甘蔗类钙调素ScCML13与SCMV运动蛋白P3N-PIPO的互作研究

类钙调素(Calmodulin-like,CML)是植物特有的钙信号感受器蛋白之一,参与植物生长发育和响应外界环境信号转导.甘蔗(Saccharum spp.hybrid)中CML应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)侵染尚未见报道.本研究从甘蔗热带种 Badila(S.officinarum)中克隆了一个 CML 基因,命名为 ScCML13.该基因开放读码框(open reading frame,ORF)长度为 519 bp,编码 172 aa.生物信息学分析表明,ScCML13 属于稳定的亲水脂溶蛋白,无跨膜结构域,有 4 个结合 Ca2+的 EF-h...

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Published in作物学报 Vol. 50; no. 7; pp. 1855 - 1866
Main Authors 玉泉馨, 杨宗桃, 张海, 程光远, 焦文迪, 曾康, 罗廷绪, 黄国强, 王璐, 徐景升
Format Journal Article
LanguageChinese
Published 福建农林大学/农业农村部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗工程技术研究中心/教育部作物遗传育种与综合利用重点实验室,福建福州 350002 12.07.2024
Subjects
甘蔗花叶病毒
蛋白互作
Sugarcane mosaic virus
protein interaction
calmodulin-like protein
类钙调素
P3N-PIPO
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ISSN0496-3490
DOI10.3724/SP.J.1006.2024.34191

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Abstract 类钙调素(Calmodulin-like,CML)是植物特有的钙信号感受器蛋白之一,参与植物生长发育和响应外界环境信号转导.甘蔗(Saccharum spp.hybrid)中CML应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)侵染尚未见报道.本研究从甘蔗热带种 Badila(S.officinarum)中克隆了一个 CML 基因,命名为 ScCML13.该基因开放读码框(open reading frame,ORF)长度为 519 bp,编码 172 aa.生物信息学分析表明,ScCML13 属于稳定的亲水脂溶蛋白,无跨膜结构域,有 4 个结合 Ca2+的 EF-hand结构域;系统进化树分析表明,该蛋白在单子叶和双子叶植物中及单子叶 C3 和C4 植物中具有明显分化;酵母双杂交(yeast two-hybrid,Y2H)和双分子荧光互补(bimolecular fluorescence complemen-tation,BiFC)试验表明,ScCML13 与SCMV的运动蛋白P3N-PIPO互作.亚细胞定位试验表明ScCML13 定位于内质网和细胞核.共定位试验发现ScCML13会干扰SCMV-P3N-PIPO的质膜(plasma membrane)或胞间连丝(plasmodesmata)定位.实时荧光定量PCR(Real Time Quantitative PCR,RT-qPCR)分析发现,ScCML13 基因主要在甘蔗叶片中表达,在节间和根中的相对表达量低;ScCML13基因在感染SCMV后2 h显著上调,随后下调至与对照组相比的水平,而在感染后期上调.
AbstractList 类钙调素(Calmodulin-like,CML)是植物特有的钙信号感受器蛋白之一,参与植物生长发育和响应外界环境信号转导.甘蔗(Saccharum spp.hybrid)中CML应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)侵染尚未见报道.本研究从甘蔗热带种 Badila(S.officinarum)中克隆了一个 CML 基因,命名为 ScCML13.该基因开放读码框(open reading frame,ORF)长度为 519 bp,编码 172 aa.生物信息学分析表明,ScCML13 属于稳定的亲水脂溶蛋白,无跨膜结构域,有 4 个结合 Ca2+的 EF-hand结构域;系统进化树分析表明,该蛋白在单子叶和双子叶植物中及单子叶 C3 和C4 植物中具有明显分化;酵母双杂交(yeast two-hybrid,Y2H)和双分子荧光互补(bimolecular fluorescence complemen-tation,BiFC)试验表明,ScCML13 与SCMV的运动蛋白P3N-PIPO互作.亚细胞定位试验表明ScCML13 定位于内质网和细胞核.共定位试验发现ScCML13会干扰SCMV-P3N-PIPO的质膜(plasma membrane)或胞间连丝(plasmodesmata)定位.实时荧光定量PCR(Real Time Quantitative PCR,RT-qPCR)分析发现,ScCML13 基因主要在甘蔗叶片中表达,在节间和根中的相对表达量低;ScCML13基因在感染SCMV后2 h显著上调,随后下调至与对照组相比的水平,而在感染后期上调.
Abstract_FL Calmodulin-like(CML),one of the calcium signal receptor proteins unique to plants,is involved in plant growth and development and in response to environmental signal transduction.However,the response of CML to Sugarcane mosaic virus(SCMV)infection in sugarcane(Saccharum spp.hybrid)has not been reported.In the present study,a CML gene was cloned from a Badila(S.officinarum)and designated as ScCML13.The open reading frame(ORF)of ScCML13 gene is 519 bp in length and encodes a protein with 172 aa in length.Bioinformatics analysis showed that ScCML13 was a stable hydrophilic lipoprotein in with no transmembrane domain and contains 4 Ca2+-binding EF-hand domains.Phylogenetic tree analysis showed that the ScCML13 protein differentiated in monocotyledon and dicotyledon plants and in monocotyledonous C3 and C4 plants.Yeast two-hybrid(Y2H)and bimolecular fluorescence complementation(BiFC)experiments indicated that ScCML13 interacted with P3N-PIPO,the movement protein of SCMV.Subcellular localization assays demonstrated that ScCML13 was localized to the endoplasmic reticulum and nucleus,whereas the co-localization assays showed that ScCML13 interfered with the localization of SCMV-P3N-PIPO to plasma membrane or plasmodesmata.The RT-qPCR showed that ScCML13 gene was mainly expressed in sugarcane leaves,but relatively low in internodes and roots.The ScCML13 gene was significantly up-regulated at 2 h upon SCMV infection,and then down-regulated to the levels compared with the control group,while up-regulated at the later stage of infection.
Author 罗廷绪
曾康
玉泉馨
程光远
杨宗桃
王璐
徐景升
张海
焦文迪
黄国强
AuthorAffiliation 福建农林大学/农业农村部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗工程技术研究中心/教育部作物遗传育种与综合利用重点实验室,福建福州 350002
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Author_FL HUANG Guo-Qiang
YU Quan-Xin
LUO Ting-Xu
WANG Lu
YANG Zong-Tao
ZHANG Hai
CHENG Guang-Yuan
JIAO Wen-Di
ZENG Kang
XU Jing-Sheng
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DocumentTitle_FL Interaction between calmodulin-like ScCML13 of sugarcane and SCMV move-ment protein P3N-PIPO
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Keywords 甘蔗花叶病毒
蛋白互作
Sugarcane mosaic virus
protein interaction
calmodulin-like protein
类钙调素
P3N-PIPO
Language Chinese
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Snippet 类钙调素(Calmodulin-like,CML)是植物特有的钙信号感受器蛋白之一,参与植物生长发育和响应外界环境信号转导.甘蔗(Saccharum spp.hybrid)中CML应答甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)侵染尚未见报道.本研究从甘蔗热带种...
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Title 甘蔗类钙调素ScCML13与SCMV运动蛋白P3N-PIPO的互作研究
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