前期研究成果：
何祖華研究團隊與育種家合作，從2002開始，廣泛篩選抗瘟種質(zhì)，在起源于我國農(nóng)家品種的育種材料（GM4）中成功鑒定出持久廣譜抗稻瘟病基因pigm，并揭示了水稻廣譜抗病與產(chǎn)量平衡的表觀調(diào)控新機制。
summary:
1.the rice Pigm locus contains a cluster of genes encoding nucleotide-binding leucine-rich repeat (NLR) receptors that confer durable resistance to the fungus Magnaporthe oryzae without yield penalty.
2.Among these NLR receptors, PigmR confers broad-spectrum resistance, whereas PigmS competitively attenuates PigmR homodimerization to suppress resistance.
3.PigmS expression, and thus PigmR-mediated resistance, are subjected to tight epigenetic regulation. PigmS increases seed production to counteract the yield cost induced by PigmR.
4. our study reveals a mechanism balancing high disease resistance and yield through epigenetic regulation of paired antagonistic NLR receptors, providing a tool to develop elite crop varieties.
待解決問題：
the molecular mechanisms underlying Pigm R mediated broad spectrum resistance remained elusive。
新發(fā)現(xiàn)：summary
1.we uncover a previously unappreciated protein binding partner of Pigm R, PIBP1, an RNA recognition motif (RRM)-containing protein.
2.We show that PIBP1 and its homolog,Os06 g02240, are speci?cally required to mediate resistance by Pigm R and other broad-spectrum NLRs, but not NLRs with limited race-speci?c resistance.
3.We provide evidence that PIBP1 and Os06 g02240 are unconventional transcription factors that directly activate the expression of the defense genes Os WAK14 and Os PAL1.
4.Thus, our work identi?es previously unappreciated regulators of ETI and RRM-containing transcription factors that regulate immunity in plants.
感興趣的部分實驗結(jié)果/陰性對照設置/原生質(zhì)體觀察及統(tǒng)計學分析可參考。
Q1：punch（使用稻瘟菌TH12） 實驗,為何使用fungal 28SrDNA檢測 lesion的relative growth? 與使用稻瘟菌的maker基因檢測有何不同？我想想......
Q2：關(guān)于PIBP1核定位觀察及pigmR/S對其在細胞核中的積累變化，使用共聚焦顯微觀察100以上的細胞后做統(tǒng)計學分析。我們可以看出pigmS明顯減弱PIBP1在nulear的表達，但pigmR和PIBP1（S275A）點突對PIBP1的nulear定位位圖，我肉眼看去和CK差異不明顯，雖統(tǒng)計學數(shù)據(jù)能表示出明顯差異。畢竟顯微觀察多少有主觀意識判斷差異，是否可以通過轉(zhuǎn)原生質(zhì)體/NB后提核蛋白跑western 檢測（上樣定量）差異。
Q3：PIBP1-OE構(gòu)建（使用PUN1301-PUbi-GFP vector，轉(zhuǎn)農(nóng)桿（EHA105，RT/western blot檢測），與native promoter +genomic序列構(gòu)建自身啟動子的相比，哪個更好？
