Characterization of European hazelnut (<Emphasis Type="Italic">Corylus avellana</Emphasis>) cultivars using SSR markers

World hazelnut production is based primarily on selections from the wild. In this study, we used 21 pairs of simple sequence repeat (SSR) primers to investigate genetic diversity in 270 clonal accessions of European hazelnut (Corylus avellana) representing a wide geographic range. Of the 270 accessions, 198 had unique fingerprints while 72 were duplicates. Based on the 198 unique accessions, the number of detected alleles per locus averaged 9.81 and observed heterozygosity (H_o) averaged 0.67. Of the 206 total alleles amplified, 20 were unique to a single accession. A genetic similarity matrix was constructed and the resulting dendrogram revealed four major geographical groups: Central European, Black Sea, English and Spanish-Italian. SSR alleles indicated the parentage of 31 accessions. The fingerprints are publicly available through the Germplasm Resources Information Network (GRIN) database. The identification of duplicate and mislabeled accessions will improve management of hazelnut genebanks, and information on genetic variation in hazelnut will assist the international research community. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Genetic diversity and differentiation in Chinese sour cherry <Emphasis Type="Italic">Prunus pseudocerasus</Emphasis> Lindl., and its implications for conservation

In this study, the genetic diversity and differentiation of 10 natural Prunus pseudocerasus Lindl. populations were investigated using inter-simple sequence repeat (ISSR) markers. Totally, 18 selected primers generated 150 loci, with an average of 8.33 bands per primer. The results showed that the percentage of polymorphic bands (PPB) was pretty low at the population level (PPB = 1.13–32%), but relatively high at the species level (PPB = 84%). Besides, a high level of genetic differentiation among populations was detected based on the gene differentiation coefficient (G _ST = 0.7118) and the hierarchical analysis of molecular variance (AMOVA) (Φ _ST = 64.53%, P < 0.001), in line with the low inter-population gene flow (N _m = 0.2025). Moreover, Mantel test revealed a significant correlation between genetic and geographic distances among the populations (r = 0.5272, P < 0.005). The high level of intraspecific genetic diversity was probably related with its life history traits, while its small population size and the resultant high levels of genetic drift and inbreeding might explain the low genetic diversity within populations. The relatively high inter-population genetic differentiation was largely attributed to its small population size, habitat fragmentation, the mode of pollen and seed dispersal, and geographic isolation. Based on the present study, conservation strategies were proposed to preserve this valuable natural germplasm resource.     

Genetic and bioclimatic variation in <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Solanum pimpinellifolium, due to its close relationship to S. lycopersicum, has been a genetic source for many commercially important tomato traits. It is a wild species found in the coastal areas of Peru and Ecuador. In this study, the genetic variation of S. pimpinellifolium was studied using the diversity found in 10 microsatellites in 248 plants spread throughout its entire distribution area, including Ecuador, which has been underrepresented in previous studies. Peruvian and Ecuadorian accessions are genetically quite differentiated. A possible cause of these differences could be the non-uniform nature of the coastal Ecuadorian and Peruvian climates, seeing as an important correlation between genetic differentiation and climate has been found. In addition, Ecuadorian and south Peruvian accessions have a lower genetic diversity and a higher homozygosity due to their higher autogamy, lower population size, and possible colonization bottlenecks. The Galápagos Islands population is an extreme case, with no diversity, likely caused by a recent colonization from the northern continental Ecuadorian region where genetically identical plants have been found. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Elena Zuriaga and José M. Blanca contributed equally to this work.     

Genetic Characterization of Brazilian Annual <Emphasis Type="Italic">Arachis</Emphasis> Species from Sections <Emphasis Type="Italic">Arachis</Emphasis> and <Emphasis Type="Italic">Heteranthae</Emphasis> using RAPD Markers

The genus Arachis is divided into nine taxonomic sections. Section Arachis is composed of annual and perennial species, while section Heteranthae has only annual species. The objective of this study was to investigate the genetic relationships among 15 Brazilian annual accessions from Arachis and Heteranthae using RAPD markers. Twenty-seven primers were tested, of which nine produced unique fingerprintings for all the accessions studied. A total of 88 polymorphic fragments were scored and the number of fragments per primer varied from 6 to 17 with a mean of 9.8. Two specific markers were identified for species with 2n = 18 chromosomes. The phenogram derived from the RAPD data corroborated the morphological classification. The bootstrap analysis divided the genotypes into two significant clusters. The first cluster contained all the section Arachis species, and the accessions within it were grouped based upon the presence or absence of the ‘A’ pair and the number of chromosomes. The second cluster grouped all accessions belonging to section Heteranthae.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

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.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Anthropogenic and natural causes influencing population genetic structure of <Emphasis Type="Italic">Juniperus procera</Emphasis> Hochst. ex Endl. in the Ethiopian highlands

Juniperus procera is economically highly important but threatened tree species. It is the only species among 67 taxa in the genus Juniperus that naturally grows in Africa and south of the equator extending up to 18°S in Zimbabwe. Ethiopia is assumed to host the largest J. procera populations, which are also believed to have high genetic variation owing to their wide ecological amplitude. This study assessed genetic variation at AFLPs of J. procera populations in the Ethiopian highlands. In the study six populations, namely Chilimo, Goba, Menagesha-Suba, Wef-Washa, Yabelo and Ziquala were included. A total of 20–24 trees from each population were investigated based on 128 AFLP band positions. AMOVA revealed that most of the variation (94%) resided within populations of J. procera suggesting extensive gene flow among populations which is attributable to the outcrossing mating system and effective gene transport mechanisms of the species. However, genetic differentiation among populations was still significant (P < 0.05), and the differentiation was significantly (P < 0.05) correlated with geographic distance. All population pairs were significantly (P < 0.05) differentiated except for Menagesha-Suba and Wef-Washa. These two populations also showed the highest gene diversity (H _j = 0.301 and H _j = 0.297, respectively). These results are in accordance with historical records that claim the establishment of the Menagesha-Suba juniper population as plantation of seedlings from Wef-Washa back in fifteenth century.     

<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Isozyme polymorphism and genetic differentiation in natural populations of an endemic tetraploid species <Emphasis Type="Italic">Avena maroccana</Emphasis> in Morocco

A wild tetraploid oat Avena maroccana Gdgr. was collected from the 11 populations in the periphery of Rommani and Casablanca geographic groups of Morocco. Genetic diversity of the species was investigated using six allozyme systems. Allelic frequencies were scored representing eight polymorphic and five monomorphic loci. Coefficient of gene differentiation (Gst) was 0.3019, which indicated great genetic differentiation. The number of alleles per locus was 2.6154, the percentage of polymorphic loci was 61.54, and the expected heterozygosity was 0.2462 in all populations. Genetic diversity in A. maroccana was high in comparison to self-pollinated species. In total, nine heterozygotes resulting from outcrossing were found in the progeny from M1, M3, M4, M22 and M26. The population of M7 had peculiar alleles Pgd–2SS and Pgd-1SS in high frequency. M9 had the lowest level of diversity out of the 11 populations. Geographic and genetic distances between all the populations were not significantly correlated with each other (r = 0.0996). Cluster analysis showed that two groups, (M1, M22, M2 and M4) and (M3, M23, M8, M5 and M26) were apparently differentiated. Two populations of the Casablanca group, M7 and M9 were independent from each other, and were separated distinctly from the other populations. Genetic diversity of the Rommani and Casablanca groups was almost the same in all the parameters. This was due to the similar man-made habitat such as roadside or rich fertile soil and brown clay soils. The population size of A. maroccana was small and restricted to the narrow central Morocco with great genetic differentiation so that genetic diversity may be reflected from the results of genetic drift and outcrossing heterozygote segregation.     

Genetic diversity of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) assessed with individuals and bulks on agarose gels

About 65 years ago, more than 150 Swiss maize landraces (Zea mays L. ssp. mays) of the flint type were collected and conserved ex situ. Due to the climatically and culturally diverse environment of the Alps, a considerable genetic diversity of this material was assumed. To prove this, an efficient method was required to carry out genetic profiling of all the accessions in the Swiss Gene Bank. Simple sequence repeat marker (SSR) profiling in combination with the visualization of the polymerase chain reaction (PCR) products on agarose gels was chosen. Here a set of 19 different landrace accessions was analyzed to: (i) investigate their genetic diversity, (ii) investigate and display the population structure and (iii) determine whether DNA bulks rather than single plants can be used for such analyses. Four repeated samples of one accession were found to be much closer to one another than to the rest of accessions. Furthermore, specific alleles were identified for several accessions. The PCR products of the bulked DNA samples represented only a small part of the variation revealed by the analysis of individuals. Loci with four base repeat motifs performed better in the analysis of bulks than loci with other repeat motifs. The correlation between genetic distance matrices, based on the analysis of individuals and bulks, respectively, was significant. Thus, the single plant approach allowed for sufficient differentiation of accessions, and DNA bulks visualized on agarose gels led to correlated genetic distances although a limited number of alleles were detected. Although the limited resolution of agarose gels likely causes some bias, profiling of larger sets with the individual plant approach appears feasible and more informative compared to the bulk analysis we conducted.     

<Emphasis Type="Italic">Ziziphus </Emphasis><Emphasis Type="Italic">spina-christi</Emphasis> (L.) Willd.: a multipurpose fruit tree

Neglected and underutilized species often play a vital role in securing food and livestock feed, income generation and energy needs of rural populations. In spite of their great potential little attention has been given to these species. This increases the possibility of genetic erosion which would further restrict the survival strategies of people in rural areas. Ziziphus spina-christi is a plant species that has edible fruits and a number of other beneficial applications that include the use of leaves as fodder, branches for fencing, wood as fuel, for construction and furniture making, and the utilization of different parts e.g. Fruits, leaves, roots and bark in folk medicine. Moreover, the plant is adapted to dry and hot climates which make it suitable for cultivation in an environment characterized by increasing degradation of land and water resources. Lack of research in Z. spina-christi hinders its successful improvement and promotion. Therefore, studies are needed to fully exploit this species. This article aims at summarizing information on different aspects of Z. spina-christi to stimulate interest in this crop which is of importance in Sudan and other countries of the semi-arid tropics.

Karyotypic studies in wild germplasm of <Emphasis Type="Italic">Arachis</Emphasis> (Leguminosae)

The karyomorphology for eight diploid species of Arachis belonging to three sections has been described for the first time, Sect. Extranervosae: A. macedoi (2n = 20m) and A. retusa (2n = 14m + 6sm); Sect. Heteranthae: A. sylvestris (2n = 16m + 4sm); Sect. Procumbentes: A. chiquitana (2n = 18m + 2sm); Sect. Arachis: A. cruziana (2n = 18m + 2sm), A. herzogii (2n = 18m + 2sm), A. simpsonii (2n = 20m) and A. williamsii (2n = 20m). A pair of satellited chromosomes was observed in all species. A chromosomes were found in A. chiquitana, A. herzogii and A. simpsonii. Karyotypic differences between sections were observed, but not enough to establish a characteristic karyotype pattern for each section. However, the species may be differentiated by the presence of A chromosomes, the type and position of satellites, and the karyotype formulae. These results are discussed with regard to karyotype evolution in Arachis to contribute to understanding the role of chromosome changes in the evolution of the genus.     

A study of genetic diversity of sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) in Vietnam and Cambodia estimated by RAPD markers

Sesame (Sesamum indicum L.) is a traditional oil crop cultivated throughout South East Asia. To estimate the genetic diversity of this crop in parts at the region, 22 sesame accessions collected in Vietnam and Cambodia were analyzed using 10 RAPD markers. The 10 primers generated 107 amplification products of which 88 were polymorphic fragments (83%). Genetic diversity of all populations was H_t = 0.34 when estimated by Nei’s genetic diversity and species diversity was H′_sp = 0.513 when estimated by Shannon diversity index. Genetic distance ranged from 0.03 to 0.43, with a mean genetic distance of 0.23. The unweighted pair group method with arithmetic averages (UPGMA) cluster analysis for the 22 accessions divided the material in four groups. The dendrogram revealed a clear division among the sesame accessions based on their geographical region. Interestingly, some geographically distant accessions clustered in the same group, which might indicate the human factor involved in the spreading of sesame varieties. The high level of polymorphism shown suggests that RAPD techniques can also be useful for the selection of parents in sesame (Sesamum indicum L.) breeding program and for cultivar differentiation.     

Resistance gene analog polymorphisms (RGAPs) in wild emmer wheat (<Emphasis Type="Italic">Triticum dicoccoides</Emphasis>) and their ecological associations

Using the 8 specific primer pairs based on the conserved motifs of plant resistance genes, the plant disease resistance gene analog polymorphisms (RGAPs) in 15 wild emmer wheat (Triticum dicoccoides) populations from Israel had been detected. High genetic variations at the RGAP loci were observed in T. dicoccoides populations. A total of 254 discernible bands were obtained among 115 accessions, and 192 bands (75.6%) were polymorphic. Each genotype had a unique banding profile, and the genetic similarity coefficient ranged from 0.094 to 0.862. In T. dicoccoides, the proportion of polymorphic loci (P), the genetic diversity (He) and Shannon’s information index were 0.756, 0.362 and 0.541, respectively. The proportion of polymorphic loci (P) per population averaged 0.732 (range: 0.515–0.932); genetic diversity (He) averaged 0.271 (range: 0.212–0.338); and Shannon’s information index averaged 0.404 (range: 0.310–0.493). The coefficients of genetic distance (D) among populations averaged 0.107 (range: 0.043–0.178), and the results of Mantel test (r = 0.168, P = 0.091) showed that the estimates of genetic distance were geographically independent. Neighbor-joining cluster analysis suggested that the genetic relationships of T. dicoccoides populations were associated with their ecogeographic distribution. The hierarchical analysis of molecular variance (AMOVA) and the coefficient of gene differentiation (G _ST ) values revealed that most of the variations were presented within populations, although significant differences among populations and regions were also detected. The values of P and Shannon’s information index were negatively correlated with the two factors: Tdd (day–night temperature difference) and Ev (mean annual evaporation), whereas they were positively correlated with one water factor: Rn (mean annual rainfall). The correlation matrix between He in the RGAPs and geographic variables contained 20 significant (P < 0.05) correlations. The present study established that T. dicoccoides in Israel had a considerable amount of genetic variations at RGAP loci at least partly correlated with ecological factors.     

Genetic variation in the endangered Rutaceae species <Emphasis Type="Italic">Citrus hongheensis</Emphasis> based on ISSR fingerprinting

Citrus hongheensis is a critically endangered species endemic to the Honghe river region in southeastern Yunnan, China. Its genetic diversity and differentiation were investigated using Inter-Simple Sequence Repeat (ISSR) markers. One hundred primers were screened, and a total of 245 loci were amplified from seven natural populations by 13 informative and reliable primers. Of these 245 ISSR loci, 233 were polymorphic and the detected variations revealed a relatively high level of intraspecific genetic diversity. At the population level, the mean percentage of polymorphic loci (PPB) was 36.50%, while the average expected heterozygosity (He) and Shannon diversity index (Ho) were 0.1327 and 0.1972, respectively. At the species level (across all populations), PPB was 95.10%, while He and Ho were 0.3520 and 0.5195, respectively. A high Gst value (0.6247) indicated that there is significant differentiation among populations, which was confirmed by AMOVA analysis (Φst = 0.6420). Pairwise genetic identity (I) values among populations ranged from 0.6341 to 0.7675, with a mean of 0.7008. We propose that the high level of genetic differentiation may be the result of habitat fragmentation and limited gene flow (Nm = 0.1502). For effective in situ conservation and population restoration of C. hongheensis it will be important to maintain historical processes, including high outbreeding rates, sufficient gene flow, and large effective population sizes.     

Analysis of population substructure,genetic differentiation and phylogenetic relationships among selected Asiatic <Emphasis Type="Italic">Vigna</Emphasis> Savi species

Random amplified polymorphic DNA markers were used to study sub-structure and genetic differentiation amongst 31 populations (seven cultivated and 24 wild populations) belonging to 14 Asiatic Vigna species. Ten pre-selected RAPD primers generated 152 polymorphic amplification products. Estimates of polymorphism indices were higher for the wild taxa in comparison to the cultivated forms. F_ST values between populations ranged from 0.111 to 0.801 and Nei’s genetic diversity values between and within species varied from 0.26 to 0.70 and 0.04 to 0.56 respectively. The high F_ST and F_CT values indicated strong subdivision of populations and high differentiation among species. Analysis of molecular variance was performed by grouping the populations conforming to specific species. AMOVA was also performed separately to better resolve the differentiation of species within mungo–radiata complex. Molecular phylogenetic relationships amongst the species of radiata–mungo complex; namely, black gram (V. mungo (L.) Hepper), green gram (V. radiata (L.) Wilczek), V. radiata var. sublobata, V. radiata var. setulosa, V. mungo var. silvestris and V. hainiana, were studied through cluster analyses. Two distinct groups were recognized within the complex, with population samples of V. hainiana forming one cluster. Further, V. hainiana appeared to be equidistant to both V. radiata and V. mungo.     

Effects of long-term mineral fertilization on microbial biomass,microbial activity,and the presence of <Emphasis Type="Italic">r</Emphasis>- and <Emphasis Type="Italic">K</Emphasis>-strategists in soil

Long-term effects of mineral fertilization on microbial biomass C (MBC), basal respiration (R _B), substrate-induced respiration (R _S), β-glucosidase activity, and the r–K-growth strategy of soil microflora were investigated using a field trial on grassland established in 1969. The experimental plots were fertilized at three rates of mineral N (0, 80, and 160 kg ha⁻¹ year⁻¹) with 32 kg P ha⁻¹ year⁻¹ and 100 kg K ha⁻¹ year⁻¹. No fertilizer was applied on the control plots (C). The application of a mineral fertilizer led to lower values of the MBC and R _B, probably as a result of fast mineralization of available substrate after an input of the mineral fertilizer. The application of mineral N decreased the content of C extracted by 0.5 M K₂SO₄ (C _ex). A positive correlation was found between pH and the proportion of active microflora (R _S/MBC). The specific growth rate (μ) of soil heterotrophs was higher in the fertilized than in unfertilized soils, suggesting the stimulation of r-strategists, probably as the result of the presence of available P and rhizodepositions. The cessation of fertilization with 320 kg N ha⁻¹ year⁻¹ (NF) in 1989 also stimulated r-strategists compared to C soil, probably as the result of the higher content of available P in the NF soil than in the C soil.     

Exploitation of <Emphasis Type="Italic">Malus</Emphasis> EST-SSRs and the utility in evaluation of genetic diversity in <Emphasis Type="Italic">Malus</Emphasis> and <Emphasis Type="Italic">Pyrus</Emphasis>

A total of 8117 suitable SSR-contaning ESTs were acquired by screening from a Malus EST database, among which dinudeotide SSRs were the most abundant repeat motif, within which, CT/TC followed by AG/GA were predominant. Based on the suitable sequences, we developed 147 SSR primer pairs, of which 94 pairs gave amplifications within the expected size range while 65 pairs were found to be polymorphic after a preliminary test. Eighteen primer pairs selected randomly were further used to assess genetic relationship among 20 Malus species or cultivars. As a result, these primers displayed high level of polymorphism with a mean of 6.94 alleles per locus and UPGMA cluster analysis grouped twenty Malus accessions into five groups at the similarity level of 0.6800 that were largely congruent to the traditional taxonomy. Subsequently, all of the 94 primer pairs were tested on four accessions of Pyrus to evaluate the transferability of the markers, and 40 of 72 functional SSRs produced polymorphic amplicons from which 8 SSR loci selected randomly were employed to analyze genetic diversity and relationship among a collection of Pyrus. The 8 primer pairs produced expected bands with the similar size in apples with an average of 7.375 alleles per locus. The observed heterozygosity of different loci ranged from 0.29 (MES₉₆) to 0.83 (MES₁₃₈), with a mean of 0.55 which is lower than 0.63 reported in genome-derived SSR marker analysis in Pyrus. The UPGMA dendrogram was similar to the previous results obtained by using RAPD and AFLP markers. Our results showed that these EST-SSR markers displayed reliable amplification and considerable polymorphism in both Malus and Pyrus, and will contribute to the knowledge of genetic study of Malus and genetically closed genera.     

Use and variation of <Emphasis Type="Italic">Pandanus tectorius</Emphasis> Parkinson (<Emphasis Type="Italic">P. fascicularis</Emphasis> Lam.) along the coastline of Orissa,India

Pandanus tectorius Parkinson ( P. fascicularis Lam.) of the family Pandanaceae constitutes one of the major bioresources of Ganjam coast, Orissa; used mainly in small scale perfume industry for aromatic compound extracted from the male inflorescences. In order to establish genetic diversity, if any related to perfume yield, samples of P. tectorius representing male populations from seven locations representing populations I–VII along the coastline of Orissa, India, were analysed for somatic chromosome number, 4C genomic DNA content, randomly amplified polymorphic DNA (RAPD) as well as phytochemicals. The somatic chromosome number in all the populations I–VII was 2n = 60. The chromosomes were of minute size without showing any remarkable structural variation. Like wise the average 4C DNA content was 5.09 pg (≅4,912 Mbp) that showed no intra- or inter-population differences. Out of 54 decamer primers tested, a total of 1,260 amplicons were obtained from 34 primers accounting 43.49% polymorphism. Molecular phylogenetic analysis of the seven populations revealed two distinct branches, with populations II and III in one and the rest populations in the other branch of the phylogenetic tree. It was important to note that the unique populations II and III confined to the Ganjam coast of Orissa having RAPD markers: OPA 09–940 bp, OPA 09–705 bp, OPC 14–1,500 bp, OPC 14–700 bp, OPC 20–1,475 bp, OPC 20–1,350 bp, OPC 20–920 bp and OPC 20–700 bp, were distinguished form the rest of the populations. The aforesaid populations (II and III) are well known to produce aroma of high quality and yield, composed of primarily phenyl ethyl methyl ether (66.8–83%) and terpinen-4-ol (5–12%) along with a number of other phyto-chemical compounds that support the flourishing perfume industry and livelihood of the local people in the region. The findings underscored the possible role of local eco-geography in contributing to the micro-evolution of unique high perfume yielding genotypes of P. tectorius that represented populations II and III at Ganjam coast, which were genetically distinct from the rest of the populations revealed by RAPD analysis.