The insertion of the Agrobacterium rhizogenes rolC gene in tomato (Solanum lycopersicum L.) affects plant architecture and endogenous auxin and abscisic acid levels

In the present paper we report on the effects of the insertion of the Agrobacterium rhizogenes rolC gene in the tomato (Solanum lycopersicum L., formerly Lycopersicon esculentum Mill.) cultivar Tondino. Several transgenic lines were successfully obtained, between which two clones, rolC1 and rolC3, were chosen for the analysis of morpho-productive traits as well as of the endogenous levels of auxin and abscisic acid. Consistent with the known phenotypic effect of this gene, the transformed tomato plants were significantly shorter than the corresponding controls. On the other hand, even if yield was not affected by the transformation in terms of average number of fruits produced, fruit weight was significantly lower in the transgenics with respect to the controls. Therefore, insertion of the rolC gene does not lead to an improvement in plant productivity.     

The risks of success in quality vegetable markets: Possible genetic erosion in Marmande tomatoes (Solanum lycopersicum L.) and consumer dissatisfaction

Following consumer complaints about the quality of modern varieties of tomato, landraces have strengthened their quality markets. In Spain, two tomato landraces, ‘Montserrat’ and ‘Pera Girona’, are grown in contiguous areas and have different market niches. We used amplified fragment length polymorphism (AFLP) to characterize 13 accessions of Montserrat, 14 accessions of Pera Girona, and 4 control varieties. We found a narrow genetic base for ‘Montserrat’ and ‘Pera Girona’ (8.5% of polymorphic loci) and no differences between the landraces. We studied agronomical and sensory traits to determine why the two landraces continue to have separate market niches. We found high variability among accessions within each landrace and overlapping among accessions of both landraces for all traits except fruit morphology. Consumers probably came to associate the organoleptic quality of these landraces with their external traits, but due to spontaneous crossings and introgressions these relations have been lost. Selection within landraces will be necessary to reestablish the link between morphology and sensory value and to consolidate these quality markets.     

Identification of Italian landraces of bean (Phaseolus vulgaris L.) using an image analysis system

Colour, shape and size of whole seeds and their spots of some Italian landraces of bean (Phaseolus vulgaris L.) were measured using a specifically developed macro, based on image analysis library KS-400 V 3.0 (Carl Zeiss, Germany).     

Genetic diversity in local Tunisian pears (Pyrus communis L.) studied with SSR markers

Few records are available about local Tunisian pear cultivars characterized by low chilling requirements and adaptation to dry conditions. In this work, seven SSRs derived from apple were successfully transferred to 25 local Tunisian pear genotypes and 6 common varieties of Pyrus communis cultivated in Europe. The 7 SSRs used amplified a total of 36 fragments. All the microsatellites except one seem to amplify more than one locus in some of the genotypes studied. Only 12 different fingerprinting patterns could be distinguished among the 25 Tunisian cultivars studied indicating a high number of synonymies. The mean expected and observed heterozygosities in the 25 Tunisian cultivars analyzed averaged 0.71 indicating a high level of genetic diversity among the local Tunisian pear germplasm. These markers will be useful to optimize the conservation of this highly threatened germplasm.     

Translocation of heavy metals to tomato (Solanum lycopersicom L.) fruit irrigated with treated wastewater

A field experiment was conducted to investigate the extent of translocation of heavy metals to tomato (Solanum lycopersicom L. cvs. “GS₁₂” and “RS₅₈₉₉₅₆”) fruit produced in an open field near to Abu-Nusiar Wastewater Treatment Plant, Amman-Jordan. Seedlings were planted during the seasons of 1999 and 2000 and furrow irrigated with different mixtures of potable water to treated wastewater {100%:0% (1:0, control); 25%:75% (1:3); 50%:50% (1:1); 0%:100% (0:1)}. Tomato fruit, soil and water were examine for heavy metals concentrations, and changes in the pH and electrical conductivity (EC) of the soil were tested. The obtained results revealed an increase in the concentrations of Cu, Mn and Fe, in the soil correlated with high concentrations found in the wastewater. Additionally, an increase in the pH and EC in the soil was observed with increasing the proportions of wastewater. Results of tomato fruit analysis showed an increased concentration of Fe, Cu, Ni, Mn and Zn in the cultivar “RS₅₈₉₉₅₆”, whereas, an increased concentration of Mn and Zn were detected in the cultivar “GS₁₂”, but no accumulation of Cd and Pb in both cultivars. The accumulation of heavy metals in fruit was below the Jordanian standard limits, thus, the use of treated wastewater in irrigation might be feasible.     

Evaluation of ‘Tonda di Giffoni’ hazelnut (Corylus avellana L.) clones

‘Tonda di Giffoni’ is among the most highly appreciated Italian hazelnut (Corylus avellana L.) cultivars. Due to its round kernels and excellent processing quality, it was awarded a Protected Geographical Indication (PGI) from the European Union. To identify clones expressing improved nut and production qualities, a ‘Tonda di Giffoni’ clonal selection programme was conducted across hazelnut orchards in the Irno valley of Italy from 1995 to 2006. One hundred different clones were selected and propagated in a replicated trial under similar climate, soil, and cultural conditions. From this work, the 29 best clones were identified and from 2006 to 2008 their agronomic and pomological characteristics were observed. Microsatellite or simple sequence repeat (SSR) markers were used to successfully confirm true-to-type identity of the clones. Traits evaluated included flowering time (anthesis), bud break, suckering, trunk diameter, nut and kernel characteristics and productivity (yield). Best linear unbiased predictions for clone means and estimates of intraclass correlation coefficient were obtained using R environment, lme4 and ggplot2 packages. Five clones superior to that of the standard of ‘Tonda di Giffoni’ were identified in this study. Furthermore, yield and number of suckers produced showed sufficient variability to likely be exploited for breeding. The selected clones express features useful for both growers and the processing industry and will be propagated and planted in hazelnut orchards for further study and commercial production.     

Phenotypic variability and genetic structure in plum (Prunus domestica L.), cherry plum (P. cerasifera Ehrh.) and sloe (P. spinosa L.)

In the present study, phenotypic variability of 80 plum (Prunus domestica L.) varieties maintained in the French National Plum Collection was evaluated with 19 quantitative traits. In addition, genetic diversity and genetic structure was studied in three plum species (P. domestica L., Prunus cerasifera Ehrh. and Prunus spinosa L.) using chloroplast DNA (cpDNA) markers and five single sequence repeat (SSR) loci. Based on phenotypic traits, some varieties, such as mirabelle plums, grouped together. Bayesian structure analysis was used to identify different genetic groups, whereby damson plums were clearly distinguished from greengage plums. When examining the three species together, a higher level of cpDNA allelic richness was found in P. cerasifera and in P. spinosa than in P. domestica where only five cpDNA haplotypes were detected in the national plum collection, with one main haplotype that accounted for 80% of the varieties studied. P. domestica cpDNA haplotypes tended to group together with P. cerasifera haplotypes whereas most of P. spinosa haplotypes formed a separate cluster. SSR markers were somewhat able to distinguish the three species. These results provide some clues as to the origin of plum and the various plum varieties. Our results also provide useful information for the management of plum genetic resources.     

Putting back what we take out,but how much?: Phosphorus and nitrogen additions to farmed Leucadendron ‘Safari Sunset’ and Leucospermum ‘Succession’ (Proteaceae)

Proteaceae are adapted to low-nutrient soils in the various regions where they occur. However, harvesting of flowering stems for the cut-flower industry must eventually cause soil nutrient depletion sufficient to reduce yields. Different N forms, and N and P concentrations were supplied to two Proteaceae cultivars (Leucadendron ‘Safari Sunset’ and Leucospermum ‘Succession’) in a controlled fertigation experiment, and appropriate concentrations for maximum growth with minimum nutrient accumulation or loss were determined. Small additions of N (0.025–0.1 mM) significantly improved growth of both cultivars growing on Strandveld sandy soil. Larger additions of N (up to 2 mM N) resulted in poor growth (both cultivars) and N accumulation in the soil (Safari Sunset). Small additions of P (<10 μM) significantly improved growth of both cultivars and resulted in no accumulation or loss of P in the soil. Larger additions of P (up to 500 μM) resulted in poor growth, P toxicity symptoms and P leaching from the upper soil layers. Best N forms in descending order of both plant visual appearance and vegetative yield were: urea ≥ ammonium nitrate > ammonium sulphate > calcium nitrate. Phosphorus toxicity symptoms were associated with increased concentrations of leaf P, Ca and Fe. Under conditions of maximum growth (10 μM P and 0.1 mM N) Safari Sunset removed 18 ± 0.6 g N, 1.5 ± 0.1 g P, 5.3 ± 0.6 g K and Succession removed 5.5 ± 0.2 g N, 0.3 ± 0.02 g P, 3.1 ± 0.5 g K over 6 months. At maximum growth, plants acquired more N and P amounts than were supplied, but supplying higher N and P concentrations adversely affected growth. Thus, a more complex or slow-release form of N and P than urea and soluble phosphate, respectively, may provide enough N and P to replace losses from the farm soil at the low concentrations required for proteas.     

RAPD analysis of genetic diversity among and within Portuguese landraces of common white bean (Phaseolus vulgaris L.)

Seventeen landraces of common beans (Phaseolus vulgaris L.) sampled in the North and Centre of Portugal were analyzed at population level for 689 RAPD loci amplified by using forty random primers. The two different parameters used to estimate the genetic variability in and between samples indicate that the inter-population component of the genetic variability is mainly responsible for the diversity found, since only about a 10% would be of an intra-population nature. In addition, the gametic disequilibrium was estimated and reached an average value of 40% for the different combinations of pairs in the 689 loci studied taking the 17 samples as a whole population. Self-pollination, genetic drift and adaptation would thus be favouring the formation of multilocus associations. In addition, the fingerprinting study suggests that each landrace produced unique amplification products allowing it to be distinguished from the other tested genotypes, but, nevertheless, the landraces show a considerable genetic similarity (56% and 70.5% using two different methods). The Neighbour-Joining dendrogram did not show any relation between the geographical distribution of landraces and genetic distance. The results suggest that these Portuguese landraces conserved an important genetic diversity that can be useful to widen the genetic base of currently cultivated beans.     

Genetic variation in tomato populations from four breeding programs revealed by single nucleotide polymorphism and simple sequence repeat markers

Genetic variability in modern crops is limited due to domestication and breeding. To investigate genetic variation in different populations, 216 tomato (Solanum lycopersicum L.) cultivars, hybrids, and elite breeding lines from four breeding programs were genotyped using single nucleotide polymorphism and simple sequence repeat markers. Of 47 markers analyzed, 72.3% were polymorphic in the whole collection of 216 genotypes and 51.06–59.57% showed polymorphisms in individual populations. However, genetic variation was narrow in all four populations. Nei's genetic distance varied from 0.0422 to 0.1135 between populations and from 0.0085 to 0.3187 between lines in individual populations. Cluster and principal coordinate analysis indicated that the four populations could be grouped into three clades. Lines from Shenyang Agricultural University and China Agricultural University population formed the first clade, lines from Beijing Vegetable Research Center were in the second clade, and lines from Nunhems were in the third clade. This was further supported by population structure analysis using STRUCTURE2.2, and suggested that a lack of germplasm exchange might exist among breeding programs. It might be the reason that the progress of developing new varieties with significant improvement of horticultural traits in China is slow in recent years.     

Chlorocholine chloride application effects on photosynthetic capacity and photoassimilates partitioning in potato (Solanum tuberosum L.)

Treatment of potato (Solanum tuberosum L.) with chlorocholine chloride (CCC) applied twice as a foliar spray 25 and 30 days after planting has shown to decrease shoot and stolon growth but increase tuber yield. However, the regulatory role of CCC on translocation of recently fixed photoassimilates into different parts of potato plants has not been fully illustrated. In this study, ¹⁴C-isotope labelling technique was used to estimate the photosynthetic capacity and photoassimilate partitioning among leaves, stems, roots + stolons, and tubers of potted potatoes treated with 1.5 g l⁻¹ CCC. CCC treatment significantly increased tuber dry mass but reduced leaf dry mass. CCC-treated leaves had significantly higher chlorophyll and carotenoid contents and assimilated 22.0% more ¹⁴CO₂ per leaf dry mass than the controls. Compared with the control, CCC treatment reduced the translocation of ¹⁴C-photoassimilates into leaves, stems and roots + stolons but increased that into tubers. CCC-treated leaves exported 14.6% more ¹⁴C-photoassimilates into other parts of the plants. In addition, CCC treatment reduced ¹⁴C-soluble sugar and ¹⁴C-starch accumulation in leaves and stems but enhanced them in tubers and roots + stolons. Collectively, the results indicate that CCC treatment significantly improves the photosynthetic capacity of potato leaves and promotes photoassimilates partitioning into tubers thereby enhancing tuber growth.     

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.     

Application of chlorophyll fluorescence for the diagnosis of salt stress in tomato “Solanum lycopersicum (variety Rio Grande)”

To study the response of tomato (Solanum lycopersicum cv. Rio Grande) to salinity, the effect on plant growth, water relations, stomatal conductance and Chlorophyll fluorescence was investigated. Tomato plants were grown in peat culture under controlled conditions and submitted during 28 days to saline stress ranging from 0 to 25, 50, 100, 150 and 200 mM of NaCl. At the end of the experiment period, plant growth was significantly decreased with increasing salinity.     

Genetic diversity of bitter gourd (Momordica charantia L.) genotypes revealed by RAPD markers and agronomic traits

Bitter gourd or bitter melon (Momordica charantia L.) is considered as minor cucurbitaceous vegetable in spite of having considerable nutritional and medicinal properties. Although some reports on genetic diversity based on morphological characterization are available, no work has been conducted to estimate genetic diversity using molecular markers in this crop. In the present study, 38 genotypes of M. charantia including few commercially cultivars collected from different parts of India based on agro-ecological zones were analysed for diversity study both at morphological and molecular levels. Genomic DNA was extracted from young healthy leaves following the procedure of Doyle and Doyle [Doyle, J.J., Doyle, J.L., 1990. A rapid DNA isolation procedure from small quantity of fresh leaf material. Phytochem. Bull. 119, 11–15]. Pair-wise comparison of genotypes was calculated as per the procedure of Jaccard [Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223–270]. Dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA) and the computation for multivariate analysis was done using the computer programme NTSYS-pc Version 2.0 [Rohlf, F.J., 1998. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.01. Exeter Software, Setauket, NY, USA]. Diversity based on yield related traits and molecular analysis was not in consonance with ecological distribution. Among 116 random decamer primers screened 29 were polymorphic and informative enough to analyse these genotypes. A total of 208 markers generated of which 76 (36.50%) were polymorphic and the number of bands per primer was 7.17 out of them 2.62 were polymorphic. Pair-wise genetic distance (GD) based on molecular analysis ranged from 0.07 to 0.50 suggesting a wide genetic base for the genotypes. The clustering pattern based on yield related traits and molecular variation was different.     

Genetic diversity and population's structure in Tunisian strawberry tree (Arbutus unedo L.)

A growing interest in the use of the Strawberry tree (Arbutus unedo L., Ericaceae) has been recently reported for industrial, pharmaceutical and chemical fields. However, the bulk material comes from natural populations because of the lack of selection of interesting cultivars. In Tunisia, A. unedo populations are severely destroyed due to deforestation and over-collecting. The species occurs in small scattered populations decreasing progressively in size. Yet, no conservation or improvement programs are attempted to preserve and promote the potential value of this resource. In this work, we assessed the genetic diversity of nine Tunisian populations of A. unedo L. from different bioclimates, using 65 polymorphic RAPD loci. The analysis of the genetic variation within and among populations is primordial to elaborate conservation and improvement programs. A low genetic diversity within a population estimated by both Nei's (He) and Shannon's diversity (H′) indices (0.155 < He < 0.248; 0.229 < H′ < 0.364) was observed due to genetic drift and selfing. At the species level, the amount of the within population variation estimated by Shannon's index (H_POP/H_SP = 0.686) and the molecular variance (80.67%) was higher than that among populations. A moderate genetic differentiation among populations (Φ_ST = 0.193 and G_ST = 0.314) which could be attributed to the long seed distance dispersal was detected. The UPGMA dendrogram based on Φ_ST values showed three clusters each including populations without relationship to bioclimatic or geographical origin indicating that differentiation occurs at a local space scale. The in situ protection measures should be made appropriately according to a population within bioclimates. The ex situ conservation and the selection of genotypes should involve extensive collection of seeds or cuttings from the within populations rather than among them.     

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’.     

Agrobacterium × plant factors influencing transformation of ‘Joseph's coat’ (Amaranthus tricolor L.)

A protocol is developed for Agrobacterium-mediated genetic transformation of Amaranthus tricolor via explant co-cultivation with Agrobacterium rhizogenes. Bacteria-plant specific factors which influenced transformation were optimized. Of the two Agrobacterium strains employed, LBA9402 was more infectious compared to A4. Bacterial suspensions grown overnight with 100 μM acetosyringone and experiencing O.D.₆₆₀ = 0.6 followed by dilution to a density of 10⁹ cells ml⁻¹ were the most effective. Explants from garden-grown plants were more responsive than those from in vitro cultures; stem internodes being better than leaves. Immersion of the pre-pricked explants in bacterial suspension resulted in a markedly higher transformation frequency compared to the direct injection method. The infection of internode explants with the LBA9402 strain followed by co-cultivation on growth regulator-free MS medium (MS0) for 5 days resulted in emergence of hairy roots up to a maximum frequency of 97.22%. Roots were individually cultured in MS0, but fortified with bactericidal antibiotic (500 μg ml⁻¹ cefotaxime). Rhizoclones showing prolific growth were renewed through successive subcultures in MS0. Opine gene expression was revealed by positive agropine and mannopine synthesis in all selected transformed rhizoclones. Shoot regeneration from root clones, capable of auxin-independent growth and opine proficiency, was stimulated in MS augmented with 2.0 mg l⁻¹ zeatin. pRi TL–DNA rolB and pRi TR–DNA man2 ORF were detected in leaf tissues of regenerated plants from selected hairy root clones through PCR amplification. The implication of such findings is discussed on the possibility of conferring protection to crop amaranths against biotic stress challenges, particularly due to insects, viruses or fungal pathogens.     

Impact of iron stress on biomass,yield, metabolism and quality of potato (Solanum tuberosum L.)

In order to study the influence of variable iron on biomass, economic yield and role of iron in potato (Solanum tuberosum) cv. Chandramukhi metabolism, plants were grown in refined sand at variable iron ranging from 0.001 to 2.0 mM. Exposure of potato plants to Fe stress (i.e. a Fe concentration different from 0.1 mM) shows retarded growth, decreased chlorophyll concentration and Hill reaction activity, induced changes in enzyme activities and concentration of Fe and Mn. The visible symptoms of iron deficiency appeared on day 15 at 0.001 mM Fe as chlorosis of young leaves. The excess of iron (at >0.1 mM Fe) appeared later, after 25 days and the chlorosis was observed on older leaves. Both deficiency (0.001 mM) and excess (>0.1 mM) of iron reduced the tuber yield, deteriorating its quality by lowering the concentration of sugars, starch and protein nitrogen and increasing the accumulation of non-protein nitrogen and phenols in tubers.     

Effect of distance from early flowering pollinizers ‘Maypole’ and ‘Dolgo’ on ‘Fiji’ fruit set

An apple orchard consisting of a single cultivar under the condition of natural pollination must have suitable pollinizers such as Crab apples to ensure stable fruit production. We selected ‘Maypole’ and ‘Dolgo’ as pollinizers for the cultivar ‘Fuji’, and investigated the rate of fruit and seeds in ‘Fuji’ fruits produced by pollen of the pollinizers. We developed a method for tracing pollen flow based on the leaf color of progeny and S-RNase allele of ‘Maypole’, and on Simple Sequence Repeat (SSR) analyses of ‘Maypole’ and ‘Dolgo’. These were powerful tools for determining the distance insects (mainly Osmia cornifrons) carry pollen from the pollinizers to ‘Fuji’. Although the fruit set of ‘Fuji’ apples was not reduced with increasing distance between ‘Fuji’ apple trees and pollinizers (probably due to pollen flow from other commercial cultivars planted outside the area), the rate of ‘Fuji’ apple fruit produced by the pollen of the pollinizers decreased with increasing distance. The rate of fruit produced by the pollinizers was 84% and 77% when ‘Fuji’ was 2.5 m from ‘Maypole’ and ‘Dolgo’, respectively, and 71% and 64% when ‘Fuji’ was 5 m from ‘Maypole’ and ‘Dolgo’, respectively, but was reduced to 47% and 39% when ‘Fuji’ was 10 m from ‘Maypole’ and ‘Dolgo’, respectively. However, the spacing could cause reduced fruit size, and require extra fruit thinning for producing large fruits. It could also cause reduction of the yield for increasing the planting area of pollinizers. We recommend that pollinizers should be planted not more than 10 m from ‘Fuji’.