水稻OsGRF6的克隆及调控初生根发育功能的初步分析
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1.厅市共建甘薯及特色豆科作物种质创新与利用四川省重点实验室;2.细胞活动与逆境适应教育部重点实验室;3.重庆三峡农业科学院

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甘肃省科技厅自然科学基金面上项目(21JR7RA484);厅市共建甘薯及特色豆科作物种质创新与利用四川省重点实验室开放课题基金资助项目(2021CGIHL04);河南大学作物逆境适应与改良国家重点实验室开放课题(2022KF04)


Cloning and a preliminary functional analysis of OsGRF6 on regulating primary root development in rice
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    摘要:

    OsGRF6隶属于GRF家族,参与调控植物的生长发育和调控植物对非生物胁迫的耐受性。为了研究OsGRF6调控水稻初生根发育的分子机制,该研究从水稻品种‘ZH11’中克隆得到转录因子基因OsGRF6,并对其进行了进化树构建,启动子分析,并对OsGRF6-OE和osgrf6进行了基因型分析和qRT-PCR鉴定及水稻表型观察,并通过酵母单杂交检测了OsGRF6的结合基序,进一步通过ChIP-seq和RNA-seq数据分析了OsGRF6下游的候选靶基因。结果显示:(1)OsGRF6含有一个QLQ和一个WRC保守的功能结构域,与拟南芥AtGRF1和AtGRF2以及水稻OsGRF7亲缘关系较近。OsGRF6启动子区域含有多个非生物胁迫和激素响应顺式作用元件。(2)OsGRF6在水稻种子、幼穗和根中表达量较高,而在叶中表达量较低。亚细胞定位显示,OsGRF6定位于细胞核,并且OsGRF6具有转录激活活性。(3)与野生型相比,过表达OsGRF6的转基因材料表现出初生根长度较长,而突变体材料则表现出初生根变短的表型。(4)OsGRF6能与CGGCA基序结合。(5)结合ChIP-seqs和RNA-seqs分析发现,OsGRF6靶基因中包含多个参与调控水稻根发育的基因,如OsARF7,OsARF4等。

    Abstract:

    OsGRF6 belongs to the GRF family and is involved in the regulation of plant growth and development and tolerance to abiotic stress. In order to study the molecular mechanism of OsGRF6 to regulate the primary root development of rice, the transcription factor gene OsGRF6 was cloned from the rice variety ''ZH11''. The evolutionary tree construction and promoter analysis were carried out. The genotype analysis, qRT-PCR identification and rice phenotype observation of OsGRF6-OE and osgrf6 were carried out. The binding sequence of OsGRF6 was detected through yeast one-hybrid assays, and the candidate target genes downstream of OsGRF6 were further analyzed through ChIP-seqs and RNA-seqs data. The results show that: (1) OsGRF6 contains a QLQ and a WRC conservative functional domain, which is closely related to AtGRF1 and AtGRF2 and OsGRF7. The OsGRF6 promoter region contains multiple abiotic stress and hormone response cis-action elements. (2) OsGRF6 showed a higher level of expression in rice seeds, young spikes and roots than in leaf. Subcellular localization shows that OsGRF6 is located in the nucleus, and OsGRF6 has transcription activation activity. (3) Compared with wild types, overexpression of OsGRF6 transgenic plant materials show a longer primary root length, while loss of function mutant osgrf6 show a shorter phenotype of primary roots. (4) OsGRF6 can bind to CGGCA sequence. (5) Combined with ChIP-seqs and RNA-seqs analysis, it is found that a number of the OsGRF6 target genes were involved in regulating rice root development, such as OsARF7 and OsARF4, etc.

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余青青,单玉爽,尚汇鑫,等.水稻OsGRF6的克隆及调控初生根发育功能的初步分析[J].西北植物学报,2024,44(1):22-33

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  • 收稿日期:2023-05-05
  • 最后修改日期:2023-10-24
  • 录用日期:2023-11-02
  • 在线发布日期: 2024-01-02
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