Rootstock-to-scion transfer of transgene-derived small interfering RNAs and their effect on virus resistance in nontransgenic sweet cherry.
Rootstock-to-scion transfer of transgene-derived small interfering RNAs and their effect on virus resistance in nontransgenic sweet cherry.
复制标题
DOI：
10.1111/pbi.12243
复制
发表时间：
2014-12
影响因子：
13.8
通讯作者：
Dongyan Zhao;G. Song
中科院分区：
文献类型：
--
作者：
Dongyan Zhao;G. Song
小干扰 RNA (siRNA) 是植物中的沉默信号。通过siRNA介导的基因沉默开发的抗病毒转基因砧木可以通过从转基因砧木转运的siRNA来增强非转基因接穗的病毒抗性。然而，木本植物中外源基因的siRNA从砧木到接穗移动的令人信服的证据仍然缺乏。为了确定是否可以转移外源 siRNA，将非转基因甜樱桃（接穗）嫁接到转基因樱桃砧木（TR）上，该砧木用 RNA 干扰（RNAi）载体转化，表达李属坏死环斑病毒（PNRSV-hpRNA）基因组 RNA3 的短发夹 RNA。使用位于嫁接接头上方约 1.2 m 的 TR 和嫁接树（砧木/接穗）的芽组织进行小 RNA 测序。 siRNA图谱的比较表明PNRSV-hpRNA能够有效地产生siRNA并消除TR中的PNRSV。此外，我们的研究首次证实，PNRSV-hpRNA 衍生的 siRNA 从转基因砧木长距离（1.2 m）转移到木本植物中的非转基因接穗。用PNRSV接种非转基因接穗表明，转移的siRNA增强了嫁接到TR上的接穗的PNRSV抗性。总的来说，这些发现为“使用转基因砧木在果树中生产非转基因接穗产品”提供了基础。
Small interfering RNAs (siRNAs) are silencing signals in plants. Virus-resistant transgenic rootstocks developed through siRNA-mediated gene silencing may enhance virus resistance of nontransgenic scions via siRNAs transported from the transgenic rootstocks. However, convincing evidence of rootstock-to-scion movement of siRNAs of exogenous genes in woody plants is still lacking. To determine whether exogenous siRNAs can be transferred, nontransgenic sweet cherry (scions) was grafted on transgenic cherry rootstocks (TRs), which was transformed with an RNA interference (RNAi) vector expressing short hairpin RNAs of the genomic RNA3 of Prunus necrotic ringspot virus (PNRSV-hpRNA). Small RNA sequencing was conducted using bud tissues of TRs and those of grafted (rootstock/scion) trees, locating at about 1.2 m above the graft unions. Comparison of the siRNA profiles revealed that the PNRSV-hpRNA was efficient in producing siRNAs and eliminating PNRSV in the TRs. Furthermore, our study confirmed, for the first time, the long-distance (1.2 m) transfer of PNRSV-hpRNA-derived siRNAs from the transgenic rootstock to the nontransgenic scion in woody plants. Inoculation of nontransgenic scions with PNRSV revealed that the transferred siRNAs enhanced PNRSV resistance of the scions grafted on the TRs. Collectively, these findings provide the foundation for 'using transgenic rootstocks to produce products of nontransgenic scions in fruit trees'.