Hybrids between cultivated and wild carrots in natural populations in Denmark

Many cultivated plant species are able to hybridize with related wild plants. However, it is not clear whether their hybrids are able to survive and reproduce outside managed fields, and if cultivar genes introgress into wild populations. In areas where wild carrots co-occur with carrot root-crops, pollen and seeds may flow from two different sources in the fields to the surrounding wild populations: from pure cultivar plants that occasionally flower, and from flowering 'bolters' that originate from hybridizations between wild (male) and cultivated carrots (female) in seed production fields in warmer regions of the world. To test whether hybrids are formed and survive in wild Danish populations, and whether prolonged hybridization has led to introgression of cultivar genes, we collected leaf material from adult individuals growing close to carrot fields and analysed their genotypic composition by AFLP. Four hybrids were identified among the 71 plants analysed, and these were most likely F(2) or backcross individuals, sired by pollen from hybrid bolters. Wild populations close to fields were genetically somewhat more similar to cultivars than wild populations far from fields, suggesting that neutral or beneficial cultivar alleles can introgress into the wild gene pool. Despite generations of improvement and adaptation of cultivar carrots to highly managed field conditions, hybrids can thus sometimes survive in wild populations close to carrot fields, and their genes transfer to wild populations by introgression.     

Introgression and persistence of cultivar alleles in wild carrot (Daucus carota) populations in the United States

Premise

Cultivated species and their wild relatives often hybridize in the wild, and the hybrids can survive and reproduce in some environments. However, it is unclear whether cultivar alleles are permanently incorporated into the wild genomes or whether they are purged by natural selection. This question is key to accurately assessing the risk of escape and spread of cultivar genes into wild populations.

Methods

We used genomic data and population genomic methods to study hybridization and introgression between cultivated and wild carrot (Daucus carota) in the United States. We used single nucleotide polymorphisms (SNPs) obtained via genotyping by sequencing for 450 wild individuals from 29 wild georeferenced populations in seven states and 144 cultivars from the United States, Europe, and Asia.

Results

Cultivated and wild carrot formed two genetically differentiated groups, and evidence of crop–wild admixture was detected in several but not all wild carrot populations in the United States. Two regions were identified where cultivar alleles were present in wild carrots: California and Nantucket Island (Massachusetts). Surprisingly, there was no evidence of introgression in some populations with a long-known history of sympatry with the crop, suggesting that post-hybridization barriers might prevent introgression in some areas.

Conclusions

Our results provide support for the introgression and long-term persistence of cultivar alleles in wild carrots populations. We thus anticipate that the release of genetically engineered (GE) cultivars would lead to the introduction and spread of GE alleles in wild carrot populations.     

An eco-metabolomic study of host plant resistance to Western flower thrips in cultivated,biofortified and wild carrots

Domestication of plants and selection for agronomic traits may reduce plant secondary defence metabolites relative to their ancestors. Carrot (Daucus carota L.) is an economically important vegetable. Recently, carrot was developed as a functional food with additional health-promoting functions. Biofortified carrots contain increased concentrations of chlorogenic acid as an antioxidant. Chlorogenic acid is involved in host plant resistance to Western Flower Thrips (Frankliniella occidentalis), one of the key agri- and horticultural pests worldwide. The objective of this study was to investigate quantitative host plant resistance to thrips in carrot and to identify candidate compounds for constitutive resistance. As such we explored whether cultivated carrot is more vulnerable to herbivore attack compared to wild carrot. We subjected a set of 14 biofortified, cultivated and wild carrot genotypes to thrips infestation. We compared morphological traits and leaf metabolic profiles of the three most resistant and susceptible carrots using nuclear magnetic resonance spectroscopy (NMR). In contrast to our expectation, wild carrots were not more resistant to thrips than cultivated ones. The most thrips resistant carrot was the cultivar Ingot which is known to be tolerant against carrot root fly (Psila rosae). Biofortified carrots were not resistant to thrips. Plant size, leaf area and number of leaf hairs did not differ between resistant and susceptible carrots. The metabolic profiles of the leaves of resistant carrots were significantly different from those of susceptible carrots. The leaves of resistant carrots contained higher amounts of the flavanoid luteolin, the phenylpropanoid sinapic acid and the amino acid β-alanine. The negative effect of these compounds on thrips was confirmed using in-vitro bioassays. Our results have potential implications for carrot breeders. The natural variation of metabolites present in cultivated carrots can be used for improvement of thrips resistance. This is especially promising in view of the candidate compounds we identified since they do not only confer a negative effect on thrips but as antioxidants also play an important role in the improvement of human health.     

Historical and contemporary gene dispersal in wild carrot (Daucus carota ssp. carota) populations

Background and Aims

Wild carrot is the ancestor of cultivated carrot and is the most important gene pool for carrot breeding. Transgenic carrot may be released into the environment in the future. The aim of the present study was to determine how far a gene can disperse in wild carrot populations, facilitating risk assessment and management of transgene introgression from cultivated to wild carrots and helping to design sampling strategies for germplasm collections.

Methods

Wild carrots were sampled from Meijendel and Alkmaar in The Netherlands and genotyped with 12 microsatellite markers. Spatial autocorrelation analyses were used to detect spatial genetic structures (SGSs). Historical gene dispersal estimates were based on an isolation by distance model. Mating system and contemporary pollen dispersal were estimated using 437 offspring of 20 mothers with different spatial distances and a correlated paternity analysis in the Meijendel population.

Key Results

Significant SGSs are found in both populations and they are not significantly different from each other. Combined SGS analysis indicated significant positive genetic correlations up to 27 m. Historical gene dispersal σ_g and neighbourhood size N_b were estimated to be 4–12 m [95 % confidence interval (CI): 3–25] and 42–73 plants (95 % CI: 28–322) in Meijendel and 10–31 m (95 % CI: 7–∞) and 57–198 plants (95 % CI: 28–∞) in Alkmaar with longer gene dispersal in lower density populations. Contemporary pollen dispersal follows a fat-tailed exponential-power distribution, implying pollen of wild carrots could be dispersed by insects over long distance. The estimated outcrossing rate was 96 %.

Conclusions

SGSs in wild carrots may be the result of high outcrossing, restricted seed dispersal and long-distance pollen dispersal. High outcrossing and long-distance pollen dispersal suggest high frequency of transgene flow might occur from cultivated to wild carrots and that they could easily spread within and between populations.     

Morphological markers for the detection of introgression from cultivated into wild carrot (Daucus carota L.) reveal dominant domestication traits

Hybridisation and subsequent introgression have recently received much attention in the context of genetically modified crops. But crop–wild hybrid detection in the field can be difficult, as most domestication traits seem to be recessive, and the hybrid phenotype may also depend on the direction of the cross or environmental factors. Our aim was to develop a reliable set of morphological markers that differ between two wild and 13 cultivated carrots (Daucus carota L.) and to evaluate their inheritance in hybrid lines. We then examined these morphological markers in four F1 hybrids obtained by fertilising plants from the two wild accessions with pollen from two common carrot cultivars. Of the 16 traits that differed between the two carrot subspecies, three took intermediate values in the hybrids, eight resembled the cultivar parent (dominant domestication traits), two resembled the wild parent (domestication traits recessive), and three were not significant or growth condition‐dependent. Root:shoot ratio was seven times higher for cultivars than for wild plants, while still attaining equivalent total dry weight, which shows that dry matter production by the shoot is much higher in cultivars than in wild plants. High root:shoot ratios were also present in the hybrids. While we found no maternal effects, the type of cultivar used for pollination had an impact on hybrid characteristics. The morphological markers developed here provide insights into the mode of inheritance of ecologically relevant traits and can be useful for pre‐screening wild populations for hybrid detection prior to genetic analysis.     

Dissimilarity of contemporary and historical gene flow in a wild carrot (Daucus carota) metapopulation under contrasting levels of human disturbance: implications for risk assessment and management of transgene introgression

Background and Aims

Transgene introgression from crops into wild relatives may increase the resistance of wild plants to herbicides, insects, etc. The chance of transgene introgression depends not only on the rate of hybridization and the establishment of hybrids in local wild populations, but also on the metapopulation dynamics of the wild relative. The aim of the study was to estimate gene flow in a metapopulation for assessing and managing the risks of transgene introgression.

Methods

Wild carrots (Daucus carota) were sampled from 12 patches in a metapopulation. Eleven microsatellites were used to genotype wild carrots. Genetic structure was estimated based on the F_ST statistic. Contemporary (over the last several generations) and historical (over many generations) gene flow was estimated with assignment and coalescent methods, respectively.

Key Results

The genetic structure in the wild carrot metapopulation was moderate (F_ST = 0·082) and most of the genetic variation resided within patches. A pattern of isolation by distance was detected, suggesting that most of the gene flow occurred between neighbouring patches (≤1 km). The mean contemporary gene flow was 5 times higher than the historical estimate, and the correlation between them was very low. Moreover, the contemporary gene flow in roadsides was twice that in a nature reserve, and the correlation between contemporary and historical estimates was much higher in the nature reserve. Mowing of roadsides may contribute to the increase in contemporary gene flow. Simulations demonstrated that the higher contemporary gene flow could accelerate the process of transgene introgression in the metapopulation.

Conclusions

Human disturbance such as mowing may alter gene flow patterns in wild populations, affecting the metapopulation dynamics of wild plants and the processes of transgene introgression in the metapopulation. The risk assessment and management of transgene introgression and the control of weeds need to take metapopulation dynamics into consideration.     

Further studies relating chlorogenic acid concentration in carrots to carrot fly damage

The effects of first generation carrot fly larval damage on chlorogenic acid concentration in carrots was investigated in a field experiment at Wellesbourne in 1985. In a separate experiment carrots grown in the absence of a resident population of carrot fly were also analysed for chlorogenic acid; these carrots maintained low concentrations of chlorogenic acid through the summer and autumn until low ground temperatures occurred from November to January. The relationship between chlorogenic acid concentration and damage by the first generation of carrot fly was described by a similar model to the one derived previously for late-generation damage but without the cultivar dependence. This may have been because first generation damage takes place in mid-summer when soil temperature is not sufficiently low for differential chlorogenic acid production by carrot cultivars. The model supports the hypothesis that carrot fly damage increases chlorogenic acid production which subsequently encourages further attack. The increase in acid production due to the low winter temperature may be the mechanism which, in turn, induced a differential cultivar response in carrots harvested during the winter.     

Use of the advanced backcross-QTL method to transfer seed mineral accumulation nutrition traits from wild to Andean cultivated common beans

Iron deficiency anemia and zinc deficiency are major health concerns across the world and can be addressed by biofortification breeding of higher mineral concentration in staple crops, such as common bean. Wild common beans have for the most part had higher average seed mineral concentration than cultivars of this species but have small un-commercial seeds. A logical approach for the transfer of the seed mineral trait from wild beans to cultivated beans is through the advanced backcross breeding approach. The goal of this study was to analyze a population of 138 BC(2)F(3:5) introgression lines derived from the very high iron wild genotype G10022 backcrossed into the genetic background of the commercial-type variety 'Cerinza', a large-red seeded bush bean cultivar of the Andean genepool. In addition to measuring seed mineral accumulation traits and the quantitative trait loci (QTL) controlling these traits we were interested in simultaneously testing the adaptation of the introgression lines in two replicated yield trials. We found the cross to have high polymorphism and constructed an anchored microsatellite map for the population that was 1,554-cM long and covered all 11 linkage groups of the common bean genome. Through composite interval mapping (CIM) and single point analysis (SPA), we identified associations of markers and mineral traits on b01, b06, b07, b08, b10 and b11 for seed iron concentration, and markers on b01, b04 and b10 for seed zinc concentration. The b07 and b08 QTL aligned with previous QTL for iron concentration. A large number of QTL were found for seed weight (9 with CIM and 36 with SPA analysis) and correlations between seed size and mineral content affected the identification of iron and zinc contents' QTL on many linkage groups. Segregation distortion around domestication genes made some areas difficult to introgress. However, in conclusion, the advanced backcross program produced some introgression lines with high mineral accumulation traits using a wild donor parent.     

Exploitation of the resistance to carrot fly in the wild carrot species Daucus capillifolius

Under field conditions a wild Daucus species from Libya, D. capillifolius, supported less than one tenth as many carrot flies (Psila rosae) as the susceptible carrot cultivar Danvers Half Long 126. Breeding lines developed from crosses between D. capillifolius and three different carrot types were grown in a series of field experiments at Wellesbourne between 1980 and 1989. Each year selections were made for agronomic quality and/or for increased resistance to carrot fly. The programme produced lines which for size, shape and colour represented most of the commercially-important carrot types. Some of these lines were also significantly more resistant to carrot fly than selections from the partially-resistant cv. Sytan. However, the best lines were not as resistant as the wild parent. The highest quality resistant lines were sold to seed companies for variety production.     

Effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism in seedlings of wheat cultivars

The effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism (alpha and beta amylases, sucrose phosphate synthase, sucrose synthase, acid and alkaline invertases) in wheat (Triticum aestivum L.) was investigated in the seedlings of drought-sensitive (PBW 343) and drought-tolerant (C 306) cultivars. The water deficit was induced by adding 6% mannitol (water potential -0.815 Mpa) in the growth medium. The water deficit reduced starch content in the shoots of tolerant seedlings as compared to the sensitive ones, but increased sucrose content in the shoots and roots of tolerant seedlings, indicating their protective role during stress conditions. It also decreased the alpha-amylase activity in the endosperm of seedlings of both the cultivars, but increased alpha and beta amylase activities in the shoots of tolerant ones. Sucrose phosphate synthase (SPS) activity showed a significant increase at 6 days of seedling growth (DSG) in the shoots of stressed seedlings of tolerant cultivar. However, SPS activity in the roots of stressed seedlings of sensitive cultivar was very low at 4 DSG and appeared significantly only at day 6. Sucrose synthase (SS) activity was lower in the shoots and roots of stressed seedlings of tolerant cultivar than sensitive ones at early stage of seedling growth. Higher acid invertase activity in the shoots of seedlings of tolerant cultivar appeared to be a unique characteristic of this cultivar for stress tolerance. Alkaline invertase activity, although affected under water deficit conditions, but was too low as compared to acid invertase activity to cause any significant affect on sucrose hydrolysis. In conclusion, higher sucrose content with high SPS and low acid invertase and SS activities in the roots under water deficit conditions could be responsible for drought tolerance of C 306.     

The role of domestication and maternal effects on seed traits of crop–wild sunflower hybrids (Helianthus annuus)

Hybridisation between crops and their wild relatives may promote the evolution of weeds. Seed germination and dormancy are the earliest life‐history traits and are highly influenced by the maternal parent. However, the ecological role of the maternal effect on seed traits in the evolution of crop–wild hybrids has received little attention. In this study, we test the relative importance of maternal and hybridisation effects on seed traits of the first generation of crop–wild sunflower hybrids (Helianthus annuus). Seed germination was tested in two wild populations with contrasting dormancy, two cultivated materials and their reciprocal crosses at four different times after harvest and three different temperatures. Seed germination at each of the four times, after ripening response and secondary dormancy were recorded along with four morphological traits. Additionally, the pericarp anatomy was analysed with light and scanning electron microscopy. We observed strong maternal effects on all seed traits. Seed germination, morphology and pericarp anatomy differed largely between the crop and wild seeds and these traits in the crop–wild hybrids resembled their female parent. Slight but significant hybridisation effects were observed in germination, mainly in seeds produced on wild plants. Crop hybridisation changed seed germination, the after ripening response and secondary dormancy in the crop direction. Morphological and anatomical traits associated with domestication strongly correlated with the observed differences in seed germination and dormancy in crop–wild sunflower hybrids. The large maternal effects along with the evolutionary divergence in seed traits were responsible for the large phenotypic differences observed in crop–wild hybrids with the same genetic composition. Wild‐like seed traits of hybrids suggest that there are no barriers to crop gene introgression at the seed level whereas crop‐like seed traits could be strongly selected against, conditioning the selection of traits expressed later in the life cycle and in the next generations.     

Fine-scale phylogenetic structure and major events in the history of the current wild soybean (Glycine soja) and taxonomic assignment of semi-wild type (Glycine gracilis Skvortz.) within the Chinese subgenus Soja

Wild and cultivated species of soybeans have coexisted for 5000 years in China. Despite this long history, there is very little information on the genetic relationship of Glycine soja and G. max. To gain insight into the major events in the history of the subgenus Soja, we examined 20 simple sequence repeat (SSR) markers of a large number of accessions (910). The results showed no significant differences between wild and semi-wild soybeans in genetic diversity but significant differences between G. soja and G. max. Ancestry and cluster analyses revealed that semi-wild soybeans should belong to the wild category and not to G. max. Our results also showed that differentiation had occurred not only among G. soja, G. gracilis, and G. max but also within G. soja and within G. gracilis. Glycine soja had 3 clear genetic categories: typical small-seeded (≤2.0 g 100-seed weight), dual-origin middle-seeded (2.0-2.5 g), and large-seeded plants (2.51-3.0 g). These last were genetically close to G. gracilis, their defining some traits having been acquired mainly by introgression from soybeans. Small-seeded G. gracilis (3.01-3.5 g) were genetically different from larger seeded ones (from 3.51 to 4.0 to over 10 g). Seed size predominated over seed coat color in evolutionary degree. Typical and large-seeded G. soja were found to have 0.7% and 12% introgressive cultivar genes, respectively. The genetic boundary of G. gracilis was at the range of 2.51-3.0 g of G. soja. In the great majority of wild accessions, traits such as white flowers, gray pubescences, no-seed bloom, and colored seed coats were likely introgressive from domesticated soybeans.     

Electrophoretic variation in glutamate dehydrogenase and other isozymes in wild carrot cells cultured in the presence and absence of 2,4-dichlorophenoxyacetic acid

Summary Electrophoretic analysis of isozymal differences was performed with extracts of wild carrot (Daucus carota L.) cells, grown in the presence and absence of 2,4-dichlorophenoxyacetic acid (2,4-D). There were no differences in the patterns of malate dehydrogenase, acid phosphatase, aspartate aminotransferase, and γ-glutamyl transferase. Quantitative differences in peroxidase isozymes were detected, the plus 2,4-D cultures having lower activities. Esterase patterns were similar, but there were differences in individual isozyme activities and an additional form present in the minus 2,4-D cells. the greatest differences were in patterns of glutamate dehydrogenase with the minus 2,4-D cultures containing only the slowly migrating isozymes. The changes in glutamate dehydrogenase, as revealed by isozyme changes, together with the requirement for ammonia in embryogenesis, suggests that this enzyme may be associated with differentiation in wild carrot cells.     

新疆核桃4个栽培品种及野生种的遗传多样性分析

以新疆地区种植的4个核桃(Juglans regia Linn.)栽培品种(包括新栽培品种'温185'和'新新2'以及老栽培品种'新丰'和'扎343')及巩留野核桃自然保护区生长的野生核桃为研究对象,对其cpDNA的psbK-psbI区间和mtDNA的COX2 intronⅠ区间以及nrDNA的ITS和ETS区间的DNA片段序列进行了比较分析,并对其MP、ML和UPGMA系统发育树进行了分析;此外,还基于SSR分子标记结果对其进行了遗传多样性指数、UPGMA系统发育树和遗传分组分析.结果表明:野生种与4个栽培品种的cpDNA和mtDNA片段序列无碱基变异,而其nrDNA的片段序列却存在3个碱基变异,但4个栽培品种间无碱基变异.以麻核桃(J.hopeiensis Hu)为外类群,基于上述4个DNA片段序列构建的MP、ML和UPGMA系统发育树的聚类结果一致,均表现为4个栽培品种聚为一组,而野生种和麻核桃则分别单独聚为一组.野生种的观测杂合度、预期杂合度和固定指数分别为0.383、0.448和0.153,4个栽培品种的上述3个遗传多样性指数分别为0.428~0.576、0.423~0.619和-0.043~0.234.基于SSR分子标记结果的UPGMA系统发育树和分组数为5的遗传分组结果均表明:野生种和品种'温185'分别单独为一组;品种'新新2'和'新丰'为一组;而品种'扎343'也单独为一组,但与品种'新新2'和'新丰'遗传关系较近.遗传分组结果还表明:分组数为3更利于明确品种'扎343'的分组地位,此时,其与品种'新新2'和'新丰'为一组.综合分析结果表明:核桃4个栽培品种间的遗传差异较小,且老栽培品种的遗传多样性总体上高于新栽培品种;野生种与栽培品种间具有明显的遗传差异,说明在育种或栽培过程中核桃种质资源的遗传多样性可能会逐渐降低,并且,该野生种可为核桃的分子育种提供天然的基因库资源.     

Diversity and analysis of sequences encoded by arcelin genes from Indian wild pulses resistant to bruchids

Wild pulse accessions are considered a vital source of genes for insect resistance for crop improvement programmes. Wild pulses resistant to infestation towards the bruchid insect pest, Callosobruchus maculatus from South India were chosen to screen the existence of potent insecticidal protein, arcelin from APA locus (Arcelin/Phytohemagglutinin/α-Amylase inhibitor) to ascertain their nature and functional diversity without any specific indication for insect resistant factors. The DNA sequence coding for arcelin from various species of wild pulses were amplified, sequenced and deduced to their protein sequences. These protein sequences were examined physico-chemically using several bioinformatics tools and docked with various sugars to resolve the nature of arcelin molecules. Results indicated the presence of significant differences in the properties of arcelin molecules from various species of Indian wild pulses with their amino acid sequences, several physico-chemical properties and binding ability with sugars. The differences observed on these arcelin molecules from diverse wild pulses are predicted to provide a prospective insect pest control factors.     

The stability of accumulated anthocyanin in suspension cultures of the parental line and high and low accumulating subclones of wild carrot

Abhibition of the accumulation of anthocyanin but not of growth in wild carrot cultures was achieved using 2-amino-oxyphenylpropionic acid or phenylpropiolic acid. The concentration of inhibitor required to achieve this differential effect increased with increased culture density.By several additions of inhibitor at differential times, anthocyanin accumulation in the parental culture and in high-and low-acculmulating subclones could be completely inhibited. Under these conditions the level of anthocyanin in the cultures remained unchanged. These data show that the accumulated anthocyanin is not metabolized and that differences in the rate of degradation of anthocyanin does not account for the differences in anthocyanin accumulation by the subclones examined.     

The consistency of the resistance of eight carrot cultivars to carrot fly attack at several centres in Europe

Eight carrot cultivars representing the range of resistance to carrot fly damage as assessed in England were grown at 12 sites in five European countries in 1977 and in 1978. The trials provided evidence of agreement with the findings in England for both years in the ranking of cultivars with respect to their resistance to carrot fly damage. The Nantes cultivar Clause's Sytan Original was the most resistant cultivar tested. Consistent results were obtained from sites where carrots were assessed at harvest and carrot fly attack was severe. A significant interaction between sites and cultivars was largely due to sites where carrots were stored prior to assessment.     

Frost sensitivity of hybrids between wild and cultivated carrots

Frost could potentially limit the survival ofhybrids between cultivated and wild carrots innatural habitats, and thereby the likelihoodthat (trans)genes spread from the crop to wildplants. To test this, cultivated, wild carrotsand their hybrids were exposed to differentfrost treatments. Hybrids survivedsignificantly less than the wild carrots, butonly slightly better than the cultivars,indicating that frost will indeed limit theirsurvival.     

Glucosinolates of wild and cultivated Brassica species

Major differences were found in the occurrence of glucosinolates in the leaves of different wild species of the Brassica oleracea (2n = 18) group. The distribution of glucosinolates supports the present classification with the exception of the inclusion of both Tunisian and Sardinian populations within B. insuloris. The results also support the hypothesis of a multiphyletic origin of cultivated forms of B. oleracea from a number of wild species. In contrast, wild populations of B. rapa had very similar glucosinolate contents. Wild populations had a considerably higher total glucosinolate content than cultivars which may limit their usefulness in breeding programmes.