Genomic analysis of a spontaneous unifoliate mutant reveals gene candidates associated with compound leaf development in Vigna unguiculata [L] Walp.

Molecular mechanisms which underpin compound leaf development in some legumes have been reported, but there is no previous study on the molecular genetic control of compound leaf formation in Vigna unguiculata (cowpea), an important dryland legume of African origin. In most studied species with compound leaves, class 1 KNOTTED-LIKE HOMEOBOX genes expressed in developing leaf primordia sustain morphogenetic activity, allowing leaf dissection and the development of leaflets. Other genes, such as, SINGLE LEAFLET1 in Medicago truncatula and Trifoliate in Solanum lycopersicum, are also implicated in regulating compound leaf patterning. To set the pace for an in-depth understanding of the genetics of compound leaf development in cowpea, we applied RNA-seq and whole genome shotgun sequence datasets of a spontaneous cowpea unifoliate mutant and its trifoliate wild-type cultivar to conduct comparative reference-based gene expression, de novo genome-wide isoform switch, and genome variant analyses between the two genotypes. Our results suggest that genomic variants upstream of LATE ELONGATED HYPOCOTYL and down-stream of REVEILLE4, BRASSINOSTERIOD INSENSITIVE1 and LATERAL ORGAN BOUNDARIES result in down-regulation of key components of cowpea circadian rhythm central oscillator and brassinosteroid signaling, resulting in unifoliate leaves and brassinosteroid-deficient-like phenotypes. We have stated hypotheses that will guide follow-up studies expected to provide more insights.

Cowpea speed breeding using regulated growth chamber conditions and seeds of oven-dried immature pods potentially accommodates eight generations per year.

BACKGROUND: Cowpea is a dryland crop with potential to improve food security in sub-Saharan Africa, where it is mostly produced and consumed. Contemporary plant improvement technologies, including genome editing, marker-assisted selection, and optimized transformation protocols, are being deployed to improve cowpea characteristics. Integrating speed breeding with these technologies would accelerate genetic gain in cowpea breeding. There are established speed breeding protocols for other important legumes, such as soybean, peanut, and chickpea, but none has been previously reported for cowpea. RESULTS: With the aid of regulated growth conditions in two different chamber types, as well as the cultivation of new plant generations from seeds of oven-dried immature pods, we developed and validated, for the first time, an efficient speed breeding protocol that accommodates approximately seven to eight breeding generations per year for 3 cowpea genotypes. The 3 cowpea genotypes were evaluated under controlled growth conditions in light-emitting diode and metal halide lamp chambers to determine the effect of CO2 supplementation on flowering and maturation durations, optimum conditions for plant growth, cross pollination, and pod development. Elevated CO2 concentration had no influence on either flowering time or pod development. Adequate temperature, relative humidity and light intensity improved plant development and the rate of successful hand pollination, and  cultivating seeds of 11-day-old immature pods oven-dried at 39 °C for 2 days resulted in at least a 62% reduction in the time between pollination and sowing of the next plant generation. The plants cultivated from seeds of the oven-dried immature pods showed no defect at any stage of development. CONCLUSIONS: Using the speed breeding protocol developed in this study, cowpea breeding cycles can be increased from the traditional one cycle per year in the field to as many as 8 generations per year in regulated growth chamber conditions. This protocol has no special technical requirements; hence, it can be implemented in any standard growth chamber. This would fast-track development, testing, validation, and utilization of improved cowpea cultivars.

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

BACKGROUND: The 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. RESULTS: We 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. CONCLUSION: The 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.