植物研究 ›› 2023, Vol. 43 ›› Issue (5): 779-786.doi: 10.7525/j.issn.1673-5102.2023.05.014
霍清清, 夏雨新, 李佳乐, 韩薇, 张书雅, 张哲, 夏美玲, 郭雯华, 由香玲()
收稿日期:
2023-03-19
出版日期:
2023-09-20
发布日期:
2023-09-05
通讯作者:
由香玲
E-mail:youxiangling@nefu.edu.cn
作者简介:
霍清清(2002—),女,本科,主要从事药用植物细胞工程研究。
基金资助:
Qingqing HUO, Yuxin XIA, Jiale LI, Wei HAN, Shuya ZHANG, Zhe ZHANG, Meiling XIA, Wenhua GUO, Xiangling YOU()
Received:
2023-03-19
Online:
2023-09-20
Published:
2023-09-05
Contact:
Xiangling YOU
E-mail:youxiangling@nefu.edu.cn
About author:
E-mail:youxiangling@nefu.edu.cnSupported by:
摘要:
为探讨龙牙楤木(Aralia elata)三萜皂苷合成的关键酶基因β-香树素合成酶基因(Aeβ-AS)对三萜皂苷合成的影响。利用基因克隆技术对Aeβ-AS基因进行克隆,并对烟草进行遗传转化,分析转基因烟草中Aeβ-AS基因在各部位的表达差异,检测Aeβ-AS基因及上下游关键酶基因表达量和总三萜含量。结果表明:成功构建了植物过表达载体pROKⅡ-Aeβ-AS,并转入了野生型烟草,获得7个转基因株系,并在mRNA水平表达,且在叶子中的表达量要高于根和茎;在转基因烟草中,Aeβ-AS基因及其上下游的关键酶基因表达量均高于野生型,其中株系L21的NtFPS、NtSS基因的相对表达量最高,株系L30的NtSE、Aeβ-AS基因的相对表达量最高;与野生型烟草相比,转基因烟草的总三萜含量显著升高(1.1~1.6倍)。试验结果证明合成Aeβ-AS基因并在烟草中进行异源转化,可以使转基因烟草内的总三萜含量显著升高。
中图分类号:
霍清清, 夏雨新, 李佳乐, 韩薇, 张书雅, 张哲, 夏美玲, 郭雯华, 由香玲. 龙牙楤木Aeβ-AS基因的表达对烟草中皂苷含量的影响[J]. 植物研究, 2023, 43(5): 779-786.
Qingqing HUO, Yuxin XIA, Jiale LI, Wei HAN, Shuya ZHANG, Zhe ZHANG, Meiling XIA, Wenhua GUO, Xiangling YOU. Effect of Expression of Aeβ-AS gene from Aralia elata on the Content of Saponins in Nicotiana tabacum L.[J]. Bulletin of Botanical Research, 2023, 43(5): 779-786.
表1
试验中所用的引物
引物名称 Primer names | 引物序列5′→3′ Primer sequences 5′→3′ |
---|---|
Aeβ-AS-F | gagaaacacgggggactctagaATGTGGAAGCTTAAGATAGCGAAA |
Aeβ-AS-R | gggaaattcgagctcggtaccTTAGGTGCCTAGGGACGGTAAT |
pROKⅡ-F | AAGACCGGCAACAGGATTC |
pROKⅡ-R | CGCACAATCCCACTATCCTT |
RT-NtActin-F | CCTGAGGTCCTTTTCCAACCA |
RT-NtActin-R | GGATTCCGGCAGCTTCCATT |
RT-Aeβ-AS-F | CTTCCTATGCACCCAGCTAAA |
RT-Aeβ-AS-R | CCCAGAGCAGGTCTTGTATTT |
RT-NtFPS-R | TACTTCTCCGCAACCACATC |
RT-NtFPS-R | GGACAATCCGGCGATGAATA |
RT-NtSS-R | GTACCCAAGTGCCGTATGTT |
RT-NtSS-R | TTGCCATGACCTGTGGAATAG |
RT-NtSE-R | CAGATCCTTCGCCCATCTTATTT |
RT-NtSE-R | CAGCTGGCATGCTCCTATTT |
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