A preliminary study of the genetic diversity of Bolivian oca (<Emphasis Type="Italic">Oxalis tuberosa</Emphasis> Mol.) varieties maintained<Emphasis Type="Italic"> in situ</Emphasis> and<Emphasis Type="Italic"> ex situ</Emphasis> through the utilization of ISSR molecular markers

ISSR molecular markers have been used to investigate genetic diversity of oca (Oxalis tuberosa Mol.), an Andean neglected tuber crop species. Sampling procedure allowed a preliminary study of the genetic diversity at the intra- and intervarietal levels. Twenty tuber lots conserved in situ in the microcentre of Candelaria and ex situ in the Toralapa Centre (Bolivia) were identified. Four ISSR primers amplified a total of 25 fragments of which 17 (68%) were polymorphic. These experiments show that the structure of oca varieties is mainly based upon vernacular names with a greater differentiation among tuber lots than within them, supporting agromorphological data. ISSR technique enlightened the existence of heterogeneous varieties in oca and divergence between in situ and ex situ conservation strategies. These observations are potentially linked to the different ways of management of tubers in these two conservation systems.     

Conservation in a changing world: <Emphasis Type="Italic">in situ</Emphasis> conservation of the giant maize of Jala

Using survey data from Jala, Mexico, this case-study evaluates in situ maize conservation of the variety ‘Jala’ (Zea mays L.). Though historically ‘Jala’ was the dominant variety grown in the valley of Jala, today less than 20% of farmers grow it on only 5% of the maize area. Younger growers of the ‘Jala’ variety specialize in it, growing relatively large amounts for niche markets. Older, diversified farmers grow small areas for household use and to compete in a local contest. Conservation of the ‘Jala’ variety has been heavily influenced by shifting ideal concepts of maize, as determined by market and consumption demands and by a contest designed to promote in situ conservation. The current move away from nationalized purchasing may favor ‘Jala’s’ continued conservation.     

Capturing genetic diversity of wild populations for <Emphasis Type="Italic">ex situ</Emphasis> conservation: Texas wild rice (<Emphasis Type="Italic">Zizania texana</Emphasis>) as a model

Genebanks complement other conservation programs because they preserve genetic diversity needed for future breeding and restoration. We evaluated efficiency of capturing genetic diversity, using endangered Zizania texana (Texas wild rice) as a model for plants with recalcitrant seeds. This perennial aquatic grass is restricted to 4 km of the San Marcos River in Texas. An early conservation collection included plants from stands throughout the river, based on the assumption stands would be unique genotypes. Using microsatellite markers, we found that genetic diversity was concentrated in five of 15 large, demographically stable stands; 96 stands smaller than 2 m² contributed no unique alleles. High heterozygosity and few duplicate genotypes suggested that sexual reproduction occurs more often than presumed. Simulations of stratified sampling of large stands captured all alleles in only 45 individuals, while random sampling along the river captured much less diversity. The early conservation collection captured as much diversity as expected from random sampling. Texas wild rice stands resemble a mainland-island metapopulation; our analyses suggest that stratified sampling maximizes genetic diversity for this population dynamic. Demographic and genetic information is important for validating the design of efficient ex situ collections and guiding in situ conservation.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work     

Aspects of the evolution of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. and the status of the United States <Emphasis Type="Italic">Nicotiana</Emphasis> Germplasm Collection

The genus Nicotiana is a member of the nightshade (Solanaceae) family, and is comprised of 70 currently recognized, naturally occurring species. Genetic variability within N. tabacum L., the species of primary economic importance, was likely affected by several genetic bottlenecks. Nicotiana tabacum is a classic amphidiploid that arose after chance interspecific hybridization between N. sylvestris Spegazinni et Comes and a member of section Tomentosae, likely N. tomentosiformis Goodspeed, N. otophora Grisebach, or an introgressive hybrid between the two. Only a fraction of the genetic variability that existed in the diploid progenitor gene pools probably entered into N. tabacum. Genetic drift, coupled with natural and human selection, subsequently resulted in the formation of narrow genetic pools corresponding to modern commercial market classes. Genetic variability in Nicotiana has gained increased attention in recent years because of investment in Nicotiana genomics research, interest in development of tobacco products with reduced harm characteristics, and concentration on using Nicotiana species for plant-based production of commercially useful proteins. A storehouse of genetic diversity for N. tabacum is available in approximately 1,900 accessions maintained by the United States Nicotiana Germplasm Collection. Seeds of 224 accessions representing 59 wild Nicotiana species are also maintained. The collection is currently maintained by North Carolina State University and is part of the United States National Plant Germplasm System (NPGS). The collection’s curator satisfies hundreds of seed requests made annually by scientists using Nicotiana germplasm for basic biological investigations and by researchers in the area of applied tobacco science.     

The Corsican citron melon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis> (Thunb.) Matsum. et Nakai subsp. <Emphasis Type="Italic">lanatus</Emphasis> var. <Emphasis Type="Italic">citroides</Emphasis> (Bailey) Mansf. ex Greb.) a traditional and neglected crop

During a collecting mission in Corsica (France) three landraces of citron melon (Citrullus lanatus (Thunb.) Matsum. et Nakai subsp. lanatus var. citroides (Bailey) Mansf. ex Greb.) were collected in the northern areas. Today in Corsica this old and neglected crop is in decline and risks extinction. A strategy for its characterization and safeguarding is in progress at the genebank of IGV of Bari (Italy).     

Evaluation of perennial wild <Emphasis Type="Italic">Cicer</Emphasis> species for drought resistance

About 90% of chickpea (Cicer arietinum L.) in the world is grown under rainfed conditions where drought is one of the major constraints limiting its productivity. Unlike the cultivated chickpea, wild Cicer species possesses sources of resistance to multiple stresses; we therefore evaluated perennial wild Cicer species for resistance to drought. C. anatolicum, C. microphyllum, C. montbretii, C. oxydon and C. songaricum were compared with special checks; C. echinospermum, C. pinnatifidum and C. reticulatum and five cultivated chickpeas. After the cultivated chickpeas were killed, accessions were evaluated using a 1–5 scale, where 1 = highly drought resistant (no visible drought effect and full recovery after three successive wiltings) and 5 = highly drought susceptible (leaves and branches dried out, no recovery at all). All accessions of perennial wild Cicer species were significantly superior to those annual wild species and the cultivated chickpeas including the best drought tolerant chickpea, ICC 4958 under drought conditions. Perennial wild Cicer species did not only recover after wilting and drying out above ground level, they also tolerated high temperatures up to 41.8°C. But, they do not cross with the cultivated chickpeas. C. anatolicum should be taken account in long term breeding programs because it has closer affinities to the first crossability group than the others.     

Diversity of aroma patterns in wild and cultivated <Emphasis Type="Italic">Fragaria</Emphasis> accessions

Plant breeders are interested in strawberry species as donors of volatile compounds in breeding programmes because of the diversity and intensity of wild strawberry aroma. Therefore, the topic of this paper is the prospective analysis of four accessions of four wild strawberry accessions in comparison to a standard cultivar of Fragaria × ananassa Duch. by using human sensory, gas chromatography mass spectrometry (GCMS) and gas chromatography–olfactometry (GCO). The wild species have higher aroma intensities compared with the cultivated one. The flavour quality differs significantly. Semiquantitative GC analysis revealed that F. × ananassa cv. ‘Elsanta’ has the lowest content of volatile compounds whereas Fragaria moschata L. ‘Cotta’ has the highest. The aroma impressions, measured by GCO, support the findings of GCMS analyses. The nasal impact frequency (NIF)-profiles of the wild types are more manifold and of higher intensities than those of the cultivated F. × ananassa cv. ‘Elsanta’ which corresponds with the overall flavour impression when tasting the fresh fruits.     

Characterization of intergeneric hybrids of <Emphasis Type="Italic">Erianthus rockii</Emphasis> and <Emphasis Type="Italic">Saccharum</Emphasis> using molecular markers

Erianthus rockii, a wild relative of sugarcane, is drought and cold tolerant, and both are potentially important agronomic traits to the sugarcane industry worldwide. As such it is of interest as a source of parental germplasm to sugarcane breeders and is currently being used in sugarcane introgression programs in both China and Australia. To date morphological characters have been used to verify the putative hybrids produced. Two crosses were generated between two different Saccharum species and two E. rockii accessions. Over 400 AFLP markers were used to identify the intergeneric hybrid progeny as well as determine hybrid diversity. Both crosses generated hybrids but efficiency levels were very different and are probably related to the different Saccharum parent used in each cross. Cross 1 was between a Saccharum officinarum and E. rockii and generated 100% hybrid progeny. Cross 2, however, was between a sugarcane hybrid (S. officinarum × Saccharum spontaneum) and E. rockii and only 10% of the progeny were intergeneric hybrids. Inheritance of markers in the progeny was analysed and for both crosses there were equal numbers of markers from both parents indicating n + n transmission of chromosomes. This is the first verification of E. rockii hybrids with molecular markers. It may now be possible to exploit genes of value from E. rockii in sugarcane breeding programs.     

Genetic diversity and conservation of two endangered eggplant relatives (<Emphasis Type="Italic">Solanum vespertilio</Emphasis> Aiton and <Emphasis Type="Italic">Solanum lidii</Emphasis> Sunding) endemic to the Canary Islands

Solanum vespertilio Aiton and Solanum lidii Sunding are endemic, endangered wild species from the Canary Islands. These species are of potential value for eggplant (S. melongena) breeding, given that they are part of the secondary genepool of this crop. We study genetic diversity with amplified fragment length polymorphisms (AFLPs) markers from 5 populations of S. vespertilio (47 samples) and 3 of S. lidii (26 samples). Five related African species (S. dasyphyllum Schumach. et Thonn., S. delagoense Dunal, S. campylacanthum Hochst., S. panduriforme E. Mey, S. aff. violaceum Ortega) were also included in the analysis. A total of 235 AFLP markers included 178 and 156 that were polymorphic in S. vespertilio and S. lidii, respectively. Analysis of genetic distance, phenograms, and principal component plots showed that these rare Canarian species are differentiated (G _ST = 0.412) from the continental materials and that Solanum vespertilio is more distinct to its African congeners than is S. lidii. There is a relatively high level of differentiation between the two species (G _ST = 0.373), that presumably reflects geographic restrictions (S. lidii to Gran Canaria; S. vespertilio essentially to Tenerife). However, both species have similar levels of total diversity. We speculate that the combination of the many unusual reproductive features (andromonoecy, zygomorphy, heteranthery and weak enantiostyly in S. vespertilio) help explain genetic diversity that is high for self compatible species. The high genetic diversity may also indicate populations were larger in the past. A decrease in population size could contribute to the relatively low genetic differentiation among the populations. The data presented herein provide the foundation for initiation of ex situ and in situ conservation programs for these wild relatives of eggplant. This paper is dedicated to the memory of Dr. Richard N. Lester, who made significant contributions to the taxonomy, biosystematics and conservation of genetic resources of African species of Solanum.     

Seed inoculation with <Emphasis Type="Italic">Bacillus</Emphasis> spp. improves seedling vigour in oil-seed plant <Emphasis Type="Italic">Jatropha curcas</Emphasis> L.

Jatropha (Jatropha curcas L.) is a non-edible oil-seed plant with adaptability to marginal semi-arid lands and wastelands. The Indian Government is promoting jatropha to reduce dependence on the crude oil and to achieve energy independence by the year 2012, under the National Biodiesel Mission. Selected strains of Bacillus spp., either supplemented with or without chitin, were tested for their ability to promote growth of jatropha seedlings in pot culture studies. The strains supported growth of jatropha seedlings up to 42 days after sowing. Among all strains, Bacillus pumilus (IM-3) supplemented with chitin showed over all plant growth promotion effect resulting in enhanced shoot length (113%), dry shoot mass (360%), dry root mass (467%), dry total plant mass (346%), leaf area (256%), and chlorophyll content (74%) over control. Treating seeds with strain IM-3 without chitin resulted in enhanced dry shoot mass (473%), dry total plant mass (407%), and chlorophyll content (82%). However, Bacillus polymyxa (KRU-22) with chitin supported maximum root length (143%). Either strain IM-3 alone or in combination with other promising strains could be promoted further for enhanced initial seedling growth of jatropha.     

Characterization of Portuguese populations of <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. ssp. <Emphasis Type="Italic">sylvestris</Emphasis> (Gmelin) Hegi

Wild populations of Vitis vinifera L.␣have been located in Portugal. Morphological characterization was carried out in three populations located in Alcácer do Sal, Castelo Branco, and Montemor-o-Novo, and then compared using multivariate discriminant analysis. These populations were from three different hydrological basins, therefore cross-pollination was not possible. It was verified that in each population all plants were different. The data suggest that the frequency of female and male plants is rather variable in wild populations. The morphology of the adult leaf, from the Alcácer do Sal population, had particular features when compared with Castelo Branco and Montemor-o-Novo populations, which were more homogeneous. The length of teeth compared with width at the end of the base, and the density of prostrate hairs between and on main veins (lower side) were the variables which allowed the best discrimination among populations.     

Analyses of the seed protein contents on the cultivated and wild buckwheat <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> resources

The seed protein contents of 179 accessions belonging to ten species of genus Fagopyrum were determined by means of the Kjeldahl method. The results indicated that there are significant differences of seed protein content among different accessions within the same species. The average protein content of F. esculentum is 12.94%, ranging from 8.81–18.71%, and the protein content of Sibano is the highest, up to 18.71%, Gantian 1 the lowest with 8.81%. The average protein content of tartary buckwheat is 12.17%, with the range from 7.82–18.94%, and the protein content of cultivated tartary buckwheat Qianku 2 is the highest (18.59%), and that of Ganku 1 (7.82%) the lowest. Moreover, there are significant differences of seed protein content among different buckwheat species and between the different buckwheat types (between common buckwheat and tartary buckwheat, between the diploid and the tetraploid, between the cultivated and the wild, and between the big-achene group and the small-achene group). The average seed protein content of F. giganteum is the highest in all tested species, up to 17.81% with the range of 13.91–21.27%, and that of F. callianthum the lowest, only 8.31%. This study was financed by the Natural Science Foundation of China (30270852, 30471116), Program for New Century Excellent Talents in University (NCET-2004-0913), Guizhou Key International Cooperation Project (Qianke Hewai G Zi 2005#400108), Guizhou Oversea Talent Project (Qianren Xiangmu Zizhu Hetong 2004#02), and Key Project of Students in Guizhou Normal University (GNU-SP-2005-SJXY-A01)     

Production of Intergeneric Hybrids Between <Emphasis Type="Italic">Sinapis alba</Emphasis> and <Emphasis Type="Italic">Brassica carinata</Emphasis>

Reciprocal crosses were made between Brassica carinata and its related species Sinapis alba. Pollen germination studies indicated the presence post-fertilization barriers in both ways. Sequential ovary–ovule culture helped to realize the intergeneric hybrids from the cross S. alba × B. carinata. The F₁ obtained was confirmed as a hybrid based on morphology, cytology and isozyme studies. The hybrid was backcrossed to its male parent and obtained BC₁ seeds, which were used to raise BC₁ generation. The BC₁ generation plants were further backcrossed to B. carinata in order to develop alloplasmic lines.     

Genetic diversity of <Emphasis Type="Italic">rbc</Emphasis>L gene in <Emphasis Type="Italic">Elymus trachycaulus</Emphasis> complex and their phylogenetic relationships to several Triticeae species

Elymus trachycaulus complex species are known for their morphological variability, but little is known about their genetic basis. The phylogenetic relationships among the E. trachycaulus complex, and their systematic relation to other species in Triticeae remain unknown. Nucleotide diversity of ribulose-1,5 bisphosphate-carboxylase (rbcL) gene in E. trachycaulus complex species and several other Triticeae was first characterized and compared. A primary conclusion of the present study is that nucleotide diversity for rbcL gene in E. trachycaulus species was detected with the estimates of nucleotide diversity θ = 0.00039 and π = 0.00043. The estimate of nucleotide diversity in rbcL gene for species with different genome constitution here ranged from 0.00099 (π) and 0.00099 (θ) for the species with Ns genome to 0.00226 (π) and 0.00291 (θ) for the species with St genome. The phylogenetic relationships of these species were assessed using these rbcL sequences. A total of 47 variable positions including 19 parsimony-informative sites were detected among 24 accessions of 18 species/subspecies. The species with St, H/I and Ns genomes well separated from each other, and formed a three distinct clades with higher bootstrap values support for both Parsimony and NJ analyses. The St genome containing species is sister group of H/I genome containing species. Our result confirms that Pseudoroegneria is the maternal genome donor to these Elymus species studied here, regardless of their distribution. Elymus trachycaulus complex are more related to each other than to E. glaucescens, E. patagonicus, and E. solandri. This study suggested that Elymus species with StH genomes may form from multiple closely related sets of donors.     

Isozyme Variation in <Emphasis Type="Italic">Passiflora</Emphasis> Subgenus <Emphasis Type="Italic">Tacsonia</Emphasis>: Geographic and Interspecific Differentiation Among the Three Most Common Species

In a general study of banana passion fruit genetic resources, diversity was analyzed in the two main cultigens, P. tripartita var. mollissima and P. tarminiana, and their closest wild relative, P. mixta, scoring isozyme bands (IDH,PGM,ACP,PGD,DIA,andPRX) on288 plants from 31 accessions. Polymorphismandallelic richness, Nei diversity indices, and neighbor joining clustering showed that variation was poor in the cultigens in northern and central Colombia, while P. mixta appeared much more polymorphic. The populations of P. tripartita var. mollissima and P. mixta from southern Colombia and Ecuador show higher diversity values and are clearly differentiated from those of central and northern Colombia. This geographic component of variation is even stronger than the interspecific one, which suggests a close relation and a regular gene flow between these two species. In contrast, all the accessions of P. tarminiana constitute a clearly differentiated group, even if some introgression with P. tripartita var. mollissima is also suspected. The high variation observed in the southern region indicates the proximity of a center of diversity for banana passion fruit and collecting in Ecuador, Peru and Bolivia is recommended. The high diversity of P. mixta and the evidence of gene flow with P. tripartita var. mollissima constitute a favorable context for the implementation of in situ conservation strategies.     

Acquisition and ecological characterization of <Emphasis Type="Italic">Lactuca serriola</Emphasis> L. germplasm collected in the Czech Republic,Germany, the Netherlands and United Kingdom

Expeditions were conducted in the Czech Republic, Germany, the Netherlands and United Kingdom in 2001 (partly 1998) to study Lactuca serriola L. (prickly lettuce, compass plant) geographic distribution, ecology, habitat characteristics and occurrence of diseases and pests on this species. During these missions the seed material of L. serriola L. was collected in an east–west transect of these four countries. The European transect where seeds were collected and field observations were made represents a relatively large area between 2°34′50′′ W–17°32′46′′ E and 47°40′42′′–54°04′19′′ N. The seed material was used for regeneration, inclusion in the national genetic resources collections of individual countries and for research purposes in follow-up studies. During the missions, 50 locations with occurrence of L. serriola L. were visited (16 in Czech Republic and Germany, 10 in UK and 8 in the Netherlands). Individual seed lots of sixteen different plants were collected at each location (L. serriola L. population). Thus, in total 800 seed samples were collected. In Czech Republic and Germany L. serriola L. f. serriola dominated in all observed populations, in the Netherlands both f. serriola and f. integrifolia occurred in pure or mixed stands, whereas in the United Kingdom L. serriola L. f. integrifolia (S.F. Gray) S.D. Prince et R.N. Carter was dominant. L. serriola L. was recorded at various altitudes (−4 to 410 m), different habitats (ditches, along roads, fields and field margins, ruderal places, pavements and parking sites, fallow fields), individual populations varied substantially in size (20 to >1000 plants), area covered (25–10,000 m²), and the structure and character of associated plant communities. Natural infections by downy mildew (Bremia lactucae Regel) and powdery mildew (Golovinomyces cichoracearum (DC.) V.P. Gelyuta) were observed in some populations. B. lactucae Regel was recorded only in Czech Republic, G. cichoracearum (DC.) V.P. Gelyuta was more common in continental Europe. General and specific aspects of L. serriola L. geographic distribution and ecology are discussed.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.