Haploids, containing a single set of chromosomes (n), allow direct gene effect observation, while doubled haploids, produced by chromosome doubling, are fully homozygous and accelerate plant genetic improvement. Dihaploids, derived from tetraploid species, aid genetic studies and polyploid crop enhancement. This mini-review examines ten plant species improved using these techniques, five biological limitations affecting their application, and the impact of doubled haploid variation on barley genetic progress. Barley cultivation in the Peruvian highlands has faced performance challenges, leading researchers to employ the doubled-haploid technique for better-adapted varieties. Three barley crosses—Ya/LM94, B16/LM94, and B12/LM94—generated doubled-haploid lines evaluated in Acobamba alongside parental and commercial controls. The Ya/LM94-PC27 line closely matched the expert-defined ideal, offering higher yield, shorter plant height, stripe rust resistance, and optimal grain quality, while reducing research costs by 26

Identifying physiological traits that respond to stress is a challenge in plant biotechnology, given the complexity of salinity and water-deficit effects. This study evaluated the usefulness of Principal Component Analysis (PCA) to highlight the most relevant indicators in sugarcane and pineapple cultured in temporary immersion bioreactors (TIBs). Secondary data from previous experiments were used, including shoot multiplication rate, fresh biomass, and biochemical profiles (aldehydes, chlorophylls, carotenoids, and phenolics). PCA reduced dataset dimensionality and emphasized the variables with the greatest contribution to experimental variance, namely soluble phenolics, malondialdehyde, other aldehydes, and cell wall-linked phenolics. These findings confirmed that PCA is a valuable tool to guide breeding programs toward the selection of stress-resilient genotypes.

Cryopreservation using liquid nitrogen (LN) is a widely adopted technique for the long-term conservation of plant genetic resources. While effective at preserving seed viability, its effects on early plant development remain insufficiently characterized, particularly in critical crops like maize. This study evaluated the early growth responses of maize seedlings originating from seeds subjected to LN exposure, focusing on seedling emergence, root system morphology, and biomass partitioning among plant organs. Maize seeds (cv Tuzón) were dried to 6

Artificial intelligence (AI) is transforming plant science by enabling rapid data analysis, predictive modeling, and precision breeding. Image recognition accelerates phenotyping, while machine learning optimizes bioprocesses, improving both reproducibility and scalability. Deep Learning technologies, and cloud services expand accessibility, with Copilot exemplifying AI’s role by documenting leaf dehydration in Phaseolus vulgaris L. under controlled conditions. Statistical analysis confirmed a significant weight reduction from 0.25 g to 0.13 g (52%). Copilot generated structured annotations of morphological changes, including loss of turgor, increased venation prominence, and surface wrinkling. Mathematical overlays revealed fractal branching and Voronoi tessellation patterns, with dehydration exaggerating vein relief and sharpening boundaries, linking morphological traits to physiological stress responses. Additionally, Copilot supported bilingual figure legends, terminology harmonization, and bibliometric keyword suggestions, streamlining reproducibility and clarity. Overall, this AI-human synergy demonstrates Copilot’s value as a methodological partner, enhancing accessibility, precision, and international impact in plant physiology research.

Capsicum annuum L. comprises several cultivars, including True Heart, whose fruit quality is influenced by the organic matter incorporated into the substrate. This study evaluated the fruits of C. annuum (True Heart cultivar) grown in Red Ferralitic Soil from Ciego de Ávila, Cuba, amended with organic matter derived from the invasive aquatic plants Pistia stratiotes L. and Pontederia crassipes Mart. Twenty-five experimental units were established: five with a simple substrate (Red Ferralitic Soil), five units each with mixed substrates containing 0.5

Pineapple (Ananas comosus L. Merr., cultivar MD2) is a key tropical crop but highly sensitive to salinity and drought, making temporary immersion bioreactors (TIBs) valuable for micropropagation and stress studies. Apical buds were cultured in vitro, transferred to TIBs with paclobutrazol, and subjected to nine treatments (control, NaCl, mannitol at 50.0 to 200.0 mM) for 30 d. Growth and biochemical traits were measured, data normalized (0 to 1), and analyzed by hierarchical cluster analysis (HCA) using average linkage and squared Euclidean distance. The dendrogram revealed clear treatment similarities: controls remained distinct until higher rescaled distances, low concentrations clustered together, and high concentrations formed separate groups, confirming dose-dependent responses. NaCl and mannitol did not cluster together initially, indicating distinct physiological profiles, with NaCl imposing additional ionic toxicity beyond osmotic effects. These findings complement earlier evaluations showing significant reductions in shoot multiplication and biomass above 50.0 mM, with critical concentrations for 50

Sugarcane is of global economic significance, serving as a primary source of sugar, bioethanol, and feedstock for diverse agro – industrial applications. However, its productivity is frequently constrained by abiotic stressors, notably salinity and drought. This study employed four multivariate regression models derived from previously published in vitro datasets to quantify the impacts of stress agents (NaCl and mannitol) and key biochemical markers — malondialdehyde, assorted aldehydes, chlorophylls a and b, carotenoids, soluble phenolics, and cell wall – bound phenolics—on shoot regeneration and biomass accumulation. Cultures were maintained in temporary immersion bioreactors under standardized conditions. Model performance was robust across stress treatments (adjusted R²: 0.8864–0.9887). Under NaCl stress, shoot cluster fresh weight increased with other aldehydes (+ 11.14) and soluble phenolics (+ 3.95), while malondialdehyde (− 8.49) and carotenoids (− 4.09) reduced biomass. Shoot multiplication responded positively to malondialdehyde (+ 3.80) and negatively to aldehydes (− 4.12). Under mannitol stress, all parameters were classified as low values. The biochemical landscape suggests distinct functional roles – malondialdehyde as a stress signal, aldehydes as cytotoxic agents, phenolics as protectants, and carotenoids functioning as stress-responsive compounds. These findings underscore a biochemical trade – off between defense and regeneration, with implications for optimizing stress – conditioned micropropagation systems and leveraging biochemical predictors for enhanced sugarcane propagation. Sugarcane productivity under salinity and drought reflects a biochemical trade-off, where aldehydes, phenolics, carotenoids, and malondialdehyde shape regeneration and biomass, guiding stress-conditioned micropropagation strategies.

Hierarchical Cluster Analysis (HCA) is a technique for grouping objects based on their similarities, forming a hierarchical structure represented as a dendrogram. It follows two main strategies: agglomerative (bottom-up) and divisive (top-down). HCA is extensively applied across disciplines such as biology, marketing, and social sciences. In plant biotechnology, it plays a crucial role in assessing genetic diversity, phenotypic variations, metabolomic patterns, pathogen resistance, and environmental adaptability. Although HCA is not widely employed in plant germplasm cryopreservation research, it holds potential for evaluating the genetic diversity and stability of cryopreserved specimens. This communication illustrates the novel application of HCA in a common bean seed cryopreservation experiment. This study investigated the effects of cryopreservation on P. vulgaris seeds during early germination. No visible phenotypic changes were seen in seedlings from cryopreserved seeds. However, significant biochemical changes were noted: cryopreservation increased aldehydes (excluding malondialdehyde) in shoots from 56.47 to 253.19 µmol g− 1 fresh weight, and reduced phenolics in roots. Cut line 1 in the dendrogram identified two main groups, with leaves distinctly different from other organs. Cut line 2 separated non-cryopreserved roots in branch 1 and cryopreserved roots in branch 2. HCA, as a methodological tool, confirmed roots were more affected by cryostorage.

Sugarcane is a globally important crop cultivated in over 100 countries, serving as a primary source of sugar and bioethanol while supporting rural economies and sustainable agriculture through its high biomass yield, adaptability, and ongoing genetic improvement for climate resilience. As climate change intensifies drought and salinity stress in key production regions, breeding stress-tolerant cultivars has become essential. This study applied Euclidean distance metrics for simultaneous trait selection, evaluating six in vitro culture–derived, potentially mutant, sugarcane plantlets cultivated in planting beds. Morphological traits, such as shoot height, stem diameter, and tillering capacity, along with biochemical indicators, including chlorophyll content, soluble phenolics, and aldehyde accumulation, revealed substantial variation and differential stress responses. Standardization and Euclidean analysis quantified each plantlet’s divergence from expert-defined ideal profiles, identifying genotypes with agronomic and biochemical indicators closely aligned with the target values. These findings demonstrated the effectiveness of multivariate selection in sugarcane improvement, with Euclidean metrics providing a holistic framework for ranking candidates and guiding future field validation under climate-challenged conditions. Plantlet 11 exhibited the highest Euclidean distance (2.23) from the expert breeder’s reference profile, indicating the greatest deviation from the target indicators. Conversely, plantlet 12 achieved the most favorable integral evaluation, with the lowest observed Euclidean distance (1.15), reflecting the closest alignment with the desired traits.

Maintaining one difference between experimental groups ensures valid results, ties changes to treatments, enhances credibility, and strengthens research quality through consistency. Exceptions, however, can be acceptable in specific circumstances.

Nanoparticles, typically from 1 to 100 nm in size, have unique physical and chemical properties that allow them to penetrate biological barriers, making them effective in plant biotechnology for delivering genes, nutrients, and pesticides. Absorbed through roots, leaves, or seeds, they move through the vascular system, enhancing agricultural practices and productivity. Their applications include gene delivery with gold nanoparticles, nanofertilizers for efficient nutrient delivery, and silver nanoparticles for effective pest control, representing significant advancements in sustainable agriculture. However, challenges such as potential toxicity to plants and non-target organisms, high production costs, and environmental impact are the uncertainties that require further research for safe and widespread adoption. Emerging trends include smart nanocarriers that respond to environmental triggers and biodegradable nanoparticles to minimize environmental accumulation. Future research aims to enhance delivery precision and integrate nanotechnology with tools like CRISPR-Cas9 and machine learning for advanced crop management, potentially revolutionizing plant biotechnology and boosting global food security. A study on Stevia rebaudiana revealed that using silver nanoparticles (AgNPs: Argovit™) in temporary immersion bioreactors (TIBs) improved propagation and stimulated secondary metabolite production. Specifically, 25.0 and 37.5 mg L−1 AgNPs reduced shoot multiplication, while 12.5 mg L−1 enhanced endogenous diterpene levels. Similarly, applying AgNPs in TIBs to Gerbera jamesonii increased vase life by 21

Temporary immersion bioreactors (TIBs) have revolutionized plant micropropagation by providing an efficient, cost-effective alternative to traditional methods. By periodically immersing plant tissues in liquid culture media, TIBs optimize nutrient absorption and gas exchange, significantly enhancing growth rates and plant quality, particularly for commercially and ecologically valuable species such as Aristotelia chilensis, Coffea arabica, and Musa spp. Recent advancements, including refined immersion protocols, have further improved plant development while reducing production costs, demonstrating TIBs’ adaptability for sustainable agriculture. Beyond micropropagation, TIBs support secondary metabolite production, and conservation of endangered species, expanding their role in biotechnology. However, challenges such as high setup costs, complex maintenance, and species-specific limitations suggest areas for continued improvement. The versatility of TIBs is evident in their applications for sugarcane and potato cultivation. A two-step protocol for sugarcane, combining paclobutrazol and gibberellic acid, has doubled multiplication rates and reduced production costs by 46

Drought stress hinders plant development by limiting water availability, disrupting osmotic equilibrium, and impairing nutrient uptake and photosynthetic efficiency. To counter these effects, temporary immersion bioreactors (TIBs) provide a scalable platform for rapid shoot proliferation and in vitro screening of sugarcane genotypes subjected to mannitol-induced osmotic stress. These controlled responses often parallel field performance, enabling efficient early-stage selection. In this study, apical meristems of sugarcane (cultivar C-1051-73) were cultured in vitro, transferred to TIBs, and exposed to mannitol concentrations ranging from 0 to 200 mM. Key physiological parameters and biochemical stress markers were quantified. Discriminant analysis, implemented via Python-based machine learning, classified stress responses using Fisher’s linear function. Control (0 mM) and high-stress (200 mM) treatments served as training sets, while intermediate concentrations (50–150 mM) were used for model validation. The analysis identified 150 mM as the threshold for stress manifestation, with lower concentrations classified as non-stressed. This analytical framework strengthens genetic improvement strategies—including selection, hybridization, mutagenesis, and transgenesis—by enabling early identification of drought-tolerant genotypes prior to field deployment. The approach reduces costs, shortens breeding cycles, and enhances decision-making in cultivar development. Moreover, the methodology is transferable to other crops, provided that discriminant functions are recalibrated to accommodate species-specific physiological profiles. Altogether, the integration of TIBs with discriminant analysis offers a robust, cost-efficient solution for drought tolerance screening, advancing precision agriculture and promoting crop resilience under water-limited conditions. Temporary immersion bioreactors combined with discriminant analysis enable fast, economical in vitro screening of sugarcane genotypes under osmotic stress—accelerating drought tolerance selection and supporting scalable crop improvement.

Natural plant metabolites, also known as secondary metabolites, are crucial for the survival and adaptation of plants in their environment. For modern society, these metabolites can be produced under uncontrolled or controlled environments. Alkaloids, flavonoids, terpenoids, phenols, lignans, saponins, coumarins, and glucosinolates are among the most important natural products from plants. Proteases from pineapple cultures in temporary immersion bioreactors have been obtained. Additionally, the effects of sodium chloride, mannitol, and sodium azide on pineapple shoots propagated in temporary immersion bioreactors were recorded, with levels of chlorophylls, carotenoids, aldehydes, and phenolics measured. This short review summarizes the main results obtained.

The chi-square test (χ2) has long been used to analyze categorical data and, while not used extensively in biotechnology trials, it has found application in numerous studies investigating segregation analysis. This report considered whether this test can be used on quantitative data collected from biotechnology experiments to determine whether the inherent principles of the test would allow room for adaptation and application with numerical data. This was evaluated using data published in three diverse biotechnology experiments investigating the response of in vitro sugarcane plants to salt stress, the effect of genetic transformation on field performance of pineapple plants, and the effect of cryopreservation on neonotonia seeds. Overall, this work shows that the χ2 test is suitable for the analysis of quantitative (numerical) data.

Sugarcane is a vital crop in tropical and subtropical regions worldwide, contributing to economic growth, energy production, and nutritional value. Over the past decade, numerous studies have demonstrated that in vitro screening for stress tolerance enables rapid evaluation of many plants, propagules, and explants for desirable traits, significantly accelerating the breeding process compared to traditional field methods. This report showcases the correlation between responses to NaCl (salinity) and mannitol (drought) in sugarcane plantlets micropropagated in temporary immersion bioreactors. With coefficients of determination (R2) exceeding 90

Pineapple is a highly valuable tropical fruit crop with significant economic importance. In pineapple cultivation, several traits could benefit from genetic improvement, such as disease resistance, drought tolerance, fruit quality, shelf life, and yield. The methods used to genetically improve pineapple include traditional breeding, mutagenesis, biotechnology, genetic engineering, and in vitro culture. A few notable scientific groups worldwide performing pineapple genetic transformation include the South China Agricultural University, the Chinese Academy of Tropical Agricultural Sciences, the University of Hawaii, and Del Monte. The main transgenes introduced into the pineapple genome are the antisense ACC oxidase gene, herbicide resistance genes, carotenoid biosynthesis genes, and disease resistance genes. Our group at the Bioplant Centre, University of Ciego de Ávila, Cuba, also transformed pineapple and evaluated the field performance over more than 8 years. We compared transgenic ¨Cayena Lisa Serrana¨ cultivar plants to macropropagated controls, evaluating physiological and biochemical indicators of the phenotype. Out of 160 characters evaluated, significant differences were observed in 88 characters between the transgenic plants and the conventionally vegetative propagated plants, and in 70 characters between the transgenic plants and the micropropagated plants. Overall, the phenotype of the transformed material conforms to the phenotype of the two controls. These consistent data reflect the usefulness of characterizing the modified plants.

Lilium spp., valued worldwide for their ornamental and medicinal properties, benefit from in vitro propagation techniques enhanced by silver nanoparticles (AgNPs). This study evaluated how different concentrations of Argovit™ AgNPs (0 to 100.0 mg•L⁻1) affect the growth and acclimatization of Lilium ‘Concador select’ micro-bulbs. Plants were cultured in modified Murashige and Skoog medium and later acclimatized in peat-vermiculite trays across two research sites in Saltillo, Mexico. Key morphological traits were measured after transplant. Statistical modeling revealed that bulb diameter responded most consistently to AgNP treatment, making it a reliable indicator of effectiveness. Cluster analysis grouped nanoparticle doses into three categories, showing that moderate concentrations promoted uniform growth, while very low or high doses led to variable outcomes. These findings supported the targeted use of AgNPs to improve micropropagation efficiency and informed breeding and conservation strategies for Lilium.

Plant genetic improvement integrates conventional breeding with advanced biotechnological approaches to enhance traits such as yield, disease resistance, and stress tolerance. Among these, in vitro-induced somaclonal variation—genetic and epi-genetic alterations arising during tissue culture—has emerged as a valuable tool for crop improvement. This variation can lead to novel phenotypes suitable for selection and propagation. Recent studies have demonstrated its utility in crops such as sugarcane, rice, banana, potato, wheat, tomato, barley, chrysanthemum, soybean, and maize. This review distinguishes itself by providing the first integrated evaluation of somaclonal variation applications across major crops alongside a detailed case study of pineapple, a species seldom emphasised in prior reviews. As one of the most widely cultivated tropical fruits with significant commercial value in both fresh and processed markets, pineapple plays a vital role in the agricultural economies of many developing countries. We highlight results from somaclonal variants derived from the Red Spanish cultivar, including P3R5 and Dwarf, which exhibited significant morphological and physiological differences. Amplified Fragment Length Polymorphism confirmed genetic divergence, with Dwarf showing enhanced water-use efficiency and antioxidant activity. These findings underscore somaclonal variation’s potential as a complementary strategy to conventional breeding, contributing to crop diversification and agricultural resilience.

Sugarcane and pineapple are vital crops with significant economic contributions to agriculture, necessitating enhancements in their genetic traits to improve sustainability and profitability. Sodium azide mutagenesis serves as a valuable tool in plant breeding and genetic studies, offering opportunities to explore induced physiological responses for safer and more effective applications. This study examines the effects of four sodium azide concentrations (0, 0.15, 0.30, and 0.45 mM) on sugarcane and pineapple cultivated in temporary immersion bioreactors (TIBs), employing principal component analysis (PCA) for detailed trait evaluation. PCA, a robust statistical method, identified malondialdehyde, chlorophyll b, and cell wall–linked phenolics as key indicators contributing to variance explanation. Comparing the findings, sugarcane results underscore a trade-off between stress markers (malondialdehyde, carotenoids, and soluble phenolics) and growth traits (fresh shoot mass), revealing the impact of stress responses on biomass. Meanwhile, pineapple outcomes emphasize a trade-off between photosynthetic pigments (chlorophyll a, chlorophyll b, carotenoids) and growth traits (shoot multiplication rate and fresh shoot mass), highlighting a shift toward photosynthetic efficiency over vegetative propagation. These contrasting principal components reflect how sugarcane adapts to stress while pineapple optimizes photosynthesis under varying conditions. Together, the analyses showcase how different species prioritize physiological traits based on their unique developmental or environmental demands, offering insights into tailored approaches for crop improvement and genetic research. This integrated study contributes valuable knowledge for advancing agricultural sustainability and productivity in these economically essential crops.