Plant Direct最新文献

{"title":"Wheat NAC transcription factor <i>NAC5-1</i> is a positive regulator of senescence.","authors":"Catherine Evans, Sophie Louise Mogg, Charline Soraru, Emma Wallington, Juliet Coates, Philippa Borrill","doi":"10.1002/pld3.620","DOIUrl":"10.1002/pld3.620","url":null,"abstract":"<p><p>Wheat (<i>Triticum aestivum</i> L.) is an important source of both calories and protein in global diets, but there is a trade-off between grain yield and protein content. The timing of leaf senescence could mediate this trade-off as it is associated with both declines in photosynthesis and nitrogen remobilization from leaves to grain. NAC transcription factors play key roles in regulating senescence timing. In rice, <i>OsNAC5</i> expression is correlated with increased protein content and upregulated in senescing leaves, but the role of the wheat ortholog in senescence had not been characterized. We verified that <i>NAC5-1</i> is the ortholog of <i>OsNAC5</i> and that it is expressed in senescing flag leaves in wheat. To characterize <i>NAC5-1</i>, we combined missense mutations in <i>NAC5-A1</i> and <i>NAC5-B1</i> from a TILLING mutant population and overexpressed <i>NAC5-A1</i> in wheat. Mutation in <i>NAC5-1</i> was associated with delayed onset of flag leaf senescence, while overexpression of <i>NAC5-A1</i> was associated with slightly earlier onset of leaf senescence. DAP-seq was performed to locate transcription factor binding sites of <i>NAC5-1</i>. Analysis of DAP-seq and comparison with other studies identified putative downstream target genes of <i>NAC5-1</i> which could be associated with senescence. This work showed that <i>NAC5-1</i> is a positive transcriptional regulator of leaf senescence in wheat. Further research is needed to test the effect of <i>NAC5-1</i> on yield and protein content in field trials, to assess the potential to exploit this senescence regulator to develop high-yielding wheat while maintaining grain protein content.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11217990/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141498838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Albino lethal 13, a chloroplast-imported protein required for chloroplast development in rice.","authors":"Xiaoqiong Guo, Chunli Wang, Qian Zhu, Wenhua Dongchen, Xiaoling Zhang, Wei Li, Hui Zhang, Cui Zhang, Zar Ni Naing Nant Nyein, Mengting Li, Lijuan Chen, Dongsun Lee","doi":"10.1002/pld3.610","DOIUrl":"10.1002/pld3.610","url":null,"abstract":"<p><p>Chloroplasts play a vital role in plant growth and development, which are the main sites of photosynthesis and the production of hormones and metabolites. Despite their significance, the regulatory mechanisms governing chloroplast development remain unclear. In our investigation, we identified a rice mutant with defective chloroplasts in rice (<i>Oryza sativa</i> L.), named albino lethal 13 (<i>osal13</i>), which displayed a distinct albino phenotype in leaves, ultimately resulting in seedling lethality. Molecular cloning revealed that <i>OsAL13</i> encodes a novel rice protein with no homologous gene or known conserved domain. This gene was located in the chloroplast and exhibited constitutive expression in various tissues, particularly in green tissues and regions of active cell growth. Our study's findings reveal that RNAi-mediated knockdown of <i>OsAL13</i> led to a pronounced albino phenotype, reduced chlorophyll and carotenoid contents, a vesicle chloroplast structure, and a decrease in the expression of chloroplast-associated genes. Consequently, the pollen fertility and seed setting rate were lower compared with the wild type. In contrast, the overexpression of <i>OsAL13</i> resulted in an increased photosynthetic rate, a higher total grain number per panicle, and enhanced levels of indole-3-acetic acid (IAA) in the roots and gibberellin A3 (GA3) in the shoot. These outcomes provide new insights on the role of <i>OsAL13</i> in regulating chloroplast development in rice.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11189691/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141432624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The activation of <i>Chlamydomonas reinhardtii</i> alpha amylase 2 by glutamine requires its N-terminal aspartate kinase-chorismate mutase-tyrA (ACT) domain.","authors":"Lisa Scholtysek, Ansgar Poetsch, Eckhard Hofmann, Anja Hemschemeier","doi":"10.1002/pld3.609","DOIUrl":"10.1002/pld3.609","url":null,"abstract":"<p><p>The coordination of assimilation pathways for all the elements that make up cellular components is a vital task for every organism. Integrating the assimilation and use of carbon (C) and nitrogen (N) is of particular importance because of the high cellular abundance of these elements. Starch is one of the most important storage polymers of photosynthetic organisms, and a complex regulatory network ensures that biosynthesis and degradation of starch are coordinated with photosynthetic activity and growth. Here, we analyzed three starch metabolism enzymes of <i>Chlamydomonas reinhardtii</i> that we captured by a cyclic guanosine monophosphate (cGMP) affinity chromatography approach, namely, soluble starch synthase STA3, starch-branching enzyme SBE1, and α-amylase AMA2. While none of the recombinant enzymes was directly affected by the presence of cGMP or other nucleotides, suggesting an indirect binding to cGMP, AMA2 activity was stimulated in the presence of L-glutamine (Gln). This activating effect required the enzyme's N-terminal aspartate kinase-chorismate mutase-tyrA domain. Gln is the first N assimilation product and not only a central compound for the biosynthesis of N-containing molecules but also a recognized signaling molecule for the N status. Our observation suggests that AMA2 might be a means to coordinate N and C metabolism at the enzymatic level, increasing the liberation of C skeletons from starch when high Gln levels signal an abundance of assimilated N.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11190351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141443265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Xanthohumol and echinocystic acid induces PSTVd tolerance in tomato.","authors":"Wenkun Tang, Zhichao Tang, Haiyi Liu, Jinbiao Lu, Qianyun Du, Huan Tian, Jingwei Li","doi":"10.1002/pld3.612","DOIUrl":"10.1002/pld3.612","url":null,"abstract":"<p><p>Tomato is a popular vegetable worldwide; its production is highly threatened by infection with the potato spindle tuber viroid (PSTVd). We obtained the full-length genome sequence of previously conserved PSTVd and inoculated it on four genotypes of semi-cultivated tomatoes selected from a local tomato germplasm resource. SC-5, which is a PSTVd-resistant genotype, and SC-96, which is a PSTVd-sensitive genotype, were identified by detecting the fruit yield, plant growth, biomass accumulation, physiological indices, and PSTVd genome titer after PSTVd inoculation. A non-target metabolomics study was conducted on PSTVd-infected and control SC-5 to identify potential anti-PSTVd metabolites. The platform of liquid chromatography-mass spectrometry detected 158 or 123 differential regulated metabolites in modes of positive ion or negative ion. Principal component analysis revealed a clear separation of the global metabolite profile between PSTVd-infected leaves and control regardless of the detection mode. The potential anti-PSTVd compounds, xanthohumol, oxalicine B, indole-3-carbinol, and rosmarinic acid were significantly upregulated in positive ion mode, whereas echinocystic acid, chlorogenic acid, and 5-acetylsalicylic acid were upregulated in negative ion mode. Xanthohumol and echinocystic acid were detected as the most upregulated metabolites and were exogenously applied on PSTVd-diseased SC-96 seedlings. Both xanthohumol and echinocystic acid had instant and long-term inhibition effect on PSTVd titer. The highest reduction of disease symptom was induced by 2.6 mg/L of xanthohumol and 2.0 mg/L of echinocystic acid after 10 days of leaf spraying, respectively. A superior effect was seen on echinocystic acid than on xanthohumol. Our study provides a statistical basis for breeding anti-viroid tomato genotypes and creating plant-originating chemical preparations to prevent viroid disease.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11190350/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141443266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA barcoding of terrestrial invasive plant species in Southwest Michigan.","authors":"Sneha Nath, Joshua T VanSlambrouck, Janelle W Yao, Ashika Gullapalli, Fayyaz Razi, Yan Lu","doi":"10.1002/pld3.615","DOIUrl":"10.1002/pld3.615","url":null,"abstract":"<p><p>Because of the detrimental effects of terrestrial invasive plant species (TIPS) on native species, ecosystems, public health, and the economy, many countries have been actively looking for strategies to prevent the introduction and minimize the spread of TIPS. Fast and accurate detection of TIPS is essential to achieving these goals. Conventionally, invasive species monitoring has relied on morphological attributes. Recently, DNA-based species identification (i.e., DNA barcoding) has become more attractive. To investigate whether DNA barcoding can aid in the detection and management of TIPS, we visited multiple nature areas in Southwest Michigan and collected a small piece of leaf tissue from 91 representative terrestrial plant species, most of which are invasive. We extracted DNA from the leaf samples, amplified four genomic loci (ITS, <i>rbcL</i>, <i>matK</i>, and <i>trnH-psbA</i>) with PCR, and then purified and sequenced the PCR products. After careful examination of the sequencing data, we were able to identify reliable DNA barcode regions for most species and had an average PCR-and-sequencing success rate of 87.9%. We found that the species discrimination rate of a DNA barcode region is inversely related to the ease of PCR amplification and sequencing. Compared with <i>rbcL</i> and <i>matK</i>, ITS and <i>trnH-psbA</i> have better species discrimination rates (80.6% and 63.2%, respectively). When ITS and <i>trnH-psbA</i> are simultaneously used, the species discrimination rate increases to 97.1%. The high species/genus/family discrimination rates of DNA barcoding indicate that DNA barcoding can be successfully employed in TIPS identification. Further increases in the number of DNA barcode regions show little or no additional increases in the species discrimination rate, suggesting that dual-barcode approaches (e.g., ITS + <i>trnH-psbA</i>) might be the efficient and cost-effective method in DNA-based TIPS identification. Close inspection of nucleotide sequences at the four DNA barcode regions among related species demonstrates that DNA barcoding is especially useful in identifying TIPS that are morphologically similar to other species.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11185875/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141420443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome wide association studies on seven yield-related traits of 183 rice varieties in Bangladesh.","authors":"Nilanjan Roy, Acramul Haque Kabir, Nourin Zahan, Shahba Tasmiya Mouna, Sakshar Chakravarty, Atif Hasan Rahman, Md Shamsuzzoha Bayzid","doi":"10.1002/pld3.593","DOIUrl":"10.1002/pld3.593","url":null,"abstract":"<p><p>Rice genetic diversity is regulated by multiple genes and is largely dependent on various environmental factors. Uncovering the genetic variations associated with the diversity in rice populations is the key to breed stable and high yielding rice varieties. We performed genome wide association studies (GWASs) on seven rice yielding traits (grain length, grain width, grain weight, panicle length, leaf length, leaf width, and leaf angle) based on a population of 183 rice landraces of Bangladesh. Our GWASs reveal various chromosomal regions and candidate genes that are associated with different traits in Bangladeshi rice varieties. Noteworthy was the recurrent implication of chromosome 10 in all three grain-shape-related traits (grain length, grain width, and grain weight), indicating its pivotal role in shaping rice grain morphology. Our study also underscores the involvement of transposon gene families across these three traits. For leaf related traits, chromosome 10 was found to harbor regions that are significantly associated with leaf length and leaf width. The results of these association studies support previous findings as well as provide additional insights into the genetic diversity of rice. This is the first known GWAS study on various yield-related traits in the varieties of <i>Oryza sativa</i> available in Bangladesh-the fourth largest rice-producing country. We believe this study will accelerate rice genetics research and breeding stable high-yielding rice in Bangladesh.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11182691/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141420444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of the COPII vesicle-forming GTPase Sar1 in <i>Chlamydomonas</i>.","authors":"Kin Pan Chung, Daniel Frieboese, Florent Waltz, Benjamin D Engel, Ralph Bock","doi":"10.1002/pld3.614","DOIUrl":"10.1002/pld3.614","url":null,"abstract":"<p><p>Eukaryotic cells are highly compartmentalized, requiring elaborate transport mechanisms to facilitate the movement of proteins between membrane-bound compartments. Most proteins synthesized in the endoplasmic reticulum (ER) are transported to the Golgi apparatus through COPII-mediated vesicular trafficking. Sar1, a small GTPase that facilitates the formation of COPII vesicles, plays a critical role in the early steps of this protein secretory pathway. Sar1 was characterized in yeast, animals and plants, but no Sar1 homolog has been identified and functionally analyzed in algae. Here we identified a putative Sar1 homolog (CrSar1) in the model green alga <i>Chlamydomonas reinhardtii</i> through amino acid sequence similarity. We employed site-directed mutagenesis to generate a dominant-negative mutant of CrSar1 (CrSar1DN). Using protein secretion assays, we demonstrate the inhibitory effect of CrSar1DN on protein secretion. However, different from previously studied organisms, ectopic expression of CrSar1DN did not result in collapse of the ER-Golgi interface in <i>Chlamydomonas</i>. Nonetheless, our data suggest a largely conserved role of CrSar1 in the ER-to-Golgi protein secretory pathway in green algae.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11180857/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141420445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A chemical genetic screen with the EXO70 inhibitor Endosidin2 uncovers potential modulators of exocytosis in Arabidopsis.","authors":"Xiaohui Li, Diwen Wang, Xianglin Yin, Mingji Dai, Christopher J Staiger, Chunhua Zhang","doi":"10.1002/pld3.592","DOIUrl":"10.1002/pld3.592","url":null,"abstract":"<p><p>Exocytosis plays an essential role in delivering proteins, lipids, and cell wall polysaccharides to the plasma membrane and extracellular spaces. Accurate secretion through exocytosis is key to normal plant development as well as responses to biotic and abiotic stresses. During exocytosis, an octameric protein complex named the exocyst facilitates the tethering of secretory vesicles to the plasma membrane. Despite some understanding of molecular and cellular aspects of exocyst function obtained through reverse genetics and direct interaction assays, knowledge about upstream modulators and genetic interactors remains limited. Traditional genetic screens encounter practical issues in exocyst subunit mutant backgrounds, such as lethality of certain knockout mutants and/or potential redundancy of EXO70 homologs. To address these challenges, this study leverages the tunable and reversible nature of chemical genetics, employing Endosidin2 (ES2)-a synthetic inhibitor of EXO70-for a large-scale chemical genetic mutant screen in Arabidopsis. This approach led to the identification of 70 ES2-hypersensitive mutants, named <i>es2s</i>. Through a whole-genome sequencing-based mapping strategy, 14 nonallelic <i>es2s</i> mutants were mapped and the candidate mutations reported here. In addition, T-DNA insertion lines were tested as alternative alleles to identify causal mutations. We found that T-DNA insertion alleles for <i>DCP5</i>, <i>VAS1/ISS1</i>, <i>ArgJ</i>, and <i>MEF11</i> were hypersensitive to ES2 for root growth inhibition. This research not only offers new genetic resources for systematically identifying molecular players interacting with the exocyst in Arabidopsis but also enhances understanding of the regulation of exocytosis.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11176578/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141331609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intraspecific differences in the photosynthetic responses to chloroplast CO₂ and photon flux density at different leaf temperatures of four grapevine cultivars grown in common outdoor conditions.","authors":"Dennis H Greer","doi":"10.1002/pld3.595","DOIUrl":"10.1002/pld3.595","url":null,"abstract":"<p><p>Comparative measurements of four <i>Vitis vinifera</i> cultivars were undertaken to assess assimilation tolerance to the high growth temperatures currently pervading Australian and other wine growing regions. The cultivars, cvs. Chardonnay, Merlot, Semillon, and Shiraz, were all grown in common growth conditions, and an hypothesis promulgated genotypic variation in assimilation and in the leaf temperature dependency. Assimilation responses to varying light intensity and to varying chloroplast CO₂ at a range of leaf temperatures (15-45°C) were measured in leaves of each cultivar in mid-summer. Light response curves revealed marked genotype differences in maximum assimilation, but temperature effects also varied. Semillon leaves were most sensitive to temperature, with marked and steep differences in assimilation at different temperatures while Chardonnay and Merlot were least sensitive, with relatively flat responses. There were also marked cultivar differences in response to CO₂ and significant effects of leaf temperature. CO₂-saturated assimilation varied markedly, with Semillon and Merlot leaves most responsive to temperature, although there were differences in optimum temperatures and maximum rates. Chardonnay leaves remained least tolerant, with lowest rates of assimilation across most temperatures. Assimilation at 45°C also separated the cultivars and two cultivars had higher rates than at 15°C while Chardonnay and Merlot leaves had higher rates at 15°C. There were no cultivar differences in the temperature dependency of Ribulose 1,5-bisphosphate (RuBP) carboxylation, but Semillon had a much steeper temperature dependency on RuBP regeneration than the other cultivars. All these responses confirmed the hypothesis and concluded the high-temperature tolerance of Semillon and Shiraz and the poor adaptability of Chardonnay and possibly Merlot to perform in the current high-temperature growth conditions.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11154808/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141296621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Critical parameters for robust <i>Agrobacterium</i>-mediated transient transformation and quantitative promoter assays in <i>Catharanthus roseus</i> seedlings.","authors":"Lauren F Cole-Osborn, Emma Meehan, Carolyn W T Lee-Parsons","doi":"10.1002/pld3.596","DOIUrl":"10.1002/pld3.596","url":null,"abstract":"<p><p><i>Agrobacterium</i>-mediated transient expression methods are widely used to study gene function in both model and non-model plants. Using a dual-luciferase assay, we quantified the effect of <i>Agrobacterium</i>-infiltration parameters on the transient transformation efficiency of <i>Catharanthus roseus</i> seedlings. We showed that transformation efficiency is highly sensitive to seedling developmental state and a pre- and post-infiltration dark incubation and is less sensitive to the <i>Agrobacterium</i> growth stage. For example, 5 versus 6 days of germination in the dark increased seedling transformation efficiency by seven- to eight-fold while a dark incubation pre- and post-infiltration increased transformation efficiency by five- to 13-fold. <i>Agrobacterium</i> in exponential compared with stationary phase increased transformation efficiency by two-fold. Finally, we quantified the variation in our <i>Agrobacterium</i>-infiltration method in replicate infiltrations and experiments. Within a given experiment, significant differences of up to 2.6-fold in raw firefly luciferase (<i>FLUC</i>) and raw <i>Renilla</i> luciferase (<i>RLUC</i>) luminescence occurred in replicate infiltrations. These differences were significantly reduced when FLUC was normalized to RLUC values, highlighting the utility of including a reference reporter to minimize false positives. Including a second experimental replicate further reduced the potential for false positives. This optimization and quantitative validation of <i>Agrobacterium</i> infiltration in <i>C. roseus</i> seedlings will facilitate the study of this important medicinal plant and will expand the application of <i>Agrobacterium</i>-mediated transformation methods in other plant species.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11154794/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141296620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}