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.     

AFLP similarities among historic Danish cultivars of fodder beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L. subsp. <Emphasis Type="Italic">vulgaris</Emphasis>var. <Emphasis Type="Italic">rapacea</Emphasis> Koch)

Dead seeds of a fodder beet cultivar ‘Elvetham’ stored under ambient conditions since 1880 were compared to a homonymous sample preserved in an on-farm situation in Denmark. DNA was isolated from single seeds and successfully applied to Amplified Fragment Length Polymorphism (AFLP) analysis of the accessions. Six primer pairs were used to determine the similarity between the two accessions based on 112 polymorphic bands. Furthermore, similarity among seven cultivars of fodder beets representing the main types used in Scandinavia at the end of the 19th century was determined. This analysis was based on 152 polymorphic bands. Differentiation among the seven cultivars was determined to a mean G _ST value of 0.438, while G _ST between the two ‘Elvetham’ accessions was 0.266. A principal coordinate analysis based on jaccards similarity index illustrates that the two ‘Elvetham’ accessions are different from each other. The differentiation is higher than the value found between two separate ‘Eckerndorfer’ accessions. The results indicate that the cultivated accession has changed. Additionally, the value of applying old dead seed material for documentation in gene banks is demonstrated. During the analysis it was found that DNA isolated from seeds and leaves behaved differently in the AFLP process, however, the two fractions assigned to their common accession.     

Genetic basis of species differentiation between <Emphasis Type="Italic">Coffea liberica</Emphasis> Hiern and <Emphasis Type="Italic">C. canephora</Emphasis> Pierre: Analysis of an interspecific cross

The phenotypic and genetic differentiation between the two related Coffea species (C. liberica Hiern and C. canephora Pierre) was examined. These species differed markedly in terms of leaf, inflorescence, fruit and seed characters. A genetic map of the interspecific cross Coffea liberica × C. canephora was constructed on the basis of 72 BC1 hybrids. Eighty-three AFLP markers, four inter simple sequence repeats (ISSR) and five microsatellites corresponding to Coffea liberica species-specific markers were mapped into 16 linkage groups. The total length of the map was 1502.5 cM, with an average of 16.3 cM between markers and an estimated genome coverage of 81%. The two species were evaluated relative to 16 quantitative traits and found to be significantly different for 15 of them. Eight QTLs were detected, associated with variations in petiole length, leaf area, number of flowers per inflorescence, fruit shape, fruit disc diameter, seed shape and seed length. Results on segregation distortion and the under-representation of particular markers were interpreted in terms of genome differentiation. The implications for the introgression of QTLs involved in advantageous morphological traits (number of flowers per inflorescence, fruit and seed shape) are discussed.     

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     

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.     

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.     

Diversity among landraces of Indian snapmelon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">momordica</Emphasis>)

Diversity among 36 snapmelon landraces, collected from 2 agro-ecological regions of India (9 agro-climatic sub-regions), was assayed using RAPD primers, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance and biochemical composition (TSS, ascorbic acid, titrable acidity). Typical differences among accessions were observed in plant and fruit characteristics and snapmelon germplasm with high titrable acidity and possessing resistance to downy mildew, Cucumber mosaic virus, Zucchini yellow mosaic virus, Papaya ringspot virus, Aphis gossypii and Meloidogyne incognita was noticed in the collection. RAPD based grouping analysis revealed that Indian snapmelon was rich in genetic variation and region and sub-region approach should be followed across India for acquisition of additional melon landraces. Accessions of var. agrestis and var. momordica clustered together and there was a separate cluster of the accessions of var. reticulatus. Comparative analysis of the genetic variability among Indian snapmelons and an array of previously characterized reference accessions of melon from Spain, Israel, Korea, Japan, Maldives, Iraq, Pakistan and India using SSRs showed that Indian snapmelon germplasm contained a high degree of unique genetic variability which was needed to be preserved to broaden the genetic base of melon germplasm available with the scientific community. N. P. S. Dhillon and Ranjana contributed equally to this work and are considered the first authors.     

<Emphasis Type="Italic">Oryza sativa</Emphasis> straw restricts <Emphasis Type="Italic">Phalaris minor</Emphasis> growth: allelochemicals or soil resource manipulation?

Unharvested stubbles or harvested straw of rice (Oryza sativa L.) gets incorporated into soil and interferes with the seedling growth of crop plants. In this paper, we investigated whether rice straw, either through releasing allelochemicals and/or through manipulating soil properties, influences seedling growth of Phalaris minor Retz., a non-native weed largely restricted to wheat (Triticum aestivum L.) fields. One hundred twenty grams of soil was amended with rice straw (0.5, 1, 2, 4, or 6 g/pot) and its effect on fresh shoot biomass of P. minor was examined. Any modification of rice straw phytotoxicity through the use of washed rice straw, activated charcoal, soil sterilization, or nitrogen fertilization was also studied. We carried out chemical and microbial analysis of soils to examine the role of soil properties in influencing P. minor growth. Incorporation of rice straw into soil suppressed the growth of P. minor through modifying soil properties. A dose-dependent increase in total phenolics was observed in soil amended with rice straw. Activated carbon or washing of rice straw, however, could not ameliorate the phytotoxic effects of rice straw. Our results provide initial evidence that rice straw restricts P. minor growth by manipulating soil chemical and microbiological properties. Authors contributions IJ conceived of and supervised the study, and wrote the paper; SK carried out the work.     

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.     

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.     

Ex situ collections of cultivated flax (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.) and other species of the genus <Emphasis Type="Italic">Linum</Emphasis> L.

An overview of ex situ collections of cultivated flax (Linum usitatissimum L.) and 53 other species of the genus Linum L. is presented. The names of 33 genebanks in 23 countries preserving Linum germplasm and the species preserved at these places are provided. World genebanks engage in ex situ preservation of about 48,000 accessions of cultivated flax (Linum usitatissimum). However, possibly only 10,000 accessions of these are unique. Of the ca. 200 species of the genus Linum, other than L. usitatissimum, 53 are reported to be represented in the world’s ex situ collections with a total of about 900 accessions. Of these, 279 accessions alone refer to the wild progenitor of cultivated flax, Linum bienne Mill. However, L. bienne from the region of origin of cultivated flax is rarely represented in genebank collections. References to publications describing the techniques for ex situ conservation and the infraspecific variation of cultivated flax are made. It is suggested that taxonomic species delimitation within the genus Linum is in need of clarification and that the variation in Linum species for ornamental use or use in breeding is investigated.     

Evaluation of genetic identity and variation of local apple cultivars (<Emphasis Type="Italic">Malus</Emphasis> × <Emphasis Type="Italic">domestica</Emphasis> Borkh.) from Spain using microsatellite markers

A collection of 114 local cultivars of Malus × domestica Borkh. from the Northwest of Spain and a group of 26 non-native commercial cultivars, were studied in order to determine the level of genetic differentiation between them, to know the population genetic structure of the three main production regions in North Spain, and to detect possible duplications, misidentifications, and intrusions of foreign cultivars into the present day Spanish apple gene pools. Ten primer pairs of microsatellite loci were selected from previous studies on apple. Allelic frequencies were used to estimate the average expected heterozygosity (Hexp) and F-statistics. A Principal Component Analysis and a Cluster Analysis were conducted in order to determine the relationships among cultivars. Considering all populations (Spanish and commercial cultivars) we found 119 alleles with the 10 microsatellites; 10 out of 119 were rare (with frequencies lower than 0.01). Spanish cultivars can be used as references for those alleles for further studies since they are available in Germplasm banks. Triploid cultivars were quite frequent, 39 out of 140, with the highest number occurring in Galicia (29 cultivars). North Spain presented a high variability for apple according to heterozygosity levels. Microsatellites have provided useful information about the singularity of Spanish apple cultivars, revealed six groups of synonymies and five introgressions of commercial cultivars.     

Analysis of Genetic Variation Among Accessions of <Emphasis Type="Italic">Pueraria montana</Emphasis> (Lour.) Merr. var. <Emphasis Type="Italic">lobata</Emphasis> and <Emphasis Type="Italic">Pueraria phaseoloides</Emphasis> (Roxb.) Benth. based on RAPD Markers

Pueraria montana var. lobata and P. phaseoloides originating from tropical Asia and parts of Oceania are ecologically and economically important legumes that are used as green manure, cover crop or forage plants. Conservation and use of plant genetic resources require an understanding of the extent and distribution of genetic diversity in any given region. In this study, genetic variation of five P. montana var. lobata and 16 P. phaseoloides accessions was analysed developing a random amplified polymorphic DNA (RAPD) marker methodology for Pueraria species and thereby creating basic data for follow-up research and the development of conservation strategies. Seeds were collected from native populations in Bac Kan Province, a mountainous region in Northeast Vietnam. P. montana var. lobata presented a high level of variation with 54.3% of the detected markers being polymorphic, whereas P. phaseoloides exhibited an intermediate to high level of variation (45.5%). The P. montana var. lobata accessions clustered in congruence with their eco-geographical origin. For P. phaseoloides no correspondence between sampling sites and genetic differentiation was found. Inter-population differentiation was measured as Jaccard's similarity coefficient (JSC). Mean JSC amounted to 0.35 in P. montana var. lobata and 0.52 in P. phaseoloides. Results are compared to other genetic studies of herbaceous legumes and conservation strategies are suggested.     

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.     

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.     

Community composition of ammonia-oxidizing bacteria in the rhizosphere of intercropped wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.), maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.), and faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

Cereal/cereal and cereal/legume intercropping systems are popular in the north, northwest, and southwest of China and often result in yield increases compared to monocropping. Rhizosphere interactions may play a significant role in the yield increases, particularly with respect to nutrient availability. The aim of this study was to investigate the effects of intercropping on N availability and community composition of ammonia-oxidizing bacteria in the rhizosphere of wheat, maize, and faba bean at different growth stages. Denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes was used to analyze the community composition of bacterial ammonia oxidizers belonging to β-proteobacteria. The results showed that intercropping with faba bean significantly increased nitrate concentrations in the rhizosphere of wheat and maize at the second sampling time (20 June) compared to monocropping or intercropping between maize and wheat. Intercropping significantly affected the community composition of ammonia-oxidizing bacteria in the rhizosphere compared to monocropping, and the effects were most pronounced in the maize/faba bean and wheat/maize intercropping systems when faba bean and wheat were at anthesis and maize was in seedling stage. In wheat/faba bean intercropping, the effects of intercropping on community composition of ammonia-oxidizing bacteria were less pronounced at the seedling stage of the two species but were significant at anthesis.     

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.