Identification and genetic relationships of kenaf (Hibiscus cannabinus L.) germplasm revealed by AFLP analysis

Kenaf (Hibiscus cannabinus L.) is one of the world's most economically important fiber crops. In order to identify different varieties, and investigate its diversity and genetic relationships, twenty-three kenaf accessions and two accessions of its relative, roselle (H. sabdariffa var. altissima), were analyzed by morphological characterization and AFLP fingerprinting. It is very difficult to identify kenaf accessions based merely on morphological characters, due to their limited variation. For the AFLP study, a total of 505 polymorphic markers (out of 560) were produced by six selected AFLP primer combinations. The AFLP fingerprinting was effective in identifying all kenaf accessions included in the study. Kenaf and roselle are independent species with close relationships, and great genetic diversity was also detected among the kenaf accessions with different origins, based on the analysis of the AFLP markers. The AFLP analysis strongly supports the opinion that kenaf originated in Africa. It also demonstrated that the dissemination of kenaf was from Africa through Asia to Central and North America.     

AFLP Fingerprinting in Pigeonpea (Cajanus cajan (L.) Millsp.) and its Wild Relatives

Detection of DNA polymorphism in cultivated pigeonpea (Cajanus cajan) and two of its wild relatives Cajanus volubilis and Rhynchosia bracteata is reported here for the first time using amplified fragment length polymorphism (AFLP) fingerprinting. For this purpose, two EcoRI (three selective nucleotides) and 14 MseI (three selective nucleotides) primers were used. The two wild species shared only 7.15% bands with the pigeonpea cultivars, whereas 86.71% common bands were seen among cultivars. Similarly, 62.08% bands were polymorphic between C. volubilis and pigeonpea cultivars in comparison to 63.33% polymorphic bands between R. bracteata and pigeonpea cultivars, and 13.28% polymorphic bands among pigeonpea cultivars. The cluster analysis revealed low polymorphism among pigeonpea cultivars and very high polymorphism between cultivated pigeonpea and its wild relatives. The AFLP analysis also indicated that only one primer combination (EcoRI + ACT and MseI + CTG), at the most any four primer pair combinations, are sufficient for obtaining reliable estimation of genetic diversity in closely related cultivars like pigeonpea material analyzed herein. AFLP analysis may prove to be a useful tool for molecular characterization of pigeonpea cultivars and its wild relatives and for possible use in genome mapping.     

Identification of AFLP Markers that Discriminate Between Cultivated Cotton and the Hawaiian Island Endemic, <Emphasis Type="Italic">Gossypium tomentosum</Emphasis>Nuttall ex Seeman

Interspecific gene flow from crop species into wild relatives may compromise the genetic integrity of native species and in the case of transgenic crops lead to the escape of transgenes into natural populations. Approximately 72% of the upland cotton (Gossypium hirsutum L.) acreage grown in the United States in 2000 utilized transgenic cultivars. The closest relative of G. hirsutum is G. tomentosum Nuttall ex Seeman, endemic to the Hawaiian archipelago. Because these two species are fully interfertile, cultivation of transgenic G. hirsutum in the Hawaiian Islands is restricted. We investigated the possibility of identifying AFLP genetic markers that are diagnostic for each species and thus could be used in future studies to detect introgression between them. In addition, we were interested in comparing levels and geographic patterns of AFLP diversity in G. tomentosum to previous estimates using allozyme data. AFLP analysis led to the detection of 11 and 16 species-specific markers for G. tomentosum and G. hirsutum, respectively. These species-specific AFLP markers will be useful for detecting gene flow between G. hirsutum and G. tomentosum that has occurred in the past and thus might occur in the future if the restrictions on cultivation of transgenic G. hirsutum are relaxed in the Hawaiian Islands. Little genetic diversity and limited geographic patterning were discovered using AFLP markers, consistent with data from previous allozyme studies.     

Analysis of Characters and Germplasm of Significance to Improvement of Australian Native Waratahs (Telopea spp., family Proteaceae) for Cut Flower Production

Waratahs (Telopea spp.) are cultivated for their blooms for the international cut flower market. A morphometric study was conducted on a range of cultivated waratah varieties to determine the variability of selected characters of horticultural importance and which parents might be of value in future breeding programs. Univariate analysis of characters of 13 cultivars revealed the greatest range of variation in number of flowers, bract dimensions, flower colour and leaf margin type. Differences were observed between number of flowers per inflorescence in the T. speciosissima (Sm.) R.Br. group of accessions and the other species cultivars; interspecific cultivars with T. mongaensis, Cheel and T. oreades F. Muell. were intermediate, with a similar pattern observed in bract length and width. Multivariate analysis (canonical variate analysis – CVA) discriminated between waratah cultivars on the basis of flower colour (first axis), then flower number, bract length, leaf width and leaf margin type (axis 2). The third axis also separated leaf width, the fourth leaf length and width, and the fifth leaf apex shape, CVA was also used to explore the genetic contribution of three parent T. speciosissima cultivars to hybrid populations. Hybrids with ‘Sunflare’, ‘Sunburst’ or ‘Wirrimbirra White’ as one parent were very dispersed indicating the range of inheritance of the observed phenotypic characters, leading to the possibility of selecting individuals with the required degree of character inheritance. It was demonstrated that the Telopea speciosissima type, which forms the basis of the waratah cut flower industry, may be improved by hybridising with other Telopea species. The application of the results to the development of waratah breeding programs is discussed.     

Identification of powdery mildew and leaf rust resistance genes in common wheat (Triticum aestivum L.). Wheat varieties from the Caucasus, Central and Inner Asia

225 wheat varieties from Kazakhstan, Mongolia and Russia were analysed for their resistance to powdery mildew and leaf rust diseases. A set of 11 different Blumeria graminis f. sp. tritici isolates was used to test for powdery mildew resistance and a set of 10 different Puccinia triticina isolates for leaf rust. 115 cultivars/lines were susceptible to powdery mildew and 32 cultivars/lines showed an intermediate resistance response. 21 cultivars/lines revealed the response pattern of individual resistance genes Pm1, Pm2, Pm3, pm5, Pm6, Pm8, Pm9, Pm17 and Pm22, respectively, therefrom three line showed a combination of two resistance genes and two varieties a combination of three genes. 50 cultivars/lines showed resistance to some specific isolates but an assignment to known resistance genes or gene combinations was not possible, whereas seven lines were completely resistant to all used isolates. The leaf rust test showed that 83 cultivars/lines were susceptible and 11 lines revealed intermediate resistance response. 62 cultivars/lines could be assumed to possess major resistance genes Lr1, Lr3, Lr10, Lr23 and Lr26, respectively, therefrom seven cultivars possessed a combination of two resistance genes. Lr3 was the most widespread resistance gene, occurring in 42 cultivars/lines. 13 lines were completely resistant to all used isolates, however, the response patterns of 56 cultivars/lines did not match to any known gene or gene combination. In 13 varieties the T1BL·1RS wheat-rye translocation could be identified, in five cultivars resistance gene Pm8 was suppressed.     

Biological nitrogen fixation in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) by <Superscript>15</Superscript>N isotope-dilution and identification of associated culturable diazotrophs

The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the ¹⁵N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.     

Genetic Relationship and Diversity of Four MangiferaSpecies Revealed through AFLP Analysis

We used AFLP analysis to explore the genetic relationship and diversity between and within 4 Mangifera species. We analyzed 35 accessions comprising 8 cultivars and 3 landraces of M. indica L., 11 landraces of M. odorata Griff., 7 landraces of M. foetida Lour., and 6 landraces of M. caesia Jack. Using 8 primer combinations produced a total of 518 bands, 499 (96.3%) of which were polymorphic among the 35 accessions. Clustering analysis showed that all 35 accessions were basically classified into 4 groups corresponding to the 4 Mangifera species. Our results indicate that the genetic relationship of these 4 Mangifera species based on AFLP analysis is in good agreement with their classification by classic methods. In addition, it was clearly revealed the genetic diversity between and within 4 Mangifera species. The findings obtained in this study are useful for the breeding in Mangifera species.     

Assessment of genetic relationships among Pyrus species and cultivars using AFLP and RAPD markers

Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.     

Genetic Variation and Relationships of Pedigree-Known Oat, Wheat, and Barley Cultivars Releaved by Bulking and Single-Plant Sampling

Applications of bulking procedures have played an increasingly important role in molecular characterization of plant germplasm, but little attention has been made to address the effectiveness of detecting genetic variation and inferring genetic relationships via bulking. An analysis was performed here to compare the genetic variation detected and genetic relationships inferred via bulking and single-plant sampling of five oat (Avena sativa L.), wheat (Triticum aestivum L.), and barley (Hordeum vulgare L.) cultivars with known pedigrees using amplified fragment length polymorphism (AFLP) markers. Three AFLP primer pairs were applied to screen one bulk and eight single-plant samples of each cultivar and up to 140 AFLP bands were scored for each sample. Analyses of these AFLP data showed bulking revealed AFLP variation up to 21.4% less than corresponding single-plant sampling for these crop species and also introduced up to 2.2% upward and 5.1% downward biases in detecting AFLP variations for each cultivar. The genetic relationships inferred by bulking using the Dice's coefficient, the simple matching coefficient, and the Jaccard's coefficient were largely the same, but differed from those in single-plant samples employing average Dice's coefficient, average simple matching coefficient, and AMOVA-based distance method. All of the inferred genetic relationships were not congruent to the known pedigrees. Clearly, substantial biases could exist in detection of AFLP variation and in inference of genetic relationships from bulk samples, even for closely related germplasm, and more efforts to assess the effectiveness of bulking in inferring genetic variation and relationships are needed for more informative molecular characterization of plant germplasm.     

Relationship of European persimmon (Diospyros kaki Thunb.) cultivars to Asian cultivars, characterized using AFLPs

Sixty one persimmon (Diospyros kaki Thunb.) selections, including 17 Italian, 11 Spanish, 13 Japanese, six Korean, five Chinese, one Israeli, and eight of unknown origin, were evaluated for genetic differences by AFLP analysis. Relationships among cultivars were evaluated by UPGMA clustering, Neighbor Joining, and MultiDimensional Scaling. While similarities among groups were generally less than 0.60, both UPGMA and Neighbor Joining separated European and Asian cultivars. Spanish and Italian cultivars were not separated by any of the analyses, suggesting that they share a common gene pool, while Japanese, Chinese and Korean cultivars formed distinct clusters. Diversity within groups was greater than diversity between groups. Most cultivars were quite polymorphic (only 0.60–0.80 similarity between cultivars). In addition, the presence of several Japanese cultivars in the European group and a group of European cultivars nested between Chinese and Korean groups suggest that similar, but different progenitors were used in the development of the present European cultivars. ‘Kaki Tipo’ selections from different sources were clearly different by AFLP analysis, indicating that they are separate cultivars.     

Assessment of genetic diversity in cultivars and wild accessions of Luohanguo (<Emphasis Type="Italic">Siraitia grosvenorii</Emphasis> [Swingle] A. M. Lu et Z. Y. Zhang), a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers

Genetic variation of wild populations and cultivars of Luohanguo (Siraitia grosvenorii), a plant species endemic to southern China, was assessed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. Based on the results for 130 individuals from seven populations, a high level of genetic diversity of Luohanguo was observed at the species level. The percentage of polymorphic loci (P) was 89.4%, Nei’s gene diversity (H _e) was 0.239, and Shannon’s information index (H _o) was 0.373 based on the combined AFLP and RAPD data. There was a high degree of genetic differentiation, with 45.1% of the genetic variation attributed to differences between the populations. The genetic diversity of the Luohanguo cultivars is much lower than that of wild populations (P = 41.8%, H _e = 0.141, H _o = 0.211), and a distinct genetic differentiation is observed between the cultivars and wild accessions. The pool of genetic variation in the wild populations provides an excellent gene resource for Luohanguo breeding.     

Genetic Diversity among Syrian Cultivated and Landraces Wheat Revealed by AFLP Markers

Genetic diversity among some important Syrian wheat cultivars was estimated using Amplified Fragment Length Polymorphism (AFLP) markers. Five Triticum aestivum L. and 10 Triticum turgidum ssp. durum were analyzed with 11 EcoRI–MseI primer pair combinations. Of the approximately 525 detected AFLP markers, only 46.67% were polymorphic. Cluster analysis with the entire AFLP data divided all cultivars into two major groups reflecting their origins. The first one contained T. aestivum L. cultivars, and the T. turgidum ssp. durum cultivars and landraces were grouped in the second. Narrow genetic diversity among all cultivars was detected with an average genetic similarity of 0.884. The lowest similarity index (0.9) was found between Cham5 and Hamary (durum wheat), whereas this value was 0.93 between Salamony and Bouhouth 4 (T. aestivum L.). The narrow genetic diversity level indicates that these genotypes could be originated from the same source. AFLP analysis provides crucial information for studying genetic variation among wheat cultivars and provides important information for plant improvement.     

Identification and evaluation of <Emphasis Type="Italic">Forsythia</Emphasis> germplasm using molecular markers

This study identifies Forsythia germplasm and evaluated the genetic relationships of F. ×intermedia hybrids, cultivars and their putative parental species. Leaf samples of F. ×intermedia cultivars and species, such as F. koreana and F. suspensa, were collected in the Netherlands, Korea, and USA. Total genomic DNA was extracted and evaluated by randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses. Dendrograms were constructed by using the neighbor-joining (NJ) clustering algorithm applying the interior branch (IB) test or analyzed by STRUCTURE. In the dendrogram generated by RAPD markers, two major clusters were observed. One cluster (CL-I) contained most of the F. ×intermedia cultivars, F. suspensa, and F. koreana. The other cluster (CL-II) included F. europaea, F. ovata, F. densiflora, F. mandshurica, F. japonica, F. viridissima, and cultivars derived from F. ovata. In the AFLP dendrogram, the placement of F. ×intermedia cultivars with F. suspensa was similar, forming cluster CL-A I. The RAPD and AFLP results clearly separated most F. ×intermedia cultivars from F. ovata derived cultivars. The full range of genetic diversity of F. suspensa and F. viridissima should be investigated to verify whether these two species are truly parental taxa for F. ×intermedia. Placement of F. viridissima, F. ovata, and F. japonica in different sub-clusters requires further investigation regarding genetic diversity in the species, and their close relationship with F. koreana, F. mandshurica, and F. saxatalis.     

Phylogenetic relationships within <Emphasis Type="Italic">Citrus</Emphasis> and its related genera as inferred from AFLP markers

We investigated the phylogenetic relationships among Citrus and its relatives, including 29 genotypes belonging to Citrus, Poncirus, Fortunella, Microcitrus, Eremocitrus, Atalantia and Severinia using AFLP analysis. Cluster analysis was conducted using neighbour joining (NJ) method. The results demonstrated that Poncirus, Microcitrus and Eremocitrus are distant from Citrus. A strong affiliation exists between C. halimii and Fortunella and the results did not support C. halimii as the fourth basic species. Further evidence was provided that P. polyandra should not be the hybrid between trifoliate orange and Citrus genotypes and it deserves species status. Neither the separation of subgenus Papeda and Citrus nor the separation of subgenus Archicitrus and Metacitrus were clearly resolved in the dendrogram. C. ichangesis is a distinct species very different from other Citrus genotypes, and the results showed it is improper to classify it into subgenus Metacitrus. C. reticulata, C. maxima and C. medica were separated into three distinct clusters. Therefore if only cultivated Citrus species are considered, the three basic species concept is acceptable. However, it is not applicable for whole genus Citrus.     

Assessment of oil content and fatty acid composition variability in two economically important Hibiscus species

The Hibiscus genus encompasses more than 300 species, but kenaf (Hibiscus cannabinus L.) and roselle (Hibiscus sabdariffa L.) are the two most economically important species within the genus. Seeds from these two Hibiscus species contain a relatively high amount of oil with two unusual fatty acids: dihydrosterculic and vernolic acids. The fatty acid composition in the oil can directly affect oil quality and its utilization. However, the variability in oil content and fatty acid composition for these two species is unclear. For these two species, 329 available accessions were acquired from the USDA germplasm collection. Their oil content and fatty acid composition were determined by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Using NMR and GC analyses, we found that Hibiscus seeds on average contained 18% oil and seed oil was composed of six major fatty acids (each >1%) and seven minor fatty acids (each <1%). Hibiscus cannabinus seeds contained significantly higher amounts of oil (18.14%), palmitic (20.75%), oleic (28.91%), vernolic acids (VA, 4.16%), and significantly lower amounts of stearic (3.96%), linoleic (39.49%), and dihydrosterculic acids (DHSA, 1.08%) than H. sabdariffa seeds (17.35%, 18.52%, 25.16%, 3.52%, 4.31%, 44.72%, and 1.57%, respectively). For edible oils, a higher oleic/linoleic (O/L) ratio and lower level of DHSA are preferred, and for industrial oils a high level of VA is preferred. Our results indicate that seeds from H. cannabinus may be of higher quality than H. sabdariffa seeds for these reasons. Significant variability in oil content and major fatty acids was also detected within both species. The variability in oil content and fatty acid composition revealed from this study will be useful for exploring seed utilization and developing new cultivars in these Hibiscus species.     

Genetic Variation of Ethiopian Mustard (Brassica carinata A. Braun) Germplasm in Western Canada

Information on genetic diversity and genetic relationships among genotypes of Brassica carinata is currently limited. The objectives of this study were to evaluate patterns and levels of genetic diversity in B. carinata based on amplified fragment length polymorphisms (AFLP) as compared with Brassica juncea and Brassica nigra, and to evaluate agronomic and seed quality data for plants grown in the field in western Canada. A total of 296 AFLP bands were generated from four primer pair combinations and scored for presence/absence in 66, 20 and 7 accessions of B. carinata, B. juncea and B. nigra, respectively. B. carinata was less genetically diverse than the other two species. Differences in diversity were evident in the proportion of polymorphic loci within each species: 23, 35 and 50% for B. carinata, B. nigra and B. juncea, respectively. Pair-wise similarity measures based on the Jaccard coefficient were highest among accessions of B. carinata and showed the narrowest range: 0.911 (0.810–0.981) compared to B. nigra: 0.569 (0.438–0.660) and B. juncea: 0.715 (0.345–0.951). AFLP-based genetic distance information can be used by plant breeders to select diverse genotypes. AFLPs are also useful for fingerprinting cultivars and two primer pair combinations were sufficient to uniquely identify all the accessions of B. carinata. More variation among accessions was identified in the agronomic trial than had previously been described in studies of B. carinata in western Canada, but the data were too limited to draw conclusions regarding specific accessions. Overall, the findings were in agreement with other published work describing the favourable agronomic potential of this species.     

Genetic Diversity of the Greater Yam (<Emphasis Type="Italic">Dioscorea alata</Emphasis> L.) and Relatedness to <Emphasis Type="Italic">D. nummularia</Emphasis> Lam. and <Emphasis Type="Italic">D. transversa</Emphasis> Br. as Revealed with AFLP Markers

Amplified fragment length polymorphism markers were used to assess the genetic relatedness between Dioscorea alata and nine other edible Dioscorea. These species include D. abyssinica Hoch., D. bulbifera L., D. cayenensis-rotundata Lamk. et Poir., D. esculenta Burk., D. nummularia Lam., D. pentaphylla L., D. persimilis Prain. et Burk., D. transversa Br. and D. trifida L. Four successive studies were conducted with emphasis on the genetic relationship within D. alata and among species of the Enantiophyllum section from Vanuatu. Study 1 was carried out to select a set of polymorphic primer pairs using 11 combinations and eight species belonging to five distinct sections. The four most polymorphic primer pairs were used in study 2 among six species of the Enantiophyllum section. Study 3 focussed mainly on the genetic relationship among 83 accessions of D. alata, mostly from Vanuatu (78 acc.) but also from Benin, Guadeloupe, New Caledonia and Vietnam. The ploidy level of 53 accessions was determined and results indicated the presence of tetraploid, hexaploid and octoploid cultivars. Study 4, included 35 accessions of D. alata, D. nummularia and D. transversa and was conducted using two primer pairs to verify the taxonomical identity of the cultivars `langlang', `maro' and `netsar' from Vanuatu. The overall results indicated that each accession can be fingerprinted uniquely with AFLP. D. alata is an heterogeneous species which shares a common genetic background with D. nummularia and `langlang', `maro' and `netsar'. UPGMA cluster analysis revealed the existence of three major groups of genotypes within D. alata, each assembling accessions from distant geographical origins and different ploidy levels. The analysis also revealed that `langlang', `maro' and `netsar' clustered together with the cultivar `wael' (D. transversa) from New Caledonia. Results are discussed in the paper.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.