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DAD1 gene editing in Arabidopsis and rapeseed towards inducing sterility
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Rahil Dolatabadi    , Abdolreza Bagheri * , Seyyed Hasan Marashi , Saied Malekzadeh Shafaroudi |
| Biotechnology and Plant Breeding Department, Ferdowsi University of Mashhad, Iran , bagheriyazd@gmail.com |
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Abstract: (447 Views) |
Hybrid seed production, which is one of the effective strategies to increase crop yield, relies on production of male sterile lines. The objective of this study was to investigate the possibility of producing male sterile lines in rapeseed using CRISPR technology. Different genes involved in the development of pollen and anthers were analyzed and the DAD1 gene, which encodes a chloroplast phospholipase and has two copies in the rapeseed genome, was selected as the target gene. After designing the CRISPR construct for this gene, the efficacy of this construct was investigated in Arabidopsis and rapeseed plants. In total, 18 Arabidopsis plants and 8 transgenic rapeseed plants containing the CRISPR construct were obtained, but none of the putative transgenic rapeseed plants reached the stage of seed production, and the putative transgenic Arabidopsis plants produced empty capsules. To check the effect of the CRISPR construct, DNA from three putative transgenic rapeseed plants was extracted and the sequence in the desired region of the DAD1 gene was analyzed. Sequencing results showed that deletions and additions have occurred in the target site of the CRISPR construct in the genomes of these 3 lines, and the lack of seed production in transgenic plants may be due to the failure of the DAD1 gene. The results of this study indicate that it is possible to achieve sterility using site-directed mutagenesis in DAD1 gene. To confirm the results and restore fertility to these lines, more investigations using inducible promoters for driving the CRISPR construct are required. |
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| Keywords: Anther, CRISPR, Hybrid seed |
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Full-Text [PDF 708 kb]
(261 Downloads)
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Type of Study: Research |
Subject:
Plant
Received: 2024/01/21 | Accepted: 2024/04/29 | Published: 2024/08/28
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Dolatabadi R, Bagheri A, Marashi S H, Malekzadeh Shafaroudi S. DAD1 gene editing in Arabidopsis and rapeseed towards inducing sterility. gebsj 2024; 13 (1) :1-9 URL: http://gebsj.ir/article-1-482-en.html
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