Genetic Characterization and Curation of Diploid A-Genome Wheat Species

The A-genome diploid wheats represent the earliest domesticated and cultivated wheat species in the Fertile Crescent and the donor of the wheat A sub-genome. The A-genome species encompass the cultivated einkorn (Triticum. monococcum L. subsp. monococcum), wild einkorn (T. monococcum L. subsp. aegilopoides (Link) Thell.) and T. urartu. We evaluated the collection of 930 accessions in the Wheat Genetics Resource Center (WGRC), using genotyping-by-sequencing (GBS) and identified 13,089 curated SNPs. Genomic analysis detected misclassified and duplicated accessions with most duplicates originated from the same or a nearby locations. About 56% (n = 520) of the WGRC A-genome species collections were duplicates supporting the need for genomic characterization for effective curation and maintenance of these collections. Population structure analysis confirmed the morphology-based classifications of the accessions and reflected the species geographic distributions. We also showed that the T. urartu as the closest A-genome diploid to wheat through phylogenetic analysis. Population analysis within the wild einkorn group showed three genetically distinct clusters, which corresponded with wild einkorn races α, β, and γ described previously. The T. monococcum genome-wide FST scan identified candidate genomic regions harboring domestication selection signature (Btr1) on the short arm of chromosome 3Am at ~ 70 Mb. We established A-genome core set (79 accessions) based on allelic diversity, geographical distribution, and available phenotypic data. The individual species core set maintained at least 80% of allelic variants in the A-genome collection and constitute a valuable genetic resource to improve wheat and domesticated einkorn in breeding programs.

Arachis inflata: A New B Genome species of Arachis (Fabaceae)

Great efforts have been done to collect germplasm of the Arachis genus in South America, however, many regions still remain underexplored. Under the hypothesis that these regions have new and diverse populations/species of Arachis, several expeditions were carried out since 2000 in Bolivia, to increase the documentation of the genus diversity. As a first result of these explorations, a new species of section Arachis with B genome is formally described. Arachis inflata is closely related to A. magna and A. ipaënsis, but it can be clearly distinguished from them, and from any other species of the genus, for having a type of fruit with a completely distinct morphology. The fruit has a smooth epicarp, but shows a bullated aspect, due to the presence of air chambers in the mesocarp.

Development of a genome-wide InDel marker set for allele discrimination between rice (Oryza sativa) and the other seven AA-genome Oryza species

Wild relatives of rice in the genus Oryza (composed of 24 species with 11 different genome types) have been significantly contributing to the varietal improvement of rice (Oryza sativa). More than 4000 accessions of wild rice species are available and they are regarded as a “genetic reservoir” for further rice improvement. DNA markers are essential tools in genetic analysis and breeding. To date, genome-wide marker sets for wild rice species have not been well established and this is one of the major difficulties for the efficient use of wild germplasm. Here, we developed 541 genome-wide InDel markers for the discrimination of alleles between the cultivated species O. sativa and the other seven AA-genome species by positional multiple sequence alignments among five AA-genome species with four rice varieties. The newly developed markers were tested by PCR-agarose gel analysis of 24 accessions from eight AA genome species (three accessions per species) along with two representative cultivars (O. sativa subsp. indica cv. IR24 and subsp. japonica cv. Nipponbare). Marker polymorphism was validated for 475 markers. The number of polymorphic markers between IR24 and each species (three accessions) ranged from 338 (versus O. rufipogon) to 416 (versus O. longistaminata) and the values in comparison with Nipponbare ranged from 179 (versus O. glaberrima) to 323 (versus O. glumaepatula). These marker sets will be useful for genetic studies and use of the AA-genome wild rice species.

Exploring the Loci Responsible for Awn Development in Rice through Comparative Analysis of All AA Genome Species

Wild rice species have long awns at their seed tips, but this trait has been lost through rice domestication. Awn loss mitigates harvest and seed storage; further, awnlessness increases the grain number and, subsequently, improves grain yield in Asian cultivated rice, highlighting the contribution of the loss of awn to modern rice agriculture. Therefore, identifying the genes regulating awn development would facilitate the elucidation of a part of the domestication process in rice and increase our understanding of the complex mechanism in awn morphogenesis. To identify the novel loci regulating awn development and understand the conservation of genes in other wild rice relatives belonging to the AA genome group, we analyzed the chromosome segment substitution lines (CSSL). In this study, we compared a number of CSSL sets derived by crossing wild rice species in the AA genome group with the cultivated species Oryza sativa ssp. japonica. Two loci on chromosomes 7 and 11 were newly discovered to be responsible for awn development. We also found wild relatives that were used as donor parents of the CSSLs carrying the functional alleles responsible for awn elongation, REGULATOR OF AWN ELONGATION 1 (RAE1) and RAE2. To understand the conserveness of RAE1 and RAE2 in wild rice relatives, we analyzed RAE1 and RAE2 sequences of 175 accessions among diverse AA genome species retrieved from the sequence read archive (SRA) database. Comparative sequence analysis demonstrated that most wild rice AA genome species maintained functional RAE1 and RAE2, whereas most Asian rice cultivars have lost either or both functions. In addition, some different loss-of-function alleles of RAE1 and RAE2 were found in Asian cultivated species. These findings suggest that different combinations of dysfunctional alleles of RAE1 and RAE2 were selected after the speciation of O. sativa, and that two-step loss of function in RAE1 and RAE2 contributed to awnlessness in Asian cultivated rice.

A Genetic Resource for Rice Improvement：introgression Library of Agronomic Traits for All AA Genome Species in Genus Oryza

Background: Rice improvement depends on the availability of genetic variation, and AA genome Oryza species are the natural reservoir of favorable genes which are useful for rice breeding. Developing the introgression library using multiple AA genome species was rarely reported.Results: In this study, to systematically evaluate and utilize potentially valuable QTLs/genes or allelic variations, based on the evaluation and selection of agronomic traits, 6372 introgression lines (ILs) were raised by crossing 330 accessions of 7 AA genome species as the donor parents, with three elite cultivars of O. sativa, Dianjingyou 1, Yundao 1 and RD23 as the recurrent parents, respectively. Further, twenty-six, twenty-six and nineteen loci were detected in the multiple donors using 1,401 ILs in the Dianjingyou 1 background for grain length, grain width, and the ratio of grain length to grain width, respectively. Interestingly, ten loci had opposite effect on grain length in the different donors, so did grain width. Moreover, one locus for grain width, qGW3.1, was validated using the segregation population derived from the donor of O. glumaepatula. Conclusions: This introgression library provided the powerful resource for future rice improvement and genetic dissection of allelic variations. Selections of favorable alleles that are present in wild relatives proceed the driving force of the rice domestication.

INDUKSI TUNAS PADA BEBERAPA TIPE PEMOTONGAN EKSPLAN BONGGOL PISANG UDANG (Musa acuminata Colla) SECARA IN VITRO

Musa acuminata Colla is a unique genome species different from other species. This species has morphological characteristics on its reddish-purple fruit colour. Nowadays, Musa acuminata Colla is rarely to fond in Riau especially in Kampar District. This research aims to know the influence of several kinds of corm bananas cutting that comes from Kampar district with in vitro and determined concentration of single BAP and combination of BAP and Kinetin best in forming Musa acuminata Colla shoot. This research used for randomized block design (RBD) by giving BAP concentration (0, 4, 8 mg/l and combination of BAP and Kinetin (0 mg/l BAP+0,4 mg/l Kinetin, 4 mg/l BAP+0,4 mg/l Kinetin, 8 mg/l BAP+ 0,4 mg/l Kinetin) with different cutting type the whole and cut into two parts on MS media with 5 replications. The result of this research showed the addition of BAP dan combination of BAP and Kinetin is giving the best result on the percentage of live explants and shot formation at 100%. Treatment of 8 mg/l BAP resulted in the highest shoot percentage up to 100%, the fastest shoots appeared at 34.00 days after planting and the highest shoot length of 2.83 cm with the shoots of 2.33 on cutting halved. Keywords : BAP, in vitro, kinetin, Musa acuminata Colla, shoot induction

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 Oryza officinalis, and draft genomes for two other C genome diploid species O. eichingeri and O. 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 O. sativa, mostly due to proliferation of Gypsy type long-terminal repeat (LTR) 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, O. eichingeri and O. 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 LTR 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.

Comparative Functional Studies on Two Diploid Cotton Genomes Reveals Functional Differences of Basic Helix-loop-helix Proteins in Arabidopsis Trichome Initiation

Background: The cultivated tetraploid cotton species (AD genomes) was originated from two ancestral diploid species (A and D genomes). While the ancestral A-genome species produce spinnable fibers, the D- genome species do not. Cotton fibers are unicellular trichomes originating from seed coat epidermal cells, and currently there is an immense interest in understanding the process of fiber initiation and development. Current knowledge demonstrates that there is a great of deal of resemblance in initiation mechanism between by Arabidopsis trichome and cotton fiber. Methodology: In this study, we performed comparative functional studies between A genome and D-genome species in cotton by using Arabidopsis trichome initiation as a model. Four cotton genes TTG3, MYB2, DEL61 and DEL65 were amplified from A-genome and D-genome species, and transformed into their homolog trichomeless mutants Arabidopsis ttg1, gl1, and gl3egl3, respectively. Results: Our data indicated that the transgenic plants expressing TTG3 and MYB2 genes from A-genome and D-genome species complement the ttg1 and gl1 mutants, respectively. We also discovered complete absences of two functional basic helix loop helix (bHLH) proteins (DEL65/DEL61) in D- diploid species and one (DEL65) that is functional in A-genome species, but not from D-genome species. This observation is consistent with the natural phenomenon of spinnable fiber production in A- genome species and absence in D-genome species.

Twelve complete chloroplast genomes of wild peanuts: great genetic resources and a better understanding of Arachis phylogeny

Background The cultivated peanut (Arachis hypogaea) is one of the most important oilseed crops worldwide, however, its improvement is restricted by its narrow genetic base. The highly variable wild peanut species, especially within Sect. Arachis, may serve as a rich genetic source of favorable alleles to peanut improvement; Sect. Arachis is the biggest taxonomic section within genus Arachis and its members also include the cultivated peanut. In order to make good use of these wild resources, the genetic bases and the relationships of the Arachis species need first to be better understood. Results Here, in this study, we have sequenced and/or assembled twelve Arachis complete chloroplast (cp) genomes (eleven from Sect. Arachis). These cp genome sequences enriched the published Arachis cp genome data. From the twelve acquired cp genomes, substantial genetic variation (1368 SNDs, 311 indels) has been identified, which, together with 69 SSR loci that have been identified from the same data set, will provide powerful tools for future explorations. Phylogenetic analyses in our study have grouped the Sect. Arachis species into two major lineages (I & II), this result together with reports from many earlier studies show that lineage II is dominated by AA genome species that are mostly perennial, while lineage I includes species that have more diverse genome types and are mostly annual/biennial. Moreover, the cultivated peanuts and A. monticola that are the only tetraploid (AABB) species within Arachis are nested within the AA genome species-dominated lineage, this result together with the maternal inheritance of chloroplast indicate a maternal origin of the two tetraploid species from an AA genome species. Conclusion In summary, we have acquired sequences of twelve complete Arachis cp genomes, which have not only helped us better understand how the cultivated peanut and its close wild relatives are related, but also provided us with rich genetic resources that may hold great potentials for future peanut breeding.