Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Efficiency of SNP and SSR-based analysis of genetic diversity,population structure,and relationships among cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.) germplasm from East Africa and IITA inbred lines

The extent of genetic diversity and relatedness of cowpea germplasm from East Africa are poorly understood. A set of 13 microsatellites (SSR) and 151 single nucleotide polymorphisms (SNPs) markers were applied to assess the levels of genetic diversity in a sample of 95 accessions of local cowpea germplasm and inbred lines of Vigna unguiculata. The average genetic diversity (D), as quantified by the expected heterozygosity, was higher for SSR loci (0.52) than for SNPs (0.34). The polymorphic information content was 0.48 for SSR and 0.28 for SNP while the fixation index was 0.095 for SSR and 0.15 for SNPs showing moderate differentiation and high gene flow among cowpea accessions from East African countries. The results of data analysis of both SSR and SNP markers showed similar clustering patterns suggesting a substantial degree of association between origin and genotype. Principal coordinate analysis (PCoA) with SSR and SNP markers showed that accessions were grouped into two and three broad groups across the first two axes, respectively. Our study found that SNP markers were more effective than SSR in determining the genetic relationship among East African local cowpea accessions and IITA inbred lines. Based on this analysis, five local cowpea accessions Tvu-13490, Tvu-6378, Tvu-13448, Tvu-16073, and 2305675 were identified to be tightly clustered sharing several common alleles with the drought tolerant variety Danila when analyzed with SSR and SNP markers. The findings will assist and contribute to future genetic diversity studies aimed at the genetic improvement of local Eastern Africa cowpea accessions for improved overall agronomic performance in general and breeding for drought tolerant in particular.     

Seasonal Changes of Shoot Carbohydrates and Growth Characteristics of ‘Trakya İlkeren’ Grape Variety (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

This research was conducted to determine the changes in the seasonal carbohydrates of annual shoots and growth characteristics of 10 years old ‘Trakya ?lkeren’ grape variety. Grapevines are grafted on 5BB and 5C rootstocks and they were grown in heavy clay soil conditions. In the study, changes of sugar, starch and total carbohydrates were determined from bud burst to dormancy. While sugar, starch and total carbohydrates showed significant differences (P < 0.01) there were no differences between rootstocks. Although sugar, starch and total carbohydrates of shoots decreased from bud burst to blooming, they increased thereafter until vegetative growth stopped. Total carbohydrates of annual shoots have been found to accumulate in period from blooming to harvest. Total and mean leaf area, shoot length and diameter, total chlorophyll and chlorophyll a/b ratio showed significantly variation among rootstocks and phenological stages (P < 0.01). Whereas the highest chlorophyll contents were found in the blooming, it was found at the lowest through to harvest. Overall mean and total leaf area, shoot length and shoot diameter, total chlorophyll content and chlorophyll a/b ratio were significantly higher on 5BB grafted vines. In the study, 5BB rootstocks are found suitable in terms of shoot carbohydrate accumulation and growth characteristics for ‘Trakya ?lkeren’ grape variety.     

Effects of Iron Sulfate,Zinc Sulfate,Iron Chelate,Powder Sulphur and Humic Acid Applications on Vegetative Growth of Sweet Cherry (<Emphasis Type="Italic">Prunus avium</Emphasis> L.)

This study was carried out in ?scehisar district of Afyonkarahisar on ‘0900-Ziraat’ sweet cherry cultivar for two successive years in 2011 and 2012. Ten different applications consisting of 80?g FeSO₄.7H₂O tree^?1, 20?g FeEDDHA tree^?1, 420?mL TK?-Hümas tree^?1, 80?g FeSO₄.7H₂O + 420?mL TK?-Hümas tree^?1, 200?g powder sulphur (S) tree^?1, 80?g FeSO₄.7H₂O + 200?g powder S tree^?1, 25?g ZnSO₄.7H₂O tree^?1, 25?g ZnSO₄.7H₂O + 420?mL TK?-Hümas tree^?1 and 25?g ZnSO₄.7H₂O + 200?g powder S tree^?1 were subjected for determination of their effect on vegetative growth of sweet cherry. In this study, shoot diameter, shoot length, leaf total chlorophyll content, leaf area and leaf iron (Fe) and zinc (Zn) contents were obtained. Shoot length, shoot diameter and leaf area showed general increases in 80?g FeSO₄.7H₂O + 200?g powder S tree^?1 and 25?g ZnSO₄.7H₂O + 200?g powder S tree^?1 treatments.The results clearly indicated that he treatments had different effects on the shoot length and shoot diameter although there was a differences between the experimental years. Particularly, significantly positive effects of the treatments on leaf total chlorophyll was apparent for the second year’s observations. In addition, Fe and Zn concentrations in leaf were markedly increased in response to the treatments.     

Dietary apigenin suppresses IgE and inflammatory cytokines production in C57BL/6N mice

Flavonoids ubiquitously exist in plants, vegetables, fruits, and teas. We evaluated the effect of dietary apigenin, one of the well-known flavonoids, on the immune system in C57BL/6N mice. Mice were fed experimental diets containing apigenin for 2 weeks. After the experimental period, there was no significant difference in body and organ weights between the control and the apigenin group. The total immunoglobulin (Ig) E levels in mice fed apigenin were significantly suppressed, whereas levels of IgG, IgM, and IgA were not affected. We also examined the effect of the apigenin diet on cytokine expression in mice sera using a cytokine array. The production of regulated upon activation normal T cell expressed and secreted (RANTES) and soluble tumor necrosis factor receptor I (sTNFRI) in mice sera was down-regulated by the apigenin diet. These results suggest that a diet containing apigenin can reduce serum IgE and inflammatory cytokines such as RANTES and sTNFRI in mice.     

Maintaining Genetic Resources of Peach Palm (<Emphasis Type="Italic">Bactris gasipaes</Emphasis> Kunth): The Role of Seed Migration and Swidden-fallow Management in Northeastern Peru

Knowledge of the effects of farmer practices on population genetic parameters of peach palm (Bactris gasipaes Kunth) is relevant to the improvement and conservation of the palm’s genetic resources. Microsatellite markers were used to assess genetic diversity and population structure of peach palm in swidden-fallow agroforestry systems in northeastern Peru. The study covered eight communities, comprising two study areas 160 km apart – one occupied by indigenous Amerindians and the other by mixed race campesinos. Simultaneous analysis of an ex situ peach palm germplasm collection provided a means to compare population genetic parameters. Farmers who were surveyed on seed selection practices for peach palm reported that an average of only four palms (4.3 for campesino and 1.5 for indigenous populations) were used to provide seed for the establishment of the forest gardens sampled. As expected, inbreeding coefficients observed within communities were relatively high (f = 0.105 − 0.210), however, observed heterozygosities within communities were also high (0.625–0.741). A metapopulation approach was used to describe migration within and among regions, implying a hierarchical structure of gene flow which maintains relatively high levels of genetic diversity. Seed migration was found to occur over longer distances (≤600 km) and at a higher frequency (46% of palms sampled) in the indigenous study area, and a proportionally greater number of alleles was found (49 vs. 43 over three loci) with twice as many private alleles occurring only in the indigenous populations. The farmers’ practice of preserving remnant palms through successive swidden generations may have contributed to the maintenance of alleles by reducing the severity of founder effects. Although the campesino study area exhibited a significant (20% of the variation; p < 0.01) isolation-by-distance relationship across 35 km distance, in general, both study populations had relatively limited genetic structure (θ = 0.012–0.03), which is believed to have resulted from the exchange of seeds over long distances and periods of time.     

Sedimentary characterization of Tagus estuarine beaches (Portugal)

Background, Aim and Scope  The distribution of sediments in estuarine beaches is controlled by the interactions between sediment supply, hydrodynamic processes and human intervention. The main purpose of this study is to characterize the sediments of Tagus estuarine beaches in order to understand their origin and to contribute to a better knowledge of the Tagus estuary sediment budget. Methods  Surface sediment samples were collected across beach profiles and sand grain size analysis was performed by dry sieving. Grain size statistics for the median (d₅₀) and standard deviation (SDM) were obtained using the Moment method. This study was complemented by a qualitative evaluation of the sediment composition. Cross-shore topographic surveys were conducted for selected sampling sites. Results  Tagus estuarine beach sediments are mainly composed of quartz sand particles which are fine-grained and well sorted near the mouth of the estuary and medium to coarse-grained and moderately sorted in the inner domain. Compositional results show evidence of active anthropogenic sediment sources, especially in the coarser fractions. Discussion  The analysis of the textural and compositional characteristics of beach sediments in the inner estuarine domain is compatible with local sedimentary sources, while a marine signature is present at the mouth and inlet channel sediments. In the inner domain, differences in the sedimentary processes are represented by the textural characteristics of the sediments, such as the sorting degree and the gravel content. Sediment characteristics also reflect human intervention in the system, with the introduction of anthropogenic and allochthonous particles and the mixture of sediments from different sources. Conclusions  The sediments of the inner Tagus estuarine beaches are derived from local Plio-Pleistocene outcrops while inlet and outer estuary beaches reveal a dominant marine source. Beach textural variability observed in the inner domain is not related to wave forcing gradients, but mainly to variations in the sedimentary processes along the estuarine margins and to human intervention. Results show that the Tagus estuarine beaches depended, almost exclusively, on sediment input from local sources until the last century. With increasing human occupation, sediment transfers became dominated by anthropogenically related activities mainly connected with the occupation of estuarine margins and dredging. Recommendations and Perspectives  Further studies should extend the present level of knowledge in what concerns sand transport patterns through additional compositional and geochemical analysis, and the development of new techniques in order to allow the quantitative evaluation of the impact of human activities on the sediment budget.     

Soil compaction and forest floor removal reduced microbial biomass and enzyme activities in a boreal aspen forest soil

Soil enzymes are linked to microbial functions and nutrient cycling in forest ecosystems and are considered sensitive to soil disturbances. We investigated the effects of severe soil compaction and whole-tree harvesting plus forest floor removal (referred to as FFR below, compared with stem-only harvesting) on available N, microbial biomass C (MBC), microbial biomass N (MBN), and microbial biomass P (MBP), and dehydrogenase, protease, and phosphatase activities in the forest floor and 0–10 cm mineral soil in a boreal aspen (Populus tremuloides Michx.) forest soil near Dawson Creek, British Columbia, Canada. In the forest floor, no soil compaction effects were observed for any of the soil microbial or enzyme activity parameters measured. In the mineral soil, compaction reduced available N, MBP, and acid phosphatase by 53, 47, and 48%, respectively, when forest floor was intact, and protease and alkaline phosphatase activities by 28 and 27%, respectively, regardless of FFR. Forest floor removal reduced available P, MBC, MBN, and protease and alkaline phosphatase activities by 38, 46, 49, 25, and 45%, respectively, regardless of soil compaction, and available N, MBP, and acid phosphatase activity by 52, 50, and 39%, respectively, in the noncompacted soil. Neither soil compaction nor FFR affected dehydrogenase activities. Reductions in microbial biomass and protease and phosphatase activities after compaction and FFR likely led to the reduced N and P availabilities in the soil. Our results indicate that microbial biomass and enzyme activities were sensitive to soil compaction and FFR and that such disturbances had negative consequences for forest soil N and P cycling and fertility.     

Influence of non-cellulose structural carbohydrate composition on plant material decomposition in soil

The C mineralisation pattern during the early stage of decomposition of plant materials is largely determined by their content of different carbohydrates. This study investigated whether detailed plant analysis could provide a better prediction of C mineralisation during decomposition than proximate analysis [neutral detergent solution (NDF)/acid detergent solution (ADF)]. The detailed analysis included sugars, fructans, starch, pectin, cellulose, lignin and organic N. To determine whether differences in decomposition rate were related to differences in hemicellulose composition, the analysis particularly emphasised the concentrations of arabinose and xylose in hemicelluloses. Carbon dioxide evolution was monitored hourly in soil amended with ten different plant materials. Principal component and regression analysis showed that C mineralisation during day 1 was closely related to free sugars, fructans and soluble organic N components (R ² = 0.83). The sum of non-cellulose structural carbohydrates (intermediate NDF/ADF fraction) was not related to C mineralisation between days 1 and 9. In contrast, a model including starch and protein in addition to the non-cellulose structural carbohydrates, with the hemicelluloses replaced by arabinose and xylose, showed a strong relationship with evolved CO₂ (R ² = 0.87). Carbon mineralisation between days 9 and 34 was better explained by xylan, cellulose and lignin (R ² = 0.72) than by lignocellulose in the ADF fraction. Our results indicated that proximate analyses were not sufficient to explain differences in decomposition. To predict C mineralisation from the range of plant materials studied, we propose a minimum set of analyses comprising total N, free sugars, starch, arabinose, xylan, cellulose and lignin.     

Characterization of the morphological,physical, and mechanical properties of seven nonwood plant fiber bundles

The morphological, physical, and mechanical properties of the nonwood plant fiber bundles of ramie, pineapple, sansevieria, kenaf, abaca, sisal, and coconut fiber bundles were investigated. All fibers except those of coconut fiber had noncircular cross-sectional shapes. The crosssectional area of the fiber bundles was evaluated by an improved method using scanning electron microscope images. The coefficient factor defined as the ratio of the cross-sectional area determined by diameter measurement, to the cross-sectional area determined by image analysis was between 0.92 and 0.96 for all fibers. This indicated that the area determined by diameter measurement was available. The densities of the fiber bundles decreased with increasing diameters. The diameters of each fiber species had small variation of around 3.4%-9.8% within a specimen. The tensile strength and Young’s modulus of ramie, pineapple, and sansevieria fiber bundles showed excellent values in comparison with the other fibers. The tensile strength and Young’s modulus showed a decreasing trend with increasing diameter of fiber bundles.