Sorting nexin Snx41 is essential for conidiation and mediates glutathione-based antioxidant defense during invasive growth in Magnaporthe oryzae

The sorting nexins Atg20/Snx42 and Snx41 regulate membrane traffic and endosomal protein sorting and are essential for Cvt and/or pexophagy in yeast. Previously, we showed that macroautophagy is necessary for conidiation in the rice-blast fungus Magnaporthe oryzae. Here, we analyzed the physiological function(s) of selective autophagy in Magnaporthe through targeted deletion of MGG_12832, an ortholog of yeast SNX41 and ATG20/SNX42. Loss of MGG_12832 (hereafter SNX41) abolished conidia formation and pathogenesis in M. oryzae. Snx41-GFP localized as dynamic puncta or short tubules that are partially associated with autophagosomes and/or autophagic vacuoles. PX domain, but not macroautophagy per se, was required for such localization of Snx41-GFP in Magnaporthe. Although not required for nonselective autophagy, Snx41 was essential for pexophagy in Magnaporthe. We identified Oxp1, an ATP-dependent oxoprolinase in the gamma-glutamyl cycle, as a binding partner and potential retrieval target of Snx41-dependent protein sorting. The substrate of Oxp1, 5-oxoproline, could partially restore conidiation in the snx41Δ. Exogenous glutathione, a product of the gamma-glutamyl cycle, significantly restored pathogenicity in the snx41Δ mutant, likely through counteracting the oxidative stress imposed by the host. We propose that the gamma-glutamyl cycle and glutathione biosynthesis are subject to regulation by Snx41-dependent vesicular trafficking, and mediate antioxidant defense crucial for in planta growth and pathogenic differentiation of Magnaporthe at the onset of blast disease in rice.     

Sorting nexin Snx41 is essential for conidiation and mediates glutathione-based antioxidant defense during invasive growth in Magnaporthe oryzae

The sorting nexins Atg20/Snx42 and Snx41 regulate membrane traffic and endosomal protein sorting and are essential for Cvt and/or pexophagy in yeast. Previously, we showed that macroautophagy is necessary for conidiation in the rice-blast fungus Magnaporthe oryzae. Here, we analyzed the physiological function(s) of selective autophagy in Magnaporthe through targeted deletion of MGG_12832, an ortholog of yeast SNX41 and ATG20/SNX42. Loss of MGG_12832 (hereafter SNX41) abolished conidia formation and pathogenesis in M. oryzae. Snx41-GFP localized as dynamic puncta or short tubules that are partially associated with autophagosomes and/or autophagic vacuoles. PX domain, but not macroautophagy per se, was required for such localization of Snx41-GFP in Magnaporthe. Although not required for nonselective autophagy, Snx41 was essential for pexophagy in Magnaporthe. We identified Oxp1, an ATP-dependent oxoprolinase in the gamma-glutamyl cycle, as a binding partner and potential retrieval target of Snx41-dependent protein sorting. The substrate of Oxp1, 5-oxoproline, could partially restore conidiation in the snx41Δ. Exogenous glutathione, a product of the gamma-glutamyl cycle, significantly restored pathogenicity in the snx41Δ mutant, likely through counteracting the oxidative stress imposed by the host. We propose that the gamma-glutamyl cycle and glutathione biosynthesis are subject to regulation by Snx41-dependent vesicular trafficking, and mediate antioxidant defense crucial for in planta growth and pathogenic differentiation of Magnaporthe at the onset of blast disease in rice.     

Pexophagy: The Selective Autophagy of Peroxisomes

Pichia pastoris and Hanseula polymorpha are methylotrophic yeasts capable of utilizing methanol, as a sole source of carbon and energy. Growth of these yeast species on methanol requires the synthesis of cytosolic and peroxisomal enzymes combined with the proliferation of peroxisomes. Peroxisomes are also abundantly present in the alkane-utilizing yeast Yarrowia lipolytica upon growth of cells on oleic acid. This feature has made these yeast species attractive model systems to dissect the molecular mechanisms controlling peroxisome biogenesis. We have found that upon glucose- or ethanol-induced catabolite inactivation of metabolically superfluous peroxisomes are rapidly and selectively degraded within the vacuole by a process called pexophagy, the selective removal of peroxisomes by autophagy-like processes. Utilizing several genetic screens, we have identified a number of genes that are essential for pexophagy. In this review, we will summarize our current knowledge of the molecular events of pexophagy.     

钙在植物花发育过程中的作用

对于园林观赏植物,开花是一个非常重要的发育阶段,它直接影响花卉的品质。近年来,植物花发育的分子生物学研究进展迅速,并取得了一些突破性成果。钙作为第二信使在植物信号转导中起着非常重要的作用,大量研究显示,钙有可能参与开花控制。本文总结了钙信号与植物花发育这一领域的最新研究进展,包括以下几个方面的内容：钙在植物成花诱导(包括光周期诱导和低温诱导)中的作用;花芽分化时期钙在植物叶芽和花芽中的动态分布及组织培养条件下不同钙浓度对花芽分化的影响;钙与花衰老的关系。     

JMJ在胚胎发育中的作用

胚胎发育是一个连续、复杂的过程,涉及到多种基因的参与。JMJ是广泛存在的转录抑制因子,包括核定位信号区、转录抑制区、DNA结合区,是富含A/T相互作用因子家族中的一员。它通过与一些转录调控蛋白的相互作用调节细胞生长和基因转录,在胚胎器官形态形成中发挥重要的作用。     

NEDD化修饰系统及其在肿瘤发展中的作用

目前已经鉴定出17种类泛素蛋白（ubiquitin like proteins,UBLs）,这些蛋白与底物的结合方式与泛素相似．根据进化特征,可将UBLs分为9类,分别为：NEDD8、SUMO、ISG15、FUB1、FAT10、Atg8、Atg12、Urm1和UFM1．NEDD8是目前研究最多的UBLs之一,与泛素的氨基酸序列具有高度相似性．NEDD化修饰是一种动态的可逆蛋白质翻译后修饰方式,可以将NEDD8共价结合到靶蛋白之上,也可以将NEDD8从靶蛋白上去除．NEDD化修饰对蛋白功能具有重要的调节作用,如改变蛋白质的空间构象、阻碍底物与其它蛋白质的相互作用和招募与NEDD8相互作用的蛋白等．最新研究表明,NEDD化与肿瘤的发生发展密切相关,但具体的机制还不清楚．本文将就NEDD化修饰在肿瘤发展过程中的作用机制做一综述．     

钙在植物花发育过程中的作用

对于园林观赏植物,开花是一个非常重要的发育阶段,它直接影响花卉的品质。近年 来,植物花发育的分子生物学研究进展迅速,并取得了一些突破性成果。钙作为第二信使在植物信号转导中起着非常重要的作用,大量研究显示,钙有可能参与开花控制。本文总结了钙信号与植物花发育这一领域的最新研究进展,包括以下几个方面的内容：钙在植物成花诱 导（包括光周期诱导和低温诱导）中的作用;花芽分化时期钙在植物叶芽和花芽中的动态分布及组织培养条件下不同钙浓度对花芽分化的影响;钙与花衰老的关系。      

Role of lysine residues of the Magnaporthe oryzae effector AvrPiz-t in effector- and PAMP-triggered immunity

Magnaporthe oryzae is an important fungal pathogen of both rice and wheat. However, how M. oryzae effectors modulate plant immunity is not fully understood. Previous studies have shown that the M. oryzae effector AvrPiz-t targets the host ubiquitin-proteasome system to manipulate plant defence. In return, two rice ubiquitin E3 ligases, APIP6 and APIP10, ubiquitinate AvrPiz-t for degradation. To determine how lysine residues contribute to the stability and function of AvrPiz-t, we generated double (K1,2R-AvrPiz-t), triple (K1,2,3R-AvrPiz-t) and lysine-free (LF-AvrPiz-t) mutants by mutating lysines into arginines in AvrPiz-t. LF-AvrPiz-t showed the highest protein accumulation when transiently expressed in rice protoplasts. When co-expressed with APIP10 in Nicotiana benthamiana, LF-AvrPiz-t was more stable than AvrPiz-t and was less able to degrade APIP10. The avirulence of LF-AvrPiz-t on Piz-t:HA plants was less than that of AvrPiz-t, which led to resistance reduction and lower accumulation of the Piz-t:HA protein after inoculation with the LF-AvrPiz-t-carrying isolate. Chitin- and flg22-induced production of reactive oxygen species (ROS) was higher in LF-AvrPiz-t than in AvrPiz-t transgenic plants. In addition, LF-AvrPiz-t transgenic plants were less susceptible than AvrPiz-t transgenic plants to a virulent isolate. Furthermore, both AvrPiz-t and LF-AvrPiz-t interacted with OsRac1, but the suppression of OsRac1-mediated ROS generation by LF-AvrPiz-t was significantly lower than that by AvrPiz-t. Together, these results suggest that the lysine residues of AvrPiz-t are required for its avirulence and virulence functions in rice.     

Structure-Function Analysis of Rgs1 in Magnaporthe oryzae: Role of DEP Domains in Subcellular Targeting

Background

Rgs1, a prototypical Regulator of G protein Signaling, negatively modulates the cyclic AMP pathway thereby influencing various aspects of asexual development and pathogenesis in the rice-blast fungus Magnaporthe oryzae. Rgs1 possesses tandem DEP motifs (termed DEP-A and DEP-B; for Dishevelled, Egl-10, Pleckstrin) at the N-terminus, and a Gα-GTP interacting RGS catalytic core domain at the C-terminus. In this study, we focused on gaining further insights into the mechanisms of Rgs1 regulation and subcellular localization by characterizing the role(s) of the individual domains and the full-length protein during asexual development and pathogenesis in Magnaporthe.

Methodology/Principal Findings

Utilizing western blot analysis and specific antisera against the N- and C-terminal halves of Rgs1, we identify and report the in vivo endoproteolytic processing/cleavage of full-length Rgs1 that yields an N-terminal DEP and a RGS core domain. Independent expression of the resultant DEP-DEP half (N-Rgs1) or RGS core (C-Rgs1) fragments, failed to complement the rgs1Δ defects in colony morphology, aerial hyphal growth, surface hydrophobicity, conidiation, appressorium formation and infection. Interestingly, the full-length Rgs1-mCherry, as well as the tagged N-terminal DEP domains (individually or in conjunction) localized to distinct punctate vesicular structures in the cytosol, while the catalytic RGS core motif was predominantly vacuolar.

Conclusions/Significance

Based on our data from sequence alignments, immuno-blot and microscopic analysis, we propose that the post-translational proteolytic processing of Rgs1 and the vacuolar sequestration of the catalytic RGS domain represents an important means of down regulating Rgs1 function and thus forming an additional and alternative means of regulating G protein signaling in Magnaporthe. We further hypothesize the prevalence of analogous mechanisms functioning in other filamentous fungi. Furthermore, we conclusively assign a specific vesicular/membrane targeting function for the N-terminal DEP domains of Rgs1 in the rice-blast fungus.     

The Role of SETAC in the Development of LCA

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DOI: http://dx.doi.org/10.1065/lca2006.04.019

Background

Life cycle assessments have been performed using different methods before the name was coined since about 1970 in several countries of North America and Europe. It was the merit of SETAC to start a standardization process which culminated in the LCA-guidelines ('A code of practice') in 1993. It is the aim of this paper to trace back this and further LCA-related achievements by SETAC on the basis of documents and personal memories. It may be subjective in the selection and weighting of some events, but objectivity is strived for with regard to the whole and, in my view, singular development.

Results and Discussion

Starting 1990 with two workshops in Smuggler's Notch (Vermont) and Leuven (Belgium), SETAC and SETAC Europe organized several workshops during which important topics (framework, impact assessment, data quality, etc.) were treated and published in the form of reports which are still available. The main contribution by CML and its head, Helias Udo de Haes, was a practical method of impact assessment, transforming the formerly more technocratic LCA (energy, resources, waste) into an instrument of environmental assessment of product systems. In addition, important contributions to the allocation problem were made. Starting in 1993, ISO took over the leadership in standardization and SETAC started the famous working groups in North America and Europe, often dealing with the same topics in parallel. Due to the different cultures, the results were frequently complimentary rather than harmonic. The CML-method of LCIA, widely accepted in Europe, had to wait for about 10 years to be accepted at the other side of the Atlantic. It was helpful that SETAC – meanwhile a global organization – looked for a partner in order to implement LCA all over the world. This partner was found in the 'United Nations Environmental Programme' (UNEP) and the UNEP/SETAC Life Cycle Initiative was officially launched by Klaus Töpfer in Prague in April 2002. SETAC also assumed an important role in communicating LCA via publications: workshop and conference reports, the 'code of practice', working group results and LCA News Letters. The annual meetings offered forums for LCA scientists, practitioners and users, well prepared by the LCA Steering Committee (SETAC Europe) and the LCA Advisory Group (SETAC North America).

Recommendation

. The main recommendation to SETAC is to adhere to LCA as the main environmental assessment tool for products and to expand it to a true sustainability assessment tool by adding Life Cycle Costing (LCC) and a still to be invented 'Social Life Cycle Assessment'. SETAC is to remain the scientific arm within the UNEP/SETAC LC Initiative, without loosing its identity. Working groups should be global rather than regional in the future, as suggested by the SETAC Europe LCA Steering Committee at the 2004 World Congress in Portland, Oregon.

     

The Role of Cooperation in the Development of Intelligence

In the theoretical analysis of the general structure of activity as presented in the fundamental work of L. S. Vygotsky and A. N. Leont'ev, cooperation among the participants in a joint activity is viewed as the source of new mental functions. Under conditions of cooperation, an activity that is initially shared by those participating in it emerges as an original and fundamental foundation for the development of forms of individual activity. Thus, L. S. Vygotsky wrote, "Any function in the cultural development of the child appears twice, on two levels: first as a social function, and then as a psychological function, first as something taking place between people, a category based on relationship between minds, an intermental category; later, this psychological function occurs within the child, as an intramental category" [2. Pp. 197-98].     

瘦素在脑老化认知发展不同阶段中的作用

脑老化过程中伴随脑结构、功能退化和认知能力减退,其认知发展呈异质性,即发展的不均一性,有成功脑老化、正常脑老化、轻度认知功能障碍和阿尔茨海默病等状态。瘦素在不同认知状态下含量不同。研究发现,无神经病理改变的老年人群瘦素水平高,认知功能减退不明显;多数轻度认知功能障碍者瘦素水平降低;瘦素可改善阿尔茨海默病患者的认知障碍,被认为是一种潜在的认知增强剂。瘦素水平降低在脑老化认知障碍的发展中起重要作用。     

膨胀素在植物生长发育中的作用

膨胀素是一类具有非水解活性的细胞壁松弛蛋白,参与植物生长发育过程中细胞壁的修饰。大多数植物中都发现有膨胀素基因家族成员存在,其功能涉及植物生长发育的各个方面,包括营养生长、形态发生、受精授粉、果实成熟等,并表现出高度的组织、器官和细胞特异性。本文综述膨胀素在种子萌发、叶的发育、根茎生长、花的发育等生长发育过程中的作用。     

The Role of Autophagy During Development in Higher Eukaryotes

Autophagy is a lysosome‐mediated degradation pathway used by eukaryotes to recycle cytosolic components in both basal and stress conditions. Several genes have been described as regulators of autophagy, many of them being evolutionarily conserved from yeast to mammals. The study of autophagy‐defective model systems has made it possible to highlight the importance of correctly functioning autophagic machinery in the development of invertebrates as, for example, during the complex events of fly and worm metamorphosis. In vertebrates, on the other hand, autophagy defects can be lethal for the animal if the mutated gene is involved in the early stages of development, or can lead to severe phenotypes if the mutation affects later stages. However, in both lower and higher eukaryotes, autophagy seems to be crucial during embryogenesis by acting in tissue remodeling in parallel with apoptosis. An increase of autophagic cells is, in fact, observed in the embryonic stages characterized by massive cell elimination. Moreover, autophagic processes probably protect cells during metabolic stress and nutrient paucity that occur during tissue remodeling. In light of such evidence, it can be concluded that there is a close interplay between autophagy and the processes of cell death, proliferation and differentiation that determine the development of higher eukaryotes.     

类脂对植物生长和发育的作用

简述了类脂在植物生长和发育中的作用,特别是类脂中脂肪酸的饱和度对植物生长发育的影响,植物固醇对植物的表型和在低温下的生长、胚胎的发育以及可育性中的作用.