Genome-wide

全基因组
  • 文章类型: Journal Article
    bZIP基因(bZIP)在许多生物过程中是必不可少的,包括发展和应激反应。尽管在许多植物中对BZIP进行了广泛的研究,尚未对大蒜中的bZIP进行全面的全基因组分析。
    在这项研究中,我们鉴定并将64个AsbZIP基因(AsbZIPs)分为10个亚家族。系统分析了这些AsbZIP的进化特征,包括染色体位置,基因结构,保守的图案,和基因复制,进行了。此外,我们还检查了核苷酸多样性,顺式作用元素,和AsbZIPs在各种组织中以及在不同非生物胁迫和激素处理下的表达谱。
    我们的发现表明,基因复制在AsbZIP的扩增中起着至关重要的作用,在驯化过程中观察到轻微的遗传瓶颈。此外,顺式作用元素的鉴定表明AsbZIP与大蒜发育的潜在关联,激素,和应激反应。几个AsbZIP表现出组织偏好和应激/激素响应表达模式。此外,Asa7G01972和Asa7G01379在各种胁迫和激素处理下显著差异表达。随后的酵母两次杂交和酵母诱导实验验证了它们与ABI5的同源物Asa1G01577的相互作用,增强了它们在激素和非生物应激反应中的重要性。本研究揭示了AsbZIP超家族的特征,为进一步分析大蒜中AsbZIP的功能奠定了坚实的基础。
    UNASSIGNED: The bZIP genes (bZIPs) are essential in numerous biological processes, including development and stress responses. Despite extensive research on bZIPs in many plants, a comprehensive genome-wide analysis of bZIPs in garlic has yet to be undertaken.
    UNASSIGNED: In this study, we identified and classified 64 AsbZIP genes (AsbZIPs) into 10 subfamilies. A systematic analysis of the evolutionary characteristics of these AsbZIPs, including chromosome location, gene structure, conserved motifs, and gene duplication, was conducted. Furthermore, we also examined the nucleotide diversity, cis-acting elements, and expression profiles of AsbZIPs in various tissues and under different abiotic stresses and hormone treatments.
    UNASSIGNED: Our findings revealed that gene replication plays a crucial role in the expansion of AsbZIPs, with a minor genetic bottleneck observed during domestication. Moreover, the identification of cis-acting elements suggested potential associations of AsbZIPs with garlic development, hormone, and stress responses. Several AsbZIPs exhibited tissue-preferential and stress/hormone-responsive expression patterns. Additionally, Asa7G01972 and Asa7G01379 were notably differentially expressed under various stresses and hormone treatments. Subsequent yeast two-hybridization and yeast induction experiments validated their interactions with Asa1G01577, a homologue of ABI5, reinforcing their importance in hormone and abiotic stress responses. This study unveiled the characteristics of the AsbZIP superfamily and lays a solid foundation for further functional analysis of AsbZIP in garlic.
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  • 文章类型: Journal Article
    染色质内源性切割与高通量测序(ChEC-seq)是一种蛋白质-DNA相互作用的分析方法,可以在体内检测结合位置,不需要抗体或固定,并提供接近核苷酸分辨率的全基因组覆盖。该方法的核心是目标蛋白的MNase融合,这允许它,当被钙暴露触发时,在其结合位点切割DNA并产生小的DNA片段,这些片段可以很容易地与基因组的其余部分分离并测序。自原始协议以来的改进增加了易用性,降低了成本,并乘以该方法的吞吐量,以实现ChIP-seq等传统方法无法实现的实验规模和分辨率。该方法描述了从MNase标记的酵母菌株的初始创建和验证的每个步骤,通过ChECMNase活化和小片段纯化程序进行测序文库制备。它还简要介绍了创建有意义的全基因组结合谱所需的生物信息学步骤。
    Chromatin endogenous cleavage coupled with high-throughput sequencing (ChEC-seq) is a profiling method for protein-DNA interactions that can detect binding locations in vivo, does not require antibodies or fixation, and provides genome-wide coverage at near nucleotide resolution.The core of this method is an MNase fusion of the target protein, which allows it, when triggered by calcium exposure, to cut DNA at its binding sites and to generate small DNA fragments that can be readily separated from the rest of the genome and sequenced.Improvements since the original protocol have increased the ease, lowered the costs, and multiplied the throughput of this method to enable a scale and resolution of experiments not available with traditional methods such as ChIP-seq. This method describes each step from the initial creation and verification of the MNase-tagged yeast strains, over the ChEC MNase activation and small fragment purification procedure to the sequencing library preparation. It also briefly touches on the bioinformatic steps necessary to create meaningful genome-wide binding profiles.
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  • 文章类型: Journal Article
    在减数分裂期间,spo11产生DNA双链断裂诱导重组,在此过程中,共价连接到断裂两侧的5'末端。这种“共价复合物”在野生型细胞中是瞬时的,但积累的核酸酶突变体无法启动修复。此处提供的CC-seq方法详细介绍了如何在同步酿酒酵母减数分裂细胞中以链特异性核苷酸分辨率精度在全基因组范围内绘制这些Po11复合物的位置。
    During meiosis, Spo11 generates DNA double-strand breaks to induce recombination, becoming covalently attached to the 5\' ends on both sides of the break during this process. Such Spo11 \"covalent complexes\" are transient in wild-type cells, but accumulate in nuclease mutants unable to initiate repair. The CC-seq method presented here details how to map the location of these Spo11 complexes genome-wide with strand-specific nucleotide-resolution accuracy in synchronized Saccharomyces cerevisiae meiotic cells.
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  • 文章类型: Journal Article
    香蕉是热带和亚热带地区最重要的经济作物之一。干旱和低温胁迫影响香蕉的生长。DREB(脱水反应元件结合蛋白)基因家族,作为主要的转录因子家族之一,在防御非生物胁迫中起着至关重要的作用。目前,尚未报道对香蕉DREB(MaDREB)基因家族的系统分析。在这项研究中,在香蕉基因组中鉴定出MaDREB基因家族的103个成员。此外,转录组学分析结果表明,MaDREBs对干旱和寒冷胁迫有反应。干旱和寒冷胁迫诱导了MaDREB14/22/51的表达;选择这些基因进行进一步分析。qRT-PCR验证结果证实了转录组结果。此外,过表达MaDREB14/22/51的转基因拟南芥植物通过降低MDA含量,增加PRO和可溶性糖含量,表现出对干旱和寒冷胁迫的抵抗力。这项研究增强了我们对MaDREB基因家族功能的理解,提供了对它们在非生物胁迫下的调节作用的新见解,为提高香蕉耐旱性和耐寒性奠定了良好的基础。
    Bananas are one of the most important cash crops in the tropics and subtropics. Drought and low-temperature stress affect the growth of banana. The DREB (dehydration responsive element binding protein) gene family, as one of the major transcription factor families, plays crucial roles in defense against abiotic stress. Currently, systematic analyses of the banana DREB (MaDREB) gene family have not yet been reported. In this study, 103 members of the MaDREB gene family were identified in the banana genome. In addition, transcriptomic analysis results revealed that MaDREBs responded to drought and cold stress. The expression of MaDREB14/22/51 was induced by drought and cold stress; these geneswere selected for further analysis. The qRT-PCR validation results confirmed the transcriptome results. Additionally, transgenic Arabidopsis plants overexpressing MaDREB14/22/51 exhibited enhanced resistance to drought and cold stress by reducing MDA content and increasing PRO and soluble sugar content. This study enhances our understanding of the function of the MaDREB gene family, provides new insights into their regulatory role under abiotic stress, and lays a good foundation for improving drought and cold stress-tolerant banana verities.
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  • 文章类型: Journal Article
    腺苷激酶(adenosinekinase,ADK)是一种广泛分布于植物体内的关键酶,在维持细胞能量稳态和调节植物生长方面发挥着重要作用,发展,以及对环境压力的反应。然而,棉花ADK基因的研究,经济意义重大的作物,是有限的。这项研究鉴定了来自四个棉花物种的92个ADK基因(G。Arboreum,G.raimondii,G.hirsutum,和G.barbadense)使用HMMER和LocalBLASTP方法,并将其分为六组。染色体定位揭示了陆地棉中ADK基因的随机分布,有13个基因位于At亚基因组上,14个基因位于Dt亚基因组上。基因结构分析显示亚组内外显子-内含子组织的一致性,虽然保守的基序分析确定了亚组特异性基序,表明功能多样性。合成和共线性作图分析表明,棉花ADK基因家族的主要扩展机制是多倍体和节段复制。GhADK启动子中的顺式调控元件被分类为光响应,激素反应,发育调节,和应激反应。我们还分析了GhADK基因在低温(4°C)和干旱条件下的表达模式。大多数GhADK基因对寒冷胁迫的反应具有不同的表达模式,表明它们在快速反应和长期冷适应中的作用。在干旱胁迫下,表达模式各不相同,一些基因显示出持续的高表达水平。转录组数据的qRT-PCR验证证实了所选GhADK基因的应激诱导表达模式。通过GhADK25的VIGS沉默的功能分析证明了其在寒冷和干旱胁迫反应中的重要性。沉默导致压力下生长不良,强调其在抗逆性中的意义。本研究为进一步了解棉花ADK基因家族的进化关系和功能提供了基础。
    Adenosine kinase (ADK) is a key enzyme widely distributed in plants, playing an important role in maintaining cellular energy homeostasis and regulating plant growth, development, and responses to environmental stresses. However, research on ADK genes in cotton (Gossypium hirsutum), an economically significant crop, has been limited. This study identified 92 ADK genes from four cotton species (G. arboreum, G. raimondii, G. hirsutum, and G. barbadense) using HMMER and Local BLASTP methods and classified them into six groups. Chromosomal localization revealed a random distribution of ADK genes in G. hirsutum, with 13 genes located on the At subgenome and 14 genes on the Dt subgenome. Gene structure analysis showed consistency in exon-intron organization within subgroups, while conserved motif analysis identified subgroup-specific motifs, indicating functional diversity. Synteny and collinearity mapping analysis revealed that the primary expansion mechanisms of the ADK gene family in cotton are polyploidy and segmental duplication. Cis-regulatory elements in GhADK promoters were classified into light response, hormone response, developmental regulation, and stress response. We also analyzed the expression patterns of GhADK genes under a low temperature (4 °C) and drought conditions. Most GhADK genes responded to cold stress with different expression patterns, indicating their roles in rapid response and long-term cold adaptation. Under drought stress, expression patterns varied, with some genes showing sustained high expression levels. The qRT-PCR validation of transcriptomic data confirmed the stress-induced expression patterns of selected GhADK genes. Functional analysis through the VIGS silencing of GhADK25 demonstrated its importance in cold and drought stress responses, with silencing resulting in poor growth under stress, highlighting its significance in stress tolerance. This study provides a basis for further understanding the evolutionary relationships and functions of the cotton ADK gene family.
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  • 文章类型: Journal Article
    栽培花生(ArachishypogaeaL.)是世界上主要的提供石油和蛋白质的豆类作物。它充满了营养,它的营养成分与其他坚果相当。花生是一种独特的植物,因为它表现出钉住现象,地上产花,受精后,发育中的钉子进入土壤并在地下产生种子。花生的这种地皮性质使其种子暴露于土壤病原体。花生种子由不可食用的果皮和种皮保护。果皮和果皮特异性启动子可以有效地用于提高种子防御能力。我们从可用的转录组表达数据中确定了果皮和睾丸丰富的表达基因(AhN8DT-2),其组织特异性表达通过qRT-PCR进一步证实。使用pMDC164载体使用1827bp启动子序列构建表达载体用于进一步分析。GUS基因在转基因拟南芥植株中的定量表达显示其在果皮中的高表达。GUS染色显示果皮和种皮呈深蓝色。染色的拟南芥种子的低温切片显示,表达仅限于种皮(testa),子叶和胚胎中不存在染色。在任何其他组织中均未检测到GUS染色,包括幼苗,叶子,茎,和根,除了花朵上的一些染色。在不同的植物激素下,该启动子没有显示表达水平的增加。这些结果表明AhN8DT-2启动子以果皮和果皮特异性方式驱动GUS基因表达。鉴定的启动子可用于驱动抗病基因,特别是在果皮和果皮中,增强花生种子对土传病原体的防御能力。这种方法对提高花生作物和其他豆类作物的抗逆性具有更广泛的影响,促进可持续农业实践和粮食安全。
    Cultivated peanut (Arachis hypogaea L.) is a key oil- and protein-providing legume crop of the world. It is full of nutrients, and its nutrient profile is comparable to that of other nuts. Peanut is a unique plant as it showcases a pegging phenomenon, producing flowers above ground, and after fertilization, the developing peg enters the soil and produces seeds underground. This geocarpic nature of peanut exposes its seeds to soil pathogens. Peanut seeds are protected by an inedible pericarp and testa. The pericarp- and testa-specific promoters can be effectively used to improve the seed defense. We identified a pericarp- and testa-abundant expression gene (AhN8DT-2) from available transcriptome expression data, whose tissue-specific expression was further confirmed by the qRT-PCR. The 1827bp promoter sequence was used to construct the expression vector using the pMDC164 vector for further analysis. Quantitative expression of the GUS gene in transgenic Arabidopsis plants showed its high expression in the pericarp. GUS staining showed a deep blue color in the pericarp and testa. Cryostat sectioning of stained Arabidopsis seeds showed that expression is only limited to seed coat (testa), and staining was not present in cotyledons and embryos. GUS staining was not detected in any other tissues, including seedlings, leaves, stems, and roots, except for some staining in flowers. Under different phytohormones, this promoter did not show an increase in expression level. These results indicated that the AhN8DT-2 promoter drives GUS gene expression in a pericarp- and testa-specific manner. The identified promoter can be utilized to drive disease resistance genes, specifically in the pericarp and testa, enhancing peanut seed defense against soil-borne pathogens. This approach has broader implications for improving the resilience of peanut crops and other legumes, contributing to sustainable agricultural practices and food security.
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  • 文章类型: Journal Article
    蜘蛛螨侵染对茶叶生长和品质有严重影响。在这项研究中,我们利用染色体定位和共线性分析对CsPIP5K基因家族的功能和调控进行了深入的探索。此外,我们仔细研究了这些基因中的顺式元素。为了充分了解CsPIP5K在蜘蛛螨感染下的代谢反应,我们整合了以前发表的代谢组和转录组数据.我们的分析表明,多个CsPIP5K基因与磷脂代谢有关,与CsPIP5K06显示最强的相关性。因此,我们使用qPCR和亚细胞定位技术来确定该基因的表达模式及其在细胞中的功能位置。总的来说,这项研究不仅全面阐明了特点,结构,和CsPIP5K基因家族的进化,但也根据先前发表的数据确定了与磷脂生物合成相关并与蜘蛛螨相关的几个候选CsPIP5K基因。这项研究为提高茶叶对蜘蛛螨的抗性和保持最佳茶叶品质做出了重大贡献。
    Spider mite infestation has a severe impact on tea growth and quality. In this study, we conducted a deep exploration of the functions and regulations of the CsPIP5K gene family using chromosomal localization and collinearity analysis. Additionally, we carefully examined the cis elements within these genes. To fully understand the metabolic response of CsPIP5K under spider mite infection, we integrated previously published metabolomic and transcriptomic data. Our analysis revealed that multiple CsPIP5K genes are associated with phospholipid metabolism, with CsPIP5K06 showing the strongest correlation. Therefore, we employed qPCR and subcellular localization techniques to determine the expression pattern of this gene and its functional location within the cell. Overall, this study not only comprehensively elucidated the characteristics, structure, and evolution of the CsPIP5K gene family but also identified several candidate CsPIP5K genes related to phospholipid biosynthesis and associated with spider mites based on previously published data. This research makes a significant contribution to enhancing the resistance of tea to spider mite and maintaining optimal tea quality.
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  • 文章类型: Journal Article
    本研究的重点是分析大麻UGT基因家族,在植物的代谢和次生代谢产物的糖基化中起着至关重要的作用。该研究使用保守的植物次级产物糖基转移酶(PSPG)基序氨基酸序列鉴定了125个UGT。这些UGT基因通过系统发育分析分为17组(A-Q),显示它们在水稻10条染色体上的分布。CsUGT基因家族的扩展归因于串联和重复事件,正如基因复制分析所表明的。此外,研究发现CsUGT启动子区存在多种与植物激素和应激反应相关的顺式调控元件。亚细胞定位分析显示CsUGT存在于细胞质中,叶绿体,和核。研究表明,CsUGT在各种生物过程中发挥着重要作用,细胞成分,和基因本体论分析强调的分子功能。此外,京都基因和基因组百科全书(KEGG)途径分析表明,一些CsUGTs与次级代谢产物的生物合成有关。这项研究为基因组组织提供了有价值的见解,进化史,以及紫花苜蓿UGT基因的潜在调控机制。为进一步探索它们在植物代谢和逆境响应中的具体生物学作用和潜在应用奠定了基础。这些发现有助于更好地理解UGT基因家族及其与水稻代谢途径的相关性。
    在线版本包含补充材料,可在10.1007/s13205-024-04025-3获得。
    The research focused on analyzing the UGT gene family in Cannabis sativa, which plays a crucial role in the plant\'s metabolism and glycosylation of secondary metabolites. The study identified 125 UGTs using conserved plant secondary product glycosyltransferase (PSPG) motif amino acid sequences. These UGT genes were categorized into 17 groups (A-Q) through phylogenetic analysis, showing their distribution across 10 chromosomes in C. sativa. The expansion of the CsUGT gene family was attributed to tandem and duplication events, as suggested by gene duplication analysis. Furthermore, the study found various cis-acting regulatory elements related to phytohormones and stress responses in CsUGT promoter regions. Subcellular localization analysis revealed that CsUGT is present in the cytoplasm, chloroplast, and nucleus. The study revealed that CsUGT plays a significant role in various biological processes, cellular components, and molecular functions as highlighted by Gene Ontology analysis. Additionally, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that some CsUGTs are associated with the biosynthesis of secondary metabolites. This research provides valuable insights into the genomic organization, evolutionary history, and potential regulatory mechanisms of UGT genes in C. sativa. It lays the foundation for further exploration of their specific biological roles and potential applications in the plant\'s metabolism and stress responses. These findings contribute to a better understanding of the UGT gene family and its relevance to the metabolic pathways in C. sativa.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s13205-024-04025-3.
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  • 文章类型: Journal Article
    非生物胁迫是花生生产中的限制因素。花生是我国重要的油料作物和经济作物。花生的产量由于其种子生长在地下而容易受到非生物胁迫。茉莉酸(JA)对于植物生长和抵抗逆境胁迫至关重要。然而,茉莉酸生物合成途径对花生防御非生物胁迫的调控和机制仍然有限。在这项研究中,共有64个编码JA生物合成关键酶的基因被鉴定并分类为脂氧合酶(AhLOXs),氧化烯合酶(AhAOSs),丙二烯氧化物环化酶(AhAOCs),和根据基因结构的12-氧代-植物二烯酸还原酶(AhOPR),保守的图案,和系统发育特征。顺式调节元件分析表明,某些基因包含应激反应和激素反应元件。除了参与JA生物合成和信号传导的蛋白质,它们还与参与脂质生物合成和应激反应的蛋白质相互作用。从靶向JA生物合成的35个关键基因的4个家族中鉴定出16个推定的Ah-miRNA。组织表达模式分析显示AhLOX2在叶片组织中表达量最高,AhLOX32在枝条中表达最高,根,和结节组织。AhLOX16、AhOPR1和AhOPR3在干旱胁迫下上调。AhLOX16、AhAOS3、AhOPR1和AhAOC4响应冷应激具有升高的转录水平。AhLOX5、AhLOX16、AhAOC3、AhOPR1和AhOPR3在盐胁迫下表达上调。本研究可为花生非生物抗逆机制的研究提供参考。
    Abiotic stress is a limiting factor in peanut production. Peanut is an important oil crop and cash crop in China. Peanut yield is vulnerable to abiotic stress due to its seeds grown underground. Jasmonic acid (JA) is essential for plant growth and defense against adversity stresses. However, the regulation and mechanism of the jasmonic acid biosynthesis pathway on peanut defense against abiotic stresses are still limitedly understood. In this study, a total of 64 genes encoding key enzymes of JA biosynthesis were identified and classified into lipoxygenases (AhLOXs), alleno oxide synthases (AhAOSs), allene oxide cyclases (AhAOCs), and 12-oxo-phytodienoic acid reductases (AhOPRs) according to gene structure, conserved motif, and phylogenetic feature. A cis-regulatory element analysis indicated that some of the genes contained stress responsive and hormone responsive elements. In addition to proteins involved in JA biosynthesis and signaling, they also interacted with proteins involved in lipid biosynthesis and stress response. Sixteen putative Ah-miRNAs were identified from four families targeting 35 key genes of JA biosynthesis. A tissue expression pattern analysis revealed that AhLOX2 was the highest expressed in leaf tissues, and AhLOX32 was the highest expressed in shoot, root, and nodule tissues. AhLOX16, AhOPR1, and AhOPR3 were up-regulated under drought stress. AhLOX16, AhAOS3, AhOPR1, and AhAOC4 had elevated transcript levels in response to cold stress. AhLOX5, AhLOX16, AhAOC3, AhOPR1, and AhOPR3 were up-regulated for expression under salt stress. Our study could provide a reference for the study of the abiotic stress resistance mechanism in peanut.
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  • 文章类型: Journal Article
    糖最终将外溢的转运蛋白(SWEET)已被证实在植物生长中起着不同的生理作用,发育和应激反应。然而,Hemerocalliscitrina中SWEET基因的特征和功能仍不清楚,也不清楚。在这项研究中,利用全基因组进行生物信息学分析,成功鉴定出19个HcSWEET基因。物理化学性质的分析表明这些HcSWEET之间存在主要差异。系统发育分析表明,HcSWEET蛋白可以分为4个进化枝,从进化枝I到IV,其中同一进化枝内的蛋白质表现出共享的保守基序和基因结构。大多数HcSWEET基因中含有5到6个外显子,它们在11条染色体上分布不均。基因重复分析显示存在4个基因对。比较性图谱显示,与双子叶植物相比,HcSWEET基因家族在单子叶植物中可能表现出更多的封闭同源性。HcSWEET基因的顺式作用元件分析表明对各种激素的关键反应,光,和压力。此外,转录组测序分析表明,大多数HcSWEET基因在根中的表达相对较高,HcSWEET4a在盐胁迫下显著上调。过表达进一步证实了HcSWEET4a参与盐胁迫反应的可能性,这提供了新颖的见解,并促进了对HcSWEET在抵抗非生物胁迫方面的功能分析的深入研究。
    Sugars will be eventually effluxed transporters (SWEETs) have been confirmed to play diverse physiological roles in plant growth, development and stress response. However, the characteristics and functions of the SWEET genes in Hemerocallis citrina remain unclear and poorly elucidated. In this study, the whole genome of Hemerocallis citrina was utilized to conduct bioinformatics analysis and a total of 19 HcSWEET genes were successfully identified. Analysis of the physicochemical properties indicated dominant differences among these HcSWEETs. A phylogenetic analysis revealed that HcSWEET proteins can be divided into 4 clades ranging from Clade I to IV, where proteins within the same clade exhibited shared conserved motifs and gene structures. Five to six exons were contained in the majority of HcSWEET genes, which were unevenly distributed across 11 chromosomes. The gene duplication analysis showed the presence of 4 gene pairs. Comparative syntenic maps revealed that the HcSWEET gene family might present more closed homology in monocotyledons than dicotyledons. Cis-acting element analysis of HcSWEET genes indicated key responsiveness to various hormones, light, and stresses. Additionally, transcriptome sequencing analysis suggested that most HcSWEET genes had a relatively higher expression in roots, and HcSWEET4a was significantly up-regulated under salt stress. Overexpression further verified the possibility that HcSWEET4a was involved in response to salt stress, which provides novel insights and facilitates in-depth studies of the functional analysis of HcSWEETs in resistance to abiotic stress.
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