scholarly journals An Efficient Genetic Manipulation Protocol for Dark Septate Endophyte Falciphora Oryzae

2021 ◽  
Author(s):  
Zhen-Zhu Su ◽  
Meng-Di Dai ◽  
Jia-Nan Zhu ◽  
Yu-Lan Zeng ◽  
Xuan-Jun Lu ◽  
...  
Keyword(s):  
Endophytic Fungus ◽  
Plant Biomass ◽  
Genetic Manipulation ◽  
Blast Resistance ◽  
Enzyme Digestion ◽  
Dark Septate Endophyte ◽  
Rice Blast Resistance ◽  
Rice Roots ◽  
Manipulation System ◽  
Protoplast Preparation

Abstract Falciphora oryzae is a dark septate endophyte (DSE) isolated from wild rice roots (Oryza sativa L.). It was classified as a non-clavicitaceous endophyte. The fungus colonizes rice roots, showing a significant increase in agronomic parameters with plant biomass, rice blast resistance, yield, and quality. The construction of the genetic manipulation system is critical to study the relationship between F. oryzae and O. sativa. In the present study, the protoplast preparation and transformation system of F. oryzae was investigated. The key parameters affecting the efficiency of protoplast production, such as osmotic pressure stabilizer, enzyme digestion conditions, and fungal age, were studied. The results showed that F. oryzae strain obtained higher protoplast yield and effective transformation when treated with enzyme digestion solution containing 0.9mol L-1 KCl solution and 10 mg mL−1 glucanase at 30℃ with shaking 80 rpm for 2-3 h. When the protoplasts were plated on a regenerations-agar (RgA) medium containing 1M sucrose, the re-growth rate of protoplasts was the highest. We successfully acquired GFP-expressing transformants by transforming the pKD6-GFP vector into protoplasts. Further, the GFP expression in fungal hyphae possessed good stability and intensity during symbiosis in rice roots.The genetic manipulation system of endophytic fungus facilitates the further exploration the interaction between the endophytic fungus and their hosts.

scholarly journals Natural Variation at the Pi-ta Rice Blast Resistance Locus

2003 ◽  
Vol 93 (11) ◽  
pp. 1452-1459 ◽  
Author(s):  
Yulin Jia ◽  
Gregory T. Bryan ◽  
Leonard Farrall ◽  
Barbara Valent
Keyword(s):  
Positional Cloning ◽  
Magnaporthe Grisea ◽  
Blast Resistance ◽  
El Paso ◽  
Avirulence Gene ◽  
Resistance Locus ◽  
Japonica Rice ◽  
Breeding Programs ◽  
Rice Blast Resistance ◽  
Intron Region

The resistance gene Pi-ta protects rice crops against the fungal pathogen Magnaporthe grisea expressing the avirulence gene AVR-Pita in a gene-for-gene manner. Pi-ta, originally introgressed into japonica rice from indica origin, was previously isolated by positional cloning. In this study, we report the nucleotide sequence of a 5,113-base pair region containing a japonica susceptibility pi-ta allele, which has overall 99.6% nucleotide identity to the indica Pi-ta allele conferring resistance. The intron region shows the levels of sequence diversity that typically differentiate genes from indica and japonica rices, but the other gene regions show less diversity. Sequences of the Pi-ta allele from resistant cultivars Katy and Drew from the southern United States are identical to the resistance Pi-ta sequence. Sequences from susceptible cultivars El Paso 144 and Cica 9 from Latin America define a third susceptibility haplotype. This brings the total number of Pi-ta haplotypes identified to four, including the resistance allele and three susceptibility alleles. The Pi-ta locus shows low levels of DNA polymorphism compared with other analyzed R genes. Understanding the natural diversity at the Pi-ta locus is important for designing specific markers for incorporation of this R gene into rice-breeding programs.

scholarly journals Rice blast resistance-inducing mechanisms via effector recognition by NLR immune receptors

2016 ◽  
Vol 82 (4) ◽  
pp. 296-300 ◽  
Author(s):  
H. SAITOH ◽  
H. KANZAKI ◽  
K. FUJISAKI ◽  
H. TAKAGI ◽  
K. YOSHIDA ◽  
...  
Keyword(s):  
Rice Blast ◽  
Blast Resistance ◽  
Rice Blast Resistance ◽  
Immune Receptors ◽  
Effector Recognition

scholarly journals Development of an efficient conjugation-based genetic manipulation system for Pseudoalteromonas

2015 ◽  
Vol 14 (1) ◽  
pp. 11 ◽  
Author(s):  
Pengxia Wang ◽  
Zichao Yu ◽  
Baiyuan Li ◽  
Xingsheng Cai ◽  
Zhenshun Zeng ◽  
...  
Keyword(s):  
Genetic Manipulation ◽  
Manipulation System

Marker Development for Rice Blast Resistance Gene Pi66(t) and Application in the USDA Rice Mini-Core Collection

Crop Science ◽  
2016 ◽  
Vol 56 (3) ◽  
pp. 1001-1008 ◽  
Author(s):  
Yan Liu ◽  
Yulin Jia ◽  
David Gealy ◽  
David M. Goad ◽  
Ana L. Caicedo ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Rice Blast ◽  
Core Collection ◽  
Blast Resistance ◽  
Marker Development ◽  
Rice Blast Resistance ◽  
Blast Resistance Gene ◽  
Mini Core Collection

Expression-based genotyping of the rice blast resistance genes in the elite maintainer line Yixiang1B

2017 ◽  
Vol 148 (4) ◽  
pp. 955-965 ◽  
Author(s):  
Li Wang ◽  
Xiao-Hong Hu ◽  
Gang Lin ◽  
De-Ming Zhao ◽  
Jun Shi ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Rice Blast ◽  
Blast Resistance ◽  
Maintainer Line ◽  
Rice Blast Resistance ◽  
Blast Resistance Genes

scholarly journals Osa-miR162a balances rice yield and resistance to Magnaporthe oryzae

2020 ◽  
Author(s):  
Xu-Pu Li ◽  
Xiao-Chun Ma ◽  
He Wang ◽  
Yong Zhu ◽  
Xin-Xian Liu ◽  
...  
Keyword(s):  
Magnaporthe Oryzae ◽  
Rice Blast ◽  
Rice Yield ◽  
Blast Resistance ◽  
Blast Disease ◽  
Mirna Biogenesis ◽  
Trade Off ◽  
Rice Blast Resistance ◽  
Defense Related Gene ◽  
Transgenic Lines

Abstract MicroRNAs (miRNAs) play essential roles in rice immunity against Magnaporthe oryzae, the causative agent of rice blast disease. Osa-miR162a targets Dicer-like 1 (DCL1) genes, which play vital roles in miRNA biogenesis and act as negative regulators in rice immunity. Here we demonstrate that Osa-miR162a improves rice immunity against M. oryzae and balances the trade-off between rice yield and resistance. Overexpression of Osa-miR162a compromises rice susceptibility to M. oryzae accompanying enhanced induction of defense-related genes and accumulation of hydrogen peroxide (H2O2). In contrast, blocking miR162 by overexpressing a target mimic of miR162 enhances susceptibility to blast fungus associating with compromised induction of defense-related gene expression and H2O2 accumulation. Moreover, the transgenic lines overexpressing Osa-miR162a display decreased seed setting rate resulting in reduced yield per plant, whereas blocking miR162 leads to an increased number of grains per panicle, resulting in increased yield per plant. Altered accumulation of miR162 had limited impact on the expression of OsDCL1. Together, our results indicate that Osa-miR162a improves rice blast resistance and plays a role in the balance of trade-off between resistance and yield.

scholarly journals Screening of the Dominant Rice Blast Resistance Genes with PCR-based SNP and CAPS Marker in Aromatic Rice Germplasm

2011 ◽  
Vol 56 (4) ◽  
pp. 329-341 ◽  
Author(s):  
Jeong-Soon Kim ◽  
Sang-Nag Ahn ◽  
Sung-Jun Hong ◽  
Jin-Hyeuk Kwon ◽  
Yeong-Ki Kim ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Rice Blast ◽  
Blast Resistance ◽  
Aromatic Rice ◽  
Caps Marker ◽  
Rice Blast Resistance ◽  
Rice Germplasm ◽  
Blast Resistance Genes

scholarly journals Application of CRISPR/Cas9 Tools for Genome Editing in the White-Rot Fungus Dichomitus squalens

Biomolecules ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1526
Author(s):  
Joanna E. Kowalczyk ◽  
Shreya Saha ◽  
Miia R. Mäkelä
Keyword(s):  
Genome Editing ◽  
Plant Biomass ◽  
Genetic Manipulation ◽  
White Rot ◽  
White Rot Fungus ◽  
Detailed Knowledge ◽  
Wild Type ◽  
Low Frequencies ◽  
Dichomitus Squalens

Dichomitus squalens is an emerging reference species that can be used to investigate white-rot fungal plant biomass degradation, as it has flexible physiology to utilize different types of biomass as sources of carbon and energy. Recent comparative (post-) genomic studies on D. squalens resulted in an increasingly detailed knowledge of the genes and enzymes involved in the lignocellulose breakdown in this fungus and showed a complex transcriptional response in the presence of lignocellulose-derived compounds. To fully utilize this increasing amount of data, efficient and reliable genetic manipulation tools are needed, e.g., to characterize the function of certain proteins in vivo and facilitate the construction of strains with enhanced lignocellulolytic capabilities. However, precise genome alterations are often very difficult in wild-type basidiomycetes partially due to extremely low frequencies of homology directed recombination (HDR) and limited availability of selectable markers. To overcome these obstacles, we assessed various Cas9-single guide RNA (sgRNA) ribonucleoprotein (RNP) -based strategies for selectable homology and non-homologous end joining (NHEJ) -based gene editing in D. squalens. We also showed an induction of HDR-based genetic modifications by using single-stranded oligodeoxynucleotides (ssODNs) in a basidiomycete fungus for the first time. This paper provides directions for the application of targeted CRISPR/Cas9-based genome editing in D. squalens and other wild-type (basidiomycete) fungi.

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