鑑定及分析與缺乏營養相關之水稻突變株;Identification and Characterization of Rice Mutants Related to Nutrient Starvation in Rice (Oryza sativa)

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題名:	鑑定及分析與缺乏營養相關之水稻突變株;Identification and Characterization of Rice Mutants Related to Nutrient Starvation in Rice (Oryza sativa)
作者:	黎氏裝;Trang, Le Thi
貢獻者:	生命科學系
關鍵詞:	水稻;Rice;Transgenic;Rice Genomic Function;Nutrients response
日期:	2019-07-31
上傳時間:	2019-09-03 14:33:29 (UTC+8)
出版者:	國立中央大學
摘要:	水稻是一種重要的作物，為世界近50％的人口提供食物。雖然已經開發出先進技術來提高水稻產量，但增加水稻產量仍然是研究人員的主要目標。水稻的營養利用效率對高產是必不可少的。為了確定涉及營養利用過程的因素，在氮（N），磷酸鹽（P）和碳（C）飢餓條件下生長的水稻中進行微陣列分析。選擇在N，P，C-飢餓條件下具有改變的mRNA積累的總共32個基因用於進一步研究。從台灣水稻插入突變體（TRIM）群體中收集總共69個對應於這些基因的T-DNA插入突變體用於進一步研究。篩選後，在稻田中培養38個T2代突變體系，用於檢查T3種子的表型和繁殖。許多突變體系顯示表型，例如晚熟，矮化，黃葉，程序性細胞死亡，直立葉減少生長，更高的穀物產量和更低的穀物產量。已經進行了基因分型以確定突變表型是否與靶基因上的T-DNA插入相關。在溫室和稻田中培養直立葉和黃葉突變體以確認表型和基因型的相關性。使用RT-PCR分析確定顯示黃色葉和直立葉的突變體周圍基因的表達。分離兩個基因LOC_Os03g53630和LOC_Os03g53650，其在黃葉突變體中具有增加的表達，以產生過表達株系。發現LOC_Os03g53640的表達在純合植物中降低，這與隱性黃葉突變體表型相關。 LOC_Os03g53640是在擬南芥中發現的藍細菌和低PSII積累中發現的光系統II 11kDa蛋白的同源物。 LOC_Os03g53640的敲除將在未來使用CRISPR / Cas 9技術進行。;Rice is an important crop that provides food for nearly 50% of the world’s population. Although advanced technologies have been developed to enhance rice yield, increasing rice yield still is the main goal for the researcher. Nutrition use efficiency of rice is essential for high yield. To identify factors involving in the nutrition utilization process, microarray analyses were performed in rice grown under nitrogen (N), phosphate (P) and carbon (C) starvation conditions. A total of 32 genes with altered mRNA accumulation under N, P, C- starvation conditions were selected for further study. A total of 69 T-DNA insertion mutants corresponding to those genes were collected from the Taiwan Rice Insertional Mutant (TRIM) population for further investigation. After screening, 38 T2 generation mutant lines were grown in the rice paddy for the examination of phenotypes and propagation of T3 seeds. Numerous mutant lines showed phenotypes such as late maturation, dwarf, yellow leaf, programmed cell death, erect leaf reduced-growth, higher grain yield, and lower grain yield. Genotyping has been carried out to determine whether mutant phenotype correlates with the T-DNA insertion on the target genes. Erect leaf and yellow leaf mutants were grown in the greenhouse and in the rice paddy to confirm the correlation of phenotype and genotype. Expression of genes surrounding mutant showing yellow leaf and erect leaf were determined using the RT-PCR analysis. Two genes, LOC_Os03g53630 and LOC_Os03g53650 with increased expression in the yellow leaf mutant, were isolated to generate overexpression lines. The expression of LOC_Os03g53640 was found to be decreased in the homozygous plant, which is correlated with the recessive yellow leaf mutant phenotype. LOC_Os03g53640 is a homolog of Photosystem II 11kDa protein found in Cyanobacteria and Low PSII accumulation19 found in Arabidopsis. Knock-out of LOC_Os03g53640 will be performed using the CRISPR/Cas 9 technology in the future.
顯示於類別:	[生命科學研究所 ] 博碩士論文

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