis. 用于葫芦素生物合成的高效再生、遗传转化和基因组编辑系统的建立与应用。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-07 DOI: 10.1186/s12870-024-05717-x
Juan Wang, Chao-Hui Li, Chun-Fan Xiang, Pin-Han Zhou, Le-Song Li, Xia Li, Sheng-Chao Yang, Guang-Hui Zhang, Yan Zhao

Background: Hemsleya Chinensis is a perennial plant in the Cucurbitaceae family containing antibacterial and anti-inflammatory compounds. The lack of genetic transformation systems makes it difficult to verify the functions of genes controlling important traits and conduct molecular breeding in H. chinensis.

Results: Highly efficient calli were induced on MS medium added 1.5 mg·L- 1 6-benzylaminopurine (6-BA) and 0.02 mg·L- 1 1-naphthylacetic acid (NAA) with high efficiency (> 95%). The frequency of shoot induction was increased to 90% with a plant growth regulator combination of 1.5 mg·L- 1 6-BA and 0.1 mg·L- 1 NAA. Our results also showed that 100% of shoot regeneration was achieved in a shoot regeneration medium. Additionally, more than 92% of kanamycin-resistant plants were confirmed. Furthermore, we achieved 42% genome editing efficiency by applying this transformation method to HcOSC6, a gene that catalyzes the formation of cucurbitadienol. HPLC analysis showed OE-HcOSC6 lines exhibited significantly higher cucurbitadienol levels than the genome-edited lines. Transcriptomic analysis revealed that some downstream genes related to cucurbitadienol biosynthesis, such as HcCYP87D20, HcCYP81Q58, and HcSDR34, were up-regulated in OE lines and down-regulated in mutants.

Conclusions: Here, we established a process for regeneration, transformation, and genome editing of H. chinensis using stem segments. This provides valuable insight into the underlying molecular mechanisms of medicinal compound production. By combining high-efficiency tissue culture, transformation, and genome editing systems, we provide a powerful platform that supports functional research on molecular mechanisms of secondary metabolism.

背景:鹤望兰(Hemsleya Chinensis)是葫芦科多年生植物,含有抗菌和消炎化合物。由于缺乏遗传转化系统,很难验证控制其重要性状的基因的功能,也很难进行分子育种:结果:在添加了 1.5 mg-L- 1 6-苄基氨基嘌呤(6-BA)和 0.02 mg-L- 1 1-萘乙酸(NAA)的 MS 培养基上诱导出高效胼胝体,诱导效率高(> 95%)。使用 1.5 mg-L- 1 6-BA 和 0.1 mg-L- 1 NAA 的植物生长调节剂组合后,新芽诱导率提高到 90%。我们的结果还显示,在芽再生培养基中,芽的再生率达到了 100%。此外,超过 92% 的抗卡那霉素植株得到了确认。此外,通过对催化葫芦二烯醇形成的基因 HcOSC6 应用这种转化方法,我们实现了 42% 的基因组编辑效率。高效液相色谱分析显示,OE-HcOSC6 株系的葫芦烯醇含量明显高于基因组编辑株系。转录组分析表明,与葫芦二烯醇生物合成相关的一些下游基因,如 HcCYP87D20、HcCYP81Q58 和 HcSDR34,在 OE 株系中上调,而在突变体中下调:结论:在此,我们建立了一种利用茎段再生、转化和基因组编辑 H. chinensis 的方法。结论:在此,我们建立了利用茎段进行再生、转化和基因组编辑的过程,这为深入了解药用化合物生产的分子机制提供了宝贵的视角。通过结合高效的组织培养、转化和基因组编辑系统,我们提供了一个强大的平台,支持对次生代谢分子机制的功能研究。
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引用次数: 0
Exploring the response of yellow lupine (Lupinus luteus L.) root to drought mediated by pathways related to phytohormones, lipid, and redox homeostasis. 探索植物激素、脂质和氧化还原平衡相关途径介导的黄羽扇豆(Lupinus luteus L.)根对干旱的响应。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-06 DOI: 10.1186/s12870-024-05748-4
Sebastian Burchardt, Małgorzata Czernicka, Agata Kućko, Wojciech Pokora, Małgorzata Kapusta, Krzysztof Domagalski, Katarzyna Jasieniecka-Gazarkiewicz, Jacek Karwaszewski, Emilia Wilmowicz

Background: Yellow lupine (Lupinus luteus L.) is a high-protein crop of considerable economic and ecological significance. It has the ability to fix atmospheric nitrogen in symbiosis with Rhizobium, enriching marginal soils with this essential nutrient and reducing the need for artificial fertilizers. Additionally, lupine produces seeds with a high protein content, making it valuable for animal feed production. However, drought negatively affects lupine development, its mutualistic relationship with bacteria, and overall yield. To understand how lupine responds to this stress, global transcriptome sequencing was conducted, along with in-depth biochemical, chromatography, and microscopy analyses of roots subjected to drought. The results presented here contribute to strategies aimed at mitigating the effects of water deficit on lupine growth and development.

Results: Based on RNA-seq, drought-specific genes were identified and annotated to biological pathways involved in phytohormone biosynthesis/signaling, lipid metabolism, and redox homeostasis. Our findings indicate that drought-induced disruption of redox balance characterized by the upregulation of reactive oxygen species (ROS) scavenging enzymes, coincided with the accumulation of lipid-metabolizing enzymes, such as phospholipase D (PLD) and lipoxygenase (LOX). This disruption also led to modifications in lipid homeostasis, including increased levels of triacylglycerols (TAG) and free fatty acids (FFA), along with a decrease in polar lipid content. Additionally, the stress response involved alterations in the transcriptional regulation of the linolenic acid metabolism network, resulting in changes in the composition of fatty acids containing 18 carbons.

Conclusion: The first comprehensive global transcriptomic profiles of lupine roots, combined with the identification of key stress-responsive molecules, represent a significant advancement in understanding lupine's responses to abiotic stress. The increased expression of the Δ12DESATURASE gene and enhanced PLD activity lead to higher level of linoleic acid (18:2), which is subsequently oxidized by LOX, resulting in membrane damage and malondialdehyde (MDA) accumulation. Oxidative stress elevates the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), while the conversion of FFAs into TAGs provides protection against ROS. This research offers valuable molecular and biochemical candidates with significant potential to enhance drought tolerance . It enables innovative strategies in lupine breeding and crop improvement to address critical agricultural challenges.

背景:黄羽扇豆(Lupinus luteus L.)是一种高蛋白作物,具有重要的经济和生态意义。它能与根瘤菌共生固定大气中的氮,使贫瘠的土壤富含这种必需的养分,减少对人工肥料的需求。此外,羽扇豆生产的种子蛋白质含量高,因此对动物饲料生产很有价值。然而,干旱会对羽扇豆的生长发育、它与细菌的互助关系以及总体产量产生负面影响。为了了解羽扇豆是如何应对这种压力的,研究人员进行了全球转录组测序,并对遭受干旱的根部进行了深入的生化、色谱和显微分析。本文介绍的结果有助于制定旨在减轻缺水对羽扇豆生长和发育影响的策略:结果:基于RNA-seq分析,发现了干旱特异性基因,并将其注释到涉及植物激素生物合成/信号传导、脂质代谢和氧化还原平衡的生物通路中。我们的研究结果表明,干旱诱导的氧化还原平衡破坏以活性氧(ROS)清除酶的上调为特征,与磷脂酶 D(PLD)和脂氧酶(LOX)等脂质代谢酶的积累同时发生。这种破坏也导致了脂质平衡的改变,包括三酰甘油(TAG)和游离脂肪酸(FFA)含量的增加,以及极性脂质含量的减少。此外,应激反应还涉及亚麻酸代谢网络转录调控的改变,导致含有 18 个碳原子的脂肪酸组成发生变化:羽扇豆根部首次全面的全球转录组图谱以及关键胁迫响应分子的鉴定,是了解羽扇豆对非生物胁迫响应的重大进展。Δ12DESATURASE基因表达的增加和PLD活性的增强导致亚油酸(18:2)水平升高,亚油酸随后被LOX氧化,造成膜损伤和丙二醛(MDA)积累。氧化应激会提高超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)和过氧化氢酶(CAT)的活性,而将 FFAs 转化为 TAGs 则提供了抵御 ROS 的保护。这项研究提供了宝贵的分子和生化候选物质,具有提高耐旱性的巨大潜力。它为羽扇豆育种和作物改良提供了创新战略,以应对严峻的农业挑战。
{"title":"Exploring the response of yellow lupine (Lupinus luteus L.) root to drought mediated by pathways related to phytohormones, lipid, and redox homeostasis.","authors":"Sebastian Burchardt, Małgorzata Czernicka, Agata Kućko, Wojciech Pokora, Małgorzata Kapusta, Krzysztof Domagalski, Katarzyna Jasieniecka-Gazarkiewicz, Jacek Karwaszewski, Emilia Wilmowicz","doi":"10.1186/s12870-024-05748-4","DOIUrl":"10.1186/s12870-024-05748-4","url":null,"abstract":"<p><strong>Background: </strong>Yellow lupine (Lupinus luteus L.) is a high-protein crop of considerable economic and ecological significance. It has the ability to fix atmospheric nitrogen in symbiosis with Rhizobium, enriching marginal soils with this essential nutrient and reducing the need for artificial fertilizers. Additionally, lupine produces seeds with a high protein content, making it valuable for animal feed production. However, drought negatively affects lupine development, its mutualistic relationship with bacteria, and overall yield. To understand how lupine responds to this stress, global transcriptome sequencing was conducted, along with in-depth biochemical, chromatography, and microscopy analyses of roots subjected to drought. The results presented here contribute to strategies aimed at mitigating the effects of water deficit on lupine growth and development.</p><p><strong>Results: </strong>Based on RNA-seq, drought-specific genes were identified and annotated to biological pathways involved in phytohormone biosynthesis/signaling, lipid metabolism, and redox homeostasis. Our findings indicate that drought-induced disruption of redox balance characterized by the upregulation of reactive oxygen species (ROS) scavenging enzymes, coincided with the accumulation of lipid-metabolizing enzymes, such as phospholipase D (PLD) and lipoxygenase (LOX). This disruption also led to modifications in lipid homeostasis, including increased levels of triacylglycerols (TAG) and free fatty acids (FFA), along with a decrease in polar lipid content. Additionally, the stress response involved alterations in the transcriptional regulation of the linolenic acid metabolism network, resulting in changes in the composition of fatty acids containing 18 carbons.</p><p><strong>Conclusion: </strong>The first comprehensive global transcriptomic profiles of lupine roots, combined with the identification of key stress-responsive molecules, represent a significant advancement in understanding lupine's responses to abiotic stress. The increased expression of the Δ12DESATURASE gene and enhanced PLD activity lead to higher level of linoleic acid (18:2), which is subsequently oxidized by LOX, resulting in membrane damage and malondialdehyde (MDA) accumulation. Oxidative stress elevates the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), while the conversion of FFAs into TAGs provides protection against ROS. This research offers valuable molecular and biochemical candidates with significant potential to enhance drought tolerance . It enables innovative strategies in lupine breeding and crop improvement to address critical agricultural challenges.</p>","PeriodicalId":9198,"journal":{"name":"BMC Plant Biology","volume":null,"pages":null},"PeriodicalIF":4.3,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11539565/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142589762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
ZmHsp18 screened from the ZmHsp20 gene family confers thermotolerance in maize. 从 ZmHsp20 基因家族筛选出的 ZmHsp18 赋予玉米耐热性。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-06 DOI: 10.1186/s12870-024-05763-5
Ming Xue, Yiwen You, Luyao Zhang, Jinming Cao, Mingliang Xu, Saihua Chen

Heat stress has become one of the abiotic stresses that pose an increasing threat to maize production due to global warming. The Hsp20 gene family confers tolerance to various abiotic stresses in plants. However, very few Hsp20s have been identified in relation to maize thermotolerance. In this study, we conducted a comprehensive study of Hsp20s involved in thermotolerance in maize. A total of 33 maize Hsp20 genes (ZmHsp20s) were identified through scanning for a conserved α-crystalline domain (ACD), and they were categorized into 14 subfamilies based on phylogenetic analysis. These genes are distributed across all maize chromosomes and nine of them are in regions previously identified as heat-tolerance quantitative trait loci (hrQTL). These hrQTL-associated ZmHsp20s show variation in tissue-specific expression profiles under normal conditions, and seven of them possess 1-5 heat stress elements in their promoters. The integration of RNA-seq data with real-time RT-PCR analysis indicated that ZmHsp23.4, ZmHsp22.8B and ZmHsp18 were dramatically induced under heat stress. Additionally, these genes exhibited co-expression patterns with key ZmHsfs, which are crucial in the heat tolerance pathway. When a null mutant carrying a frame-shifted ZmHsp18 gene was subjected to heat stress, its survival rate decreased significantly, indicating a critical role of ZmHsp18 in maize thermotolerance. Our study lays the groundwork for further research into the roles of ZmHsp20s in enhancing maize's thermotolerance.

由于全球变暖,热胁迫已成为对玉米生产构成日益严重威胁的非生物胁迫之一。Hsp20 基因家族赋予植物对各种非生物胁迫的耐受性。然而,与玉米耐热性相关的 Hsp20 基因却很少被发现。在本研究中,我们对参与玉米耐热性的 Hsp20 进行了全面研究。通过扫描保守的α-结晶结构域(ACD),共鉴定出 33 个玉米 Hsp20 基因(ZmHsp20s),并根据系统发育分析将其分为 14 个亚科。这些基因分布在玉米的所有染色体上,其中九个基因位于先前被鉴定为耐热定量性状位点(hrQTL)的区域。这些与 hrQTL 相关的 ZmHsp20s 在正常条件下表现出组织特异性表达谱的变化,其中 7 个基因的启动子中含有 1-5 个热胁迫元件。RNA-seq数据与实时RT-PCR分析的整合表明,ZmHsp23.4、ZmHsp22.8B和ZmHsp18在热胁迫下被显著诱导。此外,这些基因与耐热途径中至关重要的关键 ZmHsfs 呈现出共表达模式。当携带帧移位 ZmHsp18 基因的无效突变体受到热胁迫时,其存活率显著下降,这表明 ZmHsp18 在玉米耐热性中起着关键作用。我们的研究为进一步研究ZmHsp20s在增强玉米耐热性中的作用奠定了基础。
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引用次数: 0
The genetics of fruit skin separation in date palm. 枣椰树果皮分离的遗传学。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-06 DOI: 10.1186/s12870-024-05713-1
Shameem Younuskunju, Yasmin A Mohamoud, Lisa Sara Mathew, Klaus F X Mayer, Karsten Suhre, Joel A Malek

The physical appearance of date palm (Phoenix dactylifera) fruit (dates) is important for its market value. Many date-producing countries experience significant financial losses due to the poor appearance of the fruit, skin separation or puffiness being a major reason. Previous research showed evidence linking the skin separation phenotype to environmental conditions. To investigate this further, a genome-wide association study was conducted using genome data from 199 samples collected from 14 countries. Here, we identified nine genetic loci associated with this phenotype and investigated genes in these regions that may contribute to the phenotype overall. Multiple genes in the associated regions have functional responses to growth regulators and are involved in cell wall development and modification. Analysis of gene expression data shows many are expressed during fruit development. We show that there are both environmental and genetic contributions to the fruit skin separation phenotype. Our results indicate that different date cultivars exhibit varying degrees of skin separation despite genetic similarities or differences. However, genetically different cultivars show extreme differences compared to the phenotype variation between genetically similar cultivars. We demonstrate that beyond environmental factors, genetics is a strong contributor to the most extreme skin separation in some cultivars. Identifying the genetic factors may help better understand the biology and pathways that lead to the environmental effects on skin separation and improve commercial date production. In conclusion, our key finding is that both environmental and genetic factors contribute to skin separation variation, and improvements in environmental factors alone cannot overcome the extreme level of variation observed in some cultivars.

椰枣(Phoenix dactylifera)果实(椰枣)的外观对其市场价值很重要。许多椰枣生产国因果实外观不佳而遭受重大经济损失,果皮分离或浮肿是主要原因。以前的研究表明,果皮分离表型与环境条件有关。为了进一步研究这个问题,我们利用从 14 个国家收集的 199 个样本的基因组数据进行了全基因组关联研究。在这里,我们确定了与这种表型相关的九个基因位点,并调查了这些区域中可能导致这种表型的基因。相关区域的多个基因对生长调节剂有功能反应,并参与细胞壁的发育和修饰。基因表达数据分析显示,许多基因在果实发育过程中表达。我们的研究表明,果皮分离表型的形成既有环境因素,也有遗传因素。我们的研究结果表明,尽管基因上存在相似或差异,但不同的枣树栽培品种会表现出不同程度的果皮分离现象。然而,与基因相似的栽培品种之间的表型差异相比,基因不同的栽培品种表现出极大的差异。我们证明,除了环境因素外,遗传也是导致某些栽培品种出现最极端表皮分离现象的一个重要因素。确定遗传因素可能有助于更好地理解导致环境影响表皮分离的生物学和途径,并提高商业化枣类生产。总之,我们的主要发现是,环境和遗传因素都会导致表皮分离的变化,而仅仅改善环境因素并不能克服在某些栽培品种中观察到的极端变化水平。
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引用次数: 0
Identifying genetic determinants of forage sorghum [Sorghum bicolor (Moench)] adaptation through GWAS. 通过 GWAS 确定牧草高粱 [Sorghum bicolor (Moench)] 适应性的遗传决定因素。
IF 4.3 2区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-05 DOI: 10.1186/s12870-024-05754-6
Partha Pratim Behera, Avinash Singode, B Venkatesh Bhat, Nayanmoni Borah, Harendra Verma, Patel Supriya, Ramendra Nath Sarma

Background: Forage sorghum is a highly valued crop in livestock feed production due to its versatility, adaptability, high productivity, and resilience under adverse environmental conditions, making it a crucial option for sustainable forage production. This study aimed to investigate ninety-five forage sorghum genotypes and identify the marker - trait associations (MTAs) in adaptive traits, including yield and flowering through genome-wide association studies (GWAS).

Results: Using 41,854 polymorphic SNPs, a GWAS involving the GLM, MLM, and FarmCPU models was performed to analyse fourteen adaptive traits. The population structure revealed the presence of two subpopulation groups. Linkage disequilibrium (LD) plots showed varying degrees of LD decay across the chromosomes, with an average LD decay of 19.49 kbp. Twelve common significant QTNs, encoding 17 putative candidate genes, were simultaneously co-detected and studied by at least two or more GWAS methods. Three QTNs were associated to days to 50% flowering; two each to leaf-to-stem ratio and number of nodes per plant; and one each to plant height, leaf width, number of leaves per plant, stem girth, and internodal length. Six candidate genes were associated with days to 50% flowering, two each with leaf width, stem girth, leaf-to-stem ratio, and number of nodes per plant, and one each with plant height, number of leaves per plant, and internodal length.

Conclusion: FarmCPU was identified as the most suitable and effective among all the models for controlling both false positives and false negatives. Further in-depth analysis of the newly discovered QTNs may lead to the identification of new candidate genes for the trait of interest. These studies elucidate gene functions and could transform forage sorghum breeding through marker-assisted selection and transgenic approaches, accelerating the development of superior forage sorghum varieties and enhancing global food security.

背景:饲用高粱因其用途广泛、适应性强、产量高以及在不利环境条件下的恢复能力强而成为牲畜饲料生产中备受重视的作物,是可持续饲草生产的重要选择。本研究旨在调查 95 个饲用高粱基因型,并通过全基因组关联研究(GWAS)确定产量和开花等适应性性状的标记-性状关联(MTAs):利用 41,854 个多态 SNPs,采用 GLM、MLM 和 FarmCPU 模型进行了全基因组关联研究,分析了 14 个适应性性状。种群结构显示存在两个亚种群组。连锁不平衡(LD)图显示,各染色体上的 LD 有不同程度的衰减,平均 LD 衰减为 19.49 kbp。通过至少两种或两种以上的 GWAS 方法,同时共同检测和研究了 12 个常见的重要 QTNs,它们编码 17 个推测候选基因。3个QTN与50%开花天数有关;2个QTN与叶茎比和每株节数有关;1个QTN与株高、叶宽、每株叶片数、茎围和节间长度有关。6个候选基因与50%开花天数有关,2个候选基因与叶宽、茎围、叶茎比和每株节数有关,1个候选基因与株高、每株叶片数和节间长度有关:结论:在所有模型中,FarmCPU 被认为是最合适、最有效的假阳性和假阴性控制模型。对新发现的 QTN 进一步深入分析,可能会发现相关性状的新候选基因。这些研究阐明了基因功能,可通过标记辅助选择和转基因方法改变饲用高粱育种,加速优良饲用高粱品种的开发,提高全球粮食安全。
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引用次数: 0
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BMC Plant Biology
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