Plant Cell, Tissue and Organ Culture最新文献

{"title":"Direct regeneration and genetic transformation studies in Hemidesmus indicus (L.) R. Br. (Indian Sarsaparilla)","authors":"Manjula Ranganatha, Ashwani Sharma, B.E. Rangaswamy, Shashi Kumar, Nagashree N. Rao","doi":"10.1007/s11240-024-02771-z","DOIUrl":"https://doi.org/10.1007/s11240-024-02771-z","url":null,"abstract":"<p><i>Hemidesmus indicus</i> L. R. Br. is a rare and <i>endangered</i> plant exhibits potent medicinal properties explored for the presence of flavoring compound in the roots namely 2-hydroxy-4- methoxybenzaldehyde (HMB) having applications in pharmaceutical and nutraceutical industries. The objective of the study is to improve the regeneration potential and optimize the genetic transformation in <i>Hemidesmus indicus.</i> Present study exhibits direct regeneration of <i>Hemidesmus indicus</i> through leaf and nodal explants with subsequent plant regeneration using Murashige and Skoog (MS) medium supplemented with various plant growth regulators (auxins, cytokinins, and gibberellic acid) TRIA and with adjuvant adenine sulphate,. The caulogenic response of 78.8%, 73.3% and 71.4% was observed when the leaf explant was inoculated on MS media containing 2.3 mgL^− 1 BAP + 0.2 mgL^− 1 2,4-D, 0.02 mgL^− 1 TRIA + 2 mgL^− 1 BAP and 1 mgL^− 1 KIN + 1 mgL^− 1 NAA respectively with creamish yellow nodular friable callus by 4 weeks. The initiation of shoot bud was observed within three days after inoculation of nodal explant on media supplemented with 1 mgL^− 1 BAP + 0.1 mgL^− 1 NAA, 1 mgL^− 1 BAP + 0.1 mgL^− 1 NAA + 40 mgL^− 1 AgNO₃, 1 mgL^− 1 BAP + 0.1 mgL^− 1 NAA + 40 mgL^− 1 AgNO₃ + 40 mgL^− 1 adenine sulphate respectively and incubated in the dark for 2 weeks. Shoot regeneration from the leaf explants was also observed within 4 weeks after inoculation in MS medium with 1 mgL^− 1 BAP + 0.1 mgL^− 1 NAA. In the present study, we also report the development of an effective and reproducible <i>Agrobacterium tumefaciens</i> mediated genetic transformation system in <i>Hemidesmus indicus</i>. Genetic transformation was achieved by inoculating in vitro calli of <i>Hemidesmus indicus</i> with the <i>A. tumefaciens</i> strain EHA105 harboring pCAMBIA1301 with 35 S CAMV promoter driving the expression of reporter β-glucuronidase gene (GUS) and hygromycin selection marker hptII. After co-cultivation, the calli were thoroughly washed and inoculated on MS basal medium with (2.3mgL^− 1 BAP + 0.2 mgL^− 1 2,4-D) along with cefotaxime (250 mgL^− 1) and hygromycin (30 mgL^− 1) and the co-cultivated callus cultures were maintained in dark at 25 ̊C ± 2 ̊C for 4 weeks. After two rounds of subculture, the putative transformed calli were assessed by performing histochemical GUS staining assay and genomic DNA PCR to detect the exogenous GUS gene sequence. The transformation efficiency was 26% with transformed calli tolerant to hygromycin (30 mg L⁻¹) and were positive to histochemical GUS staining assay and PCR. The results indicated a successful establishment of a reliable and efficient <i>A. tumefaciens</i> mediated genetic transformation ","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141516440","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":"Somatic embryogenesis induction in Rosa persica Michx ex Juss.","authors":"Shahrzad Vaziee, Mostafa K. Sarmast, Farshid Ghaderi-far, Changquang Wang","doi":"10.1007/s11240-024-02805-6","DOIUrl":"https://doi.org/10.1007/s11240-024-02805-6","url":null,"abstract":"<p>The current research aimed to evaluate the impact of various explants (immature zygote embryos, hypocotyls, and cotyledons), the effects of liquid and agar solidified MS media, as well as 2,4-D and BA on the induction and development of somatic embryos from <i>Rosa persica</i>. Although hypocotyls and cotyledon explants generated callus and primary globular embryos, only immature zygotic explants were able to produce mature somatic embryos in a half-strength liquid and agar- solidified MS medium. Indirect mature somatic embryos were only produced in agar-solidified MS media while liquid MS media-derived direct and indirect embryos could not further develop into mature embryo. Embryogenic callus induction mostly occcurred in half-strength MS media supplemented with 0.3 mg L^− 1 2,4-D or 0.09 mg L^− 1 BA and 0.3 mg l^− 1 2,4-D on half-strength MS liquid and agar-solidified media, respectively. Immature zygote embryos formed embryogenic calli at a success rate of 57.14% and 46% on half-strength MS agar-solidified and liquid media, respectively. Indirect Somatic embryogenesis was induced only from detached immature zygote explants, achieving a frequency of 25.11% and 9.64% in agar-solidified and liquid media, respectively. However, the concentration of PGR to induce the highest somatic embryo was different in liquid and agar-solidefied media. In liquid media, the incubation of explant for four weeks on half-strength MS media supplemented with 1 mg L^− 1 of BA and 0.9 mg L^− 1 2,4-D and then subculturing them in half-strength MS media supplemented with 1 mg L^− 1 of BA and 0.3 mg L^− 1 2,4-D resulted in the highest indirect somatic embryo (24.5 embryos per explant). The same MS media supplemented with 1 mg L^− 1 of BA and 0.09 mg L^− 1 2,4-D resulted in direct somatic embryo (4.3 embryos per explant). However, non of the somatic embryos could further germinate in liquid media. In agar-solidified media, the incubation of explants in half-strength MS media supplemented with 0.3 mg L^− 1 2,4-D during the induction phase and subsequent subculture of embryogenic callus on half-strength MS media supplemented with 1 mg L^− 1 of BA and 0.3 mg L^− 1 2,4-D resulted in the highest somatic embryo (26 somatic embryos per explant). The highest germination rate (46.15%) of somatic embryos occurred in PGR-free and agar-solidified MS media from 16-weeks-old embryogenic callus culture, which was primarily incubated in half-strength MS media supplemented with 0.3 mg L^− 1 2,4-D for four weeks and then subculturing them in half-strength MS media supplemented with 1 mg L^− 1 of BA and 0.3 mg L^− 1 2,4-D. The acclimatization efficiency of the acquired plantlets was 25%, and they could not grow vigorously in the greenhouse. The results of this experiment can help utilize wild relative <i>Rosa</i> species in breeding program","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141516375","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":"Qualitative and quantitative analysis of high-value metabolites from callus culture of Alcea rosea L.","authors":"Hasnat Tariq, Anisa Andleeb, Sannia Batool, Urooj Ali, Muhammad Waqas, Abdul Wahab, Gouhar Zaman, Samantha Drouet, Nathalie Giglioli-Guivarc’h, Christophe Hano, Bilal Haider Abbasi","doi":"10.1007/s11240-024-02797-3","DOIUrl":"https://doi.org/10.1007/s11240-024-02797-3","url":null,"abstract":"<p>This study explored how plant growth regulators (PGRs) impact callus induction, biomass accumulation, phytochemical synthesis, and the antioxidant and anti-aging potential of <i>Alcea rosea L.</i> callus cultures. Our findings revealed that the combination of 1 µM NAA and 0.1 µM TDZ leads to 100% callus induction frequency and the highest biomass accumulation, with a fresh weight of 267 g/L and a dry weight of 16.5 g/L. Maximum total phenolic (719.19 mg GAE/L) and total flavonoid (166.34 mg QE/L) production were observed in callus cultures treated with 1 µM NAA. High-performance liquid chromatography identified 9 anthocyanin compounds, with the highest total anthocyanin production (265.65 µM DW) recorded with a PGR concentration of 1 µM NAA + 0.1 µM TDZ. Antioxidant activities varied across different PGR concentrations, with notable values including DPPH-FRSA of 83.1%, ORAC value of 1089.13 µM TEAC, and maximum reduction in ROS/RNS levels by 87.56%. Anti-aging activities also varied, with tyrosinase inhibition peaking at 79.12%, vesperlysine-like AGEs’ inhibition at 68.32%, and pentosidine-like AGEs’ inhibition at 56.42%. This study introduces a novel approach that addresses traditional extraction method limitations by examining the impact of PGRs on callus culture initiation and secondary metabolite production, alongside antioxidative and anti-aging characteristics. Such methodologies potentially add to our understanding of <i>A. rosea L.</i>‘s therapeutic potential and contribute to sustainable techniques for producing bioactive compounds.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141516376","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":"Induction and characterization of polyploids through morpho-anatomical, cytological, chemotypic, and molecular approaches in Patchouli (Pogostemon cablin Benth.)","authors":"Channayya Hiremath, K. N. Prabhu, R. Ravi Kumar, T. K. Pranav Raj, Ram Swaroop Verma, Dinesh A. Nagegowda","doi":"10.1007/s11240-024-02798-2","DOIUrl":"https://doi.org/10.1007/s11240-024-02798-2","url":null,"abstract":"<p>Patchouli (<i>Pogostemon cablin</i>), is an industrially important aromatic plant that produces patchouli oil. The present experiment aimed to generate polyploid plants of patchouli with increased herb yield, oil production, and higher patchouli alcohol content. In the present study, colchicine, a chemical compound known to induce polyploidy was used at concentrations of 0.2% and 0.3% for 48 h. The diploid variety CIM-Shrestha (2n = 2X = 32) was used as the starting point. Polyploidy was successfully induced in vitro conditions and confirmed through various analyses including chromosome counting, anatomical, morphological, and gene expression studies. Treating the diploid callus with 0.2% colchicine resulted in viable and stable tetraploid seedlings. These tetraploids had a chromosomal count of 2n = 4x = 64. Tetraploids exhibited distinct morphological, anatomical, and chemical characteristics, such as thicker leaves, more oil glands, higher chloroplast numbers, larger stomata size, patchouli alcohol, and α-Guaiene. In field conditions, the induced tetraploids remained stable and the majority of tetraploid lines showed higher oil content and patchouli alcohol concentrations compared to diploid plants, although herb yield was lower in tetraploids. In the gene expression study, five (<i>PcHMGR</i>, <i>PcFPPS</i>, <i>PcTPSCF2</i>, <i>PcTPSB15</i>, and <i>PcPTS</i>, along with the expression of the control gene <i>Pc18S</i>) pathway genes related to the patchouli alcohol production pathway were studied. The Real Time-qPCR results show that there was no significant change in the expression of any of the five analyzed genes.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141255109","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":"Somatic embryogenesis of slash pine (Pinus elliottii Engelm.): initiation, maturation, germination and mycorrhization of regenerated plantlets","authors":"Qi Fei, You-Mei Chen, Xin Ke, Jian-Ren Ye, Li-Hua Zhu","doi":"10.1007/s11240-024-02789-3","DOIUrl":"https://doi.org/10.1007/s11240-024-02789-3","url":null,"abstract":"<p>Slash pine (<i>Pinus elliottii</i> Engelm.), an important economic tree species with a high resin yield, is extensively cultivated in southern China. Somatic embryogenesis (SE) technology could hasten the breeding of <i>P. elliottii</i> through improving seedling quality and number, but mass production of somatic embryos remains constrained by various factors. We described an efficient SE system through indirect route and investigate the effects of genotype, phytohormones and culture condition on SE. Immature zygotic embryos of three open-pollinated mother trees collected during 2020–2023 were utilized for initiation of pro-embryogenic masses (PEMs). The initiation of PEMs was significantly influenced by seed sources (families) (<i>p</i> &lt; 0.05). The addition of 2,4-dichlorophenoxyacetic acid (6 mg/L), 6-benzylaminopurine (1 mg/L) and kinetin (2 mg/L) effectively improved initiation rates by up to 15.67 ± 1.89%. Furthermore, glutamine addition to the maturation medium enhanced the yield of somatic embryos during culture. The optimum concentration range of activated carbon within the germination medium spanned from 3–4.5 g/L. Inoculation with ectomycorrhizal fungus <i>Laccaria amethystea</i> enhanced the root length of regenerated plantlets, reaching 7.92 ± 4.06 cm. The regenerated plantlets inoculated with <i>Pisolithus orientalis</i> and <i>Xerocomus chrysenteron</i> showed improved survival during the acclimatization phase, with 100% survival of mycorrhizae-treated plantlets after 3 months of acclimatization in the greenhouse. We clarified the SE conditions suitable for <i>P. elliottii</i>, and the SE system and mycorrhizal method we used can be applied to mass production of plantlets.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141255112","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":"Exogenous application of salicylic acid and low-dose ionizing radiation increases synthesis of bioactive compounds and upregulates phytosterol production of Bitter melon (Momordica charantia)","authors":"Saptaparna Dutta, Sankalan Dey, Anindita Chakraborty, Sarmistha Sen Raychaudhuri","doi":"10.1007/s11240-024-02792-8","DOIUrl":"https://doi.org/10.1007/s11240-024-02792-8","url":null,"abstract":"<p>Gamma radiation (GR) and salicylic acid (SA) enhance plant traits by regulating growth and development. However, the underlying mechanism of their combined therapies in plants is not well understood. In our study, the potential impact of 100 μM and 500 μM SA and/or 20, 40 Gy in <i>Momordica charantia</i>, a medicinal crop plant was assessed. We observed significant increase in plant root-shoot length and photosynthetic efficiency at 20 Gy dose, which further enhanced under SA application. Overall, simultaneous application of SA and GR also upregulated antioxidant capacity via production of secondary metabolites, such as flavonoids and phenolics. The highest antidiabetic activity with up to 81.5% inhibition of alpha-amylase enzyme was observed at 20 Gy + 500 μM SA combination. The expression of <i>Sterol C-22 Desaturase,</i> a key gene involved in the biosynthesis of β-sitosterol was upregulated under the treatment of GR and/or SA. β-sitosterol is an important compound with potent anti-cholesterol and anti-carcinogenic properties. Our study showed that highest content of β-sitosterol (1082.2 ± 68.2 ng/mg FW) was observed in <i>M. charantia</i> plants exposed to combined 20 Gy + 500 μM SA treatment. Therefore, gamma irradiation and salicylic acid can help in sustainable growth and production of various secondary metabolites in <i>M. charantia</i>, thereby enhancing its agronomic value.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141170928","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":"6-benzylaminopurine induces somatic embryogenesis in the staminodia of new genotypes of Theobroma cacao L. from the Papaloapan Basin of Mexico and differs from that of T. bicolor Bonpl.","authors":"Nancy Carmín Martínez-Hernández, Carlos Antonio Dávila-Figueroa, José Antonio Morales-Serna, Karla Viridiana Castro-Cerritos, Nelda Xanath Martínez-Galero, Héctor López-Arjona, Enrique Villalobos-Amador","doi":"10.1007/s11240-024-02783-9","DOIUrl":"https://doi.org/10.1007/s11240-024-02783-9","url":null,"abstract":"<p><i>Theobroma cacao</i> and <i>T. bicolor</i> are among the most important agricultural crops of the Mexican tropics. Currently, in Mexico, the propagation of these crops is performed via seeds, which indicates that demand exceeds production. In this context, somatic embryogenesis (SE) is an alternative to this approach. Thus, we evaluated the presence of embryogenic genotypes of <i>T. cacao</i> and <i>T. bicolor</i> in the Papaloapan Basin of Mexico with the idea of implementing this technology in the region. The analysis of the phenotypic expression of the floral whorls demonstrated that, unlike <i>T. bicolor</i>, the combination of 6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid induces different morphogenetic responses in the genotypes that were evaluated in the primary callus phase of <i>T. cacao</i>. Staminodia presented with the highest percentage of caulogenesis in <i>T. cacao</i>, whereas <i>T. bicolor</i> presented with the highest frequency of caulogenesis in Staminodia and carpels. Some calli differentiated in the roots to a greater extent than those derived from the staminodes of <i>T. cacao</i>. The calli of <i>T. bicolor</i> did not differentiate. A parallel study using thidiazuron as an inducer demonstrated similar results for calli of both species; however, rhizogenesis from staminodes was 50% lower for the evaluated genotypes of <i>T. cacao</i>. Staminodes were the only structures that demonstrated primary somatic embryogenesis (PSE) in 66% of the <i>T. cacao</i> genotypes that were evaluated by using benzylaminopurine. Finally, secondary somatic embryogenesis (SSE) was evaluated in cotyledons and reached a 60% success rate, of which 95.48% were normal somatic embryos. Both types of embryogenesis were morphologically characterized using optical and/or scanning electron microscopy.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141170944","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":"Using hyperspectral signatures for predicting foliar nitrogen and calcium content of tissue cultured little-leaf mockorange (Philadelphus microphyllus A. Gray) shoots","authors":"Razieh Khajehyar, Milad Vahidi, Robert Tripepi","doi":"10.1007/s11240-024-02765-x","DOIUrl":"https://doi.org/10.1007/s11240-024-02765-x","url":null,"abstract":"<p>Determining foliar mineral status of tissue cultured shoots can be costly and time consuming, yet hyperspectral signatures might be useful for determining mineral contents of these shoots. In this study, hyperspectral signatures were acquired from tissue cultured little-leaf mockorange (<i>Philadelphus microphillus</i>) shoots to determine the feasibility of using this technology to predict foliar nitrogen and calcium contents. After using a spectroradiometer to take hyperspectral images for determining foliar N and Ca contents, the correlation between the hyperspectral bands, vegetation indices, and hyperspectral features were calculated from the spectra. Features with high correlations were selected to develop the models via different regression methods including linear, random forest (RF), and support vector machines. The results showed that non-linear regression models developed through machine learning techniques, including RF methods and support vector machines provided satisfactory prediction models with high R² values (%N by RF with R² = 0.72, and %Ca by RF with R² = 0.99), that can estimate nitrogen and calcium content of little-leaf mockorange shoots grown in vitro. Overall, the RF regression method provided the most accurate and satisfactory models for both foliar N and Ca estimation of little-leaf mockorange shoots grown in tissue culture.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141171042","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":"Hairy root culture of Strobilanthes cusia for the production and enhancement of indigo biosynthesis","authors":"Rajkumari Lunphasana Devi, Sanasam Thoibi Devi, Keithellakpam Sanatombi","doi":"10.1007/s11240-024-02791-9","DOIUrl":"https://doi.org/10.1007/s11240-024-02791-9","url":null,"abstract":"<p><i>Strobilanthes cusia</i> is an indigo-yielding plant and scarcity of both wild and cultivated plants has increased the supply pressure of the natural dye. Since hairy root cultures are suitable for the in vitro production of secondary metabolites, the hairy root culture of <i>Strobilanthes cusia</i> was established using two <i>Rhizobium rhizogenes</i> strains (ATCC 15834 and MTCC 532). Polymerase chain reaction (PCR) using <i>rol B</i> primer confirmed the molecular evidence of hairy root transformation. The differential transformation frequency was studied based on factors like explants types (leaves, shoot tips, and stem); infection techniques, infection period, co-cultivation period, bacterial strain types, acetosyringone concentration, and antibiotic treatment conditions. Leaf explants, infected with ATCC 15834 exhibited the highest induction rate (58.67%) among the three explants. The most efficient hairy root transformation was achieved with the immersion technique, involving 10 min of infection and 5 days of co-cultivation using 100 µM acetosyringone during infection. When the indigo content of the hairy roots and different parts of in vitro plantlets were compared, the hairy roots produced 0.314 mg/g fresh weight (FW) of indigo, slightly more than the roots (0.21 mg/g FW), but less than the leaves (1.08 mg/g FW) of in vitro plantlets. Further, the indigo content of the hairy root cultures treated with 200 µM indican and 200 µM salicylic acid were enhanced up to 0.78 mg/g FW and 0.71 mg/g FW, respectively. Thus, this study reveals the potential of hairy roots of <i>S. cusia</i> for indigo biosynthesis, which may serve as a potential alternative source for natural indigo.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141516325","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 and application of the TRV-induced gene-silencing system in different Rhododendron species","authors":"Yunyun Xu, Yumeng Cui, Hanyu Chen, Ya Pu, Chunying Zhang, He Huang","doi":"10.1007/s11240-024-02782-w","DOIUrl":"https://doi.org/10.1007/s11240-024-02782-w","url":null,"abstract":"<p><i>Rhododendron</i> is one of the top ten traditional flowers in China, with high ornamental and medicinal values. However, molecular research on this species has been impeded by the lack of efficient molecular genetic techniques. Virus-induced gene silencing (VIGS) technology is an effective tool for analyzing gene functions, which has been successfully applied to many plant species. But there have been no reports of VIGS system for <i>Rhododendron</i>. In this study, tobacco rattle virus (TRV) was chosen to infect <i>Rhododendron micranthum</i> using <i>phytoene desaturase</i> (<i>PDS</i>) as the reporter gene. After the inoculation of pTRV2-<i>RmPDS</i> by leaf injection, photobleaching phenomena appeared in the newly developed leaves and the RT-qPCR assay demonstrated that <i>RmPDS</i> was successfully silenced. Then three parameters including the developmental stage, the <i>Agrobacterium</i> concentration and the inoculation temperature were examined to optimize the system. The silencing efficiency was increased from 2.4 to 11.4% and the optimized conditions were as follows: the developmental stage of the two true-leaf stage, the adjustment of the inoculation solution to a final OD₆₀₀ = 1.5 and the inoculation temperature of 18 ℃. To further validate the system, the most optimal combination was used to infect other six rhododendron genotypes. <i>R. mucronulatum</i>, <i>R. ovatum</i>, <i>R.</i> × <i>pulchrum</i>, <i>R. simsii</i> and <i>R. yedoense</i> displayed the silenced phenotype of <i>PDS</i> as expected. We successfully established TRV-mediated VIGS technology in <i>Rhododendron</i> which could evaluate and characterize the function of plant genes without the need for cumbersome tissue culture.</p>","PeriodicalId":20219,"journal":{"name":"Plant Cell, Tissue and Organ Culture","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141170841","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}