Evaluation of Rhodamine 123 As A Probe For Monitoring Mitochondrial Function In Trypanosoma Brucei Spp.

ABSTRACT. Rhodamine 123, a membrane potential-specific dye, has been evaluated as a probe to monitor the function of the mitochondrion in long slender bloodstream and procyclic trypomastigotes of several Trypanosoma brucei spp. By epifluorescence microscopy, mitochondrial development has been followed in long slender bloodstream and procyclic organisms stained with rhodamine 123. to photograph stained long slender bloodstream forms, it was necessary to develop a method to completely immobilize viable organisms. In both parasite forms, as the cell cycle progressed, the mitochondrion developed from a thread-like structure to a highly branched organelle. A dramatic reorganization occurred preceding cytokinesis to produce two progeny thread-like structures which were partitioned into newly formed daughter cells. the organelle within the long slender trypomastigote was found to stain optimally at 0.3 μ/ml of rhodamine 123, while the procyclic form required 3.0 μ/ml. the results suggest that the plasma membrane potential is higher in the long slender parasite than in the procyclic form. the effects of inhibitors that disrupt mitochondrial function were examined in long slender and procyclic parasites, and some of these agents were shown to affect rhodamine 123 accumulation and retention. In long slender trypomastigotes the trypanosome alternative oxidase does not appear to be coupled to proton pumping, whereas in procyclic organisms the effects of inhibitors indicate that this oxidase may be coupled to a pathway that is branched preceding an antimycin A₁-sensitive site.     

枯草芽孢杆菌聚谷氨酸合成途径相关基因功能研究

聚谷氨酸(polyglutamic acid, PGA)作为一种天然多功能的聚合物,近年来成为研究的热点。由于很难通过化学方法合成,微生物发酵是目前生产聚谷氨酸的有效途径。【目的】从基因水平探究枯草芽孢杆菌聚谷氨酸合成途径中degS、degQ、degU、swrA、rocA、putM基因的功能,通过分子改造实现对代谢途径的调控。【方法】以枯草芽孢杆菌为出发菌株,通过对代谢途径中相关基因进行敲除或过表达,分别构建degS、degQ和degU基因缺失的重组菌,swrA、rocA和putM基因过表达的重组菌,借助菌株胞外聚谷氨酸积累的变化分析影响途径的关键节点。【结果】在摇瓶发酵条件下,重组菌Bacillus subtilis 168-swrA、Bacillus subtilis 168-rocA、Bacillus subtilis 168-putM的胞外聚谷氨酸含量分别是原始菌株的1.28倍、1.47倍和1.37倍。重组菌Bacillus subtilis 168-ΔdegS、Bacillus subtilis 168-ΔdegQ、Bacillus subtilis 168-ΔdegU的胞外...     

Trypanosoma Manulis N. Sp. From the Russian Pallas Cat Felis Manul

ABSTRACT. The morphology of Trypanosoma manulis n. sp. is described from living and stained specimens obtained from the blood of a Pallas cat, Felis manul , from Kazakhstan. the cat was also infected with a Hepatozoon sp. and feline immunodeficiency virus. the morphology of the trypanosome most closely resembles that of Trypanosoma mpapuense Reichenow and Trypanosoma heybergi Rodhain found in bats. Trypanosoma manulis does not grow well in conventional media, but co-culture with African green monkey kidney cells in Eagle's Minimum Essential Medium supplemented with 10% fetal calf serum at approximately 27° C resulted in luxuriant growth of trypanosomes. Under these growth conditions, epimastigotes adhered to the surface of the culture flask and to African green monkey kidney cells, as well as forming large rosettes. At 37° C, although growth was poor, transformation of the epimastigotes into the bloodstream forms occurred. This represents the first report of a trypanosome of the subgenus Megatrypanum in a felid.     

Eco-evolution from deep time to contemporary dynamics: The role of timescales and rate modulators

Eco-evolutionary dynamics, or eco-evolution for short, are often thought to involve rapid demography (ecology) and equally rapid heritable phenotypic changes (evolution) leading to novel, emergent system behaviours. We argue that this focus on contemporary dynamics is too narrow: Eco-evolution should be extended, first, beyond pure demography to include all environmental dimensions and, second, to include slow eco-evolution which unfolds over thousands or millions of years. This extension allows us to conceptualise biological systems as occupying a two-dimensional time space along axes that capture the speed of ecology and evolution. Using Hutchinson's analogy: Time is the ‘theatre’ in which ecology and evolution are two interacting ‘players’. Eco-evolutionary systems are therefore dynamic: We identify modulators of ecological and evolutionary rates, like temperature or sensitivity to mutation, which can change the speed of ecology and evolution, and hence impact eco-evolution. Environmental change may synchronise the speed of ecology and evolution via these rate modulators, increasing the occurrence of eco-evolution and emergent system behaviours. This represents substantial challenges for prediction, especially in the context of global change. Our perspective attempts to integrate ecology and evolution across disciplines, from gene-regulatory networks to geomorphology and across timescales, from today to deep time.     

生态系统稳态转换检测研究进展

在气候变化、人类活动等影响下,生态系统结构和功能可能发生大规模的突变,导致生态系统从一个相对稳定的状态进入另一个稳定状态,这种现象称为稳态转换。由于生态系统的复杂性,准确刻画生态系统多稳态并界定其临界点尚存在挑战,提升对生态系统稳态转换的检测和预测能力依旧是生态学领域研究的热点和难题。基于多稳态理论和稳态转换经典概念框架,阐释了稳态转换检测的理论基础;归纳总结出四种稳态转换检测方法的原理和优劣势;鉴于稳态转换的尺度依赖性,梳理了单一生态系统、区域综合生态系统和全球生态系统不同尺度下的稳态转换检测方法、研究思路和应用案例。基于研究进展和问题现状,提出在未来研究中,亟待发展适应复杂系统的综合检测方法;创新稳态转换多尺度分析的技术方法体系;深化生态系统稳态转换驱动机制研究,构建多元耦合机理模型;进而深化稳态转换检测结果链接生态系统管理的实践研究;解析生态系统服务和可持续发展机制。     

基于CRISPR/Cas系统的多重基因编辑与调控技术

规律成簇的间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)及其相关Cas蛋白所构建的CRISPR/Cas系统是古细菌或细菌中特有的一种获得性免疫系统。研究人员将其开发成基因编辑工具之后,凭借其高效、精准和通用性强等优点迅速成为合成生物学领域的热门研究方向,在生命科学、生物工程技术、食品科学及农作物育种等多个领域引发了革命性的影响。目前基于CRISPR/Cas系统单基因编辑与调控技术日益完善,但在多重基因编辑和调控方面仍存在挑战。本文聚焦基于CRISPR/Cas系统的多重基因编辑与调控技术开发及应用,针对单个细胞内实现多位点基因编辑或调控和细胞群体内实现多位点基因编辑或调控技术,依据作用原理对其进行了系统总结和阐述,包括基于CRISPR/Cas系统的双链断裂、单链断裂以及多重基因调控技术等。这些工作丰富了多重基因编辑与调控的工具,为CRISPR/Cas系统在多领域的应用作出了贡献。     

拟南芥ACOL8基因在乙烯合成与响应中的功能分析

植物激素乙烯在多种生理生化过程中发挥重要作用,但其在特定组织器官中的合成机制尚不完全清楚。拟南芥中存在12个功能未知的ACC氧化酶类似蛋白（ACO-like homolog,ACOL）,运用基因定点编辑技术构建了ACOL8的功能丧失型突变体,发现该基因的突变削弱了经典的乙烯“三重反应”。与野生型相比,突变体黄化幼苗下胚轴及主根的长度显著增加,这与突变体对外源ACC的敏感性下降现象一致。同时还发现ACOL8基因的表达受乙烯信号的正反馈调控,EIN3过表达增强其表达水平,而etr1-3的突变则产生相反效应。再者,在正常条件下,ACOL8基因的突变并未影响拟南芥的生长;但在盐胁迫条件下,突变体的根冠比显著下降,这说明该基因参与植物的盐胁迫响应。综上,这些结果说明ACOL8可能具有ACC氧化酶的功能,参与乙烯的合成与响应。     

The application of reproductive technology to endangered species breeding programmes

Captive breeding plays an increasingly important role in species conservation, but special problems are encountered in achieving the ideal of a demographically stable but genetically diverse population. Breeding programmes involving co-operation among a number of centres are now being developed which will overcome some of these difficulties by identifying individual animals, genetic lineages or age cohorts from which to breed. Application of techniques such as artificial insemination, embryo transfer and semen collection and storage, as well as the monitoring of reproductive status will contribute to the success of such programmes. The usefulness of these procedures for various population problems is discussed and criteria for their appropriate implementation within breeding programmes is outlined.