Induction and optimization of hairy root growth condition for Silybum marianum L. through inoculation by Agrobacterium rhizogenes inoculation
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Mohammad Ali Aazami *, Elham Rahmani, Mohammad Bagher Hassanpouraghdam |
Department of Horticulture, Faculty of Agriculture, Maragheh University, Iran , aazami58@gmail.com |
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Abstract: (214 Views) |
Silybum marianum L. from the Asteraceae is an annual or biennial plant widely distributed at the North, west and south of Iran. The fruits are containing medicinal flavonoids such as silymarin. This study was conducted to optimize the growing conditions for hairy roots and to assay the diverse factors affecting the induction and growth of roots induced by Agrobacterium rhizogenes. The seeds were cultured on MS medium and were kept in growth chamber. After germination and seedling growth, explants of leaves, hypocotyl and roots were taken after 5, 10 and 15 days. The explants were inoculated (treated) with two (R1000 and 15834) Agrobacterium rhizogenes strains. Furthermore, the effects of two inoculation methods (floating and injection) were assayed on the hairy roots emergence percent. MS and 1/2MS growing media were compared regarding the hairy roots establishment in liquid medium. In a separate experiment, the effect of sucrose amount (20, 30 and 40 gl-1) were evaluated on hairy root induction. The roots emergence percent was recorded. To verify the transgenic identity, PCR test with specific marker genes was carried out. The results revealed that, 15834 strain was the strain of choice in 5 days old hypocotyl explants and produced more hairy roots. The best rooting percentage was belonged to leaf explants during 15 days treated with R1000 strained Agrobacterium. Floating was the method of choice for inoculation. 30 gL-1 sucrose was the best treatment for the root induction as well. |
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Keywords: Agrobacterium rhizogenes, Explant, Hairy root, Silybum marianum L., Secondary Metabolite |
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Type of Study: Research |
Subject:
Divers Received: 2019/12/15 | Accepted: 2020/02/14 | Published: 2020/08/10
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