Scholarlink

<p><i>Camelina sativa</i> is an emerging low-resource oilseed crop that has significant potential for oil production. It is imperative to modify camelina oils for idealized fatty acid structure that can match diverse application needs. Camelina seed harbors considerable quantities of very long-chain fatty acids (VLCFAs) that may not be advantageous. These VLCFAs can be efficiently diminished by disabling the fatty acid elongase1 (FAE1) in camelina. The allohexaploid camelina possesses three alleles of FAE1 genes. Using CRISPR-Cas<sub>9</sub> technology, three alleles of FAE1 genes can be knocked out simultaneously. In this study, pFGC-pcoCas<sub>9</sub> recombinant expression vector containing gRNA designed for FAE1 gene silencing was used. In this research, firstly, somatic embryogenesis improvement and micropropagation of terminal shoot cultures were established from germinated seeds under in vitro conditions and used as target tissues for pFGC-pcoCas<sub>9</sub> recombinant expression vector containing gRNA designed for FAE1 silencing. The constructed recombinant vector was conveyed to the camelina plant by <i>Agrobacterium tumefaciens</i>. Three explants (hypocotyl, cotyledon, and terminal leaves) were utilized with five distinct combinations of plant growth regulators (NAA, BAP, 2,4-D, TDZ, and Kin) to improve somatic embryogenesis and regeneration in <i>Camelina</i>. The greatest rate of somatic embryogenesis was observed in the cotyledon explants cultivated in the MS medium including 0.5&#xa0;mg/L NAA + 5&#xa0;mg/L BAP, and the maximum frequency of regeneration was recorded in the terminal leaf explant cultured in the MS medium containing 0.5&#xa0;mg/L NAA + 2&#xa0;mg/L BAP + 1&#xa0;mg/L Kin. Transgenic <i>C. sativa</i> plants created by <i>Agrobacterium</i> transformation were confirmed by PCR technique. The genetic transformation efficiency reached 93.62%, indicating a high success rate in gene editing. This investigation illustrated that the CRISPR/Cas<sub>9</sub> system can be effectively applied to the polyploid crop camelina to quickly acquire features such as an ideal fatty acid profile in oil content. Gene knockout of FAE1 also provides a means to increase the levels of oleic acid or α-linolenic acid in camelina oils that are preferable for commercial or food/feed applications in the future.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Optimization of Somatic Embryogenesis and FAE1 Gene Silencing via CRISPR-Cas9 in Camelina sativa L.

  • 期刊论文

  • 发表日期：

    2025年8月21日

查看全文
  • Baghi Hamideh,
  • Tarinejad Alireza,
  • Pazhouhandeh Maghsoud,
  • Mahna Nasser

摘要

Camelina sativa is an emerging low-resource oilseed crop that has significant potential for oil production. It is imperative to modify camelina oils for idealized fatty acid structure that can match diverse application needs. Camelina seed harbors considerable quantities of very long-chain fatty acids (VLCFAs) that may not be advantageous. These VLCFAs can be efficiently diminished by disabling the fatty acid elongase1 (FAE1) in camelina. The allohexaploid camelina possesses three alleles of FAE1 genes. Using CRISPR-Cas9 technology, three alleles of FAE1 genes can be knocked out simultaneously. In this study, pFGC-pcoCas9 recombinant expression vector containing gRNA designed for FAE1 gene silencing was used. In this research, firstly, somatic embryogenesis improvement and micropropagation of terminal shoot cultures were established from germinated seeds under in vitro conditions and used as target tissues for pFGC-pcoCas9 recombinant expression vector containing gRNA designed for FAE1 silencing. The constructed recombinant vector was conveyed to the camelina plant by Agrobacterium tumefaciens. Three explants (hypocotyl, cotyledon, and terminal leaves) were utilized with five distinct combinations of plant growth regulators (NAA, BAP, 2,4-D, TDZ, and Kin) to improve somatic embryogenesis and regeneration in Camelina. The greatest rate of somatic embryogenesis was observed in the cotyledon explants cultivated in the MS medium including 0.5 mg/L NAA + 5 mg/L BAP, and the maximum frequency of regeneration was recorded in the terminal leaf explant cultured in the MS medium containing 0.5 mg/L NAA + 2 mg/L BAP + 1 mg/L Kin. Transgenic C. sativa plants created by Agrobacterium transformation were confirmed by PCR technique. The genetic transformation efficiency reached 93.62%, indicating a high success rate in gene editing. This investigation illustrated that the CRISPR/Cas9 system can be effectively applied to the polyploid crop camelina to quickly acquire features such as an ideal fatty acid profile in oil content. Gene knockout of FAE1 also provides a means to increase the levels of oleic acid or α-linolenic acid in camelina oils that are preferable for commercial or food/feed applications in the future.

Graphical Abstract