scholarly journals Improved phenotyping procedure for evaluating resistance in rice against gall midge (Orseolia oryzae, Wood-Mason)

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Ling Cheng ◽  
Fugang Huang ◽  
Zhe Jiang ◽  
Baiyi Lu ◽  
Xiaohui Zhong ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Resistance Gene ◽  
High Throughput ◽  
Gall Midge ◽  
Screening Method ◽  
Molecular Techniques ◽  
Orseolia Oryzae ◽  
Mutant Screening ◽  
Biological Studies ◽  
High Throughput Phenotyping

Abstract Background The rice gall midge (RGM, Orseolia oryzae, Wood-Mason), an important stem-feeding pest worldwide, has caused serious production losses over the past decades. Rice production practices indicate that the most reliable method for managing RGM is the deployment of cultivars that incorporate host resistance. However, the conventional phenotypic screening method of rice resistance to RGM suggested by the International Rice Research Institute (IRRI) has been used for approximately 30 years, and only 12 rice varieties/lines (including controls) can be evaluated in one tray. It is not suitable for high-throughput phenotyping of rice germplasm. Moreover, a suitable method to prepare samples for molecular biological studies of rice resistance against RGM is imperative with the rapid development of modern molecular techniques. Results The proper density of seedlings/RGM was determined for four seeding arrangements. A high-throughput phenotyping method (HTPM) for 60 lines/varieties infested with 36 female RGM adults in one tray, as described by method 4–3 (seeded 60 lines/varieties), was developed and verified using mutant screening. Furthermore, one RGM resistance gene flanked by markers 12RM28346 and 12RM28739 on chromosome 12 was simultaneously detected using method 2–2 (seeded 30 lines/varieties in one tray) treated with 24 RGM and analyzed using conventional and simplified grading systems. Genetic analysis of the RGM resistance gene was confirmed using a method identical to that suggested by IRRI. Finally, one bucket with 24 seedlings treated with at least five female RGM adults was efficacious and could offer adequate samples for insect development observation or molecular biological studies. Conclusion A highly efficient and reliable procedure for evaluation of resistance in rice to RGM was developed and improved, and was verified through mutant screening, gene mapping, genetic analysis, and insect growth and development observations.

Genetic analysis and fine mapping of the gall midge resistance gene Gm5 in rice (Oryza sativa L.)

2020 ◽  
Vol 133 (6) ◽  
pp. 2021-2033 ◽  
Author(s):  
Hailian Zhou ◽  
Xinyi Wang ◽  
Yi Mo ◽  
Yang Li ◽  
Liuhui Yan ◽  
...  
Keyword(s):  
Oryza Sativa ◽  
Genetic Analysis ◽  
Resistance Gene ◽  
Fine Mapping ◽  
Oryza Sativa L ◽  
Gall Midge

scholarly journals Identification of a new resistance locus against the rice gall midge, Orseolia oryzae (Wood-Mason), in Thai rice landrace MN62M

2019 ◽  
Author(s):  
Phikul Leelagud ◽  
Sakda Kongsila ◽  
Phanchita Vejchasarn ◽  
Kulchana Darwell ◽  
Yotwarit Phansenee ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Ssr Markers ◽  
Resistance Genes ◽  
Gall Midge ◽  
Rice Chromosome ◽  
Chromosome 2 ◽  
Orseolia Oryzae ◽  
Rice Varieties ◽  
Rice Gall Midge ◽  
Rice Landrace

Abstract Background The rice gall midge (RGM), Orseolia oryzae (Wood-Mason), is one of the most destructive insect pests of rice, and it causes significant yield losses annually in Asian countries. The development of resistant rice varieties is considered as the most effective and economical approach for maintaining yield stability by controlling RGM. Identification of resistance genes will help in marker-assisted selection (MAS) to pyramid the resistance genes and develop a durable resistance variety against RGM in areas with frequent outbreaks.Results A mitochondrial gene, cytochrome C oxidase I (COI), was used to analyze the genetic diversity among Thai RGM populations. The phylogenetic tree indicated that the Thai RGM populations were homogeneously distributed throughout the country, except for some populations in central and northeast Thailand that probably became isolated from the main population. The reactions of the resistant rice varieties carrying different resistance genes revealed different RGM biotypes in Thailand. The Thai rice landrace MN62M showed resistance to all RGM populations used in this study. We identified a novel genetic locus for resistance to RGM, designated as GM12 , on the short arm of rice chromosome 2. The locus was identified using linkage analysis in 144 F 2 plants derived from a cross between susceptible cultivar KDML105 and RGM-resistant cultivar MN62M with single nucleotide polymorphism (SNP) markers and F 2:3 phenotype. The locus was confirmed and mapped using SNP and simple sequence repeat (SSR) markers surrounding the target chromosomal location. Finally, the locus was mapped between two flanking markers, RM6800 and S2_419160.Conclusions We identified a new RGM resistance gene, GM12 , on rice chromosome 2 in the Thai rice landrace MN62M. This finding yielded SNP and SSR markers that can be used in MAS to develop cultivars with broad-spectrum resistance to RGM. The new resistance gene provides important information for the identification of RGM biotypes in Thailand and Southeast Asia.

Towards high-throughput phenotyping of complex patterned behaviors: Lessons from mouse self-grooming

2011 ◽  
Author(s):  
E. Kyzar ◽  
S. Gaikwad ◽  
M. Pham ◽  
J. Green ◽  
A. Roth ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Phenotyping

Directed Evolution of P450 Fatty Acid Decarboxylases via High-Throughput Screening Towards Improved Catalytic Activity

2019 ◽  
Author(s):  
Huifang Xu ◽  
Weinan Liang ◽  
Linlin Ning ◽  
Yuanyuan Jiang ◽  
Wenxia Yang ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Fatty Acid ◽  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening ◽  
Colorimetric Detection ◽  
Screening Method ◽  
Random Mutagenesis ◽  
Direct Production ◽  
Consumption Activity

P450 fatty acid decarboxylases (FADCs) have recently been attracting considerable attention owing to their one-step direct production of industrially important 1-alkenes from biologically abundant feedstock free fatty acids under mild conditions. However, attempts to improve the catalytic activity of FADCs have met with little success. Protein engineering has been limited to selected residues and small mutant libraries due to lack of an effective high-throughput screening (HTS) method. Here, we devise a catalase-deficient <i>Escherichia coli</i> host strain and report an HTS approach based on colorimetric detection of H<sub>2</sub>O<sub>2</sub>-consumption activity of FADCs. Directed evolution enabled by this method has led to effective identification for the first time of improved FADC variants for medium-chain 1-alkene production from both DNA shuffling and random mutagenesis libraries. Advantageously, this screening method can be extended to other enzymes that stoichiometrically utilize H<sub>2</sub>O<sub>2</sub> as co-substrate.

scholarly journals Development of a Novel High-Throughput Screen and Identification of Small-Molecule Inhibitors of the Gα–RGS17 Protein–Protein Interaction Using AlphaScreen

2011 ◽  
Vol 16 (8) ◽  
pp. 869-877 ◽  
Author(s):  
Duncan I. Mackie ◽  
David L. Roman
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Protein Interaction ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Screening Method ◽  
Fold Increase ◽  
Rgs Proteins ◽  
High Throughput Screen ◽  
Protein Protein Interaction

In this study, the authors used AlphaScreen technology to develop a high-throughput screening method for interrogating small-molecule libraries for inhibitors of the Gαo–RGS17 interaction. RGS17 is implicated in the growth, proliferation, metastasis, and the migration of prostate and lung cancers. RGS17 is upregulated in lung and prostate tumors up to a 13-fold increase over patient-matched normal tissues. Studies show RGS17 knockdown inhibits colony formation and decreases tumorigenesis in nude mice. The screen in this study uses a measurement of the Gαo–RGS17 protein–protein interaction, with an excellent Z score exceeding 0.73, a signal-to-noise ratio >70, and a screening time of 1100 compounds per hour. The authors screened the NCI Diversity Set II and determined 35 initial hits, of which 16 were confirmed after screening against controls. The 16 compounds exhibited IC50 <10 µM in dose–response experiments. Four exhibited IC50 values <6 µM while inhibiting the Gαo–RGS17 interaction >50% when compared to a biotinylated glutathione-S-transferase control. This report describes the first high-throughput screen for RGS17 inhibitors, as well as a novel paradigm adaptable to many other RGS proteins, which are emerging as attractive drug targets for modulating G-protein-coupled receptor signaling.

scholarly journals A high-throughput screening method for evolving a demethylase enzyme with improved and new functionalities

2020 ◽  
Author(s):  
Yuru Wang ◽  
Christopher D Katanski ◽  
Christopher Watkins ◽  
Jessica N Pan ◽  
Qing Dai ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Targeted Mutagenesis ◽  
Rna Repair ◽  
Dna And Rna ◽  
Modified Dna ◽  
Dna Substrates ◽  
Trna Sequencing

Abstract AlkB is a DNA/RNA repair enzyme that removes base alkylations such as N1-methyladenosine (m1A) or N3-methylcytosine (m3C) from DNA and RNA. The AlkB enzyme has been used as a critical tool to facilitate tRNA sequencing and identification of mRNA modifications. As a tool, AlkB mutants with better reactivity and new functionalities are highly desired; however, previous identification of such AlkB mutants was based on the classical approach of targeted mutagenesis. Here, we introduce a high-throughput screening method to evaluate libraries of AlkB variants for demethylation activity on RNA and DNA substrates. This method is based on a fluorogenic RNA aptamer with an internal modified RNA/DNA residue which can block reverse transcription or introduce mutations leading to loss of fluorescence inherent in the cDNA product. Demethylation by an AlkB variant eliminates the blockage or mutation thereby restores the fluorescence signals. We applied our screening method to sites D135 and R210 in the Escherichia coli AlkB protein and identified a variant with improved activity beyond a previously known hyperactive mutant toward N1-methylguanosine (m1G) in RNA. We also applied our method to O6-methylguanosine (O6mG) modified DNA substrates and identified candidate AlkB variants with demethylating activity. Our study provides a high-throughput screening method for in vitro evolution of any demethylase enzyme.

scholarly journals High-throughput phenotyping: breaking through the bottleneck in future crop breeding

2021 ◽  
Author(s):  
Peng Song ◽  
Jinglu Wang ◽  
Xinyu Guo ◽  
Wanneng Yang ◽  
Chunjiang Zhao
Keyword(s):  
High Throughput ◽  
Crop Breeding ◽  
High Throughput Phenotyping
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