Pod indehiscence is a domestication and aridity resilience trait in common bean.

Plant domestication has strongly modified crop morphology and development. Nevertheless, many crops continue to display atavistic characteristics that were advantageous to their wild ancestors but are deleterious under cultivation, such as pod dehiscence (PD). Here, we provide the first comprehensive assessment of the inheritance of PD in the common bean (Phaseolus vulgaris), a major domesticated grain legume. Using three methods to evaluate the PD phenotype, we identified multiple, unlinked genetic regions controlling PD in a biparental population and two diversity panels. Subsequently, we assessed patterns of orthology among these loci and those controlling the trait in other species. Our results show that different genes were selected in each domestication and ecogeographic race. A chromosome Pv03 dirigent-like gene, involved in lignin biosynthesis, showed a base-pair substitution that is associated with decreased PD. This haplotype may underlie the expansion of Mesoamerican domesticates into northern Mexico, where arid conditions promote PD. The rise in frequency of the decreased-PD haplotype may be a consequence of the markedly different fitness landscape imposed by domestication. Environmental dependency and genetic redundancy can explain the maintenance of atavistic traits under domestication

Transcriptomic Studies in Non-Model Plants: Case of Pisum sativum L. and Medicago lupulina L.

Transcriptomics is a dynamically developing branch of biology highly important for geneticists and molecular ecologists alike. A large number of studies concerning differential gene expression, mapping of genes and quantitative trait loci (QTL), analysis of genotyping variations and so on has been conducted recently on several non‐model plants using next‐generation sequencing techniques. One example of non‐model legumes is garden pea (Pisum sativum L.), a valuable pulse crop capable of forming nitrogen‐fixing nodules and arbuscular mycorrhiza. Adaptation of standardised RNA‐seq approaches and data analysis developed for model plants to P. sativum should facilitate both studying of pea molecular genetics and breeding of new cultivars possessing agriculturally important traits. Another non‐model legume is black medick Medicago lupulina L. (a close relative of model legume plant barrel medick, Medicago truncatula Gaertn.), for which unique genetic lines almost obligatory dependent on arbuscular mycorrhiza symbiosis formation have been obtained. Such lines show promise as the perfect model for studying the genetic bases of arbuscular mycorrhiza development. In this chapter, we give a brief description of the current developments in the field of garden pea and black medick transcriptomics. Our aim is to provide a quick start guide to the non‐expert researchers for next‐generation sequencing (NGS)‐based transcriptome analysis

Genomic and phenotypic analysis of Vavilov's historic landraces reveals the impact of environment and genomic islands of agronomic traits.

The Vavilov Institute of Plant Genetic Resources (VIR), in St. Petersburg, Russia, houses a unique genebank, with historical collections of landraces. When they were collected, the geographical distribution and genetic diversity of most crops closely reflected their historical patterns of cultivation established over the preceding millennia. We employed a combination of genomics, computational biology and phenotyping to characterize VIR's 147 chickpea accessions from Turkey and Ethiopia, representing chickpea's center of origin and a major location of secondary diversity. Genotyping by sequencing identified 14,059 segregating polymorphisms and genome-wide association studies revealed 28 GWAS hits in potential candidate genes likely to affect traits of agricultural importance. The proportion of polymorphisms shared among accessions is a strong predictor of phenotypic resemblance, and of environmental similarity between historical sampling sites. We found that 20 out of 28 polymorphisms, associated with multiple traits, including days to maturity, plant phenology, and yield-related traits such as pod number, localized to chromosome 4. We hypothesize that selection and introgression via inadvertent hybridization between more and less advanced morphotypes might have resulted in agricultural improvement genes being aggregated to genomic 'agro islands', and in genotype-to-phenotype relationships resembling widespread pleiotropy

Genome-wide investigation and expression analysis of AP2-ERF gene family in salt tolerant common bean

Apetala2-ethylene-responsive element binding factor (AP2-ERF) superfamily with common AP2-DNA binding domain have developmentally and physiologically important roles in plants. Since common bean genome project has been completed recently, it is possible to identify all of the AP2-ERF genes in the common bean genome. In this study, a comprehensive genome-wide in silico analysis identified 180 AP2-ERF superfamily genes in common bean (Phaseolus vulgaris). Based on the amino acid alignment and phylogenetic analyses, superfamily members were classified into four subfamilies: DREB (54), ERF (95), AP2 (27) and RAV (3), as well as one soloist. The physical and chemical characteristics of amino acids, interaction between AP2-ERF proteins, cis elements of promoter region of AP2-ERF genes and phylogenetic trees were predicted and analyzed. Additionally, expression levels of AP2-ERF genes were evaluated by in silico and qRT-PCR analyses. In silico micro-RNA target transcript analyses identified nearly all PvAP2-ERF genes as targets of by 44 different plant species’ miRNAs were identified in this study. The most abundant target genes were PvAP2/ERF-20-25-62-78-113-173. miR156, miR172 and miR838 were the most important miRNAs found in targeting and BLAST analyses. Interactome analysis revealed that the transcription factor PvAP2-ERF78, an ortholog of Arabidopsis At2G28550, was potentially interacted with at least 15 proteins, indicating that it was very important in transcriptional regulation. Here we present the first study to identify and characterize the AP2-ERF transcription factors in common bean using whole-genome analysis, and the findings may serve as a references for future functional research on the transcription factors in common bean

Genomic resources in plant breeding for sustainable agriculture

Climate change during the last 40 years has had a serious impact on agriculture and threatens global food and nutritional security. From over half a million plant species, cereals and legumes are the most important for food and nutritional security. Although systematic plant breeding has a relatively short history, conventional breeding coupled with advances in technology and crop management strategies has increased crop yields by 56 % globally between 1965-85, referred to as the Green Revolution. Nevertheless, increased demand for food, feed, fiber, and fuel necessitates the need to break existing yield barriers in many crop plants. In the first decade of the 21st century we witnessed rapid discovery, transformative technological development and declining costs of genomics technologies. In the second decade, the field turned towards making sense of the vast amount of genomic information and subsequently moved towards accurately predicting gene-to-phenotype associations and tailoring plants for climate resilience and global food security. In this review we focus on genomic resources, genome and germplasm sequencing, sequencing-based trait mapping, and genomics-assisted breeding approaches aimed at developing biotic stress resistant, abiotic stress tolerant and high nutrition varieties in six major cereals (rice, maize, wheat, barley, sorghum and pearl millet), and six major legumes (soybean, groundnut, cowpea, common bean, chickpea and pigeonpea). We further provide a perspective and way forward to use genomic breeding approaches including marker-assisted selection, marker-assisted backcrossing, haplotype based breeding and genomic prediction approaches coupled with machine learning and artificial intelligence, to speed breeding approaches. The overall goal is to accelerate genetic gains and deliver climate resilient and high nutrition crop varieties for sustainable agriculture

Comparative analysis of plant genomes through data integration

When we started our research in 2008, several online resources for genomics existed, each with a different focus. TAIR (The Arabidopsis Information Resource) has a focus on the plant model species Arabidopsis thaliana, with (at that time) little or no support for evolutionary or comparative genomics. Ensemble provided some basic tools and functions as a data warehouse, but it would only start incorporating plant genomes in 2010. There was no online resource at that time however, that provided the necessary data content and tools for plant comparative and evolutionary genomics that we required. As such, the plant community was missing an essential component to get their research at the same level as the biomedicine oriented research communities. We started to work on PLAZA in order to provide such a data resource that could be accessed by the plant community, and which also contained the necessary data content to help our research group’s focus on evolutionary genomics. The platform for comparative and evolutionary genomics, which we named PLAZA, was developed from scratch (i.e. not based on an existing database scheme, such as Ensemble). Gathering the data for all species, parsing this data into a common format and then uploading it into the database was the next step. We developed a processing pipeline, based on sequence similarity measurements, to group genes into gene families and sub families. Functional annotation was gathered through both the original data providers and through InterPro scans, combined with Interpro2GO. This primary data information was then ready to be used in every subsequent analysis. Building such a database was good enough for research within our bioinformatics group, but the target goal was to provide a comprehensive resource for all plant biologists with an interest in comparative and evolutionary genomics. Designing and creating a user-friendly, visually appealing web interface, connected to our database, was the next step. While the most detailed information is commonly presented in data tables, aesthetically pleasing graphics, images and charts are often used to visualize trends, general statistics and also used in specific tools. Design and development of these tools and visualizations is thus one of the core elements within my PhD. The PLAZA platform was designed as a gene-centric data resource, which is easily navigated when a biologist wants to study a relative small number of genes. However, using the default PLAZA website to retrieve information for dozens of genes quickly becomes very tedious. Therefore a ’gene set’-centric extra layer was developed where user-defined gene sets could be quickly analyzed. This extra layer, called the PLAZA workbench, functions on top of the normal PLAZA website, implicating that only gene sets from species present within the PLAZA database can be directly analyzed. The PLAZA resource for comparative and evolutionary genomics was a major success, but it still had several issues. We tried to solve at least two of these problems at the same time by creating a new platform. The first issue was the building procedure of PLAZA: adding a single species, or updating the structural annotation of an existing one, requires the total re-computation of the database content. The second issue was the restrictiveness of the PLAZA workbench: through a mapping procedure gene sets could be entered for species not present in the PLAZA database, but for species without a phylogenetic close relative this approach did not always yield satisfying results. Furthermore, the research in question might just focus on the difference between a species present in PLAZA and a close relative not present in PLAZA (e.g. to study adaptation to a different ecological niche). In such a case, the mapping procedure is in itself useless. With the advent of NGS transcriptome data sets for a growing number of species, it was clear that a next challenge had presented itself. We designed and developed a new platform, named TRAPID, which could automatically process entire transcriptome data sets, using a reference database. The target goal was to have the processing done quickly with the results containing both gene family oriented data (such as multiple sequence alignments and phylogenetic trees) and functional characterization of the transcripts. Major efforts went into designing the processing pipeline so it could be reliable, fast and accurate

Specific tissue proteins 1 and 6 are involved in root biology during normal development and under symbiotic and pathogenic interactions in Medicago truncatula

Specifc tissue (ST) proteins have been shown to be involved in several processes related to plant nutritional status, development, and responses to biotic agents. In particular, ST1 and ST6 are mainly expressed in roots throughout plant development. Here, we analyze where and how the expression of the genes encoding both proteins are modulated in the legume model plant Medicago truncatula in response to the plant developmental program, nodulation induced by a benefcial nitrogen-fxing bacterium (Sinorhizobium meliloti) and the defense response triggered by a pathogenic hemibiotrophic fungus (Fusarium oxysporum). Gene expression results show that ST1 and ST6 participate in the vasculature development of both primary and lateral roots, although only ST6 is related to meristem activity. ST1 and ST6 clearly display diferent roles in the biotic interactions analyzed, where ST1 is activated in response to a N2-fxing bacterium and ST6 is up-regulated after inoculation with F. oxysporum. The role of ST1 and ST6 in the nodulation process may be related to nodule organogenesis rather than to the establishment of the interaction itself, and an increase in ST6 correlates with the activation of the salicylic acid signaling pathway during the infection and colonization processes. These results further support the role of ST6 in response to hemibiotrophic fungi. This research contributes to the understanding of the complex network that controls root biology and strengthens the idea that ST proteins are involved in several processes such as primary and lateral root development, nodule organogenesis, and the plant–microbe interaction

The Kirkhouse Trust: Successes and Challenges in Twenty Years of Supporting Independent, Contemporary Grain Legume Breeding Projects in India and African Countries

This manuscript reviews two decades of projects funded by the Kirkhouse Trust (KT), a charity registered in the UK. KT was established to improve the productivity of legume crops important in African countries and in India. KT’s requirements for support are: (1) the research must be conducted by national scientists in their home institution, either a publicly funded agricultural research institute or a university; (2) the projects need to include a molecular biology component, which to date has mostly comprised the use of molecular markers for the selection of one or more target traits in a crop improvement programme; (3) the projects funded are included in consortia, to foster the creation of scientific communities and the sharing of knowledge and breeding resources. This account relates to the key achievements and challenges, reflects on the lessons learned and outlines future research priorities

Study of inheritance and identification of molecular markers for seed protein content in pigeonpea (Cajanus cajan (L.) Millsp.).

Doctor of Philosophy in Plant Breeding. University of KwaZulu-Natal, Pietermaritzburg 2017.Abstract available in PDF file

Characterization of transport proteins on the symbiosome membrane of Glycine max

During the legume-rhizobia symbiosis, a new plant organ called the root nodule is induced and developed. Within the root nodule, rhizobia are enclosed in the symbiosome. The symbiosome is an organelle where rhizobia fix atmospheric nitrogen and is surrounded by the plant-derived symbiosome membrane (SM). The SM regulates nutrient exchange between the symbionts, and thus effectively controls the symbiosis. GmABCA1 and GmABCA2 proteins have been identified by proteomic analysis on the SM of soybean. These proteins are part of the large ATP-binding cassette (ABC) superfamily that transports diverse substrates and plays essential roles in plant growth and development. The present work aims to characterize GmABCA1 and GmABCA2, and to explore potential function they may play in relation to fatty acid transport. Expression analyses showed that they are preferentially expressed in the nodule tissue, and have specific expression in infected cells. The localization of GmABCA2 on the SM was confirmed by FP tagging but that of GmABCA1 could not be determined by this method. Complementation of the yeast fatty acid uptake-deficient mutant, Δfat1, indicated that both proteins were able to transport oleic acid. They also complemented the T-DNA mutant for the Arabidopsis fatty acid transporter Atabca9. The nitrate transporter 1/peptide transporter family (NPF) is mainly involved in the transport of nitrates and peptides in plants. Complementation of eight soybean NPF members with enhanced expression in nodule: GmNPF1.2, GmNPF5.24, GmNPF5.25, GmNPF5.29, GmNPF5.30, GmNPF5.2, GmNPF5.3 and GmNPF8.6 in a yeast peptide transport mutant, ptr2, was attempted in this study. Only GmNPF8.6 was able to restore ptr2 yeast growth on minimal media containing tri-alanine, glycine-proline and valine-leucine peptides as the sole source of nitrogen. This study provides the first evidence of such transport across the SM