Tertiary Gene Pool Research Articles

Natural Diversity in Stomatal Features of Cultivated and Wild Oryza Species

BackgroundStomata in rice control a number of physiological processes by regulating gas and water exchange between the atmosphere and plant tissues. The impact of the structural diversity of these micropores on its conductance level is an important area to explore before introducing stomatal traits into any breeding program in order to increase photosynthesis and crop yield. Therefore, an intensive measurement of structural components of stomatal complex (SC) of twenty three Oryza species spanning the primary, secondary and tertiary gene pools of rice has been conducted.ResultsExtensive diversity was found in stomatal number and size in different Oryza species and Oryza complexes. Interestingly, the dynamics of stomatal traits in Oryza family varies differently within different Oryza genetic complexes. Example, the Sativa complex exhibits the greatest diversity in stomatal number, while the Officinalis complex is more diverse for its stomatal size. Combining the structural information with the Oryza phylogeny revealed that speciation has tended towards increasing stomatal density rather than stomatal size in rice family. Thus, the most recent species (i.e. the domesticated rice) eventually has developed smaller yet numerous stomata. Along with this, speciation has also resulted in a steady increase in stomatal conductance (anatomical, gmax) in different Oryza species. These two results unambiguously prove that increasing stomatal number (which results in stomatal size reduction) has increased the stomatal conductance in rice. Correlations of structural traits with the anatomical conductance, leaf carbon isotope discrimination (∆13C) and major leaf morphological and anatomical traits provide strong supports to untangle the ever mysterious dependencies of these traits in rice. The result displayed an expected negative correlation in the number and size of stomata; and positive correlations among the stomatal length, width and area with guard cell length, width on both abaxial and adaxial leaf surfaces. In addition, gmax is found to be positively correlated with stomatal number and guard cell length. The ∆13C values of rice species showed a positive correlation with stomatal number, which suggest an increased water loss with increased stomatal number. Interestingly, in contrast, the ∆13C consistently shows a negative relationship with stomatal and guard cell size, which suggests that the water loss is less when the stomata are larger. Therefore, we hypothesize that increasing stomatal size, instead of numbers, is a better approach for breeding programs in order to minimize the water loss through stomata in rice.ConclusionCurrent paper generates useful data on stomatal profile of wild rice that is hitherto unknown for the rice science community. It has been proved here that the speciation has resulted in an increased stomatal number accompanied by size reduction during Oryza’s evolutionary course; this has resulted in an increased gmax but reduced water use efficiency. Although may not be the sole driver of water use efficiency in rice, our data suggests that stomata are a potential target for modifying the currently low water use efficiency in domesticated rice. It is proposed that Oryza barthii can be used in traditional breeding programs in enhancing the stomatal size of elite rice cultivars.

Characterization of Cultivated Eggplant and its Wild Relatives Based on Important Fruit Biochemical Traits.

The modern-day eggplant consumers desire varieties with a higher content of chlorogenic acid, but the cultivated varieties of eggplant are with a lower content of chlorogenic acid. Whereas, the wild relatives of eggplant are higher in phenolic acids. Therefore, this study characterized the cultivated eggplant and its wild relatives for the fruit dry matter content, total fruit phenolics and chlorogenic acid content. Fruits of the accessions of cultivated eggplant, one primary genepool species, nine secondary genepool species and three tertiary genepool species were characterized for dry matter content (%), total phenolics and the fruit chlorogenic acid content (mg g-1). The chlorogenic acid content in the fruit flesh was determined by using the High Performance Liquid Chromatography (HPLC). Highest content of dry matter content of around 29% was determined for the species S. tomentosum and S. elaeagnifolium. Whereas, the highest content of total fruit phenolics were determined in the secondary genepool species S. linnaeanum. The most top content of chlorogenic acid around 4.5 mg g-1 of fruit dry weight was present in the species S. linnaeanum and S. torvum. Different clustering approaches were able to cluster the primary genepool species with the cultivated eggplant. Overall, this work provides important information about the wild relatives of eggplant concerning their dry matter content, total phenolics and chlorogenic acid content. This information can be used to engineer eggplant varieties rich in fruit phenolics.

Utilizing Wild Cajanus platycarpus, a Tertiary Genepool Species for Enriching Variability in the Primary Genepool for Pigeonpea Improvement.

The use of crop wild relatives in the breeding program has been well recognized to diversify the genetic base along with introgression of useful traits. Cajanus platycarpus (Benth.) Maesen, an annual wild relative belonging to the tertiary genepool of pigeonpea, possesses many useful traits such as early maturity, high protein content, photoperiod insensitivity, and pod borer tolerance for the genetic improvement of cultivated pigeonpea. Using this cross incompatible wild Cajanus species, an advanced backcross population was developed following the embryo rescue technique. In the present study, a pre-breeding population consisting of 136 introgression lines (ILs) along with five popular varieties (used as checks) was evaluated for important agronomic traits during 2016 and 2017 rainy seasons and for grain nutrient content during 2016, 2017, and 2018 rainy seasons. Large genetic variation was observed for agronomic traits such as days to 50% flowering, number of pods per plant, pod weight per plant, grain yield per plant, and grain nutrients [protein content, grain iron (Fe), zinc (Zn), calcium (Ca), and magnesium (Mg)] in the pre-breeding population. Significant genotype × environment interaction was also observed for agronomic traits as well as grain nutrients indicating the sensitivity of these traits to the environments. No significant correlations were observed between grain yield and grain nutrients except grain Zn content which was negatively correlated with grain yield. Overall, 28 promising high-yielding ILs with high grain nutrient content were identified. These ILs, in particular, ICPP # 171012, 171004, 171102, 171087, 171006, and 171050 flowered significantly earlier than the popular mega variety, ICPL 87119 (Asha) and thus hold potential in developing new short-duration cultivars. The comprehensive multi-site assessment of these high-yielding, nutrient-rich accessions would be useful in identifying region-specific promising lines for direct release as cultivars. Moreover, these ILs are expected to replace the popular existing cultivars or for use as new and diverse sources of variations in hybridization programs for pigeonpea improvement.

Waxy Gene-Orthologs in Wheat × Thinopyrum Amphidiploids

Starch, as the main component of grain in cereals, serves as the major source of calories in staple food and as a raw material for industry. As the technological and digestive properties of starch depend on its content, the management of its components, amylose and amylopectin, is of great importance. The starch properties of wheat grain can be attuned using allelic variations of genes, including granule-bound starch synthase I (GBSS I), or Wx. The tertiary gene pool, including wheatgrass (Thinopyrum) species, provides a wide spectrum of genes-orthologs that can be used to increase the allelic diversity of wheat genes by wide hybridization. Octaploid partial wheat–wheatgrass hybrids (amphidiploids, WWGHs) combine the complete genome of bread wheat (BBAADD), and a mixed genome from the chromosomes of intermediate wheatgrass (Thinopyrum intermedium, genomic composition JrJrJvsJvsStSt) and tall wheatgrass (Th. ponticum, JJJJJJJsJsJsJs). Thus, WWGHs may carry Wx genes not only of wheat (Wx-B1, Wx-A1 and Wx-D1) but also of wheatgrass origin. We aimed to assess the level of amylose in starch and investigate the polymorphism of Wx genes in 12 accessions of WWGHs. Additionally, we characterized orthologous Wx genes in the genomes of wild wheat-related species involved in the development of the studied WWGHs, Th. intermedium and Th. ponticum, as well as in the putative donors of their subgenomes, bessarabian wheatgrass (Th. bessarabicum, JbJb) and bluebunch wheatgrass (Pseudoroegneria stipifolia, St1St1St2St2). Although no significant differences in amylose content were found between different WWGH accessions, SDS-PAGE demonstrated that at least two WWGHs have an additional band. We sequenced the Wx gene-orthologs in Th. bessarabicum, P. stipifolia, Th. intermedium and Th. ponticum, and developed a WXTH marker that can discriminate the Thinopyrum Wx gene in the wheat background, and localized it to the 7E chromosome in Th. elongatum. Using the WXTH marker we revealed the allelic polymorphism of the Thinopyrum Wx gene in the studied WWGHs. The applicability of Thinopyrum Wx genes in wheat breeding and their effect on starch quality are discussed.

Overcoming interspecific barriers in Sorghum and molecular characterization of the derivatives

AbstractCrop improvement in sorghum has progressed extensively availing the variation available in the primary gene pool of S. bicolor. Crossing further, the yield plateau can be accomplished by bringing in the genes from tertiary gene pool into the cultivated gene pool. Earlier efforts for wide hybridization in sorghum with tertiary gene pool were not successful due to inhibition of wild pollen germination on the stigma of cultivated species. In the current study, elite lines of cultivated Sorghum species were pollinated with wild species using different floral treatments. The results showed that spray with the synthetic auxin, 2,4‐dichlorophenoxyacetic acid, has promoted the seed set. Overall, 76% crosses showed seed set, but many seeds were underdeveloped and deformed. F1 progeny showed traits of both cultivated and wild parents. Hybrids were confirmed through PCR analysis. Many (80%) F1 plants were sterile, and 20% of F1s were advanced to F2 generation. Wide variation was observed in F2 progeny, and the phenotype selections were made and are being evaluated in the field.

Evolution and Diversity of the Wild Rice Oryza officinalis Complex, across Continents, Genome Types, and Ploidy Levels.

The Oryza officinalis complex is the largest species group in Oryza, with more than nine species from four continents, and is a tertiary gene pool that can be exploited in breeding programs for the improvement of cultivated rice. Most diploid and tetraploid members of this group have a C genome. Using a new reference C genome for the diploid species O. officinalis, and draft genomes for two other C genome diploid species Oryza eichingeri and Oryza rhizomatis, we examine the influence of transposable elements on genome structure and provide a detailed phylogeny and evolutionary history of the Oryza C genomes. The O. officinalis genome is 1.6 times larger than the A genome of cultivated Oryza sativa, mostly due to proliferation of Gypsy type long-terminal repeat transposable elements, but overall syntenic relationships are maintained with other Oryza genomes (A, B, and F). Draft genome assemblies of the two other C genome diploid species, Oryza eichingeri and Oryza rhizomatis, and short-read resequencing of a series of other C genome species and accessions reveal that after the divergence of the C genome progenitor, there was still a substantial degree of variation within the C genome species through proliferation and loss of both DNA and long-terminal repeat transposable elements. We provide a detailed phylogeny and evolutionary history of the Oryza C genomes and a genomic resource for the exploitation of the Oryza tertiary gene pool.

Characterization of chickpea gene pools for nutrient concentrations under agro-climatic conditions of North-Western Himalayas

AbstractChickpea is one of the most important nutritious grain legume crops in the world. There is limited information available on micro- and macro-nutrients in chickpea. Therefore, an effort was made to evaluate a set of 40 chickpea genotypes belonging to all the gene pools including cultivated (Cicer arietinum) as well as wild, Cicer reticulatum from the primary gene pool, Cicer echinospermum from the secondary gene pool and Cicer microphyllum from the tertiary gene pools. Concentration in the seed of the micro- (Zn, Fe, Cu and Mn) and macro-nutrients (Ca, Mg and K) was studied. Substantial variation was observed among different gene pools for the concentration of all the nutrients. The cultivated chickpea exhibited higher seed Cu, Mn, Mg and Ca than wilds indicating positive domestication effect, whereas wild crop relatives were found to have higher levels than cultivated chickpeas for seed Zn, Fe and K concentrations. While comparing desi-type chickpeas with Kabulis, it was revealed that desi types possessed more Zn, Cu, Ca and Mg than Kabulis but reverse was true for seed Fe, Mn and K. Among different desi types (desi brown, desi green and desi black), desi brown types were generally associated with higher mineral nutrient levels. The present study led to the identification of most promising genotypes for different seed micro- and macro-nutrients. These promising lines may serve as genetic resources useful in gene discovery programmes and for alleviating malnutrition or hidden hunger in the developing world.

Genome mapping of quantitative trait loci (QTL) controlling domestication traits of intermediate wheatgrass (Thinopyrum intermedium).

Allohexaploid (2n = 6x = 42) intermediate wheatgrass (Thinopyrum intermedium), abbreviated IWG, is an outcrossing perennial grass belonging to the tertiary gene pool of wheat. Perenniality would be valuable option for grain production, but attempts to introgress this complex trait from wheat-Thinopyrum hybrids have not been commercially successful. Efforts to breed IWG itself as a dual-purpose forage and grain crop have demonstrated useful progress and applications, but grain yields are significantly less than wheat. Therefore, genetic and physical maps have been developed to accelerate domestication of IWG. Herein, these maps were used to identify quantitative trait loci (QTLs) and candidate genes associated with IWG grain production traits in a family of 266 full-sib progenies derived from two heterozygous parents, M26 and M35. Transgressive segregation was observed for 17 traits related to seed size, shattering, threshing, inflorescence capacity, fertility, stem size, and flowering time. A total of 111 QTLs were detected in 36 different regions using 3826 genotype-by-sequence markers in 21 linkage groups. The most prominent QTL had a LOD score of 15 with synergistic effects of 29% and 22% over the family means for seed retention and percentage of naked seeds, respectively. Many QTLs aligned with one or more IWG gene models corresponding to 42 possible domestication orthogenes including the wheat Q and RHT genes. A cluster of seed-size and fertility QTLs showed possible alignment to a putative Z self-incompatibility gene, which could have detrimental grain-yield effects when genetic variability is low. These findings elucidate pathways and possible hurdles in the domestication of IWG.

Characterization of host response to bruchids (Callosobruchus chinensis and C. maculatus) in 39 genotypes belongs to 12 Cajanus spp. And assessment of molecular diversity inter se

Thirty nine genotypes belong to 12 Cajanus species were characterized for host response to bruchids (C. chinensis and C. maculatus) using no-choice based in vivo and in vitro antibiosis assay, and these assays revealed seven genotypes of primary gene pool (C. cajan) were more susceptible to bruchid infestation as compared to the genotypes of secondary and tertiary gene pools. Among all the genotypes, C. cajanifolius acc. ICPW-31 showed higher degree of resistance to both the bruchids. The molecular diversity using CDDP and SCoT markers showed wide range of genetic variations among the 12 species of Cajanus, and was supported by estimates of Nei's genetic distance and fixation index (FST) based haplotype matrix. The genetic structure showed clustering of 39 genotypes into eight distinct groups (K = 8) on the basis of their allelic composition, and among them C. cajanifolius acc. ICPW 31 and ICPW 30 showed close affinity with the cultivars of C. cajan by sharing several alleles. AMOVA analyses showed the existence of higher extent of genetic variation both at the genotype (37%) and species (63%) level in the genus Cajanus. The dendrogram and the principal coordinate analysis (PCoA) placed the 39 genotypes into six major clusters at par with their sectional classification. MCheza and ARLEQUIN based outlier analysis revealed 13 common loci under balancing selection, which are supposed to be involved in maintenance of genetic polymorphism either at species or genotype level, and are assigned to putative gene families such as KNOX, WRKY, ERF, MYB, ABP1 and MADS. The in vivo and in vitro bruchid assay vis-à-vis the DNA marker based molecular diversity analysis affirmed the possible use of C. cajanifolius acc. ICPW-31 as donor genotype for the introgression of bruchid resistance allele(s) into cultivated genetic background.

The oat gene pools – review about the use of wild species in improving cultivated oat

The search for agronomic traits and the use of new sources of variability in oat farming is very important in terms of breeding. Wild species of Avena are grouped into three gene pools depending on their interfertility with cultivated hexaploid oat. The primary and tertiary gene pools are extensive and diverse, the secondary gene pool is relatively small and poorly represented in ex situ gene banks. Appropriate wild species are a valuable source of many appropriate traits such as: high protein, oil, ß-glucan and balanced amino acids composition contents in grain; short culm; cold and drought tolerance. Moreover they are a source of resistance genes for oat diseases, such as powdery mildew, crown and stem rust, smuts or BYDV. Note, however transfer of genes from wild to cultivated species is a long and laborious process. These types of methods are used in the species Avena with various end effects. The purpose of this article was to collect information on attempts of transferring different genes from wild oat species to cultivated oats.

Identification of New Sources of Resistance to Wheat Stem Rust in Aegilops spp. in the Tertiary Genepool of Wheat.

Recent stem rust epidemics in eastern Africa and elsewhere demonstrated that wheat stem rust is a re-emerging disease posing a threat to wheat production worldwide. The cultivated wheat gene pool has a narrow genetic base for resistance to virulent races, such as races in the Ug99 race group. Wild relatives of wheat are a tractable source of stem rust resistance genes. Aegilops species in the tertiary genepool have not been exploited to any great extent as a source of stem rust resistance. We evaluated 1,422 accessions of Aegilops spp. for resistance to three highly virulent races (TTKSK, TRTTF, and TTTTF) of Puccinia graminis f. sp. tritici. Species studied include Ae. biuncialis, Ae. caudata, Ae. comosa, Ae. cylindrica, Ae. geniculata, Ae. neglecta, Ae. peregrina, Ae. triuncialis, and Ae. umbellulata that do not share common genomes with cultivated wheat. High frequencies of resistance were observed as 977 (68.8%), 927 (65.2%), and 850 (59.8%) accessions exhibited low infection types to races TTKSK, TTTTF, and TRTTF, respectively. Contingency table analyses showed strong association for resistance to different races in several Aegilops spp., indicating that for a given species, the resistance genes effective against multiple races. Inheritance studies in selected accessions showed that resistance to race TTKSK is simply inherited.

Genetic relationships and structured diversity of Lactuca georgica germplasm from Armenia and the Russian Federation among other members of Lactuca L., subsection Lactuca L., assessed by TRAP markers

We studied the genetic relationships of Lactuca georgica samples originating in Armenia and the Russian Federation with samples representing four other predominantly self-pollinating wild Lactuca species (L. serriola, L. aculeata, L. saligna, and L. virosa) originating in various countries, as well as with samples representing cultivated lettuce, L. sativa by using 48 TRAP markers. We also visualized their genetic diversity and structure. The present study is likely the first molecular phylogenetic evaluation of a detailed screening of L. georgica germplasm. Data analysis of the three major wild species in this study, L. georgica (134 samples), L. virosa (57 samples), and L. serriola (40 samples) showed that allele frequencies of all 47 polymorphic loci varied significantly among the species. A total of 11, 9, and 10 alleles were unique to L. georgica, L. serriola, and L. virosa, respectively; 71% of TRAP marker diversity was between species. The Neighbor-Joining tree clearly clustered the whole set of 238 samples according to their taxonomic determination. It also reflects the gene diversity as well as the genetic distance values among samples representing the between and within variance of the various species. The L. georgica samples clustered most distantly from the L. sativa samples. The interspecies comparisons between samples belonging to L. georgica with those belonging to L. sativa displayed a high distance, lower only from the interspecies comparisons between samples belonging to L. virosa (in the tertiary gene pool of cultivated lettuce) with those belonging to L. sativa. Thus, additional molecular data with more hybridization experiments are necessary to reconsider if L. georgica is indeed a constituent of the primary gene pool of cultivated lettuce. The L. georgica samples were divided into two sub-clusters, with samples collected in southeast and central Armenia grouping together while all those collected in the north and Dagestan grouped together.

Genetic Introgression from Glycine tomentella to Soybean to Increase Seed Yield

Soybean [Glycine max (L.) Merr.] breeding in the United States currently relies on a narrow genetic base. Glycine tomentella Hayata (2n = 78), native to Australia, is a perennial relative in the tertiary gene pool of soybean. No effort has been devoted to using this species to increase seed yield. The objectives of this research were (i) to identify high‐yielding lines derived from backcrosses between the soybean cultivar Dwight (2n = 40) and G. tomentella PI 441001 (2n = 78) and to compare their agronomic performance with that of the recurrent parent, and (ii) to determine associations between G. tomentella introgressions and agronomic traits. PI 441001 was crossed to Dwight and immature seed rescue was used to produce a sterile F1 plant. Amphidiploid plants (2n = 118) were produced by treating the F1 hybrid with colchicine. Amphidiploid plants were backcrossed to Dwight and a series of backcrosses were made to obtain lines with 2n = 40 chromosomes, which were self‐fertile and genetically stable. Preliminary yield testing was used to select 180 lines in maturity groups II, III, and IV that had yields greater than or less than that of the recurrent parent. Yield data collected in two replication tests at six to eight locations in 2013 and 2014 identified experimental lines in all three maturity groups that were higher yielding than the recurrent parent. All experimental and parental lines were genotyped, revealing an average of 1% G. tomentella introgression. Significant associations were detected between G. tomentella introgressions and time of flowering, height, lodging, and yield.

Seed protein fraction electrophoresis in peanut (Arachis hypogaea L.) accessions and wild species

Total seed storage proteins were studied in 50 accessions of A. hypogaea (11 A. hypogaea ssp. hypogaea var hypogaea, 13 A. hypogaea ssp. hypogaea var hirsuta, 11 A. hypogaea ssp. fastigiata var fastigiata and 15 A. hypogaea ssp. fastigiata var. vulgaris accessions) in SDS PAGE. These accessions were also analysed for albumin and globulin seed protein fractions. Among the six seed protein markers presently used, it was found that globulin fraction showed maximum diversity (77.2%) in A. hypogaea accessions followed by albumin (52.3%), denatured total soluble protein fraction in embryo (33.3%) and cotyledon (28.5%). The cluster analysis based on combined data of cotyledons, embryos, albumins and globulins seed protein fractions demarcated the accessions of two subspecies hypogaea and fastigiata into two separate clusters supported by 51% bootstrap value, with few exceptions, suggesting the genotypes to be moderately diverse. Native and denatured total soluble seed storage proteins were also electrophoretically analysed in 27 wild Arachis species belonging to six sections of the genus. Cluster analysis using different methods were performed for different seed proteins data alone and also in combination. Section Caulorrhizae (C genome) and Triseminatae (T genome) formed one, distantly related group to A. hypogaea and other section Arachis species in the dendrogram based on denatured seed storage proteins data. The present analysis has maintained that the section Arachis species belong to primary and secondary genepools and, sections Procumbenetes and Erectoides belong to tertiary gene pools.

Efficient anchoring of alien chromosome segments introgressed into bread wheat by new Leymus racemosus genome-based markers

BackgroundThe tertiary gene pool of bread wheat, to which Leymus racemosus belongs, has remained underutilized due to the current limited genomic resources of the species that constitute it. Continuous enrichment of public databases with useful information regarding these species is, therefore, needed to provide insights on their genome structures and aid successful utilization of their genes to develop improved wheat cultivars for effective management of environmental stresses.ResultsWe generated de novo DNA and mRNA sequence information of L. racemosus and developed 110 polymorphic PCR-based markers from the data, and to complement the PCR markers, DArT-seq genotyping was applied to develop additional 9990 SNP markers. Approximately 52% of all the markers enabled us to clearly genotype 22 wheat-L. racemosus chromosome introgression lines, and L. racemosus chromosome-specific markers were highly efficient in detailed characterization of the translocation and recombination lines analyzed. A further analysis revealed remarkable transferability of the PCR markers to three other important Triticeae perennial species: L. mollis, Psathyrostachys huashanica and Elymus ciliaris, indicating their suitability for characterizing wheat-alien chromosome introgressions carrying chromosomes of these genomes.ConclusionThe efficiency of the markers in characterizing wheat-L. racemosus chromosome introgression lines proves their reliability, and their high transferability further broadens their scope of application. This is the first report on sequencing and development of markers from L. racemosus genome and the application of DArT-seq to develop markers from a perennial wild relative of wheat, marking a paradigm shift from the seeming concentration of the technology on cultivated species. Integration of these markers with appropriate cytogenetic methods would accelerate development and characterization of wheat-alien chromosome introgression lines.

Chromosome Rearrangements Caused by Double Monosomy in Wheat-Barley Group-7 Substitution Lines

Interspecific or introgressive hybridization is one of the driving forces in plant speciation, producing allopolyploids or diploids with rearranged genomes. The process of karyotype reshaping following homoploid interspecific hybridization has not been studied experimentally. Interspecific hybridization is widely used in plant breeding to increase genetic diversity and introgress new traits. Numerous introgression stocks were developed for hexaploid wheat Triticum aestivum L. (2n = 6x = 42, genome AABBDD). Double monosomic lines, containing one alien chromosome from the tertiary gene pool of wheat and one homoeologous wheat chromosome, represent a simplified model for studying chromosome rearrangements caused by interspecific hybridization. The pairing of a chromosome from the tertiary gene pool with a wheat homoeologue is restricted by the activity of the wheat Ph1 gene, thus, rearrangements caused by chromosome breakage followed by the fusion of the broken arms can be expected. We analyzed chromosome aberrations in 4 sets of lines that originated from double monosomics of barley (Hordeum vulgare L.) chromosome 7H and wheat group-7 chromosomes with dicentric or ring chromosomes. The dynamics of wheat-barley dicentric chromosomes during plant development was followed and an increased diversity of rearrangements was observed. Besides the targeted group-7 chromosomes, other wheat chromosomes were involved in rearrangements, as chromosomes broken in the centromeric region fused with other broken chromosomes. In some cells, multi-centric chromosomes were observed. The structure and dosage of the introgressed barley chromatin was changed. The transmission of the rearrangements to the progenies was analyzed. The observed aberrations emphasize the importance of cytogenetic screening in gene introgression projects.

Inducement and identification of chromosome introgression and translocation of Gossypium australe on Gossypium hirsutum

BackgroundWe previously reported the development of a set of Gossypium hirsutum-G. australe alien chromosome addition lines. Naturally, however, G. hirsutum-G. australe chromosome exchanges were very limited, impeding the stable transference of useful genes from G. australe (G2G2 genome) into the most cultivated cotton, G. hirsutum (AADD).ResultsIn the present report, the pollen from a pentaploid (2n = AADDG2) of G. hirsutum-G. australe was irradiated with seven different doses ranging from 10 to 40 Grays and used to pollinate emasculated flowers of G. hirsutum over three consecutive years. Irradiation greatly increased the genetic recombination rates of the G. hirsutum and G. australe chromosomes and a total of 107 chromosome introgression individuals in 192 GISH-negative (with no GISH signal on chromosome) survived individuals, 11 chromosome translocation individuals (containing 12 chromosome translocation events) and 67 chromosome addition individuals were obtained in 70 GISH-positive (with GISH signal(s) on chromosome(s)) survived individuals, which are invaluable for mining desirable genes from G. australe. Multicolor genomic in situ hybridization results showed that there were three types of translocation, whole arm translocation, large alien segment translocation and small alien segment translocation, and that all translocations occurred between the G2-genome and the A-subgenome chromosomes in G. hirsutum. We also found that higher doses induced much higher rates of chromosome variation but also greatly lowered the seed viability and seedling survivability.ConclusionsIrradiation has been successfully employed to induce chromosome introgressions and chromosome translocations and promote chromosome exchanges between cultivated and wild species. In addition, by balancing the rates of chromosome introgression and translocation to those of seed set, seed germination, and seedling rates in the M1 generation, we conclude that the dosage of 20 Grays is the most suitable. The established methodology may guide the utilization of the tertiary gene pool of Gossypium species such as G. australe in cotton breeding in the future.

Development of pollen germination medium to test pollen viability of eggplant and its wild species

In this study, we report in vitro pollen germination medium (PGM) for the egg plant and four of its wild species belonging to tertiary gene pool viz., Solanum torvum, S. khasianum, S. trilobatum and S. surrettence. The medium developed by Brewbaker and Kwack has been modified and improved with addition of poly ethylene glycol, ɛ-amino caproic acid etc which supported over 78% pollen germination and pollen tube growth of these species. The complete pollen germination medium for eggplant consists of 20% maltose + 250 mg l−1 Boric acid + 300 mg l−1 Calcium nitrate + 15% PEG 4000 + 750 mg l−1 EACA + 0.5–1% agar at 25 oC temperature for 3h of incubation. During the study, pollen sterility of 8–12% was observed at flowering period invariably in all Solanum species.

Development and Molecular Characterization of Wheat-Aegilops peregrina Introgression Lines with Resistance to Leaf Rust and Stripe Rust

ABSTRACTA wild non-progenitor species from wheat (Triticum aestivum L.) tertiary gene pool, Aegilops peregrina (Hack.) Maire & Weiller accession pau3519 (UUSS), was used for introgression of leaf rust (caused by Puccinia triticina) and stripe rust (caused by Puccinia striiformis f. sp. tritici) resistance in bread wheat. The accession was crossed and backcrossed with hexaploid wheat line Chinese Spring PhI to develop two homozygous BC2F6 wheat-Ae. peregrina introgression lines (ILs), viz., IL pau16058 and IL pau16061, through induced homoeologous recombination. Homozygous lines were screened against six Puccinia triticina and two Puccinia striiformis f. sp. tritici pathotypes at the seedling stage and a mixture of prevalent pathotypes of both species at the adult plant stage. IL pau16061 showed resistance to leaf rust only, whereas IL pau16058 was resistant to both leaf and stripe rust pathotypes throughout plant life. Molecular profiling of these ILs with simple sequence repeat (SSR) markers indicated that alien introgressions were mainly terminal and very few were interstitial. Identification of linked markers with advanced genomic technologies will aid in marker-assisted pyramiding of alien genes in cultivated wheat background.

Prolific triploid production in intersectional crosses of 4x Vaccinium corymbodendron Dunal (section Pyxothamnus) by 2x section Cyanococcus species

Vaccinium corymbodendron (section Pyxothamnus) is a tetraploid species native to high-altitude locations in Peru, Colombia, and Venezuela, and is of considerable interest because it flowers at times when nighttime temperatures drop below freezing. It also is notable for its profuse, concentrated flowering, and monopodial plant structure both of which may be useful in breeding for mechanical harvest. Initial hybridization experiments of 4x V. corymbodendron with a range of 2x species from section Cyanococcus found all hybrids were triploids. 4x × 2x crosses in conventional Vaccinium germplasm typically result in an almost absolute block of triploid offspring. The triploids expressed very low fertility as both males and females, nonetheless, several were used successfully in crosses. A single hybrid from 4x V. corymbodendron × 2x V. vitis-idaea L. (lingonberry, section Vitis-idaea) was unexpectedly found to be tetraploid and was highly fertile. These crossing results suggest V. corymbodendron either possesses no ploidy barriers to hybridization, or possesses genomic dosage factors that differ from section Cyanococcus species. Understanding these crossing relationships may open new avenues for exploitation of this germplasm and other tertiary gene pool germplasm.