Appressorium-mediated penetration of Magnaporthe oryzae and Colletotrichum orbiculare into surface-cross-linked agar media

This transmission electron microscopy (TEM) procedure was designed to examine a gram positive spore-forming bacillus in colony on various solid agar media with minimal artifact. Cellular morphology and organization of colonies embedded in Poly/Bed 812 resin (P/B) were studied. It is a modification of procedures used for undecalcified rat bone and Stomatococcus mucilaginosus.Cultures were fixed and processed at room temperature (RT) under a fume hood. Solutions were added with a Pasteur pipet and removed by gentle vacuum aspiration. Other equipment used is shown in Figure 3. Cultures were fixed for 17-18 h in 10-20 ml of RT 2% phosphate buffered glutaraldehyde (422 mosm/KgH2O) within 5 m after removal from the incubator. After 3 (30 m) changes in 0.15 M phosphate buffer (PB = 209-213 mosm/KgH2O, pH 7.39-7.41), colony cut-outs (CCO) were made with a scalpel.

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Crystal structures of Magnaporthe oryzae trehalose-6-phosphate synthase (MoTps1) suggest a model for catalytic process of Tps1

Biochemical Journal ◽

10.1042/bcj20190289 ◽

2019 ◽

Vol 476 (21) ◽

pp. 3227-3240 ◽

Cited By ~ 1

Author(s):

Shanshan Wang ◽

Yanxiang Zhao ◽

Long Yi ◽

Minghe Shen ◽

Chao Wang ◽

...

Keyword(s):

Crystal Structures ◽

Conformational Changes ◽

Magnaporthe Oryzae ◽

Structural Information ◽

Complex Structure ◽

Critical Role ◽

Catalytic Process ◽

Structural Basis ◽

Salt Bridges ◽

Terminal Domain

Trehalose-6-phosphate (T6P) synthase (Tps1) catalyzes the formation of T6P from UDP-glucose (UDPG) (or GDPG, etc.) and glucose-6-phosphate (G6P), and structural basis of this process has not been well studied. MoTps1 (Magnaporthe oryzae Tps1) plays a critical role in carbon and nitrogen metabolism, but its structural information is unknown. Here we present the crystal structures of MoTps1 apo, binary (with UDPG) and ternary (with UDPG/G6P or UDP/T6P) complexes. MoTps1 consists of two modified Rossmann-fold domains and a catalytic center in-between. Unlike Escherichia coli OtsA (EcOtsA, the Tps1 of E. coli), MoTps1 exists as a mixture of monomer, dimer, and oligomer in solution. Inter-chain salt bridges, which are not fully conserved in EcOtsA, play primary roles in MoTps1 oligomerization. Binding of UDPG by MoTps1 C-terminal domain modifies the substrate pocket of MoTps1. In the MoTps1 ternary complex structure, UDP and T6P, the products of UDPG and G6P, are detected, and substantial conformational rearrangements of N-terminal domain, including structural reshuffling (β3–β4 loop to α0 helix) and movement of a ‘shift region' towards the catalytic centre, are observed. These conformational changes render MoTps1 to a ‘closed' state compared with its ‘open' state in apo or UDPG complex structures. By solving the EcOtsA apo structure, we confirmed that similar ligand binding induced conformational changes also exist in EcOtsA, although no structural reshuffling involved. Based on our research and previous studies, we present a model for the catalytic process of Tps1. Our research provides novel information on MoTps1, Tps1 family, and structure-based antifungal drug design.

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High throughput phenotype screening pipeline for functional genomics in Magnaporthe oryzae

Protocol Exchange ◽

10.1038/nprot.2007.168 ◽

2007 ◽

Cited By ~ 1

Author(s):

Sook-Young Park ◽

Myoung-Hwan Chi ◽

Junhyun Jeon ◽

Yong-Hwan Lee

Keyword(s):

Functional Genomics ◽

High Throughput ◽

Magnaporthe Oryzae ◽

Phenotype Screening

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Antagonistic activity of Bacillus subtilis strains isolated from various sources

Ukrainian Journal of Ecology ◽

10.15421/2018_354 ◽

2018 ◽

Vol 8 (2) ◽

pp. 354-364

Author(s):

A. N. Irkitova ◽

A. V. Grebenshchikova ◽

A. V. Matsyura

Keyword(s):

Escherichia Coli ◽

Bacillus Subtilis ◽

Wild Strain ◽

Fish Farming ◽

Antagonistic Activity ◽

Antagonistic Effect ◽

E Coli ◽

Long Period ◽

Test Culture ◽

Agar Media

An important link in solving the problem of healthy food is the intensification of the livestock, poultry and fish farming, which is possible only in the adoption and rigorous implementation of the concept of rational feeding of animals. In the implementation of this concept required is the application of probiotic preparations. Currently, there is an increased interest in spore probiotics. In many ways, this can be explained by the fact that they use no vegetative forms of the bacilli and their spores. This property provides spore probiotics a number of advantages: they are not whimsical, easily could be selected, cultivated, and dried. Moreover, they are resistant to various factors and could remain viable during a long period. One of the most famous spore microorganisms, which are widely used in agriculture, is Bacillus subtilis. Among the requirements imposed to probiotic microorganisms is mandatory – antagonistic activity to pathogenic and conditional-pathogenic microflora. The article presents the results of the analysis of antagonistic activity of collection strains of B. subtilis, and strains isolated from commercial preparations. We studied the antagonistic activity on agar and liquid nutrient medias to trigger different antagonism mechanisms of B. subtilis. On agar media, we applied three diffusion methods: perpendicular bands, agar blocks, agar wells. We also applied the method of co-incubating the test culture (Escherichia coli) and the antagonist (or its supernatant) in the nutrient broth. Our results demonstrated that all our explored strains of B. subtilis have antimicrobial activity against a wild strain of E. coli, but to varying degrees. We identified strains of B. subtilis with the highest antagonistic effect that can be recommended for inclusion in microbial preparations for agriculture.

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Faculty Opinions recommendation of MoSec61β, the beta subunit of Sec61, is involved in fungal development and pathogenicity, plant immunity, and ER-phagy in Magnaporthe oryzae.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.739100266.793580524 ◽

2020 ◽

Author(s):

Karin Romisch

Keyword(s):

Magnaporthe Oryzae ◽

Plant Immunity ◽

Beta Subunit ◽

Fungal Development

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Pembuatan Media Pembelajaran Fisika Berbasis Macromedia Flash

ISLAMIKA ◽

10.36088/islamika.v1i1.184 ◽

2019 ◽

Vol 1 (1) ◽

pp. 97-114

Author(s):

Lalu A. Hery Qusyairi

Keyword(s):

Agar Media

Pembuatan media ini bertujuan untuk menghasikan media pembelajaran fisika berbasiskan komputer dengan pokok bahasan listrik dinamis menggunakan macromedia flash 8. Pada pembuatan media pembelajaran fisika ini, software yang digunakan menggunakan software utama Macromedia Flash Professional 8 dalam pembuatan gambar dan animasi. Selain software utama, diperlukan software pendukung antara lain Nero 7 Ultra Edition digunakan untuk membakar CD atau membuat software dalam kepingan CD dan paint digunakan jika software utama Macromedia Flash Professional 8 tidak dapat difungsikan. Berdasarkan tujuan dan hasil kegiatan pembuatan media maka proses pembuatannya meliputi proses prancangan dan dua tahap penyelesaian yaitu perubahan tata letak isi media dan penyempurnaan tampilan. Hal ini dilakukan agar media ini memiliki estetis yang tinggi dan kesesuain antara desain dan isi materi. Kemudian dilakukan penyimpanan atau pembakaran file ke dalam bentuk Cd outorun dengan softwere Nero 7 Ultra Edition agar media ini dapat diinstalasikan secara langsung pada komputer pada saat CD dimasukkan kedalam CDRoom. Hasil akhir sebuah softwere pembelajaran fisika dengan bantuan media komputer pada pokok bahasan listrik dinamis berbasiskan macromedia flash profesioanl 8 adalah media pembelajaran fisika dalam bentuk CD outorun yang asalah datu file utamanya berekstensi .exe.

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