Plant Biotechnology Reports最新文献_第2页

Genome-wide identification and expression analysis of the SOD gene family under biotic and abiotic stresses in sweet orange 甜橙生物和非生物胁迫下 SOD 基因家族的全基因组鉴定和表达分析

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-07-10 DOI: 10.1007/s11816-024-00913-5

Xulin Li, Ke Wen, Tuo Yin, Chaoying Chen, Ling Zhu, Xiuyao Yang, Yinqiang Zi, Ke Zhao, Jiaming Zhang, Hanyao Zhang

{"title":"Genome-wide identification and expression analysis of the SOD gene family under biotic and abiotic stresses in sweet orange","authors":"Xulin Li, Ke Wen, Tuo Yin, Chaoying Chen, Ling Zhu, Xiuyao Yang, Yinqiang Zi, Ke Zhao, Jiaming Zhang, Hanyao Zhang","doi":"10.1007/s11816-024-00913-5","DOIUrl":"https://doi.org/10.1007/s11816-024-00913-5","url":null,"abstract":"Superoxide dismutase (SOD) protects plants from biotic and abiotic stress-induced reactive oxygen species toxicity and is extensively involved in plant growth and development. As the most widely cultivated and productive citrus fruit in the world, sweet oranges are susceptible to biotic and abiotic stresses during growth, affecting their yield and quality. However, the SOD gene family has not been identified in sweet oranges. Therefore, in this study, the sweet orange SOD gene family was systematically identified using bioinformatics methods, and a total of 15 sweet orange SOD (CsSOD) genes were identified and categorized into three subfamilies, Cu/Zn–SOD, Fe–SOD, and Mn–SOD, based on the results of the phylogenetic tree. Further analysis of gene structure and conserved motifs revealed that the motifs and exon and intron structures of CsSOD genes in the same subfamily were relatively identical, with only minor differences. In addition, we predicted hormone-related, light-response-related, and defense-related cis-acting elements in the promoters 2 kb upstream of the CsSOD genes. Transcriptome data analysis revealed that SOD genes were expressed under both abiotic and abiotic stresses, and the expression levels of some of the genes varied significantly. This study provides a basis for further understanding the biologic properties and functions of the SOD gene family in sweet oranges and provides a vital foundation for the study of sweet oranges under biotic and abiotic stresses.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"16 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141574154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Exogenous methyl jasmonate mediates tolerance of heat stress in Korean fir (Abies koreana) 外源茉莉酸甲酯介导韩国冷杉（Abies koreana）对热胁迫的耐受性

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-06-24 DOI: 10.1007/s11816-024-00912-6

Da Young Lee, Da Young Park, Hyeong Cheol Park

{"title":"Exogenous methyl jasmonate mediates tolerance of heat stress in Korean fir (Abies koreana)","authors":"Da Young Lee, Da Young Park, Hyeong Cheol Park","doi":"10.1007/s11816-024-00912-6","DOIUrl":"https://doi.org/10.1007/s11816-024-00912-6","url":null,"abstract":"Heat stress is a major environmental stress that affects the growth and development of plants. Korean fir (Abies koreana), a rare species endemic to South Korea, is sensitive to global climate change. The effect of exogenous methyl jasmonate (MeJA) on heat stress tolerance was, therefore, investigated in this species. During heat stress, the expression levels of eight genes (AkNAC19, AkMPK6, AkERF4, AkEFP, AkNAC2, AkbHLH, AkHSP17.6, and AkMYB123) were assessed in needles of A. koreana following treatment with 0, 0.1, 1.0, or 2.0 mM MeJA. Optimal upregulation of expression of most genes was observed 24 h post-treatment with 2.0 mM MeJA. Similar results were obtained when gene expression was analyzed 1, 2, 4, and 8 days post-treatment with 2.0 mM MeJA. Under heat stress conditions, plants treated with 2.0 mM MeJA initially showed a rapid decline in electrolyte leakage and higher chlorophyll content after 28 days of heat stress; however, opposite trends were observed in untreated plants, indicating that MeJA mediated tolerance to heat stress. Higher levels of expression of AkERF4, AkNAC2, and AkHSP17.6 were observed in MeJA-treated needles than in untreated needles, indicating these genes were strongly associated with MeJA-mediated heat tolerance. Therefore, these results suggest that the ability of Korean fir to tolerate abiotic stress is associated with endogenous MeJA synthesis or signaling, and identifies AkERF4, AkNAC2, and AkHSP17.6 as potential candidates for genes involved in the stress-tolerance mechanism.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"65 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141509101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Green biotherapeutics: overcoming challenges in plant-based expression platforms 绿色生物疗法：克服基于植物的表达平台所面临的挑战

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-06-21 DOI: 10.1007/s11816-024-00910-8

Rishabh Rajkumar Jadhav, Deepa Khare

{"title":"Green biotherapeutics: overcoming challenges in plant-based expression platforms","authors":"Rishabh Rajkumar Jadhav, Deepa Khare","doi":"10.1007/s11816-024-00910-8","DOIUrl":"https://doi.org/10.1007/s11816-024-00910-8","url":null,"abstract":"Plant-based expression platforms offer a promising avenue to produce biotherapeutics due to their scalability, cost-effectiveness, and potential for complex protein expression. However, the widespread adoption of these platforms faces significant challenges. This review provides an overview of the current landscape of plant-based expression systems, highlighting their advantages and limitations. Regulatory hurdles, concerns about product purity and consistency, technical limitations, and economic considerations are discussed as major obstacles to the utilization of plant-based platforms. Strategies for overcoming these challenges, including advancements in genetic engineering, optimization of protein expression, post-translational modification and quality, and implementation of stringent quality control measures, are explored. Case studies and success stories illustrate the feasibility and potential of plant-based expression platforms for commercial production. Future perspectives and opportunities for collaboration between academia, industry, and regulatory agencies are also discussed. Ultimately, this review aims to provide insights into the potential of plant-based expression platforms and the path forward to harness their full potential in the production of green biotherapeutics.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"30 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141509102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Nitric oxide mediated growth enhancement of tomato under salinity stress 一氧化氮介导的番茄在盐度胁迫下的生长促进作用

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-06-03 DOI: 10.1007/s11816-024-00909-1

Watheq Natiq Jumaah, Rizwana Begum Syed Nabi, Nkulu Kabange Rolly, Teferi Alem Adamu, Rupesh Tayade, Nay Chi Aye, Adil Hussain, Byung-Wook Yun

{"title":"Nitric oxide mediated growth enhancement of tomato under salinity stress","authors":"Watheq Natiq Jumaah, Rizwana Begum Syed Nabi, Nkulu Kabange Rolly, Teferi Alem Adamu, Rupesh Tayade, Nay Chi Aye, Adil Hussain, Byung-Wook Yun","doi":"10.1007/s11816-024-00909-1","DOIUrl":"https://doi.org/10.1007/s11816-024-00909-1","url":null,"abstract":"Salinity is a major problem for agricultural production throughout the world significantly limiting crop production. Here, we evaluated the effect of different concentrations of nitric oxide (NO) growth and development of five different tomato cultivars under salt stress induced by different concentrations of sodium chloride (NaCl). Results showed that germination was significantly reduced by the salt-stress treatments in a dose-dependent manner, where germination was significantly reduced by 75 mM NaCl but completely suppressed by 100 mM NaCl. Pre-treatment of seeds with 0.001 mM sodium nitroprusside (SNP) as a NO donor for 8 h not only accelerated the germination rate but also significantly improved the growth of seedlings under salt stress induced by 50 mM NaCl as compared to the salt-stressed plants not treated with SNP. Real-time PCR analysis showed that SNP treatment decreased the expression of antioxidant gene SlGRX1 after 6 and 12 h of the treatment but increased after 24 and 48 h. On the other hand, the expression of SlAPX1 was reduced at all time points, indicating a reactive oxygen species (ROS)-scavenging effect of the SNP treatment via GRX1 transcript accumulation. This suggests that NO plays a vital role in seed germination and early plant development. It is, therefore, concluded that exogenous NO treatment of tomato seeds can improve seed germination and plant growth under saline conditions.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"1 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141253485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Bioactivity assessment of high potency arachidin mixture from large-scale peanut hairy root culture medium 大规模花生毛根培养基中高效花生素混合物的生物活性评估

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-06-01 DOI: 10.1007/s11816-024-00907-3

Phadtraphorn Chayjarung, Kanjana Wongkrajang, Chanyanut Pankaew, Chonnikan Tothong, Wannakan Poonsap, Pakwuan Wongshaya, Siriwat Kucharoenphaibul, Anupan Kongbangkerd, Apinun Limmongkon

{"title":"Bioactivity assessment of high potency arachidin mixture from large-scale peanut hairy root culture medium","authors":"Phadtraphorn Chayjarung, Kanjana Wongkrajang, Chanyanut Pankaew, Chonnikan Tothong, Wannakan Poonsap, Pakwuan Wongshaya, Siriwat Kucharoenphaibul, Anupan Kongbangkerd, Apinun Limmongkon","doi":"10.1007/s11816-024-00907-3","DOIUrl":"https://doi.org/10.1007/s11816-024-00907-3","url":null,"abstract":"Peanut hairy root culture serves as a potent tool for producing valuable prenylated stilbenoids. A significant amount of chitosan (CHT), methyl jasmonate (MeJA), and methyl-β-cyclodextrin (CD) was used to elicit hairy roots in a bioreactor, which were later subjected to freeze-drying and extracted with acetone. The group Gr.4, obtained from the first-column chromatography and consisting of partially purified mixed arachidin, exhibited the highest antioxidant activity, measuring 3,067.53 ± 176.98 µmole Trolox/g crude extract. Moreover, when examining the antimicrobial activity, the partially purified Gr.4 showed the lowest minimum inhibition concentration (MIC) values against Staphylococcus aureus, Salmonella Typhimurium, Escherichia coli, and Candida albicans, with values of 31, 63, 125, and 78 µg/ml, respectively. Furthermore, the DNA nicking assay using the Fenton reaction demonstrated the DNA damage effect when plasmid DNA was exposed to concentrations of 25–100 µg/ml of partially purified Gr.4. Additionally, the scanning electron microscopy (SEM) results revealed irregularities and abnormalities in all tested microbial cells after treatment with 2xMIC of partially purified Gr.4. The enhanced activity exhibited by mixed arachidin compounds compared to the crude extract indicates a high potency of mixed arachidin. Moreover, the simplicity of isolation and purification suggests that the mixed arachidin compounds could be a preferable alternative for further applications as effective compounds rather than using them in individually purified forms.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"43 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141190636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Molecular identification and characterization of iris yellow spot virus (IYSV) and garlic virus X (GVX) infecting Allium species 感染鸢尾黄斑病毒（IYSV）和大蒜病毒 X（GVX）的分子鉴定和特征描述

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-05-31 DOI: 10.1007/s11816-024-00905-5

Muhammad Usman Shahid, Abid Riaz, Tahira Shafique, Mayasar I. Al-zaban, Diaa Abd El-Moneim, Sheikh Mansoor, Yong Suk Hung, Hyeon-Jin Sun

{"title":"Molecular identification and characterization of iris yellow spot virus (IYSV) and garlic virus X (GVX) infecting Allium species","authors":"Muhammad Usman Shahid, Abid Riaz, Tahira Shafique, Mayasar I. Al-zaban, Diaa Abd El-Moneim, Sheikh Mansoor, Yong Suk Hung, Hyeon-Jin Sun","doi":"10.1007/s11816-024-00905-5","DOIUrl":"https://doi.org/10.1007/s11816-024-00905-5","url":null,"abstract":"Allium species, such as Onion (Allium cepa L.) and garlic (Allium sativum L.), have been cultivated worldwide for centuries due to its enormous medicinal uses as well as for cooking practices. A large number of RNA viruses have been known to cause significant yield losses and also adversely affect the quality of Allium species. Among them, iris yellow spot virus (IYSV) and associated garlic virus X (GVX) from tospovirus family cause huge number of yield losses in onion and garlic crop. The disease due to IYSV and GVX was recorded in different locations on the basis of their characteristic symptoms. Experiments revealed that IYSV has the ability to transmit mechanically in a very efficient manner while the dispersal of GVX was only limited through its vector as compared to mechanical transmission. The disease incidence of IYSV on onion was reported about 80% while 2% of GVX was reported on garlic by mechanical transmission. Result revealed that IYSV is more viruliferous and has the ability to transmit more efficiently on onion as compared to GVX on garlic. PCR amplified IYSV samples of approximately 800 base pairs (bps) product using specific primer targeting coat protein (Cp) region, whereas no amplicon was detected for GVX infected samples. Phylogenetic analyses of 2 isolates RP13PK and RP27PK have shown 99.3% homogeneity with isolate DQ233469 as compared to remaining isolates. Therefore, the findings indicate that IYSV is a highly variable virus, undergoing rapid evolution in the region, necessitating vigilant monitoring and effective management. The outcomes imply that greater caution is required for controlling IYSV compared to GVX. Disease management strategies should be formulated with careful consideration of the swift evolution patterns exhibited by IYSV.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"50 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141190561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

New dimension in leaf stomatal behavior analysis: a robust method with machine learning approach 叶气孔行为分析的新维度：采用机器学习方法的稳健方法

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-05-09 DOI: 10.1007/s11816-024-00902-8

Ki-Bon Ku, Anh Tuan Le, Thanh Tuan Thai, Sheikh Mansoor, Piya Kittipadakul, Janejira Duangjit, Ho-Min Kang, San Su Min Oh, Ngo Hoang Phan, Yong Suk Chung

{"title":"New dimension in leaf stomatal behavior analysis: a robust method with machine learning approach","authors":"Ki-Bon Ku, Anh Tuan Le, Thanh Tuan Thai, Sheikh Mansoor, Piya Kittipadakul, Janejira Duangjit, Ho-Min Kang, San Su Min Oh, Ngo Hoang Phan, Yong Suk Chung","doi":"10.1007/s11816-024-00902-8","DOIUrl":"https://doi.org/10.1007/s11816-024-00902-8","url":null,"abstract":"Stomata are specialized pores that play a vital role in gas exchange and photosynthesis. Microscopic images are often used to assess stomatal characteristics in plants; however, this can be a challenging task. By utilizing Matterport’s Mask R-CNN implementation as the foundational model, fine-tuning was conducted on a dataset of 810 microscopic images of Hedyotis corymbosa leaves’ surfaces for automated stomatal pores detection. The outcomes were promising, with the model achieving a convergence of 98% mean average precision (mAP) for both detection and segmentation. The training loss and validation loss values converged around 0.18 and 0.37, respectively. Regression analyses demonstrated the statistical significance (p values ≤ 0.05) of predictor parameters. Notably, the tightest cluster of data points was observed in stomata pore area measurements, followed by width and length. This highlights the precision of the stomatal pore area in characterizing stomatal traits. Despite challenges posed by the original dataset’s low-resolution images and artifacts like dust, bubbles, and blurriness, our innovative utilization of the Mask R-CNN algorithm yielded commendable outcomes. This research introduces a robust approach for stomatal phenotyping with broad applications in plant biology and environmental studies.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"2 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140930741","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Phenotypic, biochemical and genetic diversity of pepper (Capsicum spp.) germplasm reflects selection for cultivar types and spatial distribution 辣椒（辣椒属）种质的表型、生化和遗传多样性反映了对栽培品种类型和空间分布的选择

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-05-02 DOI: 10.1007/s11816-024-00904-6

Sayed Azrah Indrabi, Ajaz Malik, Geetika Malik, Khursheed Hussain, Asif Shikari, Javid Iqbal Mir, Ji Eun Park, Anh Tuan Le, Sheikh Mansoor, Khalid Z. Masoodi

{"title":"Phenotypic, biochemical and genetic diversity of pepper (Capsicum spp.) germplasm reflects selection for cultivar types and spatial distribution","authors":"Sayed Azrah Indrabi, Ajaz Malik, Geetika Malik, Khursheed Hussain, Asif Shikari, Javid Iqbal Mir, Ji Eun Park, Anh Tuan Le, Sheikh Mansoor, Khalid Z. Masoodi","doi":"10.1007/s11816-024-00904-6","DOIUrl":"https://doi.org/10.1007/s11816-024-00904-6","url":null,"abstract":"Throughout the globe morphological, biochemical and genetic variability exists in chilli and is harnessed to achieve specific breeding objectives. In this study, chilli germplasm was characterized based on horticultural traits, biochemical quantification and simple sequence repeat (SSR) polymorphism for diversity estimation. A total of 36 SSR primers were utilised to study the genetic divergence among 48 genotypes of chilli collected from nine states of India. Among the 36 primers, sixteen amplified null alleles. A total of 41 alleles were detected with average 2.05 alleles per locus. The largest number of alleles (5) were obtained with marker CAMS-234. The polymorphic information content ranged from 0.06 to 0.72 with an average of 0.50. On the basis of SSR analysis, the UPGMA cluster classified 48 genotypes into three groups. There was significant variability in germplasm for all morpho-biochemical traits. Kashi Anmol (100.50 q/ha) expressed the highest yield. Highest vitamin C content at green stage was recorded in IC-561635 (187 mg/100 g) and the greatest capsaicin content (9547.90 µg/g) equivalent to pungency of 171,862.2 Scoville heat units (SHU) was recorded in Bhut Jolokia. Principal component analysis indicates that the first five principal components explain 74.63% per cent of the total variation. Additionally, analysis of molecular variance (AMOVA) showed that 1% of the total genetic variation occurred among the population and 99% genetic variation within the populations, whereas the pairwise Fst specified the moderate genetic variation ranging from 0.002 to 0.020. The present investigation has strengthened the knowledge of genetic worth of this germplasm for application in various genetic improvement programmes.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"15 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140886972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Influences of factors affecting the induction of high protoplast yield and callus induction in two different carnations 影响两种不同康乃馨原生质体产量和胼胝体诱导的因素的影响

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-04-29 DOI: 10.1007/s11816-024-00903-7

Oluwaseun Suleimon Adedeji, Aung Htay Naing, Jova Riza Campol, Kyeung Il Park, Chang Kil Kim

{"title":"Influences of factors affecting the induction of high protoplast yield and callus induction in two different carnations","authors":"Oluwaseun Suleimon Adedeji, Aung Htay Naing, Jova Riza Campol, Kyeung Il Park, Chang Kil Kim","doi":"10.1007/s11816-024-00903-7","DOIUrl":"https://doi.org/10.1007/s11816-024-00903-7","url":null,"abstract":"This study aims to establish an efficient protocol for protoplast isolation, cell division, and callus induction in two carnation cultivars, Chabaud and Giant Chabaud, by examining the influence of different types of plant tissue, enzyme concentrations, incubation times, cotyledon ages, and medium compositions. Our results indicate that protoplast yield varies significantly between different plant tissues, with true leaves offering the highest yield and viability, especially under a 0.1% driselase concentration and a 6-h incubation time. We observed that increasing the driselase concentration to 1.0% significantly reduced protoplast yields in all tissues tested. In addition, the age of the cotyledons notably affected protoplast yield, with younger cotyledons providing higher yields. The Murashige and Skoog medium supplemented with 1 mg/L zeatin and 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) proved to be the most effective for promoting cell division and colony formation from protoplasts derived from cotyledons and true leaves. The study also found that plant growth regulators (PGRs) significantly influence callus proliferation, with differences observed between protoplast sources.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"77 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140812768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Optimization of CRISPR/Cas9 ribonucleoprotein delivery into cabbage protoplasts for efficient DNA-free gene editing 优化 CRISPR/Cas9 核糖核蛋白向卷心菜原生质体的输送，实现高效的无 DNA 基因编辑

IF 2.4 4区生物学

Plant Biotechnology Reports Pub Date : 2024-04-10 DOI: 10.1007/s11816-024-00901-9

Sora Lee, Su Hyun Park, Yu Jeong Jeong, Soyoung Kim, Bo Ryeong Kim, Bo-Keun Ha, Chaein Na, Jiyoung Lee, Jae Cheol Jeong, Cha Young Kim

{"title":"Optimization of CRISPR/Cas9 ribonucleoprotein delivery into cabbage protoplasts for efficient DNA-free gene editing","authors":"Sora Lee, Su Hyun Park, Yu Jeong Jeong, Soyoung Kim, Bo Ryeong Kim, Bo-Keun Ha, Chaein Na, Jiyoung Lee, Jae Cheol Jeong, Cha Young Kim","doi":"10.1007/s11816-024-00901-9","DOIUrl":"https://doi.org/10.1007/s11816-024-00901-9","url":null,"abstract":"The CRISPR/Cas9-based gene editing system for the direct delivery of pre-assembled Cas9 ribonucleoproteins (RNPs), consisting of a Cas9 nuclease and a single guide RNA (sgRNA), into plant protoplasts enables DNA-free gene editing without introducing foreign gene into plants. Here, we described the optimization of CRISPR/Cas9 RNPs delivery into cabbage protoplasts for efficient DNA-free gene editing. We determined the insertion and deletion (indel) frequency of BoMYBL2-1, a negative regulatory gene for anthocyanin biosynthesis in cabbage (Brassica oleracea var. capitata). We optimized the molar ratio of Cas9 to sgRNA and the incubation time of RNP–protoplast transfection to enhance the indel frequency under various conditions. Based on the BoMYBL2-1 nucleotide sequences, we designed nine sgRNAs to target BoMYBL2-1. Our in vitro digestion assay showed that all sgRNAs were able to cleave the targeted fragment. When the sgRNA and Cas9 proteins were subsequently transfected into protoplasts isolated from cabbage cotyledons, the deep sequencing results showed that the indel frequency of sgRNAs in BoMYBL2-1 was the highest (7.4%) with sgRNA3. We compared various molar ratios of Cas9 and sgRNA and incubation times of RNP–protoplast transfection to optimize transfection and ensure high indel frequency. The highest frequency was observed when the Cas9:sgRNA ratio was 1:10. Furthermore, when the incubation time for RNP–protoplast transfection was 1 min and 3 min, the indel frequency was higher than 25%. Altogether, these results provide valuable information on the optimized conditions for high-efficiency gene editing using CRISPR/Cas9 RNP delivery into cabbage protoplasts.","PeriodicalId":20216,"journal":{"name":"Plant Biotechnology Reports","volume":"148 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140588837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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