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Banana (Musa spp.), which is one of the world’s most popular and most traded fruits, is highly susceptible to pests and diseases. Pseudocercospora musae, responsible for Sigatoka leaf spot disease, is a principal fungal pathogen of Musa spp., resulting in serious economic damage to cultivars in the Cavendish subgroup. The aim of this study was to characterize genetic components of the early immune response to P. musae in Musa acuminata subsp. burmannicoides, var. Calcutta 4, a resistant wild diploid. Leaf RNA samples were extracted from Calcutta 4 three days after inoculation with fungal conidiospores, with paired-end sequencing conducted in inoculated and non-inoculated controls using lllumina HiSeq 4000 technology. Following mapping to the reference M. acuminata ssp. malaccensis var. Pahang genome, differentially expressed genes (DEGs) were identified and expression representation analyzed on the basis of gene ontology enrichment, Kyoto Encyclopedia of Genes and Genomes orthology and MapMan pathway analysis. Sequence data mapped to 29,757 gene transcript models in the reference Musa genome. A total of 1073 DEGs were identified in pathogen-inoculated cDNA libraries, in comparison to non-inoculated controls, with 32% overexpressed. GO enrichment analysis revealed common assignment to terms that included chitin binding, chitinase activity, pattern binding, oxidoreductase activity and transcription factor (TF) activity. Allocation to KEGG pathways revealed DEGs associated with environmental information processing, signaling, biosynthesis of secondary metabolites, and metabolism of terpenoids and polyketides. With 144 up-regulated DEGs potentially involved in biotic stress response pathways, including genes involved in cell wall reinforcement, PTI responses, TF regulation, phytohormone signaling and secondary metabolism, data demonstrated diverse early-stage defense responses to P. musae. With increased understanding of the defense responses occurring during the incompatible interaction in resistant Calcutta 4, these data are appropriate for the development of effective disease management approaches based on genetic improvement through introgression of candidate genes in superior cultivars.

Endoparasitic root-knot nematodes (RKNs) ( Meloidogyne spp.) cause considerable losses in banana ( Musa spp.), with Meloidogyne incognita a predominant species in Cavendish sub-group bananas. This study investigates the root transcriptome in Musa acuminata genotypes 4297-06 (AA) and Cavendish Grande Naine (CAV; AAA) during early compatible interactions with M. incognita . Roots were analysed by brightfield light microscopy over a 35 d period to examine nematode penetration and morphological cell transformation. RNA samples were extracted 3, 7 and 10 days after inoculation (DAI) with nematode J2 juveniles, and cDNA libraries were sequenced using lllumina HiSeq technology. Sequences were mapped to the M. acuminata ssp. malaccensis var. Pahang genome sequence, differentially expressed genes (DEGs) identified and transcript representation determined by gene set enrichment and pathway mapping. Microscopic analysis revealed a life cycle of M. incognita completing in 24 d in CAV and 27 d in 4279-06. Comparable numbers of DEGs were up- and downregulated in each genotype, with potential involvement of many in early host defence responses involving reactive oxygen species and jasmonate/ethylene signalling. DEGs revealed concomitant auxin metabolism and cell wall modification processes likely to be involved in giant cell formation. Notable transcripts related to host defence included those coding for leucine-rich repeat receptor-like serine/threonine-protein kinases, peroxidases, thaumatin-like pathogenesis-related proteins, and DREB, ERF, MYB, NAC and WRKY transcription factors. Transcripts related to giant cell development included indole acetic acid-amido synthetase GH3.8 genes, involved in auxin metabolism, as well as genes encoding expansins and hydrolases, involved in cell wall modification. Expression analysis in M. acuminata during compatible interactions with RKNs provides insights into genes modulated during infection and giant cell formation. Increased understanding of both defence responses to limit parasitism during compatible interactions and effector-targeted host genes in this complex interaction will facilitate the development of genetic improvement measures for RKNs.

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.

Modifications were made in the PCR conditions aiming to overcome the problem of non-amplification of the Xanthomonas phaseoli pv. manihotis (Xpm) fragment, using the primer pair XV / XK described in the literature. The objective of this study was to propose changes in the primers already described (XV / XK_MOD) and validate the use of these new primers in identifying Xpm. The validation procedure was carried out with the primer pair XV and XK_MOD, using different strains of Xpm, other plant pathogenic and endophytic bacteria genera and one isolate of X. phaseoli pv. passiflorae. As a control, additional reactions were conducted in multiplex with the universal primers for the 16S rRNA gene of the bacteria together with XV / XK and XV / XK_MOD. Using the forward primer (XV) described in the literature together with the modified reverse primer (XK_MOD), it was possible to achieve amplification from DNA extracted from in vitro cultures and from infected tissue, but no amplification was noticed for the primer pair described in the literature, confirming the effectiveness of the proposed modification.

The cultivation of bananas and other plants is limited by environmental stresses caused by climate change. In order to recognize physiological, biochemical and molecular components indicated to confer tolerance to water stress in Musa spp. we present the first systematic review on the topic. A systematic literature review was conducted using four databases for academic research (Google Academic, Springer, CAPES Journal Portal and PubMed Central). In order to avoid publication bias, a previously established protocol and inclusion and exclusion criteria were used. The drought tolerance response is genotype-dependent, therefore the most studied varieties are constituted by the \"B\" genome. Tolerant plants are capable of super-expressing genes related to reisistance and defense response, maintaining the osmotic equilibrium and elimination of free radicals. Furthermore, they have higher amounts of water content, chlorophyll levels, stomatic conductance and dry root matter, when compared to susceptible plants. In recent years, few integrated studies on the effects of water stress on bananas have been carried out and none related to flood stress. Therefore, we highlight the need for new studies on the mechanisms of differentially expressed proteins in response to stress regulation, post-translational mechanisms and epigenetic inheritance in bananas.

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.

The present study evaluated the physiological, molecular and hormonal parameters from scion/rootstock interaction of citrus plants during recurrent water deficit. Responses of the Valencia (VO) scion variety grafted on two rootstocks with different soil water extraction capacities, Rangpur Lime (RL) and Sunki Maravilha (SM), during three successive periods of water deficit: plants exposed to a single episode of water deficit (WD1) and plants exposed to two (WD2) and three (WD3) recurrent periods of WD were compared. The combinations VO/RL and VO/SM presented polymorphic alterations of epigenetic marks and hormonal ( i . e . abscisic acid, auxins and salicylicacid) profiles, which were particularly prominent when VO/SM plantswere exposed toWD3 treatment. Upon successive drought events, the VO/SM combination presented acclimatization characteristics that enable higher tolerance to water deficit by increasing transpiration ( E ), stomatal conductance ( g s ) and photosynthetic rate ( A ), which in turn may have facilitated the whole plant survival. Besides providing comprehensive data on the scion/rootstock interactions upon successive stress events, this study brings the first dataset suggesting that epigenetic alterations in citrus plants triggered by recurrent water deficit lead to improved drought tolerance in this crop species.

Bananas are tropical fruits grown worldwide playing a key role in market trade and especially used as main food source for low income populations. In Brazil, bananas are mainly consumed in natura, occupying the second largest internal market. Nevertheless, this crop presents low availability of productive commercial varieties with good agronomic characteristics. A strategy undertaken to solve this problem is the development of new cultivars through conventional genetic breeding methods. However, this strategy presents some obstacles such as female sterility and low number of seeds. In order to overcome these shortcomings, use of mutation induction aiming the selection of mutants with desirable agronomic characteristics seems to have great potential for developing new cultivars. The objective of the present work was to evaluate the genetic variability in putative banana ‘Pacovan' (AAB genome, subgroup Prata Type) mutants submitted to gamma ray irradiation, using a set of agronomical and molecular data (ISSR markers). The distance between the putative ‘Pacovan' mutants varied from 0.26 to 0.64 with cophenetic correlation coefficient of 0.7669. Four mutants were selected based on best agronomical characteristics and height. This data also shows that there is variability that can be explored after the irradiation of ‘Pacovan' banana mutants, which can be used in the genetic breeding program of banana aiming to develop short new varieties that also present good agronomic characteristics. This is the first attempt to use combined data in order to evaluate the genetic variability in putative banana mutants.

Cassava root-rot incited by soil-borne pathogens is one of the major diseases that reduces root yield. Although the use of resistant cultivars is the most effective method of management, the genetic basis for root-rot resistance remains poorly understood. Therefore, our work analyzed the transcriptome of two contrasting genotypes (BRS Kiriris/resistant and BGM-1345/susceptible) using RNA-Seq to understand the molecular response and identify candidate genes for resistance. Cassava seedlings (resistant and susceptible to root-rot) were both planted in infested and sterilized soil and samples from Initial-time and Final-time periods, pooled. Two controls were used: (i) seedlings collected before planting in infested soil (absolute control) and, (ii) plants grown in sterilized soil (mock treatments). For the differentially expressed genes (DEGs) analysis 23.912 were expressed in the resistant genotype, where 10.307 were differentially expressed in the control treatment, 15 DEGs in the Initial Time-period and 366 DEGs in the Final Time-period. Eighteen candidate genes from the resistant genotype were related to plant defense, such as the MLP-like protein 31 and the peroxidase A2-like gene. This is the first model of resistance at the transcriptional level proposed for the cassava × root-rot pathosystem. Gene validation will contribute to screening for resistance of germplasm, segregating populations and/or use in gene editing in the pursuit to develop most promising cassava clones with resistance to root-rot.

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.