Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis
Somatic embryogenesis is valuable for clonal propagation and genetic improvement, and it also serves as an ideal system for studying plant development mechanisms. In <i>Larix kaempferi</i>, microRNA171 and its target gene <i>L. kaempferi SCARECROW-LIKE6</i> (<i>LaSCL6&l...
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2024-10-01
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| author | Qiao-Lu Zang Zha-Long Ye Li-Wang Qi Wan-Feng Li |
| author_facet | Qiao-Lu Zang Zha-Long Ye Li-Wang Qi Wan-Feng Li |
| author_sort | Qiao-Lu Zang |
| collection | DOAJ |
| description | Somatic embryogenesis is valuable for clonal propagation and genetic improvement, and it also serves as an ideal system for studying plant development mechanisms. In <i>Larix kaempferi</i>, microRNA171 and its target gene <i>L. kaempferi SCARECROW-LIKE6</i> (<i>LaSCL6</i>), which has two alternative splicing variants, can regulate somatic embryogenesis; however, the underlying molecular mechanism is still unknown. In this study, we overexpressed these two <i>LaSCL6</i> variants in <i>Oryza sativa</i> and <i>Arabidopsis thaliana</i> and then used the RNA-Seq method to screen genes from <i>O. sativa</i> and <i>A. thaliana</i>, whose expression patterns are related to those of <i>LaSCL6</i> variants. The screened genes were then used to search <i>L. kaempferi</i> proteins to identify the candidate target genes of <i>LaSCL6</i>. After yeast one-hybrid and dual- luciferase transcriptional activity assays, <i>cytochrome P450</i>, <i>family 89</i>, <i>subfamily A</i>, <i>polypeptide 5</i> (<i>CYP89A5</i>), and <i>wall-associated receptor kinase-like 20</i> (<i>WAKL20</i>) were confirmed to be the target genes of <i>LaSCL6-var1</i>; in addition, <i>WAKL20</i> and <i>UDP-glycosyltransferase 85A3</i> (<i>UGT85A3</i>) were confirmed to be the target genes of LaSCL6-var2. Moreover, APETALA2-like protein 2, a transcription factor from the AP2/ERF family, was shown to interact with LaSCL6-var1 and LaSCL6-var2. Taken together, our results suggest a regulatory network of miR171-<i>LaSCL6</i>. The findings presented here not only provide novel insights into the regulation of the miR171-<i>LaSCL6</i> module but also explain the mechanism underlying larch somatic embryogenesis and other biological processes. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2024-10-01 |
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| spelling | doaj-art-cadd960e0acf4f088b59fe769d5ae0ec2024-11-08T14:39:32ZengMDPI AGPlants2223-77472024-10-011321307210.3390/plants13213072Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic EmbryogenesisQiao-Lu Zang0Zha-Long Ye1Li-Wang Qi2Wan-Feng Li3State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of the National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaSomatic embryogenesis is valuable for clonal propagation and genetic improvement, and it also serves as an ideal system for studying plant development mechanisms. In <i>Larix kaempferi</i>, microRNA171 and its target gene <i>L. kaempferi SCARECROW-LIKE6</i> (<i>LaSCL6</i>), which has two alternative splicing variants, can regulate somatic embryogenesis; however, the underlying molecular mechanism is still unknown. In this study, we overexpressed these two <i>LaSCL6</i> variants in <i>Oryza sativa</i> and <i>Arabidopsis thaliana</i> and then used the RNA-Seq method to screen genes from <i>O. sativa</i> and <i>A. thaliana</i>, whose expression patterns are related to those of <i>LaSCL6</i> variants. The screened genes were then used to search <i>L. kaempferi</i> proteins to identify the candidate target genes of <i>LaSCL6</i>. After yeast one-hybrid and dual- luciferase transcriptional activity assays, <i>cytochrome P450</i>, <i>family 89</i>, <i>subfamily A</i>, <i>polypeptide 5</i> (<i>CYP89A5</i>), and <i>wall-associated receptor kinase-like 20</i> (<i>WAKL20</i>) were confirmed to be the target genes of <i>LaSCL6-var1</i>; in addition, <i>WAKL20</i> and <i>UDP-glycosyltransferase 85A3</i> (<i>UGT85A3</i>) were confirmed to be the target genes of LaSCL6-var2. Moreover, APETALA2-like protein 2, a transcription factor from the AP2/ERF family, was shown to interact with LaSCL6-var1 and LaSCL6-var2. Taken together, our results suggest a regulatory network of miR171-<i>LaSCL6</i>. The findings presented here not only provide novel insights into the regulation of the miR171-<i>LaSCL6</i> module but also explain the mechanism underlying larch somatic embryogenesis and other biological processes.https://www.mdpi.com/2223-7747/13/21/3072Japanese larchmiR171<i>LaSCL6</i>somatic embryogenesisregulatory network |
| spellingShingle | Qiao-Lu Zang Zha-Long Ye Li-Wang Qi Wan-Feng Li Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis Plants Japanese larch miR171 <i>LaSCL6</i> somatic embryogenesis regulatory network |
| title | Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis |
| title_full | Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis |
| title_fullStr | Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis |
| title_full_unstemmed | Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis |
| title_short | Identification of Target Gene and Interacting Protein of Two <i>LaSCL6</i> Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis |
| title_sort | identification of target gene and interacting protein of two i lascl6 i alternative splicing variants provides novel insights into larch somatic embryogenesis |
| topic | Japanese larch miR171 <i>LaSCL6</i> somatic embryogenesis regulatory network |
| url | https://www.mdpi.com/2223-7747/13/21/3072 |
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