Domesticated strawberry is susceptible to sudden frost episodes, limiting the productivity of this cash crop in regions, where they are grown during early spring. In contrast, the ancestral woodland strawberry (Fragaria vesca) has successfully colonised many habitats of the Northern Hemisphere. Thus, this species seems to harbour genetic factors promoting cold tolerance. Screening a germplasm established in frame of the German Gene Bank for Crop Wild Relatives we identified, among 70 wild accessions, a pair contrasting with respect to cold tolerance. By following the physiological, biochemical, molecular, and metabolic responses of this contrasting pair, we identified the transcription factor Cold Box Factor 4 and the dehydrin Xero-2 as molecular markers associated with superior tolerance to cold stress. Overexpression of GFP fusions with Xero-2 in tobacco BY-2 cells conferred cold tolerance to these recipient cells. A detailed analysis of the metabolome for the two contrasting genotypes allows to define metabolic signatures correlated with cold tolerance versus cold stress. This work provides a proof-of-concept for the value of crop wild relatives as genetic resources to identify genetic factors suitable to increase the stress resilience of crop plants.

The genetic diversity found in natural populations is the result of the evolutionary forces in response to historical and contemporary factors. The environmental characteristics and geological history of Mexico promoted the evolution and diversification of plant species, including wild relatives of crops such as the wild pumpkins (Cucurbita). Wild pumpkin species are found in a variety of habitats, evidencing their capability to adapt to different environments. Despite the potential value of wild Cucurbita as a genetic reservoir for crops, there is a lack of studies on their genetic diversity. Cucurbita radicans is an endangered species threatened by habitat destruction leading to low densities in small and isolated populations. Here, we analyze Genotype by Sequencing genomic data of the wild pumpkin C. radicans to evaluate the influence of factors like isolation, demographic history, and the environment shaping the amount and distribution of its genetic variation. We analyzed 91 individuals from 14 localities along its reported distribution. We obtained 5,107 SNPs and found medium-high levels of genetic diversity and genetic structure distributed in four main geographic areas with different environmental conditions. Moreover, we found signals of demographic growth related to historical climatic shifts. Outlier loci analysis showed significant association with the environment, principally with precipitation variables. Also, the outlier loci displayed differential changes in their frequencies in response to future global climate change scenarios. Using the results of genetic structure, outlier loci and multivariate analyses of the environmental conditions, we propose priority localities for conservation that encompass most of the genetic diversity of C. radicans.

Chickpea (Cicer arietinum) is a major food legume providing high quality nutrition, especially in developing regions. Chickpea wilt (Fusarium oxysporum f. sp. ciceris) causes significant annual losses. Integrated disease management of Fusarium wilt is supported by resistant varieties. Relatively few resistance genes are known so there is value in exploring genetic resources in chickpea wild relatives. This study investigates the inheritance of Fusarium wilt resistance (race 2) in recombinant inbred lines (RILs) from a cross between a cultivated susceptible chickpea variety (Gokce) and a wild resistant Cicer reticulatum line (Kayat-077). RILs, parents, resistant and susceptible tester lines were twice grown in the greenhouse with inoculation and disease symptoms scored. DNA was extracted from dried leaves and individuals were single nucleotide polymorphism (SNP) genotyped. SNPs were placed on the reference chickpea genome and quantitative trait locus (QTL) mapping was performed. Significant QTL regions were examined using PulseDB to identify candidate genes. The results showed the segregation of Fusarium wilt resistance conforming to a single gene inheritance. One significant QTL was found at the start of chromosome 8, containing 138 genes, three of which were disease-resistance candidates for chickpea breeding.

Ensuring global food security in the face of climate change is critical to human survival. With a predicted human population of 9.6 billion in 2050 and the demand for food supplies expected to increase by 60% globally, but with a parallel potential reduction in crop production for wheat by 6.0%, rice by 3.2%, maize by 7.4%, and soybean by 3.1% by the end of the century, maintaining future food security will be a challenge. One potential solution is new climate-smart varieties created using the breadth of diversity inherent in crop wild relatives (CWRs). Yet CWRs are threatened, with 16-35% regarded as threatened and a significantly higher percentage suffering genetic erosion. Additionally, they are under-conserved, 95% requiring additional ex situ collections and less than 1% being actively conserved in situ; they also often grow naturally in disturbed habitats limiting standard conservation measures. The urgent requirement for active CWR conservation is widely recognized in the global policy context (Convention on Biological Diversity post-2020 Global Biodiversity Framework, UN Sustainable Development Goals, the FAO Second Global Plan of Action for PGRFA, and the FAO Framework for Action on Biodiversity for Food and Agriculture) and breeders highlight that the lack of CWR diversity is unnecessarily limiting crop improvement. CWRs are not spread evenly across the globe; they are focused in hotspots and the hottest region for CWR diversity is in West Asia and North Africa (WANA). The region has about 40% of global priority taxa and the top 17 countries with maximum numbers of CWR taxa per unit area are all in WANA. Therefore, improved CWR active conservation in WANA is not only a regional but a critical global priority. To assist in the achievement of this goal, we will review the following topics for CWRs in the WANA region: (1) conservation status, (2) community-based conservation, (3) threat status, (4) diversity use, (5) CURE-CWR hub: (ICARDA Centre of Excellence), and (6) recommendations for research priorities. The implementation of the recommendations is likely to significantly improve CWRs in situ and ex situ conservation and will potentially at least double the availability of the full breadth of CWR diversity found in WANA to breeders, and so enhance regional and global food and nutritional security.

OBJECTIVE: The timing of flowering onset is often correlated with latitude, indicative of climatic gradients. Flowering onset in temperate species commonly requires exposure to cold temperatures, known as vernalization. Hence, population differentiation of flowering onset with latitude might reflect adaptation to the local climatic conditions experienced by populations.
METHODS: Within its western range, seeds from Linum bienne populations (the wild relative of cultivated Linum usitatissimum) were used to describe the latitudinal differentiation of flowering onset to determine its association with the local climate of the population. A vernalization experiment including different crop cultivars was used to determine how vernalization accelerates flowering onset, in addition to the vernalization sensitivity response among populations and cultivars. Additionally, genetic differentiation of L. bienne populations along the latitudinal range was scrutinized using microsatellite markers.
RESULTS: Flowering onset varied with latitude of origin, with southern populations flowering earlier than their northern counterparts. Vernalization reduced the number of days to flowering onset, but vernalization sensitivity was greater in northern populations compared with southern ones. Conversely, vernalization delayed flowering onset in the crop, exhibiting less variation in sensitivity. In L. bienne, both flowering onset and vernalization sensitivity were better predicted by the local climate of the population than by latitude itself. Microsatellite data unveiled genetic differentiation of populations, forming two groups geographically partitioned along latitude.
CONCLUSIONS: The consistent finding of latitudinal variation across experiments suggests that both flowering onset and vernalization sensitivity in L. bienne populations are under genetic regulation and might depend on climatic cues at the place of origin. The association with climatic gradients along latitude suggests that the climate experienced locally drives population differentiation of the flowering onset and vernalization sensitivity patterns. The genetic population structure suggests that past population history could have influenced the flowering initiation patterns detected, which deserves further work.

The Pacific crabapple (Malus fusca) is a wild relative of the commercial apple (Malus × domestica). With a range extending from Alaska to Northern California, M. fusca is extremely hardy and disease resistant. The species represents an untapped genetic resource for the development of new apple cultivars with enhanced stress resistance. However, gene discovery and utilization of M. fusca have been hampered by the lack of genomic resources. Here, we present a high-quality, haplotype-resolved, chromosome-scale genome assembly and annotation for M. fusca. The genome was assembled using high-fidelity long-reads and scaffolded using genetic maps and high-throughput chromatin conformation capture sequencing, resulting in one of the most contiguous apple genomes to date. We annotated the genome using public transcriptomic data from the same species taken from diverse plant structures and developmental stages. Using this assembly, we explored haplotypic structural variation within the genome of M. fusca, identifying thousands of large variants. We further showed high sequence co-linearity with other domesticated and wild Malus species. Finally, we resolve a known quantitative trait locus associated with resistance to fire blight (Erwinia amylovora). Insights gained from the assembly of a reference-quality genome of this hardy wild apple relative will be invaluable as a tool to facilitate DNA-informed introgression breeding.

Dasypyrum villosum is one of the most valuable gene resources in wheat improvement, especially for disease resistance. The mining of favorable genes from D. villosum is frustrated by the lack of a whole genome sequence. In this study, we generated a doubled-haploid line, 91C43DH, using microspore culture and obtained a 4.05-GB high-quality, chromosome-scale genome assembly for D. villosum. The assembly contains39 727 high-confidence genes, and 85.31% of the sequences are repetitive. Two reciprocal translocation events were detected, and 7VS-4VL is a unique translocation in D. villosum. The prolamin seed storage protein-coding genes were found to be duplicated; in particular, the genes encoding low-molecular-weight glutenin at the Glu-V3 locus were significantly expanded. RNA sequencing (RNA-seq) analysis indicated that, after Blumeria graminearum f.sp tritici (Bgt) inoculation, there were more upregulated genes involved in the pattern-triggered immunity and effector-triggered immunity defense pathways in D. villosum than in Triticum urartu. MNase hypersensitive sequencing (MH-seq) identified two Bgt-inducible MH sites (MHSs), one in the promoter and one in the 3\' terminal region of the powdery mildew resistance (Pm) gene NLR1-V. Each site had two subpeaks and they were termed MHS1 (MHS1.1/1.2) and MHS2 (MHS2.1/2.2). Bgt-inducible MHS2.2 was uniquely present in D. villosum, and MHS1.1 was more inducible in D. villosum than in wheat, suggesting that MHSs may be critical for regulation of NLR1-V expression and plant defense. In summary, this study provides a valuable genome resource for functional genomics studies and wheat-D. villosum introgression breeding. The identified regulatory mechanisms may also be exploited to develop new strategies for enhancing Pm resistance by optimizing gene expression in wheat.

Domestication is considered a model of adaptation that can be used to draw conclusions about the modus operandi of selection in natural systems. Investigating domestication may give insights into how plants react to different intensities of human manipulation, which has direct implication for the continuing efforts of crop improvement. Therefore, scientists of various disciplines study domestication-related questions to understand the biological and cultural bases of the domestication process. We employed restriction site-associated DNA sequencing (RAD-seq) of 494 Pisum sativum (pea) samples from all wild and domesticated groups to analyze the genetic structure of the collection. Patterns of ancient admixture were investigated by analysis of admixture graphs. We used two complementary approaches, one diversity based and one based on differentiation, to detect the selection signatures putatively associated with domestication. An analysis of the subpopulation structure of wild P. sativum revealed five distinct groups with a notable geographic pattern. Pisum abyssinicum clustered unequivocally within the P. sativum complex, without any indication of hybrid origin. We detected 32 genomic regions putatively subjected to selection: 29 in P. sativum ssp. sativum and three in P. abyssinicum. The two domesticated groups did not share regions under selection and did not display similar haplotype patterns within those regions. Wild P. sativum is structured into well-diverged subgroups. Although Pisum sativum ssp. humile is not supported as a taxonomic entity, the so-called \'southern humile\' is a genuine wild group. Introgression did not shape the variation observed within the sampled germplasm. The two domesticated pea groups display distinct genetic bases of domestication, suggesting two genetically independent domestication events.

The declining trends in crop wild relative genetic resources in many crop centers of origins including Ethiopia require short and long-term conservation strategies. Enset (Ensete ventricosum) is arguably the most important cultivated food security crop of Ethiopia with dwindling wild stocks. The cultivated enset is propagated clonally through adventitious bud sprouting from the corm after the distraction of the apical meristem. Shoot regeneration in the cultivated enset has been induced by humans and has not been observed to occur naturally. The technique of shoot induction has not been extended to the wild enset. To determine whether the capacity for shoot regeneration existed in wild enset and optimize the technique, a series of experiments were conducted. These involved: (i) sucker production from corms of wild enset with and without apical meristem removal; (ii) sprouting capacity of corms ranging 22-49 cm diameter, with removed apical meristem; and (iii) a factorial experiment involving two populations of wild enset (from Shebena and Getiba localities in Sheka zone), two ways of preparing or cutting the corms: tero and tubo, i.e. cutting the pseudostem at the corm junction and cutting it at 25-30 cm height, respectively, and three extents of parting the corm (whole, half, and quarter) using corms with a diameter of 45 ± 2.9 cm. The experiments revealed that wild enset can be successfully propagated vegetatively in the same way as the cultivated enset. It also revealed that the regeneration process involved callus formation and adventurous bud proliferation from corms only after the apical meristem was removed. Corms of different sizes varied in their capacity for regeneration significantly with a linear increase in regeneration frequency with corm size. With a one cm increase in corm diameter, regeneration frequency increased by 3.138 %. The two populations of wild enset showed non-significant differences in regeneration capacity; however, the achieved regeneration was generally analogous to that observed among cultivated enset clones: whole corms resulted in a longer time to emergence and fewer sucker per corm than split corms. Specifically, halved corms emerged significantly (p < 0.05) earlier (71 ± 9 and 75 ± 7 days, for Shebena and Getiba populations, respectively) than whole corms (120 days). Regeneration frequency was higher (75-100%) for split than for whole corms (33-56%). The highest rate of suckering (94 ± 14 per corm) was achieved from quarter corms prepared by cutting the pseudo-stem at the junction. In conclusion, the adventitious bud propagation technique developed by farmers to propagate the cultivated enset can successfully be used for the clonal regeneration of wild enset. We recommend the adoption of this shoot induction to conserve and maintain the rapidly eroding wild enset genetic resources in Ethiopia. In addition, wild enset plants with promising characteristics may be fixed using the method to enrich the gene pool of the cultivated enset.

Plant breeders and conservationist depend on knowledge about the genetic variation of their species of interest. Pisum fulvum, a wild relative of domesticated pea, has attracted attention as a genetic resource for crop improvement, yet little information about its diversity in the wild has been published hitherto. We sampled 15 populations of P. fulvum from Israeli natural habitats and conducted genotyping by sequencing to analyse their genetic diversity and adaptive state. We also attempted to evaluate the species past demography and the prospects of its future reaction to environmental changes. The results suggest that genetic diversity of P. fulvum is low to medium and is distributed between well diverged populations. Surprisingly, with 56 % in the total population the selfing rate was found to be significantly lower than expected from a species that is commonly assumed to be a predominant selfer. We found a strong genetic bottleneck during the last glacial period and only limited patterns of isolation by distance and environment, which explained 13 %-18 % of the genetic variation. Despite the weak signatures of genome-wide IBE, 1,354 markers were significantly correlated with environmental factors, 1,233 of which were located within known genes with a nonsynonymous to synonymous ratio of 0.382. Species distribution modelling depicted an ongoing fragmentation and decreased habitable area over the next 80 years under two different socio-economic pathways. Our results suggest that complex interactions of substantial drift and selection shaped the genome of P. fulvum. Climate changeis likely to cause further erosion of genetic diversity in P. fulvum. Systematic ex-situ conservation may be advisable to safeguard genetic variability for future utilization of this species.