Phosphorylation and ubiquitination of dynamin-related proteins (AtDRP3A/3B) synergically regulate mitochondrial proliferation during mitosis

The balance between mitochondrial fission and fusion is disrupted during mitosis, but the mechanism governing this phenomenon in plant cells remains enigmatic. Here, we used mitochondrial matrix‐localized Kaede protein (mt‐Kaede) to analyze the dynamics of mitochondrial fission in BY‐2 suspension cells. Analysis of the photoactivatable fluorescence of mt‐Kaede suggested that the fission process is dominant during mitosis. This finding was confirmed by an electron microscopic analysis of the size distribution of mitochondria in BY‐2 suspension cells at various stages. Cellular proteins interacting with Myc‐tagged dynamin‐related protein 3A/3B (AtDRP3A and AtDRP3B) were immunoprecipitated with anti‐Myc antibody‐conjugated beads and subsequently identified by microcapillary liquid chromatography–quadrupole time‐of‐flight mass spectrometry (CapLC Q‐TOF) MS/MS. The identified proteins were broadly associated with cytoskeletal (microtubular), phosphorylation, or ubiquitination functions. Mitotic phosphorylation of AtDRP3A/AtDRP3B and mitochondrial fission at metaphase were inhibited by treatment of the cells with a CdkB/cyclin B inhibitor or a serine/threonine protein kinase inhibitor. The fate of AtDRP3A/3B during the cell cycle was followed by time‐lapse imaging of the fluorescence of Dendra2‐tagged AtDRP3A/3B after green‐to‐red photoconversion; this experiment showed that AtDRP3A/3B is partially degraded during interphase. Additionally, we found that microtubules are involved in mitochondrial fission during mitosis, and that mitochondria movement to daughter cell was limited as early as metaphase. Taken together, these findings suggest that mitotic phosphorylation of AtDRP3A/3B promotes mitochondrial fission during plant cell mitosis, and that AtDRP3A/3B is partially degraded at interphase, providing mechanistic insight into the mitochondrial morphological changes associated with cell‐cycle transitions in BY‐2 suspension cells.     

渭河流域大型底栖动物群落结构及其与环境因子的关系

于2011年10月对渭河流域45个采样点的大型底栖动物进行调查采样.共采集到大型底栖动物116属(种),其中水生昆虫91(属)种,占78.4％;软体动物12种,占10.3％;环节动物9种,占7.8％;甲壳动物4种,占3.4％.利用大型底栖动物的物种组成以及物种相对丰度数据,应用双向指示种分析和无偏对应分析将45个样点分为3组.第1组样点的指示物种为锯形蜉属1种(Serratella sp.)、纹石蛾属1种(Hydropsyche sp.)和朝大蚊属1种(Antochasp.);第2组样点指示物种为虻属1种(Tabanus sp.)、Alotanypus venustus、Pelecorhynchidae 1 种、Liodessus sp.和霍甫水丝蚓(Limnodrilus hoffineistteri);第3组指示物种为黑翅蜉(Ephemera nigroptera)和半球多脉扁螺(Polypylis hemisphaerula).典范对应分析表明,卵石+砾石型底质、流速、电导率、水深和总氮显著影响了渭河流域底栖动物群落的空间分布.     

Effects of elevated atmospheric CO2 on soil enzyme activities at different nitrogen application treatments

It has been predicted that elevated atmospheric CO₂ will increase enzyme activity as a result of CO₂-induced carbon entering the soil. The objective of this study was to investigate the effects of elevated atmospheric CO₂ on soil enzyme activities under a rice/wheat rotation. This experiment was conducted in Wuxi, Jiangsu, China as part of the China FACE (Free Air Carbon Dioxide Enrichment) Project. Two atmospheric CO₂ concentrations (580±60) and (380±40) μmol·mol^-1) and three N application treatments (low-150, normal-250 and high-350 kg N·hm^-2) were included. Soil samples (0-10 cm) were collected for analysis of β-glucosidase, invertase, urease, acid phosphates and β-glucosaminidase activities. The results revealed that with elevated atmospheric CO₂ β-glucosidase activity significantly decreased (P < 0.05) at low N application rates; had no significant effect with a normal N application rate; and significantly increased (P < 0.05) with a high N application rate. For urease activity, at low and normal N application rates (but not high N application rate), elevated atmospheric CO₂ significantly increased (P < 0.05) it. With acid phosphatase elevated atmospheric CO₂ only had significant higher effects (P < 0.05) at high N application rates. Under different CO₂ concentration, effects of N fertilization are also different. Soil β-glucosidase activity at ambient CO₂ concentration decreased with N fertilization, while it increased at elevated CO₂ concentration. In addition, invertase and acid phosphatase activities at elevated CO₂ concentration, significantly increased (P < 0.05) with N treatments, but there was no effect with the ambient CO₂ concentration. For urease activity, at ambient CO₂ concentration, N fertilization increased it significantly (P < 0.05), whereas at elevated CO₂ concentration it was not significant. Additionally, with β-glucosaminidase activity, there were no significant effects from N application. In general, then, elevated atmospheric CO₂ increased soil enzyme activity, which may be attributed to the following two factors: (1) elevated atmospheric CO₂ led to more plant biomass in the soil, which in turn stimulated soil microbial biomass and activity; and (2) elevated atmospheric CO₂ increased plant photosynthesis, thereby increasing plant-derived soil enzymes.     

ISSR Analysis of the Genetic Diversity of the Endangered Species Sinopodophyllum hexandrum （Royle） Ying from Western Sichuan Province, China

Slnopodophyllum hexandrum （Royle） Ying Is an Important medicinal and endangered species. Inter-simple sequence repeats （ISSR） analysis was conducted on seven natural populations from western Slchuan Province to Investigate the genetic diversity of S. hexandrum. Leaf samples of 140 Individuals were collected. Of the 139 discernible fragments generated by 12 selected primers （among 100 primers）, 54 appeared to be polymorphlc. The percentage of polymorphlc bands （PPB） was 38.85% at the species level, and PPB within a population ranged from 7.91% to 23.74%. Low levels of genetic variation （He = 0.092, Ho = 0.142） and high levels of genetic differentiation among the populations （Gst= 62.25%） was detected on the basis of results from POPGENE and analysis of molecular variance （AMOVA）, respectively. Furthermore, the limited gene flow （Nm = 0.361） may result from biological characteristics, such as self-pollination and short distance seed dispersal. Based on the genetic and ecological Information available for S. hexandrum, we propose some appropriate strategies for the conservation of the endangered medicinal species in this region, namely rescuing and conserving the core populations for in situ conservation and sampling and preserving more populations with fewer Individuals from each population for ex situ conservation.     

Effects of elevated atmospheric CO2 on soil enzyme activities at different nitrogen application treatments

It has been predicted that elevated atmospheric CO₂ will increase enzyme activity as a result of CO₂-induced carbon entering the soil. The objective of this study was to investigate the effects of elevated atmospheric CO₂ on soil enzyme activities under a rice/wheat rotation. This experiment was conducted in Wuxi, Jiangsu, China as part of the China FACE (Free Air Carbon Dioxide Enrichment) Project. Two atmospheric CO₂ concentrations (580±60) and (380±40) μmol·mol^-1) and three N application treatments (low-150, normal-250 and high-350 kg N·hm^-2) were included. Soil samples (0-10 cm) were collected for analysis of β-glucosidase, invertase, urease, acid phosphates and β-glucosaminidase activities. The results revealed that with elevated atmospheric CO₂ β-glucosidase activity significantly decreased (P < 0.05) at low N application rates; had no significant effect with a normal N application rate; and significantly increased (P < 0.05) with a high N application rate. For urease activity, at low and normal N application rates (but not high N application rate), elevated atmospheric CO₂ significantly increased (P < 0.05) it. With acid phosphatase elevated atmospheric CO₂ only had significant higher effects (P < 0.05) at high N application rates. Under different CO₂ concentration, effects of N fertilization are also different. Soil β-glucosidase activity at ambient CO₂ concentration decreased with N fertilization, while it increased at elevated CO₂ concentration. In addition, invertase and acid phosphatase activities at elevated CO₂ concentration, significantly increased (P < 0.05) with N treatments, but there was no effect with the ambient CO₂ concentration. For urease activity, at ambient CO₂ concentration, N fertilization increased it significantly (P < 0.05), whereas at elevated CO₂ concentration it was not significant. Additionally, with β-glucosaminidase activity, there were no significant effects from N application. In general, then, elevated atmospheric CO₂ increased soil enzyme activity, which may be attributed to the following two factors: (1) elevated atmospheric CO₂ led to more plant biomass in the soil, which in turn stimulated soil microbial biomass and activity; and (2) elevated atmospheric CO₂ increased plant photosynthesis, thereby increasing plant-derived soil enzymes.     

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in GI pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used mAe. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.     

Global,regional, and cladistic patterns of variation in climatic niche breadths in terrestrial elapid snakes

We obtai ned geo-refere need occurre nee and climatic data from in dividual localities for 59 species of terrestrial elapid snakes, used phylogenetic generalized least squares regression to investigate spatial and cladistic patter ns of variation in climatic niche breadths, and compared patter ns within and across regions and clades to see if they parallel or differ from each other. Specifically, we test (1) whether a species' climatic niche breadth on a given niche axis relates to its position along that axis, and to its climatic niche breadth on another niche axis, and (2) whether variation in niche breadths among species is explained by within-locality variation in climatic conditions or by among-locality variation. We found that:(1) there is an overall global patter n, and patter ns in individual regi ons or clades gen erally parallel each other and global patter ns;(2) species in wanner environments have narrower temperature niche breadths (TNBs);(3) precipitation niche breadth (PNB) and position are positively related;⑷ TNB and PNB are not related;and ⑸ within-locality variation in climatic conditions explains most variation in TNBs, whereas among-locality variation explains most variation in PNBs. Our results are consistent with those reported for lizards of the families Phrynosomatidae and Varanidae, con firm the importance of withi n-locality n iche breadth to species n iche breadth, and show a more im porta nt role of amon g-locality n iche breadth in affecting species niche breadth in terrestrial elapids than in lizards.