Characterization of Subtilosin Gene In Wild Type Bacillus Spp. And Possible Physiological Role

In a designed study to screen for antimicrobial exhibiting bacteria using molecular aspects, Bacillus species were considered to investigate antibiotic biosynthesis genes. 28 bacterial strains and 3 induced mutants were screened for the presence of subtilosin gene (sbo) and subtilosin through PCR and Mass spectrometry respectively. Sbo gene was detected in 16 out of 28 Bacillus strains. The results from gene sequences deliberated by multiple sequence alignments revealed high-level homology to the sequences of the sbo-alb gene locus of B. subtilis 168 and the other limited reported strains. Hence, this report provided additional strains to support the idea of subtilosin gene predominance amongst Bacillus strains isolated from environment and to find different species containing homologous genes, furthermore the utilization of its conserved region as a means of identifying Bacillus spp. that produce subtilosin. This is the first report to confirm the detection of subtilosin production from B. amyloliquefaciens.

1H NMR-Based Chemometrics to Gain Insights Into the Bran of Radiation-Induced Colored Wheat Mutant

Recently, wheat has attracted attention as a functional food, rather than a simple dietary energy source. Accordingly, whole-grain intake increases with an understanding of bioactive phytochemicals in bran. The development of colored wheat has drawn more attention to the value of bran owing to its nutritional quality, as well as the antioxidant properties of the colorant. The present 1H NMR-based chemometric study evaluated the compositional improvement of radiation-induced mutants in purple wheat by focusing on the predominant metabolites with high polarity. A total of 33 metabolites, including three choline derivatives, three sugar alcohols, four sugars, 13 amino acids, eight organic acids, and two nucleosides, were identified throughout the 1H NMR spectra, and quantification data were obtained for the identified metabolites via peak shape-based quantification. Principal component and hierarchical cluster analyses were conducted for performing multivariate analyses. The colored original wheat was found to exhibit improvements compared to yellow wheat in terms of the contents of primary metabolites, thus highlighting the importance of conducting investigations of polar metabolites. The chemometrics studies further revealed mutant lines with a compositional enhancement for metabolites, including lysine, proline, acetate, and glycerol.

The repair of topoisomerase 2 cleavage complexes in Arabidopsis

DNA–protein crosslinks (DPCs) and DNA double-stranded breaks (DSBs), including those produced by stalled topoisomerase 2 cleavage complexes (TOP2ccs), must be repaired to ensure genome stability. The basic mechanisms of TOP2cc repair have been characterized in other eukaryotes, but we lack information for plants. Using CRISPR/Cas-induced mutants, we show that Arabidopsis thaliana has two main TOP2cc repair pathways: one is defined by TYROSYL-DNA-PHOSPHODIESTERASE 2 (TDP2), which hydrolyzes TOP2–DNA linkages, the other by the DNA-dependent protease WSS1A (a homolog of human SPARTAN/yeast weak suppressor of smt3 [Wss1]), which also functions in DPC repair. TDP1 and TDP2 function nonredundantly in TOP1cc repair, indicating that they act specifically on their respective stalled cleavage complexes. The nuclease METHYL METHANESULFONATE AND UV-SENSITIVE PROTEIN 81 (MUS81) plays a major role in global DPC repair and a minor role in TOP2cc repair. DSBs arise as intermediates of TOP2cc repair and are repaired by classical and alternative nonhomologous end joining (NHEJ) pathways. Double-mutant analysis indicates that “clean” DNA ends caused by TDP2 hydrolysis are mainly religated by classical NHEJ, which helps avoid mutation. In contrast, the mutagenic alternative NHEJ pathway mainly processes nonligateable DNA ends. Thus, TDP2 promotes maintenance of plant genome integrity by error-free repair of TOP2cc.

Comparative transcriptome analyses shed light on carotenoid production and plastid development in melon fruit

Carotenoids, such as β-carotene, accumulate in chromoplasts of various fleshy fruits, awarding them with colors, aromas, and nutrients. The Orange (CmOr) gene controls β-carotene accumulation in melon fruit by posttranslationally enhancing carotenogenesis and repressing β-carotene turnover in chromoplasts. Carotenoid isomerase (CRTISO) isomerizes yellow prolycopene into red lycopene, a prerequisite for further metabolism into β-carotene. We comparatively analyzed the developing fruit transcriptomes of orange-colored melon and its two isogenic EMS-induced mutants, low-β (Cmor) and yofi (Cmcrtiso). The Cmor mutation in low-β caused a major transcriptomic change in the mature fruit. In contrast, the Cmcrtiso mutation in yofi significantly changed the transcriptome only in early fruit developmental stages. These findings indicate that melon fruit transcriptome is primarily altered by changes in carotenoid metabolic flux and plastid conversion, but minimally by carotenoid composition in the ripe fruit. Clustering of the differentially expressed genes into functional groups revealed an association between fruit carotenoid metabolic flux with the maintenance of the photosynthetic apparatus in fruit chloroplasts. Moreover, large numbers of thylakoid localized photosynthetic genes were differentially expressed in low-β. CmOR family proteins were found to physically interact with light-harvesting chlorophyll a–b binding proteins, suggesting a new role of CmOR for chloroplast maintenance in melon fruit. This study brings more insights into the cellular and metabolic processes associated with fruit carotenoid accumulation in melon fruit and reveals a new maintenance mechanism of the photosynthetic apparatus for plastid development.

Mutation breeding in six rowed barley: Screening for stem rust resistance in certain induced mutants

During the induced mutagenesis in six rowed barley, 18 mutant lines were isolated from the M2 and M3 generations. Thirteen lines were induced by gamma rays, four by EMS and one by HA. These lines were screened for resistance to stem rust and loss assessment. Mutants were grown in field as well as in earthen pots. Artificial inoculums were provided in condition suitable for rust occurrence. Rust reaction was recorded according to modified Cabb’s scale and the rust intensity was classified as 0,5, 10, 25, 40 and 65 percent. Lines K50-38, K50-97, K50-102 and K40-73 were completely free from infection (0 %) and thought to be immune lines(I). K50-47, KE-4-47 and G50-47 exhibited traces of symptoms and placed under 5% intensity and were very resistant(R). Line KE2-63 exhibited 10% intensity and classified as moderately resistant(MR). Line K50-44; K40-11, K40-67, K30-91, KE3-75, G50-35 and GH3-32 showed 25% intensity of pustules and were put into moderately susceptible(MS) class. One line K30-82 showed 40% intensity and classified as susceptible(S), while line KE4-87 exihibited 65% intensity and was classified as heavily susceptible. Class 100 percent was not observed. Immune, very resistant and moderately resistant were identified as resistant lines and selected for further study, while moderately susceptible and heavily susceptible lines were identified as susceptible lines and discarded. Reduction in yield contributing traits i.e. number of grains/spike, test weight, seed yield and biological yield was also observed which was positively correlated with disease intensity.

COMMUNICATION SYSTEMS OF TOPOLS IN COMMUNICATION WITH SYSTEMS OF THEIR REPRODUCTION

We consider poplar selection systems in connection with their reproduction systems. Poplar is a model object on which it is possible to test all known systems for the selection of woody plants. Positive selection and synthetic selection methods (hybridization, mutagenesis, polyploidy, genetic engineering, biotechnology) effectively “work” on poplar. Poplar is propagated by seed, vegetative and in vitro biotechnology. Amphimixis or seed propagation of poplar is used to create hybrid seed plantations, for example white poplar × aspen. In natural poplar populations, spontaneous hybridization occurs, leading to the emergence of new species. The ability to vegetatively propagate poplar successfully solves the problem of plus breeding: selected best biotypes are propagated by in vitro grafting, cuttings or regenerants, and plantation cultures are created. Synthetic breeding methods: hybridization is widely used on poplar; dozens of poplar hybrids for forest crops and landscaping have been obtained; Induced mutants and polyploids are obtained. Breeders get their first genetically modified forms using genetic engineering. By microcloning, economically valuable individuals are propagated and regenerants are used to create forest crops and landscaping.