Over half of all organisms living on Earth today are parasites, and there are hardly any species which are free from them. How can the “wild immunology” approach help us understand them better?
The whitefly, Bemisia tabaci, an insect of the order Hemiptera which attacks more than 600 species of plants, is one of the most important agricultural pests around the world. The insecticidal Cry proteins from Bacillus thuringiensis (Bt) are useful biological pesticides, and some are toxic to Hemipteran insects. In this study, Colombian native isolates of Bt were functionally characterized at molecular and biological levels. The strains contained between one and five different crystal shapes: round, triangular, amorphous, bipyramidal and squared. The strains presented between three to seven bands of proteins in their electrophoretic pattern that were organized into six groups according to their possible biological activity on insect pests. Cry1Aa, cry1Ab, cry1Ac, cry1B and cry1C genes were identified for PCR in the different Bt isolates. Bioassays were performed on tomato leaves whose surface was spread with 3 μg · ml−1 crude extract of Bt toxins. Second instar larvae of whitefly, which were placed on top of leaves and exposed to the toxins for 7 days, exhibited mortalities from 18 to 69%. The lethal concentration 50 (LC50) of ZBUJTL39, Bt kurstaki HD1 and ZCUJTL9 strains were 1.83, 1.85 and 2.16 μg · ml−1, respectively (p < 0.05). These results show that the native Bt strain ZBUJTL39, which contained the genes cry1Aa, cry1Ab, cryCa and cryBa could eventually be used for the development of an integrated management program together with other tools for the control of B. tabaci.
Gene postulation is one of the fastest and most cost-effective methods for identifying seedling leaf rust resistance genes in wheat cultivars. Many researchers use this approach to identify Lr genes in wheat cultivars. The purpose of our research was to identify seedling leaf rust resistance genes in 20 wheat cultivars from different breeding centers of Russia, Ukraine and Germany. Forty-two near isogenic Thatcher lines and 10 Puccinia triticina isolates were used for gene postulation. When assessing the infection types to cultivars and lines, a scale was used, according to Oelke and Kolmer. In 20 wheat cultivars 19 Lr genes were postulated: 2c, 3, 10, 3bg, 3ka, 14a, 17, 18, 23, 25, 26, 30, 33, 40, 44, 50, B, Exch, Kanred. The most common for cultivars was the Lr10 gene. In five cultivars, showing high field resistance, most postulated seedling genes (Lr2c, Lr3, Lr10, Lr14а, Lr26, Lr33) were not effective in the adult stage. It is possible that resistance of such cultivars is associated with APR genes, the postulation of which requires an expansion in the number and spectrum of P. triticina isolate virulence. Most of the studied cultivars (60%) have recently been entered into the register (2015–2019) and in the field show a stable or moderately susceptible response to P. triticina infection, despite the fact that the Lr genes postulated in them were not effective in the adult stage. The data obtained indicated a variety of genotypes of the studied cultivars, as well as the tendency of breeders to use the effect of pyramiding ineffective genes, which can prolong the resistance of the cultivar. Annual monitoring of varieties is necessary in each region, especially when reacting with a medium susceptible type (MS), which may indicate the initial stage of resistance loss.
The aim of the study was to develop a reliable and cost-effective method for detection of nonsense mutation in APAF1 gene causing lethal effect called HH1 (Holstein Haplotype1) and to evaluate its prevalence in a sample of Polish Holstein-Friesian bulls. One hundred seventy eight bulls born between 1996 and 2017 were included in the analysis. They were kept in four artificial insemination centers and have in the pedigree the known carrier of HH1. All bulls were diagnosed by novel PCR-SSCP technique. Specific amplicons of 261 bp APAF1 gene fragment were used to detect changes in single stranded conformation (SSCP) caused by nonsense mutation C/T responsible for HH1. Each new carrier was used to trace another potential carriers among their offspring available in Polish Holstein Bull Repository Database. Among 178 bulls, 85 HH1 carriers were found. Our results show that nonsense mutation in APAF1 gene is already transmitted and segregating in Polish Holstein-Friesian cattle and its frequency may increase if no action will be undertaken against actual carriers.
In our previous Genome-wise Association Study we found that Cystic Fibrosis Transmem- brane Conductance Regulator gene (CFTR) is a candidate gene for sperm motility in fresh semen of Holstein-Friesian bulls. Since in cows thawed semen is commonly used for the artificial insem- ination (AI) we have decided to find out whether functional polymorphism within CFTR gene coding sequence is associated with selected parameters of thawed sperm, including their motility evaluated by computer-assisted sperm analysis (CASA), the activity of three antioxidant enzymes: glutathione peroxidase (GPx) catalase (CAT), superoxide dismutase (SOD), ATP con- tent and integrity of sperm membranes. One hundred twenty Holstein Friesian bulls kept in uni- form environmental conditions (one AI company) were included in the study. Significant associ- ations between genotypes of missense mutation within exon 11 of the CFTR gene (Met468Leu) and the activity of antioxidant enzymes and sperm mitochondrial function were revealed. No effect of CFTR genotypes on sperm motility was observed. Significant differences in CAT and SOD activity were found between AA and TT homozygous individuals. Bulls with TT genotype had the lowest activity of both antioxidant enzymes. The same bulls also showed the lowest num- ber of sperm with active mitochondria. Our results demonstrate that missense mutation Met468Leu within CFTR gene is associated with antioxidant enzyme activity and mitochondrial function of bovine thawed sperm without affecting their motility.
The aim of the study was to find out whether carriers of new lethal mutation in SDE2 gene occur in the population of Polish Holstein-Friesian bulls. Eighty seven bulls were included in the analysis. Bulls were selected as having in the pedigree known carrier of SDE2 mutation (bull Mountain USAM000002070579). All bulls were diagnosed by PCR amplification of 524 bp fragment of SDE2 gene followed by digestion of Bcc I restriction enzyme. Heterozygotes (carriers) were confirmed by sequencing. Each new carrier was used to trace another potential carriers among its offspring available in Polish Holstein Bull Repository Database. Among 87 bulls, 50 new SDE2 carriers were found. The study has shown that mutation in SDE2 gene causing early embryo mortality is already transmitted to Polish Holstein-Friesian cattle. The results are sufficient to initiate the screening program to reveal new carriers and to avoid further spreading of SDE2 lethal mutation.
Clarifying the genetic background of the drought-tolerance trait is a crucial task that may help to improve plant performance under stress by a genetic engineering approach. Dehydration-responsive element-binding protein (DREB) is a transcription factor family which modulates many stress-responsive genes. In this study, we isolated a DREB homolog gene named ZmDREBtv from Zea mays var. Tevang-1. Using bioinformatic tools, a number of InDels and SNPs in ZmDREBtv sequence different from the reference accession were identified. In addition, based on deduced protein sequence similarity, ZmDREBtv was assigned to transcription factor DREB2 class as featured by a conserved DNA binding domain - AP2. The ZmDREBtv construct under thecontrol of the rd29A promoter was transformed into a drought-sensitive maize plant, K7 line. The transgenic plants were assessed with reference to molecular and phenotypic characteristics related to the drought-tolenrance trait. The results proved that the maize plants carrying ZmDREBtv gene showed enhanced tolerance and better performance to the water-deficit environment at different stages, compared to the wild-type plants.
Cells of a multicellular organism are genetically identical but differ in structure and function. This heterogeneity is created by several epigenetic mechanisms during the development of the organism. The epigenetic changes- including DNA methylation, histone post-translational modifications, chromatin remodeling and RNA interference have all been shown to control chromatin structure and regulate a plethora of cellular and organismal processes. There is a strong evidence that epigenetics play a crucial role in the development of diseases such as cancer, schizophrenia or metabolic disorders. The epigenetic regulation underlie memory formation or adaptation to external stimuli. The extent to which environmental effects can provoke epigenetic responses represents an exciting area of future research. Here we review the current knowledge about the epigenetic mechanisms and their relation to the human health and disease.