光敏色素互作因子（PIFs）廣泛存在于苔蘚到被子植物中隆敢，并在植物從暗形態(tài)建成到光形態(tài)建成的轉(zhuǎn)化過程中發(fā)揮關(guān)鍵調(diào)控作用。PIFs直接與靶基因啟動子區(qū)域的順式作用元件結(jié)合穴墅，調(diào)控基因表達(dá)，或者與其它調(diào)控因子形成復(fù)合物皇钞，共同調(diào)控靶基因表達(dá)誉结，PIFs是基因轉(zhuǎn)錄調(diào)控網(wǎng)絡(luò)中的核心，通過整合內(nèi)部和外部的信號以實(shí)現(xiàn)對植物體生長發(fā)育的調(diào)控掉盅。植物激素脫落酸（Abscisic Acid以舒，ABA）調(diào)控植物的生長發(fā)育和對逆境脅迫的響應(yīng)。ABA信號通路的很多組分已經(jīng)被鑒定出來永票。

越來越多的證據(jù)證明滥沫，ABA與PIFs轉(zhuǎn)錄因子家族在光信號通路兰绣、植物生長發(fā)育和環(huán)境適應(yīng)性方面具有協(xié)同作用。但是PIFs轉(zhuǎn)錄因子調(diào)控ABA信號轉(zhuǎn)導(dǎo)的具體機(jī)制并不清楚缀辩。

2020年4月臀玄，Biochemical and Biophysical Research Communications發(fā)表了題為“ Phytochrome-interacting factors regulate seedling growth through ABA signaling”的文章，為揭示PIFs轉(zhuǎn)錄因子調(diào)控ABA信號轉(zhuǎn)導(dǎo)機(jī)制提供了重要線索荣恐。

研究結(jié)果

1. PIFs轉(zhuǎn)錄因子在擬南芥各器官中廣泛表達(dá)并且受到ABA調(diào)控

通過RT-qPCR發(fā)現(xiàn)PIF1, PIF3, PIF4和PIF5在多種組織器官和發(fā)育階段均有表達(dá)叠穆。ABA處理后畦浓，PIF1, PIF3和PIF4的表達(dá)水平升高，但是PIF5的表達(dá)受到抑制祷嘶。

2.?pif突變體在幼苗生長期對脫落酸不敏感

首先構(gòu)建了pif1, pif3, pif4, pif5, pif3/4/5, pif1/3/4/5和pif1/3/4突變體并對突變效果進(jìn)行了驗(yàn)證。將突變體和野生型種子在含有ABA的培養(yǎng)基中培養(yǎng)烛谊，與野生型相比嘉汰，pif突變體在幼苗生長階段鞋怀，呈現(xiàn)出對ABA脫敏的表型。

3. 過表達(dá)PIF3和PIF5增強(qiáng)了幼苗對脫落酸的敏感性

為進(jìn)一步驗(yàn)證PIFs轉(zhuǎn)錄因子在ABA信號通路中的作用焙矛，將PIF3和PIF5的過表達(dá)株系在ABA的培養(yǎng)基中培養(yǎng)残腌，與野生型相比抛猫，過表達(dá)植株的生長受到明顯抑制，呈現(xiàn)出對ABA超敏感的表型闺金。

4. 通過DAP-seq分析在基因組水平上鑒定PIF3和PIF5轉(zhuǎn)錄因子在DNA上的結(jié)合位點(diǎn)

已經(jīng)有多項(xiàng)研究掖看，使用ChIP-seq面哥，對PIFs轉(zhuǎn)錄因子在基因組水平的結(jié)合位點(diǎn)進(jìn)行了鑒定尚卫，但是ChIP-seq方法受限于特異性抗體的質(zhì)量，并且操作復(fù)雜刹泄，實(shí)驗(yàn)難度大怎爵。本研究使用了一種新的方法：DAP-seq（DNA親和純化測序）對PIF3和PIF5在基因組上結(jié)合位點(diǎn)進(jìn)行了鑒定。最終鑒定出PIF的保守motif為G-box姆蘸，與已有的研究結(jié)果一致，并且鑒定出一批ABA響應(yīng)的基因是PIFs的下游靶基因狂秦，比如PYL3推捐，PYL6, PYL12, SnRK2.2, CPK4, CPK6, ABI5, ABF3和KIN1。2020年2月堪簿，在Molecular Plant期刊上皮壁，使用酵母單雜交、EMSA等技術(shù)甜孤，證明PIF1畏腕、PIP4和PIF5能夠結(jié)合到ABI5啟動子區(qū)的G-box上（Qi et al., 2020）。DAP-Seq的結(jié)果把夸，與此結(jié)果相互印證铭污。本研究證明了DAP-seq的可行性和有效性嘹狞。

討論

該研究通過DAP-seq鑒定出很多與ABA相關(guān)的PIFs靶基因磅网、參與ABA合成的基因通路以及ABA信號轉(zhuǎn)導(dǎo)通路中的組分。揭示了PIFs參與ABA信號轉(zhuǎn)導(dǎo)的機(jī)制并非簡單的單向調(diào)節(jié)涧偷，還存在反饋調(diào)控閉環(huán)燎潮。

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