Transgene-independent heredity of RdDM-mediated transcriptional gene silencing of endogenous genes in rice.
Transgene-independent heredity of RdDM-mediated transcriptional gene silencing of endogenous genes in rice.
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DOI：
10.1111/pbi.12934
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发表时间：
2018-12
影响因子：
13.8
通讯作者：
Takaiwa F
中科院分区：
文献类型：
--
作者：
Wakasa Y;Kawakatsu T;Harada T;Takaiwa F
为了诱导水稻（Oryza sativa L.）内源基因的转录基因沉默（TGS），我们表达了每个启动子区域的双链RNA，从而诱导RNA定向DNA甲基化（RdDM）。我们的目标是组成性表达的基因编码calnexin （CNX）、蛋白二硫异构酶（PDIL1‐1）和luminal binding protein (BiP1)；内质网应激诱导基因（OsbZIP50）；以及编码α -球蛋白（Glb‐1）和谷蛋白B4 （GluB4）的种子特异性表达基因。4个基因的TGS效率较高（CNX，再生植株的66.7%;OsBiP1, 67.4%; OsbZIP50, 63.4%; GluB4, 66.1%），而PDIL1‐1和Glb‐1 TGS系的效率较低（33.3%）和Glb‐1 TGS系（10.5%）。研究了部分TGS系的TGS遗传、启动子区甲基化水平和靶基因沉默的特异性。在CNX和OsbZIP50 TGS系的后代中，即使通过分离去除触发基因（T - DNA），靶基因的抑制作用仍得以保留（胚乳除外）。通过去甲基化处理恢复CNX的TGS，在CNX启动子−1 ~−250 bp区域检测到TGS和CHG甲基化水平在TGS和逆转系之间存在显著差异，表明TGS与启动子甲基化水平密切相关。当glutin多基因家族的GluB4基因被靶向时，TGS对靶基因表现出特异性抑制，而转录后基因沉默则相反。在此基础上，对RdDM技术在水稻育种新技术中的应用前景和有待解决的问题进行了展望。
To induce transcriptional gene silencing (TGS) of endogenous genes of rice (Oryza sativa L.), we expressed double‐strand RNA of each promoter region and thus induced RNA‐directed DNA methylation (RdDM). We targeted constitutively expressed genes encoding calnexin (CNX), protein disulphide isomerase (PDIL1‐1) and luminal binding protein (BiP1); an endoplasmic reticulum stress‐inducible gene (OsbZIP50); and genes with seed‐specific expression encoding α‐globulin (Glb‐1) and glutelin‐B4 (GluB4). TGS of four genes was obtained with high efficiency (CNX, 66.7% of regenerated plants; OsBiP1, 67.4%; OsbZIP50, 63.4%; GluB4, 66.1%), whereas the efficiency was lower for PDIL1‐1 (33.3%) and Glb‐1 TGS lines (10.5%). The heredity of TGS, methylation levels of promoter regions and specificity of silencing of the target gene were investigated in some of the TGS lines. In progeny of CNX and OsbZIP50 TGS lines, suppression of the target genes was preserved (except in the endosperm) even after the removal of trigger genes (T‐DNA) by segregation. TGS of CNX was reverted by demethylation treatment, and a significant difference in CG and CHG methylation levels in the −1 to −250 bp region of the CNX promoter was detected between the TGS and revertant lines, suggesting that TGS is closely related to the methylation levels of promoter. TGS exhibited specific suppression towards the target gene compared with post‐transcriptional gene silencing when GluB4 gene from glutelin multigene family was targeted. Based on these results, future perspectives and problems to be solved in the application of RdDM to new plant breeding techniques in rice are discussed.
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DOI：
10.1126/science.1224839
发表时间：
2012-09-14
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Ibarra CA;Feng X;Schoft VK;Hsieh TF;Uzawa R;Rodrigues JA;Zemach A;Chumak N;Machlicova A;Nishimura T;Rojas D;Fischer RL;Tamaru H;Zilberman D
通讯作者：
Zilberman D
通过苹果潜在球形病毒诱导的转录基因沉默的植物启动子序列中DNA甲基化的诱导和维持。
影响因子：
5.2
作者：
Kon T;Yoshikawa N
通讯作者：
Yoshikawa N
DOI：
10.1073/pnas.1413053112
发表时间：
2015-01-20
影响因子：
11.1
作者：
Bond, Donna M.;Baulcombe, David C.
通讯作者：
Baulcombe, David C.
影响因子：
56.9
作者：
Kinoshita, T;Miura, A;Kakutani, T
通讯作者：
Kakutani, T
影响因子：
5.8
作者：
Akman, Kemal;Haaf, Thomas;Tresch, Achim
通讯作者：
Tresch, Achim