Transgenic Research最新文献_第9页

Generation and characterization of cerebellar granule neurons specific knockout mice of Golli-MBP 小脑颗粒神经元特异性Golli-MBP基因敲除小鼠的产生和特征描述

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-29 DOI: 10.1007/s11248-024-00382-0

Haruko Miyazaki, Saki Nishioka, Tomoyuki Yamanaka, Manabu Abe, Yukio Imamura, Tomohiro Miyasaka, Nobuto Kakuda, Toshitaka Oohashi, Tomomi Shimogori, Kazuhiro Yamakawa, Masahito Ikawa, Nobuyuki Nukina

{"title":"Generation and characterization of cerebellar granule neurons specific knockout mice of Golli-MBP","authors":"Haruko Miyazaki, Saki Nishioka, Tomoyuki Yamanaka, Manabu Abe, Yukio Imamura, Tomohiro Miyasaka, Nobuto Kakuda, Toshitaka Oohashi, Tomomi Shimogori, Kazuhiro Yamakawa, Masahito Ikawa, Nobuyuki Nukina","doi":"10.1007/s11248-024-00382-0","DOIUrl":"https://doi.org/10.1007/s11248-024-00382-0","url":null,"abstract":"Golli–myelin basic proteins, encoded by the myelin basic protein gene, are widely expressed in neurons and oligodendrocytes in the central nervous system. Further, prior research has shown that Golli–myelin basic protein is necessary for myelination and neuronal maturation during central nervous system development. In this study, we established Golli–myelin basic protein-floxed mice to elucidate the cell-type-specific effects of Golli–myelin basic protein knockout through the generation of conditional knockout mice (Golli–myelin basic proteinsfl/fl; E3CreN), in which Golli–myelin basic proteins were specifically deleted in cerebellar granule neurons, where Golli–myelin basic proteins are expressed abundantly in wild-type mice. To investigate the role of Golli–myelin basic proteins in cerebellar granule neurons, we further performed histopathological analyses of these mice, with results indicating no morphological changes or degeneration of the major cellular components of the cerebellum. Furthermore, behavioral analysis showed that Golli–myelin basic proteinsfl/fl; E3CreN mice were healthy and did not display any abnormal behavior. These results suggest that the loss of Golli–myelin basic proteins in cerebellar granule neurons does not lead to cerebellar perturbations or behavioral abnormalities. This mouse model could therefore be employed to analyze the effect of Golli–myelin basic protein deletion in specific cell types of the central nervous system, such as other neuronal cells and oligodendrocytes, or in lymphocytes of the immune system.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"8 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140828151","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}

引用次数: 0

Regulatory landscape and public perception for gene-edited bananas in the Southeast Asian region 东南亚地区对基因编辑香蕉的监管情况和公众看法

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-10 DOI: 10.1007/s11248-024-00379-9

Nurzatil Sharleeza Mat Jalaluddin, Abdulah Al-Hadi Ahmad Fuaad, Rofina Yasmin Othman

{"title":"Regulatory landscape and public perception for gene-edited bananas in the Southeast Asian region","authors":"Nurzatil Sharleeza Mat Jalaluddin, Abdulah Al-Hadi Ahmad Fuaad, Rofina Yasmin Othman","doi":"10.1007/s11248-024-00379-9","DOIUrl":"https://doi.org/10.1007/s11248-024-00379-9","url":null,"abstract":"Banana is a premier fruit crop in many parts of the world especially Southeast Asia. The demand for banana has contributed to significant national income to primary banana producers in the SEA region such as the Philippines, Indonesia, Thailand, Vietnam, and Malaysia. However, the widely traded banana industry is plagued by numerous threats including pests and diseases, post-harvest issues and extreme climate vulnerability. To address these challenges, new breeding techniques such as gene editing have been explored for breeding programs to develop improved banana varieties. The first gene-edited non-browning banana has been deregulated in the Philippines recently, and more regulatory applications are expected to submit for approvals soon. Hence, it is timely to review the policy options for gene editing that have been adopted and discussed in the Southeast Asian countries and highlight the implications of differing regulatory approaches to gene editing for trading activities. Positive stakeholders’ perceptions and public acceptance are key factors in allowing the benefits of gene editing and thus appropriate outreach strategies are important to gain acceptance and avoid the “GMO stigma” that may be associated with gene-edited products.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"56 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140585818","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}

引用次数: 0

Prey-mediated effects of Mpp51Aa2-producing cotton on longevity and reproduction of Orius majusculus 产生 Mpp51Aa2 的棉花在猎物介导下对黄莺寿命和繁殖的影响

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-05 DOI: 10.1007/s11248-024-00378-w

Michael Meissle

{"title":"Prey-mediated effects of Mpp51Aa2-producing cotton on longevity and reproduction of Orius majusculus","authors":"Michael Meissle","doi":"10.1007/s11248-024-00378-w","DOIUrl":"https://doi.org/10.1007/s11248-024-00378-w","url":null,"abstract":"Genetically engineered (GE) cotton event MON 88702, producing Mpp51Aa2 (previously mCry51Aa2) from Bacillus thuringiensis (Bt), controls sucking pests, such as Lygus spp. (Hemiptera: Miridae) and thrips (Thysanoptera). Ingesting high doses of the insecticidal protein resulted in adverse effects on life table parameters of beneficial, predatory Orius spp. (Hemiptera: Anthocoridae). This triggered laboratory studies with more realistic food treatments, including different combinations of prey types with and without Bt protein to further characterize risks to this important group of non-target organisms. In this work, exclusive feeding of frozen spider mites (Tetranychus urticae, Acari: Tetranychidae) from Bt cotton confirmed adverse effects on longevity and fecundity of O. majusculus adults. Alternate feeding of Bt protein-containing spider mites and Bt-free Ephestia kuehniella (Lepidoptera: Pyralidae) eggs mitigated effects on longevity, but not on fecundity. When living larvae of Spodoptera littoralis (Lepidoptera: Noctuidae) from Bt cotton were fed to the predators, however, no effects on longevity and reproduction of female O. majusculus were observed, despite the fact that Bt protein concentrations in larvae were almost as high as concentrations in spider mites. When a diverse mix of prey species with various Bt protein concentrations is consumed in the field, it is unlikely that exposure of Orius spp. to Mpp51Aa2 is high enough to exert adverse effects on predator populations. MON 88702 cotton may thus be a valuable tool for integrated management of sucking pests.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"53 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140585789","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}

引用次数: 0

Membrane-based inverse-transition purification facilitates a rapid isolation of various spider-silk elastin-like polypeptide fusion proteins from extracts of transgenic tobacco 基于膜的反转纯化技术有助于从转基因烟草提取物中快速分离各种蜘蛛丝弹性蛋白样多肽融合蛋白

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-04 DOI: 10.1007/s11248-024-00375-z

H. M. Gruchow, P. Opdensteinen, J. F. Buyel

{"title":"Membrane-based inverse-transition purification facilitates a rapid isolation of various spider-silk elastin-like polypeptide fusion proteins from extracts of transgenic tobacco","authors":"H. M. Gruchow, P. Opdensteinen, J. F. Buyel","doi":"10.1007/s11248-024-00375-z","DOIUrl":"https://doi.org/10.1007/s11248-024-00375-z","url":null,"abstract":"Plants can produce complex pharmaceutical and technical proteins. Spider silk proteins are one example of the latter and can be used, for example, as compounds for high-performance textiles or wound dressings. If genetically fused to elastin-like polypeptides (ELPs), the silk proteins can be reversibly precipitated from clarified plant extracts at moderate temperatures of ~ 30 °C together with salt concentrations > 1.5 M, which simplifies purification and thus reduces costs. However, the technologies developed around this mechanism rely on a repeated cycling between soluble and aggregated state to remove plant host cell impurities, which increase process time and buffer consumption. Additionally, ELPs are difficult to detect using conventional staining methods, which hinders the analysis of unit operation performance and process development. Here, we have first developed a surface plasmon resonance (SPR) spectroscopy-based assay to quantity ELP fusion proteins. Then we tested different filters to prepare clarified plant extract with > 50% recovery of spider silk ELP fusion proteins. Finally, we established a membrane-based purification method that does not require cycling between soluble and aggregated ELP state but operates similar to an ultrafiltration/diafiltration device. Using a data-driven design of experiments (DoE) approach to characterize the system of reversible ELP precipitation we found that membranes with pore sizes up to 1.2 µm and concentrations of 2–3 M sodium chloride facilitate step a recovery close to 100% and purities of > 90%. The system can thus be useful for the purification of ELP-tagged proteins produced in plants and other hosts.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"25 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140602069","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}

引用次数: 0

The SpRY Cas9 variant release the PAM sequence constraint for genome editing in the model plant Physcomitrium patens SpRY Cas9 变体释放 PAM 序列约束，在模式植物 Physcomitrium patens 中进行基因组编辑

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-04 DOI: 10.1007/s11248-024-00381-1

Julie Calbry, Guillaume Goudounet, Florence Charlot, Anouchka Guyon-Debast, Pierre-François Perroud, Fabien Nogué

{"title":"The SpRY Cas9 variant release the PAM sequence constraint for genome editing in the model plant Physcomitrium patens","authors":"Julie Calbry, Guillaume Goudounet, Florence Charlot, Anouchka Guyon-Debast, Pierre-François Perroud, Fabien Nogué","doi":"10.1007/s11248-024-00381-1","DOIUrl":"https://doi.org/10.1007/s11248-024-00381-1","url":null,"abstract":"Genome editing via CRISPR/Cas has enabled targeted genetic modifications in various species, including plants. The requirement for specific protospacer-adjacent motifs (PAMs) near the target gene, as seen with Cas nucleases like SpCas9, limits its application. PAMless SpCas9 variants, designed with a relaxed PAM requirement, have widened targeting options. However, these so-call PAMless SpCas9 still show variation of editing efficiency depending on the PAM and their efficiency lags behind the native SpCas9. Here we assess the potential of a PAMless SpCas9 variant for genome editing in the model plant Physcomitrium patens. For this purpose, we developed a SpRYCas9i variant, where expression was optimized, and tested its editing efficiency using the APT as a reporter gene. We show that the near PAMless SpRYCas9i effectively recognizes specific PAMs in P. patens that are not or poorly recognized by the native SpCas9. Pattern of mutations found using the SpRYCas9i are similar to the ones found with the SpCas9 and we could not detect off-target activity for the sgRNAs tested in this study. These findings contribute to advancing versatile genome editing techniques in plants.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"26 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140586180","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}

引用次数: 0

Specific activity and utility of recombinant cellobiohydrolase II (Cel6A) produced in maize endosperm. 玉米胚乳中产生的重组纤维生物水解酶 II (Cel6A) 的特异性活性和实用性。

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-01 Epub Date: 2024-03-07 DOI: 10.1007/s11248-024-00376-y

Enio Duque Y Duque, Milena Aguirre, Nathan C Hood, Elizabeth E Hood

{"title":"Specific activity and utility of recombinant cellobiohydrolase II (Cel6A) produced in maize endosperm.","authors":"Enio Duque Y Duque, Milena Aguirre, Nathan C Hood, Elizabeth E Hood","doi":"10.1007/s11248-024-00376-y","DOIUrl":"10.1007/s11248-024-00376-y","url":null,"abstract":"Cellobiohydrolase II (CBH II) is an exo-glucanase that is part of a fungal mixture of enzymes from a wood-rot fungus, Trichoderma reesei. It is therefore difficult to purify and to establish a specific activity assay. The gene for this enzyme, driven by the rice Os glutelin promoter, was transformed into High II tissue culture competent corn, and the enzyme accumulated in the endosperm of the seed. The transgenic line recovered from tissue culture was bred into male and female elite Stine inbred corn lines, stiff stalk 16083-025 (female) and Lancaster MSO411 (male), for future production in their hybrid. The enzyme increases its accumulation throughout its 6 generations of back crosses, 27-266-fold between T1 and T2, and 2-10-fold between T2 and T3 generations with lesser increases in T4-T6. The germplasm of the inbred lines replaces the tissue culture corn variety germplasm with each generation, with the ultimate goal of producing a high-yielding hybrid with the transgene. The CBH II enzyme was purified from T5 inbred male grain 10-fold to homogeneity with 47.5% recovery. The specific activity was determined to be 1.544 units per µg protein. The corn-derived CBH II works in biopolishing of cotton by removing surface fibers to improve dyeability and increasing glucose from corn flour for increasing ethanol yield from starch-based first-generation processes.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":" ","pages":"47-57"},"PeriodicalIF":3.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140050437","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}

引用次数: 0

Establishment of a hydrodynamic delivery system in ducks. 建立鸭子水动力输送系统。

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-01 Epub Date: 2024-03-09 DOI: 10.1007/s11248-024-00377-x

Zhanji Zhao, Jiabing Zhu, Lijian Zhou, Nan Sun, Kaile Chang, Xiaoyue Hu, Yuting Hu, Mingzhi Ren, Yan Cheng, Derong Xu, Hongbo Xin, Chunbo Zhang

{"title":"Establishment of a hydrodynamic delivery system in ducks.","authors":"Zhanji Zhao, Jiabing Zhu, Lijian Zhou, Nan Sun, Kaile Chang, Xiaoyue Hu, Yuting Hu, Mingzhi Ren, Yan Cheng, Derong Xu, Hongbo Xin, Chunbo Zhang","doi":"10.1007/s11248-024-00377-x","DOIUrl":"10.1007/s11248-024-00377-x","url":null,"abstract":"Chronic hepatitis B virus (HBV) poses a significant global health challenge as it can lead to acute or chronic liver disease and hepatocellular carcinoma (HCC). To establish a safety experimental model, a homolog of HBV-duck HBV (DHBV) is often used for HBV research. Hydrodynamic-based gene delivery (HGD) is an efficient method to introduce exogenous genes into the liver, making it suitable for basic research. In this study, a duck HGD system was first constructed by injecting the reporter plasmid pLIVE-SEAP via the ankle vein. The highest expression of SEAP occurred when ducks were injected with 5 µg/mL plasmid pLIVE-SEAP in 10% bodyweight volume of physiological saline for 6 s. To verify the distribution and expression of exogenous genes in multiple tissues, the relative level of foreign gene DNA and β-galactosidase staining of LacZ were evaluated, which showed the plasmids and their products were located mainly in the liver. Additionally, β-galactosidase staining and fluorescence imaging indicated the delivered exogenous genes could be expressed in a short time. Further, the application of the duck HGD model on DHBV treatment was investigated by transferring representative anti-HBV genes IFNα and IFNγ into DHBV-infected ducks. Delivery of plasmids expressing IFNα and IFNγ inhibited DHBV infection and we established a novel efficient HGD method in ducks, which could be useful for drug screening of new genes, mRNAs and proteins for anti-HBV treatment.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":" ","pages":"35-46"},"PeriodicalIF":3.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140068757","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}

引用次数: 0

Multiplexed gene editing in citrus by using a multi-intron containing Cas9 gene. 使用含有多内含子的 Cas9 基因对柑橘进行多重基因编辑。

IF 3 3区生物学

Transgenic Research Pub Date : 2024-04-01 Epub Date: 2024-04-02 DOI: 10.1007/s11248-024-00380-2

Poulami Sarkar, Jorge Santiago Vazquez, Mingxi Zhou, Amit Levy, Zhonglin Mou, Vladimir Orbović

引用次数: 0

Efficiency of the alpha-hairpinin SmAMP-X gene promoter from Stellaria media plant depends on selection of transgenic approach 贝壳蕨类植物中α-发素 SmAMP-X 基因启动子的效率取决于转基因方法的选择

IF 3 3区生物学

Transgenic Research Pub Date : 2023-12-10 DOI: 10.1007/s11248-023-00374-6

Lyubov A. Ivanova, Roman A. Komakhin

{"title":"Efficiency of the alpha-hairpinin SmAMP-X gene promoter from Stellaria media plant depends on selection of transgenic approach","authors":"Lyubov A. Ivanova, Roman A. Komakhin","doi":"10.1007/s11248-023-00374-6","DOIUrl":"https://doi.org/10.1007/s11248-023-00374-6","url":null,"abstract":"The antimicrobial activity of the alpha-HAIRPININ ANTIMICROBIAL PEPTIDE X (SmAMP-X gene, GenBank acc. No. HG423454.1) from Stellaria media plant has been shown in vitro. Here, we isolated the SmAMP-X gene promoter and found two genomic sequences for the promoter (designated pro-SmAMP-X and pro-SmAMP-X-Ψ2) with 83% identity in their core and proximal regions. We found that the abilities of these promoters to express the uidA reporter and the nptII selectable marker differ according to the structural organization of T-DNA in the binary vector used for plant transformation. Analysis of Agrobacterium-infiltrated Nicotiana benthamiana leaves, transgenic Arabidopsis thaliana lines, and transgenic Solanum tuberosum plants revealed that both promoters in the pCambia1381Z and pCambia2301 binary vectors generate 42–100% of the ß-glucuronidase (GUS) activity generated by the CaMV35S promoter. According to 5’-RACE (rapid amplification of cDNA ends) analysis, both plant promoters are influenced by the CaMV35S enhancer used to express selectable markers in the T-DNA region of pCambia1381Z and pCambia2301. The exclusion of CaMV35S enhancer from the T-DNA region significantly reduces the efficiency of pro-SmAMP-X-Ψ2 promoter for GUS production. Both promoters in the pCambia2300 vector without CaMV35S enhancer in the T-DNA region weakly express the nptII selectable marker in different tissues of transgenic N. tabacum plants and enable selection of transgenic cells in media with a high concentration of kanamycin. Overall, promoter sequences must be functionally validated in binary vectors lacking CaMV35S enhancer.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"231 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2023-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138563153","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}

引用次数: 0

Efficient transformation and regeneration of transgenic plants in commercial cultivars of Citrus aurantifolia and Citrus sinensis. 枳壳和中华枳商业栽培品种中转基因植物的高效转化和再生。

IF 3 3区生物学

Transgenic Research Pub Date : 2023-12-01 Epub Date: 2023-09-13 DOI: 10.1007/s11248-023-00367-5

Sweta Singh, Zeba Tarannum, Sunil Kokane, Dilip K Ghosh, Ashwani K Sharma, Harsh Chauhan

{"title":"Efficient transformation and regeneration of transgenic plants in commercial cultivars of Citrus aurantifolia and Citrus sinensis.","authors":"Sweta Singh, Zeba Tarannum, Sunil Kokane, Dilip K Ghosh, Ashwani K Sharma, Harsh Chauhan","doi":"10.1007/s11248-023-00367-5","DOIUrl":"10.1007/s11248-023-00367-5","url":null,"abstract":"Citrus is one of the major horticultural crops with high economic and nutraceutical value. Despite the fact that conventional research has developed numerous improved varieties, citriculture is still susceptible to various stresses and requires innovative solutions such as genetic engineering. Among all the currently available modern approaches, Agrobacterium-mediated transformation is the most efficient method for introducing desired traits in citrus. However, being a non-host for Agrobacterium, various citrus species, including Citrus aurantifolia and Citrus sinensis, are recalcitrant to this method. The available reports on Agrobacterium-mediated transformation of commercial citrus cultivars show very low transformation efficiency with poor recovery rates of whole transgenic plantlets. Here, we provide an efficient and reliable procedure of Agrobacterium-mediated transformation for both C. aurantifolia and C. sinensis. This protocol depends on providing callus-inducing treatment to explants before and during Agrobacterium co-cultivation, using optimum conditions for shoot regeneration and modifying in-vitro micrografting protocol to combat the loss of transgenic lines. As transgenic citrus shoots are difficult to root, we also developed the ideal conditions for their rooting. Using this protocol, the whole transgenic plantlets of C. aurantifolia and C. sinensis can be developed in about ~ 4 months, with transformation efficiency of 30% and 22% for the respective species.","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":" ","pages":"523-536"},"PeriodicalIF":3.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10222383","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}

引用次数: 0