|Table of Contents|

Agrobacterium rhizogenes Mediated Transformation System of Hengshan Astragalus membranaceus and Determination of Active Components

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2022年09
Page:
93-99
Research Field:
Publishing date:

Info

Title:
Agrobacterium rhizogenes Mediated Transformation System of Hengshan Astragalus membranaceus and Determination of Active Components
Author(s):
ZHANG Hongchi; LIU Rui; GAO Zhihui; LI Hui; YANG Yang
(College of Life Science,Shanxi Datong University,Datong,Shanxi 037009)
Keywords:
Hengshan Astragalus membranaceus; hairy roots; Agrobacterium rhizogenes; total flavonoids; total saponins
PACS:
-
DOI:
10.11937/bfyy.20214067
Abstract:
Hengshan Astragalus membranaceu leaves were used as the experimental materials.The experimental method was used to induce hairy roots by Agrobacterium rhizogenes infection.The ability of different strains to induce hairy roots from leaves and the genetic transformation system of hairy roots were studied,including effect of different explants,bacterial concentration,co-cultivation time,antibiotic type and concentration,and acetosyringone concentration on hairy root induction rate and the contents of total flavonoids and total saponins in hairy roots cultured on large scale were determined,a stable hairy root system of Hengshan Astragalus membranaceus were established,in order to provide reference for establishing a liquid culture system of hairy roots that accumulate active ingredients.The results showed that the induction rate of hairy root was higher when the leaves were used as the explant,soaked for 30 minutes by OD600=0.6 LBA9402,co-cultured for 3 days,cultured in 1/2MS medium containing 100 μmol?L-1 acetosyringone,selected on screening medium with 500 mg?L-1 cefixime.The induction rate of hairy root was 54.1%.It was confirmed by PCR analysis that the rooting gene in the LBA9402 was integrated into the genome of Astragalus membranaceus.The content of total flavonoids and total saponins of hairy roots increased significantly after liquid culture expansion,which were 1.72 times and 2.31 times that of natural roots,respectively.

References:

[1]卞云云,管佳,毕志明,等.蒙古黄芪的化学成分研究[J].中国药学杂志,2006,41(16):1217-1221.[2]张弘弛,刘瑞,巴德方,等.恒山黄芪1H-NMR指纹图谱的研究[J].食品工业科技,2022,43(1):47-55.[3]ZHANG H C,LIU R,ZHOU F.Chaetoglobosins in endophytic Chaetomium sp.from Astragalus membranaceus[J].Chemistry of Natural Compounds,2021,57:401-403.[4]RON M,KAJALA K,PAULUZZI G.Hairy root transformation using Agrobacterium rhizogenes as a tool for exploring cell type-specific gene expression and function using tomato as a model[J].Plant Physiology,2014,166:455-469.[5]GAI Q Y,JIAO J,WANG X,et al.Elicitation of Isatis tinctoria L.hairy root cultures by salicylic acid and methyl jasmonate for the enhanced production of pharmacologically active alkaloids and flavonoids[J].Plant Cell,Tissue and Organ Culture,2019,137:77-86.[6]CHEN L,CAI Y P,LIU X J,et al.Soybean hairy roots produced in vitro by Agrobacterium rhizogenes mediated transformation[J].The Crop Journal,2018,6(2):162-171.[7]姚庆收,陈向明,丁斐,等.甘草毛状根遗传转化体系的建立及甘草酸和总黄酮含量的测定[J].中药材,2018,41(9):2035-2038.[8]姚庆收,陈向明,马玉娥,等.蒙古黄芪毛状根的高效诱导及毛状根生物量和总黄酮含量的比较分析[J].中药材,2017,40(8):1753-1757.[9]贾春秋,赵雪.苦参毛状根的诱导培养及氧化苦参碱含量的测定[J].人参研究,2014,26(3):39-42.[10]REIS A,BOUTET-MERCEY S,MASSOT S,et al.Isoflavone production in hairy root cultures and plantlets of Trifolium pratense[J].Biotechnology Letters,2019,41:427-442.[11]梅错,刘志鹏.发根农杆菌介导的箭筈豌豆毛状根遗传转化体系的建立[J].中国草地学报,2020,42(5):1-7.[12]BOISSON-DERNIER A,CHABAUD M,GARCIA F,et al.Agrobacterium rhizogenes-transformed roots of Medicago truncatula for the study of nitrogen-fixing and endomycorrhizal symbiotic associations[J].Molecular Plant-Microbe Interactions,2001,14(6):695-700.[13]王天佐,张文浩.发根农杆菌介导的花苜蓿毛状根转化体系的建立[J].草地学报,2020,28(1):268-272.[14]AGGARWAL P R,NAG P,CHOUDHARY P,et al.Genotype-independent Agrobacterium rhizogenes mediated root transformation of chickpea:A rapid and efficient method for reverse genetics studies[J].Plant Methods,2018(14):55.[15]STILLER J,MARTIRANI L,TUPPALE S,et al.High frequency transformation and regeneration of transgenic plants in the model legume Lotus japonicus[J].Journal of Experimental Botany,1997,48(7):1357-1365.[16]徐悦,曹英萍,王玉,等.发根农杆菌介导的菠菜毛状根遗传转化体系的建立[J].植物学报,2019,54(4):515-521.[17]刘亚令,耿雅萍,黄小苏,等.基于HPLC法测定不同产地黄芪总黄酮和异黄酮的含量[J].山西农业大学学报(自然科学版),2019,39(5):54-61.[18]周超,何轶,林瑞超,等.黄芪4种皂苷提取物的制备及其在黄芪总皂苷含量测定中的应用[J].中国实验方剂学杂志,2013,19(22):56-59.[19]CAI Z Z,KASTELL A,KNORR D,et al.Exudation：an expanding tecnique for fontinouus production and release of seconeday metadata form plat cel suspenseion and hairy root courses[J].Plant Cell Biotechnology,2012,31(3):461-477.[20]翁颖妮,朱婧,王琴波,等.黄花倒水莲毛状根诱导及其愈伤组织总皂苷的测定[J].中药材,2020,43(9):2099-2102.

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Last Update: 2022-06-28