Synthetic Seeds Research Articles

Standardization of Protocol for Production of Synthetic Seed of Brahmi (Bacopa monnieri) and Assessment of their Longevity under Differential Storage Condition

Brahmi [Bacopa monnieri (L) Pennel] is an important medicinal small succulent, medicinal herb belongs to the family ‘Scrophulariaceae’ which is commonly known as Jala Brahmi or Nira-Brahmi. National Medicinal Plants Board (NMPB), estimated an annual demand of Bacopa in 2004-2005 was 6621.8 tonnes, with a 7% annual growth rate. Micropropagation techniques and production of synthetic seeds can be a breakthrough to overcome the low availability of Brahmi propagating materials. So, the present study was conducted to standardize the protocol for preparation of synthetic seeds of Brahmi using nodal segment. The experiment was conducted with five treatments by using combinations of different concentrations of sodium alginate and calcium chloride and preparing two types of synthetic seeds (single layer and double layer). Further the longevity of both single layer and double layered synthetic seeds was studied under two different storage temperatures at 25°C (ambient condition) and 4°C (refrigerator) up to 40 days by inoculating them in two different types of media. The result showed no significant effect of concentrations of gelling agent and complexing agent on the germination percentage of synthetic seeds. The number of shoots per synthetic seed varied from 22.00 to 29.35 with 100% germination and maximum shoots/synthetic seed and shoot length was obtained by using T5 (single layered with MS media, 3% sodium alginate and 2.5% calcium chloride). Maximum number of days of storage can be observed in the single layer synthetic seeds when stored at 4°C along with media followed by double layer synthetic seeds when stored at 4°C along with media. However, seeds stored on vermiculite got dried up in the initial days of storage. Germination was found to be higher in seeds stored at 4°C when compared to seeds stored at 25°C. The study as a whole elucidated the opportunity for the use of synthetic seed as an alternative to traditional propagation system for Brahmi.

TC‐GEN: Data‐Driven Tropical Cyclone Downscaling Using Machine Learning‐Based High‐Resolution Weather Model

AbstractSynthetic downscaling of tropical cyclones (TCs) is critically important to estimate the long‐term hazard of rare high‐impact storm events. Existing downscaling approaches rely on statistical or statistical‐deterministic models that are capable of generating large samples of synthetic storms with characteristics similar to observed storms. However, these models do not capture the complex two‐way interactions between a storm and its environment. In addition, these approaches either necessitate a separate TC size model to simulate storm size or involve post‐processing to capture the asymmetries in the simulated surface wind. In this study, we present an innovative data‐driven approach for TC synthetic downscaling. Using a machine learning‐based high‐resolution global weather model (ML‐GWM), our approach can simulate the full life cycle of a storm with asymmetric surface wind that accounts for the two‐way interactions between the storm and its environment. This approach consists of multiple components: a data‐driven model for generating synthetic TC seeds, a blending method that seamlessly integrates storm seeds into the surrounding while maintaining the seed structure, and a model based on a recurrent neural network to correct for biases in storm intensity. Compared to observations and synthetic storms simulated using existing statistical‐deterministic and statistical downscaling approaches, our method shows the ability to effectively capture many aspects of TC statistics, including track density, landfall frequency, landfall intensity, and outermost wind extent. Leveraging the computational efficiency of ML‐GWM, our approach shows substantial potential for TC regional hazard and risk assessment.

Blackberry synthetic seeds storage: effects of temperature, time, and sowing substrate

In vitro propagation, is becoming the predominant method for blackberry propagation due to its advantages compared to agamic traditional propagation methods. Synthetic seed technology represents a promising approach to further enhance the productivity of in vitro propagation facilitating the exchange of plant materials among laboratories and contributing to germplasm conservation efforts. This study aimed to establish an optimal protocol for the storage and sowing of synthetic blackberry seeds obtained through the encapsulation of clump bases. The synthetic seeds were sown without storage (Control) and after storage periods of 30, 60, and 120 days at 4 °C and 25 °C in the dark, in three different substrates (agarised, perlite, and potting). After forty-five days from sowing viability, regeneration rate, shoot and root numbers and lengths, as well as fresh and dry weights of the plantlets, were assessed. Results indicated that agarised substrate consistently exhibited favourable outcomes, with sustained regeneration rates and robust plantlet development even after prolonged storage at 4 °C. Synthetic seeds sown in perlite and potting substrates demonstrated enhanced regeneration rates following storage at 4 °C for 60 and 120 days. On the contrary, storage at 25 °C resulted in a notable decline in regeneration rate, highlighting its inadequacy for blackberry synthetic seed conservation purposes. These findings underscore the importance of sowing substrate selection and storage temperature in optimizing the storage and sowing protocols for synthetic blackberry seeds.

A research on the production, storage and germination of synthetic seeds in tea plant (Camellia sinensis [L.] O. Kuntze)

Tea plant, is one of the most popular beverages consumed worldwide because of its rich and pleasant flavors and numerous health benefits. In this study, we performed production, storage and germination of synthetic seeds in the tea plant by encapsulation of somatic embryos. In our research, after the encapsulation of the mass-produced embryogenic calli with different doses of sodium alginate (NaAlg) and CaCl2, they were stored at different temperatures and at different times, and then transferred to different nutrient media after the expiry of the different storage period to determine the most suitable nutrient composition for germination. The resulting embryogenic calli were stored after encapsulation and then transferred to germination media. Although there was very little germination in long-term storage, the data obtained were found to be statistically insignificant. Although there was very little germination in long-term storage, the data obtained were found to be statistically insignificant. The germination rate of the beads, which were transferred directly to the germination medium without storage and encapsulated using 3% Na-Alg and 50 mM CaCl2.2H2O, was determined as 44.44% in the MS medium containing 3 mg/L BAP and 1 mg/L IBA. In addition, it was observed in the study that increasing storage time increased the darkening of the beads, while increasing NaAlg and CaCl2 doses caused obtaining harder and more nontransparent beads. Hyperhydricity problem was not encountered in any trial in the study. This study, carried out with our local tea variety, is a pioneer in its field with the scope of encapsulating somatic embryos.

Synthetic Seed Production and Slow Growth Storage of In Vitro Cultured Plants of Iris pallida Lam.

Iris pallida Lam. is traditionally cultivated in Italy to sell its rhizomes to perfume-producing industries and is particularly sought-after because of its high content of irones, ketone compounds responsible for the violet smell of the orris essence. One of the critical aspects of its cultivation is the propagation method, performed by subdividing and replanting sections of the rhizome, which leads to the sacrifice of salable material. A solution is provided via in vitro propagation using the somatic embryogenesis technique, an effective method that allows the production of plants without the use of the rhizome. To facilitate the scale up of the activities of micropropagation companies, the method of slow growth storage (SGS) for orris plantlets and a somatic embryo encapsulation technique were developed for the first time. Orris plantlets were placed at 4 °C in the dark for 30, 60, 90 and 120 days and monitored 7 and 30 days after treatment. Synthetic seeds were obtained by encapsulating somatic orris embryos in sodium alginate beads, which were stored for 14 and 28 days at 4 °C and 24 °C. The results showed that it is possible to cold-preserve orris plantlets for up to 90 days without significant damages and that orris synthetic seeds can be produced and stored for a short-to-mid-term period. These conservation techniques can be useful for germplasm conservation and can also be integrated in the micropropagation cycle of orris, helping to solve issues related to the traditional propagation method.

Low-temperature storage in dark condition improved the in vitro regeneration of Plumbago zeylanica synthetic seeds: a medicinally valuable species

Low-temperature storage in dark condition improved the in vitro regeneration of Plumbago zeylanica synthetic seeds: a medicinally valuable species

Autotetraploid synthetic seeds of Ficus carica L. exhibit a superior cold acclimation system than their corresponding diploids

Autotetraploid synthetic seeds of Ficus carica L. exhibit a superior cold acclimation system than their corresponding diploids

The Influence of the Explant's Type on the Performance of Synthetic Seeds of Blackberry (Rubus spp.).

In vitro propagation, also known as micropropagation, has become the most widely employed method for blackberry propagation, as it overcomes the limitations of the traditional asexual propagation methods (mainly layering and cutting). In this context, synthetic seed technology represents a strategy to enhance the productivity of in vitro propagation and facilitates the exchange of plant materials between laboratories, contributing to germplasm conservation. This study aimed to identify the most suitable vegetative propagule for the encapsulation of blackberry. To this end, uninodal microcuttings (nodes) and the base of clumps were used to produce synthetic seeds for the cultivars Thornfree and Chester. Forty-five days after sowing, viability (percentage of green propagules without browning or necrosis), regeneration (percentage of propagules that sprouted and rooted simultaneously), number of shoots produced, shoot length, number of roots produced, root length, and the fresh and dry weights of the plantlets were measured. The results demonstrated that both considered propagules allowed us to obtain satisfactory regeneration rates. However, plantlets originating from the encapsulated clump's base had more shoots and roots, resulting in greater fresh and dry weights than the plantlets derived from encapsulated nodes. Therefore, for achieving more robust plantlets and enhancing overall procedural efficiency, we recommend using the base of clumps as a propagule for blackberry encapsulation.

Application of Encapsulation Technology: In Vitro Screening of Two Ficus carica L. Genotypes under Different NaCl Concentrations

Salinity stress represents an increasing issue for agriculture and has a great negative impact on plant growth and crop production. The selection of genotypes able to tolerate salt stress could be a suitable solution to overcome the problem. In this context, in vitro cultures can represent a tool for identifying the NaCl tolerant genotypes and quickly producing large populations of them. The possibility of exerting selection for tolerance to NaCl by using encapsulation technology was investigated in two genotypes of fig: ‘Houmairi’ and ‘Palazzo’. The effects of five concentrations of NaCl (0, 50, 100, 150 and 200 mM) added to the artificial endosperm were tested on the conversion of synthetic seeds and on the growth of derived shoots/plantlets. Moreover, proline (Pro) and malondialdehyde (MDA), the enzymatic activities of catalase (CAT), guaiacol peroxidase (POD), and EL (Electrolytic Leakage), as well as the chlorophyll content, flavanols, anthocyanins, and Nitrogen Balance Index (NBI) were determined on shoots/plantlet. The obtained results clearly showed that ‘Houmairi’ and ‘Palazzo’ could tolerate salt stress, although a strong difference was found depending on each specific physiological pathway. Indeed, ‘Houmairi’ was revealed to be more tolerant than ‘Palazzo’, with different response mechanisms to salt stress. The use of encapsulated vitro-derived explants proved to be a useful method to validate the selection of genotypes tolerant to salinity stress. Further investigation in the field must validate and confirm the legitimacy of the approach.

A new methodology to characterize the kinetics of a seed during the imbibition process

AimsAssuming the saturated, θs, and residual θr volumetric water contents of a seed as known inputs, we present a methodology to determine the hydraulic properties of a seed: α, n parameters and hydraulic conductivity Ks.MethodsThe seed is considered as a porous material in which water flow is governed with the same hydraulic properties defined for soils. Using the HYDRUS-2D software, the hydraulic properties of a seed were estimated from the inverse analysis of several cumulative seed imbibition curves measured at different seed water potentials, h. The optimum number of h was evaluated on synthetic seeds. The theoretical analysis was validated in laboratory experiments on barley, wheat and vetch seeds, where imbibition curves were measured with germination tests at seven levels of h (from 0 to -2.50 MPa).ResultsThe theoretical analysis showed that accurate estimates of α, n and Ks can be obtained if the most negative h-values are included in the optimization. The sensitivity analysis showed that the method allows obtaining a unique solution of α, n and Ks. The optimization error on the theoretical α, n and Ks was less than 1%. A satisfactory validation was also obtained on the experimental seed imbibition curves, with robust fits between the measured and optimized data. A unique solution of α, n and Ks was also obtained in all cases.ConclusionsA new method to determine the hydraulic properties of a seed is presented. This methodology could be used in different areas involving seed imbibition and also to simulate seed imbibition in different scenarios.

A liquid culture system for plantlet conversion and slow growth storage of encapsulated shoot tips of Justicia adhatoda L.

The present article demonstrates the applicability of liquid medium in propagation and short-term storage of synthetic seed of Justicia adhatoda L., a medicinally important plant used in many ayurvedic drugs. Shoot tips and nodal segments excised from shoots regenerated on meta-topolin (mT) containing medium were encapsulated in 3% Na-alginate and 100 mM CaCl2.2 H2O for the preparation of artificial seeds. Irrespective of media composition, shoot tips performed better than nodal segments with respect to shoot emergence from alginate beads and further conversion into plantlets in a single step. Morphogenic responses of encapsulated shoot tips in liquid as well as on agar-solidified medium were evaluated and maximum plantlet conversion was observed on PGR-free MS liquid medium. Encapsulated shoot tips were stored for 60 days using slow-growth storage approaches which were induced by a reduction of sucrose concentration (1%) in the medium and reducing MS medium strength to half. After storage, more than 50% synthetic seeds were converted into plantlets when transferred to a regrowth medium. This slow-growth approach may be applied for the short-term storage of synthetic seeds for a reasonable period that is sufficient for the exchange of germplasm for distant places.

In vitro regeneration, phytochemical profiling and antioxidant activity in Ruta chalepensis plants established from alginate encapsulated synthetic seeds

We developed a highly efficient protocol for the short-term conservation and in vitro regeneration of Ruta chalepensis L. This involved encapsulating nodal segments (NS) in a sodium-alginate solution (3% w/v) and calcium chloride (100 mM) to produce synthetic seeds. The effect of various concentrations of plant growth regulators (PGRs) on the conversion of synthetic seeds into plantlets was examined and the best results, with a conversion rate of 96.8%, were obtained on Murashige and Skoog (MS) medium supplemented with 5.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA) after 12 weeks of culture. When stored at 4 °C for different durations (2-12 weeks), encapsulated and non-encapsulated NS displayed distinct conversion responses. Encapsulated NS stored at 4 °C for 10 weeks exhibited a regeneration rate of 72.7%, while non-encapsulated NS did not survive under the same conditions. The proliferated shootlets derived from cold-stored NS were successfully rooted in agar-solidified MS medium containing 0.5 μM indole-3-butyric acid (IBA) and successfully acclimatized to ex vitro conditions. UV-Visible spectroscopy (UV), gas chromatography-mass spectrometry (GC–MS), and high-performance liquid chromatography (HPLC) were used to identify bioactive chemicals in the aqueous methanolic extracts of the acclimatized plants and compared with the donor plant. The bioactive chemicals were found considerably higher in the acclimatized plants (Phenols 95.0 mg GAE/g DW, flavonoids 99.2 mg QE/g DW, tannins 38.7 mg TAE/g DW, hesperidin 94.5 µg/mg, and quercetin 25.4 µg/mg). The antioxidant potential was also evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and demonstrated high scavenging activity at a concentration of 800 µg/mL in the acclimatized plants (83.0%). This study represents the first successful demonstration of in vitro regeneration, phytochemical profile analysis, and assessment of antioxidant activity in R. chalepensis plants established from synthetic seeds following short-term cold storage.

Acclimatization of Sugarcane Planet (Saccharum officinarum L.) From Synthetic Seeds In Several Media and Nutrition Concentrations

Benih sintetik tebu (Saccharum officinarum L.) merupakan embrio somatik yang dienkapsulasi dengan natrium alginat menjadi bentuk yang menyerupai benih. Tujuan penelitian ini untuk mengetahui kombinasi media tanam dan konsentrasi nutrisi yang tepat untuk aklimatisasi planlet tebu dari benih sintetik. Penelitian ini dilaksanakan pada bulan April 2019-Januari 2020 di Center of Development of Advance Science and Technology (CDAST) Universitas Jember. Rancangan penelitian dilakukan dengan menggunakan RAL faktorial dengan 2 faktor dan diulang sebangak 3 kali. Faktor pertama yaitu perlakuan media tanam arang sekam, perlite dan rockwool, sedangkan faktor kedua yaitu perlakuan konsentrasi nutrisi AB mix 500 ppm, 750 ppm, 1000 ppm dan 1250 ppm dan menggunakan analisis sidik ragam (ANOVA) apabila hasil yang diperoleh berbeda nyata maka dilakukan analisa lanjut dengan uji Duncan’s Multiple Range Test (DMRT) pada taraf kepercayaan 95%. Parameter yang diamati pada aklimatisasi adalah panjang akar, tinggi tanaman dan jumlah daun. Hasil aklimatisasi planlet dari benih sintetik menunjukkan bahwa media terbaik adalah rockwool dengan tinggi bibit 21,9 cm, jumlah daun 8 helai, panjang akar 13,6 cm. Konsentrasi nutrisi terbaik adalah AB mix 500 ppm dengan hasil tinggi tanaman 20,5 cm, jumlah daun 7,5 dan panjang akar 9,6 cm.

In vitro Conservation and Exploiting Polyembryonate Potential of Synthetic Seeds of Malaxis acuminata D. Don

Synthetic seeds were prepared in Malaxis acuminata by encapsulating shoot tip derived protocorm-like bodies (PLBs; 0.2-0.5 cm) in a hydrated gel capsule composed of calcium alginate matrix (sodium alginate; 3.5% and calcium chloride; 75 mM) in liquid Murashige and Skoog, 1962 (MS) medium. Synthetic seeds varied in physical features with the concentration of gelling agent and calcium chloride. In in vitro germination, the optimum response was achieved in MS medium fortified with BAP (1mgl-1); where synthetic seeds germinated with 100% frequency. Physically firm, globular, non-leaky, and self-breaking beads germinated efficiently which could be stored for up to 90 days at 4℃. At room temperature, synthetic seeds germinated with 50% frequency after 15 days and the germination frequency steadily reduced to 0% after 45 days of storage. Under in vivo conditions, synthetic seeds showed 80% bead-to-plant conversion in MS medium supplemented with antibacterial, antifungal agents and BAP (1mg/I). Plant Tissue Cult. & Biotech. 33(1): 9-15, 2023 (June)

In vitro cloning of stevia rebaudiana Bertoni to avoid verification and production of syn-thetic seeds

In vitro cloning of stevia rebaudiana Bertoni to avoid verification and production of syn-thetic seeds

Synthetic seeds for in vitro preservation of Asparagus officinalis L.

BackgroundAsparagus (Asparagus officinalis) is a perennial vegetable of economic importance for its high nutritional and medicinal value. Male plants are more desirable because of their higher spear yield. Therefore, the aim of this study was to evaluate the efficiency of the gibberellin inhibitors; paclobutrazol (PBZ) and cycocel (CCC) for in vitro preservation by encapsulation of in vitro-derived shoot tips of the superior germplasm of the first generation of asparagus male hybrid; cultivar Mary Washington 500W. MethodsAn efficient technique for synthetic seeds production was achieved, consisting of 3% (w/v) Na-alginate dissolved in water, Murashige and Skoog (MS) medium, MS medium with the growth inhibitors; PBZ and CCC at different concentrations, and 0.1 M calcium chloride (CaCl2.2H2O). Synthetic seeds were stored at 4 ℃ and then cultured after different storage durations (0, 2, 4, 8, 12, 16, 20 and 24 weeks) on MS medium supplemented with 1 mg l−1 kinetin, 0.2 mg l−1 naphthalene acetic acid and 0.5 mg l−1 gibberellic acid for shoot tip multiplication; then, recovered shoots were transferred to MS medium supplemented with 1 mg l−1 indole butyric acid and 0.5 mg l−1 PBZ for rooting.ResultsThe beads containing 1 mg l−1 PBZ considered the optimum for producing complete well-developed plantlets of Asparagus officinalis from recovered shoot tips after 24 weeks of storage that successfully acclimatized in the greenhouse.ConclusionsThis protocol is efficient for in vitro preservation by encapsulation of shoot tips and regeneration of Asparagus officinalis F1 male hybrid (Mary Washington 500w), using anti-gibberellin plant growth regulators within the beads to prolong the duration of storage and provide a continuous supply of the plant.

Short-Term Conservation of Juglans regia L. via Synthetic Seed Technology

Juglans regia L. is a crucial species as a forest tree and for its nutritional and medicinal values. It is also included in the list of endangered species in Albania and thus, there is a need to find methodologies to ensure its rapid regeneration and ex situ conservation. This research, investigated the regeneration of plantlets from synthetic seeds containing shoot tips of four native walnut varieties: ‘Përmet’, ‘Korçë’, ‘Peshkopi’, and ‘Tropojë’. First, in vitro-derived shoot tips from walnut seedlings are encapsulated using sodium alginate. After that, the regeneration potential of the encapsulated shoot tips and the influence of incubation conditions are evaluated. The synthetic seeds were incubated at either 25 °C or 8 °C, with and without dehydration treatment, in 0.5 M sucrose solution for 3 h. The synthetic seeds in both temperature regimes (25 °C and 8 °C) develop plantlets and provid conservation potential without the need for subcultures for 4 and 3.5 months, respectively. Furthermore, all walnut varieties incubated in these conditions achiev a high regeneration rates.

Comparative carbon footprints of buffalo milk produced at smallholder and organised farms in Hisar district of Haryana, India

The present study was conducted to compare the carbon footprint of milk produced in 75 smallholder farms and two organised buffalo farms in Hisar district of Haryana using Life Cycle Assessment approach. Primary data was collected from farmers and farm managers for the study. Functional unit was one litre of milk with system boundary being ‘Cradle-to-farm gate’. Methodology prescribed by Intergovernmental Panel on Climate Change was used for estimation of emissions from different sources. Secondary data was also relied upon for synthetic fertilizers and seeds, combustion of fossil fuel, production of concentrates and dry fodders. The average carbon footprint of milk produced in rural smallholder and organised farms were 3.54 and 4.53 kg CO₂-eq./L milk, respectively, indicating superiority of village level production systems. Methane from enteric fermentation was estimated to be contributing nearly half of the total greenhouse gas emissions. It is suggested that rural smallholder production systems should be favoured over organised ones given the lower greenhouse gas emissions.

Role of Biotechnology in Improving Crop Yield and Enhancing Food Security

Biotechnology has had a huge impact on agriculture and ultimately enhanced our quality of life by studying, altering, and cloning various crops. Hybrid seeds, synthetic seeds, photosynthesis boosters, stress-resistant plants and crops, bio-fertilizers, and bio-pesticides are a few examples of potential applications. Biotechnology may be useful for managing livestock, preserving agricultural goods, and sustaining present crop yields while using fewer pesticides, herbicides, and fertilisers. In addition to improving traditional plant breeding techniques, biotechnology also provides a better replacement for food that is produced. When used in conjunction with other cutting-edge agricultural technologies, it offers a creative and environmentally friendly solution to meet consumer demand for sustainable agriculture. Biotechnology significantly increases agricultural output by enhancing disease prevention and drought and flood resistance. This not only satisfies the rising need for food but also aids in the financial success of farmers. Techniques based on biotechnology provide effective, cheap solutions to pest control problems. Producers have created a protein that effectively eliminates pest issues with cotton, maize, and potato crops.

Micropropagation of Pterocarpus marsupium Roxb. through synthetic seeds and its novel antibiofilm activities against ESKAPE pathogens

Pterocarpus marsupium, an endangered medicinal tree, desists long distance transportation of its recalcitrant propagules, eclipsing the mass vegetative propagation and hindering the pharmaceutical application. Therefore, the present study attempted to develop synthetic seeds by encapsulating the nodal segments using sodium alginate and calcium chloride solutions. A concentration of 100 mM calcium chloride and 3 % sodium alginate proved optimum for preparing uniform synthetic seeds. The highest 76.7 % conversion response of synthetic seed into 8.16 shoots with mean shoot length (4.35 cm) was obtained on Murashige and Skoog (MS) medium containing meta-topolin (5.0 µM) and α-naphthalene acetic acid (1.0 µM) after 12 weeks of culture. However, the shoots failed to induce a rooting response on the aforementioned optimized medium. Therefore, a two-step in vitro rooting procedure was developed. Synthetic seeds of this plant retained their germination potential after storage at 4 ᵒC for eight weeks. Analysis of glutathione-S-transferase (GST) activity indicated that the in vitro culture of synthetic seed on the above medium experienced the lowest physiological stress. The regenerated plantlets were successfully acclimatized to the natural environment, where they showed more than a 90 % survival rate after three months. The heartwood aqueous extract of the tree displayed novel antibiofilm activities against multi-drug resistant ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobcter spp.) pathogens. This study facilitates the multiplication, conservation, and production of novel bioactive compounds.