Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/35849
標題: 菌質體之分泌蛋白在擬南芥中調節缺磷反應與防禦機制的探討
The Phytoplasma Effector Modulates Phosphate Homeostasis and Defense Response in Arabidopsis
作者: 呂晏婷
Lu, Yen-Ting
關鍵字: 菌質體
phytoplasma
翠菊黃萎病
作用因子
缺磷反應
免疫反應
effector
Pi deficiency
defense response
出版社: 生物科技學研究所
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摘要: Phytoplasmas are wall-less bacterial plant pathogens. They are restricted to the cytoplasm of phloem sieve cells, and can not be cultured in artificial culture medium. Phytoplasmas are spread by phloem-feeding insects including leafhoppers, planthoppers and psyllids. The disease symptoms caused by phytoplasmas including yellowing leaf, greening flowers, proliferation of stems and phyllody. Those morphological changes are mainly caused by the secreted effectors produced by phytoplasmas. The whole genome of Aster Yellows phytoplasma strain witches’ broom (AY-WB) has been completely sequenced, and 56 candidate effectors has been identified and named secreted AY-WB proteins (SAPs). Among them, SAP11 contains a nuclear localization signal (NLS) for nucleus targeting. SAP11 has been shown to destabilize plant TCP transcription factor, in order to repress the expression of LIPOXYGENASE2, an essential enzyme for jasmonate (JA) biosynthesis. Although SAP11 is a small protein with a molecular weight of approximately 14 kDa, the molecular mechanism in disease developing is still not clear. Here, we found that the purified recombinant SAP11 protein was unstable and could not be crystallized. However, after gel-filtration chromatography analysis, we showed that SAP11 formed a multimeric protein. With RNA-seq analysis, we showed that the expression of Pi deficiency-induced genes such as IPS1, PS2, PHT1;4, IPS2... were triggered by SAP11. Because Pi is an essential element for intracellular pathogens, the regulation of Pi homeostasis may contribute to the growth of phytoplasmas in host cells. In addition, we found that the expression of defense response genes such as PR1, WIN3, PAD4… were also suppressed by SAP11. This data suggests that phytoplasmas repress plant immune responses via secreted SAP11.
植物菌質體(phytoplasma)為一種無細胞壁植物病原菌,寄生在宿主篩管細胞內,且至今尚無法以人工方式培養。菌質體主要藉由葉蟬、飛蝨等媒介昆蟲吸食植物韌皮部組織汁液而傳播。菌質體感染的植株具有發育遲緩、植株矮化、節間縮短、莖葉叢生、葉片捲曲、黃化、花器綠化或葉化等外表型態的改變。目前已知這些病徵的表現和菌質體分泌的作用因子(effector)有關。本實驗研究的材料為翠菊黃萎病(aster yellows)中發現的植物菌質體(Aster Yellows phytoplasma strain Withches’ Broom,AY-WB)。此菌質體的序列已被解序,而其基因組可轉譯出56個分泌性的AY-WB蛋白,簡稱SAPs。SAP11為其中之一,帶有細胞核定位信號(nuclear localization signals,NLSs)且可進入宿主細胞的細胞核,會促進TCP轉錄因子(TCP transcription factors)的降解,進而抑制LOX2的表現,使茉莉酸(jasmonate,JA)生合成量下降。SAP11大小約14 kDa,致病機制尚未清楚。因此本實驗期望能藉不同的實驗方法以更進一步了解SAP11的功能。首先在大腸桿菌中表現重組SAP11蛋白並純化後,藉由分子篩選層析管柱分析可知SAP11會形成聚分子。但因蛋白分子結構不穩定,不易形成結晶。進一步利用次世代定序方式進行SAP11轉殖株與野生種擬南芥的轉錄體分析,發現SAP11的表現會誘發植物產生缺磷反應,其中IPS1、PS2、PHT1;4、IPS2、…等缺磷誘導的基因皆大量表現。由於磷是許多病原菌的必需生存元素,因此調控磷的吸收可能有助於菌質體的繁殖。另一方面,發現表達SAP11會抑制植物產生免疫反應,PR1、WIN3、PAD4、…等水楊酸相關基因有表現量降低或表現時間延後的現象。而這與先前研究指出表現SAP11可提高菌質體昆蟲宿主繁殖後代量之結果相符。因此推測菌質體可能透過分泌SAP11到植物體內降低免疫反應,以提高自身以及傳播之昆蟲宿主繁殖與存活率。
URI: http://hdl.handle.net/11455/35849
其他識別: U0005-2208201316261000
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2208201316261000
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