Using randomly amplified polymorphic DNA for evaluating genetic relationships among papaya cultivars

Summary We have applied the recently developed technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among ten cultivars of papaya (Carica papaya L.). Eleven ten-base synthetic oligonucleotides were chosen that gave multiple PCR amplification products using papaya DNA as template. These 11 primers amplified a total of 102 distinct fragments. Cultivars were scored for presence or absence of RAPD fragments and grouped by cluster analysis using simple matching coefficients of similarity. A dendrogram of the ten cultivars was constructed. Of the ten cultivars seven were of the Hawaiian type, and all of these grouped to one branch of the tree. Divisions within the Hawaiian, branch were mostly consistent with the known genetic background of these cultivars. Three non-Hawaiian, cultivars were also analyzed. The minimum similarity detected was 0.7 suggesting that the domesticated papaya germ plasm is quite narrow. Our results show that RAPD technology is a rapid, precise and sensitive technique for genomic analysis.Journal series No. 3692 of the Hawaii Institute of Tropical Agriculture and Human Resources     

六个品种花椰菜花球的营养成分分析与评价

为评价花椰菜(Brassica oleracea var. botrytis)的营养品质,测定了6个品种花球的10项营养指标,运用主成分分析和聚类分析方法对花椰菜品质进行综合评价。结果表明,6个品种的10项品质指标均存在不同程度的差异,变异幅度为12.22%~ 131.21%。维生素C (Vc)、总黄酮、总多酚、Fe、Ca、P、蛋白质含量间存在显著或极显著的关联性。系统聚类分析将6种花椰菜分为4类,黄色花椰菜‘209’、‘100’和‘217’各为1类,白色花椰菜‘210’、‘214’和‘218’聚为1类。主成分分析提取了花椰菜品质综合评价的3个主成分,获得6个营养评价指标：Vc、总黄酮、总多酚、Fe、Ca和P。通过建立评价函数模型: F= 0.5591Z₁+0.2189Z₂+0.1669Z₃,筛选出‘209’花椰菜的营养品质最高。这为挖掘及选育优良花椰菜品种提供了科学依据。     

Molecular phylogeny and DNA amplification fingerprinting of Petunia taxa

The relationship of five species of Petunia and ten cultivars of the cultivated petunia, Petunia x hybrida, were investigated using DNA-amplification fingerprinting (DAF). Reproducible banding profiles were obtained from P. parodii and P. axillaris DNA from different seed sources. In contrast, other petunias such as P. inflata, P. violacea and P. integrifolia produced variable fingerprints when different plants were examined. However, representative profiles of the variable Petunia taxa were obtained by bulking the leaf tissue from ten different individual plants. Each of ten octamer primers revealed polymorphic loci between taxa. Among a total of 201 bands produced, 146 (73%) loci were polymorphic and distinguished all species and cultivars. Phenetic and cluster analysis using DAF markers separated P. axillaris from P. parodii and distinguished between the violet-flowered species, P. inflata, P. violacea, and P. integrifolia. P. parodii grouped together with the monophyletic set of the ten cultivars of P. x hybrida examined, indicating that it had made a major contribution to the development of these cultivars. Cultivars were distributed within the dendograms by flower color. The results demonstrated the utility of DAF in establishing relationships among closely related species and cultivars of Petunia.     

Genetic diversity and population structure among cultivars of Saccharina japonica currently farmed in northern China

Close to 50% of the annual production of the brown alga Saccharina japonica, about 2 million tons (fresh weight), is produced through farming in Shandong province, China, principally around Rongcheng. Under artificial selection for phenotypic traits (e.g. color, blade length) cultivated strains (cultivars) face the problems of inbreeding depression, which can be caused by the close relationships of the parental plants. Hence, in an attempt to evaluate genetic variations and relationships among cultivars, 15 major farmed cultivars were selected and sampled from seven major hatcheries and analyzed using 15 microsatellite markers. A total of 94 alleles were found across all samples, with allele numbers ranging from three to 17 per locus. All of the cultivars exhibited relatively high levels of genetic diversity, with mean Nei's genetic diversity (H) and the Shannon's information index (I) of 0.466 and 0.862, respectively. A UPGMA dendrogram grouped all cultivars into three main clusters. However, two of the most commonly farmed cultivars (DB and BN) from different hatcheries failed to cluster together. Instead, differently named cultivars from the same hatchery tended to group together, implying that their high similarities is partly due to the presence of genetic mixing among cultivars within a hatchery or misuse of cultivar's names. Genetic analysis performed by Bayesian model‐based clustering revealed clear differentiation of three major subgroups (LJ‐202 and LJ‐205; LJ‐C033; XS‐2 and XS‐BN) and one admixed group (the remaining ten cultivars). Our results revealed ambiguous genetic relationships among certain cultivars of S. japonica farmed in northern China. A more stringent and prudent regulation should be applied during breeding and production process in the future.     

Resistance to Asparagus virus 1 in the Wild Relative Asparagus amarus

Five asparagus cultivars, three breeding lines and the wild relative Asparagus amarus were tested for natural infection by Asparagus virus 1 (AV‐1) in experimental fields at two locations over 3 and 4 years, respectively. In the first year after re‐planting the annual crowns in the field, more than 90% of tested plants of cultivars were infected by AV‐1. In the third and fourth year, 100% of tested plants of cultivars were AV‐1 infected. In comparison, all plants of the wild relative A. amarus were completely free of AV‐1, suggesting a high level of resistance. Additionally, 1‐year‐old glasshouse‐cultivated plants of A. officinalis and A. amarus were placed in an AV‐1 provocation cabin under field conditions. Seven months later, 100% of the A. officinalis plants showed a high virus concentration in ELISA, whereas no AV‐1 was detectable in the A. amarus plants. This result was confirmed by highly sensitive AV‐1‐specific RT‐PCR. To exclude vector resistance, the feeding behaviour of green peach aphid Myzus persicae was tested over 12 h using the electrical penetration graph method. Both asparagus genotypes were accepted by the aphids as potential hosts, but the feeding time was significantly longer on A. amarus. A genetic distance analysis of the various cultivars of Asparagus officinalis and selected wild relatives of the JKI collection was carried out, resulting in a clear discrimination of cultivars and wild relatives, especially A. amarus. The potential breeding value of the putative resistance carrier is discussed.     

基于Pacbio第三代测序技术的厚朴基因组测序分析

厚朴为著名的传统药用植物,归于木兰科、木兰属,于我国广泛种植,其树皮、根皮、枝皮、叶片、花、果实均能入药或食用。为获取厚朴全基因组序列信息,该文以厚朴叶片DNA为材料,采用Pacbio Sequel第三代测序技术构建厚朴全基因组数据库,并利用生物信息学方法对获得的核苷酸序列进行组装、功能注释以及进化分析研究。结果表明:(1)原始测序数据过滤后获得140.91 Gb三代数据,Read N50约为13 784bp,经过组装得到厚朴基因组大小为1.68 Gb,Contig N50约为222 069 bp,单拷贝基因完整性为81.0%。(2)组装后的序列通过与NR、KOG、KEGG等功能数据库比对,共有98.40%的基因得到了功能注释,其中KOG功能注释结果发现厚朴的蛋白功能主要集中在一般功能预测、翻译后修饰、蛋白质转换、伴侣以及信号转导机制; GO功能分类表明厚朴的基因集中在细胞组分及生物学过程; KEGG分析发现厚朴参与代谢通路的基因占主要地位。(3)通过与葡萄、拟南芥、水稻、杨树、银杏、无油樟、茶树及牛樟基因组的比对分析,发现厚朴23 424个基因中有20 801个基因可以分类到12 129个家族,其中有515个基因家族为厚朴所特有,而厚朴与牛樟(樟科)亲缘关系较近,两者的分化时间约在122.5百万年前(mya)。该研究首次利用第三代测序技术对厚朴全基因组解析,有利于对其进一步进行深入的开发与利用,也为研究其他药用植物全基因组奠定了基础。     

Total phenolic content and antioxidant activity of plants used in traditional Romanian herbal medicine

A number of herbal plants from Romania widely used as natural food additives or for health promotion in traditional medicine were investigated for their antioxidant activity. Methanol extracts were obtained from plants belonging to the Lamiaceae family (lavender Lavandula angustifolia L.; lemon balm Melissa officinalis; sage Salvia officinalis; oregano Origanum vulgare L.; rosemary Rosmarinus officinalis L.; thyme Thymus vulgaris L.; mullein Verbascum phlomoides; mint Mentha longifolia), Clusiaceae family (St John’s wort Hypericum perforatum L.), and Compositae family (elecampane Inula helenium). Total phenolic concentration was determined using the Folin-Ciocalteu phenol reagent method, while total flavonoids were measured using the aluminium chloride colorimetric method. Relationships between total antioxidant activity and composition of plant extracts were evaluated. Origanum vulgare extract showed the highest antioxidant activity and total phenolic content compared to the other plants extracts. A positive correlation was observed between total antioxidant activity and total phenolic content of the analyzed extracts.     

Molecular phylogeny of the genus Asparagus (Asparagaceae) explains interspecific crossability between the garden asparagus (A. officinalis) and other Asparagus species

The genus Asparagus comprises approximately 200 species, some of which are commercially cultivated, such as the garden asparagus (A. officinalis). Many Asparagus species, including A. officinalis, are dioecious and have been grouped into a subgenus distinct from that of hermaphroditic species. Although many interspecific crossings have been attempted to introduce useful traits into A. officinalis, only some of the dioecious species were found to be cross-compatible with A. officinalis. Here, molecular phylogenetic analyses were conducted to determine whether interspecific crossability is proportional to the genetic distance between the crossing pairs and to further clarify the evolutionary history of the Asparagus genus. A clade with all cross-compatible species and no cross-incompatible species was recovered in the phylogenetic tree based on analyses of non-coding cpDNA regions. In addition, a sex-linked marker developed for A. officinalis amplified a male-specific region in all cross-compatible species. The phylogenetic analyses also provided some insights about the evolutionary history of Asparagus; for example, by indicating that the genus had its origin in southern Africa, subsequently spreading throughout the old world through intensive speciation and dispersal. The results also suggest that dioecious species were derived from a single evolutionary transition from hermaphroditism in Asparagus. These findings not only contribute towards the understanding of the evolutionary history of the genus but may also facilitate future interspecific hybridization programs involving Asparagus species.     

Microbiological and toxicological quality of dried herbs

Aims: The microbiological and toxicological quality of 51 samples of dried herbs (Melissa officinalis, Salvia officinalis, Malva sylvestris, Matricaria chamomilla, Alchemilla vulgaris and Centaurea cyanus) cultivated in family‐managed farms in Molise Region (Italy) was evaluated. Methods and Results: All the samples were analysed by using conventional methods, and for samples preparation, an alternative Washing and Shaking (WaS) protocol was developed to reduce release of antimicrobial compounds. None of the samples were of unsatisfactory quality with respect to aflatoxin B1, and only three samples from Malva sylvestris exceeded the limit of total aflatoxins according to Recommendation 2004/24/EC. The International Commission on Microbiological Specifications for Foods limits for mesophilic bacteria and total coliforms were exceeded in the 29·4 and 3·9% of samples, respectively: 7·8% of samples also exceeded the limit for Escherichia coli established by European Spice Association. When the ‘WaS’ method was used, higher microbial counts were obtained, especially for A. vulgaris, S. officinalis and M. officinalis. Conclusions: Herbs cultivated in family‐managed small agricultural areas showed a good microbiological and toxicological quality, irrespectively of preliminary washing or selection procedures. Significance and Impact of the Study: Herb matrices may contain antimicrobial activity which should be considered when applying the conventional microbiological methods for sample preparation. Alternative preparation protocols may have advantages to reduce antimicrobial effects and should be further evaluated.     

Identification of simple sequence repeats (SSRs) in olive ( Olea europaea L.)

A small insert genomic library of Olea europaea L., highly enriched in (GA/CT)n repeats, was obtained using the procedure of Kandpal et al. (1994). The sequencing of 103 clones randomly extracted from this library allowed the identification of 56 unique genomic inserts containing simple sequence repeat regions made by at least three single repeats. A sample of 20 primer pairs out of the 42 available were tested for functionality using the six olive varieties whose DNA served for library construction. All primer pairs succeeded in amplifying at least one product from the six DNA samples, and ten pairs detecting more than one allele were used for the genetic characterisation of a panel of 20 olive accessions belonging to 16 distinct varieties. A total of 57 alleles were detected among the 20 genotypes at the ten polymorphic SSR loci. The remaining primer pair allowed the amplification of a single SSR allele for all accessions plus a longer fragment for some genotypes. Considering the simple sequence repeat polymorphism, 5.7 alleles were scored on average for each of the ten SSR loci. A genetic dissimilarity matrix, based on the proportion of shared alleles among all the pair-wise combinations of genotypes, was constructed and used to disentangle the genetic relationships among varieties by means of the UPGMA clustering algorithm. Graphical representation of the results showed the presence of two distinct clusters of varieties. The first cluster grouped the varieties cultivated on the Ionian Sea coasts. The second cluster showed two subdivisions: the first sub-cluster agglomerated the varieties from some inland areas of Calabria; the second grouped the remaining varieties from Basilicata and Apulia cultivated in nearby areas. Results of cluster analysis showed a significant relationship between the multilocus genetic similarities and the geographic origin of the cultivars. Received: 2 February 2001 / Accepted: 1 June 2001     

Analysis of genetic variability in two diploid <Emphasis Type="Italic">Musa</Emphasis> cultivars using RAPD

Twenty accessions of the sparsely cultivated diploid Musa cultivars Matti (AA) and Rasakadali (AB) were subjected to random amplified polymorphic DNA (RAPD) assay. A total of 14 random primers were used for the estimation of interand intracultivar variations. Out of 86 bands generated, 64 were polymorphic (74.4 % polymorphism). The cluster analysis grouped the cultivars into two major clusters: cluster I with 10 accessions of Matti and 2 of Rasakadali and cluster II comprising the remaining 8 accessions of Rasakadali. The coefficient of genetic similarity (GS) was from 0.73 to 0.99, suggesting low level of intercultivar variation. The accessions of Rasakadali with mean GS of 0.89 were genetically more diverse than those of Matti (GS = 0.93).     

Essential Oils and Chemical Diversity of Southeast European Populations of Salvia officinalis L.

The essential oils of 25 populations of Dalmatian sage (Salvia officinalis L.) from nine Balkan countries, including 17 indigenous populations (representing almost the entire native distribution area) and eight non‐indigenous (cultivated or naturalized) populations were analyzed. Their essential‐oil yield ranged from 0.25 to 3.48%. Within the total of 80 detected compounds, ten (β‐pinene, 1,8‐cineole, cis‐thujone, trans‐thujone, camphor, borneol, trans‐caryophyllene, α‐humulene, viridiflorol, and manool) represented 42.60 to 85.70% of the components in the analyzed essential oils. Strong positive correlations were observed between the contents of trans‐caryophyllene and α‐humulene, α‐humulene and viridiflorol, and viridiflorol and manool. Principal component analysis (PCA) on the basis of the contents of the ten main compounds showed that four principal components had an eigenvalue greater than 1 and explained 79.87% of the total variation. Performing cluster analysis (CA), the sage populations could be grouped into four distinct chemotypes (A–D). The essential oils of 14 out of the 25 populations of Dalmatian sage belonged to Chemotype A and were rich in cis‐thujone and camphor, with low contents of trans‐thujone. The correlation between the essential‐oil composition and geographic variables of the indigenous populations was not significant; hence, the similarities in the essential‐oil profile among populations could not be explained by the physical proximity of the populations. Additionally, the southeastern populations tended to have higher EO yields than the northwestern ones.     

Microsatellites and microsynteny in the chloroplast genomes of Oryza and eight other Gramineae species

Primer pairs flanking ten chloroplast microsatellite loci, originally identified in Oryza sativa cv Nipponbare, were evaluated for amplification and allelic diversity using a panel of 13 diverse cultivars of rice (O. sativa), 19 accessions of wild rice (three O. officinalis, five O. latifolia, five O. minuta, four O. australiensis, one O. brachyantha and one O. ridleyi) and eight other Gramineae species (maize, teosinte, wheat, oat, barley, pearl millet, sorghum and sugarcane). Amplified products were obtained for all samples at nine out of ten loci. Among the rice cultivars, the number of alleles per locus ranged from one to four, with monomorphic patterns observed at five loci. The average polymorphism information content (PIC) value at the other five (polymorphic) loci was 0.54 among the 13 cultivars. When wild rice and the other Gramineae species were compared based on the proportion of shared alleles, their phylogenetic relationships were in agreement with previous studies using different types of markers; however, the magnitude of the differences based on chloroplast microsatellites underestimated the genetic distance separating these divergent species and genera. A sequence-based comparison of homologous regions of the rice and maize chloroplast genomes revealed that, while a high level of microsynteny is evident, the occurrence of actively evolving microsatellite motifs in specific regions of the rice chloroplast genome appears to be mainly a species or genome-specific phenomenon. Thus the chloroplast primer pairs used in this study bracketed mutationally active microsatellite motifs in rice but degenerate, interrupted motifs or highly conserved, mutationally inert motifs in distantly related genera. Received: 17 March 1999 / Accepted: 11 November 1999     

Discrimination of Polygonati Rhizoma Species: An Investigation Utilizing High-Performance Liquid Chromatography Fingerprints and Chemometrics

Polygonati Rhizoma has been a famous traditional Chinese medicine (TCM) for two thousand years. It is increasingly being used not just as a traditional herbal medicine but also as a popular functional food. In this study, qualitative and quantitative analysis of PR from three different origins were initially performed using chemical fingerprint and chemometrics methods. Hierarchical cluster analysis (HCA) and Principal component analysis (PCA) were used to classify 60 PR samples from three different origins. The results revealed that the PR samples fell into three clusters related to the origins. In addition, pairwise comparison of varying PR and obtaining chemical markers between different species through the establishment of partial least squares discriminant analysis. Finally, chemical markers 9,13 and 17 were identified by LC/MS as disporopsin, 5,7-dihydroxy-3-(4′-hydroxybenzyl)-6,8-dimethylchroman-4-one and (3R)-5,7-dihydroxy-3-(4′-hydroxybenzyl)-6-methylchroman-4-one or isomer, respectively. In conclusion, these methods can be applied to identify and distinguish the quality of PR with other original plants and provide novel ideas for evaluating herbal products used in TCM.     

Nutritional and phytochemical value of Brassica crops from the agri‐food perspective

Brassica foods are among the top 10 economic crops in the world (i.e. broccoli, kale, cauliflower and Chinese cabbage). These vegetables have been identified as important components of a healthy diet because of their high levels of nutrients and health‐promoting phytochemicals (i.e. phenolics, glucosinolates, vitamins and minerals). Epidemiological studies have shown that increased consumption of Brassica foods is strongly associated with a reduced risk of degenerative diseases, cancer, cardiovascular disease and immune dysfunction. Nevertheless, the nutritional content and profile in Brassica vegetables have been reported to vary considerably during the growth period due to agronomical factors including light, temperature, water availability and soil fertility among others. Moreover, the conditions of postharvest processing and cooking are also important factors on food quality. A better understanding of specific preharvest and postharvest conditions is essential to improve cultivars with value‐added nutritional quality. Thus, in this article are going to be addressed the effects of the most common crop management strategies and processes on the variation of nutritive compounds present within Brassica from the agri‐food perspective.     

Comparative analysis of quality and nutritional traits from Lonicera caerulea (Honeyberry) cultivars and other berries grown in Scotland

Lonicera caeruleabrix is a perennial shrub native to North America, Europe and Asia. It produces dark blue berries known as honeyberries or haskap berries which are produced commercially in several territories including Canada, Japan, Russia and Poland. Plants are suited to UK environments, but it is yet to be widely commercially developed in the UK. In the present work, quality and nutritional traits of six honeyberry cultivars grown in Scotland were compared with other commonly grown berry fruits (strawberry, raspberry, blueberry, blackberry, blackcurrant) to aid the identification of environmentally stable, high-quality honeyberry cultivars suitable for UK cultivation. Differences were observed in fruit quality variables (soluble solids, titratable acidity and Brix/acid ratios) between honeyberry cultivars. Three of six cultivars examined exhibited notable variation in soluble solids dependent on harvest year with ‘Aurora’ and ‘Strawberry Sensation’ having consistently high ^oBrix values. Titratable acidity exhibited cultivar differences and there was limited variation over harvest years. ‘Aurora’ exhibited consistently high ^oBrix/titratable acidity ratio reflected by high glucose and fructose content. Honeyberry fruit had good nutritional profile relative to other soft fruits with higher polyphenol and anthocyanin content than strawberry, blueberry, blackberry and raspberry, manifested in greater antioxidant capacity. The major anthocyanins in aqueous honeyberry fruit extracts were cyanidin, pelargonidin and peonidin glycosides. These findings indicate that L. caerulea represents a crop suitable for UK cultivation capable of producing high quality fruit with a valuable nutritional profile relative to other soft fruits. Cultivars exhibit significant differences in fruit quality and nutritional profile as well as harvest consistency and growers should consider this when establishing new plantations.     

Analysis of genetic diversity and relationships in East African banana germplasm

The genetic diversity and phylogenetic relationships of 29 East African highland banana (Musa spp.) cultivars and two outgroup taxa, M. acuminata Calcutta 4 and Agbagba were surveyed by RAPD analysis. A genetic similarity matrix was established based on the presence or absence of polymorphic amplified fragments. Phylogenetic relationships were determined by UPGMA cluster analysis. RAPDs showed that the highland bananas are closely related with a narrow genetic base. Nevertheless, there were sufficient RAPD polymorphisms that were collectively useful in distinguishing the cultivars. The dendrogram was divisible into a major cluster composed of all the AAA highland banana cultivars and Agbagba (AAB) and a minor cluster consisting of Kisubi (AB), Kamaramasenge (AB) and Calcutta 4 (AA). Several subgroups are recognized within the major cluster. RAPD data did not separate beer and cooking banana cultivars. Our study showed that RAPD markers can readily dissect genetic differences between the closely related highland bananas and provide a basis for the selection of parents for improvement of this germplasm. Received: 28 June 2000 / Accepted: 1 August 2000     

种植模式对巴戟天生长的影响

为探讨种植模式对中药材巴戟天(Morinda officinalis)生长的影响,对两种种植模式下巴戟天生长性状进行了研究。结果表明,坡地连片种植基地(PLZ)的土壤有效矿质营养和有机质含量分别低于坡地围林种植基地(PWZ)的24%～28%和38%～43%,差异极显著(P 0.01)。PWZ种植3年的巴戟天茎、叶和肉质根形态指标均显著优于PLZ的(P 0.01);但随着栽培时间的延长,PLZ的巴戟天肉质根性状逐渐优于PWZ的,PLZ的6 a生巴戟天单株产量是PWZ的1.16～1.17倍;肉质根多糖、总黄酮和原花青素含量均显著高于PWZ的(P0.01),且有效成分含量均随种植时间的延长呈上升趋势(P0.01)。可见,巴戟天在PLZ模式下的肉质根质量好,入药更符合国家药典优品性状的要求。同时,建议合理增加肉桂围林种植基地的光照量,抑制藤苗生长,以提高巴戟天肉质根的产量和有效成分积累。     

Effects of Different Selenium Application Methods on Wheat (<i>Triticum aestivum</i> L.) Biofortification and Nutritional Quality

Mineral nutrient malnutrition, especially deficiency in selenium (Se), affects the health of approximately 1 billion people worldwide. Wheat, a staple food crop, plays an important role in producing Se-enriched foodstuffs to increase the Se intake of humans. This study aimed to evaluate the effects of different Se application methods on grain yield and nutritional quality, grain Se absorption and accumulation, as well as 14 other trace elements concentrations in wheat grains. A sand culture experiment was conducted via a completely randomized 3 × 2 × 1 factorial scheme (three Se levels × two methods of Se application, foliar or soil × one Se sources, selenite), with two wheat cultivars (Guizi No.1, Chinese Spring). The results showed that both foliar Se and soil Se application methods had effects on wheat pollination. Foliar Se application resulted in early flowering of wheat, while soil Se application caused early flowering of wheat at low Se levels (5 mg kg⁻¹ ) and delayed wheat flowering at high selenium levels (10 mg kg⁻¹ ), respectively. For trace elements, human essential trace elements (Fe, Zn, Mn, Cu, Cr, Mo, Co and Ni) concentrations in wheat grains were dependent of Se application methods and wheat cultivars. However, toxic trace elements (Cd, Pb, Hg, As, Li and Al) concentrations can be decreased by both methods, indicating a possible antagonistic effect. Moreover, both methods increased Se concentrations, and improved grain yield and nutritional quality, while the foliar application was better than soil. Accordingly, this study provided useful information concerning nutritional biofortification of wheat, indicating that it is feasible to apply Se to conduct Se biofortification, inhibit the heavy metal elements concentrations and improve yield and quality in crops, which caused human health benefits.     

Genomic cloning and linkage mapping of the Mal d 1 (PR-10) gene family in apple (Malus domestica)

Fresh apples can cause birch pollen-related food allergy in northern and central European populations, primarily because of the presence of Mal d 1, the major apple allergen that is cross-reactive to the homologous and sensitizing allergen Bet v 1 from birch. Apple cultivars differ significantly in their allergenicity. Knowledge of the genetic basis of these differences would direct breeding for hypoallergenic cultivars. The PCR genomic cloning and sequencing were performed on two cultivars, Prima and Fiesta, which resulted in 37 different Mal d 1 gDNA sequences. Based on the mapping of sequence-specific molecular markers, these sequences appeared to represent 18 Mal d 1 genes. Sixteen genes were located in two clusters, one cluster with seven genes on linkage group (LG) 13, and the other cluster with nine genes on the homoeologous LG 16. One gene was mapped on LG 6, and one remained unmapped. According to sequence identity, these 18 genes could be subdivided into four subfamilies. Subfamilies I–III had an intron of different size that was subfamily and gene-specific. Subfamily IV consisted of 11 intronless genes. The deduced amino acid sequence identity varied from 65% to 81% among subfamilies, from 82% to 100% among genes within a subfamily, and from 97.5% to 100% among alleles of one gene. This study provides a better understanding of the genetics of Mal d 1 and the basis for further research on the occurrence of allelic diversity among cultivars in relation to allergenicity and their biological functions.