Major chromosome rearrangements in intergeneric wheat × rye hybrids in compatible and incompatible crosses detected by GBS read coverage analysis.

The presence of incompatibility alleles in primary amphidiploids constitutes a reproductive barrier in newly synthesized wheat-rye hybrids. To overcome this barrier, the genome stabilization process includes large-scale chromosome rearrangements. In incompatible crosses resulting in fertile amphidiploids, the elimination of one of the incompatible alleles Eml-A1 or Eml-R1b can occur already in the somatic tissue of the wheat × rye hybrid embryo. We observed that the interaction of incompatible loci Eml-A1 of wheat and Eml-R1b of rye after overcoming embryo lethality leads to hybrid sterility in primary triticale. During subsequent seed reproductions (R1, R2 or R3) most of the chromosomes of A, B, D and R subgenomes undergo rearrangement or eliminations to increase the fertility of the amphidiploid by natural selection. Genotyping-by-sequencing (GBS) coverage analysis showed that improved fertility is associated with the elimination of entire and partial chromosomes carrying factors that either cause the disruption of plant development in hybrid plants or lead to the restoration of the euploid number of chromosomes (2n = 56) in the absence of one of the incompatible alleles. Highly fertile offspring obtained in compatible and incompatible crosses can be successfully adapted for the production of triticale pre-breeding stocks.

Genetic enhancement of okra [Abelmoschus esculentus (L.) Moench] germplasm through wide hybridization.

Introduction: The introgression of genetic material from one species to another through wide hybridization and repeated back-crossing, plays an important role in genetic modification and enriching the cultivated gene-pool with novel genetic variations. Okra (Abelmoschus esculentus [(L.) Moench)] is a popular vegetable crop with high dietary fibre and protein, rich in essential amino acids, lysine and tryptophan. The wild Abelmoschus genepool has many desirable traits like ornamental value, short internodal length, more number of productive branches, extended bearing, perennation tendency, reduced fruit length (more consumer preferred trait), high mucilage content (medicinal value), abiotic stress tolerances such as drought, high temperature and biotic stress resistances such as okra Yellow Vein Mosaic Virus (YVMV) and Enation Leaf Curl Virus (ELCV) diseases. The repeated use of elite breeding lines led to narrowing of the genetic base of the okra crop, one of the major factors attributed to breakdown of resistance/ tolerance to biotic stresses. YVMV and ELCV are the two major diseases, causing significant yield loss in okra. Hence, wide hybridization was attempted to transfer tolerance genes from wild species to the cultivated genepool to widen the genetic base. Material and methods: The screening of germplasm of wild Abelmoschus species at hotspots led to the identification of tolerant species (Abelmoschus pungens var. mizoramensis, A. enbeepeegeearensis, A. caillei, A. tetraphyllus and A. angulosus var. grandiflorus), which were further used in a wide-hybridization programme to generate interspecific hybrids with the cultivated okra. Presence of pre- and post-zygotic barriers to interspecific geneflow, differences in ploidy levels and genotype specific variations in chromosome numbers led to varying degrees of sterility in F1 plants of interspecific crosses. This was overcome by doubling the chromosome number of interspecific hybrids by applying Colchicine at the seedling stage. The 113 cross derivatives generated comprising amphidiploids in the F1 generation (30), F3 (14), one each in F2 and F4 generations, back cross generation in BC1F2 (03), BC1F3 (25), and BC2F3 (02), crosses between amphidiploids (27), multi-cross combinations (07) and inter-specific cross (between A. sagittifolius × A. moschatus subsp. moschatus) selfed derivatives at F8 generation (03) were characterized in the present study. Besides they were advanced through selfing and backcrossing. Results and Discussion: The amphidiploids were found to possess many desirable genes with a considerable magnitude of linkage drag. Majority of the wide cross derivatives had an intermediate fruit morphology and dominance of wild characters viz., hispid fruits, stem, leaves, tough fruit fibre, vigorous perennial growth habit and prolonged flowering and fruiting. The fruit morphology of three BC progenies exhibited a high morphological resemblance to the cultivated okra, confirming successful transfer of useful genes to the cultivated okra genepool. The detailed morphological characteristics of the various combinations of Abelmoschus amphidiploids and the genetic enhancement of the genepool achieved in this process is reported here.

Altered Developmental and Metabolic Gene Expression in Basil Interspecific Hybrids.

To understand the altered developmental changes and associated gene expression in inter-genomic combinations, a study was planned in two diverse yet closely related species of Ocimum, targeting their hybrid F1 and amphidiploids. The existing developmental variations between F1 and amphidiploids was analyzed through phenotypical and anatomical assessments. The absence of 8330 transcripts of F1 in amphidiploids and the exclusive presence of two transcripts related to WNK lysine-deficient protein kinase and geranylgeranyl transferase type-2 subunit beta 1-like proteins in amphidiploids provided a set of genes to compare the suppressed and activated functions between F1 and amphidiploids. The estimation of eugenol and methyleugenol, flavonoid, lignin and chlorophyll content was correlated with the average FPKM and differential gene expression values and further validated through qRT-PCR. Differentially expressed genes of stomatal patterning and development explained the higher density of stomata in F1 and the larger size of stomata in amphidiploids. Gene expression study of several transcription factors putatively involved in the growth and developmental processes of plants clearly amalgamates the transcriptome data linking the phenotypic differences in F1 and amphidiploids. This investigation describes the influence of interspecific hybridization on genes and transcription factors leading to developmental changes and alleviation of intergenomic instability in amphidiploids.

Karyotype Reorganization in Wheat-Rye Hybrids Obtained via Unreduced Gametes: Is There a Limit to the Chromosome Number in Triticale?

To date, few data have been accumulated on the contribution of meiotic restitution to the formation of Triticum aestivum hybrid karyotypes. In this study, based on FISH and C-banding, karyotype reorganization was observed in three groups of F5 wheat-rye hybrids 1R(1A) × R. Aberrations, including aneuploidy, telocentrics, and Robertsonian translocations, were detected in all groups. Some of the Group 1 plants and all of the Group 2 plants only had a 4R4R pair (in addition to 1R1R), which was either added or substituted for its homeolog in ABD subgenomes. In about 82% of meiocytes, 4R4R formed bivalents, which indicates its competitiveness. The rest of the Group 1 plants had 2R and 7R chromosomes in addition to 1R1R. Group 3 retained all their rye chromosomes, with a small aneuploidy on the wheat chromosomes. A feature of the meiosis in the Group 3 plants was asynchronous cell division and omission of the second division. Diploid gametes did not form because of the significant disturbances during gametogenesis. As a result, the frequency of occurrence of the formed dyads was negatively correlated (r = -0.73) with the seed sets. Thus, meiotic restitution in the 8n triticale does not contribute to fertility or increased ploidy in subsequent generations.

Development and characterization of Triticum turgidum - Aegilops comosa and T. turgidum - Ae. markgrafii amphidiploids.

Aegilops comosa and Ae. markgrafii are diploid progenitors of polyploidy species of Aegilops sharing M and C genomes, respectively. Transferring valuable genes/traits from Aegilops into wheat is an alternative strategy for wheat genetic improvement. The amphidiploids between diploid species of Aegilops and tetraploid wheat can act as bridges to overcome obstacles from direct hybridization and can be developed by the union of unreduced gametes. In this study, we developed seven Triticum turgidum - Ae. comosa and two T. turgidum - Ae. markgrafii amphidiploids. The unreduced gametes mechanisms, including first-division restitution (FDR) and single-division meiosis (SDM), were observed in triploid F1 hybrids of T. turgidum - Ae. comosa (STM) and T. turgidum - Ae. markgrafii (STC). Only FDR was observed in STC hybrids, whereas FDR or both FDR and SDM were detected in the STM hybrids. All seven pairs of M chromosomes of Ae. comosa and C chromosomes of Ae. markgrafii were distinguished by fluorescent in situ hybridization (FISH) probes pSc119.2 and pTa71 combinations with pTa-535 and (CTT)12/(ACT)7, respectively. Meanwhile, the chromosomes of tetraploid wheat and diploid Aegilops parents were distinguished by the same FISH probes. The amphidiploids possessed specific valuable traits such as multiple tillers, large seed size related traits, and stripe rust resistance that could be utilized in the genetic improvement of wheat.

Beta-amylase gene variability in introgressive wheat lines.

Variability of the beta-amylase gene in bread wheat, artificial amphidiploids, and derived introgression wheat lines was analyzed. Variation in homeologous beta-amylase sequences caused by the presence of MITE (Miniature Inverted-Repeat Transposable Element) and its footprint has been identified in bread wheat. The previously unknown location of MITE in Triticum urartu and T. aestivum L. beta-amylase gene has been found. These species have a MITE sequence in the third intron of beta-amylase, as opposed to Aegilops comosa and a number of other Triticeae species, which have it in the fourth intron. These two MITEs from Ae. comosa and T. aestivum were shown to have low identity scores. Miosa, an artificial amphidiploid, which has the M genome from Ae. comosa was shown to lose the MITE sequences. This loss might be caused by genomic shock due to allopolyploidization.

Resistência à mancha preta e qualidade agronômica de plantas rc1f2 de cruzamentos do híbrido anfidiplóide (Arachis ipaensis x A. duranensis) com o amendoim cultivado (A. hypogaea) / Resistance to late leafspot and agronomic quality of bc1f2 plants from crosses between the amphyploids hibrid (arachis ipaensis x a. duranensis) and the cultivated peanut (a. hypogaea)

A resistência à mancha preta (Cercosporidium personatum) e a condição agronômica de plantas de amendoim foram avaliadas em populações RC1F2 obtidas de cruzamentos do anfidiplóide (Arachis ipaensis x Arachis duranensis) com o genitor recorrente, cultivar Runner IAC 886. O experimento foi conduzido em condições de campo, em Pindorama, SP., sob condições naturais de ocorrência da doença. A doença foi avaliada em cada planta, em duas épocas, usando uma escala de notas visuais de 1 a 9, considerando o número de lesões nas folhas e a desfolha causada pela doença. Foi realizada após a colheita, em 90 plantas tomadas ao acaso, a avaliação da produção de vagens e grãos/planta, o rendimento de grãos após descascamento, massa média de grãos, número de vagens/planta, número e porcentagem de vagens com dois grãos. Os resultados mostraram ampla variabilidade para resistência à doença. Dependendo do genitor hypogaea utilizado nos cruzamentos iniciais com o anfidiplóide, 4,2 a 7,7% de plantas portadoras de resistência próxima a do parental silvestre resistente (A. ipaensis) foram resgatadas em F2. Observaram-se diferenças significativas entre plantas RC1F2 em função da planta F1 da qual descendem, tanto para resistência à doença como para caracteres agronômicos, tendo em vista a condição segregante das plantas F4 utilizadas nos cruzamentos com o genitor recorrente, cultivar Runner IAC 886. As correlações negativas observadas entre resistência à doença e caracteres agronômicos como produção de vagens e grãos/planta e massa média de grãos sugerem a necessidade de novo ciclo de retrocruzamentos para quebrar eventuais ligações indesejáveis e gerar indivíduos resistentes mais próximos do padrão comercial.

Resistance to late leafspot (Cercosporidium personatum) and agronomic condition of peanut plants were assessed in segregating RC1F2 populations obtained from crosses between the amphidiploid (A. ipaensis x A. duranensis) and the recurrent parent, cultivar Runner IAC 886. The experiment was carried out at Pindorama, SP, under natural field conditions of the disease. Disease was evaluated at two dates during the growing cycle, in 1,500 plants, using a scale of visual notes ranging from 1 to 9, and taking into consideration the number of lesions in leaves and the degree of defoliation caused by the disease. The data obtained were used to estimate the AUDPC (Area Under the Disease Progress Curve). Agronomic evaluations were performed after harvesting, in 90 randomly taken plants. The following agronomic characters were evaluated: pod and kernel production per plant, shelling percentage, number of pods per plant, number and percentage of two-seed pods. The results showed wide variability for resistance to the disease. Depending on the hypogaea parent used in the initial crosses with the amphidiploids, 4,2 to 7,7% of plants with disease resistance close to the wild resistant parent (A. ipaensis) could be rescued in the F2 generation. Significant differences were observed between RC1F2 plants descendent from the same F1 plant, in either disease resistance or in agronomic traits, due to the segregant condition of the F4 plants used in the crosses to the recurrent parent, cultivar Runner IAC 886. Some of the RC1F1:2 plant families exhibited agronomic condition close to the cultivar IAC Caiapó, especially in production of pods and kernels, shelling percentage and average kernel weight. However, the large majority of the segregants in this generation still showed high percentages of one-seed pods. This condition, and the negative correlations observed between resistance to the disease and agronomic traits such as pod and kernel production per plant and average kernel weight suggest that a new backcrossing cycle is needed to break eventual undesirable linkages and generate resistant individuals closer to the commercial standards.