Newly Developed Restorer Lines of Sorghum [Sorghum bicolor (L.) Moench] Resistant to Greenbug.

Eight lines of grain sorghum [Sorghum bicolor (L.) Moench], which can be used as a promising source material in heterotic hybrid breeding as pollen fertility restorers and donors of resistance to the greenbug (Schizaphis graminum Rondani), are characterized. The new restorer lines (R-lines) were developed by crossing the maternal sterile line Nizkorosloe 81s (CMS A1) with two lines selected from the grain sorghum collection accessions VIR-928 and VIR-929 as the paternal forms. The R-lines were genotyped using PCR markers, and also characterized by height, duration of the seedling-flowering period, and some of the technological properties of flour. With the use of microsatellite markers linked to the Rf genes and by hybridological analysis, it was shown that the new lines carry the dominant allele of the gene Rf2. The PCoA analysis demonstrated clear differences of each R-line from the parents. The genotypes of the new lines and their parental forms for the Rf2 locus were confirmed by applying three allele-specific codominant CAPS markers which detected SNPs in the candidate Rf2 gene. All new lines were highly fertile, as demonstrated by cytological analysis of acetocarmine-stained pollen preparations. A high resistance to the greenbug was demonstrated for each new R-line both in the laboratory and field conditions against a severe aphid infestation. Grain quality parameters such as protein content and dough rheological properties varied widely and were quite satisfactory in some R-lines. Characteristics common to all eight sorghum lines studied, such as the ability to restore pollen fertility in the F1 generation, good pollen quality, greenbug resistance, early ripening, spreading panicle, and low stature, allow us to recommend them for producing commercial F1 hybrids with satisfactory grain quality for both fodder and food purposes.

Genetic Diversity of Barley Accessions from East Asia for Greenbug Resistance.

The greenbug, Schizaphis graminum, is a dangerous pest of barley and other grain crops in the south of Russia. An effective and environmentally friendly way to control this insect is to cultivate resistant varieties. The differential interaction between the phytophage and host plants necessitates the search for new donors of resistance. Seven hundred and seventy-eight accessions of barley from East Asian countries (313 from China, 450 from Japan, and 15 from Nepal) were evaluated for greenbug resistance. The Krasnodar population of the insect and clones isolated from it were used in the experiments. Forty heterogeneous accessions were identified, in which plants with a high level of resistance to the aphid were found. As a result of damage assessment by the 108 S. graminum clones of 11 lines selected from heterogeneous accessions, 52 insect virulence phenotypes were identified. Experiments with aphid test clones showed that all 11 lines possess diverse greenbug resistance alleles, which differ from the previously identified Rsg1, but their efficiency is low. The frequency of clones virulent to ten lines and the cultivar Post (a carrier of the Rsg1 gene) varies from 60.4% to 98.0%. The exception is line 15903, which is resistant to the aphid population and protected by one dominant gene. The high resistance of other lines against a part of the natural population of S. graminum is also under oligogenic control. Lines 15600 and 16190 each have one dominant resistance gene, and line 28129 is protected by two genes, the dominant and recessive ones. A recessive resistance gene is presumably present in line 15600. Lines 16237/1 and 16237/2, isolated from the same collection accession, each have one dominant gene effective against individual aphid clones. The loss of effectiveness of distinctly manifested resistance genes causes the expression of previously masked genes with a weak phenotypic manifestation, which differentially interact with insect genotypes.

Refinement of Rf1-gene localization and development of the new molecular markers for fertility restoration in sunflower.

BACKGROUND: Ability to restore male fertility is important trait for sunflower breeding. The most commonly used fertility restoration gene in the production of sunflower hybrids is Rf1. The localization of Rf1 on the linkage group 13 has been previously shown, however, its exact position, its sequence and molecular mechanism for fertility restoration remain unknown. Therefore, several markers linked to Rf1 gene, commonly used for MAS, don't always allow to identify the genotype of plants. For this reason, the search for new markers and precise localization of the Rf1 gene is an urgent task. METHODS AND RESULTS: Based on previously identified single nucleotide polymorphisms (SNPs) at LG13, significantly associated with the ability to restore male fertility, two markers have been developed that have performed well after careful evaluation. These markers, together with other Rf1 markers, were applied for genotyping 72 diversity panel accessions and 291 individuals of F2 segregating population, obtained from crossing the cytoplasmic male sterility (CMS) AHO33 and restorer RT085HO lines. The analysis revealed no recombinants between Rf1 gene and SRF833 marker, the distance between Rf1 and SRF122 marker was 1.0 cM. CONCLUSIONS: Data obtained made it possible to specify the localization of the Rf1 gene and reduce the list of candidate genes to the 3 closely linked PPR-genes spanning a total of 59 Kb. However, it cannot be ruled out that analysis of the candidate region in the genome of fertility restorer lines can reveal new candidate genes in this locus that are absent in the cytoplasmic male sterility maintainer reference sequence.

Genetic Resources of Cereal Crops for Aphid Resistance.

The genetic resources of cereal crops in terms of resistance to aphids are reviewed. Phytosanitary destabilization led to a significant increase in the harmfulness of this group of insects. The breeding of resistant plant genotypes is a radical, the cheapest, and environmentally safe way of pest control. The genetic homogeneity of crops hastens the adaptive microevolution of harmful organisms. Both major and minor aphid resistance genes of cereal plants interact with insects differentially. Therefore, rational breeding envisages the expansion of the genetic diversity of cultivated varieties. The possibilities of replenishing the stock of effective resistance genes by studying the collection of cultivated cereals, introgression, and creating mutant forms are considered. The interaction of insects with plants is subject to the gene-for-gene relationship. Plant resistance genes are characterized by close linkage and multiple allelism. The realizing plant genotype depends on the phytophage biotype. Information about the mechanisms of constitutional and induced plant resistance is discussed. Resistance genes differ in terms of stability of expression. The duration of the period when varieties remain resistant is not related either to its phenotypic manifestation or to the number of resistance genes. One explanation for the phenomenon of durable resistance is the association of the virulence mutation with pest viability.

Genotyping and lipid profiling of 601 cultivated sunflower lines reveals novel genetic determinants of oil fatty acid content.

BACKGROUND: Sunflower is an important oilseed crop domesticated in North America approximately 4000 years ago. During the last century, oil content in sunflower was under strong selection. Further improvement of oil properties achieved by modulating its fatty acid composition is one of the main directions in modern oilseed crop breeding. RESULTS: We searched for the genetic basis of fatty acid content variation by genotyping 601 inbred sunflower lines and assessing their lipid and fatty acid composition. Our genome-wide association analysis based on the genotypes for 15,483 SNPs and the concentrations of 23 fatty acids, including minor fatty acids, revealed significant genetic associations for eleven of them. Identified genomic regions included the loci involved in rare fatty acids variation on chromosomes 3 and 14, explaining up to 34.5% of the total variation of docosanoic acid (22:0) in sunflower oil. CONCLUSIONS: This is the first large scale implementation of high-throughput lipidomic profiling to sunflower germplasm characterization. This study contributes to the genetic characterization of Russian sunflower collections, which made a substantial contribution to the development of sunflower as the oilseed crop worldwide, and provides new insights into the genetic control of oil composition that can be implemented in future studies.

Nucleotide sequence polymorphism in the RFL-PPR genes of potato.

Cytoplasmic male sterility (CMS) is widely used for hybrid seed production in cultivated Solanaceae species. However, there is very limited information about CMS-Rf genetic systems in potato (Solanum tuberosum). Studying the CMS-Rf systems in potato is both of theoretical and practical significance due to the emergence of a new revolutionary strategy of reinventing potato as adiploid inbred line-based crop to develop F1 hybrid seed potato breeding (Lindhout et al. 2011; Jansky et al. 2016). To search for potato Rf gene candidates, the comparative genetic approach was applied. Based on similarity to petunia Rf-PPR592 gene, 38 fragments were identified in five loci of the whole-genome nucleotide sequence of the accession DM 1-3 516 R44 S. tuberosum Phureja group (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The putative encoded mitochondrial proteins have 589-597 amino acid residues, similarto RF-PPR proteins of petunia and chili pepper and contain 14 or 15 PPR motifs. Primers have been developed flanking the most variable 782-865 bp regions of the selected loci, and polymorphism of the cloned fragments has been investigated in a subset of nine potato genotypes. The amplified fragments included seven or eight PPR motifs and lacked introns. The SNP frequencies ranged from 7.0 to 19.8% depending on the locus, while the ratio of nonsynonymous to synonymous substitutions varied between 0.9 and 2.1. Positions 1, 3 and 6 were the most variable in the studied PPR motifs. Our results demonstrated that the analysed sequences belong to the RFL-PPR gene subfamily and may be considered as Rf gene candidates in potato.

Serine proteinase inhibitors in the Compositae: distribution, polymorphism and properties.

Multiple molecular forms of inhibitors of trypsin (TI) and chymotrypsin (CI), which are typical digestive enzymes of insects, mammals and micro-organisms, and subtilisin (SI), a proteinase of many bacteria and phytopathogenic fungi, were identified in seeds and vegetative organs of the majority of 128 wild and cultivated species representing 65 genera of three of the subfamilies of the Compositae. Inhibitors with M(r) ranging from 7450 to 7800 and combining activities towards subtilisin and trypsin and/or chymotrypsin (T/C/SI) had the widest distribution and may be involved in plant defense mechanisms. They were found in many species of the subfamilies Carduoideae (genera Carthamus, Centaurea, Cirsium), Cichorioideae (Lactuca, Taraxacum) and Asteroideae (Helianthus, Cosmos, Bidens). Partial amino acid sequencing showed that the safflower (Carthamus tinctorius) T/C/SI and Cosmos bipinnatus T/C/SI, T/SI and C/SI belonged to the potato I inhibitor family. The most active, variable and heterogeneous inhibitors were found in species of the tribe Heliantheae, which is placed in the evolutionary advanced subfamily Asteroideae. Seeds of Helianthus species, Eclipta prostrata, Gailardia aristata, Zinnia elegans and Silphium perfoliatum contained various TI with M(r) ranging from 1500 to 14,750, with some also containing SI. H. annuus seeds contain a unique cyclic TI of M(r) 1514 and similar TI were also present in other Helianthus spp. and the related species Tithonia diversifolia. Zinnia elegans contained a TI with M(r) 11,350 which appeared to represent a novel type of inhibitor distantly related to the cereal subgroup of Bowman-Birk inhibitors. TI and T/SI varied widely in H. annuus lines and wild Helianthus species in their presence or absence and composition. Similar T/SI components were found in the cultivated diploid H. annuus and annual diploid species with the B genome but not in perennials with the A genome. Some T/SI, SI and TI were detected in vegetative organs of sunflower and other Compositae. Studies of the polymorphism and distribution of proteinase inhibitors are relevant to the evolution of protective protein systems and the mechanisms of resistance to pathogenic organisms in the Compositae and other plants.