Functional analysis of the sortase YhcS in Bacillus subtilis

Sortases of Gram-positive bacteria catalyze the covalent C-terminal anchoring of proteins to the cell wall. Bacillus subtilis, a well-known host organism for protein production, contains two putative sortases named YhcS and YwpE. The present studies were aimed at investigating the possible sortase function of these proteins in B. subtilis. Proteomics analyses revealed that sortase-mutant cells released elevated levels of the putative sortase substrate YfkN into the culture medium upon phosphate starvation. The results indicate that YfkN required sortase activity of YhcS for retention in the cell wall. To analyze sortase function in more detail, we focused attention on the potential sortase substrate YhcR, which is co-expressed with the sortase YhcS. Our results showed that the sortase recognition and cell-wall-anchoring motif of YhcR is functional when fused to the Bacillus pumilus chitinase ChiS, a readily detectable reporter protein that is normally secreted. The ChiS fusion protein is displayed at the cell wall surface when YhcS is co-expressed. In the absence of YhcS, or when no cell-wall-anchoring motif is fused to ChiS, the ChiS accumulates predominately in the culture medium. Taken together, these novel findings show that B. subtilis has a functional sortase for anchoring proteins to the cell wall.     

Effect of endophytic fungi on cadmium tolerance and bioaccumulation by Festuca arundinacea and Festuca pratensis

Endophytic fungi are a group of fungi that live asymptomatically inside plant tissue. These fungi may increase host plant tolerance to biotic and abiotic stresses. The effect of Neotyphodium endophytes in two grass species (Festuca arundinacea and Festuca pratensis) on cadmium (Cd) tolerance, accumulation and translocation has been our main objective. The plants were grown in a hydroponic system under different Cd concentrations (0, 5, 10, and 20 mg L(-1)) for 6 weeks. They were also grown in soil spiked with different concentrations of Cd (0, 10, 20, and 40 mg kg(-1)) for 2 months. The results from all Cd treatments showed higher biomass production (12-24%) and higher potential to accumulate Cd in roots (6-16%) and shoots (6-20%) of endophyte-infected plants than endophyte-free plants. Cadmium accumulation by plants indicated that the grasses were capable of Cd hyperaccumulation, a property that was augmented after endophyte infection. Maximum photochemical efficiency of photosystem II (Fv/Fm) revealed that Cd stress was significantly reduced in endophyte-infected plants compared to non-infected ones.     

Study of genetic variation in sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) using agro-morphological traits and <Emphasis Type="Italic">ISSR</Emphasis> markers

This research was conducted to study the genetic variation among eighteen genotypes of sesame (Sesamum indicum L.) collected from various agro-climatic regions of Iran along with six exotic genotypes from the Asian countries using both agro-morphological and ISSR marker traits. The results showed significant differences among genotypes for all agro-morphological traits and a relatively high genetic coefficient of variation observed for number of fruiting branches per plant, capsules per plant, plant height and seed yield per plant. Cluster analysis based on these traits grouped the genotypes into five separate clusters. Larger interthan intra cluster distances implies the presence of higher genetic variability between the genotypes of different groups. Genotypes of two clusters with a good amount of genetic divergence and desirable agronomic traits were detected as promising genotypes for hybridization programs. The 13 ISSR primers chosen for molecular analysis revealed 170 bands, of which 130 (76.47%) were polymorphic. The generated dendrogram based on ISSR profiles divided the genotypes into seven groups. A principal coordinate analysis confirmed the results of clustering. The agro-morphological traits and ISSR markers reflected different aspects of genetic variation among the genotypes as revealed by a non significant cophenetic correlation in the Mantel test. Therefore the complementary application of both types of information is recommended to maximize the efficiency of sesame breeding programs. The discordance among diversity patterns and geographical distribution of genotypes found in this investigation implies that the parental lines for hybridization should be selected based on genetic diversity rather than the geographical distribution.     

Evolution of Carthamus species revealed through sequence analyses of the fad2 gene family

Physiology and Molecular Biology of Plants - The diversity of 11 fatty acid desaturase (fad2) genes has not been investigated between cultivated and wild species in&nbsp;the Carthamus genus. In...     

Mealybug, Phenococcus solani, and barley aphid, Sipha maydis, response to endophyte-infected tall and meadow fescues

Mealybugs and aphids are insects which damage grass species. The effects of fungal endophytes on the feeding of the mealybug, Phenococcus solani Ferris (Homoptera: Pseudococcidae), and barley aphid, Sipha maydis Passerini (Homoptera: Aphididae), on tall fescue, Festuca arundinacea Schreb. and meadow fescue, Festuca pratensis Huds., were studied under greenhouse conditions. Mealybugs preferred endophyte‐free (E–) clones over their endophyte‐infected (E+) counterparts. E+ plants had a significantly lower number of mealybugs than E– plants. A mixture of E+ and E– plants supported intermediate mealybug numbers, between pure plantings of E+ and E– grasses. Barley aphids released on to plant materials were deterred from feeding and could not persist on E+ plants. E– plants did not survive because of aphid damage, while E+ plants generally re‐grew, but were damaged to some degree. The results showed that the use of pure stands of endophyte‐infected grasses or a mixed stand of infected and non‐infected plants may increase the persistence and durability of turf and forage grass species in the presence of foliar damaging insects.     

Inheritance of flower color and spininess in safflower (Carthamus tinctorius L.)

Safflower (Carthamus tinctorius L.) flowers are used for coloring and flavoring food and also as fresh-cut and dried flowers. The most important characteristics which contribute to the ornamental value of safflower are flower color and spinelessness. The objective of this study was to determine the inheritance mode and the number of genes controlling spininess and flower color in some Iranian genotypes of safflower. The results indicated that the existence of spines on the leaves and bracts of safflower is controlled by a single dominant gene in which the spiny phenotype was completely dominant to spineless. In some crosses, flower color was controlled by two epistatic loci each with two alleles, resulting in a ratio of 13:3 in the segregating F2 population for plants with orange and yellow flowers. Also, other mechanisms of genetic control, such as duplicate dominance and duplicate recessive types of epistasis, were observed for flower color in other crosses that led to ratios of 7:9 and 15:1 for plants with orange and yellow flowers, respectively. The results suggest that for ornamental use or in the food dying industry, genotypes with orange or yellow flowers and without spines on the leaves and bracts can be produced.     

Cytogenetic and crossability studies in hulled wheat collected from Central Zagros in Iran

Underutilized hulled wheat species may be valuable sources of genes for wheat improvement. Hulled wheats of mountainous Central Zagros area of Iran are poorly studied with unclear classification status. In this study, the region was extensively searched and hulled wheats were sampled from local farmers’ fields. Cytogenetic studies of several samples revealed that these wheats are tetraploid with a chromosome number of 2n = 28. Considerable level of karyotypic resemblance was detected between these hulled wheats and durum wheat genotypes, suggesting their genomic similarity. To compare the degree of cross compatibility, F1 progenies were produced using three durum accessions and a bread wheat cultivar as female parent in crosses with two of the collected hulled wheat samples, namely Zarneh and Jonghan. The highest crossability rate was observed when Aria, a durum wheat cultivar, was used as female parent. In general, Jonghan showed higher crossability rate with both bread and durum cultivars as compared to Zarneh. The meiotic behavior of F1 genotypes was analyzed to determine genomic characteristics of hulled wheat samples. Microscopic examination of pollen mother cells at meiotic metaphase strongly suggested the AABB type genome for the hulled wheat samples. Meiotic behaviors of hybrids derived from cross with Zarneh as male parent showed more abnormalities than the other male parent. Meiotic restitution, chromosomes laggards and chromosomal bridges were noticed and the extent varied for each cross. It may be concluded that the hulled wheats of Central Zagros that are currently cultivated are mainly tetraploids emmer containing AABB genome and are crossable with durum wheat, producing hybrids mostly with normal meiotic behaviors.     

Widespread occurrence of Neotyphodium-like endophyte in populations of Bromus tomentellus Boiss. in Iran

Neotyphodium species occur as endophytic fungi in cool-season grasses around the world. The beneficial aspects of grass-Neotyphodium associations have provoked researchers to look for a novel association in plant species where this symbiotum has not been reported. We surveyed Russian bromegrass (Bromus tomentellus Boiss.) accessions from a germplasm collection for the presence of Neotyphodium spp. fungi and determined levels of endophyte infection in B. tomentellus populations in native rangelands of Iran. Among 50 collected accessions, symbiotic fungi were detected in 45 accessions without any symptoms of choke disease on host plants. In culture medium, fast-growing endophytes appeared from seeds after 7-14 days. Plants grown from seed collections were 80-100% infected. Based on morphological characteristics and PCR analysis, we concluded that this fungus is a member of the Neotyphodium group of endophytic fungi. Lack of apparent toxicity to grazing animals suggests a place for endophyte-infected B. tomentellus in rangeland renovation, providing this infected grass exhibits increased tolerance to abiotic stresses.     

Study of genetic variation in sesame (Sesamum indicum L.) using agro-morphological traits and ISSR markers

This research was conducted to study the genetic variation among eighteen genotypes of sesame (Sesamum indicum L.) collected from various agro-climatic regions of Iran along with six exotic genotypes from the Asian countries using both agro-morphological and ISSR marker traits. The results showed significant differences among genotypes for all agro-morphological traits and a relatively high genetic coefficient of variation observed for number of fruiting branches per plant, capsules per plant, plant height and seed yield per plant. Cluster analysis based on these traits grouped the genotypes into five separate clusters. Larger inter- than intra cluster distances implies the presence of higher genetic variability between the genotypes of different groups. Genotypes of two clusters with a good amount of genetic divergence and desirable agronomic traits were detected as promising genotypes for hybridization programs. The 13 ISSR primers chosen for molecular analysis revealed 170 bands, of which 130 (76.47%) were polymorphic. The generated dendrogram based on ISSR profiles divided the genotypes into seven groups. A principal coordinate analysis confirmed the results of clustering. The agro-morphological traits and ISSR markers reflected different aspects of genetic variation among the genotypes as revealed by a non significant cophenetic correlation in the Mantel test. Therefore the complementary application of both types of information is recommended to maximize the efficiency of sesame breeding programs. The discordance among diversity patterns and geographical distribution of genotypes found in this investigation implies that the parental lines for hybridization should be selected based on genetic diversity rather than the geographical distribution.