Anne Frary currently works at the Department of Molecular Biology and Genetics, Izmir Institute of Technology. Anne does research in Genetics, Biotechnology and Agricultural Plant Science.
Skills and Expertise
Research Items (97)
More than half of European hazelnut (Corylus avellana) production occurs in Turkey. Despite this dominance, the yield of Turkish cultivars has remained stagnant over the past 10 years with Italian yield nearly double that in Turkey. This difference is due to Turkey’s unique cultivation system; hazelnuts are grown in bushy clusters (“ocak” system), not as single trees. Current hazelnut breeding efforts are shifting toward the development of materials for single plant orchards which are much higher yielding; thus, there is a need to explore germplasm for relevant agro-morphological traits and to determine their genetic control. The objectives of this study were to examine data for 44 such traits in 390 hazelnut accessions: 16 cultivars, 232 landraces and 142 wild individuals from nine provinces in Turkey and to map the loci associated with these characteristics using simple sequence repeat markers. Comparison of cultivars, landraces and wild hazelnut accessions revealed the effects of domestication and selection on the crop and indicated that useful alleles for traits such as cropping and reduced alternate bearing may exist in the wild germplasm. A total of 145 quantitative trait loci (QTL) were detected with the largest proportions identified for involucre (26%) and inflorescence (14%) morphology. Several markers co-localized with more than one trait including markers for male catkin abundance which were shared with plant vigor and height. Similarly, markers for female flower abundance co-localized with suckering and alternate bearing. Such markers and their linked QTL should be studied in more detail as they might help breeders select for plant vigor, decreased suckering and increased flower production: traits which will be extremely useful for Turkey’s transition to single plant orchards.
Turkey is a rich source of European hazelnut (Corylus avellana) germplasm with nearly 400 accessions in the national collection. This genetic material encompasses cultivars, landraces and wild genotypes which were characterized for 12 nut and 13 kernel traits over 2 years in the 1990s. Analysis of these attributes revealed both the positive and negative impacts that human selection and breeding have had on hazelnut. Thus, while selection has resulted in larger nuts and kernels, cultivars have fewer nuts per cluster and kernels with larger internal cavities. Breeding has also resulted in a propensity for cultivars to have higher proportions of double kernels and empty nuts, two traits which reduce quality and yield. In addition, it is clear that while selection has successfully increased hazelnut fat content it has not impacted overall flavor, a much more complex trait. The nut and kernel phenotypic data were combined with genotypic data from 406 simple sequence repeat marker alleles for association mapping of the quantitative trait loci (QTL) for the traits. A total of 78 loci were detected in the population with the highest proportions for nut (24%) and kernel (26%) appearance parameters followed by quality (19%), shell thickness (16%) and yield-related (15%) traits. It is hoped that some of the identified QTL will be useful for future breeding of hazelnut for improved nut and kernel yield and quality.
European hazelnut (Corylus avellana) is a diploid tree species and is widely used in confections. Hazelnuts are, to a large part, produced in Turkey with the cultivar “Tombul” widely grown in the Black Sea region. In this work, the “Tombul” genome was partially sequenced by next-generation sequencing technology yielding 29.2% (111.85 Mb) of the ~ 385 Mb (1C). This sequence information was used to develop genetic markers in order to enable differentiation of material before the long maturation process and to facilitate future breeding strategies. A total of 90,142 simple sequence repeats (SSRs) were identified in the contigs giving a frequency of 1 SSR per 1240 nt in the assembly. Mononucleotides were the most abundant SSR marker type (60.9%) followed by di- and trinucleotides. Primer pairs were designed for 75,139 (83.3%) of the SSRs. Fifty SSR primers were applied to 47 hazelnut accessions from nine countries to test their effectiveness and polymorphism. The markers amplified an average of 3.2 fragments. The highest polymorphism information content value was for cavSSR11062 (0.97) and the lowest (0.04) was for cavSSR13386. Two markers were monomorphic: cavSSR12855 and cavSSR13267. Single-copy SSR primers were also assessed for their ability to discriminate 19 Turkish cultivars, and it was found that seven primer pairs (Cav4217, Cav14875, Cav14418, Cav2704, Cav12862, Cav3909, Cav1361) were sufficient for this task. Thus, this study developed new SSR markers for use in hazelnut breeding and genetic studies and also provide a method to distinguish and identify true-type Turkish cultivars.
- Sep 2018
Cotton is an economically important commodity for nearly fifty industries including the textile sector which is largely based on cotton fiber. Identification of markers linked to loci for fiber traits under drought stress may be particularly beneficial because such loci could provide the genetic adaptability needed to produce good fiber under water limitation. In the present study, 177 simple sequence repeat (SSR) markers were used to detect significant quantitative trait loci (QTLs) linked to 11 fiber quality and plant structure traits in a panel of 99 Upland cotton (Gossypium hirsutum L.) genotypes using GLM and MLM analysis. The fiber quality traits, including fiber length (FL), fiber fineness (FF), fiber strength (FS), fiber elasticity (FE), fiber uniformity (FU), spinning conversion index (SCI), earliness (EAR), 1st position boll retention (1st PBR), 2nd position boll retention (2nd PBR), total boll number (TBN) and plant height (PH), were tested under both well-watered and water-limited irrigations in two locations. At both locations, GLM identified a total of 74 and 70 QTLs under well-watered and water limited conditions, respectively, at p ≤ 0.005. MLM detected seven and 23 QTLs under well-watered and water-limited conditions, respectively. Of the identified QTLs, some QTLs were detected in both locations: three for well-watered and two for water-stress conditions. Moreover, a total of 19 QTLs were stable under both watering-regimes. The QTLs identified herein could be useful in the development of cotton cultivars that have adaptability to drought conditions worldwide.
- Aug 2018
Cotton is an economically important commodity for nearly fifty industries including the textile sector which is largely based on cotton fiber. Identification of markers linked to loci for fiber traits under drought stress may be particularly beneficial because such loci could provide the genetic adaptability needed to produce good fiber under water limitation. In the present study, 177 simple sequence repeat (SSR) markers were used to detect significant quantitative trait loci (QTLs) linked to 11 fiber quality and plant structure traits in a panel of 99 Upland cotton (Gossypium hirsutum L.) genotypes using GLM and MLM analysis. The fiber quality traits, including fiber length (FL), fiber fineness (FF), fiber strength (FS), fiber elasticity (FE), fiber uniformity (FU), spinning conversion index (SCI), earliness (EAR), 1st position boll retention (1st PBR), 2nd position boll retention (2nd PBR), total boll number (TBN) and plant height (PH), were tested under both well-watered and water-limited irrigations in two locations. At both locations, GLM identified a total of 74 and 70 QTLs under well-watered and water limited conditions, respectively, at p≤0.005. MLM detected seven and 23 QTLs under well-watered and water-limited conditions, respectively. Of the identified QTLs, some QTLs were detected in both locations: three for well-watered and two for water-stress conditions. Moreover, a total of 19 QTLs were stable under both watering-regimes. The QTLs identified herein could be useful in the development of cotton cultivars that have adaptability to drought conditions worldwide.
Drought causes serious yield losses in cotton production throughout the world. Association mapping allows identification and localization of the genes controlling drought-related traits which will be helpful in cotton breeding. In the present study, genetic diversity analysis and association mapping of yield and drought traits were performed on a panel of 99 upland cotton genotypes using 177 SSR (simple sequence repeat) markers. Yield parameters and drought tolerance-related traits were evaluated for two seasons under two watering regimes: water-stressed and well-watered. The traits included seed cotton yield (SCY), lint yield (LY), lint percentage (LP), water-use efficiency (WUE), yield potential (YP), yield reduction (YR), yield index (YI), drought sensitivity index (DSI), stress tolerance index (STI), harmonic mean (HM), and geometric mean productivity (GMP). The genotypes with the least change in seed cotton yield under drought stress were Zeta 2, Delcerro, Nazilli 87, and DAK 66/3 which were also the most water-use efficient cultivars. The average genetic diversity of the panel was 0.38. The linkage disequilibrium decayed relatively rapidly at 20–30 cM (r² ≥ 0.5). We identified 30 different SSR markers associated with the traits. Fifteen and 23 SSR markers were linked to the traits under well-watered and water-stress conditions, respectively. To our knowledge, most of these quantitative yield and drought tolerance-associated loci were newly identified. The genetic diversity and association mapping results should facilitate the development of drought-tolerant cotton lines with high yield in molecular breeding programs.
Eggplant is a vegetable crop that is grown around the world and can provide significant nutritive benefits thanks to its abundance of vitamins, phenolics and antioxidants. In addition, eggplant has potential pharmaceutical uses that are just now becoming recognized. As compared to other crops in the Solanaceae, few studies have investigated eggplant's metabolic profile. Metabolomics and metabolic profiling are important platforms for assessing the chemical composition of plants and breeders are increasingly concerned about the nutritional and health benefits of crops. In this review, the historical background and classification of eggplant are shortly explained; then the beneficial phytochemicals, antioxidant activity and health effects of eggplant are discussed in detail.
Stanleya pinnata is a perennial herb that can hyperaccumulate selenium. In the present study, genetic diversity of 15 accessions of S. pinnata originating from different locations in the USA was determined using sequence amplified polymorphism (SRAP) markers. A total of 506 alleles were identified among the 15 accessions and 98% of these alleles were polymorphic. Dice dissimilarity coefficient values between accessions varied from 0.14 to 0.87 with an average of 0.53 indicating that the plant material was highly diverse as expected given its self-incompatibility. A dendrogram was constructed to understand the genetic relationships among the accessions based on the molecular marker data. The dendrogram grouped the 15 S. pinnata accessions into three clusters while population structure analysis divided these accessions into five groups. Clustering was correlated with accession origin.
Olive (Olea europaea L.) is one of the most important crops in the Mediterranean region including Palestine and Turkey. In this study, the genetic diversity and population structure of 19 olive cultivars (15 from Palestine and 4 from Turkey) was assessed using simple sequence repeat (SSR) markers. The 14 SSR markers yielded 110 alleles with 91% polymorphism and 7.8 alleles per marker. Genetic dissimilarity ranged from 0.00 to 0.51 with an average of 0.35. Cluster analysis using the Dice coefficient and the neighbor joining algorithm showed that the 19 cultivars grouped into three clusters, with all of the Turkish cultivars in one subcluster. Nabali Baldi from the West Bank and Souri from the Gaza Strip were identical and therefore, synonyms. The highest dissimilarity was between the Turkish Mut and Spanish Arbequina cultivars. In contrast to our expectation, two of the cultivars of supposed Palestinian origin clustered with an Italian cultivar suggesting that it may have originated there. Population structure analysis that assigned the 19 cultivars to two subpopulations.
The aim of this study was to compare the performance of a DNA-barcode assay with fatty acid profile analysis to authenticate the botanical origin of olive oil. To achieve this aim, we performed a PCR-capillary electrophoresis (PCR-CE) approach on olive oil: seed oil blends using the plastid trnL (UAA) intron barcode. In parallel to genomic analysis, we subjected the samples to gas chromatography analysis of fatty acid composition. While the PCR-CE assay proved equally efficient as gas chromatography analysis in detecting adulteration with soybean, palm, rapeseed, sunflower, sesame, cottonseed and peanut oils, it was superior to the widely utilized analytical chemistry approach in revealing the adulterant species and detecting small quantities of corn and safflower oils in olive oil. Moreover, the DNA-based test correctly identified all tested olive oil: hazelnut oil blends whereas it was not feasible to detect hazelnut oil adulteration through fatty acid profile analysis. Thus, the present research has shown the feasibility of a PCR-CE barcode assay to detect adulteration in olive oil.
European hazelnut (Corylus avellana L.) is an economically and nutritionally important nut crop with wild and cultivated populations found throughout Europe and in parts of Asia. This study examined the molecular genetic diversity and population structure of 402 genotypes including 143 wild individuals, 239 landraces, and 20 cultivars from the Turkish national hazelnut collection using simple sequence repeat (SSR) markers. A total of 30 SSR markers yielded 407 polymorphic fragments. Diversity analysis of the Turkish hazelnut genotypes indicated that they fell into three subpopulations according to ad hoc statistics and neighbor-joining algorithm. Although all cultivars clustered together, they overlapped with the wild accessions and landraces. Thus, the dendrogram, principal coordinate, and population structure analyses suggest that they share the same gene pool. A total of 78 accessions were selected as a core set to encompass the molecular genetic and morphological diversity present in the national collection. This core set should have priority in preservation efforts and in trait characterization.
Tomato leaf miner (Tuta absoluta) is a major pest of tomato in regions with hot climates such as South America and the Mediterranean. This insect feeds on almost every part of the plant and forms galleries while feeding on the plant’s inner tissues. Thus, it can cause plant death and is costly and difficult to control with chemical sprays. In this study, we transferred a modified Bacillus thuringiensis cry1Ac gene to tomato plants via Agrobacterium tumefaciens mediated transformation. Introduction of the cry1Ac gene to the tomato genome was confirmed with PCR and Southern blot analysis in 12 independent events. Insertion sites of the transgene in the tomato genome were determined with TAIL-PCR (thermal asymmetric interlaced polymerase chain reaction) for four selected transgenic lines. Cry1Ac gene expression was verified at both the transcriptional and translational levels, with RT-qPCR and Western blot analyses, respectively. Expression of the Cry1Ac protein in tomato resulted in T. absoluta mortality rates of 38–100% depending on transgenic line. In addition, gallery formation was reduced in 57–100% of the transgenic plants. Moreover, it was found that a single copy of the gene in the hemizygous condition is sufficient to confer tolerance to leaf miner. This is the first reported development of tomato plants resistant to T. absoluta. These transgenic plants are promising for development of commercial tomato cultivars resistant to leaf miner, which will limit the use of environmentally harmful chemicals for control of this pest.
Faba bean (Vivia faba L.) is an important legume species because of its high protein and starch content. Broad bean can be grown in different climatic conditions and is an ideal rotation crop because of the nitrogen fixing bacteria in its roots. In this work, 255 faba bean germplasm accessions were characterized using 32 SSR primers which yielded 302 polymorphic fragments. According to the results, faba bean individuals were divided into two main groups based on the neighbor-joining algorithm (r = 0.91) with some clustering based on geographical origin as well as seed size. Population structure was also determined and agreed with the dendrogram analysis in splitting the accessions into two subpopulations. Analysis of molecular variance (AMOVA) revealed high levels of within population genetic variation. Genetic similarity and geographical proximity were related with separation of European accessions from African and Asian ones. Interestingly, there was no significant difference between landrace (38%) and cultivar (40%) diversity indicating that genetic variability has not yet been lost due to breeding. A total of 44 genetically well-characterized faba bean individuals were selected for a core collection to be further examined for yield and nutritional traits.
Spinach is a popular leafy green vegetable due to its nutritional composition. It contains high concentrations of vitamins A, E, C, and K, and folic acid. Development of genetic markers for spinach is important for diversity and breeding studies. In this work, Next Generation Sequencing (NGS) technology was used to develop genomic simple sequence repeat (SSR) markers. After cleaning and contig assembly, the sequence encompassed 2.5% of the 980 Mb spinach genome. The contigs were mined for SSRs. A total of 3852 SSRs were detected. Of these, 100 primer pairs were tested and 85% were found to yield clear, reproducible amplicons. These 85 markers were then applied to 48 spinach accessions from worldwide origins, resulting in 389 alleles with 89% polymorphism. The average gene diversity (GD) value of the markers (based on a GD calculation that ranges from 0 to 0.5) was 0.25. Our results demonstrated that the newly developed SSR markers are suitable for assessing genetic diversity and population structure of spinach germplasm. The markers also revealed clustering of the accessions based on geographical origin with clear separation of Far Eastern accessions which had the overall highest genetic diversity when compared with accessions from Persia, Turkey, Europe, and the USA. Thus, the SSR markers have good potential to provide valuable information for spinach breeding and germplasm management. Also they will be helpful for genome mapping and core collection establishment.
European hazelnut (Corylus avellana L.), cultivated in several areas of the world including Europe, Anatolia, and the USA, is an economically important nut crop due to its high mineral, oleic acid, amino acid, and phenolic compound content and pleasant flavor. This study examined molecular genetic diversity and population structure of 54 wild accessions and 48 cultivars from the Slovenian national hazelnut collection using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Eleven AFLP primer combinations and 49 SSR markers yielded 532 and 504 polymorphic fragments, respectively. As expected for a wind-pollinated, self-incompatible species, levels of genetic diversity were high with cultivars and wild accessions having mean dissimilarity values of 0.50 and 0.60, respectively. In general, cultivars and wild accessions clustered separately in dendrogram, principal coordinate, and population structure analyses with regional clustering of the wild material. The accessions were also characterized for ten nut and seven kernel traits and some wild accessions were shown to have breeding potential. Morphological principal component analysis showed distinct clustering of cultivars and wild accessions. An association mapping panel composed of 64 hazelnut cultivars and wild accessions had considerable variation for the nut and kernel quality traits. Morphological and molecular data were associated to identify markers controlling the traits. In all, 49 SSR markers were significantly associated with nut and kernel traits [P < 0.0001 and LD value (r2) = 0.15–0.50]. This work is the first use of association mapping in hazelnut and has identified molecular markers associated with important quality parameters in this important nut crop.
Background: Solanum pimpinellifolium has high breeding potential for fruit quality traits and has been used as a donor in tomato breeding programs. Unlocking the genetic potential of S. pimpinellifolium requires high-throughput polymorphism identification protocols for QTL mapping and introgression of favourable alleles into cultivated tomato by both positive and background selection. Results: In this study we identified SNP loci using a genotyping by sequencing (GBS) approach in an IBL mapping population derived from the cross between a high yielding fresh market tomato and S. pimpinellifolium (LA1589) as the recurrent and donor parents, respectively. A total of 120,983,088 reads were generated by the Illumina HiSeq next-generation sequencing platform. From these reads 448,539 sequence tags were generated. A majority of the sequence tags (84.4%) were uniquely aligned to the tomato genome. A total of 3.125 unique SNP loci were identified as a result of tag alignment to the genome assembly and were used in QTL analysis of 11 fruit quality traits. As a result, 37 QTLs were identified. S. pimpinellifolium contributed favourable alleles for 16 QTLs (43.2%), thus confirming the high breeding potential of this wild species. Conclusions: The present work introduced a set of SNPs at sufficiently high density for QTL mapping in populations derived from S. pimpinellifolium (LA1589). Moreover, this study demonstrated the high efficiency of the GBS approach for SNP identification, genotyping and QTL mapping in an interspecific tomato population
Sesame (Sesamum indicum L. syn. Sesamum orientale L.) is considered to be the first oil seed crop known to man. Despite its versatile use as an oil seed and a leafy vegetable, sesame is a neglected crop and has not been a subject of molecular genetic research until the last decade. There is thus limited knowledge regarding genome-specific molecular markers that are indispensible for germplasm enhancement, gene identification, and marker-assisted breeding in sesame. In this study, we employed a genotyping by sequencing (GBS) approach to a sesame recombinant inbred line (RIL) population for high-throughput single nucleotide polymorphism (SNP) identification and genotyping. A total of 15,521 SNPs were identified with 14,786 SNPs (95.26 %) located along sesame genome assembly pseudomolecules. By incorporating sesame-specific simple sequence repeat (SSR) markers developed in our previous work, 230.73 megabases (99 %) of sequence from the genome assembly were saturated with markers. This large number of markers will be available for sesame geneticists as a resource for candidate polymorphisms located along the physical chromosomes of sesame. Defining SNP loci in genome assembly sequences provides the flexibility to utilize any genotyping strategy to survey any sesame population. SNPs selected through a high stringency filtering protocol (770 SNPs) for improved map accuracy were used in conjunction with SSR markers (50 SSRs) in linkage analysis, resulting in 13 linkage groups that encompass a total genetic distance of 914 cM with 432 markers (420 SNPs, 12 SSRs). The genetic linkage map constitutes the basis for future work that will involve quantitative trait locus (QTL) analyses of metabolic and agronomic traits in the segregating RIL population.
The Solanaceae was among the first plant families to be analyzed via comparative mapping and thus was a pioneer in the realm of synteny studies. Analyses of chromosome content and organization have employed a range of techniques, including linkage mapping of genes and molecular markers, physical mapping via fluorescence in situ hybridization, and sequencing of relatively small genomic segments as well as the complete sequencing of the tomato genome. Early comparisons in the family involved tomato and its close relative potato and have extended outward to include eggplant, pepper, tobacco, and petunia. Not surprisingly, the degree of synteny among these species is a function of the time since their divergence, with inversion, translocation, and transposition being the chief mechanisms of chromosome rearrangement. The results of this work provide important insight into the modes and tempo of plant genome evolution while serving a practical purpose as well: knowledge of genome synteny and colinearity makes it easier to leverage resources from one species to another in this agronomically important family.
Natural root-knot nematode resistance genes are unique resources to control this major pest in pepper (Capsicum annuum). Although four genes (Me1, Me3, Me7 and N) conferring broad-spectrum resistance were mapped to a cluster in a 28-cm interval on chromosome P9, limited markers targeting this region were available. In the present study, the Me-gene cluster was structurally annotated for resistance genes to develop markers targeting the N gene. As a result, the Me-gene cluster (4.07 Mb in size) was found to contain three resistance gene hotspots. In addition, a SSR maker tightly linked to the N gene (0.8 cM away) was developed for marker-assisted selection in pepper.
Gene diversity (GD), also called polymorphism information content, is a commonly used measure of molecular marker polymorphism. Calculation of GD for dominant markers such as AFLP, RAPD, and multilocus SSRs is valuable for researchers. To meet this need, we developed a free online computer program, GDdom, which provides easy, quick, and accurate calculation of dominant marker GD with a commonly used formula. Results are presented in tabular form for quick interpretation.
Natural root-knot nematode resistance genes are unique resources to control this major pest in pepper (Capsicum annuum). Although four genes (Me1, Me3, Me7 and N) conferring broad-spectrum resistance were mapped to a cluster in a 28-cm interval on chromosome P9, limited markers targeting this region were available. In the present study, the Me-gene cluster was structurally annotated for resistance genes to develop markers targeting the N gene. As a result, the Me-gene cluster (4.07 Mb in size) was found to contain three resistance gene hotspots. In addition, a SSR maker tightly linked to the N gene (0.8 cM away) was developed for marker-assisted selection in pepper.
Recently developed plant genomics approaches (LD mapping and genome-wide selection) require many molecular markers distributed throughout the plant genome. As a result, the availability of an increasing number of markers is essential for maintaining highly efficient and accurate plant breeding programs. In this study, we identified SNP loci in sunflower using a genotyping by sequencing (GBS) approach in an intraspecific F2 mapping population. A total of 271,445,770 reads were generated by the Genome Analyzer II next-generation sequencing platform and 29.2 % of the reads were aligned to unique locations in the genome. A total of 46,278 SNP loci were identified and 7646 SNP loci were validated in an F2 population. In addition, a SNP-based linkage map was constructed. This is the first report of SNP discovery in sunflower by GBS. The SNP markers and SNP-based linkage map will be valuable molecular genetics tools for sunflower breeding.
Faba bean (Vicia faba L.) is an important food legume crop with a huge genome. Development of genetic markers for faba bean is important to study diversity and for molecular breeding. In this study, we used Next Generation Sequencing (NGS) technology for the development of genomic simple sequence repeat (SSR) markers. A total of 14,027,500 sequence reads were obtained comprising 4,208 Mb. From these reads, 56,063 contigs were assembled (16,367 Mb) and 2138 SSRs were identified. Mono and dinucleotides were the most abundant, accounting for 57.5 % and 20.9 % of all SSR repeats, respectively. A total of 430 primer pairs were designed from contigs larger than 350 nucleotides and 50 primers pairs were tested for validation of SSR locus amplification. Nearly all (96 %) of the markers were found to produce clear amplicons and to be reproducible. Thirty-nine SSR markers were then applied to 46 faba bean accessions from worldwide origins, resulting in 161 alleles with 87.5 % polymorphism, and an average of 4.1 alleles per marker. Gene diversity (GD) of the markers ranged from 0 to 0.48 with an average of 0.27. Testing of the markers showed that they were useful in determining genetic relationships and population structure in faba bean accessions.
Faba bean (Vicia faba L.) is an important food and feed legume because of the nutritional value of its seed protein and starch content, good biomass, and high efficiency nitrogen fixation.This study analyzed the molecular genetic diversity and population structure of 101 Turkish faba bean accessions using simple sequence repeat (SSR) markers.A total of 32 SSR markers yielded 281 loci of which 277 (98.6%) were polymorphic.Mean dissimilarity between genotypes was 0.355, a relatively high value which was expected given faba bean’s large genome and limited breeding history.According to the results, faba bean individuals grouped into three main clusters based on both distance matrix (neighbor-joining algorithm) and model-based (population structure) clustering analyses.Clustering was not correlated with seed size or origin within Turkey.Six cultivars were included in the analysis and showed high genetic diversity compared with the landraces, as expected, given the fact that at least some of the cultivars were bred by hybridization.A total of 47 individuals were selected for the core collection to represent the diversity of Turkish faba bean germplasm.This core set encompasses material adapted to all growing regions and should be a priority for morphological characterization. © 2016, American Society for Horticultural Science.All rights reserved.
As the sole plant source of many potent alkaloids, opium poppy (Papaver somniferum L.) is an important medicinal crop. Nevertheless, few studies have characterized opium poppy germplasm with crop-specific molecular markers. Because Turkey is a diversity center for opium poppy, Turkish germplasm is a valuable genetic resource for association mapping studies aimed at identifying QTLs controlling morphine content and agronomic traits. In this study, the morphological diversity and molecular diversity of 103 Turkish opium poppy landraces and 15 cultivars were analyzed. Potentially useful morphological variation was observed for morphine content, plant height, and capsule index. However, the landraces exhibited limited breeding potential for stigma number, and seed and straw yields. Both morphological and molecular analyses showed distinct clustering of cultivars and landraces. In addition, a total of 164 SSR and 367 AFLP polymorphic loci were applied to an opium poppy association mapping panel composed of 95 opium poppy landraces which were grown for two seasons. One SSR and three AFLP loci were found to be significantly associated with morphine content (P < 0.01 and LD value (r 2) = 0.10–0.32), and six SSR and 14 AFLP loci were significantly associated with five agronomic traits (plant height, stigma number, capsule index, and seed and straw yields) (P < 0.01 and LD value (r 2) = 0.08–0.35). This is the first report of association mapping in this crop. The identified markers provide initial information for marker-assisted selection of important traits in opium poppy breeding.
Glycoalkaloids are plant secondary metabolites that can be both harmful and beneficial to human health. They cause gastroenteric symptoms, coma and even death at high concentrations. It is thought that glycoalkaloids are toxic to human health as a result of their effects on the nervous system and destruction of cell membranes. On the other hand, glycoalkaloids can be effective drugs. For example, solasodine is used to treat skin cancer and tomatidine is used in cancer chemotherapy. Solasodine is also used as a precursor of steroidal drugs. The goal of the work was to isolate and separate efficiently these similar compounds. Materials and methods. Glycoalkaloids from Solanum linnaeanum berries were isolated using column chromatography and confirmed via NMR spectroscopy and MS/MS spectrometry. Results and discussion. The chemical structures of glycoalkaloids are nearly identical. For example, solamargine differs from solasonine in having a methyl group instead of a hydroxyl group bound to the sugar residue of the compound. Thus, their molecular weight and polarity are quite similar. In this work, the process yielded 37.1 mg solasonine, 92.3 mg solasonine and solamargine mix and 56.2 mg solamargine from 81.67 g dried S. linnaeanum berries. Conclusion. Solanum linnaeanum berries are a good source of these glycoalkaloids and the developed protocol proved efficient for purification of solasonine and solamargine.
Reconstructing a tree of life by inferring evolutionary history is an important focus of evolutionary biology. Phylogenetic reconstructions also provide useful information for a range of scientific disciplines such as botany, zoology, phylogeography, archaeology and biological anthropology. Until the development of protein and DNA sequencing techniques in the 1960s and 1970s, phylogenetic reconstructions were based on fossil records and comparative morphological/physiological analyses. Since then, progress in molecular phylogenetics has compensated for some of the shortcomings of phenotype-based comparisons. Comparisons at the molecular level increase the accuracy of phylogenetic inference because there is no environmental influence on DNA/peptide sequences and evaluation of sequence similarity is not subjective. While the number of morphological/physiological characters that are sufficiently conserved for phylogenetic inference is limited, molecular data provide a large number of datapoints and enable comparisons from diverse taxa. Over the last 20 years, developments in molecular phylogenetics have greatly contributed to our understanding of plant evolutionary relationships. Regions in the plant nuclear and organellar genomes that are optimal for phylogenetic inference have been determined and recent advances in DNA sequencing techniques have enabled comparisons at the whole genome level. Sequences from the nuclear and organellar genomes of thousands of plant species are readily available in public databases, enabling researchers without access to molecular biology tools to investigate phylogenetic relationships by sequence comparisons using the appropriate nucleotide substitution models and tree building algorithms. In the present review, the statistical models and algorithms used to reconstruct phylogenetic trees are introduced and advances in the exploration and utilization of plant genomes for molecular phylogenetic analyses are discussed.
Sesame (Sesamum indicum L. syn. Sesamum orientale L.) is an orphan crop species with most molecular genetic research work done in the last decade. In this study, we used a pyrosequenc-ing approach for the development of genomic simple-sequence repeat (SSR) markers in sesame. Our approach proved successful in identifying 19,816 nonredundant SSRs, 5727 of which were identified in a contig assembly that covers 19.29% of the sesame genome. Mononucleotide repeats were the most abundant SSR type identified in the sesame genome (48.5% of all SSRs), followed by dinucleotide SSRs (45.0%). Adenine–thymine-rich motifs were predominant, representing 81.7, 51.7, 66.5, and 22.1% of the mononucleotide, dinucleotide, trinucleotide, and tetranucleo-tide SSRs, respectively. As a result of this work, we introduce 933 experimentally validated sesame specific markers, 849 of which are also applicable in Sesamum mulayanum (syn. Sesamum ori-entale var. malabaricum Nar.), the wild progenitor of cultivated sesame. Using a subset of the newly identified SSR markers, we analyzed molecular genetic diversity and population structure of a collection of world accessions. Results of the two analyses almost overlapped and suggested correlation between genetic similarity and geographical proximity. Indeed, a pattern of gene flow among sesame diversity centers was apparent, with levels of variability in some regions similar to that seen in the domes-tication origin of the crop. Taken together with the high rate of genomic marker transferability detected between S. indicum and S. mulayanum, our results represent additional molecular genetic evidence for designating the two taxa as cultivated and wild forms of the same species.
Root knot nematodes (Meloidogyne spp.) are significant agricultural pests on many crops, including pepper (Capsicum annuum). Host plant resistance offers the most sustainable means of controlling this pest. A cluster of genes on chromosome 9 confers resistance, with Me1 providing protection against three nematode species: M. incognita, M. javanica and M. arenaria. We describe the development of a codominant CAPS marker located 1.13 cM away from the Me1 gene. This marker should be useful for marker assisted selection of nematode resistance in pepper breeding programs.
The aim of this study was to develop a DNA-barcode assay to authenticate the botanical origin of herbal teas. To reach this aim, we tested the efficiency of a PCR-Capillary Electrophoresis (PCR-CE) approach on commercial herbal tea samples using two non-coding plastid barcodes, the trnL intron and the intergenic spacer between trnL and trnF. Barcode DNA length polymorphisms proved successful in authenticating the species origin of herbal teas. We verified the validity of our approach by sequencing species-specific barcode amplicons from herbal tea samples. Moreover, we displayed the utility of PCR-CE assays coupled with sequencing to identify the origin of undeclared plant material in herbal tea samples. The PCR-CE assays proposed in this work can be applied as routine tests for the verification of botanical origin in herbal teas and can be extended to authenticate all types of herbal foodstuffs.
The conservation of plant germplasm is essential to ensure future breeding gains and crop bio-diversity. To be truly useful, such germplasm must be characterized for morphological traits and genetic diversity. In this work, agro-morphological diversity was assessed in 137 Turkish sesame (Sesamum indicum L.) genotypes (129 accessions and eight cultivars) by examination of eight qualitative and nine quantitative traits. As expected, morphological variability in the cultivars was low with broader diversity present in sesame accessions. However, some accessions were identified with interesting features, such as increased number of capsules and seed yield, which could be employed in future cultivar development. The sesame genotypes were analyzed for molecular genetic diversity with 140 amplified fragment length polymorphism (AFLP) loci. The results indicated a relatively low level of variability with an average dissimilarity value of 0.33 for all genotypes. Population structure was also examined and indicated that the material fell into two subpopulations. As most of the accessions (82%) were obtained from the U.S. Department of Agriculture (USDA) and are not yet housed in the Turkish national sesame germplasm collection, the data were used to identify a core set of 22 accessions that should be preserved in Turkey. The importance of using both molecular and morphological data for core selection is highlighted with a focus on germplasm preservation and breeding.
The aim of this study was to establish a DNA-based identification key to ascertain the cultivar origin of Turkish monovarietal olive oils. To reach this aim, we sequenced short fragments from five olive genes for SNP (Single Nucleotide Polymorphism) identification and developed CAPS (Cleaved Amplified Polymorphic DNA) assays for SNPs that alter restriction enzyme recognition motifs. When applied on the oils of 17 olive cultivars, a maximum of five CAPS assays were necessary to discriminate the varietal origin of the samples. We also tested the efficiency and limit of our approach for detecting olive oil admixtures. As a result of the analysis, we were able to detect admixing down to a limit of 20%. The SNP-based CAPS assays developed in this work can be used for testing and verification of the authenticity of Turkish monovarietal olive oils, for olive tree certification, and in germplasm characterization and preservation studies.
Wild tomato species have been widely used for improvement of tomato disease resistance but have not been extensively explored for health-related traits. In this work, three interspecific populations derived from backcrosses between cultivated tomato and Solanum pimpinellifolium (LA1589), S. habrochaites (LA1223), and S. peruvianum (LA2172) were analyzed for water-soluble antioxidant activity, phenolic content, vitamin C content, and basic agronomic traits including fruit weight, shape, and color. The wild species accessions significantly exceeded S. lycopersicum for all three antioxidant traits with only one exception: vitamin C content in S. habrochaites LA1223. Several populations and traits showed transgressive segregation indicating that the backcross populations contained individuals with allele combinations that allowed antioxidant activity/content to exceed that of both parents. The S. habrochaites LA1223 population provided the best starting material for improvement of water-soluble antioxidant activity and phenolics content with 20% and 15% of the population, respectively, significantly exceeding the parental values for these traits. Moreover, the S. habrochaites population contained individuals that had nearly 2-fold more water-soluble antioxidant activity and phenolic content than cultivated tomato. The S. peruvianum LA2172 population was best for improvement of vitamin C content with 3-fold variation for the trait and individuals, which had twice as much vitamin C as cultivated tomato.
Fifty-eight F2 individuals derived from an interspecific cross between cultivated eggplant, Solanum melongena, and its wild relative, S. linnaeanum, were phenotyped for 42 plant, leaf, flower, and fruit traits. Composite interval mapping analysis using genotypic data from 736 molecular markers revealed the positions of 71 statistically significant (P ≤ 0.05) quantitative trait loci (QTL) influencing 32 of the morphological traits. Although most QTL were location-specific, QTL governing three traits (leaf lobing, leaf prickles and prickle anthocyanin) were detected in both experimental locations. Analysis of three additional traits (stem prickles, fruit calyx prickles and fruit length) in both locations yielded QTL in similar but non-overlapping map positions. The majority (69 %) of the QTL corresponded closely with those detected in previous analyses of this data set. However the increased resolution of the linkage map combined with advances in QTL mapping permitted more precise localization, such that the average interval length of these QTL was reduced by 93 %. Thirty-one percent of the QTL were novel, suggesting that simple linear regression with a low density linkage map (the method used in previous studies of this population) missed a substantial portion of significant QTL. Hotspots of QTL affecting plant hairiness, prickliness, and pigmentation were identified on chromosomes 3, 6, and 10, respectively, and may reflect the pleiotropic activity of single structural or regulatory genes at these positions. Based on synteny between the eggplant, tomato, potato and pepper genomes, putative orthologs were identified for 35 % of the QTL suggesting strong conservation of gene function within the Solanaceae. These results should make it easier to target particular loci for map-based cloning and marker-assisted selection studies.
A linkage map of eggplant was constructed for an interspecific F 2 population derived from a cross between Solanum linnaeanum MM195 and S. melon-gena MM738. The map contains 400 AFLP Ò (ampli-fied fragment length polymorphism), 348 RFLP (restriction fragment length polymorphism) and 116 COSII (conserved ortholog set) markers. The 864 mapped markers encompass 12 linkage groups, span 1,518 cM and are spaced at an average interval of 1.8 cM. Use of orthologous markers allowed confir-mation of the established syntenic relationships between eggplant and tomato chromosomes and helped delineate the nature of the 33 chromosomal rearrangements and 11 transpositions distinguishing the two species. This genetic map provides a 2-to 3-fold improvement in marker density compared to previously published interspecific maps. Because the interspecific mapping population is rich in morpho-logical variation, this greater genome saturation will be useful for QTL (quantitative trait locus) analyses. The recent release of the tomato genome sequence will provide additional opportunities for exploiting this map for comparative genomics and crop improvement.
Opium poppy (Papaver somniferum L.) is an important pharmaceutical crop with very few genetic marker resources. To expand these resources, we sequenced genomic DNA using pyrosequencing technology and examined the DNA sequences for simple sequence repeats (SSRs). A total of 1,244,412 sequence reads were obtained covering 474 Mb. Approximately half of the reads (52 %) were assembled into 166,724 contigs representing 105 Mb of the opium poppy genome. A total of 23,283 non-redundant SSRs were identified in 18,944 contigs (11.3 % of total contigs). Trinucleotide and tetranucleotide repeats were the most abundant SSR repeats, accounting for 49.0 and 27.9 % of all SSRs, respectively. The AAG/TTC repeat was the most abundant trinucleotide repeat, representing 19.7 % of trinucleotide repeats. Other SSR repeat types were AT-rich. A total of 23,126 primer pairs (98.7 % of total SSRs) were designed to amplify SSRs. Fifty-three genomic SSR markers were tested in 37 opium poppy accessions and seven Papaver species for determination of polymorphism and transferability. Intraspecific polymorphism information content (PIC) values of the genomic SSR markers were intermediate, with an average 0.17, while the interspecific average PIC value was slightly higher, 0.19. All markers showed at least 88 % transferability among related species. This study increases sequence coverage of the opium poppy genome by sevenfold and the number of opium poppy-specific SSR markers by sixfold. This is the first report of the development of genomic SSR markers in opium poppy, and the genomic SSR markers developed in this study will be useful in diversity, identification, mapping and breeding studies in opium poppy.
With almost 870 million people estimated to suffer fromchronic hunger worldwide, undernourishment represents amajor problem that severely affects people in developing countries. In addition to undernourishment, micronutrient deficiency alone can be a cause of serious illness and death. Large portions of the world population rely on a single, starch-rich crop as their primary energy source and these staple crops are generally not rich sources of micronutrients. As a result, physical and mental health problems related to micronutrient deficiencies are estimated to affect around two billion people worldwide. The situation is expected to get worse in parallelwith the expandingworld population. Improving the nutritional quality of staple crops seems to be an effective and straightforward solution to the problem. Conventional breeding has long been employed for this purpose but success has been limited to the existing diversity in the gene pool. However, biotechnology enables addition or improvement of any nutrient, even those that are scarce or totally absent in a crop species. In addition, biotechnology introduces speed to the biofortification process compared to conventional breeding. Genetic engineering was successfully employed to improve a wide variety of nutritional traits over the last decade. In the present review, progress toward engineering various types of major and minor constituents for the improvement of plant nutritional quality is discussed.
All publicly available opium poppy expressed sequence tag (EST) sequences, totalling 20 885, were assembled into unigenes and examined for simple sequence repeats (SSRs). Nearly 19% of the 14 957 unigenes contained SSRs with 4% harbouring more than one SSR. Average den-sity of the SSRs was 1 SSR per 3.6 kb of non-redundant EST sequence. Trinucleotide SSRs were most frequently identified (39%), and many of the most prevalent motifs were AT-rich. Flanking primers were designed for 86% of the SSRs and 67 primer pairs were tested on 37 opium poppy accessions and seven related species. All markers were transferable to the related species. Polymorphism information content (PIC) values for the markers were intermediate for comparisons within opium poppy (average of 0.27) and slightly higher for comparisons across species (average of 0.29). The markers were found to be useful for diversity analysis as they successfully distinguished among Turkish opium poppy accessions and land races.
Turkey is a secondary center of diversity for melon ( Cucumis melo ) and is home to a variety of regional morphotypes. This diversity is housed in a national germplasm repository with more than 500 accessions. Molecular genetic variability of 209 melon genotypes from 115 accessions of this collection was characterized using amplified fragment length polymorphisms (AFLPs). Ten AFLP primer combinations yielded 279 reproducible fragments, which were used for dendrogram and principal coordinate analyses. These analyses showed two major clusters of Turkish melons: one group contained highly similar genotypes (maximum Dice dissimilarity coefficient of 0.18), whereas the other group was genetically more diverse (maximum dissimilarity 0.41). Although average dissimilarity was low (0.13), a broad range of genetic diversity was observed in the collection. A marker allele richness strategy was used to select a core set of 20 genotypes representing the allelic diversity of the AFLP data. The core set had double the average diversity (0.26) of the entire set and represented the major morphotypes present in the collection. Molecular genetic diversity of the core set was further validated using simple sequence repeat marker data (116 polymorphic fragments), which confirmed that the selected core set retained high levels of molecular genetic diversity.
Resistance of eggplant against Ralstonia solanacearum phylotype I strains was assessed in a F(6) population of recombinant inbred lines (RILs) derived from a intra-specific cross between S. melongena MM738 (susceptible) and AG91-25 (resistant). Resistance traits were determined as disease score, percentage of wilted plants, and stem-based bacterial colonization index, as assessed in greenhouse experiments conducted in Réunion Island, France. The AG91-25 resistance was highly efficient toward strains CMR134, PSS366 and GMI1000, but only partial toward the highly virulent strain PSS4. The partial resistance found against PSS4 was overcome under high inoculation pressure, with heritability estimates from 0.28 to 0.53, depending on the traits and season. A genetic map was built with 119 AFLP, SSR and SRAP markers positioned on 18 linkage groups (LG), for a total length of 884 cM, and used for quantitative trait loci (QTL) analysis. A major dominant gene, named ERs1, controlled the resistance to strains CMR134, PSS366, and GMI1000. Against strain PSS4, this gene was not detected, but a significant QTL involved in delay of disease progress was detected on another LG. The possible use of the major resistance gene ERs1 in marker-assisted selection and the prospects offered for academic studies of a possible gene for gene system controlling resistance to bacterial wilt in solanaceous plants are discussed.
The antioxidant potential of 42 melon (Cucumis melo) lines including six cultivars grown in Turkey was assessed by measuring total water-soluble antioxidant capacity, phenolic and vitamin C contents. The lines showed significant variability for all three antioxidant parameters with breeding lines having higher antioxidant capacity and phenolic content than some popular cultivars. Different types of melons also showed significantly different antioxidant potentials. Thus, galia and ananas types showed a higher mean antioxidant capacity and phenolic content than the other tested types (yuva, kislik, canary, and charentais). Correlation analysis between antioxidant parameters showed a significant correlation between water-soluble antioxidant capacity and phenolic content.
The genetic diversity and relatedness of 90 watermelon (Citrullus lanatus var. lanthus and Citrullus. lanatus var. citroides) accessions were molecularly characterized using 30 sequence-related amplified polymorphism (SRAP) marker combinations. The accessions were collected from Turkey (59 accessions) and other regions of the world by the United States Department of Agriculture (USDA) and the molecular data were analysed using UPGMA (Unweighted Pair Group Method Analysis). The SRAP combinations were highly polymorphic (97%) with 87 polymorphic bands determined among 201 amplified fragments (43%). The UPGMA analysis characterized five major clusters (A, B, C, D and E). While minimum genetic similarity among groups A, B and C was high (83%), group E was the most distant with 63% genetic similarity. Principal component analysis (PCA) was performed and used to produce a two-dimensional plot from which two main groups could be distinguished. Based on the analyses, genetic diversity of watermelons was very low and Turkish watermelons were not distinct from other countries' accessions. These results could be used for generation of a core collection of watermelon by elimination of redundant accessions and for watermelon breeding by helping to identify useful, genetically distinct lines. In addition, the study indicated that SRAP markers are useful for analysing genetic diversity in crops like watermelon which have low variability.
The growth and ion content of salt sensitive Lycopersicon esculentum Mill. cv. M82 and salt tolerant L. pennellii Correll accession LA716 were examined under both control and stress conditions (150 mM NaCl). L. esculentum grew more vigorously than L. pennellii under optimal conditions, however, L. pennellii was able to maintain its growth better than cultivated tomato when the plants were exposed to salinity. Sodium content of both L. esculentum and L. pennellii increased as a result of NaCl stress. In addition, both species showed reduced potassium and calcium content due to salinity. The physiological traits were also measured in a population of 52 L. pennellii introgression lines grown under both normal and stress conditions. A total of 311 quantitative trait loci (QTL) were identified for the studied traits: plant height, stem diameter, leaf number, leaf and root fresh and dry mass, and sodium, potassium and calcium contents. Some of the loci (124) were identified under both control and stress conditions while 86 QTL were identified only under non-stress conditions and 101 loci were identified only under NaCl stress. Additional key wordscalcium– Lycopersicon esculentum –potassium–salinity–sodium–tomato
In Turkey, local varieties of eggplant (Solanum melongena L.) occur in several types (including round, semi-long and long) and are a staple ingredient of the cuisine. Although Turkish eggplant varieties are morphologically distinct, little is known about their molecular genetic variation. In this study, the genetic variability of 67 Turkish eggplant accessions from the national germplasm collection was assessed with 30 morphological traits and AFLP markers. Morphological analysis indicated considerable variability especially for semi-long and round types. For molecular characterization, accessions of S. macrocarpon, S. aethiopicum and S. linnaeanum were included as outgroups. Ten primer combinations were used and yielded 488 polymorphic fragments with PIC values ranging from 0.03 to 0.50. Of the polymorphic fragments, 144 (29%) were specific to S. melongena accessions while 73, 49 and 16 fragments were specific to S. macrocarpon, S. aethiopicum and S. linnaeanum, respectively. UPGMA cluster analysis of the AFLP data resulted in a dendrogram which had a very high correlation (r=0.97) with the similarity matrix data. Genetic similarity in the dendrogram ranged from 0.30 to 0.95 with the related Solanum species located outside the S. melongena clusters, as expected. Genetic similarity of the S. melongena accessions ranged from 0.68 to 0.95 indicating good genetic diversity present in the Turkish national collection. It is hoped that this information, together with morphological data will help guide future germplasm collection and eggplant breeding efforts.
Resistance of eggplant against Ralstonia solanacearum phylotype I strains was assessed in a F 6 population of recombinant inbred lines (RILs) derived from a intra-specific cross between S. melongena MM738 (susceptible) and AG91-25 (resistant). Resistance traits were determined as disease score, percentage of wilted plants, and stem-based bacterial colonization index, as assessed in greenhouse experiments conducted in Réunion Island, France. The AG91-25 resistance was highly efficient toward strains CMR134, PSS366 and GMI1000, but only partial toward the highly virulent strain PSS4. The partial resistance found against PSS4 was overcome under high inoculation pressure, with heritability estimates from 0.28 to 0.53, depending on the traits and season. A genetic map was built with 119 AFLP, SSR and SRAP markers positioned on 18 linkage groups (LG), for a total length of 884 cM, and used for quantitative trait loci (QTL) analysis.
Within the genus Solanum, the term 'eggplant' encompasses several cultivated species that are used for food and, to a lesser extent, for medicine. The use of one common name to describe more than one species and the existence of many related wild species have led to taxonomic confusion which, in turn, have complicated analyses of evolutionary relationships and genetic diversity within this groups of species. A further challenge for eggplant research is that, despite the fact that the use of molecular markers for phylogenetic studies is well-established, very few studies have described the development of new markers for eggplant. In our work, genic microsatellite (SSR) marker; were identified from an expressed sequence tag library of S. melongena and used for analysis of 47 accessions of eggplant and closely related species. The markers had very good polymorphism in the 18 species tested including 8 S. melongena accessions. Moreover, genetic analysis performed with these markers showed concordance with previous research and knowledge of eggplant domestication. These markers are expected to be a valuable resource for studies of genetic relationships, fingerprinting, and gene mapping in eggplant.
Tomato is one of the most widely produced and consumed vegetable crops worldwide. Plant breeders have usually focused on improvement of horticulturally important traits such as yield, fruit size, shape and colour. With increased attention on human health, however, plant breeders also consider the improvement of health-related traits of fruits and vegetables such as antioxidant characters. In the present study, genes controlling both health-related and horticulturally important traits were mapped in the tomato genome using 152 Lycopersicon hirsutum BC2F2 individuals. For this aim, all plants were phenotypically and genotypically characterised and a total of 75 QTLs were identified for all traits. Of the 75 QTLs, 28 were identified for 5 antioxidant traits including total water soluble antioxidant capacity, vitamin C, total phenolics, total flavonoids, and lycopene contents, and 47 QTLs were identified for 8 agronomic traits including fruit weight, external and internal fruit colour, fruit firmness, fruit shape, stem scar size, locule number, and wall thickness. Markers linked with these QTLs can be used in marker assisted selection (MAS) for improvement of elite tomato lines.
Bacterial wilt, caused by strains belonging to the Ralstonia solanacearum species complex, inflicts severe economic losses in many crops worldwide. Host resistance remains the most effective control strategy against this disease. However, wilt resistance is often overcome due to the considerable variation among pathogen strains. To help breeders circumvent this problem, we assembled a worldwide collection of 30 accessions of tomato, eggplant and pepper (Core-TEP), most of which are commonly used as sources of resistance to R. solanacearum or for mapping quantitative trait loci. The Core-TEP lines were challenged with a core collection of 12 pathogen strains (Core-Rs2) representing the phylogenetic diversity of R. solanacearum. We observed six interaction phenotypes, from highly susceptible to highly resistant. Intermediate phenotypes resulted from the plants' ability to tolerate latent infections (i.e., bacterial colonization of vascular elements with limited or no wilting). The Core-Rs2 strains partitioned into three pathotypes on pepper accessions, five on tomato, and six on eggplant. A "pathoprofile" concept was developed to characterize the strain clusters, which displayed six virulence patterns on the whole set of Core-TEP host accessions. Neither pathotypes nor pathoprofiles were phylotype specific. Pathoprofiles with high aggressiveness were mainly found in strains from phylotypes I, IIB, and III. One pathoprofile included a strain that overcame almost all resistance sources.
Olive (Olea europaea L.) is one of the most important and characteristic fruit crops of the Mediterranean region. This crop has been grown in Turkey for more than 3000 yr and many varieties are currently grown in five geographic regions. In this study, the genetic diversity of 66 olive varieties from these regions was assessed using simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) markers. The 13 SSR markers yielded 89 alleles with an average polymorphism information content (PIC) of 0.29 and 6.8 alleles per marker while the 13 SRAP primer combinations generated 103 polymorphic alleles with an average PIC of 0.24 and 7.9 alleles per combination. Although SSR markers revealed higher levels of polymorphism than SRAP markers, both systems revealed considerable molecular genetic diversity in Turkish olive varieties. For both marker types, clustering analysis using the Dice similarity coefficient and the unweighted pair group method with arithmetic means (UPGMA) produced dendrograms with similar clustering and some region-specific grouping of varieties. Overall, Southeast Anatolian and Marmara varieties were found to be more genetically different than those from the other regions (Aegean, Mediterranean, and Black Sea) and homonymous varieties were identified: three pairs of varieties ('Egriburun', 'Celebi', and 'Tasarasi') had the same names but were genetically distinct.
Growth characteristics and antioxidant content of M82, LA716 and IL lines under control conditions and salt stress. For M82 and LA716, salt effect refers to the fold change in trait/activity observed when lines were subject to salt stress as compared to control conditions. Salt effect values are only included for those differences which were statistically significant as determined by Student's t-test (P < 0.05). Nonsignificant effects are indicated by "ns", "na' indicates that statistical analysis was not appropriate because replicates were bulked. For the ILs, salt effect is the percentage of ILs showing significant increases and decreases in each parameter under salt stress as compared to nonstress conditions.
Loci identified for the antioxidant traits in the IL population. Control and salt-specific QTL names are suffixed with "c" and "s" respectively. Effect refers to the phenotypic effect (percent change relative to M82) of the S. pennellii allele for each locus.
Excessive soil salinity is an important problem for agriculture, however, salt tolerance is a complex trait that is not easily bred into plants. Exposure of cultivated tomato to salt stress has been reported to result in increased antioxidant content and activity. Salt tolerance of the related wild species, Solanum pennellii, has also been associated with similar changes in antioxidants. In this work, S. lycopersicum M82, S. pennellii LA716 and a S. pennellii introgression line (IL) population were evaluated for growth and their levels of antioxidant activity (total water-soluble antioxidant activity), major antioxidant compounds (phenolic and flavonoid contents) and antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase and peroxidase) under both control and salt stress (150 mM NaCl) conditions. These data were then used to identify quantitative trait loci (QTL) responsible for controlling the antioxidant parameters under both stress and nonstress conditions. Under control conditions, cultivated tomato had higher levels of all antioxidants (except superoxide dismutase) than S. pennellii. However, under salt stress, the wild species showed greater induction of all antioxidants except peroxidase. The ILs showed diverse responses to salinity and proved very useful for the identification of QTL. Thus, 125 loci for antioxidant content under control and salt conditions were detected. Eleven of the total antioxidant activity and phenolic content QTL matched loci identified in an independent study using the same population, thereby reinforcing the validity of the loci. In addition, the growth responses of the ILs were evaluated to identify lines with favorable growth and antioxidant profiles. Plants have a complex antioxidant response when placed under salt stress. Some loci control antioxidant content under all conditions while others are responsible for antioxidant content only under saline or nonsaline conditions. The localization of QTL for these traits and the identification of lines with specific antioxidant and growth responses may be useful for breeding potentially salt tolerant tomato cultivars having higher antioxidant levels under nonstress and salt stress conditions.
Species of the Ceranisus thrips-attacking genus are difficult to distinguish morphologically. The phylogenetic relationships within the Ceranisus species were explored using nucleotide sequences of the 28S D2 expansion region of the rDNA gene. Bayesian, maximum likelihood, and parsimony inference methods were employed to construct the phylogenetic relationships. Principal component analysis on the Turkish species of Ceranisus, namely antalyacus, menes, bozovaensis, hirsutus, planitianus (a new record for Turkey), pacuvius, and a new species, provided supporting evidence. All known data concerning hosts and geographical distribution are presented. A new species, C. onuri O. Doganlar, sp.n., was described from Turkey. Türkiye thrips parazitoitlerinin bazı morfolojik ölçümler ve 28S D2 rDNA özelliklerine dayanarak oluşturulan ön filogenisi ile yeni bir türün tanısı Özet: Thripsler üzerinde doğal düşman olan, Ceranisus cinsinin morfolojik olarak teşhis edilmeleri oldukça zordur. Bu sebeple çalışmada bu böceklerin rDNA'sına ait genişlemiş 28S D2 bölgesinin gen dizisi kullanılarak, Ceranisus türleri için bir filogenetik ilişki ortaya konmuştur. Filogenetik ilişki oluşturulurken Bayesian, Maximum likelihood ve Parsimony metodları kullanılmıştır. Morfolojik ölçümler kullanılarak Ceranisus, viz., antalyacus, menes, bozovaensis, hirsutus, planitianus (Türkiye için yeni kayıt), pacuvius ve yeni bir tür üzerinde temel bileşenler analizi yapılmış ve sonuçlar bu yeni türün farklı tür olduğunu desteklemiştir. Türler üzerinde coğrafik yayılış, ilişkide olduğu konak bitkiler ile ilgili bilinen tüm bilgiler verilmiştir. Türkiye' den yeni bir tür C. onuri O. Doganlar, sp.n. tanımlanmıştır.
- Jan 2010
- 9èmes rencontres Plantes-Bactéries, 18-22 janvier 2010 : Aussois 2010, du 18 au 22 janvier 2010. - Paris : SFP, 20109èmes rencontres Plantes-Bactéries, 18-22 janvier 2010 : Aussois 2010, du 18 au 22 janvier 2010. - Paris : SFP, 2010
Ralstonia solanacearum est responsable de phytobactérioses vasculaires très destructrices, aussi bien en zone tropicale que tempérée. Parmi les rares stratégies de lutte disponibles, la plus efficace est la résistance variétale. Cette dernière est souvent partielle chez les Solanées à graines (tomate, aubergine, piment), s'exprime de façon quantitative et peut être contournée par certaines souches de la bactérie. En effet, la résistance variétale interagit avec la forte diversité génétique et phénotypique reconnue dans le complexe d'espèce R. solanacearum (4 phylotypes). La conséquence est que la résistance fluctue d'une zone géographique à l'autre. Nous avons choisi 12 souches représentatives de la diversité de R. solanacearum (collection Core-Rs2), et les avons inoculées à 30 accessions couvrant les résistances variétales les plus pertinentes connues chez ces 3 espèces (collection Core-TEP). En chambre climatique, ces interactions Core-Rs2 x Core-TEP se déclinent en 6 phénotypes, allant de très sensible à très résistant. La résistance chez certains cultivars d'aubergine peut être totale. Chez le piment, les infections latentes (colonisation sans flétrissement) sont majoritaires, ce qui suggère des mécanismes de défense différents de ceux présents chez la tomate et l'aubergine. L'analyse de ces interactions permettent de définir 6 groupes de souches, appelés `pathoprofils', dont l'un est constitué d'une souche qui contourne pratiquement toutes les résistances connues. Aucun pathoprofil n'apparaît phylotype-spécifique. Les deux pathoprofils les plus virulents sont composés en majorité de souches de phylotypes IIB (Amériques) et III (Afrique). Cette étude ambitieuse est la première à confronter la variabilité connue de R. solanacearum à la diversité des ressources génétiques pour la résistance chez ces trois espèces. (Résumé d'auteur)
The aim of this study was to investigate genetic analysis of the species within genus Ceranisus (Hymenoptera: Eulophidae) that are interesting groups found in South Eastern Anatolia and Eastern parts of Mediterranean Region of Turkey and to compare with the other thrips-parasitoids between May 2006 and May 2007. Totally, 143 specimens (30 male: 113 female) belonging to Ceranisus genus were collected from 20 locations of 38 different localities in Hatay, Gaziantep, Adiyaman and Şanliurfa provinces. The population was divided into 13 groups based on morphologic characters. Phylogenetic relationships amongst Ceranisus species (Chalcidoidea: Eulophidae) were gathered from DNA sequence data. Fifteen species of thrips chalcid parasitoids were analyzed using one gene, D2 domain of the 28S ribosomal DNA. Among morphologically distinct 13 species/taxonomic groups, 3 of them identified as new species according to the genetical analysis. Four of the other species occurred in C. nr. lepidotus group while 5 of C. nr. bozovaensis consisted of two groups. To conclude 3 new species of Ceranisus and 3 different groups that may yield new species were determined in Turkish Ceranisus population.
In this study, total water soluble antioxidant activity and phenolic content of 26 eggplant (Solanum melongena L.) cultivars were investigated. Total water soluble antioxidant activity of the cultivars varied from 2664 to 8247 μmolTrolox/kg, which is a 3.1-fold difference. Cultivars also showed significant variation for total phenolic contents ranging from 615 to 1376 mg/kg, a 2.2-fold difference. The two traits were significantly correlated and results of this study suggested that breeders can use the information to develop eggplant cultivars with high antioxidant activity.
Potato Y potyvirus (PVY) is an important plant pathogen worldwide that infects and causes yield losses in the family Solanaceae including potato (Solarium tuberosum), pepper (Capsicum spp.), tomato (S. lycopersicum), and tobacco (Nicotiana tabacum). In this study, 20 different tomato accessions representing 6 different species were mechanically inoculated with PVY°. The plants were scored visually for symptoms and then tested for presence of the virus 2-4 weeks after inoculation by ELISA. The results were variable. Most wild species of tomato sustained PVY° replication in inoculated leaves. Some of the wild species showed an immune response, while some became systemically infected. Inoculation and analysis of F2 populations suggested that the resistance is controlled by a single recessive gene in different wild species.
In this work, 29 pepper cultivars that represent the diversity of types and varieties grown in Turkey were analyzed for water-soluble antioxidant capacity and phenolic and vitamin C contents. In addition, 14 non-Turkish cultivars were tested for comparison. Significant diversity was observed in the different cultivars with the most variation (7.4-fold) seen for total antioxidant capacity, which ranged from 2.57 to 18.96 mmol Trolox/kg. Vitamin C content for the peppers ranged from 522 to 1631 mg.kg(-1), a 3.1 -fold difference, whereas total phenolic content for the pepper cultivars ranged from 607 to 2724 mg-kg(-1), a 4.5-fold difference. When cultivars were grouped by morphology/use, it was found that some types had significantly more variation and higher antioxidant activities than other types. Thus, for water-soluble antioxidant capacity, most variation was seen in long, blunt-ended Carliston types, whereas long, pointed Sivri peppers had the highest mean capacity. Bell-shaped Dolmalik and Sivri peppers had the most variation for phenolic content, but fancy Sus and Sivri types had the highest means for this trait. Dolmalik types showed the most variation for vitamin C content, whereas Sus and Sivri peppers had the highest means for this character. All three parameters were significantly and positively correlated with the strongest correlation between total antioxidant capacity and phenolic content (r = 0.71). The presence of significant variation for antioxidant content in Turkish germplasm indicates that this material can be used for improvement and genetic mapping of nutritional content in pepper.
A practical set of HPLC methods was developed for the separation and determination of the eggplant steroidal glycoalkaloids, solanine, chaconine, solasonine, solamargine, and their aglycones, solasodine and solanidine. A gradient method was initially developed, but proved to be neither robust nor practical. Three separate isocratic methods using acetonitrile and ammonium dihydrogen phosphate were developed and shown to be more repeatable, less subject to fluctuations in mobile phase composition, and less time consuming. The effect of adjusting buffer pH, column temperature, and buffer type (triethylammonium phosphate vs. ammonium dihydrogen phosphate) were evaluated. It was also discovered that, by addition of 10% methanol to the acetonitrile portion of the mobile phase, more control over the separations was possible. The use of methanol as a mobile phase entrainer greatly improved separations in some cases and its effectiveness was also dependent upon column temperature. Assessments of the method recovery, limit of detection, and limit of quantitation were made using extracts from S. melongena and S. linnaeanum.
Tomato and potato expressed sequence tag (EST) sequences contained in the solanaceae genomics network (SGN) database were screened for simple sequence repeat (SSR) motifs. A total of 609 SSRs were identified and assayed on Solanum lycopersicum LA925 (formerly Lycopersicon esculentum) and S. pennellii LA716 (formerly L. pennellii). The SSRs that did not amplify, gave multiple band products, or did not exhibit a polymorphism that could be readily detected on standard agarose gels in either of these species were eliminated. A set of 76 SSRs meeting these criteria was then placed on the S. lycopersicum (LA925) x S. pennellii (LA716) high-density map. A set of 76 selected cleaved amplified polymorphism (CAP) markers was also developed and mapped onto the same population. These 152 PCR-based anchor markers are uniformly distributed and encompass 95% of the genome with an average spacing of 10.0 cM. These PCR-based markers were further used to characterize S. pennellii introgression lines (Eshed and Zamir, Genetics 141:1147-1162, 1995) and should prove helpful in utilizing these stocks for high-resolution mapping experiments. The majority of these anchor markers also exhibit polymorphism between S. lycopersicum and two wild species commonly used as parents for mapping experiments, S. pimpinellifolium (formerly L. pimpinellifolium) and S. habrochaites (formerly L. hirsutum), indicating that they will be useful for mapping in other interspecific populations. Sixty of the mapped SSRs plus another 49 microsatellites were tested for polymorphism in seven tomato cultivars, four S. lycopersicum var. cerasiforme accessions and eight accessions of five different wild tomato species. Polymorphism information content values were highest among the wild accessions, with as many as 13 alleles detected per locus over all accessions. Most of the SSRs (90%) had accession-specific alleles, with the most unique alleles and heterozygotes usually found in accessions of self-incompatible species. The markers should be a useful resource for qualitative and quantitative trait mapping, marker-assisted selection, germplasm identification, and genetic diversity studies in tomato. The genetic map and marker information can be found on SGN (http://www.sgn.cornell.edu).
Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10-15%.
Eggplant fruit color is highly variable, both for qualitative as well as quantitative aspects. Pigment chemistry explains part of the many nuances which are discernible by eye between green fruited cultivars, and even more between the so called "violet-purple" fruited cultivars. Phenotypic complexity of eggplant fruit colour (at commercial stage) can be reduced into four main parametres (1) anthocyanin production in fruit epidermis, (2) anthocyanin production under the calyx, (3) chlorophyll production in fruit flesh, and (4) cholorphyll distribution (uniform, reticulated,...). Colour measurement methods are numerous, but the end-user in accordance to the type materials used and to the research purpose. Some crosses made recently at INRA provided suitable segregation progenies for dissecting the inheritance of the main fruit colour components. Anthocyanin presence (vs its absence) is under monogenic dominant control (gene provisionally symbolized A), Anthocyanin presence under the calyx (vs its absence) is under the control of a monogenic dominant gene (symbol Puc). Green flesh is monogenic dominant (symbol G) over the white flesh. The reticulated distribution of chlorophylls (the not reticulated one) is under monogenic dominant control (symbol Gv). Some of these genetic factors have been already located on the eggplant molecular map, and putative orthologous counterparts in other Solanaceae identified.
Despite their unsuitability for agricultural production, the wild relatives of crop species represent a largely untapped resource of novel QTLs potentially useful for crop plant improvement. In this regard, previous introgression studies, involving several different wild tomato species, have shown that the long arm of chromosome 4 contains QTLs for many horticulturally important traits including soluble solids content, fruit shape, lycopene content and biochemical composition. However, these earlier studies were unable to determine how many genes control these traits and whether genes affecting the same character from different wild species are allelic or not. In an effort to shed light on these issues,we have constructed a series of lines containing small, overlapping introgressions for portions of the long arm of chromosome 4 from L. peruvianum and L. hirsutum and tested these lines in replicated field trials. The results provide evidence for multiple, non-allelic loci controlling soluble solids and fruit weight. They also show that the loci controlling some traits (e.g. fruit shape, fruit weight, epidermal reticulation) co-localize to the same portions of chromosome 4, a result that maybe attributed to pleiotropy and/or gene dense areas with lower than average recombination. The implications of these finding for molecular breeding and utilization of exotic germplasm are discussed.
In this study, the advanced backcross QTL (AB-QTL) mapping strategy was used to identify loci for yield, processing and fruit quality traits in a population derived from the interspecific cross Lycopersicon esculentum E6203 x Lycopersicon pennellii accession LA1657. A total of 175 BC(2) plants were genotyped with 150 molecular markers and BC(2)F(1) plots were grown and phenotyped for 25 traits in three locations in Israel and California, U.S.A. A total of 84 different QTLs were identified, 45% of which have been possibly identified in other wild-species-derived populations of tomato. Moreover, three fruit-weight/size and shape QTLs ( fsz2b.1, fw3.1/ fsz3.1 and fs8.1) appear to have putative orthologs in the related solanaceous species, pepper and eggplant. For the 23 traits for which allelic effects could be deemed as favorable or unfavorable, 26% of the identified loci had L. pennellii alleles that enhanced the performance of the elite parent. Alleles that could be targeted for further introgression into cultivated tomato were also identified.
- Jan 2004
- Transgenic Plants
Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10–15%.
An interspecific F(2) population from a cross between cultivated eggplant, Solanum melongena, and its wild relative, S. linnaeanum, was analyzed for quantitative trait loci (QTL) affecting leaf, flower, fruit and plant traits. A total of 58 plants were genotyped for 207 restriction fragment length polymorphism (RFLP) markers and phenotyped for 18 characters. One to eight loci were detected for each trait with a total of 63 QTL identified. Overall, 46% of the QTL had allelic effects that were the reverse of those predicted from the parental phenotypes. Wild alleles that were agronomically superior to the cultivated alleles were identified for 42% of the QTL identified for flowering time, flower and fruit number, fruit set, calyx size and fruit glossiness. Comparison of the map positions of eggplant loci with those for similar traits in tomato, potato and pepper revealed that 12 of the QTL have putative orthologs in at least one of these other species and that putative orthology was most often observed between eggplant and tomato. Traits showing potential orthology were: leaf length, shape and lobing; days to flowering; number of flowers per inflorescence; plant height and apex, leaf and stem hairiness. The functionally conserved loci included a major leaf lobing QTL ( llob6.1) that is putatively orthologous to the potato leaf ( c) and/or Petroselinum ( Pts) mutants of tomato, two flowering time QTL ( dtf1.1, dtf2.1) that also have putative counterparts in tomato and four QTL for trichomes that have potential orthologs in tomato and potato. These results support the mounting evidence of conservation of gene function during the evolution of eggplant and its relatives from their last common ancestor and indicate that this conservation was not limited to domestication traits.
The near-isogenic line (NIL) TA1150 contains a 56-cM introgression from Lycopersicon chmielewskii chromosome 1 and has several interesting phenotypic characteristics including fruit with orange color, high levels of soluble solids, thick pericarp, small stem scars, and good firmness. A set of overlapping recombinant lines (subNILs) was developed and field tested to fine map the quantitative trait loci (QTL) controlling these traits. The results indicated that the solids, pericarp thickness, and firmness QTL are distinct from the color locus. Several of the QTL mapped in this study, including the soluble-solids QTL, probably correspond to QTL mapped in other wild species of tomato. However, analysis of a set of TA523 subNILs containing complementary introgressions from Lycopesicon hirsutum chromosome 1 suggests that this wild species may contain a different locus for improved soluble solids. Thus, it might be possible to combine the L. chmielewskii and L. hirsutum alleles for these loci in a single line with the potential for extremely highly soluble solids. The TA1150 subNIL TA1688 contains the smallest introgression of the solids locus (approximately 19 cM), as well as the pericarp thickness and firmness QTL, with a yield that was equivalent to two of the three control lines. Isolation of recombinant subNILs from TA1688 should break the linkage between orange color and high solids and provide a small introgressed segment for marker-assisted breeding and genetic improvement of processing tomato.
Although tomato has been the subject of extensive quantitative trait loci (QTLs) mapping experiments, most of this work has been conducted on transient populations (e.g., F2 or backcross) and few homozygous, permanent mapping populations are available. To help remedy this situation, we have developed a set of inbred backcross lines (IBLs) from the interspecific cross between Lycopersicon esculentum cv. E6203 and L. pimpinellifolium (LA1589). A total of 170 BC2F1 plants were selfed for five generations to create a set of homozygous BC2F6 lines by single-seed descent. These lines were then genotyped for 127 marker loci covering the entire tomato genome. These IBLs were evaluated for 22 quantitative traits. In all, 71 significant QTLs were identified, 15% (11/71) of which mapped to the same chromosomal positions as QTLs identified in earlier studies using the same cross. For 48% (34/71) of the detected QTLs, the wild allele was associated with improved agronomic performance. A number of new QTLs were identified including several of significant agronomic importance for tomato production: fruit shape, firmness, fruit color, scar size, seed and flower number, leaf curliness, plant growth, fertility, and flowering time. To improve the utility of the IBL population, a subset of 100 lines giving the most uniform genome coverage and map resolution was selected using a randomized greedy algorithm as implemented in the software package MapPop (http://www.bio.unc.edu/faculty/vision/lab/ mappop/). The map, phenotypic data, and seeds for the IBL population are publicly available (http://soldb.cit.cornell.edu) and will provide tomato geneticists and breeders with a genetic resource for mapping, gene discovery, and breeding.
Domesticated species differ from their wild ancestors and relatives for a set of traits that is known as the domestication syndrome. The most important syndrome traits include growth habit, flowering time, seed dispersal, gigantism and morphological diversity. This paper reviews what is known about the genetic control of domestication traits with an emphasis on comparative analyses that examine this control in two or more crop species. Such analyses indicate that although most domestication traits are quantitatively controlled, the dramatic morphological changes that accompanied domestication may be due to relatively few genes. These studies also show that domestication genes have been functionally conserved over thousands of years and have similar, although not identical, effects in various species.
A molecular genetic linkage map based on tomato cDNA, genomic DNA, and EST markers was constructed for eggplant, Solanum melongena. The map consists of 12 linkage groups, spans 1480 cM, and contains 233 markers. Comparison of the eggplant and tomato maps revealed conservation of large tracts of colinear markers, a common feature of genome evolution in the Solanaceae and other plant families. Overall, eggplant and tomato were differentiated by 28 rearrangements, which could be explained by 23 paracentric inversions and five translocations during evolution from the species' last common ancestor. No pericentric inversions were detected. Thus, it appears that paracentric inversion has been the primary mechanism for chromosome evolution in the Solanaceae. Comparison of relative distributions of the types of rearrangements that distinguish pairs of solanaceous species also indicates that the frequency of different chromosomal structural changes was not constant over evolutionary time. On the basis of the number of chromosomal disruptions and an approximate divergence time for Solanum, approximately 0.19 rearrangements per chromosome per million years occurred during the evolution of eggplant and tomato from their last ancestor. This result suggests that genomes in Solanaceae, or at least in Solanum, are evolving at a moderate pace compared to other plant species.
Quantitative trait loci (QTL) for domestication-related traits were identified in an interspecific F(2) population of eggplant (Solanum linnaeanum x S. melongena). Although 62 quantitative trait loci (QTL) were identified in two locations, most of the dramatic phenotypic differences in fruit weight, shape, color, and plant prickliness that distinguish cultivated eggplant from its wild relative could be attributed to six loci with major effects. Comparison of the genomic locations of the eggplant fruit weight, fruit shape, and color QTL with the positions of similar loci in tomato, potato, and pepper revealed that 40% of the different loci have putative orthologous counterparts in at least one of these other crop species. Overall, the results suggest that domestication of the Solanaceae has been driven by mutations in a very limited number of target loci with major phenotypic effects, that selection pressures were exerted on the same loci despite the crops' independent domestications on different continents, and that the morphological diversity of these four crops can be explained by divergent mutations at these loci.
The efficiency of the binary bacterial artificial chromosome (BIBAC) vector for Agrobacterium-mediated stable transfer of high molecular weight DNA into plants was tested in tomato. Several variables affecting transformation efficiency were examined including insert size, Agrobacterium genetic background, and the presence of additional copies of the virG, virE1 and virE2 genes. It was found that a helper plasmid containing extra copies of virG was an absolute requirement for obtaining tomato transformants with the BIBAC. MOG101 with the virG helper plasmid was found to be the most efficient strain for transfer of high molecular weight DNA (150 kb). Selected high molecular weight DNA transformants were advanced several generations (up to the R4) to assess T-DNA stability. This analysis showed that the T-DNA was stably maintained and inherited through several meioses regardless of whether it was in the hemizygous or homozygous state. Expression of a selectable marker gene within the T-DNA was also examined through several generations and no gene silencing was observed. Thus, the BIBAC is a useful system for transfer of large DNA fragments into the plant genome.
- Jan 2001
- DNA-Based Markers in Plants
Cultivated tomato (Lycopersicon esculentum L.) is a member of the nightshade family (Solanaceae) which includes several other crop plants including potato (Solanum tuberosum), pepper (Capsicum annuum), eggplant (S. melongena), and tobacco (Nicotiana tabacum). Of these solanaceous species, tomato is the most amenable to genetic analysis as it is diploid (2n = 2x = 24), autogamous and has a relatively small nuclear genome of approximately 950Mb (Arumuganathan and Earle 1991). The tomato genome contains nearly 80% single copy sequences as determined by high stringency hybridization conditions (Zamir and Tanksley 1988), a characteristic which also facilitates genetic analysis. The remainder of the genome is comprised of repetitive DNA. Repetitive elements that have been characterized and mapped in tomato include ribosomal DNA (Bernatzky and Tanksley 1986; Vallejos et al. 1986; Lapitan et al. 1991), the tomato genomic repeats TGRI, TGRII and TGRIII (Ganal et al. 1988; Lapitan et al. 1989), minisatellites (Broun and Tanksley 1993; Broun and Tanksley 1996), and microsatellites (Broun and Tanksley 1996; Vosman and Arens 1997).
We used a positional cloning approach to isolate the Sw-5 disease resistance locus of tomato. Complementation experiments with overlapping cosmid clones enabled us to demonstrate that Sw-5 is a single gene locus capable of recognizing several tospovirus isolates and species. Analysis of the predicted Sw-5 protein suggests that it is a cytoplasmic protein, with a potential nucleotide binding site (NBS) domain and a C-terminal end consisting of leucine-rich repeats (LRRs). Based on its structural features, Sw-5 belongs to the class of NBS-LRR resistance genes that includes the tomato Mi, 12, and Prf genes; the Arabidopsis RPM1 gene; and the plant potato virus X resistance gene Rx. The overall similarity between the Sw-5 and Mi proteins of tomato suggests that a shared or comparable signal transduction pathway leads to both virus and nematode resistance in tomato. The similarity also supports the hypothesis that Sw-5 provides resistance via a hypersensitive response. Sw-5 is a member of a loosely clustered gene family in the telomeric region of chromosome 9. Members of this family map to other regions of chromosome 9 and also to chromosome 12, where several fungal, virus, and nematode genes have been mapped, suggesting that paralogs of Sw-5 may have evolved to provide different resistance specificities.
Domestication of many plants has correlated with dramatic increases in fruit size. In tomato, one quantitative trait locus (QTL), fw2.2, was responsible for a large step in this process. When transformed into large-fruited cultivars, a cosmid derived from the fw2.2 region of a small-fruited wild species reduced fruit size by the predicted amount and had the gene action expected for fw2.2. The cause of the QTL effect is a single gene, ORFX, that is expressed early in floral development, controls carpel cell number, and has a sequence suggesting structural similarity to the human oncogene c-H-ras p21. Alterations in fruit size, imparted by fw2.2 alleles, are most likely due to changes in regulation rather than in the sequence and structure of the encoded protein.
The seeds of domesticated plants are normally much larger than those of their wild counterparts. This change in seed weight was most likely in response to the selection pressure for yield, uniform germination and seedling vigor which was exerted by humans during domestication. However, despite the evolutionary and agronomic significance of seed weight, very little is know about the genetic and developmental controls of this trait; and, thus far, none of the genes in this pathway have been isolated from any plant species. QTL mapping experiments conducted in tomato during the past decade have allowed the identification of many seed-weight QTLs and have also revealed that only a few loci are responsible for the majority of the seed-weight changes that accompanied the domestication of tomato. This review presents a consensus map for seed weight QTL identified in previously published reports and in unpublished results from our laboratory. This summary of seed-weight QTL data allows for the identification of the major loci controlling this trait in the genus Lycopersicon. It is hoped that this work will allow the elucidation of this important phenotypic transition that occurred during crop-plant domestication and will also provide the starting point for the cloning of a gene responsible for seed-weight variation.
- Apr 1999
This is the first report of large insert genomic DNA libraries constructed in a binary-BAC (BIBAC) vector. Genomic DNA libraries containing approximately 4.6 haploid nuclear genomic equivalents were constructed for Lycopersicon esculentum (cv. Mogeor) and Lycopersicon pennellii (LA716) in the BIBAC2 vector. The L. esculentum library has an average insert size of 125 kb and is comprised of 42 272 individual colonies stored as frozen cultures in a 384-well format (108 plates). The L. pennellii library has an average insert size of 90 kb and is comprised of 53 760 individual clones (140 384-well plates). In each of the libraries, it is estimated that 90% of the colonies contain genomic DNA inserts. The composition of the L. esculentum and L. pennellii libraries was determined by analyzing a series of randomly selected clones. The L. esculentum library was surveyed for clones containing chloroplast DNA (1.4%), mitochondrial DNA (0.012%) and repetitive DNA motifs. BIBAC clones that may contain a gene of interest can be identified from these libraries by colony hybridization with homologous or heterologous probes or by PCR pooling techniques. Once identified, BIBAC genomic DNA library clones are immediately suitable for Agrobacterium tumefaciens-mediated plant transformation.
Two accessions of Lycopersicon peruvianum Mill. (PI270435, PI126443) carrying Mi-2 and Mi-3, respectively, new root-knot nematode resistance genes, were selected as the male parents for crosses with L. esculentum Mill. in order to produce interspecific hybrids. Crossability barriers between these two distantly related species were circumvented by ovule culture. A total of ten interspecific F1 hybrid plants were produced. The hybrid nature of the putative F1 plants was confirmed by a comparison of several morphological characteristics and a PCR-based assay. Eight of ten hybrid plants were backcrossed with L. esculentum to generate a total of 98 BC1 progeny. Two lines were advanced to the BC2 and BC3 levels. Based on these results, ovule culture was found to be an effective method for the production of novel interspecific F1 and BC1 plants.
An improved protocol forAgrobacterium-mediated transformation of the tomato cultivar Moneymaker was developed by examining the effects of six different factors on the efficiency of transformation. Explant size, explant orientation, gelling agent and plate sealant were found to affect transformation efficiency. Two other factors, type of explant (hypocotyl or cotyledon) and frequency of transfer to fresh selective regeneration medium, did not have any effect on transformation efficiency. By combining the best treatments for each factor, an average transformation efficiency of 10.6% was obtained for Moneymaker.
In conjunction with an enhanced system for Agrobacterium-mediated plant transformation, a new binary bacterial artificial chromosome (BIBAC) vector has been developed that is capable of transferring at least 150 kb of foreign DNA into a plant nuclear genome. The transferred DNA appears to be intact in the majority of transformed tobacco plants analyzed and is faithfully inherited in the progeny. The ability to introduce high molecular weight DNA into plant chromosomes should accelerate gene identification and genetic engineering of plants and may lead to new approaches in studies of genome organization.
- Jan 1995
Tomato (Lycopersicon esculentum) cultivars that conatin the Pto bacterial resistance locus also exhibit sensitivity to fenthion, an organophosphorous insecticide. Resistance to Pseudomonas syringae pv. tomato (Pst) encoded by the Pto gene and sensitivity to fenthion cosegregate in large F2 populations and were apparently introgressed together into tomato from the wild species Lycopersion pimpinellifolium. In order to isolate the genes responsible for these two phenotypes and to study their molecular basis, a multistep map-based cloning project was initiated. A closely linked RFLP marker was used to isolate a yeast artificial chromosome (YAC) clone that spanned the Pto region. Transcribed sequences within the Pto region were identified by isolating cDNA clones that hybridized to the YAC clone. Transformation of candidate cDNA clones into a Pst-susceptible, fenthion-insensitive tomato line succeeded in identifying a cDNA conferring Pst resistance and a separate cDNA conferring sensitivity to fenthion. The cDNA clones represent members of a clustered gene family and encode putative serine-threonine protein kinases. The role of these kinases in recognizing a signal from Pst or the fenthion molecule and in activating the plant response is currently being investigated.
Leaves of tomato cultivars that contain the Pto bacterial resistance locus develop small necrotic lesions within 24 hr after exposure to fenthion, an organophosphorous insecticide. Recently, the Pto gene was isolated and shown to be a putative serine/threonine protein kinase. Pto is one member of a multigene family that is clustered within a 400-kb region on chromosome 5. Here, we report that another member of this gene family, termed Fen, is responsible for the sensitivity to fenthion. Fen was isolated by map-based cloning using closely linked DNA markers to identify a yeast artificial chromosome clone that spanned the Pto region. After transformation with the Fen gene under control of the cauliflower mosaic virus (CaMV) 35S promoter, tomato plants that are normally insensitive to fenthion rapidly developed extensive necrotic lesions upon exposure to fenthion. Two related insecticides, fensulfothion and fenitrothion, also elicited necrotic lesions specifically on Fen-transformed plants. Transgenic tomato plants harboring integrated copies of the Pto gene under control of the CaMV 35S promoter displayed sensitivity to fenthion but to a lesser extent than did wild-type fenthion-sensitive plants. The Fen protein shares 80% identity (87% similarity) with Pto but does not confer resistance to Pseudomonas syringae pv tomato. These results suggest that Pto and Fen participate in the same signal transduction pathway.
The Pto gene in tomato confers resistance to races of Pseudomonas syringae pv. tomato that carry the avirulence gene avrPto. A yeast artificial chromosome clone that spans the Pto region was identified and used to probe a leaf complementary DNA (cDNA) library. A cDNA clone was isolated that represents a gene family, at least six members of which genetically cosegregate with Pto. When susceptible tomato plants were transformed with a cDNA from this family, they were resistant to the pathogen. Analysis of the amino acid sequence revealed similarity to serine-threonine protein kinases, suggesting a role for Pto in a signal transduction pathway.
Thesis (Ph. D.)--Cornell University, August, 1995. Includes bibliographical references.
Typescript. Honors paper - Mount Holyoke College, 1990. Department of Biological Sciences. Includes bibliographical references (leaves 66-69).