中国农业科学 ›› 2020, Vol. 53 ›› Issue (17): 3432-3442.doi: 10.3864/j.issn.0578-1752.2020.17.002
收稿日期:
2019-12-30
接受日期:
2020-03-08
出版日期:
2020-09-01
发布日期:
2020-09-11
通讯作者:
游均
作者简介:
刘爱丽,E-mail:基金资助:
LIU AiLi(),WEI MengYuan,LI DongHua,ZHOU Rong,ZHANG XiuRong,YOU Jun()
Received:
2019-12-30
Accepted:
2020-03-08
Online:
2020-09-01
Published:
2020-09-11
Contact:
Jun YOU
摘要:
【目的】SiGolS6是芝麻肌醇半乳糖苷合成酶家族成员,在植物抗旱过程中可能发挥重要作用。研究SiGolS6在植物干旱胁迫抗性中的功能,以期为芝麻抗旱遗传改良提供理论基础和基因资源。【方法】利用反转录PCR(reverse transcription PCR,RT-PCR)方法从芝麻中克隆获得肌醇半乳糖苷合成酶基因SiGolS6。使用InterProScan、ClustalX2和MEGA5.2等生物信息学软件对序列进行分析。通过对转SiGolS6拟南芥材料的表型分析和生理指标测定,研究SiGolS6在植物抗旱性中的功能及作用机理。【结果】克隆获得SiGolS6的全长CDS序列为921 bp,编码306个氨基酸,其蛋白质分子量为35.07 kD,等电点为4.75。序列分析显示SiGolS6蛋白含有糖基转移酶保守结构域(IPR002495),属于糖基转移酶超家族。与其他物种GolS蛋白构建的系统进化树中,SiGolS6与马铃薯中的同源基因相似度较高。利用潮霉素筛选和PCR鉴定,获得6个独立的转基因拟南芥株系;qRT-PCR分析筛选出表达量较高的3个转基因株系(OE-2、OE-3和OE-4)用于后续试验分析,检测发现这三个转基因株系的棉子糖含量显著高于野生型植株。干旱胁迫下,转基因株系萎蔫程度较野生型轻,干旱胁迫28 d和复水恢复生长5 d后,转基因株系的鲜重均显著高于野生型,而正常供水条件下无明显差异;干旱胁迫28 d后复水生长5 d,50%的转基因植株基本恢复,而野生型植株存活率不到10%。干旱胁迫21 d,转基因植株的相对电导率,活性氧积累和相对MDA含量均显著低于野生型,而相对SOD、POD活性则显著高于野生型。【结论】在拟南芥中超量表达芝麻SiGolS6,能够提高植物在干旱胁迫中的耐受性。
刘爱丽,魏梦园,黎冬华,周瑢,张秀荣,游均. 芝麻肌醇半乳糖苷合成酶基因SiGolS6的克隆及功能分析[J]. 中国农业科学, 2020, 53(17): 3432-3442.
LIU AiLi,WEI MengYuan,LI DongHua,ZHOU Rong,ZHANG XiuRong,YOU Jun. Cloning and Function Analysis of Sesame Galactinol Synthase Gene SiGolS6 in Arabidopsis[J]. Scientia Agricultura Sinica, 2020, 53(17): 3432-3442.
图1
SiGolS6和其他物种GolS的多序列比对(A)和系统进化分析(B) SiGolS6:芝麻Sesamum indicum,XP_011087512.1;CsGolS:甜橙Citrus sinensis,XP_006444560.1;PtGolS:毛果杨Populus trichocarpa,XP_011087512.1;AtGolS1:拟南芥Arabidopsis thaliana,NP_182240.1;GrGolS:棉花Gossypium raimondii,XP_012490292.1;VvGolS:葡萄Vitis vinifera,XP_002281369.1;SlGolS:番茄Solanum lycopersicum,NP_001234668.2;StGolS:马铃薯Solanum tuberosum,ADW78838.1;SbGolS:高粱Sorghum bicolor,XP_002467954.1;BdGolS:二穗短柄草Brachypodium distachyon,XP_003558046.1;SitGolS:谷子Setaria italica,XP_004984579.1;OsGolS:水稻Oryza sativa,XP_015628833.1;ZmGolS:玉米Zea mays,NP_001105748.2。划线处为Glycosyl transferase, family 8(IPR002495)保守结构域The line marks the Glycosyl transferase, family 8 (IPR002495) conserved domain"
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