2018 Fiscal Year Annual Research Report
Functional characterization of candidate genes for PUE and mining their allelic variants in rice
| Project/Area Number |
18F18703
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| Research Institution | Japan International Research Center for Agricultural Sciences |
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Principal Investigator |
Wissuwa Matthias 国立研究開発法人国際農林水産業研究センター, 生産環境・畜産領域, 主任研究員 (90442722)
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| Co-Investigator(Kenkyū-buntansha) |
ADEM GETNET 国立研究開発法人国際農林水産業研究センター, 生産環境・畜産領域, 外国人特別研究員
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| Project Period (FY) |
2018-07-25 – 2021-03-31
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| Keywords | expression / bioinformatics / QPCR / Gene 13 / Gene 14 / Gene 30 / PM-H+ATP / remobilization P |
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| Outline of Annual Research Achievements |
In 2018 budget year, one experiment was completed. This experiment studied the expression level of genes which were found by bioinformatics gene prediction tool from novel loci identified for Phosphorus utilization efficiency in our lab with the aim of identifying candidate genes from these loci. Seven predicted genes were selected from the 80 predicted genes. To see if these predicted genes are found in our experimental genotypes, PCR in both root and leaf tissues were done. Positive PCR bands were found for Gene 13, Gene 14, Gene 30 and PM-H+ATPase. These genes were sequenced and the sequence were aligned with bioinformatics tool and QPCR primer was designed and QPCR analyses were made. Second large experiment which is split into two major parts is underway together with another JSPS fellow (Hayes Patrick). One part studies about the remobilization P in the course of vegetative growth including the gene expression of our candidate genes/known remobilization genes and the redistribution of P into different metabolites and inorganic P including the gene expression of candidate genes/known P redistribution genes in relation to this research question. Most part of the physiological data and part of the genetic data is already collected and it will be ongoing for 2019 budget year.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Reasons- One experiment completed, another on going and new experiment is planned.
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| Strategy for Future Research Activity |
The second large experiment, which has two parts, is ongoing in 2019 budget year. For the remobilization experiment, tissue digestion and subsequent P analysis will be performed. RNA extraction (> 400 samples) and cDNA synthesis will be done. A manageable number of genes will be selected based on our physiological data from both parts of these experiments and QPCR analysis will be conducted on these selected genes. Once all the necessary data has been collected, data analysis will be performed and Journal article will be drafted, finalized and submitted to peer-reviewed reputable Journal.
From the completed experiment in 2018 budget year, a new experiment may arise for 2019 and 2020 budget year. This experiment may focus on mutants of the genes identified by gene prediction from the novel loci identified in our lab previously and which were further tested for their expression in our experimental genotypes and showed reduced expression in most tissues and genotypes. This has changed the nature of the project plan submitted for JSPS to some extent, focusing on T-DNA insertion mutants rather than over- and under- expression of candidate genes. Finally, in the middle or around the completion of my fellowship, I am planning to attend conference.
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