Towards delivering novel maize genotypes with improvedresilience and productivity through the application ofpredictive breeding technologies to modulate strigolactone levels
Başlık çevirisi mevcut değil.
- Tez No: 914164
- Danışmanlar: DR. PAUL CHRİSTOU
- Tez Türü: Yüksek Lisans
- Konular: Belirtilmemiş.
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2022
- Dil: İngilizce
- Üniversite: Instituto Agronómico Mediterráneo de Zaragoza
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 80
Özet
Maize (Z.mays) is the most cultivated cereal in the world. It originates from teosinte. Teosinte has undergone a huge process of transformation and selection. Then, it became modern corn, which people now grow. It has been cultivated in more than 160 countries, especially in tropical, subtropical, and temperate regions. Yields have increased steadily throughout the world since the 1930s thanks to the contribution of hybrid seeds, nitrogen fertilizers, and herbicide. Over the years, maize has become an important food source for animals and humans. Recently, genome-editing technology has emerged as a powerful tool for tailoring targeted genes and creating precise mutations. Strigolactones are carotenoid-derived, secondary metabolites, and phytohormones. They are produced by the plant. Strigolactones have many roles in plants: they stimulate germination of parasitic weed seeds, modulate root architecture, inhibit shooting, improve resistance against abiotic stress conditions, and improve symbiotic Arbuscular Mycorrhiza Fungi (AMF), which helps the plant to improve root architecture to improve plant nutrition acquisition from the soil. In the biosynthesis of strigolactones, Carotenoid Cleavage Dioxygenase 7 (CCD7) and Carotenoid Cleavage Dioxygenase 8 (CCD8) are two genes responsible for strigolactone production. Upregulation of CCD7 and CCD8 expression can modulate shoot and root architecture in the plant, thereby improving nutrient uptake from the soil, especially under phosphate and nitrogen deficiency conditions. It is necessary to take into account that there is a strong relationship between phosphate, nitrogen and maize production. In our project, by using CRISPR/Cas9 engineering, we aimed to modulate the expression level of CCD7 and CCD8 genes in the promoter region. To do that, we targeted promoter regions of these two genes in the elite W37 inbred maize line to modulate strigolactone expression levels in maize by using two strategies: Staphylococcus aureus (SaCas9) and Staphylococcus pyogenes (SpCas9). We confirmed 153 lines positive for SaCas9 and 80 lines positive for SpCas9 for at least one gRNA combination at DNA level. However, we did not detect any mutations in maize with the codonoptimized SaCas9 and rice codon-optimized SpCas9, as well as the expected protein (45 kDa) accumulation, in our positive maize lines. We confirmed the transcription of SaCas9 and SpCas9. However, we did not confirm the translation of SaCas9 and SpCas9. Further experiments are needed in order to generate data to understand the necessary elements and strategy for improving the system.
Özet (Çeviri)
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