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The instability of in vitro cultures may cause genetic and epigenetic changes in crops called somaclonal variations. Sometimes, these changes produce beneficial effects; for example, they can be used in breeding programs to generate new cultivars with desirable characteristics. In this article, we present a systematic review designed to answer the following question: How does somaclonal variation contribute to plant genetic improvement? Five electronic databases were searched for articles based on pre-established inclusion and exclusion criteria and with a standardized search string. The somaclonal variation technique has been most frequently applied to ornamental plants, with 49 species cited in 48 articles, and to the main agricultural crops, including sugarcane, rice, banana, potato and wheat, in different countries worldwide. In 69 studies, a technique was applied to evaluate the genetic diversity generated between clones, and, in 63 studies, agronomic performance characteristics were evaluated. Other studies are related to resistance to pathogens, ornamental characteristics and resistance to abiotic stresses. The application of the plant growth regulators (PGRs) benzylaminopurine (BAP) and dichlorophenoxyacetic acid (2,4-D) was the most common method for generating somaclones, and randomly amplified polymorphic DNA (RAPD) molecular markers were the most commonly used markers for identification and characterization. Somaclonal variation has been used in genetic improvement programs for the most economically important crops in the world, generating genetic diversity and supporting the launch of new genotypes resistant to diseases, pests and abiotic stresses. However, much remains to be explored, such as the genetic and epigenetic mechanisms from which somaclonal variation is derived.

Agricultural crops are exposed to various abiotic stresses, such as salinity, water deficits, temperature extremes, floods, radiation, and metal toxicity. To overcome these challenges, breeding programs seek to improve methods and techniques. Gene editing by Clustered Regularly Interspaced Short Palindromic Repeats—CRISPR/Cas—is a versatile tool for editing in all layers of the central dogma with focus on the development of cultivars of plants resistant or tolerant to multiple biotic or abiotic stresses. This systematic review (SR) brings new contributions to the study of the use of CRISPR/Cas in gene editing for tolerance to abiotic stress in plants. Articles deposited in different electronic databases, using a search string and predefined inclusion and exclusion criteria, were evaluated. This SR demonstrates that the CRISPR/Cas system has been applied to several plant species to promote tolerance to the main abiotic stresses. Among the most studied crops are rice and Arabidopsis thaliana, an important staple food for the population, and a model plant in genetics/biotechnology, respectively, and more recently tomato, whose number of studies has increased since 2021. Most studies were conducted in Asia, specifically in China. The Cas9 enzyme is used in most articles, and only Cas12a is used as an additional gene editing tool in plants. Ribonucleoproteins (RNPs) have emerged as a DNA-free strategy for genome editing without exogenous DNA. This SR also identifies several genes edited by CRISPR/Cas, and it also shows that plant responses to stress factors are mediated by many complex-signaling pathways. In addition, the quality of the articles included in this SR was validated by a risk of bias analysis. The information gathered in this SR helps to understand the current state of CRISPR/Cas in the editing of genes and noncoding sequences, which plays a key role in the regulation of various biological processes and the tolerance to multiple abiotic stresses, with potential for use in plant genetic improvement programs.

The fungus Fusarium oxysporum f. sp. cubense (FOC), tropical race 4 (TR4), causes Fusarium wilt of banana, a pandemic that has threatened the cultivation and export trade of this fruit. This article presents the first systematic review of studies conducted in the last 10 years on the resistance of Musa spp. to Fusarium wilt. We evaluated articles deposited in different academic databases, using a standardized search string and predefined inclusion and exclusion criteria. We note that the information on the sequencing of the Musa sp. genome is certainly a source for obtaining resistant cultivars, mainly by evaluating the banana transcriptome data after infection with FOC. We also showed that there are sources of resistance to FOC race 1 (R1) and FOC TR4 in banana germplasms and that these data are the basis for obtaining resistant cultivars, although the published data are still scarce. In contrast, the transgenics approach has been adopted frequently. We propose harmonizing methods and protocols to facilitate the comparison of information obtained in different research centers and efforts based on global cooperation to cope with the disease. Thus, we offer here a contribution that may facilitate and direct research towards the production of banana resistant to FOC.

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) is the most limiting factor in the banana agribusiness worldwide. Therefore, studies regarding pathogen attack mechanisms, and especially host defense responses, in this pathosystem are of utmost importance for genetic breeding programs in the development of Foc-resistant banana cultivars. In this study, analysis at the molecular, histological and histochemical levels of the Musa spp. x Foc interaction was performed. Three Foc isolates representative of race 1 (R1), subtropical race 4 (ST4) and isolate 229A, which is a putative ST4, were inoculated in two Prata-type cultivars (Prata-Anã and BRS Platina) and one cultivar of the Cavendish type (Grand Naine). Of seven genes related to plant–pathogen interactions, five were overexpressed in ‘BRS Platina’ 12 h after inoculation (HAI) with Foc R1 and ST4 but had reduced or negative expression after inoculation with Foc 229A, according to RT–qPCR analyses. While hyphae, mycelia and spores of the Foc 229A isolate grow towards the central cylinder of the Grand Naine and Prata-Anã cultivars, culminating in the occlusion of the xylem vessels, the BRS Platina cultivar responds with increased presence of cellulose, phenolic compounds and calcium oxalate crystals, reducing colonization within 30 days after inoculation (DAI). In general, these data indicate that the cultivar BRS Platina has potential for use in banana-breeding programs focused on resistance to Foc tropical race 4 (TR4) and in aggregating information on the virulence relationships of the Foc pathogen and the defense responses of banana plants after infection.

Phytoene desaturase (PDS) is a plant enzyme involved in carotenoid biosynthesis. The PDS gene has been used as a selective marker for genome editing in several plant species, including banana (Musa spp.). Its knockout promotes dwarfism and albinism, characteristics that are easily recognizable and highly favorable. In Musa spp., the A genome increases fruit production and quality, whereas the B genome is associated with tolerance to biotic and abiotic stresses. The objective of this study was to identify a molecular marker in the PDS gene to easily discriminate the A and B genomes of banana. A 2166 bp fragment for the “PDSMa” marker was identified as polymorphic for the A genome (identification accuracy of 99.33%), whereas ~332 and ~225 bp fragments were detected for the “PDSMb” marker with 100% accuracy using MedCalc software. In this study, we used genotypes with A and B genomes that are used in the genetic improvement of bananas and an accession with the BT genome. It was not possible to differentiate the accession with the BT genome from the others, suggesting that the markers do not have the capacity to separate the T genome from the A and B genomes. To the best of our knowledge, this is the first study to use the PDS gene to determine doses of the A genome and identify the B genome in Musa spp., which will aid in evaluating the genomic constitution of banana hybrids and accessions at the seedling stage and accelerating their classification in crop genetic improvement programs.

Bananas are the main fruits responsible for feeding more than 500 million people in tropical and subtropical countries. Black Sigatoka, caused by the fungus Pseudocercospora fijiensis, is one of the most destructive disease for the crop. This fungus is mainly controlled with the use of fungicides; however, in addition to being harmful to human health, they are associated with a high cost. The development of resistant cultivars through crosses of susceptible commercial cultivars is one of the main focuses of banana breeding programs worldwide. Thus, the objective of the present study was to investigate the interaction between Musa sp. and P. fijiensis through the relative expression of candidate genes involved in the defence response to black Sigatoka in four contrasting genotypes (resistant: Calcutta 4 and Krasan Saichon; susceptible: Grand Naine and Akondro Mainty) using quantitative real-time PCR (RT–qPCR) in addition to histological and histochemical analyses to verify the defence mechanisms activated during the interaction. Differentially expressed genes (DEGs) related to the jasmonic acid and ethylene signalling pathway, GDSL-like lipases and pathogenesis-related proteins (PR-4), were identified. The number and distance between stomata were directly related to the resistance/susceptibility of each genotype. Histochemical tests showed the production of phenolic compounds and callosis as defence mechanisms activated by the resistant genotypes during the interaction process. Scanning electron microscopy (SEM) showed pathogenic structures on the leaf surface in addition to calcium oxalate crystals. The resistant genotype Krasan Saichon stood out in the analyses and has potential for use in breeding programs for resistance to black Sigatoka in banana and plantains.

Yellow Sigatoka, caused by the fungus Pseudocercospora musae, represents one of the most challenging diseases in bananas, which is aggravated due to the genetic variability of this pathogen. The main objective of this study was to characterize the infection process of P. musae in two banana cultivars with different levels of resistance and to quantify the expression of resistance genes. Inoculated and non-inoculated leaf samples of the two cultivars, Yangambi km 5—resistant and Grande Naine—susceptible, were investigated by light microscopy, histochemistry, scanning electron microscopy, and RT-qPCR. The cultivar Grande Naine showed early signs of P. musae infection, including the production of lipophilic substances and phenolic compounds, while the cultivar Yangambi km 5 was not affected. In the Grande Naine cultivar, the protein kinase gene was upregulated, while in the Yangambi km 5, it was repressed. The WAK2 gene was initially upregulated in both cultivars; however, in Grande Naine, it was subsequently repressed, while in Yangambi km 5, it remained downregulated. These findings have the potential to contribute to more effective management strategies in the control of yellow Sigatoka and the development of banana varieties resistant to yellow Sigatoka.

Fusarium wilt is a soil borne fungal disease that has devastated banana production in plantations around the world. Most Cavendish-type bananas are susceptible to strains of Fusarium oxysporum f. sp. cubense (Foc) belonging to the Subtropical Race 4 (STR4) and Tropical Race 4 (TR4). The wild banana diploid Musa acuminata ssp. malaccensis (AA, 2n = 22) carries resistance to Foc TR4. A previous study using segregating populations derived from M. acuminata ssp. malaccensis identified a quantitative trait locus (QTL) (12.9 cM) on the distal part of the long arm of chromosome 3, conferring resistance to both Foc TR4 and STR4. An SNP marker, based on the gene Macma4_03_g32560 of the reference genome ‘DH-Pahang’ v4, detected the segregation of resistance to Foc STR4 and TR4 at this locus. Using this marker, we assessed putative TR4 resistance sources in 123 accessions from the breeding program in Brazil, which houses one of the largest germplasm collections of Musa spp. in the world. The resistance marker allele was detected in a number of accessions, including improved diploids and commercial cultivars. Sequencing further confirmed the identity of the SNP at this locus. Results from the marker screening will assist in developing strategies for pre-breeding Foc TR4-resistant bananas. This study represents the first-ever report of marker-assisted screening in a comprehensive collection of banana accessions in South America. Accessions carrying the resistance marker allele will be validated in the field to confirm Foc TR4 resistance.

The objective of this systematic review (SR) was to select studies on the activity of glycerol as a molecule that induces disease resistance in plants. We sought to evaluate articles deposited in five electronic databases using a search string and predefined inclusion and exclusion criteria. The most studied crops are Arabidopsis thaliana, Glycine max, and Coffea spp. The most commonly cited biotic agents include Pseudomonas syringae, Blumeria graminis, and Colletotrichum higginsianum. Numerous doses of glycerol were studied, and concentrations ranged from 0.004 to 9.21%, with a 3% concentration of glycerol being considered most effective for most plant species, where greater resistance was observed with increased glycerol-3-phosphate (G3P) and decreased oleic acid levels. The main means of application of the product were spraying and immersion. The SR also revealed the evaluation of resistance-inducing genes, such as PR proteins (PR-1, PR2, PR-5, etc.), HPS70, HSP90, SCAM4, and Tapr1, among others. The information collected in this SR helps to understand the state of the art on the use of glycerol as a molecule inducing resistance against biotic stressors to understand the mechanisms involved in most host–pathogen relationships. This information will be useful in plant breeding programs and for growers/producers.

Drought stress is one of the main environmental factors that limit plant growth and productivity. In order to endure such stress, which increases every year, plants developed different strategies to withstand drought conditions, including an enzymatic antioxidant defense system with enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxidase (POX), glutathione peroxidase (GPX), and glutathione-S-transferase (GST), among others, playing key roles. Therefore, efforts to develop new drought-tolerant varieties have become essential for maintaining human populations increasing researchers’ interest in tools and/or mechanisms involved in drought tolerance. Thus, this work is aimed at offering a systematic review of the correlation between gene expression and antioxidant enzyme activity in water-stress tolerant plants for fourteen most important food crops worldwide, according to FAO, and Arabidopsis thaliana. This systematic review was performed using the free software StArt and the PICOS strategy. Among the factors that generate drought tolerance is the increased activity/gene expression of antioxidant enzymes. The use of plant regulators stood out as the most critical substance among chemical inputs; however, these do not provide long-term tolerance and therefore, the development of genetically modified organisms would be the most efficient technique for promoting tolerance to long-term water deficit in plants. In this systematic review, data was collected, and forty candidate genes were identified as responsible for water deficit tolerance, which can be used by plant breeding programs to develop genetically engineered products.