Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing.
Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing.
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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。
DOI：
10.3389/fmicb.2014.00595
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发表时间：
2014
影响因子：
5.2
通讯作者：
Yoshikawa N
中科院分区：
文献类型：
--
作者：
Kon T;Yoshikawa N
苹果潜伏球形病毒（ALSV）是一种高效的病毒诱导的基因沉默载体，可用于植物功能基因组学研究。在这里，农杆菌介导的接种（农杆菌接种）系统开发的ALSV载体，和病毒诱导的转录基因沉默（VITGS）被描述在植物感染的ALSV载体。将ALSV RNA 1和RNA 2的cDNA分别插入到花椰菜花叶病毒35 S启动子和NOS-T序列之间，构建成双元载体pCAMBIA 1300，分别构建成pCALSR 1和pCALSR 2-XSB或pCALSR 2-XSB/MN。当将这些载体构建体农杆菌接种到具有表达病毒沉默抑制子的构建体的本氏烟草植物中时，载体的感染效率为100%。用带有部分35 S启动子序列的重组ALSV载体在一株烟草中诱导绿色荧光蛋白基因的转录沉默。benthamiana植物，导致受感染植物的绿色荧光消失。亚硫酸氢盐测序表明，在CG和CHG位点的35 S启动子序列的胞嘧啶残基高度甲基化的沉默的零代植物感染的ALSV携带的启动子序列，以及在后代。ALSV介导的VITGS状态由后代遗传多代。此外，诱导VITGS的内源基因（查耳酮脱氢酶-A）被证明在矮牵牛植物感染的ALSV载体携带的天然启动子序列。这些结果表明，基于ALSV的载体可用于研究植物基因组中的DNA甲基化，并通过表观遗传修饰为植物育种提供了有用的工具。
Apple latent spherical virus (ALSV) is an efficient virus-induced gene silencing vector in functional genomics analyses of a broad range of plant species. Here, an Agrobacterium-mediated inoculation (agroinoculation) system was developed for the ALSV vector, and virus-induced transcriptional gene silencing (VITGS) is described in plants infected with the ALSV vector. The cDNAs of ALSV RNA1 and RNA2 were inserted between the cauliflower mosaic virus 35S promoter and the NOS-T sequences in a binary vector pCAMBIA1300 to produce pCALSR1 and pCALSR2-XSB or pCALSR2-XSB/MN. When these vector constructs were agroinoculated into Nicotiana benthamiana plants with a construct expressing a viral silencing suppressor, the infection efficiency of the vectors was 100%. A recombinant ALSV vector carrying part of the 35S promoter sequence induced transcriptional gene silencing of the green fluorescent protein gene in a line of N. benthamiana plants, resulting in the disappearance of green fluorescence of infected plants. Bisulfite sequencing showed that cytosine residues at CG and CHG sites of the 35S promoter sequence were highly methylated in the silenced generation zero plants infected with the ALSV carrying the promoter sequence as well as in progeny. The ALSV-mediated VITGS state was inherited by progeny for multiple generations. In addition, induction of VITGS of an endogenous gene (chalcone synthase-A) was demonstrated in petunia plants infected with an ALSV vector carrying the native promoter sequence. These results suggest that ALSV-based vectors can be applied to study DNA methylation in plant genomes, and provide a useful tool for plant breeding via epigenetic modification.
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影响因子：
4.5
作者：
Liu S;Yeh CT;Ji T;Ying K;Wu H;Tang HM;Fu Y;Nettleton D;Schnable PS
通讯作者：
Schnable PS
影响因子：
7.2
作者：
Brigneti, G;Martín-Hernández, AM;Jones, JDG
通讯作者：
Jones, JDG
影响因子：
7.2
作者：
BAULCOMBE, DC;CHAPMAN, S;CRUZ, SS
通讯作者：
CRUZ, SS
影响因子：
56.9
作者：
BENFEY, PN;CHUA, NH
通讯作者：
CHUA, NH
DOI：
10.1038/nrg2719
发表时间：
2010-03
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
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