This study focused on the in vitro propagation and DNA barcoding of the endangered herb Solanum corymbosum from the La Joya Desert, Peru. The primary objective of this study was to establish a protocol for the in vitro conservation and molecular identification of these species. This study used cytokinins, specifically BAP and TDZ, for shoot regeneration from leaf explants, achieving high rates of shoot regeneration and proliferation. The optimal concentration of TDZ (1 mg/L) resulted in 88.7% regeneration. In the rooting phase, auxins, specifically, IBA and NAA, were used, achieving high rooting rates and root numbers. The optimal concentration of IBA (2 mg/L) resulted in 100% rooting and 9.8 roots per shoot. DNA barcoding with four cpDNA markers (matK, rbcL, trnL-trnF, and psbA-trnH) allowed accurate molecular identification, confirming species identity within the genus Solanum. Key results included successful in vitro propagation with high rates of shoot formation and rooting and 100% survival during acclimatization. Molecular analysis confirmed the taxonomic identification of S. corymbosum, with high percentages of similarity (97% to 100%) with other Solanum species. Phylogenetic analysis confirmed its classification within the Radicans clade, sharing its characteristics with other Andean species of the genus Solanum. This study revealed that the propagation in vitro protocol and DNA barcoding are effective tools for the conservation of S. corymbosum, offering valuable methods for the conservation and genetic study of this and other threatened plant species.

Rapid alkalinization factors (RALFs) are ubiquitous cysteine-rich peptides present in plants. They exert diverse functions as hormonal signals in various processes, including cell growth, root elongation, and fertilization. RALF peptides can also act as negative regulators of the plant immune response, inhibiting the formation of the signal receptor complex for immune activation. In Fragaria × ananassa, silencing of FaRALF33 gene plays a key role in the defense against the fungal pathogen Colletotrichum acutatum. In this study, single-guide RNA (sgRNAs) were designed in silico for clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) 9-mediated FaRALF33 gene mutagenesis in F. × ananassa for the reduction of C. acutatum infection. FaRALF33 was compared with homologous RALF33 sequences from other plant species, showing that the amino acid sequence of FaRALF33 presents typical sequences of known RALF peptides in RRILA proteolytic site, in addition to tight clustering presented by FaRALF33 with FvRALF33. The online tool CHOPCHOP provided 73 hits for FaRALF33 gene, selecting two sgRNA sequences for mutagenesis, sgRNA 1 (5’-CGACTCTCCCATCTCTTGGACT-3’) and sgRNA 2 (5’-GCAAGCAACGGCAGCGATCA-3’). The predicted secondary structures of the selected sgRNAs presented efficient structures in targeted mutagenesis. The pCas9-TPC-GFP-2XsgRNA vector for CRISPR/Cas9-mediated FaRALF33 gene mutagenesis was designed in silico with two sgRNA sequences (with Arabidopsis thaliana U6-26 promoters) and a green fluorescent protein marker.

Sacha inchi (Plukenetia volubilis) is a high-value crop due to its high content of omega-3 fatty acids and its outstanding nutritional, pharmaceutical, and cosmetic properties. However, this species faces challenges from diseases, particularly root rot. In this study, we identified one of the causal agents of root rot in sacha inchi using morphological observations, molecular methods, and pathogenicity tests. The pathogen was isolated from root tissues showing symptoms of Fusarium infection, observed in a plot in the Picota province. Morphological identification, DNA sequencing, and phylogenetic analysis using the ITS and TEF-1α markers revealed that the isolate causing root rot was Fusarium suttonianum (FSSC 20). Analysis of the PQ636870 (ITS) and PQ639345 (TEF-1α) sequences in the NCBI database, together with phylogenetic analysis, revealed 99.58% and 99.51% similarity with the ITS and TEF sequences, respectively, corresponding to F. suttonianum. Pathogenicity tests confirmed that this species induced the same symptoms observed in the field, fulfilling Koch’s postulates. This study represents the first report of F. suttonianum as a pathogen causing root rot in sacha inchi in Peru. This finding is critical for developing effective strategies for disease management and control, contributing to the sustainability and improvement of sacha inchi production in the region.

The wild dragon fruit (Selenicereus spp.) with orange peel and fuchsia pulp, known as “Naranja de Churuja”, represents a promising plant genetic resource from the Amazonas region of Peru. The objective of this study was to develop an in vitro micropropagation protocol for this local accession by evaluating shoot induction and proliferation, in vitro rooting, and ex vitro acclimatization. During the multiplication phase, nine combinations of 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA) were evaluated using cladode segments. Shoot induction was high in most treatments (95.56–100.00%), and the combination of 2.0 mg L⁻¹ BAP + 0.1 mg L⁻¹ IBA produced the highest number of shoots per explant (9.67), whereas 1.0 mg L⁻¹ BAP without IBA promoted the greatest shoot elongation (2.34 cm). During the rooting phase, 1.50 mg L⁻¹ IBA produced the highest number of roots per explant (3.41), whereas 1.00 mg L⁻¹ IBA generated the greatest shoot length (4.12 cm). During ex vitro acclimatization, all plants survived (100% survival), but growth depended significantly on the substrate, with the moss treatment promoting the greatest development. In conclusion, an efficient micropropagation protocol was established for the wild “Naranja de Churuja” accession, with potential for mass propagation, germplasm conservation, and future biotechnological applications, including genetic transformation and genome editing.