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Title: | 番茄初級免疫相關基因座Bwr12內兩個LRR蛋白質之研究 Study on two LRR proteins in the tomato Bwr12 locus involved in innate immunity |
Authors: | Chun-You Kuo 郭俊佑 |
Advisor: | 鄭秋萍(Chiu-Ping Cheng) |
Keyword: | 番茄,青枯病菌,PTI,Bwr12,LRR,ACO,CYP, tomato,Ralstonia solanacearum,PTI,Bwr12,LRR,ACO,CYP, |
Publication Year : | 2021 |
Degree: | 碩士 |
Abstract: | 由Ralstonia solanacearum (Rs) 引起的青枯病 (bacterial wilt, BW) 是許多重要經濟作物的嚴重病害,抗病育種是防治番茄BW的重要手段。目前具穩定抗BW的番茄品種Hawaii 7996(H7996)之多個數量性狀位點 (QTLs) 雖已定位,然而其涉及之分子機制與主導之防禦基因仍未知。本研究室先前未發表之研究指出H7996具有強烈的廣效初級免疫PAMP-triggered immunity (PTI),且抗第一型Rs (phylotype I) 之主控QTL Bwr12也參與其PTI反應。本研究旨在深入探討在不同番茄品系中,Bwr12基因座內兩個LRR蛋白質12g550與12g520所參與之PTI調控分子機制。在BW抗病與感病品種中12g550與12g520之編碼序列 (coding sequence) 並無差異,但在抗病品種H7996中12g550基因啟動子序列除了有些許 single nucleotide polymorphism (SNPs) 外也短少了308 bp,而12g520基因啟動子序列有兩個SNPs,雖然目前的基因啟動子活性分析初步結果尚無一致結果,或許這些啟動子序列差異造成與不同轉錄因子(transcription factors) 結合進而可能調控基因差異表現。此外,不論有或無Rs感染,在抗病與感病品種中這兩個基因被預測的intron皆存在其轉錄產物中,且導致12g550轉譯提早終止而產生僅有93個胺基酸之蛋白質產物。NbACO2a與NbCYP6為利用共免疫沈澱與奈流液相層析串連二次質譜 (Co-IP-LC-MS/MS) 篩選出12g5501-93AA之可能互作蛋白質,而目前基因功能分析結果顯示12g520、12g5501-93AA、NbACO2a及NbCYP6在菸草PTI反應中均為正向調控者。預期本研究將有助於提供番茄抗青枯病與PTI之關鍵訊息。 Bacterial wilt (BW) caused by Ralstonia solanacearum (Rs) is a global destructive disease of many crops. Tomato cultivar Hawaii 7996 (H7996) is a reliable resistance source against BW; however, the mechanisms of H7996 defense remained undetermined. Our previous studies showed that H7996 displays strong pattern-triggered immunity (PTI) responses, and that the major QTL associated with its BW-resistance, Bwr12, is involved in PTI. This study aimed to comparatively investigate the mechanisms underlying tomato PTI mediated by two LRR genes located in Bwr12, 12g520 and 12g550, in different tomato cultivars. The coding sequences of 12g520 and 12g550 in BW-resistant and -susceptible cultivars are identical. However, in H7996, the 12g550 promoter has a 308-bp deletion and 12g520 promoter contains two single-nucleotide-polymorphisms (SNPs). Although preliminary results of promoter activity assays was not consistent with the transcriptional expressions of these two genes, the differences of predicted transcription factors of these sequences might relate to the differential expression patterns. Furthermore, the predicted introns are present in the transcripts of these genes in both BW-resistant and -susceptible cultivars without or with Rs infection, possibly leading to production of a 12g550 protein product of 93 amino acids due to an early termination in translation. NbACO2a and NbCYP6 are candidate interacting proteins of 12g5501-93AA identified by Co-IP-LC-MS/MS in Nicotiana benthamiana. Current data of functional genetic studies revealed positive roles of 12g520, 12g5501-93AA, NbACO2a and NbCYP6 play positive roles in PTI responses. These results are expected to shed light on plant BW-defense and PTI. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/20842 |
DOI: | 10.6342/NTU202100709 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 植物科學研究所 |
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U0001-1602202122511400.pdf Restricted Access | 9.63 MB | Adobe PDF |
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