Primerjalna študija genetske raznovrstnosti talnih bakterij in gliv v različnih sukcesijah vegetacije na krasu v provinci Guizhou, Kitajska

To study the soil genetic diversity of bacteria and fungi in different vegetation successions (grassland, shrubbery, primary forest and secondary forest) from the karst area, the Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) technology was applied. The results showed that: (1) the diversity of bacterial communities and the fungal communities in karst area were higher than non karst area in each vegetation succession. Compared with the survey from bacterial (the Shannon index was 2.97 in primary forest, 2.91 in secondary forest, 3.18 in shrubbery, 3.14 in grassland and 2.68 in non karst), fungal diversity between karst areas (the Shannon index was 3.56 in primary forest, 3.78 in secondary forest, 3.73 in shrubbery and 3.70 in grassland) and non karst areas (the Shannon index was 3.08) was more evident, which may be related to the alterations of the composition of plant community and the source of carbon in soil with the vegetation succession of karst ecosystem; (2) The comparation of bacterial diversity index and the richness comprehensively evaluated as follows: shrubbery > grassland > primary forest > nsecondary forest. The diversity index and the richness of fungal communities was as follows: secondary forest > shrubbery > grassland > primary forest. The results suggest that the fungal communities have been greatly changed via vegetation successions, but the diversity index and the richness of the bacterial communities have not been seriously affected. The results provide scientific basis for understanding karst surface ecosystem, which contributes to the future aim of protecting the karst from desertification.Za proučevanje genetske pestrosti talnih bakterij in gliv v različnih sukcesijah vegetacije (travišče, grmičevje, primarni gozd in sekundarni gozd) na krasu je bila uporabljena tehnologija verižne reakcije s polimerazo-denaturirajoča gradientna gelska elektroforeza (PCR-DGGE). Rezultati raziskave so pokazali, da: (1) je bila v vsaki sukcesiji vegetacije pestrost bakterijskih in glivnih združb na kraškem območju višja kot na nekraškem. V primerjavi z bakterijsko raznovrstnostjo (Shannonov indeks je bil 2,97 v primarnem gozdu, 2,91 v sekundarnem gozdu, 3,18 v tleh grmičevja, 3,14 v tleh travišč in 2,68 v nekraškem območju) je bila raznovrstnost gliv med kraškimi območji (Shannonov indeks je bil v primarnem gozdu 3,56, 3,78 v sekundarnem gozdu, 3,73 v tleh grmičevja in 3,70 v tleh travišč) in nekraškimi (Shannonov indeks je bil 3,08) jasneje izražena. To je lahko povezano s spremembami v sestavi rastlinske združbe in vira ogljika v tleh glede na stanje sukcesije v vegetaciji kraškega ekosistema. (2) Primerjava kazalnikov bakterijske raznovrstnosti in abundance je bila celostno ovrednotena in sledi takole: grmičevje > travišče > primarni gozd > sekundarni gozd. Kazalnika raznovrstnosti in abundance glivnih združb kažeta sledeči trend: sekundarni gozd > grmičevje > travišče > primarni gozd. Rezultati izkazujejo, da so se glivne združbe precej spremenile zaradi sukcesije v vegetaciji, vendar pa na drugi strani ni bilo bistvenega vpliva na kazalnika bakterijske raznovrstnosti in abundance. Rezultati med drugim dajejo tudi znanstveno podlago za razumevanje delovanja kraškega površinskega ekosistema, kar ključno prispeva k cilju zaščite krasa pred dezertifikacijo (širjenjem puščav)

Preferential Localization of Human Origins of DNA Replication at the 5′-Ends of Expressed Genes and at Evolutionarily Conserved DNA Sequences

Replication of mammalian genomes requires the activation of thousands of origins which are both spatially and temporally regulated by as yet unknown mechanisms. At the most fundamental level, our knowledge about the distribution pattern of origins in each of the chromosomes, among different cell types, and whether the physiological state of the cells alters this distribution is at present very limited.We have used standard λ-exonuclease resistant nascent DNA preparations in the size range of 0.7–1.5 kb obtained from the breast cancer cell line MCF–7 hybridized to a custom tiling array containing 50–60 nt probes evenly distributed among genic and non-genic regions covering about 1% of the human genome. A similar DNA preparation was used for high-throughput DNA sequencing. Array experiments were also performed with DNA obtained from BT-474 and H520 cell lines. By determining the sites showing nascent DNA enrichment, we have localized several thousand origins of DNA replication. Our major findings are: (a) both array and DNA sequencing assay methods produced essentially the same origin distribution profile; (b) origin distribution is largely conserved (>70%) in all cell lines tested; (c) origins are enriched at the 5′ends of expressed genes and at evolutionarily conserved intergenic sequences; and (d) ChIP on chip experiments in MCF-7 showed an enrichment of H3K4Me3 and RNA Polymerase II chromatin binding sites at origins of DNA replication.Our results suggest that the program for origin activation is largely conserved among different cell types. Also, our work supports recent studies connecting transcription initiation with replication, and in addition suggests that evolutionarily conserved intergenic sequences have the potential to participate in origin selection. Overall, our observations suggest that replication origin selection is a stochastic process significantly dependent upon local accessibility to replication factors

Influence of arbuscular mycorrhizal fungi on mercury accumulation in rice (Oryza sativa L.): From enriched isotope tracing perspective

The microorganisms that co-exist between soil and rice systems in heavy metal-contaminated soil environments play important roles in the heavy metal pollution states of rice, as well as in the growth of the rice itself. In this study, in order to further examine the effects of soil microorganisms on the mercury (Hg) uptake of rice plants and determine potential soil phytoremediation agents, an enriched 199Hg isotope was spiked in a series of pot experiments to trace the absorption and migration of Hg and rice growth in the presence of arbuscular mycorrhizal fungi (AMF). It was observed that the AMF inoculations significantly reduced the Hg concentration in the rice. The Hg concentration in rice in the AMF inoculation group was between 52.82% and 96.42% lower than that in the AMF non-inoculation group. It was also interesting to note that the presence of AMF tended to cause Hg (especially methyl-Hg (Me199Hg)) to migrate and accumulate in the non-edible parts of the rice, such as the stems and leaves. Under the experimental conditions selected in this study, the proportion of Me199Hg in rice grains decreased from 9.91% to 27.88%. For example, when the exogenous Hg concentration was 0.1 mg/kg, the accumulated methyl-Hg content in the grains of the rice in the AMF inoculation group accounted for only 20.19% of the Me199Hg content in the rice plants, which was significantly lower than that observed in the AMF non-inoculated group (48.07%). AMF also inhibited the absorption of Hg by rice plants, and the decrease in the Hg concentration levels in rice resulted in significant improvements in growth indices, including biomass and micro-indexes, such as antioxidant enzyme activities. The improvements occurred mainly because the AMF formed symbiotic structures with the roots of rice plants, which fixed Hg in the soil. AMF also reduce the bioavailability of Hg by secreting a series of substances and changing the physicochemical properties of the rhizosphere soil. These findings suggest the possibility of using typical co-existing microorganisms for the remediation of soil heavy metal contamination and provide valuable insights into reducing human Hg exposure through rice consumption

Next-generation transgenic cotton: pyramiding RNAi and Bt counters insect resistance

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are extensively cultivated worldwide. To counter rapidly increasing pest resistance to crops that produce single Bt toxins, transgenic plant 'pyramids' producing two or more Bt toxins that kill the same pest have been widely adopted. However, cross-resistance and antagonism between Bt toxins limit the sustainability of this approach. Here we describe development and testing of the first pyramids of cotton combining protection from a Bt toxin and RNA interference (RNAi). We developed two types of transgenic cotton plants producing double-stranded RNA (dsRNA) from the global lepidopteran pest Helicoverpa armigera designed to interfere with its metabolism of juvenile hormone (JH). We focused on suppression of JH acid methyltransferase (JHAMT), which is crucial for JH synthesis, and JH-binding protein (JHBP), which transports JH to organs. In 2015 and 2016, we tested larvae from a Bt-resistant strain and a related susceptible strain of H. armigera on seven types of cotton: two controls, Bt cotton, two types of RNAi cotton (targeting JHAMT or JHBP) and two pyramids (Bt cotton plus each type of RNAi). Both types of RNAi cotton were effective against Bt-resistant insects. Bt cotton and RNAi acted independently against the susceptible strain. In computer simulations of conditions in northern China, where millions of farmers grow Bt cotton as well as abundant non-transgenic host plants of H. armigera, pyramided cotton combining a Bt toxin and RNAi substantially delayed resistance relative to using Bt cotton alone

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field