Neonothopanus gardneri: a new combination for a bioluminescent agaric from Brazil

The bioluminescent agaric, Agaricus gardneri Berk., was rediscovered recently in central Brazil. The new combination, Neonothopanus gardneri, is proposed for this long-forgotten taxon supported by morphological and molecular data.     

宽阔水自然保护区的虫草及其相关真菌Ⅰ.

本文报道采自贵州省宽阔水自然保护区的9种虫草,它们是多壳虫草新种(Cordyceps polycarpica Liang & Liu Sp.nov.),鼠尾虫草新种(Cordyceps musicaudata Liang et Liu sp.nov.),针孢虫草(Cordyceps acicularis Rev.ex Berk.),罗伯茨虫草(Cordyceps robertsii(Hook.)Gray),下垂虫草(Cordyceps nutans Pat.),沫蝉虫草(Cordyceps tricentri Yasuda),丝虫草(Cordyceps filiformis Moureau),泽地虫草(Cordyceps paludosa Mains)和球头虫草[Cordyceps sphecocephala(K1.)Sacc.]。     

The delimitation of Gelasine (Iridaceae), and G. uruguaiensis sp. nov. from Uruguay

The generic concept of Gelasine Herb. (Iridaceae) is reviewed, and the characters discussed in connection with Sphenostigma Bak., a synonym of Gelasine. The reduction of Sphenostigma is further justified on account of overlapping features. A new species from Uruguay, G. uruguaiensis Rav. sp. nov., is described and illustrated. This is shown to include two subspecies: ssp. uruguaiensis , and ssp. orientalis ssp. nov. The chromosome number of the latter is 2n = 14.     

中国韧革菌(Ⅲ)

本文报道中国韧革菌科(Stereaceae Pilat)真菌17种,隶属于柄革菌属(Stereopsis Reid)。韧革菌属(Stereum Hill:Persoon)和刷革菌属(Xylobolus Karsten)。其中有3个新种,1个新组合种和2个新记录种,它们是:厚盖柄革菌(Stereopsis crassipileata Z.T.Guo,sp.nov.)、细柄柄革菌(S.gracilistipitata Z.T.Guo,sp.nov.),假杯柄革菌(S.pseudocupulata Z.T.Guo,sp。nov.)、掌状柄革菌(S.craspedia(Fr.)Z.T.Guo,comb.nov.)、瓣裂柄革菌(S.hiscens(Berk. & Rav.)Reid)和蛋黄柄革菌(S.uitellina(Plowr.)Reid)。标本全部保藏于中国科学院微生物研究所真菌标本室(HMAS)。     

SYNAPTONEMAL COMPLEXES IN RED ALGAE1,2

The presence of synaptonemal complexes in the nuclei of young tetraspore mother cells is described for the first time in the red algae. Synaptonemal complexes were found in Janczewskia gardneri Setchell, Levringiella gardneri Setchell, Gonimophyllum skottsbergii Setchell, and Polycoryne gardneri Setchell. The synaptonemal complexes consist of 2 lateral, dark-staining elements from which small fibrils extend to form a less densely staining central element. With minor variations, the dimensions and structure of these synaptonemal complexes correspond to those found in other organisms.     

Molecular Interactions and Cellular Itinerary of the Yeast RAVE (Regulator of the H+-ATPase of Vacuolar and Endosomal Membranes) Complex

The RAVE complex (regulator of the H⁺-ATPase of vacuolar and endosomal membranes) is required for biosynthetic assembly and glucose-stimulated reassembly of the yeast vacuolar H⁺-ATPase (V-ATPase). Yeast RAVE contains three subunits: Rav1, Rav2, and Skp1. Rav1 is the largest subunit, and it binds Rav2 and Skp1 of RAVE; the E, G, and C subunits of the V-ATPase peripheral V₁ sector; and Vph1 of the membrane V_o sector. We identified Rav1 regions required for interaction with its binding partners through deletion analysis, co-immunoprecipitation, two-hybrid assay, and pulldown assays with expressed proteins. We find that Skp1 binding requires sequences near the C terminus of Rav1, V₁ subunits E and C bind to a conserved region in the C-terminal half of Rav1, and the cytosolic domain of Vph1 binds near the junction of the Rav1 N- and C-terminal halves. In contrast, Rav2 binds to the N-terminal domain of Rav1, which can be modeled as a double β-propeller. Only the V₁ C subunit binds to both Rav1 and Rav2. Using GFP-tagged RAVE subunits in vivo, we demonstrate glucose-dependent association of RAVE with the vacuolar membrane, consistent with its role in glucose-dependent V-ATPase assembly. It is known that V₁ subunit C localizes to the V₁-V_o interface in assembled V-ATPase complexes and is important in regulated disassembly of V-ATPases. We propose that RAVE cycles between cytosol and vacuolar membrane in a glucose-dependent manner, positioning V₁ and V₀ subcomplexes and orienting the V₁ C subunit to promote assembly.     

Skp1p regulates Soi3p/Rav1p association with endosomal membranes but is not required for vacuolar ATPase assembly

Skp1p is an essential component of SCF-type E3 ubiquitin ligase complexes and associates with these through binding to F-box proteins. Skp1p also binds F-box proteins in a number of non-SCF complexes. The Skp1p-associated yeast protein Soi3p/Rav1p (hereafter referred to as Rav1p) is a component of the RAVE complex required for regulated assembly of vacuolar ATPase (V-ATPase). Rav1p is also involved in transport of TGN proteins and endocytic cargo between early and late endosomes. To evaluate the role of Skp1p in the RAVE complex, we made use of the fact that overexpression of Rav1p is toxic because it sequesters Skp1p from essential interactions. We isolated a separation of function allele of SKP1, skp1(Asn108Tyr), that completely abrogated the Rav1p interaction but allowed Skp1p to perform other essential cellular functions. Cells containing the skp1(Asn108Tyr) allele as the sole source of Skp1p exhibited normal V-ATPase assembly and activity. However, in the skp1(Asn108Tyr) mutant strain, the membrane-associated pool of Rav1-green fluorescent protein was increased, suggesting that Skp1p is important for the release of Rav1p from endosomal membranes where it functions in V-ATPase assembly. Thus, although part of the RAVE complex, Skp1p does not appear to be involved in V-ATPase assembly but instead in the cycling of the complex off membranes. This work also provides a generalizable approach to defining the roles of interactions of Skp1p with individual F-box proteins through the isolation of special alleles of SKP1.     

Development of microsatellite markers for the moss Ptychomitrium gardneri (Ptychomitriaceae)

? Premise of the study: Microsatellite markers were developed for Ptychomitrium gardneri to study population genetics of this eastern Asian-North American disjunct moss. ? Methods and Results: A total of 13 microsatellite markers were developed in Chinese populations of P. gardneri, using the Fast Isolation by AFLP of Sequences COntaining Repeats protocol. Eight of the markers showed polymorphism when assessed in a sample of four populations of 29 individuals from China. These markers amplified three to four alleles per locus. Five primers also amplified in P. linearifolium and P. wilsonii. ? Conclusions: These markers may be useful for further investigation of population genetics of P. gardneri.     

Canker and wilt of black locust (Robinia pseudoacacia L.) caused by Fusarium species

From the pathological material of black locust trees showing symptoms of wilting of the foliage or canker of the bark the following Fusarium species were isolated: Fusarium avenaceum (Fr.) Sacc., Fusarium lateritium Nees., Fusarium semitectum Berk. & Rav., Fusarium solani (Mart.) Sacc., Fusarium sulphureum Schlecht. (syn.: Fusarium sambucinum Fuckel f. 6 Wollenw.) The results of the provocation infections of one-year-old black locust seedlings showed that all of the species--except Fusarium solani--are able to cause considerable necrosis in living bark and phloem. Fusarium sulphureum had by far the highest pathogenecity among the tested species. Fusarium semitectum isolated from withered black locust tree also caused necrosis on significant bark area. In the course of the penetration assay Fusarium sulphureum and Fusarium avenaceum were the most successful, and these species can cause cankers on the stem and twigs of black locust without frost effect.     

Myristicaceae novelties from Ecuador

Virola aequatorialis is described as a new species from the coastal plain of Ecuador; it differs from the southeast Brazilian V. gardneri in several characters including its prominently reticulate leaves and the anthers which are obtuse and slightly shorter than the filament column. Osteophloeum platyspermum var. sulcatum and Otoba glycycarpa are proposed as new combinations.     

Skp1 forms multiple protein complexes, including RAVE, a regulator of V-ATPase assembly

SCF ubiquitin ligases are composed of Skp1, Cdc53, Hrt1 and one member of a large family of substrate receptors known as F-box proteins (FBPs). Here we report the identification, using sequential rounds of epitope tagging, affinity purification and mass spectrometry, of 16 Skp1 and Cdc53-associated proteins in budding yeast, including all components of SCF, 9 FBPs, Yjr033 (Rav1) and Ydr202 (Rav2). Rav1, Rav2 and Skp1 form a complex that we have named 'regulator of the (H+)-ATPase of the vacuolar and endosomal membranes' (RAVE), which associates with the V1 domain of the vacuolar membrane (H+)-ATPase (V-ATPase). V-ATPases are conserved throughout eukaryotes, and have been implicated in tumour metastasis and multidrug resistance, and here we show that RAVE promotes glucose-triggered assembly of the V-ATPase holoenzyme. Previous systematic genome-wide two-hybrid screens yielded 17 proteins that interact with Skp1 and Cdc53, only 3 of which overlap with those reported here. Thus, our results provide a distinct view of the interactions that link proteins into a comprehensive cellular network.     

Lijndenia, a re–established paleotropical genus of the Melastomataceae–Memecyleae

The genus Lijndenia Zoll. & Mor. is re–established and emended to include four species, L. laurina Zoll. & Mor. from W Malesia and Java, L. capitellata (Arn.) Bremer from Ceylon, L. gardneri (Thw.) Bremer from Ceylon, and L. barteri (Hook. f.) Bremer from tropical W Africa. The latter three names are new combinations.     

Confirmation of cross-fertilization using molecular markers in ornamental passion flower hybrids

Several interspecific Passiflora hybrids are produced in the northern hemisphere for the ornamental plant market. In Brazil, production of passion flower hybrids is limited to the introgression of genes into the main cultivated species, yellow passion fruit, to be used as rootstocks. Confirmation of hybridization in the initial developmental stage is important for breeding perennial and sub-perennial plants, such as passion flowers, reducing time and costs in plant stock maintenance. In order to obtain F? hybrids with ornamental potential, four species of Passiflora (P. alata, P. gardneri, P. gibertii, and P. watsoniana) from the Active Germplasm Bank at UESC were hybridized. Flower buds, in pre-anthesis, of the genitors were previously protected, and the female buds were emasculated. To confirm hybridization, the genomic DNA of the genitor species and the supposed hybrids was extracted and RAPD primers were used to obtain molecular markers and select passion flower interspecific hybrids. Eight primers were used to confirm hybrids derived from P. gardneri with P. alata, P. watsoniana with P. alata, P. watsoniana with P. gardneri, and P. gardneri with P. gibertii; 75, 50, 45, and 46% of the informative bands, respectively, confirmed the hybrid nature of these plants. The RAPD technique was effective in the early identification of hybrids; this will be useful for development of hybrid Passiflora progeny.     

Characterization of the complex involved in regulating V-ATPase activity of the vacuolar and endosomal membrane

Regulator of the H⁺-ATPase of the vacuolar and endosomal membranes (RAVE) is essential for the reversible assembly of H⁺-ATPase. RAVE primarily consists of three subunits: Rav1p, Rav2p and Skp1p. To characterize these subunits, in this study, four strains derived from Saccharomyces cerevisiae BY4742 were constructed with a FLAG tag on the Rav1p and Rav2p subunits. Then, the corresponding RAVE containing complex was isolated by affinity purification. Western blot and MALDI-TOF mass spectrometry analyses showed that the RAVE complex contains not only the known V₁-ATPase subunits (Vma1p and Vma2p) but also a newly found Leu1p that interacts with the RAVE subunit. Furthermore, we constructed rav1?/rav2?/vma2?/leu1-deficient recombinants by fusion PCR and homologous recombination and demonstrated that leu1 is indispensable in adjusting the microbial cell to adverse environments and that the function is similar to that of rav1/rav2 but significantly differs from that of vma2. Leu1p probably plays an important role in RAVE regulation of V-ATPase activity in conjunction with RAVE.     

The genus Iphiona (Compositae-Inuleae)

The genus Iphiona Cass. emend. A. Anderb. (Compositae-Inuleae-Inulinae) is revised and considerably amended. The former genera Grantia Boiss. (= Perralde-riopsis Rauschert) and Hirschia Baker are included as well as the major part of Iphiona Cass. s.str. and the species Inula grantioides Boiss., and Pulicaria phillipsiae S. Moore. Five new combinations are made: I. anthemidifolia (Baker) A. Anderb., I. arachnoidea (Boiss.) A. Anderb., I. grantioides (Boiss.) A. Anderb., I. phillipsiae (S. Moore) A. Anderb., and I. senecionoides (Baker) A. Anderb. The new species I. teretifolia A. Anderb. is described. A discussion of the phylogeny of the group and a cladogram, involving reticulation, of the Iphiona species are presented. The 11 species are distributed in Egypt, Israel, Jordan, Sudan, Ethiopia, Somalia, Saudi Arabia, Peoples Democratic Republic Yemen, Oman, Iran and Pakistan.     

利用蛋白标签纯化酿酒酵母RAVE-V_1复合物

RAVE(regulator of the H+-ATPase of the vacuolar and endosomal membranes)是调节液泡ATP酶(V-ATP酶)装配与拆卸过程的调节酶,由Rav1p、Rav2p和Skp1p 3个亚基构成。在酿酒酵母细胞中,当葡萄糖耗尽时,V-ATP酶分解成V1、V0两部分,此时,RAVE与V1以复合物的形式存在于细胞质中。本研究利用同源重组技术,构建在基因RAV2的3'端定点插入FLAG标签的重组菌株BY4742 RAV2-FLAG,通过亲和层析原理纯化RAVE-V1复合物,为后续利用电子显微镜对其进行三维结构研究奠定坚实的基础。结果表明:FLAG标签添加到Rav2p的C端可以成功纯化出RAVE-V1复合物;结合质谱鉴定首次发现了Leu1p与RAVE存在相互作用关系,这使得对RAVE的研究转向一个全新的方向;此外,本研究方案对其他调节蛋白及与之相互作用的蛋白组的分离纯化具有借鉴意义。     

A multilocus sequence analysis of the genus Xanthomonas

A multilocus sequence analysis (MLSA) of strains representing all validly published Xanthomonas spp. (119 strains) was conducted using four genes; dnaK, fyuA, gyrB and rpoD, a total of 440 sequences. Xanthomonas spp. were divided into two groups similar to those indicated in earlier 16S rDNA comparative analyses, and they possibly represent distinct genera. The analysis clearly differentiated most species that have been established by DNA-DNA reassociation. A similarity matrix of the data indicated clear numerical differences that could form the basis for species differentiation in the future, as an alternative to DNA-DNA reassociation. Some species, X. cynarae, X. gardneri and X. hortorum, formed a single heterogeneous group that is in need of further investigation. X. gardneri appeared to be a synonym of X. cynarae. Recently proposed new species, X. alfalfae, X. citri, X. euvesicatoria, X. fuscans and X. perforans, were not clearly differentiated as species from X. axonopodis, and X. euvesicatoria and X. perforans are very probably synonyms. MLSA offers a powerful tool for further investigation of the classification of Xanthomonas. Based on the dataset produced, the method also offers a relatively simple way of identifying strains as members of known species, or of indicating their status as members of new species.