A review of genetic resources of almonds and stone fruits (<Emphasis Type="Italic">Prunus</Emphasis> spp.) in Iran

Iran is amongst the countries in the world widely known for cultivation of Prunus spp. (or stone fruits). It is both a centre of origin and diversity of the stone fruits. Numerous wild species as well as many cultivars and landraces of these fruit crops are important genetic resources today in Iran and can be used for improvement and breeding of scion and rootstock cultivars which are resistant to many biotic and abiotic stresses through modern genomics and genetic technologies. This paper discusses the distribution, ethno-botany, diversity and utilization of wild and domesticated genetic resources of stone fruits including almond (Prunus dulcis (Miller) D. A. Webb.), peach and nectarine (P. persica Batsch), European and Japanese plum (P. × domestica L., and P. salicina L., respectively), sweet and sour cherry (P. avium L., and P. cerasus L., respectively), and apricot (P. armeniaca L.), all of which are members of the Rosaceae family. The goal of this paper is to highlight the importance of Iran as a main contributor to the diversity of Prunus genetic resources in the world, as well as, present major achievements regarding identification, collection, evaluation, conservation and utilization of this valuable genetic resource in Iran.     

Agro-morphological and molecular variability in <Emphasis Type="Italic">Triticum boeoticum</Emphasis> accessions from Zagros Mountains,Iran

Genetic erosion in cultivated wheat provides a good reason for studying genetic diversity in crop wild relatives. In the present study, genetic diversity of 32 accessions belonging to T. boeoticum species collected from different parts of Iran were evaluated using 13 morphological traits as well as ten inter-simple sequence repeat primers. Statistical analysis for morphological traits showed significant differences among accessions (except number of fertile tillers and total tillers per plant). In principal component analysis, the first three PCA showed 82.65 % of the total morphological variation. Based on the morphological traits, accessions were separated into two main groups by cluster analysis. In molecular analysis, polymerase chain reactions amplified 105 DNA fragments, out of which, 95 (90.47 %) were polymorphic. From geographic perspective, the accessions sampled from western and southwestern of Iran showed the highest and lowest polymorphism, respectively. However, the maximum values of effective number of alleles (Ne), Nei’s gene diversity (He) and Shannon’s information index (I) was related to accessions collected from NW regions. Also, according to cluster analysis and PCoA plot genetic diversity was not related to geographical distribution. Overall, our results revealed a remarkable level of genetic diversity among studied Iranian T. boeoticum accessions; especially accessions collected from Kermanshah and Lorestan provinces, which can be of interest for future breeding programs. So, conservation of germplasm of these areas is recommended.     

Retrotransposon-based sequence-specific amplified polymorphism markers for the analysis of genetic diversity and phylogeny in <Emphasis Type="Italic">Malus</Emphasis> Mill. (Rosaceae)

Sequence-Specific Amplified Polymorphism (S-SAP)-analysis of 131 accessions of different species from five sections of the genus Malus was carried out to study genetic diversity, and to clarify phylogenetic and taxonomic issues. S-SAP-markers, based on long terminal repeat (LTR)-retrotransposons TRIM2 and dem1, were developed, which identified 679 polymorphic fragments in the studied Malus species. S-SAP technique proved to be effective for taxonomic studies in Malus. The obtained results generally support the existing sectional taxonomy in the genus Malus and allowed to determine the taxonomic status of some Russian landraces. The genetic diversity and taxonomic status of the Russian apple Antonovka landraces, widely used in breeding programs for their increased adaptation to abiotic stress and scab resistance, were clarified. All of them belong to M. domestica, section Malus. However, Antonovka Olginskaya might have a hybrid origin with some contribution from Gymnomeles species according to PCO analysis data. The taxonomic status was resolved for another Russian landrace, Yakutskaya, which exhibits increased winter hardiness and drought resistance; it belongs to section Gymnomeles, with a high resemblance to M. baccata.     

Morphological characterization of snake melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">flexuosus</Emphasis>) populations from Palestine

Genetic diversity in 50 snake melon accessions collected from Palestine (West Bank) was assessed by examining variation in 17 phenotypic characters. These accessions belonged to four important landraces of Cucumis melo var. flexuosus: Green “Baladi” (GB), white Baladi (WB), green Sahouri (GS), and white Sahouri (WS). Principal component analysis (PCA) and a dendrogram were performed to determine relationships among populations and to obtain information on the usefulness of those characters for the definition of cultivars. PCA revealed that secondary fruit skin color, flesh color, primary fruit skin color, and secondary skin color pattern were the principal characters to discriminate melon accessions examined in the present study. According to the scatter diagram and dendrogram, landraces of C. melo var. flexuosus: GB, WB, GS, and WS formed different clusters. However, based on Euclidean genetic coefficient distances, GB and WB had the least degree of relatedness with GS and WS, indicating distantly related landraces (Baladi and Sahouri). On the other hand, the highest degree of relatedness was detected between WS on one hand, and both GS and WB on the other indicating closely related cultivars. Fruit traits variability among the different snake melon landraces was evaluated and discussed in this study. This evaluation of fruit trait variability can assist geneticists and breeders to identify populations with desirable characteristics for inclusion in cultivars breeding programs.     

Genetic structure of Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L.) landraces grown in the Mayan area

Lima bean (Phaseolus lunatus L.) is an important crop in the Mayan culture. The Mayan area, considered as a main center of Mesoamerican diversity, has been divided into two subareas: the Mayan lowlands and the Mayan highlands. The Yucatan Peninsula is part of the Mayan lowlands and holds the highest number of Lima bean landraces of Mexico, but Lima beans are in high risk of genetic erosion due to intensification of the traditional Mayan agriculture. However, information on genetic diversity of Lima beans of the Mayan highlands is lacking. By using 46 landraces collected in the Mayan area (23 from each subarea) and 73 ISSR loci (inter-simple sequence repeats), we analyzed the structure, diversity and genetic relationships of Lima beans of this part of Mesoamerica. High levels of diversity (H _BAY = 0.45) and genetic structure (F _ST = 0.66) were found for the whole Mayan area. Genetic diversity in the Mayan lowlands was apparently higher than the Mayan highlands (H _BAY = 0.44 and 0.36, respectively); but differences were not statistically significant. Genetic structure between the subareas was high (AMOVA = 30% of total variation), most landraces grouping according to their geographic origin. This study shows the importance of the Mayan culture in the diversification and conservation of Lima beans. The results provide important information that should be considered when implementing strategies to collect Lima bean landraces and planning in situ and ex situ programs to conserve these landraces in the Mayan region.     

Population genetics of traditional landraces of <Emphasis Type="Italic">Cucurbita pepo</Emphasis> L., 1753 in the cloud forest in Baja Verapaz,Guatemala

Little is known regarding the effect of fragmentation and human agricultural management on the genetic variation and gene flow of Cucurbita pepo L., 1753 in moderate fragmented areas in central Guatemala. We hypothesize that the genetic variation of C. pepo is affected by forest fragmentation and by traditional agricultural management. Therefore, we aim to determine: (1) the genetic diversity and genetic structure of C. pepo in the Cloud Forest Corridor (CFC) (2) the extent of genetic admixture between commercial variety (CV) and traditional landraces (TL) of C. pepo, (3) the effect of habitat fragmentation in the population genetics of C. pepo with a landscape approach, and (4) the potential relationship between traditional management practices and genetic diversity of C. pepo in the CFC. We detected the existence of high level of genetic diversity (AR = 3.43; He = 0.50), inbreeding (Fis = 0.25) and moderate population structure of C. pepo in the CFC (Fst = 0.16). No correlation between landscape and genetics was found. Also, we found high genetic admixture between CV and TL. It seems that human practices, mainly related with seed exchange patterns, could affect genetic diversity of C. pepo in the CFC. C. pepo populations in the CFC are structured, with inbreeding, and show admixture with the CV, an aspect that could affect its genetic diversity. The agricultural management influenced the population genetics of C. pepo in the CFC, but the landscape did not. We suggest that special efforts should be made to preserve the diversity of this important indigenous food source for Guatemalan people as well as their management practices.     

Genetic diversity and relationships of ancient Chinese fir (<Emphasis Type="Italic">Cunninghamia lanceolata</Emphasis>) genotypes revealed by sequence-related amplified polymorphism markers

Some more than a century old Chinese fir genotypes with high biomass that are now rare in China were historically called “the king of Chinese fir”. The genetic diversity and relationships of those ancient Chinese fir genotypes were investigated using morphological analysis and sequence-related amplified polymorphism markers. The morphological analysis indicated that the tree volume parameter had the maximum variable coefficient value (70.2 %). In total, 18 selected primers generated 154 bands, and 150 bands were polymorphic (97.4 %). Higher values of genetic diversity parameters (h = 0.3593, I = 0.5283) were maintained at the species level, indicating that the ancient Chinese fir in China has retained a relatively high level of genetic diversity. AMOVA indicated that 73.16 % of the variation resided within provenances. The UPGMA dendrogram and genetic structure analysis identified 3 major clusters and grouped the genotypes in agreement with their geographic origins. A Mantel test revealed a significant correlation between genetic distances and geographic distances among the genotypes (r = 0.4275, p < 0.01). Overall, we recommend a combination of conservation measures including a germplasm repository and the implementation of in situ conservation.     

Biodiversity in Ethiopian linseed (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.): molecular characterization of landraces and some wild species

Molecular characterization of germplasm is important for sustainable exploitation of crops. DNA diversity was measured using inter-retrotransposon-amplified-polymorphism and inter-simple-sequence-repeat markers in 203 Ethiopian landraces and reference varieties of linseed (flax, Linum usitatissimum) and wild Linum species. Molecular diversity was high (PIC, 0.16; GD, 0.19) compared to other reports from the species. Genotyping separated reference from landrace accessions, and clustered landrace accessions from different altitudes and geographical regions. Collections showed evidence for recent introduction of varieties in some regions. The phylogeny supported L. bienne Mill. as the progenitor of domesticated L. usitatissimum. Markers developed here will be useful for genetic mapping and selection of breeding lines. The results show the range of characters that can be exploited in breeding lines appropriate for smallholder and commercial farmers in Ethiopia, producing a sustainable, secure, high-value crop meeting agricultural, economic and cultural needs.     

First insight into genetic diversity and population structure of the Caucasian wild apple (<Emphasis Type="Italic">Malus orientalis</Emphasis> Uglitzk.) in the Hyrcanian forest (Iran) and its resistance to apple scab and powdery mildew

Investigating population genetic structure and diversity, and resistance to pathogens in crop wild relatives are key steps to assess appropriate conservation and breeding programs. The Caucasian wild apple (Malus orientalis Uglitzk.) is an emblematic fruit tree of the Hyrcanian forest and is supposed to be a contributor to the cultivated apple genome (Malus domestica Borkh. Yet, no study has investigated its population structure, diversity and susceptibility to the two main pathogens of apples, the apple scab (Venturia inaqualis) and the powdery mildew (Podosphaera leucotricha). Here, we investigated population genetic structure and diversity of M. orientalis in the Hyrcanian forest as a mean to identifying future targeted populations for apple conservation and breeding programs. We genotyped using multilocus microsatellite 100 M. orientalis trees sampled in 14 sites. These trees were also screened for presence/absence of six (Rvi6, Vr, Rvi4, Rvi15, Rvi5 and Rvi11) and three (Pl-1, Pl-w, Pl-d) resistance genes to the apple scab and the powdery mildew respectively. Our results showed significant but weak between-site genetic differentiation and isolation by distance pattern suggesting substantial historical gene flow for M. orientalis in this area. We also detected a West-Eastern genetic structure across the Hyrcanian forest with five main populations showing admixture. We also showed a high diversity of resistance genes to apple scab across sites; in contrast, we only found one resistance gene to powdery mildew. These results are a first glimpse to settle wild apple conservation programs in Iran and pinpoint Iranian wild apple populations as an untapped source for apple breeding.     

Genetic diversity,genetic structure and migration routes of wild <Emphasis Type="Italic">Brassica juncea</Emphasis> in China assessed by SSR markers

The wild Brassica juncea (L.) Czern. et Coss., systematically belonging to the genus Brassica L. in the Cruciferae family, has become a noxious weed for cropping systems nowadays. Here, simple sequence repeat (SSR) markers were applied to investigate the genetic diversity, genetic structure and migration routes of the wild B. juncea populations in China. The results showed that a total of 90 alleles, with extensive allelic diversity, were observed at the 11 SSR loci of the wild B. juncea. The STRUCTURE analysis indicated that all the 25 wild populations were best described as belonging to two lineages. High Fst value (0.568), together with the partitioning, provided significant evidence for lineage differentiation in wild B. juncea. The high differentiation between the two lineages was, perhaps, due to limited gene flow (Nm?=?0.301) of this species. The analysis of molecular variance with distances among individuals corrected for the dominant nature of SSRs showed that most of the variation (59%) occurred within populations, and the remaining 41% variance was attributed to differences among populations. The distribution of diversity across China was significantly geographically dependent. NJ cluster analysis, based on genetic distance, grouped populations geographically, which further corroborated spatial pattern of two lineages. Based on these results, two routes were proposed for the migration of wild B. juncea in China after its origin from northwest China, heading east along the Yellow River or Yangtze River, respectively. We concluded that China, especially the northwest, is one of the primary origins of B. juncea.     

Remarks on genetic diversity and relationship of <Emphasis Type="Italic">Punica protopunica</Emphasis> and <Emphasis Type="Italic">P. granatum</Emphasis> assessed by molecular analyses

Here, two Punica species, viz., P. protopunica Balf. fil., reported as native to Socotra, and P. granatum L., were compared for the first time. Analysis of one P. protopunica and eleven P. granatum accessions was performed using three molecular markers, i.e., sequence related amplified polymorphism (SRAP), target region amplification polymorphism (TRAP), and intron targeted amplified polymorphism (ITAP), along with analysis of pgWD40 sequences, a gene involved in anthocyanin biosynthesis. All markers revealed the relationship between the two species and placed them at 33% similarity. SRAP, TRAP, and ITAP generated a total of 299, 260, and 160 bands, respectively. Of these, 78, 74, and 41 bands were specific for P. protopunica, and 92, 85, and 57 bands, respectively, were shared between both species. Sequence analysis of pgWD40~870 bp amplicons exhibited 100% identity among P. granatum accessions and 98% identity to that of P. protopunica. Phylogenetic analysis of WD40 sequences from monocot and dicot species, including both Punica species confirmed the relation between P. protopunica and P. granatum, supporting earlier reports that P. protopunica could be an ancestral species of P. granatum. Furthermore, the genetic diversity among and within P. granatum accessions from Egypt (3), Mexico (5), and Yemen (3) was assessed. Molecular marker-based relationships among region-bulked accessions was approximately the same (~90% similarity), whereas the degree of genetic variation was altered within each region. Specific bands (alleles) for accessions of each region along with those shared among them were identified. Thus, these bands could be used for pomegranate genotyping and breeding programs.     

Evaluation of <Emphasis Type="Italic">Rubus</Emphasis> genetic resources on their resistance to cane disease

Raspberry cane disease, caused by a complex of fungi, is amongst the most devastating problems for raspberry production. Using resistant or highly tolerant genotypes is a promising strategy. However, until now, cultivars with good field resistance for German fruit growers as well as the primary causal pathogens of cane disease present in the eastern part of Germany, Saxony state are still unknown. The primary objective of this study was to identify the primary disease causing organisms and resistant Rubus germplasm for future breeding efforts. From 2012 to 2014, we evaluated the degree of susceptibility to cane diseases on 213 raspberry cultivars at two different locations using a rating scale. We also identified the main fungi growing on infected canes. Our results suggest that Fusarium avenaceum is the main fungus causing cane disease in Saxony. The Rubus hybrid cultivars ‘Tayberry’ (2n?=?6x?=?42), ‘Buckingham Tayberry’ (2n?=?6x?=?42) and ‘Dorman Red’ (2n?=?2x?=?14) displayed the highest levels of field resistance at both locations. Moreover, we established an in vitro assay for resistance evaluation which correlates significantly with field data and is particularly suitable for quick assessment of resistance against F. avenaceum in breeding material. Future breeding programs, aiming at cultivars with enhanced resistance towards cane diseases can use the presented data for parental selection and may employ our in vitro F. avenaceum inoculation method for parental testing and progeny selection.     

Farmers perception of pesticide use and genetic erosion of landraces of tetraploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) in Ethiopia

Perception of farmers’ about the use of pesticides and genetic erosion of tetraploid wheat landraces of Ethiopia was assessed through focus group discussions with farmers, on-farm observations, personal interviews with farmers, by using structured questionnaires of temporal and spatial methods. A total of 1496 farmers from seven provinces in the country were interviewed. Farmers’ knowledge about pesticide increases suggests that they are not happy on using chemicals because of their negative impact on farm land. About 75 % of the farmers believe that, although the use of pesticides may increase the production of wheat, it has a negative impact on (human) health and environment. Women showed a higher concern for pesticides’ harmfulness than men. Farmers’ valuation of genetic erosion was estimated as reduced importance of landraces, as shown by a the lower proportion of landraces either grown or sold on the market. The four most important factors cited for loss of landraces were reduction in land area per capita, displacement by released/modern varieties of hexaploid wheat and teff, reduced benefit from landraces, and displacement by other crops and chat. Genetic erosion of 100 % was observed for Triticum dicoccon in the provinces of Gojam and Gonder and for T. polonicum in Tigray and Gojam. Overall, genetic erosion in the country was 32.0, 35.3, 55.9, 84.4 and 84.4 % for T. durum Desf., T. turgidum L., T. aethiopicum Jakubz., T. polonicum L. and T. dicoccon Schrank, respectively.     

<Emphasis Type="Italic">Panax</Emphasis> species identification with the assistance of DNA data

Panax species have been used as medicinal plants in China and adjacent areas, but the difficulty in wild species identification based on morphological characters prevents the exploration and utilization of these resources. To assist in identifying the species, three plastid intergenic spacers and four nuclear genes of 129 samples from 20 different localities were sequenced to conduct the clustering analyses. P. stipuleanatus and P. notoginseng were relatively easy to be distinguished from other species based on morphological characters and clustering results. Three samples collected from Laos grouped together with those collected from Jinping (China), and they were all identified as P. vietnamensis. However, the fourth one LW4 had closer genetic relationship to P. notoginseng than to P. vietnamensis. Although they were morphologically different in petiolule length, the samples preliminarily identified as P. zingiberensis grouped together in the clustering analyses, and the results of STRUCTURE analysis suggested that samples collected from Zhenyuan shared some genetic component with P. vietnamensis. As for the samples identified as P. japonicus and P. wangianus, there were so many variations in leaves, rhizomes and rootstocks that it was difficult to differentiate them from each other. Molecular data also showed that these taxa were genetically admixed and the samples identified as P. wangianus could be classified into P. japonicus. The nucleotide diversity results indicated that the wild relatives had more substantial genetic diversity compared to P. notoginseng.     

Ecological,morphological and genetic diversity in <Emphasis Type="Italic">Oryza rufipogon</Emphasis> Griff. (Poaceae) from Hainan Island,China

In this study, ecological, morphological and genetic diversity in 11 surviving populations of Oryza rufipogon Griff. from Hainan Island, China were investigated. These data served to provide much needed data in support of conservation strategies. The results divided O. rufipogon into six ecotypes. The cluster analysis based on 19 morphological features showed clear differentiation among populations or within populations. The cluster analysis based on SSR also revealed similar results. The ecological, morphological and genetic diversity observed in O. rufipogon resulted in the following conservation recommendations: (1) the four currently protected O. rufipogon localities should be maintained, due to the different ecological communities represented at these sites; (2) some populations, such as HL (Hele county, Hainan Island), should be protected, but some populations, such as FC (Fucheng county, Hainan Island) and DL (Dalu county, Hainan Island), might not be protected. (3) An isolation zone should be established to avoid possible introgression between O. sativa L. and O. rufipogon; and human activities that impact O. rufipogon should be reduced or eliminated.     

Morphological and molecular characterization revealed high species level diversity among cultivated,introduced and wild roses (<Emphasis Type="Italic">Rosa</Emphasis> sp.) of western Himalayan region

High level of genetic diversity and morphological variations were observed among and within the different Rosa species studied. The key traits which differentiated the species were nature of plant, habit, shape of prickles, shape of stipules, leaflet shape, type of leaf margin, vestiture of ventral side of leaflet and flower shape, whereas, traits for distinguishing the accessions within a species (as in case of accessions from western Himalayan region) were flower colour, petal number, shape of hip and hip colour. Some of the accessions from western Himalayan region formed out-groups (IHBT-WR-18, IHBT-WR-21, IHBT-WR-31and IHBT-WR-43B) and could not be classified at the species level. High degree of variations among the accessions of R. multiflora group as compared to variations among the accessions of R. moschata, R. brunonii and R. cathayensis, suggests high level of out-breeding in R. multiflora which may contribute to further variations. Desirable variations were observed for flower colour, petal number, lack of prickles, perpetual flowering, large flower diameter and large fruit size which contribute significantly to diversity of the roses and have potential use in breeding programs.     

Genetic diversity and relationships of wild and cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> germplasms based on sequence-related amplified polymorphism (SRAP) markers

The genus Zanthoxylum, belonging to Rutaceae, has a long history of cultivation both for economic and chemical values in China. To effectively conserve and sustainably utilize this genus resource, a study on genetic diversity and relationships of Zanthoxylum germplasms was carried out by employing SRAP markers. We used 16 primer combinations to assess genetic variations and relationships among 175 accessions from eight cultivated provenances, including Shandong, Henan, Shanxi, Shaanxi, Gansu, Sichuan, Guizhou and Yunnan. A total of 145 clear repetitive and intense bands were yielded, and the percentage of polymorphic bands was 100 % for per primer combination, indicating a relatively high diversity among Zanthoxylum germplasms. From a geographic perspective, the highest genetic diversity level was observed within Guizhou provenance (N _a  = 1.97, Ne = 1.52, H = 0.31, I = 0.46) while Henan provenance had the lowest genetic diversity (N _a  = 1.68, Ne = 1.45, H = 0.25, I = 0.37). Based on AMOVA results, the abundant genetic variation was mainly caused by variation of intra-provenances (84.96 %), rather than among provenances (15.038 %). The results indicated low genetic differentiation (G _st  = 0.133) and high gene flow (N _m  = 3.2605) among provenances. The neighbor-joining tree revealed that the 175 accessions could be divided into four groups, and groupings indicated a divergence between the cultivated accessions of Zanthoxylum bungeanum Maxim. and Z. armatum DC. Moreover, three accessions of Z. piperitum DC. var. inerme without prickles introduced from Japan gathered one cluster. Cluster IV is composed of accessions of different geographical origin, including 11 wild species and 10 cultivated accessions of Z. bungeanum. The cluster analysis also reflected a relatively close relationship between the geographical origins and the classification of accessions in cluster I. Structure analysis indicated that collected Zanthoxylum accessions could be divided into two major groups. The information obtained from our research would benefit to make use of Zanthoxylum germplasms and assist the management of a Zanthoxylum germplasms collection.     

Morphological and genetic diversity of Momordica cochinchinenesis (Cucurbitaceae) in Vietnam and Thailand

Momordica cochinchinensis (Cucurbitaceae) is native to South East Asia and is important for its nutritional and medicinal properties. There is little information available on morphological and genetic diversity, crop selection and strategic germplasm collection for M. cochinchinensis. In this study, 42 accessions of M. cochinchinensis collected from Vietnam, Thailand and Australia were analysed for diversity study both at a morphological and molecular level. Both morphological and molecular analyses were in agreement and clustered the accessions based on the country of origin. The markers investigated in this study could be used for future applications with samples from unknown origins or for gene tracking for important cultivars. The Momordica accessions were highly polymorphic based on the markers suggesting high genetic diversity. Therefore the results from this study will be important in future strategic germplasm collection, conservation, horticultural development and commercial propagation of M. cochinchinensis.