Origin Of Germplasm Research Articles

Origin of current intermediate wheatgrass germplasm being developed for Kernza grain production

AbstractIntermediate wheatgrass (IWG, Thinopyrum intermedium [Host] Barkworth & D. R. Dewey) has been developed as a perennial grain crop for human consumption along with providing environmental benefits and ecosystem services. Grain and products derived from IWG cultivars improved for food production have been marketed under the registered trademark, Kernza. Development of IWG as a perennial grain crop began in 1980s with a phenotypic recurrent selection program as the Rodale Institute (RI) and the Big Flats Plant Material Center (BFPMC) used IWG plant introductions (PI) from the National Plant Germplasm System (NPGS) to improve populations of IWG. Initial selections were provided to The Land Institute (TLI) where they were subsequently improved for grain production, yet the identity of the founder material of improved, food-grade IWG has not been publicly documented. Recently recovered original documents have been used to reconstruct the early breeding program to identify the most likely 20 PIs that form the founders of modern food-grade IWG. Molecular data using genotyping-by-sequencing in current elite breeding material, and remnant seed and plant material from the initial RI selections have provided supporting evidence for the historical records. The genetic origin for food-grade IWG is focused between the Black Sea and Caspian Sea in the Stavropol region of Russia, with smaller contributions likely from collections as distant as Kazakhstan in the east to Turkey in the west. This work connects the flow of germplasm and utility of NPGS PIs to present day IWG grain cultivars being developed in multiple breeding programs around the world.

Developing EST-SSR Markers for Identifying and Evaluating Asparagus Germplasm Resources Based on Transcriptome Sequences

Radix asparagi is the dried root tuber of the Liliaceae plant Asparagus cochinchinensis (Lour.) Merr., which is a major Chinese medicinal herb with high medicinal and edible value in China. The planting area of A. cochinchinensis is extensive, and there is blind introduction in various regions, leading to confusion the origin of Radix asparagi and impure germplasm. This study conducted morphological and karyotype analyses on cultivated Asparagus resources from seven main production areas in China and developed SSR molecular markers suitable for the identification of Asparagus germplasm resources based on the transcriptome sequencing results. The morphological results indicate that in addition to A. cochinchinensis (Lour.) Merr., recorded in the Pharmacopoeia of the People’s Republic of China, there are also A. taliensis Wang et Tang and A. lycopodineus (Baker) Wang et Tang cultivated in China. All the tested Asparagus resources were diploid and had 20 chromosomes. A total of 8841 single genes containing SSR loci were identified using transcriptome sequencing of Neijiang Asparagus, including 761 SSR loci with trinucleotide repeat units. One hundred pairs of SSR primers were randomly designed from the trinucleotide repeat loci for PCR and polymorphism verification, and ten pairs were selected for identification of Asparagus germplasm resources. The genetic diversity results of ten pairs of primers in seven Asparagus-producing regions were consistent with the morphological identification. This study provides technical support for the comprehensive evaluation and utilization of Asparagus germplasm resources.

Diversity, classification, and EST‐SSR‐based association analysis of caladium ornamental traits

Caladium (Caladium × Hortulanum Birdsey) is a popular ornamental plant with a wide range of vibrant leaf color among Araceae. Even after years of breeding, creating new caladium leaf color variations is extremely difficult. Molecular marker-assisted selection is an effective approach for accelerating breeding, but few studies on the molecular markers associated with caladium traits have been performed. In the current study, 144 caladium accessions were used to examine 12 phenotypic characteristics. The coefficient of variation for four numerical characters ranged from 23.94% to 43.22%, and the Shannon-Wiener indexes for eight descriptive characters ranged from 0.13 to 1.52. Based on L*, a*, b*, C, h° values determined by a colorimeter and hierarchical cluster analysis, the leaf color can be divided into four groups: pale green, green, light pink, and red. Furthermore, 7708 new SSR loci were identified by transcriptome sequencing, and 26 SSR markers with high polymorphism and reproducibility were screened. Genetic structure, NJ clustering, and PCoA analysis revealed that 144 accessions could be divided into three clusters, with genetic structure being closely related to germplasm origin. An association analysis revealed that the SSR markers 2, 1, 1, 1, 1, and 1 were mainly associated with petiole color, main vein color, blade upperside glossiness, and C, b*, and L* of leaf color (p < 0.01). These findings will serve as a valuable reference for evaluating germplasm resources and caladium molecular marker-assisted breeding.

Natural variations and geographical distributions of seed carotenoids and chlorophylls in 1 167 Chinese soybean accessions

Understanding the composition and contents of carotenoids in various soybean seed accessions is important for their nutritional assessment. This study investigated the variability in the concentrations of carotenoids and chlorophylls and revealed their associations with other nutritional quality traits in a genetically diverse set of Chinese soybean accessions comprised of cultivars and landraces. Genotype, planting year, accession type, seed cotyledon color, and ecoregion of origin significantly influenced the accumulation of carotenoids and chlorophylls. The mean total carotenoid content was in the range of 8.15–14.72 µg g–1 across the ecoregions. The total carotenoid content was 1.2-fold higher in the landraces than in the cultivars. Soybeans with green cotyledons had higher contents of carotenoids and chlorophylls than those with yellow cotyledons. Remarkably, lutein was the most abundant carotenoid in all the germplasms, ranging from 1.35–37.44 µg g–1. Carotenoids and chlorophylls showed significant correlations with other quality traits, which will help to set breeding strategies for enhancing soybean carotenoids without affecting the other components. Collectively, our results demonstrate that carotenoids are adequately accumulated in soybean seeds, however, they are strongly influenced by genetic factors, accession type, and germplasm origin. We identified novel germplasms with the highest total carotenoid contents across the various ecoregions of China that could serve as the genetic materials for soybean carotenoid breeding programs, and thereby as the raw materials for food sectors, pharmaceuticals, and the cosmetic industry.

Development of single nucleotide polymorphism markers and genetic diversity in guava (Psidium guajava L.)

Societal impact statementThere are many understudied fruits of great economic and productive importance for regional agricultural hubs. Guava is an American fruit species with attractive nutritional and adaptability characteristics. However, in many countries like Mexico, its cultivation is still limited to small‐to‐medium size plantings and home gardens and depends on poorly controlled germplasm that diminishes productivity and expansion. Our study provides valuable insight to better understand guava diversity and generate high‐yielding, high‐quality, and better adapted materials. Supporting the study and development of understudied crops will provide us with more resources for facing climate change adversities and for diversifying cropping systems.Summary Guava (Psidium guajava L.) is a fruit crop species native to tropical and subtropical regions of America with great productive and economic potential due to its extensive environmental adaptability, nutritional value, and medicinal properties. However, the lack of molecular resources for accelerated breeding, limited knowledge about its evolutionary and domestication history, and unfavorable policies have limited its genetic improvement and broader adoption as a commercial fruit crop. Here, we present the first diversity study in guava employing genome‐wide single nucleotide polymorphisms (SNPs). Forty‐eight accessions collected from Mexico and other continents were examined with more than 6000 high‐quality SNP markers, which represents a marker density increase of ~30x compared with previous studies. Relationships between genetic groupings and geographic origins were not apparent in this diverse guava collection using principal component and structure analyses. Extensive germplasm exchange among guava‐producing regions and limited varietal control at commercial plantations might have contributed to ambiguities when defining the true origin and identities of existing germplasm materials. However, an analysis of domestication syndrome traits (fruit size and sugar) by wild or improved germplasm revealed several putative genomic regions under selection. Knowledge about germplasm origins and genetic relatedness, in conjunction with reliable molecular resources and better agronomic practices, is necessary to support breeding efforts and facilitate broader adoption of orphan crops such as guava, which have increasingly important roles in light of climate change adversities and in diversifying diets and food systems.

Origin, Maturity Group and Seed Coat Color Influence Carotenoid and Chlorophyll Concentrations in Soybean Seeds.

Soybean (Glycine max (L.) Merrill) seeds are abundant in physiologically active metabolites, including carotenoids and chlorophylls, and are used as an affordable source of functional foods that promote and maintain human health. The distribution and variation of soybean seed metabolites are influenced by plant genetic characteristics and environmental factors. Here, we investigated the effects of germplasm origin, genotype, seed coat color and maturity group (MG) on the concentration variation of carotenoid and chlorophyll components in 408 soybean germplasm accessions collected from China, Japan, the USA and Russia. The results showed that genotype, germplasm origin, seed color, and MG were significant variation sources of carotenoid and chlorophyll contents in soybean seeds. The total carotenoids showed about a 25-fold variation among the soybean germplasms, with an overall mean of 12.04 µg g−1. Russian soybeans yielded 1.3-fold higher total carotenoids compared with Chinese and Japanese soybeans. Similarly, the total chlorophylls were substantially increased in Russian soybeans compared to the others. Soybeans with black seed coat color contained abundant concentrations of carotenoids, with mainly lutein (19.98 µg g−1), β-carotene (0.64 µg g−1) and total carotenoids (21.04 µg g−1). Concentrations of lutein, total carotenoids and chlorophylls generally decreased in late MG soybeans. Overall, our results demonstrate that soybean is an excellent dietary source of carotenoids, which strongly depend on genetic factors, germplasm origin, MG and seed coat color. Thus, this study suggests that soybean breeders should consider these factors along with environmental factors in developing carotenoid-rich cultivars and related functional food resources.

Assessment of Phenotypic Diversity in the USDA Collection of Quinoa Links Genotypic Adaptation to Germplasm Origin.

Quinoa’s germplasm evaluation is the first step towards determining its suitability under new environmental conditions. The aim of this study was to introduce suitable germplasm to the lowland areas of the Faisalabad Plain that could then be used to introduce quinoa more effectively to that region. A set of 117 quinoa genotypes belonging to the USDA quinoa collection was evaluated for 11 phenotypic quantitative traits (grain yield (Y), its biological and numerical components plus phenological variables) in a RCBD during two consecutive growing seasons at the University of Agriculture, Faisalabad, Pakistan under mid-autumn sowings. Genotypic performance changed across the years, however most phenotypic traits showed high heritability, from 0.75 for Harvest Index (HI) to 0.97 for aerial biomass (B) and Y. Ordination and cluster analyses differentiated four groups dominated by genotypes from: Peru and the Bolivian Highlands (G1); the Bolivian Highlands (G2); the Ballón collection (regarded as a cross between Bolivian and Sea Level (Chilean) genotypes) plus Bolivian Highlands (G3); and Ballón plus Sea Level (G4), this latter group being the most differentiated one. This genetic structure shared similarities with previous groups identified using SSR markers and G×E data from an international quinoa test. G4 genotypes showed the highest Y associated with higher B and seed numbers (SN), while HI made a significant contribution to yield determination in G2 and seed weight (SW) in G3. G1 and G2 showed the lowest Y associated with a lower B and SN. Moreover, SW showed a strongly negative association with SN in G2. Accordingly, G4 followed by G3 are better suited to the lowland areas of Faisalabad plain and the physiological traits underlying yield determination among genotypic groups should be considered in future breeding programs.

Genetic Diversity Analysis of Indonesian Pigmented Rice Accessions Based on Functional Molecular Markers

Pigmented rice is functional staple food that becomes popular because of its anthocyanin content which is beneficial for health. Studies on the diversity of the local variety of Indonesian pigmented rice accessions have been carried out, but are still limited to one region of germplasm origin. This study aimed to analyze the genetic diversity of local varieties of pigmented rice collections of the IAARD-ICABIOGRAD Gene Bank. A total of 93 pigmented rice accessions from 16 provinces in Indonesia were analyzed using 15 functional molecular markers of SSR, STS, and indel. The total alleles detected were 115 with an average per locus of genetic diversity value of 0.71. There were five markers with PIC values &amp;gt;0.75, i.e. RM167, RM223, R8M33, R10M10, and GBSS1. The accessions were divided into two main groups based on their pericarp color. It is necessary to analyze the physicochemical content of the local rice accessions to complement the existing diversity information and identify potential pigmented rice accessions with high palatability.

Genome-wide association studies of leaf angle in maize

Compact plant-type with small leaf angle has increased canopy light interception, which is conducive to the photosynthesis of the population and higher population yield at high density planting in maize. In this study, a panel of 285 diverse maize inbred lines genotyped with 56,000 SNPs was used to investigate the genetic basis of leaf angle across 3 consecutive years using a genome-wide association study (GWAS). The leaf angle showed broad phenotypic variation and high heritability across different years. Population structure analysis subdivided the panel into four subgroups that correspond to the four major empirical germplasm origins in China, i.e., Tangsipingtou, Reid, Lancaster and P. When tested with the optimal GWAS model, we found that the Q + K model was the best in reducing false positive. In total, 96 SNPs accounting for 5.54–10.44% of phenotypic variation were significantly (P < 0.0001) associated with leaf angle across three years. According to the linkage disequilibrium decay distance, 96 SNPs were binned into 43 QTLs for leaf angle. Seven major QTLs with R2 > 8% stably detected in at least 2 years, and BLUP values were clustered in four genomic regions (bins 2.01, 2.07, 5.06, and 10.04). Seven important candidate genes, Zm00001d001961, Zm00001d006348, Zm00001d006463, Zm00001d017618, Zm00001d024919, Zm00001d025018, and Zm00001d025033 were predicted for the seven stable major QTLs, respectively. The markers identified in this study can be used for molecular breeding for leaf angle, and the candidate genes would contribute to further understanding of the genetic basis of leaf angle.

Resistance to Meloidogyne enterolobii and Meloidogyne incognita in Cultivated and Wild Cowpea

Cowpea [Vigna unguiculata (L.) Walp] is a multipurpose crop that provides nutrients for human and livestock diets, as well as regulates and supports ecosystem services. In developing countries, cowpea is exploited as a dual-purpose crop for its grain and fodder; it is cultivated primarily for grain and as a cover crop in industrialized countries. However, root-knot nematodes (RKNs) (Meloidogyne spp.) represent a threat to cowpea production worldwide. Thus, we screened the University of California, Riverside (UC-Riverside), cowpea mini-core collection for resistance to Meloidogyne incognita Kofoid and White (Chitwood) and M. enterolobii Yang and Eisenback to verify the potential of this collection to be used for improving RKN resistance in cowpeas. Both screenings showed significant genotypic variation and medium/high broad-sense heritability (H2) estimates for most traits, and several traits were also strongly correlated. For the M. incognita screening, 86.1% of accessions showed some level of resistance based on gall score (≤3), and 77.7% based on reproduction index (RI) (25 ≤ RI ≤ 50), whereas only 10.4% and 29.8% of accessions were resistant to M. enterolobii based on gall score (≤3) and RI (25 ≤ RI ≤ 50), respectively. These results demonstrate the greater virulence of M. enterolobii than M. incognita in cowpea, and that geographic origin of germplasm was not linked to sources of resistance. Among cultivars, only US-1136 showed resistance against both nematode species, whereas 12 wild/landrace germplasms exhibited resistance to M. incognita and M. enterolobii, and can be exploited for breeding resistant cowpeas.

Parentage Atlas of Italian Grapevine Varieties as Inferred From SNP Genotyping.

The Italian grape germplasm is characterized by a high level of richness in terms of varieties number, with nearly 600 wine grape varieties listed in the Italian National Register of Grapevine Varieties and with a plethora of autochthonous grapes. In the present study an extended SNP genotyping has been carried out on Italian germplasm of cultivated Vitis vinifera subsp. sativa and Vitis hybrids. Several hundred Italian varieties maintained in the repositories of scientific Institutions and about one thousand additional varieties derived from previous studies on European, Southern Italy, Magna Graecia and Georgian germplasm were considered. The large genotyping data obtained were used to check the presence of homonyms and synonyms, determine parental relationships, and identify the main ancestors of traditional Italian cultivars and closely-related accessions. The parentage among a set of 1,232 unique varieties has been assessed. A total of 92 new parent-offspring (PO) pairs and 14 new PO trios were identified. The resulted parentage network suggested that the traditional Italian grapevine germplasm originates largely from a few central varieties geographically distributed into several areas of genetic influence: “Strinto porcino” and its offspring “Sangiovese”, “Mantonico bianco” and “Aglianico” mainly as founder varieties of South-Western Italy (IT-SW); Italian Adriatic Coast (IT-AC); and Central Italy with most varieties being offsprings of “Visparola”, “Garganega” and “Bombino bianco”; “Termarina (Sciaccarello)” “Orsolina” and “Uva Tosca” as the main varieties of North-Western Italy (IT-NW) and Central Italy. The pedigree reconstruction by full-sib and second-degree relationships highlighted the key role of some cultivars, and, in particular, the centrality of “Visparola” in the origin of Italian germplasm appeared clear. An hypothetical migration of this variety within the Italian Peninsula from South to North along the eastern side, as well as of “Sangiovese” from South to Central Italy along the Western side might be supposed. Moreover, it was also highlighted that, among the main founders of muscat varieties, “Moscato bianco” and “Zibibbo (Muscat of Alexandria)” have spread over the whole Italy, with a high contribution by the former to germplasm of the North-Western of the peninsula.

Characterization of phytochemicals in the roots of wild herbaceous peonies from China and screening for medicinal resources

Paeonia Radix Rubra (PRR) is a very common traditional Chinese medicine (TCM). The roots of Paeonia lactiflora and Paeonia anomala subsp. veitchii are used for the production of PRR. However, other species of section Paeonia in China are also used to produce PRR. The roots of section Paeonia from 20 populations of seven species and two subspecies in China were analyzed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF- MS). A total of 21 metabolites were identified, including nine monoterpene glycosides, seven tannins, three phenols, one paeonol and one flavonoid. There were significant differences in the composition and content of metabolites among different populations. The relative contents of monoterpene glycosides and tannins were generally higher in most samples. Cluster analysis showed that 20 populations could be divided into four groups. Among them, the populations of P. lactiflora and Paeonia mairei were clustered together in one group. The content of paeoniflorin in P. lactiflora was high (>22.20 mg g−1, dry weight, the same below), and the content of lactiflorin in P. mairei (>17.50 mg g−1) was significantly higher than in other species (<2.30 mg g−1). The monoterpene glycoside content in P. mairei (>51.60 mg g−1) was significantly higher than in other species (<43.40 mg g−1), suggesting that it could be useful medicinal germplasm for the development of monoterpene glycosides. In addition, some populations of Paeonia emodi, Paeonia sterniana and P. mairei may also be used as potential germplasm resources for use in PRR. Genetic and environmental factors resulted in differences in the composition and content of metabolites in different populations of the same species. Therefore, it is necessary to carefully consider the origin of Paeonia germplasm when selecting medicinal materials to be used as resources for the production of PRR.

PhilRice Genebank: recent developments in managing and sharing the Philippine rice germplasm

The Genetic Resources Division (GRD) of PhilRice collects and conserves rice genetic resources to ensure the future generations of available seeds needed to build better rice plants in facing climate change and growing population. At present, GRD maintains the national collection of rice genetic resources with 7,129 accessions. To effectively manage the germplasm collection, the search for, development of, and implementation of the best conservation strategies and innovation in technology have been the utmost priority of the GRD. Thus, georeference data such as latitude, longitude and elevation of germplasm origin during collecting mission were recorded using a handheld global positioning system (GPS) receiver. The e-Seedfile software was developed to provide virtual access of the reference collection for regenerated germplasm seed verification and valid type confirmation for new and old germplasm collection. Barcoding, on the other hand, facilitated accurate inventory of seed stocks, making the distribution and regeneration of germplasm more efficient. Moreover, paperless data collection using android application was implemented for immediate data validation and accurate data downloading from tablets to workstations, making it an ideal tool for germplasm characterization. Furthermore, the current database system was upgraded and adjusted to adopt the use of digital object identifier (DOI) through registration to the global information system (GLIS) on Plant Genetic Resources for Food and Agriculture (PGRFA). The DOI allows the use of material to be tracked, thus meeting the legal obligations of the SMTA and monitor the impact of genebank collections in utilization in research and breeding programs. These innovative technologies are of great importance to expand the toolbox for the management and conservation of the germplasm collection that will help enhance the long-term conservation of rice diversity and easy access to germplasm and germplasm-related information.

Expected values and predictive model for black seed (Plaseolus vulgaris L.) production

The objective of this work was to determine the tendencies of expected values, linear associations, and to define which traits compose the best predictive model for mass of seeds per plant of black and pinto bean genotypes from different regions of Rio Grande do Sul. The experiments were conducted in the agricultural years of (2015-2017). The expected values are increased as function of the segregating generation of F2, F3, F4 for first pod insertion height and plant height. Furthermore, black commercial group determines superiority for number of seeds per plant, while pinto genotypes are closely related to increments of mass of seeds per plant. Linear associations are specific to germplasm origin, commercial group and segregating generations of bean genotypes regarding traits of agronomic interest. The predictive model created to increase mass of seeds per plant is specific for germplasm origins and commercial group of beans. Seed yield of pinto genotypes is maximized by plant height and number of seeds per plant. For genotypes of black commercial group, this increase is achieved through plant height, number of pods and seeds per plant in F3 and F4 segregating generations.

Cultivated Olive Diversification at Local and Regional Scales: Evidence From the Genetic Characterization of French Genetic Resources.

Molecular characterization of crop genetic resources is a powerful approach to elucidate the origin of varieties and facilitate local cultivar management. Here we aimed to decipher the origin and diversification of French local olive germplasm. The 113 olive accessions of the ex situ collection of Porquerolles were characterized with 20 nuclear microsatellites plus their plastid haplotype. We then compared this collection to Mediterranean olive varieties from the Worldwide Olive Germplasm Bank of Marrakech, Morocco. High genetic diversity was observed within local French varieties, indicating a high admixture level, with an almost equal contribution from the three main Mediterranean gene pools. Nearly identical and closely related genotypes were observed among French and Italian/Spanish varieties. A high number of parent–offspring relationships were also detected among French varieties and between French and two Italian varieties (‘Frantoio’ and ‘Moraiolo’) and the Spanish variety (‘Gordal Sevillana’). Our investigations indicated that French olive germplasm resulted from the diffusion of material from multiple origins followed by diversification based on parentage relationships between varieties. We strongly suggest that farmers have been actively selecting olives based on local French varieties. French olive agroecosystems more affected by unexpected frosts than southernmost regions could also be seen as incubators and as a bridge between Italy and Spain that has enhanced varietal olive diversification.

Natural Variation in Fatty Acid Composition of Diverse World Soybean Germplasms Grown in China

Soybean (Glycine max L. Merr.) is one of the most important crops in the world. Its major content of vegetable oil made it widely used for human consumption and several food industries. To investigate the variation in seed fatty acid composition of soybeans from different origins, a set of 633 soybean accessions originated from four diverse germplasm collections—including China, United States of America (USA), Japan, and Russia—were grown in three locations, Beijing, Anhui, and Hainan for two years. The results showed significant differences (P &lt; 0.001) among the four germplasm origins for all fatty acid contents investigated. Higher levels, on average, of palmitic acid (PA) and linolenic acid (LNA) were observed in Russian germplasm (12.31% and 8.15%, respectively), whereas higher levels of stearic acid (SA) and oleic acid (OA) were observed in Chinese germplasm (3.95% and 21.95%, respectively). The highest level of linoleic acid (LA) was noticed in the USA germplasm accessions (56.34%). The largest variation in fatty acid composition was found in LNA, while a large variation was observed between Chinese and USA germplasms for LA level. Maturity group (MG) significantly (P &lt; 0.0001) affected all fatty acids and higher levels of PA, SA, and OA were observed in early maturing accessions, while higher levels of LA and LNA were observed in late maturing accessions. The trends of fatty acids concentrations with different MG in this study further provide an evidence of the importance of MG in breeding for such soybean seed components. Collectively, the unique accessions identified in this study can be used to strengthen the soybean breeding programs for meeting various human nutrition patterns around the globe.

Genetic Parameters in the Initial Phase of Collaborative Plant Breeding in Lima-Beans (Phaseolus lunatus)

Participatory genetic improvement is a component of the management of genetic diversity, aiming at the systematic inclusion of farmers' knowledge, skills, experiences, practices and preferences. This modality of improvement is based on the knowledge of conventional genetics, physiology and economics, combined with those of anthropology, sociology, knowledge of the producers and with the principles of product development. The development of participatory strategies in the improvement of different crop species in the farmers field involves some fundamental aspects such as: rescue of different species and of different varieties within each species, valorization of local varieties, development of new varieties, valorization of cultural aspects and nutrition, agroecosystem management, agro-ecological crop systems, adaptation to the local environment with increased production from participatory improvement. Initially a search was made for communities in the forest area of ​​Pernambuco, Brazil, which preserve, store and use native varieties. After the identification of the localities contemplated. The seeds were obtained in the municipality of Lagoa de Itaenga. The proposal was submitted to farmers living in the Marrecos community. At the same time, the importance of the conservation of local cultivars, the maintenance of the genetic identity of these genotypes, the importance of the isolation of the cultures, mainly the Creole ones and the practice of the participatory improvement were also treated. In addition, seed samples were collected and stored by the farmers, for later implantation of the germplasm bank (in vivo) at the Federal Institute of Pernambuco. At the time of collection, the forms of storage adopted by farmers were observed, being of great relevance for the guarantee of the genetic reserves of the crops. The experiments were conducted in a phyto-bacteriology laboratory and in a greenhouse of the Department of Agriculture of the Federal Institute of Pernambuco (IFPE) in Vitória de Santo Antão-PE. The plants were cultivated in the field in an organic cultivation system. The results showed that there is genetic divergence, based on agro-morphological markers, among fava varieties studied as a function of germplasm origin. Thus, samples from the Germplasm Collection of IFPE-Camps Vitoria correspond to different materials. The existence of genetic divergence among the varieties indicates the possibility of selection of superior genotypes. The evaluated characteristics present high potential to differentiate the genotypes and should serve as parameters for other studies.

Genetic Diversity in broccoli rabe (Brassica rapa L. subsp. sylvestris (L.) Janch.) from Southern Italy

Broccoli rabe (Brassica rapa L. subsp. sylvestris (L.) Janch.) is a vegetable crop cultivated in southern Italy where it is part of local traditional recipes and heritage. Despite its economic importance, there are no cultivars patented, giving rise to many cultivated landraces that are characterized by wide phenotypic variability and named according to their harvesting time or the size and shape of their primary inflorescence. We report the first study of genetic diversity of 67 landraces of broccoli rabe collected in Apulia region and genotyped by means of 18 very informative SSR markers. A total of 243 alleles were detected in 552 genotypes, with a mean of 13.5 alleles per locus. The molecular variance was partitioned for 64% within accessions and for 36% among accessions, revealing a high level of genetic variation within landraces and a significant genetic differentiation across themselves. Remarkably, the results of genetic structure inference, as well as principal coordinates analysis and hierarchical clustering indicated as the clustering was largely in accordance to the germplasm origin, highlighting the presence of geographic stratification. We established a comprehensive germplasm collection of Apulian broccoli rabe landraces, currently stored at the Genebanks of IBBR-CNR and University of Bari. Finally, our study lays a foundation for future bio-agronomic characterizations addressing the identification of superior accessions for breeding purposes and solving cases of homonymy and synonymy.

Complete mitochondrial genome sequence of Zhuxiang chicken (Gallus gallus. domesticus) and its phylogenetic analysis from D-loop region

Zhuxiang chicken is a valuable chicken breed in the southwest of China. In this study, we firstly obtained the complete mitochondrial genome sequence of Zhuxiang chicken using PCR amplification, sequencing and assembling. The total length of the mitochondrial genome was 16,789 bp, based on composition of 30.24% for A, 23.71% for T, 32.54% for C, 13.51% for G, contained a D-loop region (non-coding control region), 2 ribosomal RNA genes, 13 protein-coding genes and 22 transfer RNA. This research was significant for mtDNA structure and provided phylogenetic analysis of D-loop sequence for carrying out Zhuxiang chicken germplasm resources protection, rational breeding, origin and evolution studies.

Identification and characterization of karyotype in Passiflora hybrids using FISH and GISH

BackgroundA great interest exists in the production of hybrid plants of the genus Passiflora given the beauty and exotic features of its flowers which have ornamental value. Hybrid paternity confirmation is therefore important for assuring germplasm origin, and is typically carried out by molecular marker segregation. The aim of this study was to karyotypically characterize the chromosome heritance patterns of the progeny resultant from a cross of P. gardneri and P. gibertii using classical cytogenetics, chromosome banding, and molecular cytogenetics.ResultsAll analyzed genotypes showed the same diploid chromosome number as the genitor species: 2n = 18. Classical and CMA3 and DAPI staining allowed for chromosome counting and satellite identification (secondary constrictions). Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used to characterize subgenomes by either identifying rDNA-specific genome patterns or parental genomes, respectively.ConclusionsThe heritance of chromosomal markers presenting rDNA sites from each parent for genome identification confirmed that all obtained plants were hybrids. These results will improve breeding programs involving the species of this genus. Apart from confirming hybridization, GISH allowed the visualization of recombination between the homeologous chromosome and the introgression of sequences of interest.