**3. Plant domestication in the 21st century: A case study with PEI wild rosehips**

One of the most recent and successful domestication of a wild species is that of the North American ginseng [57]. Similar to ginseng, interests in wild rosehip products are increasing worldwide due to its nutraceutical and natural health products properties [13]. With aging and changing eating lifestyles, the incidence of chronic diseases is increasing worldwide. Despite success achieved in fighting these diseases, prevention measures have become top priorities for citizens and public health systems. Recently, increasing interest has been expressed in plant natural products as preventative agents. Hence, plant product preparations such as those from rosehip have been used as food and medicine for centuries. The genus *Rosa* contains more than 150 species. They are widespread in North America within the *Cinnamo‐ mae* section and are renowned for the vitamin C content [58-61]. Although formulations from *Rosa canina* have been associated with the treatment and symptom reduction of inflammation and arthritis, the vast majority of wild rose species are fully unexplored for their heath potential. To date, most of the reported studies were focused mainly on *Rosa* species within the *Caninae* section which comprises 20 – 30 *Rosa* species known as dogroses [18, 42] and is currently the focus of major domestication research programs for the production and com‐ mercialisation of rosehips (fruits) around the world, particularly in Northern Europe, Germa‐ ny, Turkey, Eastern Europe and Chile [13]. So far, less emphasis has been made on *Rosa* species belonging to *R. carolina* complex within the *Cinnamomae* section and the rosehips production from the eastern North American native wild roses is new and emerging [55, 56]. This section deals with the genetic diversity of PEI wild rosehips, the challenges associated with their domestication as well as the agronomic practices that could ensure an economic production.

#### **3.1. Introduction to the genus** *Rosa*

the societal needs. In the present global economy, the scale of demands for any good has increased and the trade has become multidirectional (selling in all part of globe) with multiple layers (one product could be found in many other products as additive or supplement) (Table 1). Thus, probing the genetic diversity of a plant species which end-product would satisfy these new needs both in terms of quality, quantity, sustainability and stability has become the new challenge for plant products developers. Hence, the need for well characterized germplasm with stable and preserved genetic identity is becoming the landmark for todays and tomor‐ row's natural product designers and developers. Therefore, sophisticated molecular tools [51, 52] as well as mass tissue culture and plant propagation tools are being employed to insure

**Ancient domestication Domestication in the 21st century References**

sustainability

Food, clothing, energy, health, life quality,

flavour, energy, metabolite profiles,

and fields, large commercial fields, high throughput management, human and animal

force and mechanization

Ecosystem Complex Simple [53]

Value chain Self, local consumption, Global, processing, distribution and marketing

Yield Low High [28, 55, 56]

networks

**Table 1.** Comparative pathways of ancient and modern plant domestication processes: purposes, tools, and

**3. Plant domestication in the 21st century: A case study with PEI wild**

One of the most recent and successful domestication of a wild species is that of the North American ginseng [57]. Similar to ginseng, interests in wild rosehip products are increasing worldwide due to its nutraceutical and natural health products properties [13]. With aging

Morphology, genetic DNA markers, QTLs, taste,

Experimental tubes, growth chambers, greenhouse

[28, 53, 54]

[20, 41, 51, 52]

[28, 53]

stability and sustainability.

Screening methods

expectations

**rosehips**

Purposes Food, medicine clothing,

energy

Production paths Gathering, yards and small

energy, sustainability

Morphology, taste, flavour,

42 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

farms sowing and harvesting, human and animal force

Purity Composite Composite, variety

The genus *Rosa* (*Rosaceae*) originated in the temperate regions of the northern hemisphere, including North America, Europe, Asia, and the Middle East, with the greatest diversity of species found in western China, where it is endemic, and is now widespread all over the globe [18]. With this wide distribution range and the high number of species (more than 150 shrub species), the delimitation of the species bounbaries remained a challenge for taxonomists and molecular biologists [16, 21, 41, 44].

#### **3.2.** *Rosa* **species phylogeny and biodiversity**

#### *3.2.1. Global Rosa species biodiversity and phylogeny*

The taxonomy and breeding system of the genus *Rosa* has been recently reviewed by several authors [13, 16, 21, 38, 49, 62, 63] and the reader is invited to find more details in these treatments. Of particular interests are works reported by Werlemark and Nybom [13] and Macphail and Kevan [21] on one hands, and those by Bruneau et al. [16] and Joly and Bruneau [44] on the other hands, focusing on the European Dogroses from section *Caninae* and the North American *Rosa* species from section *Cinnamomae*, respectively. Wild rose species from these two sections are currently extensively investigated for domestication purposes and commer‐ cial rosehip production [13, 55, 64-67]. As our interest lies mainly in the domestication of North American wild roses, the next section of this review will put more emphasis on the biodiversity and phylogeny of wild rose species commonly encountered in this part of the globe and more specifically in Canada, a country as large as the whole Europe (West and East taken together, excluding the former USSR).

#### *3.2.2. North American Rosa species biodiversity and phylogeny*

Biodiversity of the North American wild roses has been investigated by botanists in the early 1900's. Watson [68], Crepin [69, 70], Erlanson MacFarlane [71, 72] have described and defined 13 - 22 *Rosa* species in North America. This important polymorphism in *Rosa* species, especially in eastern North America, together with hybridization and polyploidy have long been considered as the major causes of taxonomic confusion in the genus [17]. Alfred Rehder (1869-1949) estab‐ lished the first foundation of *Rosa* species taxonomic relationship in a book entitled "*TheManual of Cultivated Trees and Shrubs Hardy in North America Exclusive of the Subtropical and Warmer Tem‐ perate Regions"* published in 1940 [49]. Rehder provided concise physical description, time of flowering, region of native habitat, hardiness zone, distinguishing features and pertinent infor‐ mation on North American roses, and subdivided the genus *Rosa* into 4 subgenera and 10 sec‐ tions, including the *Rosa carolina* L. complex of section *Cinnamomae*. East of the Rocky Mountain, the *Rosa Carolina* complex is composed of five diploid species (*R. blanda*, Ait., *R. foliolosa* Nutt., *R. nitida* Wild., *R. palustris* March., and *R. Woodsii* Lindl.), three tetraploid species (*R. carolina* L., *R. virginiana* Mill., and *R. arkansana* Porter) and one hexaploid/octaploid species (*R. acicularis* Lindl.) which is morphologically distinct from all other species [17]. The taxonomic problems are well known at the diploid level, where some species hybridize and are also morphologically difficult to distinguish (which is particular true for *R. blanda* and *R. woodsii*), but are even more acute at the polyploidy level. *Rosa carolina* which is widespread East of the Mississipi river hy‐ bridizes with *R. Arkansana* in the western part of its distribution [71] but also in the East with *R. virginiana*. Moreover, the morphological similarity cuts across ploidy levels and no single mor‐ phological character can be used to distinguish one species to another [17]. Thanks to molecular tools (AFLP, SNP), haplotype network analysis using statistical parsimony, genealogical ap‐ proach, and multivariate analysis of 25 morphological characters including ploidy determina‐ tion based on stomatal guard cell lengths, Joly et al. [17] and Joly and Bruneau [44] determined four species at the diploid level and that were separated into 2 groups in the east of the Rocky Mountains: one group consists of *R. blanda - R. woodsii* (which were indistinguishable and should be considered as a single species), and the other group is consisted of *R. foliolosa*, *R. nitida*, and *R. palustris*. The authors also determined 3 species at the polyploid level: *R. arkansana*, *R. car‐ olina*, *R. virginiana*, with evidence of hybridization between them. The diploids that are involved in the origins of the polyploid species in that region were also proposed. For Joly et al. [17], only diploids east of the Rocky Mountains are involved in the origins of polyploids. *Rosa arkansana* is derived from the *blanda-woodsii* group, *R. virginiana* originated from the *foliolosa- nitida-palustris* group, and *R. carolina* is derived from a hybrid between the two diploid groups. Thus, for wild rose species domestication and commercial production purposes in the Canadian Maritimes where both North American native wild species of the *R. carolina* complex grow in sympatry and also along with naturalized species such as *R. rugosa* or other members of dogroses (Figure 1), a careful species determination as well as genotypic identification of collected germplasm for propagation are of critical importance to ensure, genetic purity and traceability.

Genetic, Agronomy, and Metabolomics of Prince Edwards Island Wild Rose Collection and Promise for Cultivar Development http://dx.doi.org/10.5772/54688 45

**Figure 1.** Diversity of rosehip morphology in the Atlantic Canada landscape. A, typical morphology PEI grown rosehip. B, morphological feature of a naturalized rosehip to Atlantic Canada.

#### *3.2.3. Genetic and Metabolite diversity within the Prince Edward Island's field collection*

Using SSR markers [20] and single nucleotide polymorphisms analysis, our group has assessed the genetic diversity within 30 ecotypes under cultivation and identified three major clusters, with cluster 2 and 3 showing 2 and 3 sub-clusters, respectively [65, 73]. The metabolite profiles in the flesh, seed, and fuzz for anthocyanins, flavonols, tilirosides which is a potent antidiabetic compound, tannins and fatty acids were also determined from the 30 ecotypes [65, 73]. The level of anthocyanin was very low in all ecotypes, with only one ecotype showing a level that was 30-40 % higher compared to the average. A large diversity was observed for flavonols and tiliroside among ecotypes. Only 4 ecotypes had a high content for both flavonols and tiliroside in the analyzed tissues (Ghose et al, submitted). One ecotype showed 18:3 level as high as 41.2%. The data suggests that it is possible to select and propagate a given ecotype for its unique metabolite profile for commercial and drug production [65, 73].

#### **3.3. Domestication and end uses**

Roses have been domesticated by man first for the beauty of their flower and incorporated in many cultural and political practices [74] and are now encountered on all continents, climates, and market places. Nonetheless, the medicinal uses of rose leaves, flowers and fruits were also widespread in human history [13, 54, 75-78].

#### *3.3.1. Flower roses*

*3.2.2. North American Rosa species biodiversity and phylogeny*

44 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

Biodiversity of the North American wild roses has been investigated by botanists in the early 1900's. Watson [68], Crepin [69, 70], Erlanson MacFarlane [71, 72] have described and defined 13 - 22 *Rosa* species in North America. This important polymorphism in *Rosa* species, especially in eastern North America, together with hybridization and polyploidy have long been considered as the major causes of taxonomic confusion in the genus [17]. Alfred Rehder (1869-1949) estab‐ lished the first foundation of *Rosa* species taxonomic relationship in a book entitled "*TheManual of Cultivated Trees and Shrubs Hardy in North America Exclusive of the Subtropical and Warmer Tem‐ perate Regions"* published in 1940 [49]. Rehder provided concise physical description, time of flowering, region of native habitat, hardiness zone, distinguishing features and pertinent infor‐ mation on North American roses, and subdivided the genus *Rosa* into 4 subgenera and 10 sec‐ tions, including the *Rosa carolina* L. complex of section *Cinnamomae*. East of the Rocky Mountain, the *Rosa Carolina* complex is composed of five diploid species (*R. blanda*, Ait., *R. foliolosa* Nutt., *R. nitida* Wild., *R. palustris* March., and *R. Woodsii* Lindl.), three tetraploid species (*R. carolina* L., *R. virginiana* Mill., and *R. arkansana* Porter) and one hexaploid/octaploid species (*R. acicularis* Lindl.) which is morphologically distinct from all other species [17]. The taxonomic problems are well known at the diploid level, where some species hybridize and are also morphologically difficult to distinguish (which is particular true for *R. blanda* and *R. woodsii*), but are even more acute at the polyploidy level. *Rosa carolina* which is widespread East of the Mississipi river hy‐ bridizes with *R. Arkansana* in the western part of its distribution [71] but also in the East with *R. virginiana*. Moreover, the morphological similarity cuts across ploidy levels and no single mor‐ phological character can be used to distinguish one species to another [17]. Thanks to molecular tools (AFLP, SNP), haplotype network analysis using statistical parsimony, genealogical ap‐ proach, and multivariate analysis of 25 morphological characters including ploidy determina‐ tion based on stomatal guard cell lengths, Joly et al. [17] and Joly and Bruneau [44] determined four species at the diploid level and that were separated into 2 groups in the east of the Rocky Mountains: one group consists of *R. blanda - R. woodsii* (which were indistinguishable and should be considered as a single species), and the other group is consisted of *R. foliolosa*, *R. nitida*, and *R. palustris*. The authors also determined 3 species at the polyploid level: *R. arkansana*, *R. car‐ olina*, *R. virginiana*, with evidence of hybridization between them. The diploids that are involved in the origins of the polyploid species in that region were also proposed. For Joly et al. [17], only diploids east of the Rocky Mountains are involved in the origins of polyploids. *Rosa arkansana* is derived from the *blanda-woodsii* group, *R. virginiana* originated from the *foliolosa- nitida-palustris* group, and *R. carolina* is derived from a hybrid between the two diploid groups. Thus, for wild rose species domestication and commercial production purposes in the Canadian Maritimes where both North American native wild species of the *R. carolina* complex grow in sympatry and also along with naturalized species such as *R. rugosa* or other members of dogroses (Figure 1), a careful species determination as well as genotypic identification of collected germplasm for

propagation are of critical importance to ensure, genetic purity and traceability.

The best known uses for roses are their flowers as ornamental on tables, in home backyards, pub‐ lic gardens and spaces. Historically, only very few wild rose species (at most 5 to 11 species) have been involved as parents in the today flower roses. One example of using native rose species in North America is related to the Parkland Rose series developed at AAFC in Morden, Manitoba. These flower roses are hardy, winter resistant and some of these rose varieties involve in their pedigree *R. Arkansana* which is encountered east of the Rocky Mountain in Canada. Beside, its or‐ namental features, rose flowers are valuable for the cosmetic industry [75, 76, 78].

#### *3.3.2. Wild rosehips*

The fruits of roses, the hips, have been highly regarded as important food and medicinal sources [13, 54, 79]. Rosehip is appreciated as traditional vitamin C rich soup in Sweden where the de‐ mand is particularly high [80]. Its flesh and seeds have been used in concoctions and tonics for various ailments, including the use as laxative and diuretic, against common cold, gastroinstes‐ tinal disorders, gastric ulcers [77, 81, 82], and anti-inflammatory diseases such as arthritis [83]. A review on the major chemical components of dogrose hips from was recently made by Werle‐ mark [13]. However, a marked variation in chemical composition is associated with species, genotypes, and environments in which the plants evolve. For example, Melville and Pyke [84] found a weak correlation between latitude and vitamin C content of British rosehip populations from Scotland and England. Similarly, Werlemark [13] hypothesised that rosehips produced in a colder climate, especially with colder summer, may have higher vitamin C content compared to those that have been maturing in a warmer climate and also anticipated that local variations in precipitations and temperatures during summer may affect the chemical content of rosehips. It is reasonable to assume that, with different species and cooler summer and fall (Table 2), the Canadian Maritime wild rose species would show different chemical composition, especially in terms of relative amount when compared to their European and South American counterparts. By comparing some rosehip samples from Prince Edward Island, Denmark, Chile and South Af‐ rica, our group observed differences between origins, especially with regards to total oil content and fatty acid profiles (Figure 2). Nonetheless, sample preparation (harvesting time and condi‐ tioning) can also be a major source of variation. It will be of interest to compare the chemical composition of rosehips collected in each of these regions during the same summer or fall for ob‐ taining factual and conclusive answers to these assumptions.

**Figure 2.** Comparative study of rosehip samples from Prince Edward Island, Denmark, Chile, and South Africa.

#### Genetic, Agronomy, and Metabolomics of Prince Edwards Island Wild Rose Collection and Promise for Cultivar Development http://dx.doi.org/10.5772/54688 47

Rosehip seed contains pretty well balanced omega-6 (18:2) / omega-3 (18:3) fatty acid ratio and also shows relatively high level of oleic acid as compared to olive and canola oils that are rich in oleic acid but low in both linoleic and linolenic acids (Figure 3). As genetic variability for fatty acid composition has been observed in PEI wild roses (Ghose et al, submitted) and the seed oil content is relatively low, breeding efforts could contribute to increase the oil content.

pedigree *R. Arkansana* which is encountered east of the Rocky Mountain in Canada. Beside, its or‐

The fruits of roses, the hips, have been highly regarded as important food and medicinal sources [13, 54, 79]. Rosehip is appreciated as traditional vitamin C rich soup in Sweden where the de‐ mand is particularly high [80]. Its flesh and seeds have been used in concoctions and tonics for various ailments, including the use as laxative and diuretic, against common cold, gastroinstes‐ tinal disorders, gastric ulcers [77, 81, 82], and anti-inflammatory diseases such as arthritis [83]. A review on the major chemical components of dogrose hips from was recently made by Werle‐ mark [13]. However, a marked variation in chemical composition is associated with species, genotypes, and environments in which the plants evolve. For example, Melville and Pyke [84] found a weak correlation between latitude and vitamin C content of British rosehip populations from Scotland and England. Similarly, Werlemark [13] hypothesised that rosehips produced in a colder climate, especially with colder summer, may have higher vitamin C content compared to those that have been maturing in a warmer climate and also anticipated that local variations in precipitations and temperatures during summer may affect the chemical content of rosehips. It is reasonable to assume that, with different species and cooler summer and fall (Table 2), the Canadian Maritime wild rose species would show different chemical composition, especially in terms of relative amount when compared to their European and South American counterparts. By comparing some rosehip samples from Prince Edward Island, Denmark, Chile and South Af‐ rica, our group observed differences between origins, especially with regards to total oil content and fatty acid profiles (Figure 2). Nonetheless, sample preparation (harvesting time and condi‐ tioning) can also be a major source of variation. It will be of interest to compare the chemical composition of rosehips collected in each of these regions during the same summer or fall for ob‐

namental features, rose flowers are valuable for the cosmetic industry [75, 76, 78].

46 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

taining factual and conclusive answers to these assumptions.

**Figure 2.** Comparative study of rosehip samples from Prince Edward Island, Denmark, Chile, and South Africa.

*3.3.2. Wild rosehips*

**Figure 3.** Comparative fatty acid profile of rosehip with three oilseed crops.

#### *3.3.2.1. Agronomy*

Although a high value was recognized to rosehip throughout centuries, it is only recently that the wild roses are being domesticated and cultivated for their fruits and to develop agronomic practices that ensure an economic production of the hips [28, 51, 52, 56, 77, 85]. However, due to the diversity of species, genotypes, soils and climates, different agronomic practices are being implemented and tested in different regions, including Denmark, Turkey, Bulgaria, Chile and Canada. Whereas Chilean started their trials by developing a nursery built on the "Tunnel" greenhouse model with a capacity to accommodate 15.000 cuttings, under an irrigation system with nebulizers to reduce temperature and humidification before a devel‐ opmental stage in the fields, the Danish, Swedish and Canadian choose to established field trials using wild cutting, spacing, density and nutrient management trials [28, 55]. In Sweden, the germplasm used were mostly concentrated on the Scandinavian *Rosa* species of section *Caninae* especially, *R. dumalis, R. rubiginosa* and their interspecific hybrids [86] whereas Danish rosehips are produced mainly from *R. canina* (www.hyben-vital.com) although it may also involve other Scandinavian species. In Chile, the current production is mainly focused on wild hand-harvested hips from uncharacterized and naturalized species introduced to south America by Spanish and is mostly a mixture of *R. rubiginosa, R. canina, R. moschata* and many other species found in western Europe [66]. In Prince Edwards Island, (Canada), current recent genetic study based on 30 wild ecotypes collected from this province suggested that all accessions currently under field trial are from *R. virginiana* and its natural hybrids with *R. Carolina* (Ghose et al, submitted). At present, very few cultivars have been named and released for commercial fruit production. One cultivar, the cultivar "Mechthilde von Neuerburg" derived from *R. rubiginosa* was reported in Germany. Two cultivars (Sylwia and Sylwana) derived from *R. canina* were reported in Poland, whereas cultivar Plovdiv 1 from *R. canina*, and cultivar Karpatia from *R. villosa* were reported in Bulgaria and Slovakia, respectively [13]. For all of these semi-domesticated wild rosehips, it is not known or reported whether the ongoing domestication process has already impacted on some of the phenotypic traits such fruit size, fruit setting or metabolite profile. By comparing the pomology characteristics of 5 wild rosehip ecotypes growing in the wild or in the field settings, we observed that the field setting contributed to increase the fruits size and delayed the maturity when compared with growing in the wild, suggesting an occurrence of a domestication syndrome for these traits (Fofana, personal observation). However, no significant difference was found between the two environments for the number of seed in each of the ecotype.

#### *3.3.2.1.1. Soils and climates*

Although originally native to temperate regions of the globe, roses have adapted to warmer regions and grow well now in very diversified habitats and soil types [13, 79]. The soil should be well drained though and not heavy. Species preference for soil type has nonetheless been reported. *R. villosa* was reported to grow better in a dry soil with low calcium content whereas *R. canina* and *R. dumalis* prefer more calcareous soil. *R. rubiginosa* also prefers more calcium and grows well in a relatively heavy soil [13]. *R. palustris* grows in marshes and *R. nitida* in bogs. Similarly, *R. virginiana* likes salt marshes and salty soils (Joly, personal communications). In Prince Edwards Island province (Canada), wild rosehips are found in a variety of habitats including hedgerows, wet and dry pastures, thickets, swamps and uplands in dry orthic humoferric Podzol sandy soils [55]. In hard winter climates such as Canada, plant survival rate in the field setting can vary from genotype to genotype and for the same genotype, plastic coverage has been shown to increase the winter survival rate (Figure 4).

**Figure 4.** Effect of planting beds coverage with plastic on winter survival.


**Table 2.** Comparison of average temperature and precipitations during summer and fall in major rosehip production countries.

#### *3.3.2.1.2. Fertilization*

*3.3.2.1. Agronomy*

Although a high value was recognized to rosehip throughout centuries, it is only recently that the wild roses are being domesticated and cultivated for their fruits and to develop agronomic practices that ensure an economic production of the hips [28, 51, 52, 56, 77, 85]. However, due to the diversity of species, genotypes, soils and climates, different agronomic practices are being implemented and tested in different regions, including Denmark, Turkey, Bulgaria, Chile and Canada. Whereas Chilean started their trials by developing a nursery built on the "Tunnel" greenhouse model with a capacity to accommodate 15.000 cuttings, under an irrigation system with nebulizers to reduce temperature and humidification before a devel‐ opmental stage in the fields, the Danish, Swedish and Canadian choose to established field trials using wild cutting, spacing, density and nutrient management trials [28, 55]. In Sweden, the germplasm used were mostly concentrated on the Scandinavian *Rosa* species of section *Caninae* especially, *R. dumalis, R. rubiginosa* and their interspecific hybrids [86] whereas Danish rosehips are produced mainly from *R. canina* (www.hyben-vital.com) although it may also involve other Scandinavian species. In Chile, the current production is mainly focused on wild hand-harvested hips from uncharacterized and naturalized species introduced to south America by Spanish and is mostly a mixture of *R. rubiginosa, R. canina, R. moschata* and many other species found in western Europe [66]. In Prince Edwards Island, (Canada), current recent genetic study based on 30 wild ecotypes collected from this province suggested that all accessions currently under field trial are from *R. virginiana* and its natural hybrids with *R. Carolina* (Ghose et al, submitted). At present, very few cultivars have been named and released for commercial fruit production. One cultivar, the cultivar "Mechthilde von Neuerburg" derived from *R. rubiginosa* was reported in Germany. Two cultivars (Sylwia and Sylwana) derived from *R. canina* were reported in Poland, whereas cultivar Plovdiv 1 from *R. canina*, and cultivar Karpatia from *R. villosa* were reported in Bulgaria and Slovakia, respectively [13]. For all of these semi-domesticated wild rosehips, it is not known or reported whether the ongoing domestication process has already impacted on some of the phenotypic traits such fruit size, fruit setting or metabolite profile. By comparing the pomology characteristics of 5 wild rosehip ecotypes growing in the wild or in the field settings, we observed that the field setting contributed to increase the fruits size and delayed the maturity when compared with growing in the wild, suggesting an occurrence of a domestication syndrome for these traits (Fofana, personal observation). However, no significant difference was found between the two

48 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

environments for the number of seed in each of the ecotype.

Although originally native to temperate regions of the globe, roses have adapted to warmer regions and grow well now in very diversified habitats and soil types [13, 79]. The soil should be well drained though and not heavy. Species preference for soil type has nonetheless been reported. *R. villosa* was reported to grow better in a dry soil with low calcium content whereas *R. canina* and *R. dumalis* prefer more calcareous soil. *R. rubiginosa* also prefers more calcium and grows well in a relatively heavy soil [13]. *R. palustris* grows in marshes and *R. nitida* in bogs. Similarly, *R. virginiana* likes salt marshes and salty soils (Joly, personal communications).

*3.3.2.1.1. Soils and climates*

Barry et al [55] described the first time the establishment of field trial for North American wild roses belonging to the *R. carolina* complex, with as an objective to investigate the effects of several field management practices on commercial rosehip production in Atlantic Canada. Treatments were applied at planting in a factorial randomized complete block design in June 2004 and included three in-row mulch (none, bark, and straw) treatments, three in-row fertility (none, compost, and fertilizer) treatments, and two interrow management (tilled and sod) treatments. The compost consisted of an initial mix of softwood sawdust, lobster waste, and old hay. Prior to planting, compost was applied at 60 t ha−1 (54 kg plot−1) in a 1-m band over the row and was incorporated by hand raking. The fertilizer used was a commercial grade (5N-20P-20K). This fertilizer formulation was chosen for use during the first year to promote root development and plant establishment. During the second year (2005), compost was reapplied as top-dress on 22 June 2005 and the fertilizer used was a commercial grade (10N-10P-10K), which was applied as top-dress on 25 May 2005. A fertilizer with higher nitrogen content was chosen with the aim of improving overall plant health and yield during the second growing season. Fertilizer was applied at a rate of 800 kg ha−1 (648 g plot−1) in a 1-m band over the planting row. In Dogroses, Werlemark and Nybom [13] reported 50 g NPK for each plant at planting and 300 kg/ha of organic-mineral NKP in the subsequent year, with additional calcium amendment depending on soil types and species. In Prince Edwards Island, mulching increased nutrient uptake of N and P and increased plant growth. Fertilizer increased plant growth and yield of rose hips compared to no fertilizer or compost treatments. Tilled interrow treatment increased in shoot lengths, diameters, and plant spreads compared to interrow sod. The study indicated that during the early establishment years of a rose hip plantation in Atlantic Canada, wild roses grow best with the use of mulch, fertilizer, and tillage between the rows [55].

#### *3.3.2.1.3. Pests and diseases management*

Traditionally, fungal diseases such as black spot caused by *Diplocarpon rosae*, powdery mildew (*Podosphaera pannosa*) rusts (*Phargmidium spp*) and leaf spot *(Sphaceloma rosarum)* have been reported to be problematic in ornamental roses [87-90] and field-grown dogroses [13, 48, 91, 92]. These fungal diseases management is carried through fungicide treatment [93] and selection of genetic resistance [94-96]. Genetic resistance sources within wild rose species within *Caninae* section have been investigated for field rosehip production. Fungal disease tolerance characteristics were identified in *R. rubiginosa* and in interspecific hybrids involving species from *Caninae* and *Cinnamomae* sections [48]. Up to date, no such disease resistance screening has been performed within the *R. carolina* complex for a commercial wild rosehips production in North American. However, our observations in the field showed evidence of these diseases on PEI wild roses (Figure 5). Research in this field should be carried to mitigate the disease incidence in their new field environment. As for any crop, introduction of elite genotypes in cropping systems for rosehips production will lead to a decreased genetic diversity of the cultigens. It is thus anticipated that more susceptibility to major diseases could be observed in the field as compared to the wild populations from which they derive. The preservation of natural habitats hosting the wild populations is of great importance to ensure an availability of genetic stocks to be used in the introgression of disease resistance genes from the wild types to the cultigens.

Insects such aphids (*Aphidina*), grasshoppers (*Orthoptera*), mites (*Tetranychidae*), sawflies (*Tenthredinidae*), gall-making cynipids (*Diplolepis*) as well as the rosehip fly (*Rhagoletis alter‐ nate*) have also been reported in dogrose orchards and to cause severe damage in some cases [13]. Nematode (*Pratylenchus penetrans*) is causal pests of severe lesions to roots in a wide range of ornamental hosts, including roses, mainly in temperate regions. Peng [97] reported that *R. virginiana* is a good nematode resistance source. Because Prince Edwards Island is world leading potato producing area with prevalence of nematodes in the agricultural landscape, development of rosehips orchards with *R. virginiana* genetic background could be a mean for reducing nematode populations in highly infested fields.

**Figure 5.** Foliar and fruits diseases in wild roses. A, powdery mildew; B, leaf spot; C, lesions on immature rosehips probably caused by *Phargmidium spp* (Rust) or *Sphaceloma rosarum* (leaf spot).

#### *3.3.2.1.4. Yield and storage*

(none, compost, and fertilizer) treatments, and two interrow management (tilled and sod) treatments. The compost consisted of an initial mix of softwood sawdust, lobster waste, and old hay. Prior to planting, compost was applied at 60 t ha−1 (54 kg plot−1) in a 1-m band over the row and was incorporated by hand raking. The fertilizer used was a commercial grade (5N-20P-20K). This fertilizer formulation was chosen for use during the first year to promote root development and plant establishment. During the second year (2005), compost was reapplied as top-dress on 22 June 2005 and the fertilizer used was a commercial grade (10N-10P-10K), which was applied as top-dress on 25 May 2005. A fertilizer with higher nitrogen content was chosen with the aim of improving overall plant health and yield during the second growing season. Fertilizer was applied at a rate of 800 kg ha−1 (648 g plot−1) in a 1-m band over the planting row. In Dogroses, Werlemark and Nybom [13] reported 50 g NPK for each plant at planting and 300 kg/ha of organic-mineral NKP in the subsequent year, with additional calcium amendment depending on soil types and species. In Prince Edwards Island, mulching increased nutrient uptake of N and P and increased plant growth. Fertilizer increased plant growth and yield of rose hips compared to no fertilizer or compost treatments. Tilled interrow treatment increased in shoot lengths, diameters, and plant spreads compared to interrow sod. The study indicated that during the early establishment years of a rose hip plantation in Atlantic Canada, wild roses grow best with the use of mulch, fertilizer, and tillage

50 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

Traditionally, fungal diseases such as black spot caused by *Diplocarpon rosae*, powdery mildew (*Podosphaera pannosa*) rusts (*Phargmidium spp*) and leaf spot *(Sphaceloma rosarum)* have been reported to be problematic in ornamental roses [87-90] and field-grown dogroses [13, 48, 91, 92]. These fungal diseases management is carried through fungicide treatment [93] and selection of genetic resistance [94-96]. Genetic resistance sources within wild rose species within *Caninae* section have been investigated for field rosehip production. Fungal disease tolerance characteristics were identified in *R. rubiginosa* and in interspecific hybrids involving species from *Caninae* and *Cinnamomae* sections [48]. Up to date, no such disease resistance screening has been performed within the *R. carolina* complex for a commercial wild rosehips production in North American. However, our observations in the field showed evidence of these diseases on PEI wild roses (Figure 5). Research in this field should be carried to mitigate the disease incidence in their new field environment. As for any crop, introduction of elite genotypes in cropping systems for rosehips production will lead to a decreased genetic diversity of the cultigens. It is thus anticipated that more susceptibility to major diseases could be observed in the field as compared to the wild populations from which they derive. The preservation of natural habitats hosting the wild populations is of great importance to ensure an availability of genetic stocks to be used in the introgression of disease resistance genes from

Insects such aphids (*Aphidina*), grasshoppers (*Orthoptera*), mites (*Tetranychidae*), sawflies (*Tenthredinidae*), gall-making cynipids (*Diplolepis*) as well as the rosehip fly (*Rhagoletis alter‐ nate*) have also been reported in dogrose orchards and to cause severe damage in some cases

between the rows [55].

*3.3.2.1.3. Pests and diseases management*

the wild types to the cultigens.

Rosehip yield vary considerably depending on the plant material, cultivation procedures, age of orchard, and harvesting methods. Werlemark and Nybom [13] reported that up to 8 kg of rosehips per bush could be harvested by hand in commercial planting of dogrose hybrid PiRo 3. Similarly up to 3 t/ha could be obtained from *R. dumalis* and *R. rubiginosa* with mechanical harvesting in Sweden. In these cases however, no mention is made about the age of the orchards as yield increases markedly several years after planting. In contrast, Sanderson and Fillmore [56], reported in 14 rosehip ecotypes of the *R. Carolina* complex grown in field condition an average rosehip yield ranging between 411 and 2000 kg/ha, with a fruit mean weight of 1.01 – 1.62 g, over the first four hand harvesting years. The lowest and highest yielding selections showed 910 and 3634 kg/ha in the fourth years, respectively (Table 3).

Compared with reports by Ercisli and Guleryuz [98], Dogan and Kazankaya [99], Güneş and Dölek [100], the fruit weight reported by Sanderson is lower but showed relatively narrow range of variation between ecotypes, reflecting the relatively narrow genetic diversity among these ecotypes. Joly (personal communication) reported that *R. virginiana* and *R. Arkansana* are the two species with the greatest number of fruits per flowering branches. They have more fruits than *R. carolina* and the height of *R. virginiana* makes it one of the most productive North American roses. To preserve the integrity of rosehip bioactives, the postharvest handling and storage conditions are key factors. Both sun-drying and mechanical dryers are being used at commercial scale and the reader can see more details in Werlemark and Nybom [13].


**Table 3.** Yield progression over four years after plantation and mean fruit weight of 14 rosehip ecotypes grown in field (2006-2009)

#### *3.3.2.2. Biotechnology*

One of the shortcoming issues for the establishment of commercial rosehip production orchard is the availability plant materials for large acreages. So far, all established fields are based on cuttings or seedlings obtained from wild selections. Because of the genetic diversity within the genus *Rosa* and morphological similarities between species, hybrids (interspecific and intraspecific) and their parental species at the collection sites, an accurate identification at the collection site and the traceability of the putative cultivars under development is challenging and not guaranteed. This issue will become major issues in a near future as rosehip prove‐ nances will increase and the bioactive metabolites that are associated to each species, prove‐ nance, and ecotypes are made available for marketing purposes. Thus, the use of combined morphological, cytological, and molecular biology tools for assigning a genetic identity, and the use of regeneration technologies that ensure mass plant production and ensuring the genetic integrity of clones is a research direction that should be undertaken similarly to the ornamental flower industry.

#### *3.3.2.2.1. Regeneration and propagation*

#### *3.3.2.2.1.1. Regeneration by seed*

Compared with reports by Ercisli and Guleryuz [98], Dogan and Kazankaya [99], Güneş and Dölek [100], the fruit weight reported by Sanderson is lower but showed relatively narrow range of variation between ecotypes, reflecting the relatively narrow genetic diversity among these ecotypes. Joly (personal communication) reported that *R. virginiana* and *R. Arkansana* are the two species with the greatest number of fruits per flowering branches. They have more fruits than *R. carolina* and the height of *R. virginiana* makes it one of the most productive North American roses. To preserve the integrity of rosehip bioactives, the postharvest handling and storage conditions are key factors. Both sun-drying and mechanical dryers are being used at

52 Using Old Solutions to New Problems - Natural Drug Discovery in the 21st Century

commercial scale and the reader can see more details in Werlemark and Nybom [13].

s26 877 1413 2368 3634 2000 1.62 s30 347 569 1557 2676 1431 1.31 s28 498 335 1136 1759 946 1.57 s22 416 422 831 1464 783 1.29 s67 270 338 910 1440 740 1.39 s25 355 116 562 1725 719 1.29 s57 195 371 941 1178 675 1.03 s33 395 330 654 1227 657 1.33 s55 313 384 679 1167 638 1.42 s36 181 166 862 1307 622 1.01 s140 300 186 576 1342 610 1.21 s142 406 430 464 956 568 1.17 s68 284 281 430 1092 514 1.21 s122 246 83 416 910 411 1.12 Grand mean 363 387 885 1563 808 1.28

**Table 3.** Yield progression over four years after plantation and mean fruit weight of 14 rosehip ecotypes grown in

One of the shortcoming issues for the establishment of commercial rosehip production orchard is the availability plant materials for large acreages. So far, all established fields are based on cuttings or seedlings obtained from wild selections. Because of the genetic diversity within the genus *Rosa* and morphological similarities between species, hybrids (interspecific and intraspecific) and their parental species at the collection sites, an accurate identification at the collection site and the traceability of the putative cultivars under development is challenging and not guaranteed. This issue will become major issues in a near future as rosehip prove‐ nances will increase and the bioactive metabolites that are associated to each species, prove‐ nance, and ecotypes are made available for marketing purposes. Thus, the use of combined

**Biological yield Mean fruit**

**(kg ha-1) (g)**

**2006 2007 2008 2009 Mean weight**

**Selection**

field (2006-2009)

*3.3.2.2. Biotechnology*

The use of plant regeneration from seed for commercial production has been reported [85, 101]. It ensures the production of higher number of plants for field planting in a relatively short period of time. However, the mating system of *Rosa* species is a major source of genetic variability between plant materials obtained using such an approach, especially when the seed is collected from uncontrolled sources like wild plants.

#### *3.3.2.2.1.2. Cutting and explants*

Cuttings and explants are currently the materials of choice in commercial wild rose production [64, 86, 101], and most, if not all, of these explants (Figure 6) are derived from wild plants. Wild rose plants grow in the nature as populations that can involve different species, interspecific and intraspecifc hybrids, parental and sibling all growing in a confined area. Collecting cuttings in such an environment, even from the same patch, does not ensure the genetic integrity of the collected material for propagation. Once collected, the material should be well characterized and identified. Now, remains our ability to get enough characterized plant materials for large field planting. We believe that the well characterized plant material should be used as starting point for plant regeneration and mass production in the form of rooted seedling or cuttings. This is the approach we pursue in Canada for commercial wild rose production (Figure 7).

#### *3.3.2.2.1.3. Tissue culture*

Tissue protocols have been developed and available for flower roses [102-104] and could be applied to rosehip production. Once elite genotypes such as those reported by Sanderson and Fillmore [56] are identified, tissue culture should be able to ensure a sustainable plant pro‐ duction or field planting by growers (Figure 7).

#### *3.3.2.2.2. Cell culture*

Similar to tissue culture, rose plants can be regenerated by cell culture. Contrary to tis‐ sue culture however, the new plants are obtained from callus generated from sterile ex‐ plants. This method leads to pure line but can also create new lines different from the mother plant from which the explant was obtained because of somaclonal variations that may occur during the induction of callus and regeneration processes. Thus, for the pro‐ duction of mass plant production from a selected elite wild ecotype, tissue culture ap‐ pears more appropriate as it minimizes the risk of somaclonal variations while showing high rate of plant multiplication.

**Figure 6.** Rose cuttings for multiplication. Sterile rose dormant stems were conditioned to break dormancy. Note the active buds sprouting.

**Figure 7.** Mass rosehip plant regeneration from active buds of well characterized rosehip genotypes. A, active buds in re‐ generation media; B, regenerated rose plant; C, plant multiplication in rooting media; and D, acclimation in greenhouse.
