[1]宋刚,方志刚,王玉龙,等.柳枝稷BiP基因的鉴定、表达分析和抗逆境功能[J].江苏农业学报,2022,38(04):889-899.[doi:doi:10.3969/j.issn.1000-4440.2022.04.004]
　SONG Gang,FANG Zhi-gang,WANG Yu-long,et al.Identification, expression analysis and function in adverse environment of BiP in switchgrass[J].,2022,38(04):889-899.[doi:doi:10.3969/j.issn.1000-4440.2022.04.004]
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柳枝稷BiP基因的鉴定、表达分析和抗逆境功能()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年04期
页码:
889-899
栏目:
遗传育种·生理生化
出版日期:
2022-08-31

文章信息/Info

Title:
Identification, expression analysis and function in adverse environment of BiP in switchgrass
作者:
宋刚1; 2; 方志刚3; 王玉龙2; 蔡庆生2
(1.江苏农林职业技术学院茶与食品科技学院，江苏句容212400；2.南京农业大学生命科学学院，江苏南京210095；3.喀什大学生命与地理科学学院，新疆喀什844099）
Author(s):
SONG Gang1; 2; FANG Zhi-gang3; WANG Yu-long2; CAI Qing-sheng2
(1.College of Tea and Food Science and Technology，Jiangsu Vocational College of Agriculture and Forestry, Jurong 212400, China;2.College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China;3.College of Life and Geographic Sciences, Kashi University, Kashi 844006, China)
关键词:
柳枝稷; BiP; 表达分析; 胁迫; 酵母转化
Keywords:
switchgrass; BiP; expression analysis; stress; yeast transformation
分类号:
Q786
DOI:
doi:10.3969/j.issn.1000-4440.2022.04.004
文献标志码:
A
摘要:
有害环境条件对植物生长发育有不利影响，并会引起蛋白质变性或错误折叠。分子伴侣之一的绑定蛋白（BiP）通过减轻错误折叠蛋白质引起的内质网应激发挥重要的保护作用。柳枝稷（Panicum virgatum L.）是一种原产于北美、多年生C4型禾本科植物。由于其生物量大、适应性强，近年来被作为能源植物研究和利用。为明确柳枝稷中BiP基因及其在逆境下的表达和功能，本研究通过对柳枝稷基因组数据本地BLAST比对，结合具有BiP保守结构域和内质网驻留信号序列（HDEL）特征筛选的条件，共在柳枝稷基因组中鉴定出4个BiP基因，命名为PvBiP1a、PvBiP1b、PvBiP2、PvBiP3，分别位于1号和5号染色体上。多物种序列比对和进化分析结果表明，PvBiPs具有BiP蛋白典型功能结构域，PvBiP1a和PvBiP1b聚成一个进化枝，并与水稻OsBiP1聚为一类，都含有7个内含子结构；PvBiP2与PvBiP3聚为另一进化枝，无内含子结构。上游2 kb启动子序列分析结果表明，PvBiPs主要包括胁迫防御和激素应答顺式作用元件。PvBiPs受内质网胁迫诱导剂二硫苏糖醇诱导表达，在NaCl、PEG、ABA和CdCl2胁迫下表现出上调表达，PvBiP2相对表达量的峰值最高。将PvBiP1a、PvBiP2与PvBiP3克隆至pGAD426载体并转化酵母，在CdCl2、NaCl和甘露醇胁迫下进行耐性验证，结果显示，3个基因对镉和盐胁迫都有耐性，对甘露醇胁迫无耐性。本研究结果将为深入挖掘柳枝稷BiP基因功能、提升柳枝稷在逆境下尤其是对重金属镉胁迫的耐性提供理论基础。
Abstract:
Adverse environmental conditions have detrimental impact on plant growth and development, and can cause protein denaturation or misfolding. As one of molecular chaperones, the binding protein (BiP) plays an important protective effect by alleviating endoplasmic reticulum (ER) stress induced by misfolded proteins. Switchgrass (Panicum virgatum L.) is a kind of perennial C4 plant of grass family originated from North America, it has been studied and utilized as one of the energy crops in recent years due to the characteristics of high biomass yield and strong adaptability. To identify BiP genes of switchgrass and their expression and function in adverse environment, local BLAST of switchgrass genomic data was carried out, combined with screening conditions with characteristics of BiP conserved domain and ER retention signal sequence HDEL, four BiP genes named PvBiP1a, PvBiP1b, PvBiP2 and PvBiP3 were identified from the genome of switchgrass, which located on chromosome 1 and 5, respectively. Results of multi-species sequence analysis and evolutionary analysis showed that, PvBiPs had the typical functional domain of BiP, PvBiP1a and PvBiP1b clustered into a clade, and clustered into a category with rice OsBiP1, both PvBiP1a and PvBiP1b contained seven introns. PvBiP2 and PvBiP3 clustered into another clade, and neither of them had intron. Analysis results of upstream 2 kb promoter sequences showed that, the PvBiPs mainly consisted of cis-elements of stress defense and hormone response. Expressions of PvBiPs were induced by ER stress inducer dithiothreitol and were up-regulated under NaCl, polyethylene glycol, ABA and CdCl2 stresses. Among them, PvBiP2 had the highest peak value of relative expression level. PvBiP1a, PvBiP2 and PvBiP3 were then cloned into pGAD426 vector and the vector was used to transform yeast to verify the tolerance under CdCl2, NaCl and mannitol stresses. The results showed that, PvBiPs were tolerant to Cd stress and salt stress, but were not tolerant to mannitol stress. The results of this study can provide theoretical base for further exploring of the function of BiP gene in switchgrass and improvement of tolerance to adverse situation (especially under Cd stress) of switchgrass.

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备注/Memo

备注/Memo:
收稿日期：2021-12-16基金项目：国家自然科学基金面上项目（31372359）；江苏省教育厅面上项目（19KB180015）作者简介：宋刚（1978-），男，江苏盐城人，博士，副教授，主要从事植物环境生理和快繁技术的研究。（E-mail）songgang@jsafc.edu.cn通讯作者：蔡庆生，（Tel）025-84395187；（E-mail）qscai@njau.edu.cn
更新日期/Last Update: 2022-09-06