百脉根中Lj5NG4与结瘤因子受体LjNFR1互作调控根瘤菌侵染

doi:10.14088/j.cnki.issn0439-8114.2020.05.029

湖北农业科学 ›› 2020, Vol. 59 ›› Issue (5): 145-151.doi: 10.14088/j.cnki.issn0439-8114.2020.05.029

百脉根中Lj5NG4与结瘤因子受体LjNFR1互作调控根瘤菌侵染

宋嘉宾, 端木德强

华中农业大学生命科学技术学院,武汉 430070

修回日期:2019-05-13 出版日期:2020-03-10 发布日期:2020-06-05
作者简介:宋嘉宾（1993-）,女,山东烟台人,硕士,研究方向为微生物,（电话）18202764573（电子信箱）644290533@qq.com。

The interaction of Lj5NG4 and LjNFR1 in Lotus japonicus regulates the infection of Rhizobium

SONG Jia-bin, DUANMU De-qiang

College of Life Science and Technology of Huazhong Agricultural University,Wuhan 430070,China

Revised:2019-05-13 Online:2020-03-10 Published:2020-06-05

摘要/Abstract

摘要： 以百脉根结瘤因子受体激酶LjNFR1为诱饵,通过建立百脉根根瘤共生不同时期的酵母膜双杂文库筛选获得了一个与之互作的生长素相关转运蛋白Lj5NG4。通过酵母双杂交和烟草Split-Luc等试验进一步证实了Lj5NG4与LjNFR1的蛋白互作。利用CRISPR-Cas9基因敲除及超表达技术创建相关转基因材料,研究发现Lj5NG4可能在根瘤菌侵染前期起负调控作用,但对结瘤过程没有显著影响。同时启动子表达也证实了Lj5NG4在侵染前期根毛中强烈表达,成熟根瘤内部不表达。外源施加生长素试验发现低浓度的生长素会减弱Lj5NG4突变体的早期侵染表型,这表明生长素在共生结瘤发育中通过Lj5NG4发挥功能。

关键词: 百脉根, 根瘤菌侵染, LjNFR1, Lj5NG4, 生长素

Abstract: ： Using the Lotus japonicus nodule factor receptor kinase LjNFR1 as a bait and a yeast membrane two hybrid library constructed with transcripts harvested at different stages of nodule symbiosis, we identified an auxin related transporter protein Lj5NG4. The interaction between Lj5NG4 and LjNFR1 was further confirmed by yeast two hybrid and Split-Luc experiments. Using CRISPR-Cas9 and overexpression technology, we conclude that Lj5NG4 may play a negative regulatory role in the early stage of rhizobial infection, but had no significant effect on nodule formation. Meanwhile, promoter expression also confirmed that Lj5NG4 was strongly expressed in root hairs at the early stage of infection, but not in inner mature nodules. Exogenously applied auxin treatment showed that low concentration of auxin weakened the phenotype of Lj5NG4 mutant, indicating that auxin plays a role in the development of symbiotic nodules.

Key words: Lotus japonicus, rhizobium infection, LjNFR1, Lj5NG4, auxin

中图分类号:

Q789

宋嘉宾, 端木德强. 百脉根中Lj5NG4与结瘤因子受体LjNFR1互作调控根瘤菌侵染[J]. 湖北农业科学, 2020, 59(5): 145-151.

SONG Jia-bin, DUANMU De-qiang. The interaction of Lj5NG4 and LjNFR1 in Lotus japonicus regulates the infection of Rhizobium[J]. HUBEI AGRICULTURAL SCIENCES, 2020, 59(5): 145-151.

参考文献

[1] RADUTOIU S,MADSEN L,MADSEN E,et al.Plant recognition of symbiotic bacteria requires two LysM receptor-like kinases[J].Nature,2003,425:585-592.
[2] MADSEN E,MADSEN L,RADUTOIU S,et al.A receptor kinase gene of the LysM type is involved in legume perception of rhizobial signals[J].Nature,2003,425:637-640.
[3] LIMPENS E,FRANKEN C,SMIT P,et al.LysM domain receptor kinases regulating rhizobial Nod factor-induced infection[J].Science,2003,302:630-633.
[4] SMIT P,LIMPENS E,GEURTS R,et al.Medicago LYK3,an entry receptor in rhizobial nodulation factor signaling[J].Plant physiology,2007,145:183-191.
[5] MULDER L,LEFEBVRE B,CULLIMORE J,et al.LysM domains of Medicago truncatula NFP protein involved in Nod factor perception. Glycosylation state, molecular modeling and docking of chitooligosaccharides and Nod factors[J].Glycobiology,2006,16:801-809.
[6] OLDROYD G,MURRAY J,POOLE P,et al.The rules of engagement in the legume-rhizobial symbiosis[J].Annual review of genetics,2011,45:119-144.
[7] MADSEN E,ANTOLÍN-LLOVERA M,GROSSMANN C,et al. Autophosphorylation is essential for the in vivo function of the Lotus japonicus Nod factor receptor 1 and receptor-mediated signaling in cooperation with Nod factor receptor 5[J].The plant journal,2011,65:404-417.
[8] MBENGUE M,CAMUT S,CARVALHO-NIEBEL F,et al.The Medicago truncatula E3 ubiquitin ligase PUB1 interacts with the LYK3 symbiotic receptor and negatively regulates infection and nodulation[J].Plant cell,2010,22:3474-3488.
[9] LEFEBVRE B,TIMMERS T,MBENGUE M,et al.A remorin protein interacts with symbiotic receptors and regulates bacterial infection[J].Proc Natl Acad Sci USA.,2010,107(5):2343-2350.
[10] PHILIPPE R,OANA D,REKA NA,et al.Arabidopsis WAT1 is a vacuolar auxin transport facilitator required for auxin homoeostasis[J].Nature communations,2013,4:25-26.
[11] CHAPMAN E J,ESTELLE M.Mechanism of auxin-regulated gene expression in plants[J].Annu Rev Genet,2009,43:265-285.
[12] BREAKSPEAR A,LIU C,ROY S,et al.The root hair "infectome" of Medicago truncatula uncovers changes in cell cycle genes and reveals a requirement for Auxin signaling in rhizobial infection[J].Plant cell,2014,26:4680-4701.
[13] ROY S,ROBSON F,LILLEY J,et al.MtLAX2,a functional homologue of the Arabidopsis auxin influx transporter AUX1,is required for nodule organogenesis[J].Plant Physiol,2017,174:326-338.
[14] HUO X,SCHNABEL E,HUGHES K,et al.RNAi phenotypes and the localization of a protein:GUS fusion imply a role for Medicago truncatula PIN genes in nodulation[J].J Plant growth regul,2006,25:156-165.
[15] NICOLAS D,BORIS S,LAURENT D.Emerging functions of nodulin-like proteins in non-nodulating plant species[J].Plant cell physiol,2014,55(3):469-474.
[16] PHILIPPE R,NICOLAS D,RUBEN V,et al.Walls are thin 1 (WAT1), an Arabidopsis homolog of Medicago truncatula NODULIN21,is a tonoplast-localized protein required for secondary wall formation in fibers[J].The plant journal,2010,63:469-483.

百脉根中Lj5NG4与结瘤因子受体LjNFR1互作调控根瘤菌侵染

The interaction of Lj5NG4 and LjNFR1 in Lotus japonicus regulates the infection of Rhizobium

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

[1]	杨统一, 杜秋霞, 刘静霏, 翟德丽, 刘延鹏, 赵卫国. 一株桑树根际促生菌的筛选鉴定及促生性能研究[J]. 湖北农业科学, 2021, 60(1): 80-84.
[2]	张信玲, 李新艺, 李娜, 孔令芳. 凸尖杜鹃组培快繁体系的构建[J]. 湖北农业科学, 2019, 58(9): 124-127.