Dwinita Wikan Utami, Adhityo Wicaksono, M. Khais Prayoga, Heri Syahrian, Vitria P. Rahadi, Erdiansyah Rezamela, Budi Martono, Tri Joko Santoso, Nur Kholilatul Izzah, Haris Maulana, Ady Daryanto, Rerenstradika Tizar Terryana, Imas Rita Saadah, David Virya Chen

<br />High-throughput molecular technology DArTseq generates markers for potential use in molecular breeding of crops. Using DArTseq, we analysed a comprehensive set of 333 European potato varieties reflecting the outcomes of long-term breeding history and representing a potential germplasm for future breeding of potatoes in the Central European region. The varieties were classified according to four factors: region of origin, breeder, earliness and utilisation mode, that may potentially reflect their genetic structure, and for which complete data were publicly available. The DArTseq analysis was performed by the service centre, the Diversity Array Technology (University of Canberra), which generated approximately 38 000 silicoDArT and 64 000 SNP (single nucleotide polymorphism) polymorphic markers. The discriminatory ability of the markers in relation to the factors was confirmed using neighbour-joining and principal coordinate analysis (PCoA), while the informativeness was assessed using the discriminant analysis of principal components (DAPC). The analyses identified the 50 SNPs most strongly associated with each factor, along with their highly probable chromosomal localisation. Herein presented research contributes to the evaluation of potato genetic resources by adding the novel molecular data of active germplasm and implies their future utilisation in genome wide association studies and marker assisted selection.

The genus <i>Solanum</i> comprises numerous wild and cultivated species that are important for potato breeding. This pilot-scale study aimed to evaluate the genetic diversity in 44 accessions from <i>Solanum</i> sect. Petota, comprising wild species, Andean landraces, and modern cultivars, obtained from the Potato Research Institute Havlkv Brod, Ltd. and the Department of Crop Sciences and Agroforestry at the Faculty of Tropical AgriSciences, Czech University of Life Sciences Prague. Nuclear microsatellite markers (SSR, 29 loci) were applied via five multiplex PCR reactions and analysed using capillary electrophoresis. Binary data matrices were analysed using DARwin software to generate dendrograms reflecting allelic polymorphism. The SSR panel effectively differentiated cultivated accessions from wild types, consistent with the current taxonomy of the genus <i>Solanum</i>, with particularly clear clustering of Andean landraces and modern varieties. However, resolution among wild accessions was limited, likely due to their high genetic complexity and interspecific overlap. These results support the suitability of the SSR panel for analysing diversity in cultivated potatoes. while also highlighting the challenges in resolving wild <i>Solanum</i> taxa. This study contributes to germplasm characterisation and provides a molecular basis for future breeding programmes.

Efi Sarri, Anastasios Katsileros, Sofia Migardou, Panagiotis Viliotis, Ioannis Sidiropoulos, Dimitris Sifnaios, Pavlos Diamantis, Nikolaos Sklavounos, Eleni M. Abraham, Penelope J. Bebeli, Nasya Tomlekova, Dimosthenis Kizis, Eleni Tani

Rapeseed (<i>Brassica napus</i> L.) isamajor global oilseed crop and exhibits significant heterosis. The discovery and characterisation ofnovel male-sterile accessions remain fundamental for harnessing heterosis inrapeseed breeding. Previously, wedeveloped amale sterile two-type line system 19F08AB in<i>B. napus</i>. Inthis study, anther abortion in19F08A was characterised using the squash method. The inheritance ofmale sterility in19F08A and its genetic relationship toreported male sterile accessions inrapeseed was investigated using classical genetic analysis and male-sterility-gene-specific molecular markers. Results indicated that male sterile flowers of19F08A exhibit flat petals, reduced floral organs, short filaments, and completely degenerated stamens devoid ofpollen. Pollen mother cells in19F08A degenerated atthe pre-meiotic stage and aborted completely atthe tetrad stage, with nodyad ortetrad formation observed. This suggested that 19F08A represents ameiosis abnormality-type male sterility. Classical genetic and molecular marker analysis revealed that male-sterile plants 19F08A carry the genotype of<i>pol</i> (<i>RfpRfpMsms</i>), whereas fertile plants 19F08B possess <i>pol</i> (<i>RfpRfpmsms</i>). The effect ofthe <i>pol</i> cytoplasm was masked bythe <i>Rfp</i> gene. Therefore, fertility in19F08AB iscontrolled byapair ofnuclear genes (<i>Ms</i>/<i>ms</i>), with male sterility exhibiting dominance over fertility. The application prospects ofthis male-sterile accession are also discussed. These findings expand the pool ofmale-sterile resources available for <i>B. napus</i> hybrid breeding and contribute to plant male sterility theory.

<br />The present study investigates the differential protein expression levels between the frost-tolerant variety Qingda No.1 and the frost-sensitive variety Gannong No.9. The analysis was conducted using 4D-Label-free quantitative proteomics technology, with the samples collected prior to and after overwintering. The results showed that the protein expression of Qingda No.1 changed more significantly during the overwintering process, with 451 differentially expressed proteins (DEPs) being identified, of which 224 were up-regulated and 227 down-regulated. In contrast, the protein expression of Gannong No.9 differed from that of alfalfa roots of the frost-sensitive variety, with 204 DEPs being identified, of which 93 were up-regulated and 111 down-regulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the phenylpropanoid biosynthesis pathway was significantly enriched in both groups. Furthermore, enzymes such as phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase (C4H), and chalcone isomerase (CHI) were significantly up-regulated in the freezing-tolerant varieties. Protein interaction network analysis revealed the synergistic regulation mode of DEPs. The present study elucidated the metabolic adaptive mechanism of overwintering in alfalfa roots at the protein level, thus providing a theoretical basis for the selection and breeding of frost-tolerant varieties.

This study has evaluated the genetic characteristics and wheat processing-related properties of four Korean wheat landraces (KWLs). The KWLs were found to possess the vernalization alleles vrn-A1, vrn-B1, and Vrn-D1 and the photoperiod alleles Ppd-A1b, Ppd-B1b, and Ppd-D1a. The Korean cultivated variety Keumgang also shared these alleles with the exception of vrn-D1. With regard to grain hardness, KWL 2 was shown to possess Pina-D1a and Pinb-D1b like Keumgang, while other KWLs were classified as carrying Pina-D1a and Pinb-D1a. All KWLs were found to be non-waxy, carrying the alleles Wx-A1a, Wx-B1a, and Wx-D1a. With regard to the polyphenol oxidase (PPO) genes, all four KWLs carried low-activity alleles, in contrast to the Keumgang sample. The assessment of physicochemical properties revealed that KWL 1, 3, and 4 had a higher amylose content but a lower protein content than KWL 2 and Keumgang. In tests of solvent retention capacity KWL 1 and KWL 2 exhibited the lowest and highest values, respectively, for all four solvents used in the tests. With regard to the dough properties, the results of Mixolab analysis indicated a faster starch gelatinisation in KWL1, while in KWL 2 a high water absorption and the longest dough development and stability times were found. KWL 3 and 4 exhibited similar dough behaviours. Principal component analysis of the four KWL lines revealed distinct clustering based on their physicochemical and dough-related traits.

Foxtail millet (<i>Setaria italica</i>) isaresilient yet underutilised C₄cereal valued for its adaptability toabiotic stress and high nutritional content. While panicle traits have been linked toyield inmany cereals, the spatial arrangement offoxtail millet panicles remained unexplored, especially among locally adapted genotypes. This study aimed tocharacterise spatial panicle architecture traits and toanalyse the <i>DUF-640</i>, agene controlling primary branch number, among Indonesian foxtail millet genotypes. Results revealed substantial variation inpanicle architecture, including primary branch number, grain number, and grain density ineight Indonesian foxtail millet genotypes, suggesting potentially greater diversity across broader germplasm. Incontrast tothe substantial panicle trait variations, phylogenetic and structural analyses showed that <i>DUF-640</i> genes were highly conserved across <i>Setaria</i> species. Although previously associated with primary branch development, the coding sequence of<i>DUF-640</i> was not associated with branching variation inthis study. However, its high sequence conservation across <i>Setaria</i> species suggests avital and possibly conserved regulatory function. This study enhances the understanding ofthe morphological and genetic diversity offoxtail millet, particularly among Indonesian foxtail millet genotypes. Future research should focus onthe functional characterisation of<i>DUF-640</i> and the identification of regulatory sequences governing its gene expression.