Transgenic Research最新文献

{"title":"The high-affinity K⁺ transporter IbHAK5 enhances potassium ion absorption and improves root morphology in sweetpotato (Ipomoea batatas).","authors":"Rong Jin, Peng Zhao, Mengxiao Yan, Ming Liu, Weijuan Fan, Qiangqiang Zhang, Xiaoya Zhu, Jing Wang, Yongchao Yu, Jun Yang, Hongxia Wang, Zhonghou Tang","doi":"10.1007/s11248-025-00437-w","DOIUrl":"https://doi.org/10.1007/s11248-025-00437-w","url":null,"abstract":"<p><p>Potassium is a vital element in sweetpotato that plays important roles during its growth and development. In this study, potassium transporter IbHAK5, which is homologous to Arabidopsis HAK5, was cloned and overexpressed in sweetpotato. IbHAK5 encoded a protein of 739 amino acids and localized in the plasma membrane. Two IbHAK5-overexpressing transgenic lines with the highest expression level of IbHAK5 were screened for K⁺-deficiency stress tolerant assay. Compared with wild type sweetpotato plants, transgenic plants grew well with higher chlorophyll content, and maintain great higher K⁺ contents via decreasing more K⁺ effluxes under low potassium ion (- K⁺) stress condition. Additionally, IbHAK5 can help plants improve root morphology and increase endogenous hormone IAA content under both normal condition and - K⁺ stress, which may result in the increased root K⁺ absorption ability. The results indicated that IbHAK5 play an important role in sweetpotato response to - K⁺ stress, as well as support molecular-assisted breeding with the IbHAK5 gene.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"25"},"PeriodicalIF":2.7,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144120935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phenotypic characteristics of interspecific hybrids between wild and cultivated soybean with and without insect-protected biotechnology traits.","authors":"Duška Stojšin, Bill Duncan, Chen Meng","doi":"10.1007/s11248-025-00443-y","DOIUrl":"https://doi.org/10.1007/s11248-025-00443-y","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Wild soybean (Glycine soja Sieb. and Zucc.) and cultivated soybean (Glycine max (L) Merr.) can cross-pollinate, albeit at a very low frequency, potentially resulting in an interspecific hybrid with a biotechnology trait inherited from the cultivated soybean parent. As part of environmental risk assessment, it is informative to understand the competitiveness potential of these hybrids in natural habitats. The objective of this research was to evaluate the phenotypic characteristics of the interspecific hybrids and compare them with cultivated and wild soybean. Secondly, the comparisons were conducted between the interspecific hybrids with and without an insect-protected (IP) biotechnology trait. Two wild soybean populations were crossed with cultivated soybean containing either MON 87701 or MON 87751, the IP traits developed to control specific lepidopteran pests. Hybrid plants with and without the IP trait and parental entries were evaluated in growth chamber trials for the plant, pollen and seed characteristics. Compared to the parents, the hybrids had intermediate values for most of the measured characteristics. Compared to the wild soybean parent, hybrids had less twining, shorter plants, fewer seeds and pods, thicker stems, bigger seeds and pollen grains, as well as reduced seed dormancy-all due to domestication genes inherited from cultivated soybean. However, when compared to parents, hybrids had significantly reduced pollen viability (51.8-73.3% vs. &gt; 95%) and number of seeds per pod (1.6-1.7 vs. 2.3) indicating partial reproductive sterility due to chromosome interchange between nonhomologous chromosomes. Seed coat color of the F&lt;sub&gt;3&lt;/sub&gt; hybrids segregated in a manner that seems to involve two major genes (I and T) with some level of inter-locus interaction and/or partial dominance resulting in an observed ratio of 9 green (i&lt;sup&gt;i&lt;/sup&gt;-T-): 3 black (iiT-): 3 brown (-itt): 1 yellow (i&lt;sup&gt;i&lt;/sup&gt;i&lt;sup&gt;i&lt;/sup&gt;tt) seeds. A combination of recessive genes (ii from wild and tt from cultivated soybean) resulted in hybrid seeds with defective, cracked seed coat contributing to reduction in seed dormancy. Generally, hybrids with and without the IP traits had comparable performance indicating that the biotechnology IP trait did not have unintended effects on phenotypic characteristics. However, there were some characteristics that differed. The F&lt;sub&gt;3&lt;/sub&gt; hybrids with MON 87751 had significantly lower seed dormancy (8.1%) compared to hybrids without the transgene (15.1%) likely due to the proximity of the transgene to soybean native domestication-related gene and their co-segregation in hybrid generations. In summary, the results obtained in this research suggest that the fitness, overwintering and general ability of hybrids to compete and persist in nature is reduced when compared to wild soybean. The main contributors to lower survival of hybrids are: (i) domestication genes inherited from the soybean parent, (ii) partial sterility","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"24"},"PeriodicalIF":2.7,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12078375/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144080579","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":"Knockout of the Agrobacterium ILVC gene generates a valine-isoleucine auxotrophic strain for plant transformation.","authors":"Yanhui Yang, Jun Lu, Tianrui Xue, Zhenli Cao, Mingjie Li, Zhongyi Zhang, Rong Wang, Tongyu Wu","doi":"10.1007/s11248-025-00442-z","DOIUrl":"https://doi.org/10.1007/s11248-025-00442-z","url":null,"abstract":"<p><p>The Agrobacterium tumefaciens mediated transformation is the prevailing methodology for plant genetic manipulation; however, A. tumefaciens overgrowth is a common constraint in the process. Exploring auxotrophic A. tumefaciens could reduce overgrowth and enhance plant transformation efficiency. The ILVC gene, which encodes the ketol-acid isomeroreductase, is critical for Valine (Val) and isoleucine (Ile) biosynthesis in some microorganisms. However, its function in A. tumefaciens is unclear. To ascertain the function of ILVC and generate an auxotrophic A. tumefaciens, this study employed an allelic exchange to disrupt the ILVC in A. tumefaciens strain GV3101. This resulted in the loss of ketol-acid isomeroreductase activity and the prevention of Val and Ile biosynthesis, creating a dual-auxotrophic GV3101_∆ILVC. Transient expression assays in Nicotiana benthamiana transformation demonstrated that the GV3101_∆ILVC was capable of T-DNA transfer. Moreover, stable genetic transformation analysis in N. benthamiana indicated that the introduction of GV3101_∆ILVC led to a reduction in overgrowth within infected plant tissues. Additionally, an enhancement in transformation efficiency was observed with the prolongation of the co-cultivation time of the explant-infected strain. This study revealed the function of ILVC and explored a dual-auxotrophic A. tumefaciens for Val and Ile, potentially broadening the utilization of auxotrophic strains in plant genetic transformation.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"23"},"PeriodicalIF":2.7,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144020042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a plant-based vaccine against brucellosis: stable expression of Brucella abortus OMP25 in transgenic tobacco.","authors":"Mansoure Qashqai, Emrah Bertan, Semiha Erisen, Tulin Ozbek, Senay Vural-Korkut","doi":"10.1007/s11248-025-00441-0","DOIUrl":"https://doi.org/10.1007/s11248-025-00441-0","url":null,"abstract":"<p><p>Brucellosis, caused by Brucella species, is a global threat to livestock farming, resulting in economic losses and socio-economic challenges, particularly in rural areas. Despite its impact, no licensed human vaccines are available. Animal vaccination remains the most cost-effective control method, but traditional vaccine production is expensive. Edible vaccines, using plants as bioreactors to produce immunogenic antigens, offer a low-cost alternative by eliminating complex purification processes. This study developed a transgenic plant by expressing the Brucella abortus outer membrane protein OMP25 in tobacco plants. OMP25, a conserved transmembrane protein with high immunogenicity, was cloned into a Gateway pDONR vector via a Boundary Pairing reaction and transferred to a binary destination vector via a Left-Right reaction. The destination vector was introduced into Agrobacterium tumefaciens and subsequently used for Agrobacterium-mediated transformation of tobacco plants. Transgenic plants were selected on media containing kanamycin, and the expression of the transgene was verified through the fluorescence of green fluorescent protein. Microcallus formation and shoot development on selective media confirmed kanamycin resistance and the successful integration of the transgene. After phenotypic selection, genomic DNA was extracted from transgenic plants and analyzed by PCR (Polymerase Chain Reaction) using primers specific to the OMP25 gene. Positive PCR results validated the successful integration of the OMP25 gene into the plant genome. Gene expression was further confirmed at the RNA level through real-time quantitative PCR (qRT-PCR) and at the protein level via Western blot analysis. Future studies will evaluate immune responses in animal models. This approach demonstrates the potential for low-cost, effective vaccines to combat brucellosis, addressing critical economic and public health challenges.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"22"},"PeriodicalIF":2.7,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12041093/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144050499","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":"Integrated transcriptomic, transcriptional factors, and protein interaction reveal the regulatory mechanisms of flowering time in rice (Oryza sativa L.).","authors":"Amir Sohail, Liaqat Shah, Syed Mehar Ali Shah, Adil Abbas, Shahzad Ali, Manzoor","doi":"10.1007/s11248-025-00439-8","DOIUrl":"https://doi.org/10.1007/s11248-025-00439-8","url":null,"abstract":"<p><p>Appropriate flowering time is important for rice regional adaptation and optimum rice production, but little is known about the omics of heading date in rice. Here, we studied omics including transcriptome, proteome and transcriptional factors to identify regulatory genes related to flowering time. A total of 1402 differentially expressed genes (DEGs, 721 up-regulated and 681 down-regulated) were detected in wild and mutant. These transcripts are classified according to biological processes, cellular components, and molecular functions. Among these differentially expressed genes, many transcription factor genes demonstrated multiple regulatory pathways involved in flowering time. Gene expression analysis showed that Os03g0122600 (OsMADS50), Os08g0105000 (Ehd3), Os06g0275000 (Hd1) were expressed higher and Os06g0199500 (OsHAL3), Os06g0498800 (OsMFT1), Os08g0105000 (Ehd3), Os06g0157700 (Hd3a), and Os02g0731700 (Ghd2), were expressed lower in wild compared to mutant, which are the key genes that regulate the flowering in rice. In addition, Ghd7 interacted with Os10g30860 and Os12g08260 using yeast two-hybrid assay. We identified 28 potential Ghd7 transcriptional regulators using the transcription factor-centered yeast one hybrid (TF-Centered Y1H) assay. Taken together, this study developed a new set of genomic resources to identify and characterize genes, proteins, and motifs associated with flowering time.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"21"},"PeriodicalIF":2.7,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144041203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Abstracts of the 19th Transgenic Technology Meeting (TT2025) : Zurich, Switzerland, April 24-27, 2025.","authors":"","doi":"10.1007/s11248-025-00436-x","DOIUrl":"https://doi.org/10.1007/s11248-025-00436-x","url":null,"abstract":"","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 Suppl 1","pages":"19"},"PeriodicalIF":2.7,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144050077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced folate content in bioengineered lettuce stably expressed under field conditions.","authors":"Lídia N Queiroz, Tatiane R Monteiro, Ana C M M Gomes, Glaucia B Cabral, Francisco J L Aragão","doi":"10.1007/s11248-025-00440-1","DOIUrl":"https://doi.org/10.1007/s11248-025-00440-1","url":null,"abstract":"<p><p>Folate (vitamin B9) is an essential nutrient that plays a crucial role in various bodily functions. Its deficiency can lead to health issues, such as megaloblastic anemia, neural tube defects in the developing fetus and increased risk of cardiovascular diseases. Consequently, there is interest in increasing folate content in food crops by classical and molecular breeding. Since folate is a water-soluble vitamin that is sensitive to heat, we chose to manipulate its synthesis pathway in lettuce, which is often used for salads and sandwiches, without heat treatment, which makes it a good vehicle for making folate available to the population via biofortification. Transgenic lettuce plants were generated to express the GTP-cyclohydrolase I and aminodeoxychorismate synthase coding sequences from Arabidopsis thaliana. Plants were cultivated under greenhouse conditions and two field trials were carried out. Results have shown that transgenic lines presented up to 3.4 times more folate than the non-transgenic lettuce plants and 1.9 times more folate than spinach, considered one of the plants richest in folates. Advanced generations of homozygous plants were cultivated under field conditions for two years and the results showed that folate-biofortification was stable. A folate-biofortified lettuce serving would provide about 36 to 64% of the recommended daily intake. This technology is a foundation to produce folate-biofortified commercial varieties that can help to reduce hidden hunger, decreasing the number of cases of fetus malformations and other diseases.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"20"},"PeriodicalIF":2.7,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144015414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A method for detecting transgenic rapeseed using pollen collected by Apis mellifera L.","authors":"Rong Yuan, Miao Wang, Zhen Li, Meiyan Hong, Li Su, Gang Wu, Xinhua Zeng","doi":"10.1007/s11248-025-00438-9","DOIUrl":"10.1007/s11248-025-00438-9","url":null,"abstract":"<p><p>With the continuous expansion of the planting area of genetically modified (GM) crops, the demand for efficient and comprehensive monitoring systems is becoming increasingly urgent. To establish a method suitable for large-scale monitoring of genetically modified rapeseed planting, beehives were strategically deployed at specific locations around genetically modified rapeseed fields, and the TaqMan quantitative PCR (qPCR) method was used to detect and analyze the genetically modified components in the rapeseed pollen collected by bees. The results demonstrated that the average Ct values for the CaMV35S promoter, Bar gene, NPTII gene, and HPT gene in the pollen of each hive were 27.91, 29.58, 31.49, and 31.97, respectively. The average ΔCt values for these four genes in hive pollen from 100 to 200 m were - 0.35, 1.66, 2.58, and 5.06, respectively, which were significantly lower than those from 300 to 1100 m (2.85, 4.01, 6.66, and 5.63). The results of this study have demonstrated the feasibility of using pollen collected by bees for large-scale detection of genetically modified rapeseed plants. This early warning model for GM crop spread based on bee pollination provides an efficient and practical solution for monitoring and managing genetically modified crops.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"18"},"PeriodicalIF":2.7,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143789031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Shaker K⁺ channel NKT3A enhances potassium uptake and transport in tobacco (Nicotiana tabacum L.) seedlings under low potassium stress.","authors":"Haiying Xiang, Guang Yuan, Chuhan Shi, Li Xu, Jianduo Zhang, Qili Mi, Qian Gao, Wenwu Yang, Haitao Huang, Kunmiao Wang, Wanli Zeng, Yang Ning, Qian Wang","doi":"10.1007/s11248-024-00419-4","DOIUrl":"10.1007/s11248-024-00419-4","url":null,"abstract":"<p><p>One of the nutrients that is necessary for plant growth and development is potassium (K⁺). The uneven production and distribution of global potassium resources significantly challenge crop yields and quality. A moderate increase in the potassium content within plants can enhance both crop yield and quality. This study identifies the Shaker K⁺ channel NKT3A within the model crop, tobacco. The yeast heterologous expression system demonstrated its capability for K⁺ inward transportation. GUS staining and RT-qPCR analyses of the constructed promoter materials revealed NKT3A's activity during the tobacco seedling stage. Expression levels are higher in the leaf and stems, with low potassium levels inducing upregulation of its expression, also observed in roots. Gene editing technology was employed to construct overexpression and knockout mutants, with subsequent measurement of their phenotypes. Results indicate that NKT3A expression enhances facilitates potassium absorption and transport in tobacco seedlings under low potassium conditions. For the first time, this article identifies the Shaker potassium channel gene NKT3A, which functions as an inward rectifier K⁺ channel in tobacco. It elucidates the gene's role in regulating potassium distribution under low potassium conditions, thereby deepening our understanding of plant responses in such environments and offering a potential target for enhancing crop potassium use efficiency.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"17"},"PeriodicalIF":2.7,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143721573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plant-derived recombinant macromolecular PAP-IgG Fc as a novel prostate cancer vaccine candidate eliciting robust immune responses.","authors":"Yangjoo Kang, Deuk-Su Kim, Hyunjoo Hwang, Yerin Kim, Young-Jin Seo, Peter Hinterdorfer, Kisung Ko","doi":"10.1007/s11248-025-00433-0","DOIUrl":"10.1007/s11248-025-00433-0","url":null,"abstract":"<p><p>Prostatic acid phosphatase (PAP) is a specific protein that is highly expressed in prostate cancer. In this study, we constructed two recombinant PAP fusion genes: PAP fused to the immunoglobulin G (IgG) Fc fragment (designated PAP-Fc) and PAP-Fc fused to the endoplasmic reticulum retention sequence KDEL (designated PAP-FcK). Transgenic Nicotiana tabacum plants expressing these recombinant macromolecular proteins (MPs) were generated using Agrobacterium-mediated transformation, and the presence of both genes was confirmed through genomic PCR. Western blot analysis validated the expression of PAP-Fc and PAP-FcK MPs, which were successfully purified via protein A affinity chromatography. Size-exclusion high-performance liquid chromatography revealed dimeric peaks for PAP-Fc (PAP-Fc^P) and PAP-FcK (PAP-FcK^P). Bio-transmission electron microscopy demonstrated 'Y'-shaped protein particles resembling antibody structures. Moreover, PAP-Fc^P and PAP-FcK^P exhibited a high association rate with human FcγR and FcRn. Vaccination of mice with both PAP-Fc^P and PAP-FcK^P resulted in increased total IgG against PAP and enhanced activation of CD4⁺ T cells, comparable to mice immunized with PAP, which served as a positive control. These findings indicate that both plant-derived MPs can effectively induce adaptive immunity, positioning them as promising candidates for prostate cancer vaccines. Overall, plants expressing PAP-Fc and PAP-FcK represent a viable production system for antigenic macromolecule-based prostate cancer vaccines.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"16"},"PeriodicalIF":2.7,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143721572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}