Root restriction-induced limitation to photosynthesis in tomato (Lycopersicon esculentum Mill.) leaves

Root restriction often depresses photosynthetic capacity and the mechanism for this reduction, however, remains unclear. To identify the mechanism by which root restriction affects the photosynthetic characteristics, tomato (Lycopersicon esculentum Mill.) seedlings were subjected to root restriction stress with or without supplemental aeration to the nutrient solution. With the development of the root restriction stress, CO₂ assimilation rate was decreased only in confined plants without supplemental aeration. There were also significant decreases in leaf water potential, stomatal conductance (g_s), intercellular CO₂ concentration (C_i), and increases in the stomatal limitation (l) and the xylem sap ABA concentration. Meanwhile, the maximum carboxylation rate of Rubisco (V_cmax) and the capacity for ribulose-1,5-bisphosphate regeneration (J_max) also decreased, followed by substantial reductions in the quantum yield of PSII electron transport (Φ_PSII). Additionally, root restriction resulted in accumulation of carbohydrates in various plant tissues irrespective of aeration conditions. It is likely that root restriction-induced depression of photosynthesis was mimicked by water stress.     

Assessment of genetic diversity among mango (Mangifera indica L.) genotypes using RAPD markers

Knowledge about the extent of genetic diversity/relatedness in mango germplasm is vital for developing coherent strategies for future gains in productivity. The genetic diversity/relatedness among mango cultivars/genotypes developed in Pakistan has not been investigated previously. We have assessed the genetic diversity among 25 mango genotypes/cultivars using randomly amplified polymorphic DNA (RAPD). Sixty random ten-mer primers were surveyed, out of which 45 yielded amplicons in all the genotypes. Genetic similarity between genotypes/cultivars was in the range of 64–89% with an average of 74%. Similarly, the genetic relatedness among all variants derived from a mango cultivar Chaunsa was in the range of 81.18–88.63%. These coefficients were utilized to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA). The genotypes were grouped into three (A, B, C) clusters. Generally, genotypes originating from Pakistan were grouped in cluster ‘A’ while cluster ‘B’ primarily composed of southern India as well as Florida cultivars. Kensington Pride was the most distantly related genotype which grouped with Maya and Yakta, forming a distinct cluster ‘C’.     

Role of surface morphology in determining the ripening behaviour of tomato (Lycopersicon esculentum Mill.) fruits

Surface characteristics of fruit mainly determine the diffusibility of gases across its boundary and also the extent to which moisture being lost from the fruit. The present study has been carried out with an objective to find out the association between the ripening related processes and surface morphology of tomato (Lycopersicon esculentum Mill.) fruit. Tomato fruits of two contrasting varieties; Pusa Gaurav (a slow ripening type) and Pusa Ruby (a relatively fast ripening type) were evaluated for physiological loss in weight (PLW), rate of respiration, effect of hypoxia on ripening, dimension of stem scar region and distribution of lenticels along with hair base cell of trichomes on the surface of the fruits. Results showed that fruits of Pusa Gaurav showed significantly lower PLW, rate of respiration, surface area of stem scar region, density of lenticels along with hair base cell of trichomes and slower ripening under hypoxia condition in comparison to Pusa Ruby. More resistance towards gaseous exchange and therefore lower O₂ to CO₂ ratio inside the fruits of Pusa Gaurav in contrast to Pusa Ruby in view of the differences in the surface morphological features in these two varieties could possibly be assigned as one of the reasons responsible for the obtained varietal difference in ripening and related changes.     

Effects of NaCl application to hydroponic nutrient solution on fruit characteristics of tomato (Lycopersicon esculentum Mill.)

NaCl was applied to nutrient solution (5 dS m⁻¹ versus 1.4 dS m⁻¹ in the control) of hydroponically-grown tomato and its effects on taste grading and chemical composition of fruit were investigated. Taste panels indicated NaCl treatment increased sweetness, acidity, umami (i.e. the taste of deliciousness) and overall preference. Hexose concentration of the fruit grown on NaCl treated plants significantly increased; and at the same time, chloric ion, organic and amino acids in general had higher concentrations in NaCl treated plants than the control. Our results showed that (1) consumer grading of the tomato fruit was influenced not only by sugar content but also by the organic and amino acids; (2) increased concentration of soluble solids in the fruit of NaCl treated plants was not the result of simple overall condensation due to the reduction of water transport. The relation of diversified consumer preference, fruit chemical composition, and appropriate evaluation of tomato fruit are also discussed.     

Resistance of 57 greenhouse-grown accessions of Lycopersicon esculentum and three cultivars to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

The aim of this study was to evaluate resistance to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) by antixenosis on 57 Lycopersicon esculentum Mill. accessions from the Horticultural Germplasm Bank (HGB) of Universidade Federal de Viçosa and by the three commercial cultivars (Santa Clara, Moneymaker and TOM-601) under greenhouse conditions. A randomized complete block design was used with three replications. Infestations with T. absoluta adults were performed weekly and the following characteristics were evaluated: number of small, large and total mines/leaf and % of leaves mined at days 60, 75 and 90 after planting. Low infestation occurred at days 60 and 75 but at day 90, infestation was sufficient to evaluate insect damage. Based on these data it was concluded that only accessions HGB-674 and HGB-1497 appeared to be the most promising. In addition, to determine possible chemical causes of resistance, hexane extracts were analyzed at day 90 by gas chromatography/mass spectrometry and the major peaks identified by a mass spectral database using similarity index. Nine hydrocarbons, viz., hexadecane, heptadecane, eicosane, tricosane, 2-methyltricosane, tetracosane, hexacosane, octacosane and triacontane were identified in the hexane extracts in many samples. Tricosane, tetracosane and hexacosane presented significant correlations with the leaves mined. Only tricosane presented a negative correlation with the number of small mines (r = −0.28), total number of mines (r = −0.27) and % of leaves mined (r = −0.22). However, tetracosane and hexacosane presented significant positive correlations (r = 0.25 and 0.24, respectively) with the % of leaves mined.     

Assessment of genetic diversity of Latvian and Swedish sweet cherry (Prunus avium L.) genetic resources collections by using SSR (microsatellite) markers

Three previously described highly polymorphic SSR (microsatellite) primer pairs were tested on 126 sweet cherry (Prunus avium L.) accessions to adapt a fast, reliable method for preliminary screening of sweet cherry germplasm collections and to compare two sweet cherry germplasm collections: at the Latvia State Institute of Fruit-Growing, Dobele (LIFG-Dobele) and at the Division of Horticultural Genetics and Plant Breeding at Balsgård, Department of Crop Sciences, Swedish University of Agricultural Sciences (SLU-Balsgård). The SSR loci were highly polymorphic with 4–10 different alleles and 5–18 genotypes. Heterozygosity values ranged from 0.431 to 0.809, gene diversity (PIC) values ranged from 0.400 to 0.753, and the discriminating power of each locus varied from 0.631 to 0.894. The combined discriminating power of all loci was highly effective (0.996). Sixteen identical accession groups with the same allele profile were discovered in both collections. This study demonstrated that SSR fingerprinting with the three primer pairs tested, can be used for preliminary characterization of sweet cherry germplasm collections.     

Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It is native to Iran and spread from Iran to other areas. In this study amplified fragment length polymorphism (AFLP) was used to detect intra- and inter-population genetic diversity of pomegranate. A group of 67 accessions belonged to 4 populations from Iran was studied using eight primer combinations. A total of 221 scorable bands were amplified, of which, 118 (54.13%) were polymorphic. Resolving power (Rp) ranged from 5.70 to 9.21, and the average of polymorphism information content (PIC) per primer pair was 0.40. According to Nei's gene diversity and allelic statistics, Isfahan population had a highest genetic diversity (H = 0.3646, I = 0.5327, N_e = 1.6467). Coefficient of gene differentiation between populations (GST) was 0.124, indicated that mainly proportion of genetic variation (87.6%), was within populations and the remaining (12.4%) of the variation was among populations that, also supported by analysis of molecular variance (AMOVA). The gene flow (Nm) varied from 0.969 to 10.404 between pair-wise populations and was 3.504 among all of the populations. The Jaccard similarity coefficient between individuals ranged from 0.26 to 0.88. The UPGMA dendrogram clustered all 67 accessions into 6 groups. In some cases accessions from same region were grouped together but in most cases, there was gene exchange. To study the genetic relationships among populations, a principal coordinate analysis (PCoA) based on Nei's genetic distances was performed. Results of this study showed that AFLP marker can be a useful tool for investigating the genetic diversity of pomegranate genotypes.     

Rose (Rosa hybrida L.) EST-derived microsatellite markers and their transferability to strawberry (Fragaria spp.)

The ‘Genome database for Rosaceae (GDR)’ provides a large collection of expressed sequence tags (ESTs) harboring simple sequence repeats (SSRs) from several Rosaceae genera, including Rosa (rose). Primer pairs flanking SSR were designed for 312 unique Rosa ESTs based on GDR database. Eight rose (Rosa hybrida L.) genotypes were tested for PCR amplification, and 287 (92%) of the primer pairs generated allele-specific PCR bands that were readily scored. From 183 (63.7%) primer pairs that evidenced polymorphic alleles among the eight rose cultivars, 20 pairs evidencing EST sequence homology to known gene functions and high levels of polymorphism were selected and utilized for DNA fingerprinting and genetic diversity assessments of 47 rose hybrids. A total of 202 polymorphic bands were scored and generated unique fingerprints for each rose hybrid. The Nei–Li genetic similarity coefficients among 1081 pair-wise comparisons of 47 cultivars exhibited a broad range of genetic variations from 0.30 (‘Grand King’ and ‘Carnival’) to 0.99 (‘First Red’ and ‘Red Champ’). UPGMA cluster analysis divided 47 hybrids into five major groups and two sub-groups. The cross-species transferability of 273 EST-SSR primer pairs was evaluated using four genotypes of the strawberry, a genus member of the Rosaceae family. PCRs on the DNA samples of strawberry were successful for 165 primer pairs; among these, 123 pairs amplified 243 polymorphic bands. As surrogates of the marker transfer, the phenetic relationship among the four strawberry genotypes was evaluated. Genetic similarity coefficients varied from 0.78 (‘Maehyang’ and ‘Janghyee’) to 0.64 (‘Janghyee’ and ‘Pragana’). The results of cluster analysis showed that the three octaploid strawberry cultivars were quite similar, whereas the diploid ‘Pragana’ was related distantly at the genomic DNA level. The EST-SSR markers developed in the present study can be efficiently utilized for genetic diversity studies in Rosaceae.     

Assessment of genetic diversity and species relationships in eggplant (Solanum melongena L.) using STMS markers

Ninety six accessions including 92 of Solanum melongena and four related non-tuberous species (Solanum insanum, S. incanum, S. integrifolium and S. sysimbriifolium) were taken for the assessment of genetic diversity using 23 STMS primers. Eleven of the 23 primers tested showed polymorphism. The number of alleles per primer ranged from three to six with an average of 4.4. S. melongena had maximum average similarity with its closely related species, S. insanum (0.67) and minimum average similarity with the wild species, S. sysimbriifolium (0.50). The two weedy species S. incanum and S. integrifolium showed more average similarity value of 0.62 and 0.61, respectively with the cultivated S. melongena. S. insanum. S. incanum and S. integrifolium were relatively similar to each other with similarity index value of 0.61 (between S. insanum and S. incanum), 0.63 (between S. insanum and S. integrifolium) and 0.62 (between S. incanum and S. integrifolium). In contrast S. sysimbriifolium was most divergent with the similarity value of 0.49, 0.47 and 0.51 with S. insanum, S. incanum and S. integrifolium, respectively. The closely related species S. insanum and S. incanum, which clustered along with S. melongena accessions, being crossable with cultivated species, constitute important sources of genes that can be introgressed by backcross breeding. Molecular markers can be employed to identify the hybrids and also to monitor introgression of the useful genes.     

Assessment of genetic diversity in common bean (Phaseolus vulgaris L.) germplasm using amplified fragment length polymorphism (AFLP)

AFLP technique was applied to assess genetic diversity among 44 common bean accessions that included 6 exotic accessions, 15 Indian land races and 23 released varieties. Eight AFLP primer pairs were used that produced 820 products of which 698 were polymorphic (85.12%). Wide variations were observed among all the accessions for the number of amplification products, percent polymorphism and average polymorphism information content (PIC). The Jaccard's similarity indices (J) based on the AFLP profiles were subjected to UPGMA cluster analysis. The dendrogram generated revealed seven major groups. Seventeen out of 23 released varieties were restricted to clusters VI and VII. The value of r = 0.934 in Mantel's test for cophenetic corrlelation applied to the cluster analysis indicated the high fitness of the accessions to a group. The germplasm used in the present study had narrow genetic base, although moderate to high genetic diversity was observed. The details of diversity analysis and the potential use of Indian common bean accessions in common bean breeding programme are provided in the present study.     

Analysis of genetic diversity among some Persian walnut genotypes (Juglans regia L.) using morphological traits and SSRs markers

There is a high diversity among cultivated walnut trees in Iran due to its long time of seed propagation and vast area of cultivation. In this study some morphological characters as well as Simple Sequence Repeat (SSRs) markers were used to analyze the genetic diversity and relationships among 31 Iranian walnut genotypes along with four foreign cultivars. The nut weight ranged from 7.52 to 17.73 g, kernel weight from 4.00 to 9.83 g, and kernel percentage ratio from 38.78 to 67.05% among studied genotypes. In SSRs analysis, nine primer pairs were tested that produced 39 alleles ranging from 2 to 8, with a mean value of 5.10 allele per primer. The Iranian genotypes showed relatively high diversity both for their SSRs loci and morphological traits. Although the foreign cultivars (‘Serr’, ‘Vina’, ‘Franquette’ and ‘Lara’) clustered with each other, they also laid close and within the Iranian genotype. The results of the study provided us with valuable diversity among our genotypes which could be used for breeding studies and also showed the power of genetic markers for analysis and evaluation of this diversity.     

Confirmation of hybrid origin in Arisaema (Araceae) using molecular markers

A population of hybrids between Arisaema triphyllum subsp. stewardsonii and A. dracontium was investigated using molecular markers to document the hybrid origin. Total genomic DNA was extracted from A. triphyllum, A. dracontium, and the hybrids, and subjected to sequence analysis of various regions of intergenic spacer and genes of chloroplast or intergenic spacer and genes of nuclear ribosome for single nucleotide polymorphisms (SNPs). Clustering was performed using the unweighted pair group method with averages (UPGMA) tested by bootstrap (BS). Hybrid origin could not be easily confirmed in some regions. However, dendrograms constructed with a combined sequence analysis of A. triphyllum 26S ribosomal RNA gene [sequence identification (SI) 467] plus A. tortuosum phytochrome C-like (phyC) gene (SI 468) were very similar to dendrograms constructed from sequences of all regions. This suggests that selecting SI 467 and SI 468 would be practical to identify hybrid origins involving two parental species. Clustering of hybrids together with the female parent in most target regions suggests that, in Arisaema, cpDNA is considered maternally inherited.     

Fingerprinting of three typical macrosperma Italian lentil (Lens culinaris Medik.) landraces using fluorescence-based AFLP markers

Italian lentil landraces are principally cultivated for self or local consumption. Most of them are disappearing, particularly macrosperma types by being less required by the market. A pre-requisite for the conservation and the efficient use of genetic resources is the better understanding of the extent and the distribution of the existing genetic variation, useful for future breeding programmes. Our study was undertaken to analyse and quantify the genetic diversity within and among three macrosperma Italian lentil landraces (Onano, Altamura and Villalba), using fluorescent AFLP markers. AFLP markers generated information to differentiate among closely related genotypes and group within the same cluster individuals belonging to the same landrace. The total genetic diversity (H_T), the genetic diversity within population (H_S) and the extent of differentiation between populations (D_ST) were 0.198, 0.155 and 0.043, respectively. The fixation index (G_ST = 0.219) showed that about 78% of the observed total genetic variation can be attributed to within population differences and around 22% is due to differences among populations. The gene flow estimate (N_m = 1.774) and the mean genetic distance value (0.077) suggested narrow genetic base among the analysed populations, confirming the tendency of Italian lentil landraces to group together. The present study showed that fluorescence-based AFLP technique is a biotechnological tool that can provide significant insights for research in genetic diversity of lentil landraces and their subsequent conservation and utilization in breeding programs.     

AFLP-based genetic relationships in the Mediterranean myrtle (Myrtus communis L.)

Myrtle is an important plant species of the Mediterranean maquis, and is widely exploited for its aromatic properties. It is used in Italy for the production of a typical liqueur, for cut foliage and as an ornamental pot plant. We report the use of amplified fragment length polymorphism (AFLP) profiling to estimate genetic similarities within myrtle germplasm collected from six Italian regions, and from the Botanical Gardens of six other Mediterranean countries (including the outgroup Myrtus communis subsp. tarentina). Five AFLP primer combinations identified 122 polymorphic fragments analysing 92 individual samples, most of them (56%) were informative in discriminating among the populations. The AFLP patterns indicated that the majority of the variation occurs among rather than within populations (G_ST = 0.61). A neighbour-joining (NJ) tree separated the populations into two main branches: the first one grouped some of the Italian populations with those from Spain and Portugal; the second one included a Southern Italian subcluster together with samples from Greece, Israel, France and Croatia. A principal coordinate analysis supported the two major branches identified in the NJ analysis and showed the separation of Western and Eastern Mediterranean populations along the first axis. The Italian populations did not cluster in a single clade, but rather form distinct regional groups. The present analysis suggests that Italy represents a botanical transition zone between the Western and Eastern Mediterranean region in Myrtus communis.     

Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop improvement strategies

A genetic analysis of 38 diverse Indian bitter gourd (Momordicacharantia var. charantia, and var. muricata) accessions was performed using 29 RAPD and 15 ISSR markers. RAPD primers yielded 208 amplicons of which 76 (36.5%) were polymorphic providing an average of 2.6 amplicons per primer. RAPD amplicons per primer ranged from 3 (OPE-19, OPW-09) to 15 (OPW-05), and varied in size from 200 bp to 3000 bp. Fifteen ISSR primers provided a total of 125 bands of which 94 (74.7%) were polymorphic. Polymorphic ISSR markers ranged from 0 (UBC-841) to 12 (UBC-890) providing a mean of 6.3 amplicons per primer that ranged in size from 150 bp to 2700 bp. Nevertheless, the concordance among bitter gourd accession groupings after cluster analysis was relatively high (r = 0.77), indicating that RAPD- and ISSR-based diversity assessments in this germplasm array were generally consistent. The M.charantia var. charantia (domesticated) and var. muricata (wild, free-living) accessions examined were genetically distinct, and these differences provided for the development of strategies for genetic analyses and crop improvement in this species.     

Effect of root applied 28-homobrassinolide on the performance of Lycopersicon esculentum

The roots of 20 days old seedlings of tomato (Lycopersicon esculentum Mill.) at the time of their transplantation, were dipped in 10⁻⁸, 10⁻⁷, 10⁻⁶ or 10⁻⁵ M of 28-homobrassinolide (HBR) for 15, 30 or 45 min and were allowed to grow in earthen pots, in a net house. The leaves of the plants, at days 30 and 60, possessed elevated quantities of nitrate reductase (NR), carbonic anhydrase (CA) and the contents of chlorophyll. The values for all the above characteristics were significantly higher than that of the water-fed control. The fruits borned at the treated plants were more in number and possessed a lower quantity of ascorbic acid than the control. Moreover, the fruits at ripening, had higher levels of lycopene and β-carotene. Among the treatments, 15 min feeding of 10⁻⁸ M HBR proved best.     

Microsatellite markers based assessment of genetic diversity in Iranian olive (Olea europaea L.) collections

To study the genetic variation in Iranian olive collections and some foreign olive cultivars, 47 accessions of 18 local cultivars from 6 olive collections of Iran (Roudbar, Zanjan, Ahvaz, Dezful, Kazeroon and Shiraz), were analyzed along with 30 imported cultivars using 16 microsatellite primer pairs. All the used microsatellite loci revealed polymorphism in the studied genotypes, except GAPU14 and GAPU113 markers. Fourteen microsatellite primers amplified 126 polymorphic alleles in the 87 selected olive accessions. The average number of alleles per locus was 9, ranging from 3 to 14. Polymorphic information content (PIC) was 0.85. The genetic similarity based on Jaccard coefficient ranged from 0.15 to 1. The genetic relationships among accessions were investigated using cluster analysis and principal component analysis (PCA). Most of the accessions with the same name were grouped together; some exceptions were also observed. As expected, close relationship was observed among accessions within same cultivar. Most of the Iranian olive accessions were clustered to a main distinct group. Two-dimensional scatter plot of principal component analysis revealed a clear separation of most of the Iranian olives from Syrian and other introduced cultivars. These suggest that Iranian cultivars have different origin related to West Mediterranean basin cultivars and have evolved independently from the others. Between and within Iranian and foreign cultivars (cultivars including three or more accessions) genetic diversity was analyzed using analysis of molecular variance (AMOVA). AMOVA revealed higher within cultivar genetic variation (62.76%) as compare to that between cultivar variations (37.24%). The intra- and inter-cultivar variance tested by permutation test showed significant genetic variation at both levels. The high level within cultivar genetic variance could be due to mislabeling and presence of homonyms in cultivars produced by vegetative propagation from original plants.     

RAPD markers reveal polymorphism among some Iranian pomegranate (Punica granatum L.) genotypes

In this study RAPD markers were used to determine the diversity level among 24 Iranian pomegranate genotypes. One hundred decamer random primers were used for PCR reactions, among which 16 showed reliable polymorphic patterns. These primers produced 178 bands, of which 102 were polymorphic. Cluster analysis of the genotypes was performed based on data from polymorphic RAPD bands, using Jaccard's similarity coefficient and UPGMA clustering method. The highest and lowest similarities detected between genotypes were 0.89 and 0.29, respectively. At a similarity of 60%, the genotypes were divided into four sub-clusters. Cophenetic correlation coefficient between similarity matrix and cophenetic matrix of dendrogram was relatively high (r = 0.9) showing the goodness of fit of the dendrogram. RAPD markers showed to be a useful tool for studying the genetic diversity of pomegranate.     

Ameliorative effect of polyamines on the high temperature inhibition of in vitro pollen germination in tomato (Lycopersicon esculentum Mill.)

Effect of polyamines on in vitro pollen germination at high temperatures in tomato (Lycopersicon esculentum Mill.) was investigated. Pollen germination and tube growth were significantly inhibited at 33°C and 35°C compared to those at 25°C. This inhibition was reversed by the addition of spermidine or spermine in the germination medium. Spermidine at 0.5 mM was slightly more effective than spermine at 0.05 mM. Spermidine at 0.05 and 0.5 mM and spermine at 0.05 mM slightly increased pollen germination rates at 25°C. Spermidine at 5 mM and spermine at 0.5 M were inhibitory to pollen germination, regardless of incubation temperatures. Spermidine also promoted germination of pollen grains incubated at 38°C for 1 and 3 h and then at 25°C for the rest of the 20 h incubation period. The effect was higher at 0.5 mM than at 0.05 mM. Treatment of spermidine to intact flowers 1 day before anthesis was also effective in ameliorating the high temperature inhibition of in vitro pollen germination on the polyamine-free medium. Here, the optimum concentration was 5 mM. These results demonstrate that polyamines can counteract the inhibitory effects of high temperature on pollen germination. They also suggest that the endogenous level of polyamines in germinating pollen grains is an important factor for the pollen germinability at high temperature.