In Vitro Cellular & Developmental Biology - Plant最新文献

{"title":"The alternative approaches to anthocyanin production by callus culture of Vaccinium arctostaphylos L. and the ultrastructure of anthocyanin-producing callus","authors":"Havva Karahan, Elif Onan, Hatice Çölgeçen","doi":"10.1007/s11627-024-10443-y","DOIUrl":"https://doi.org/10.1007/s11627-024-10443-y","url":null,"abstract":"<p>In this study, the aim was to try to increase the anthocyanin-producing in callus and to investigate the ultrastructure of the developing callus cells of leaf, apical meristem, and node explants of <i>Vaccinium arctostaphylos</i> L. To generate callus, sterilized explants were seeded in Woody Plant Medium (WPM) containing different concentrations of indoleacetic acid or zeatin. The most calluses containing anthocyanin were obtained from the apical meristem explant cultured on medium containing 2.0 mg L⁻¹ indoleacetic acid. The medium containing 2.0 mg L⁻¹ indoleacetic acid was modified with different sucrose concentrations and were prepared to increase anthocyanin production and callus biomass. The best callus production was obtained with apical meristem explants cultured on medium containing 50.0 g L⁻¹ sucrose and was observed as a pink color callus when examined under a fluorescence microscope with 525-nm green and 610-nm red wavelengths. The callus had autofluorescent. The cell ultrastructures of the pink- and yellow-colored callus growing in medium containing 50.0 g L⁻¹ sucrose were examined using a transmission electron microscope (TEM) after the preparation process. Cells that developed from yellow callus and were called type 1 had an elliptical or long, angular shape. Type 1 cells had a large vacuole. There were electron-dense vesicles around the vacuoles attached to the inner surface of the tonoplast, and anthocyanic vacuolar inclusion–like inclusions were seen in the vacuole. The cytoplasm and organelles were trapped between the vacuole and the cell wall. It was observed that cells developed from pink color callus (called type 2) often had an elliptical shape. Similar to type 1 cells, it has a large vacuole; its cytoplasm is trapped between the cell wall and the vacuole with very few intercellular spaces. Some of the vesicles had an anthocyanic vacuolar inclusion appearance within the vacuole. A macroautophagy-like fusion was placed between the cytoplasmic vesicles and the vacuole. These observations showed that the anthocyanin-producing cells of the <i>V. arctostaphylos</i> L. transported anthocyanins to the vacuole using the vesicular transport method. The current research is the first study to fill the gap in the literature, so it is extremely important as a source for future studies.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"15 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142212864","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":"Factors influencing the in vitro growth and maintenance of several Vietnamese accessions of the duckweed Lemna aequinoctialis (Welw.)","authors":"Hoang Thi Nhu Phuong, Tran Nguyen Kim Ngan, Tran Thi Nhung","doi":"10.1007/s11627-024-10451-y","DOIUrl":"https://doi.org/10.1007/s11627-024-10451-y","url":null,"abstract":"<p>This study investigates the influence of various parameters on the efficiency of cultivating and preserving <i>Lemna aequinoctialis</i> duckweed samples <i>in vitro</i>, with the aim of minimizing time, labor, and storage expenses. The method addresses the issue of limited space and the risk of sample loss by storing multiple duplicates (more than one tube or flask per sample) in a space-efficient and cost-effective container. Due to the practical implications and capacity to decrease operating expenses, our results are helpful for large-scale implementation. Using a single frond resulted in a longer preservation effect compared to sample densities of three and five fronds. Initial samples can be cultured in nutrient solution with or without sugar, stored in tap water or nutrient solution, kept at a stable temperature of 25 ± 2°C or room temperature. The test tubes with duckweed were not covered, or covered by black paper up to the solution surface or higher. The outcomes demonstrated that all three duckweed samples (HNP_005, HNP_026, and HNP_031), whether grown in SSM or SFM media, were best preserved if the tubes were covered by black paper higher than the solution surface (CH) and can be kept at room temperature for at least 9 mo.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"16 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142212862","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":"Elimination of sugarcane mosaic virus, sugarcane yellow leaf virus, and co-infections in sugarcane (Saccharum spp. hybrids) shoot tips via osmo- and cryo-therapy","authors":"Khethumusa H. Cele, Meenu Ghai, Sandra J. Snyman","doi":"10.1007/s11627-024-10449-6","DOIUrl":"https://doi.org/10.1007/s11627-024-10449-6","url":null,"abstract":"<p>Cryopreservation for sugarcane (<i>Saccharum</i> spp. hybrids) germplasm conservation is well established. Virus elimination using droplet-vitrification (D-V) and cryo- or osmo-therapy has only been recently reported for sugarcane mosaic virus (SCMV). In this study, exposing large (3 mm) <i>in vitro</i> shoot tips of cultivars N12, N19, N58, and NCo376 infected with sugarcane yellow leaf virus (SCYLV) and NCo376 co-infected with SCMV and SCYLV were tested for virus elimination using both of the above-mentioned techniques. Cryo-therapy involved the exposure of infected <i>in vitro</i> shoot tips to the D-V protocol followed by recording recovery and virus-free shoot tips 16 wk after treatment. Osmo-therapy, consisting of the same treatment as cryo-therapy without immersion in liquid nitrogen (LN), was included for comparative purposes. Cryo-therapy resulted in 100% of the recovered shoots being SCYLV-free in cultivars N19, N58, and NCo376 and 83% in N12 when compared with untreated material. Osmo-therapy showed 58% (N12), 91% (N19 and N58), and 100% (NCo376) of shoots being clear of SCYLV when compared with untreated <i>in vitro</i> control plants (0 to 8%). Both techniques reduced the regrowth levels of treated shoot tips (22 to 57% recovery) when compared with untreated controls (92 to 97%). A novel finding of the study was that NCo376 co-infected with SCMV and SCYLV showed 100% virus-free recovered shoots after cryo-therapy and 92 to 100% of healthy shoots after osmo-therapy, compared with controls, which had 17 to 42% virus-free shoots. Plants from all cultivars that were re-tested 4 mo after hardening maintained their virus-free status. The described techniques for virus eradication offer a promising solution for the provision of clean vegetative planting propagules and safer germplasm exchange.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"7 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142212899","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":"Micropropagation and genetic transformation of Byblis liniflora","authors":"Alberto Coronado-Martín, Constanza Martin-Vásquez, Marybel Jáquez, Abdellatif Bahaji, Alejandro Atarés","doi":"10.1007/s11627-024-10448-7","DOIUrl":"https://doi.org/10.1007/s11627-024-10448-7","url":null,"abstract":"<p><i>Byblis</i>, a small genus of carnivorous plants predominantly found in Australia, is characterized by its passive trapping mechanism and unique floral features. The chemical composition of <i>Byblis</i>, including identified phenylethanoid glycosides, particularly acteoside, highlights its pharmacological potential with various biological activities. <i>In vitro</i> culture techniques have been established for propagation, with micropropagation protocols developed for different <i>Byblis</i> species. However, information on genetic transformation, vital for trait modification and enhanced pharmacological interest, remains limited. This study focuses on optimizing micropropagation, adventitious regeneration, and genetic transformation methods for <i>Byblis liniflora</i>. Adventitious regeneration rates were highest in medium with reduced Murashige and Skoog salts (MS/10) and sucrose (3 gL⁻¹) concentrations. Zeatin supplementation (1 mgL⁻¹) further improved regeneration rates and bud development with 100% of regenerated root explants and 8.8 shoots per explant. Liquid MB3 medium supplemented with indole-3-acetic acid (IAA) 5 mgL⁻¹ facilitated efficient rooting and acclimatization. The establishment of an efficient <i>Rhizobium</i>-mediated genetic transformation method yielded transgenic plants expressing green fluorescent protein (GFP). Molecular analysis confirmed transgene integration, marking the first successful genetic transformation in the <i>Byblis</i> genus. These advancements pave the way for exploring gene function and enhancing pharmacological properties, thereby broadening our understanding and utilization of carnivorous plants like <i>Byblis</i>.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"83 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141946761","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":"In vitro regeneration of cotton (Gossypium hirsutum L.) cultivar KC3 with controlled phenolic secretion by using Kappaphycus alvarezii sulfated polysaccharide extract and plant growth regulators","authors":"Packiaraj Gurusaravanan, Sathasivam Vinoth, Rajkumar Vasanthkumar, Muthukrishnan Arun, Muthukrishnan Saradhadevi, Subiramani Sivakumar, Narayanasamy Jayabalan","doi":"10.1007/s11627-024-10444-x","DOIUrl":"https://doi.org/10.1007/s11627-024-10444-x","url":null,"abstract":"<p>An efficient and reproducible <i>in vitro</i> regeneration protocol was developed for elite cotton cultivar KC3 by using plant growth regulators (PGRs) in combination with seaweed polysaccharide (SP) extracts. The existence of polysaccharide in seaweed extract was confirmed by Fourier transform infrared spectroscopy (FT-IR) and carbon-13 (¹³C) nuclear magnetic resonance (NMR) spectroscopy analysis. The extracted SP extract efficacy was tested for <i>in vitro</i> plant regeneration. The maximum callus frequency (89.4%) was obtained from hypocotyl explant in the Murashige and Skoog (MS) medium supplemented with 4.0% glucose, 1.5 mg L⁻¹ thidiazuron (TDZ), 0.6 mg L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), and 30.0 mg L⁻¹ SP. Remarkably, PGR- and SP-fortified medium inhibits the phenolic excretion from the explants. The well-developed yellow green friable texture of callus was transferred to shoot initiation medium. MS medium fortified with 4.0% glucose, 2.0 mg L⁻¹ 6-(γ,γ-dimethylallylamino) purine (2iP), 1.0 mg L⁻¹ kinetin (KIN), 1.0 mg L⁻¹ 6-benzylaminopurine (BA), and 40.0 mg L⁻¹ SP has shown maximum response (85.2%), and it produced 9.5 shoots per callus. The elongated shoots were cultured on root induction medium which consists of Murashige and Skoog (MS) salts with indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), and SP. The results revealed that the maximum number of roots (12.9 per shoot) with 8.6 cm in length was achieved on MS medium supplemented with 0.6 mg L⁻¹ IBA, combined with 30.0 mg L⁻¹ SP. Therefore, modified MS medium with natural bio-stimulant has more potential and is more reliable for <i>in vitro</i> regeneration of plants by neutralizing the effects of phenolic compounds secreted by the explants.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"43 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141867031","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":"Identification of methyl jasmonate-induced cardiac glycosides and related biosynthetic transcripts from callus culture of Calotropis gigantea using transcriptome and metabolite profiling","authors":"Pankaj Singh, Akansha Pandey, Carol Janis Bilung, Amar Jeet, Renu Nimoriya, Shiv Nandan, Sanjeev Kanojia, Dipak Kumar Mishra, Vineeta Tripathi","doi":"10.1007/s11627-024-10446-9","DOIUrl":"https://doi.org/10.1007/s11627-024-10446-9","url":null,"abstract":"<p>Cardiac glycosides (CGs) are well known for treating congestive heart failure, and several CGs like digoxin, digitoxin, and ouabain are marketed as drugs. In the present study, we have biosynthesized two CGs (CGCL520/227 and CGCL534/209) and elicited them up to 537- and 357-fold respectively in response to methyl jasmonate (MJ) treatment. For identification of the key enzyme involved in its biosynthesis, a comparative transcriptome sequencing of control and MJ elicited (75.0 mg L⁻¹ for 3 d) callus culture was done. A total of 17,898 transcripts were expressed across all samples. Annotated unigenes were functionally categorized based on gene ontology. A total of 7625 unigenes were significantly matched in the KEGG database involved in 151 different plant metabolism pathways. Upon digital expression analysis, 2924 MJ-responsive transcripts were identified, and among them 166 were unique for MJ-treated samples. A majority of upregulated transcripts were categorized under hydrolase activity, oxido-reductase activity, metabolic processes, and carbohydrate metabolic process. Based on their role in terpenoid, steroid, and cardenolide pathways, 295 putative unigenes representing 24 gene families involved in CG biosynthesis were identified. Expression analysis revealed that 12 transcripts involved in steroid and cardenolide biosynthetic pathways were upregulated in response to MJ. The highest expression was recorded for <i>squalene monooxygenase</i> (SMO) with 43-fold upregulation, followed by <i>sterol delta7 reductase</i> (DWF5) with 22.2-fold. <i>C-5 sterol desaturase</i> (STE1), <i>4-diphosphocytidyl-2-C-methyl-D-erythritolkinase/4diphosphocytidyl-2C-methyl-D-erythritol synthase</i> (CMK), <i>4-hydroxy-3-methylbut-2-enyl diphosphate reductase</i> (HDR), <i>acetyl-CoA C-acetyltransferase</i> (AACT), <i>mono-oxygenases</i> (MO), and <i>progesterone 5β-reductase</i> (PBR) showed high and significant expressions of 16.4-, 16.1-, 14.8-, 14.7-, 13.4-, and 11.3-fold, respectively. This study not only identifies MJ-responsive CGs and related transcripts involved in CG biosynthesis, but also provides scope for the development of biotechnological process for biosynthesis and enrichment of targeted CGs using identified rate-limiting key enzymes.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"34 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141867088","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":"Improvement of Stachys sieboldii (Miq.) growth by virus elimination of shoot apices cultivated on media free of plant growth regulators","authors":"Jizhi Jin, Fangyuan Zhou, Meng Yang, Wei Sheng, Yongbo Duan, Fenglan Zhao","doi":"10.1007/s11627-024-10440-1","DOIUrl":"https://doi.org/10.1007/s11627-024-10440-1","url":null,"abstract":"<p>Chinese artichoke (<i>Stachys sieboldii</i> [Miq.]) is a popular healthcare food owing to its high contents of stachyose. Viral infections have caused the severe degeneration of germplasm in this plant species. An efficient virus elimination and micropropagation system is required to produce Chinese artichoke plants that are free of viruses. A protocol for virus elimination in Chinese artichoke was established by comparing the potential for growth of the terminal buds using shoot apices as starting materials, in media with or without plant growth regulators (PGRs) and various concentrations of sucrose. Murashige and Skoog (MS) medium without any PGR performed better than that supplemented with PGRs as indicated by a higher rate of survival of shoot apices, higher plant height, more root numbers and adventitious buds, and increased biomass (<i>P</i> &lt; 0.05). An experiment to further optimize the sucrose concentration demonstrated that MS medium with 5.0% sucrose (w/v) efficiently induced the growth of both roots and shoots, thus, achieving the efficient micropropagation of Chinese artichoke within 4 wk. Compared with the mother plants in which viruses had not been eliminated, the virus-free plants had significantly higher numbers of tubers, increased yield, and a higher content of stachyose (<i>P</i> &lt; 0.05). Higher contents of endogenous hormones were detected in Chinese artichoke, which may explain the efficient micropropagation without the use of exogenous PGRs. This simple protocol enabled the production of virus-free Chinese artichoke to enhance the yield of tubers and high content of stachyose.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"145 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141503378","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":"Establishment of an efficient regeneration system and in vitro polyploid induction based on the bulblet centre in Lilium rosthornii Diels","authors":"Yongyao Fu, Xiaomeng Liang, Hong Zhang, Shiyuan Cheng, Anqin Li, Minjing Liao, Lang Tan, Liping Yang, Xiangying Qi","doi":"10.1007/s11627-024-10438-9","DOIUrl":"https://doi.org/10.1007/s11627-024-10438-9","url":null,"abstract":"<p>\u0000<i>Lilium rosthornii</i> Diels, commonly known as the Nanchuan lily in China, is a unique wild lily species with potential medicinal or ornamental value. The development and utilization are mainly restricted to its highly efficient regeneration and the scarcity of new germplasm. In this study, an efficient plant regeneration method for Nanchuan lily was established based on embryogenic callus induction, adventitious bud sprouting, and shoot proliferation and rooting. The results showed that Murashige and Skoog (MS) media supplemented with 0.5 mg L⁻¹ picloram (PIC) and 1.0 mg L⁻¹ 1-naphthaleneacetic acid (NAA) were suitable for embryogenic callus induction using outer bulblet scales (78.89% rate), and MS supplemented with 0.5 mg L⁻¹ 6-benzylaminopurine (6-BA), 1.0 mg L⁻¹ NAA, and 75.0 mg L⁻¹ ascorbic acid in darkness was optimal for embryogenic callus proliferation (2.62 coefficient). Moreover, callus inoculation in MS medium supplemented with 1.0 mg L⁻¹ thidiazuron (TDZ) and 0.1 mg L⁻¹ NAA significantly promoted shoot induction (3.16 coefficient) under relatively low light conditions. The shoot proliferation and rooting of MS plants treated with 0.1 mg L⁻¹ NAA and 0.5 mg L⁻¹ TDZ were greatest under light conditions at 93.33% and 98.89%, respectively. In addition, bulblet growth was induced by treatment with different combinations of colchicine at different concentrations for different durations. Focusing on the bulblet centre, 100% morphological changes were achieved for <i>in vitro</i> induction using 0.15% colchicine for 72 h of treatment. Compared with the control, three stable mutagenic plants with greater guard cells and reduced stomatal density were generated and were identified as chimaeras by flow cytometry analysis. These results provide support for improving tissue reproduction and breeding autotetraploid germplasm in Nanchuan lily.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"57 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141503380","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":"Tissue culture and high-efficiency transformation in an apomictic initial variety of Paspalum notatum Flüggé","authors":"Rafael Narancio, Daniel Isenegger, Rafael Reyno, German Spangenberg, Marco Dalla-Rizza","doi":"10.1007/s11627-024-10434-z","DOIUrl":"https://doi.org/10.1007/s11627-024-10434-z","url":null,"abstract":"<p><i>Paspalum notatum</i> Flüggé is a subtropical grass native from the Americas and one of the main components of the subtropical South American grasslands. Although <i>P. notatum</i> exhibits a high forage productivity and persistence, some limitations are observed in the species, such as forage quality due to high lignin content. The development of efficient tissue culture and plant transformation protocols in cultivars for forage production are necessary to embrace novel genetic improvement through transgenesis and genome editing. Efficient regeneration and transformation systems for many plant species can be hampered by genotype dependency, which can only be determined through extensive laboratory testing and evaluation. In this study, various factors extracted from protocols have been evaluated to establish efficient somatic embryogenesis, plant regeneration, and genetic transformation in apomictic initial variety INIA Sepé. Optimal somatic embryogenesis and regeneration from explant sources, such as mature seeds and meristematic tissue derived from <i>in vitro</i> propagated tillers, ranged between 79 and 88%. Microprojectile bombardment of two plasmids encoding a constitutively regulated reporter gene and <i>nptII</i> selectable marker expression constructs showed an average transformation efficiency of 10.3% from meristematic tissue as an explant source. Furthermore, the constitutive promoters, <i>35S</i> and <i>ZmUbi</i>, were tested in <i>GFP</i> transient expression assays to evaluate optimal cassettes for transgene(s) expression. Translation of a wheat codon–optimized <i>Cas9</i> sequence using <i>Cas9:2A:tuGFP</i> fusion protein was also tested in protoplasts. The information generated on promoter activity and Cas9 translation, and the establishment of an efficient transformation system, provided useful tools for future work in genetic engineering and genome editing in <i>P. notatum</i> cv. INIA Sepé.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"22 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141503379","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":"Mannitol and sorbitol concentration optimization for effective Epipactis flava Seidenf. in vitro slow growth storage","authors":"Julaluk Linjikao, Phithak Inthima, Apinun Limmongkon, Anupan Kongbangkerd","doi":"10.1007/s11627-024-10437-w","DOIUrl":"https://doi.org/10.1007/s11627-024-10437-w","url":null,"abstract":"<p><i>Epipactis flava</i> Seidenf., a Thai rheophytic orchid, is endangered due to habitat destruction and climate change. <i>In vitro</i> conservation serves as an effective technique for the <i>ex situ</i> preservation of orchid diversity. This study investigated the <i>in vitro</i> storage of <i>E</i>. <i>flava</i> seedlings under slow growth conditions using Murashige and Skoog (MS) medium supplemented with either sorbitol or mannitol individually at concentrations of 1.0%, 2.0%, 3.0%, and 4.0% (<i>w/v</i>) for 24 wk without subculture. The storage medium added with 3.0% (<i>w/v</i>) sucrose was used as the control. After 24 wk of storage, the storage medium containing 2.0% mannitol provided the highest survival rate (97.6%) while the highest leaf formation (70.8%) was achieved with 3.0% sucrose. The addition of 1.0 to 2.0% sorbitol, 1.0% mannitol, and 3.0 to 4.0% mannitol showed the highest shoot bud formation (100%). At the end of 24 wk, the plantlets were transferred to fresh medium for an 8-wk growth recovery. The plantlets derived from the storage medium with 3.0% mannitol exhibited the highest survival rate (73.5%). Conversely, the highest number of shoot buds (9.9 shoot buds per plantlet) was found on plantlets derived from storage medium with 1.0% sorbitol while the highest number of shoots (4.8 shoots per plantlet) was achieved in plantlets derived from 1.0% mannitol. Consequently, these results suggested that the storage medium with 3.0% mannitol is optimal for conserving plantlets for up to 24 wk, and all plantlets could successfully produce shoot buds and shoots during <i>in vitro</i> recovery. Hence, this strategy has potential in the conservation and utilization of this endangered species.</p>","PeriodicalId":13293,"journal":{"name":"In Vitro Cellular & Developmental Biology - Plant","volume":"41 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141252995","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}