高溫下番茄花粉數量及花粉活性之數量性狀基因座定位以及轉錄體分析

QTL Mapping of Total Pollen Number and Pollen Viability and Expression Profiling Analysis under Moderate Heat Stress in Tomato

方昱富(Yu-Fu Fang)

指導教授 ： 陳凱儀

國立臺灣大學/生物資源暨農學院/農藝學研究所/碩士(2018年)

https://doi.org/10.6342/NTU201802585

摘要

番茄生長於高溫環境下，結果率會大幅度降低造成產量大幅下降。其中，花粉數量以及花粉活性為影響結果率之重要因子。本篇研究中使用親本為不耐熱品系 CA4 以及耐熱品系 CLN1621N 所建立之重組自交族群，共包含78個重組自交系，調查此族群於高溫逆境下 (日夜溫 30/25 度) 之花粉數量以及花粉活性。外表型資料顯示，此兩性狀與日照強度有明顯負相關，且兩性狀的廣義遺傳率介於0.3到0.6之間。為了瞭解於高溫下調控此兩性狀之遺傳因子，我們使用次世代核酸定序技術，以數量性狀基因座定位與基因表現量剖析 (即轉錄體) 兩種策略進行研究。單核苷酸分子標記的發掘來自重組自交族群的轉錄體定序資料。我們共獲得 7,550 個單核苷酸分子標記，用以建立高密度連鎖圖譜，其長度為 693.8 cM。數量性狀基因座定位分析共發現五個於高溫下調控花粉數量或活性之數量性狀基因座，分別位於第3、4、10以及11號染色體上，各別基因座可解釋的外表型變異介於 8.74% 到 34.63% 之間。此外，位於第4號染色體上影響花粉總數變異的數量性狀基因座內，14個基因之表現量高低與此染色體區間內分子標記之基因型具有統計意義上的顯著相關性。這些基因為高溫下維持花粉數量之候選基因。

關鍵字

番茄 ； 高溫 ； 花粉 ； 數量性狀基因座 ； 轉錄體分析

並列摘要

High temperature reduces the fruit set rate of tomato, so does the yield. The total pollen number and the pollen viability were recognized as two major factors involving in the reduction of the fruit set rate. In this study, we measured these two traits of heat-sensitive variety CA4, heat-tolerant variety CLN1621N and their offspring population, 78 F7 recombinant inbred lines (RIL), under 30/25 °C day/night temperature. The results showed these two traits were highly correlated to the light intensity and their broad-sense heritability were from 0.3 to 0.6. In order to understand the genetic architectures of total pollen number and pollen viability under heat stress, we deployed two strategies, genetic mapping and gene expression profiling (ie. transcriptome), using next-generation sequencing technologies. The RNAseq gene profiling data of the RIL population were used to discover single nucleotide polymorphism (SNP) markers. A total of 7,550 SNP markers were identified in this RIL population to construct the high-density genetic map which was consisted of 330 SNP markers with 693.8 cM length. The genetic mapping analyses revealed five quantitative trait loci (QTLs) on chromosome 3, 4, 10 and 11. Each QTL can explain 8.74 % to 34.63% of total variance. In addition, the variation of gene expression of 14 genes in the QTL of the total pollen number on chromosome 4 showed statistically significant association with distinct marker genotypes of this QTL. These 14 genes are the candidate genes to maintain total pollen number under heat stress.

並列關鍵字

Tomato ； Heat stress ； Pollen ； Quantitative trait loci ； Transcriptome

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