Plant Cell, Tissue and Organ Culture最新文献

Nanomaterials usually have specific characteristics due to their incredibly tiny size, which also increases their surface area, providing a more interactive surface. Compared to their macro-sized counterparts, these tiny nanoparticles exhibit a multitude of size-dependent properties. Plant tissue culture (PTC) plays an important role in bioactive chemical synthesis, mass cultivation, protection, genetic control, and plant enhancement. Different nanoparticles (NPs) are utilized to improve the tissue culture responses of explants. Various nanoparticles, including cobalt, copper, silver, gold, zinc, selenium, titanium, iron, palladium, cerium, indium, manganese, aluminum, barium, silicon, nickel, zirconium, and their oxides, are used in this regard. Nowadays, it is critical to use nanosystems in conjunction with PTC for mass reproduction, conservation, genetic engineering, crop enhancement, and the synthesis of bioactive compounds. Nanostructured metal oxides play an important role in in vitro plant cultivation. The use of metal nanoparticles (MNPs) has successfully removed microbial contaminants from explants and had a favorable impact on organogenesis (increasing the growth of shoots, roots, and multiplication ratios), callus induction, metabolic changes, and the synthesis of secondary metabolites (NPs are used as elicitors or stress agents). Additionally, NPs cause somaclonal variation (modifications to DNA), improve cryopreservation (increasing the survival rate), and enhance genetic transformation (facilitating gene transformation to bypass the plant cell wall barrier and accelerating protoplast isolation). This review aims to summarize the current breakthroughs achieved by integrating nanotechnology with PTC.

Graphical abstract

Neolamarckia cadamba (N. cadamba) is an evergreen tree species known for its rapid growth, remarkable wood properties, and significant value in medicine, feeding, and landscape. In order to clone a N. cadamba individual with excellent genotype, a plant regeneration protocol was successfully established with leaves wrapped by bud scales as explants. The optimal sterilization method for the leaves was 0.1% Mercury Chloride (HgCl₂) treatment for 1 min before culturing on Murashige and Skoog’s medium (MS) supplemented with 3.0 mg/L Thidiazuron (TDZ), 0.1 mg/L 2–4 Dichlorophenoxyacetic acid (2-4D), 0.05 mg/L α-Naphthaleneacetic acid (NAA) and 1 mL/L Plant Preservative Mixture (PPM) to induce calluses. The medium containing 1 mL/L PPM could effectively inhibit explant contamination without an unfavorable impact on the final induction rate of callus from the leaves. Three types of calluses were induced from the leaves cultured on the above medium. Among them, only the Type II callus, which was green and nodular, had few particle masses, could differentiate into adventitious shoots on the MS medium supplemented with 1.5 mg/L 6–Benzylaminopurine (6-BA) and 0.05 mg/L NAA, with the induction rate of 78.89% and adventitious shoot number per callus of 11.67. The adventitious shoots were proliferated on the MS medium supplemented with 1.0 mg/L 6-BA and 0.05 mg/L Indole-3- butyric acid (IBA) with the proliferation coefficient of 3.37. And the micro-shoots developed roots in the MS medium supplemented with 0.05 mg/L NAA and 0.05 mg/L IBA. The regeneration protocol can be used in the propagation and large scale production of seedlings with the same genotype as an excellent individual of N. cadamba in the field.

Polyploidy induction is a promising strategy for enhancing stress tolerance and physiological resilience in crop plants. In this study, we investigated the efficacy of oryzalin treatment in inducing autoploidy in the finger lime hybrid 'UF SunLime' and examined its implication for cold tolerance and physiological responses to stress. Murashige and Skoog (MS) medium supplemented with oryzalin (0, 4, 6, and 8 mg/L) was applied to germinated seeds or adventitious shoots for 3 or 6 days, and the optimal conditions for maximal tetraploid induction were determined. Ploidy analysis using flow cytometry confirmed the ploidy level of the generated seedlings and shoots. Oryzalin applied at 6 mg/L for 6 days was the most effective concentration at generating the highest percentage of autotetraploids (60%), followed by 6 mg/L for 3 days (40%). Phenotypic characterization revealed significant differences in leaf morphology and chlorophyll content between tetraploid and diploid plants. Moreover, tetraploid plants exhibited decreased water loss and electrolyte leakage during cold stress assessments compared to their diploid counterparts. Pigment content analysis indicated differential responses to cold stress, with tetraploid plants generally exhibiting greater total phenolic compound content. Differential expression of transcription factors related to cold stress were identified as contributing factors to the cold tolerance induced by tetraploidy. Overall, our findings highlight the successful induction of autoploidy in the 'UF SunLime' finger lime hybrid cultivar via oryzalin treatment and provide insights into the physiological responses and stress tolerance conferred by polyploidization in citrus plants.

The spectral quality of light regulates plant growth through a variety of physiological, biochemical, and molecular processes. Earlier research on ginseng sprouts have largely focused on the impact of monochromatic lights, with limited attention to combinations with white light. The present study explored the influence of white light supplemented with different wavelengths on ginseng sprout growth at both low (30) and high (200) µmol m^− 2 s^− 1 PPFD intensities. Significant variations in growth were noted between the two light intensities. Specifically, the green light combination proved advantageous for enhancing both growth and photosynthesis across both intensity levels. The chlorophyll fluorescence characteristics, stomatal properties, total soluble sugars, total soluble proteins, and root activity were notably enhanced by W + G (200 PPFD) treatment. Enzymatic and non-enzymatic antioxidant activities were predominantly influenced by W + R treatments at both intensity levels. Low light, particularly, W + R (30 PPFD) and W + G (30 PPFD) significantly increased mineral content in both the shoot and root. Low light intensities positively influenced the accumulation of ginsenosides Rd, F₂, and Rg₁ in treatments W, W + R, and W + G, respectively while Rd₂ accumulation was greatly promoted by high intensity white light treatment, W (200 PPFD). Gene expression involved in the ginsenoside biosynthesis pathway aligned with ginsenoside accumulation. Therefore, to meet the growing demand for ginseng sprouts, optimizing light quality, particularly by incorporating green or red light in conjunction with white light, might serve as a promising approach for enhancing both the quality and yield of ginseng sprouts within the controlled environment agriculture system.

Withania somnifera (L.) Dunal, an Ayurvedic herb rich in withanosides, has anti-cancer and health-rejuvenating properties. These bioactive chemicals, primarily found in the roots, are of significant interest for production in hairy root cultures. In this study, researchers investigated a hairy root line produced by Rhizobium rhizogenes (ATCC 43,057), varying cell density, infection period, and acetosyringone concentrations. The most successful root induction frequency (31.66% ± 2.98%) occurred at a bacterial cell density of OD₆₀₀ = 0.8 combined with a 20-min infection period and 200 µM acetosyringone. After 25 days, the hairy root culture achieved its highest biomass (5.20 ± 0.15 g/50 mL flask fresh weight (FW)) in MS liquid medium with 3% sucrose. At 30 and 25 days, the greatest amounts of withanolide A (0.824 ± 0.01 mg/g dry weight (DW) and withaferin A (1.292 ± 0.02 mg/g DW) were observed. Hairy roots were further treated with methyl jasmonate (MJ) and salicylic acid (SA) at various concentrations (25, 50, and 100 µM). Notably, the maximum content of withanolide A (2.90 ± 0.08 mg/g DW, 3.86-fold) and withaferin A (3.40 ± 0.01 mg/g DW, 2.61-fold) were obtained in the hairy roots treated with MJ at 100 µM. After 48 h, the content of withanolide A reached 3.16 ± 0.03 mg/g DW (6.72-fold), and withaferin A peaked at 4.32 ± 0.05 mg/g DW (4.59-fold) when treated with 50 µM SA + 100 µM MJ. This study demonstrated that W. somnifera hairy root culture enhances large-scale production of withanolide A and withaferin A.

Betalains are nitrogen-containing natural pigments that are water soluble and they comprise of the red-violet betacyanin and the yellow betaxanthin which are abundant in plants such as red/yellow beet, amaranth, prickly pear, pitaya, and others. They are widely used as food coloring agents for many centuries. Betalains are used in pharmaceuticals, functional foods, and cosmeceuticals, since they have tremendous potential to scavenge free radicals and prevent diseases, such as hypertension, dyslipidemia, cancer, neurological disorders, and vascular stenosis. Betalains are proven to be toxicologically safe and have health benefits, they have been approved as food additives in the United States of America, and European countries. Although betalains can be found in natural resources, there are differences in their composition, amounts, and seasonality. For this reason, researchers have developed alternative methods of producing these valuable compounds using cell and organ culture techniques. In several plants, cell and organ cultures are established, and bioreactor technologies have been used to produce betalains on a wide scale. In this review, we discuss the varied biotechnological methods and approaches applied for the biosynthesis of betalains including metabolic engineering approaches.

Prosthechea cochleata or clamshell orchid is recognized as a species of both conservation and commercial importance. It has long been prized by orchid breeders and growers for its unique flower and hardy disposition. The commercial market has failed to meet the demand for this species since it is still targeted for illegal collection in the wild. This study examines the effectiveness of the banana powder on two variants (var. diandra and var. triandra) with disparate home ranges and levels of genetic diversity using two commercially available agar-based media (PhytoTechnology P668 and P748). Undifferentiated protocorms of the two variants were monitored for shoot growth and differentiation across a period of 156 days. The growth media with 3% banana powder supplement (P748) was more effective in initiating shoot formation in both P. cochleata variants compared to control media (P668) (F = 27.15, p < 0.001, df = 81). The diandrous variant grown with banana showed the highest mean shoot count (28.67 shoots/flask) at the end of the monitoring period. Results suggest that banana supplement is an effective source of plant growth regulators and organic nutrients necessary to promote shoot formation and seedling development in this species. This research is important for the conservation of P. cochleata as it identifies an effective and cost-efficient method for in vitro propagation, which can aid in increasing the ex-situ and in situ populations of this threatened species. Additionally, the findings have implications for commercial orchid production, as it can potentially improve the cultivation and yield of this species for economic purposes.

This work presents an efficient one-step procedure for in vitro propagation in Angelica glauca using rhizome buds and production of secondary metabolites. A maximum of 94% of buds were established in vitro on medium supplemented with 0.3 mg/L Benzyl adenine (BA) and 0.1 mg/L Gibberellic acid (GA₃). After the fifth sub-culture, the proliferating shoots from the rhizome buds displayed the maximum proliferation (1:15), rooted on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BA and 0.2 mg/L α-naphthalene acetic acid (NAA). After being transferred to pots with soil:cocopeat (1:1) for hardening, shoots with enlarged rhizomes demonstrated 60% survival after a month in the polyhouse. For secondary metabolite production, callus was induced from in vivo roots on MS medium supplemented with 0.5 mg/L Kinetin (Kin) and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) under dark incubation and after 1 year its suspension culture showed the existence of 206 compounds. The gas chromatography-mass spectrometry (GC–MS) analysis results revealed that extracts predominantly contain compounds from different classes such as esters, ethers, and N-heterocyclic pyrrolo pyridazine, fatty acids and mono and sesquiterpenes with varying concentrations. On elicitation with 0.5, 1.0 and 1.5 mM Methyl jasmonate (MeJA) the callus cultures depicted varying concentration of monoterpene such as d-limonene, trans and cis-ligustilide, a marker compound of A. glauca essential oil, fatty acids and ethers. Sucrose treatment at 1, 3 and 5% revealed the presence of various unsaturated fatty acids, hydrocarbon, ethers, sesquiterpenes β-farnesene, α-copaene, and carotenoid rhodopin. Addition of growth regulators (2,4-D and Kin) revealed the presence of furfural and its derivatives, benzoic acids and esters.

Puya chilensis is an endemic species of Chile belonging to the Bromeliaceae family, known for its morphological plasticity and ecological importance. In recent years, greater attention has been given to its conservation due to the indiscriminate use of its populations for food and medicinal purposes. In vitro culture is a tool for the propagation of plants species, as well as for obtaining bioactive compounds. In Chile, P. chilensis is the most representative within the Puya genus. This study focused on establishing an in vitro propagation protocol for P. chilensis that allows massive propagation and obtaining proteolytic enzymes. The combined effect of plants cuts and the application of 0.5 µmol L⁻¹ of BAP and GA₃ favored the in vitro multiplication of P. chilensis. Culture in liquid medium induced greater morphological development and early differentiation of anatomical structures in the leaves of P. chilensis in vitro. The use of TIS creates ideal conditions during the last stage of in vitro culture and ensures 100% survival during acclimatization phase. The management of cultivation conditions and the efficient use of TIS allowed the generation of P. chilensis plants with an optimal degree of development for obtaining proteolytic extracts. The main enzymes present in the extracts of P. chilensis plants grown in vitro belong to the cysteine type. This study proposes for the first time an optimized protocol for the propagation and conservation of P. chilensis, enhancing its uses as a source of biologically active molecules for the biotechnology and pharmaceutical industries.

Graphical Abstract

Cryopreservation, a widely utilized technique for the long-term preservation of in vitro cultures, effectively arrests metabolic processes, obviating the need for frequent subcultures and mitigating the risk of somaclonal variation. In this study, we applied cryopreservation methods to intact rice (Oryza sativa L.) calli to determine the optimal age for cryopreservation, investigating the timelines for greening and shoot initiation in R0 plants. Results revealed that three-month-old calli exhibited the highest regeneration percentage, with greening observed within twelve days and shoot initiation within fifteen days. Using 3% mannitol in the callus culture medium provided osmotic stress, aiding in the formation of compact calli masses suitable for regeneration. Vitrification with cryoprotectants (DMSO, PEG, and glucose) and gradual dehydration improved cell survival. Thawing and post-thaw damage were minimized using rapid thawing, fast cryoprotectant removal, and gradual rehydration. We assessed the phenotypic variations in R1 and R2 generation and genotypic fidelity of regenerants in R1. Phenotypic variations from seed-derived plants were observed in seed characters both in vitrified and cryopreserved calli-derived plants. However, these variations were unstable and disappeared in the R2. SSR markers were used to detect genetic variations in R1, with results showing a 3.6% change in vitrified calli-derived plants and an 8.61% change in cryopreservation-derived plants, likely due to reversible DNA methylation or SNPs in non-coding region. Our study confirms the feasibility of cryopreservation for rice calli, ensuring high regeneration rates and minimal long-term genetic variations.