CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato
Recently, CRISPR-Cas9-based genome editing has been widely used for plant breeding. In our previous report, a tomato gene encoding hybrid proline-rich protein 1 (HyPRP1), a negative regulator of salt stress responses, has been edited using a CRISPR-Cas9 multiplexing approach that resulted in precise...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2023.1186932/full |
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| author | Mil Thi Tran Mil Thi Tran Mil Thi Tran Geon Hui Son Young Jong Song Ngan Thi Nguyen Seonyeong Park Thanh Vu Thach Jihae Kim Yeon Woo Sung Swati Das Dibyajyoti Pramanik Jinsu Lee Ki-Ho Son Sang Hee Kim Sang Hee Kim Tien Van Vu Tien Van Vu Jae-Yean Kim Jae-Yean Kim Jae-Yean Kim |
| author_facet | Mil Thi Tran Mil Thi Tran Mil Thi Tran Geon Hui Son Young Jong Song Ngan Thi Nguyen Seonyeong Park Thanh Vu Thach Jihae Kim Yeon Woo Sung Swati Das Dibyajyoti Pramanik Jinsu Lee Ki-Ho Son Sang Hee Kim Sang Hee Kim Tien Van Vu Tien Van Vu Jae-Yean Kim Jae-Yean Kim Jae-Yean Kim |
| author_sort | Mil Thi Tran |
| collection | DOAJ |
| description | Recently, CRISPR-Cas9-based genome editing has been widely used for plant breeding. In our previous report, a tomato gene encoding hybrid proline-rich protein 1 (HyPRP1), a negative regulator of salt stress responses, has been edited using a CRISPR-Cas9 multiplexing approach that resulted in precise eliminations of its functional domains, proline-rich domain (PRD) and eight cysteine-motif (8CM). We subsequently demonstrated that eliminating the PRD domain of HyPRP1 in tomatoes conferred the highest level of salinity tolerance. In this study, we characterized the edited lines under several abiotic and biotic stresses to examine the possibility of multiple stress tolerance. Our data reveal that the 8CM removal variants of HK and the KO alleles of both HK and 15T01 cultivars exhibited moderate heat stress tolerance. Similarly, plants carrying either the domains of the PRD removal variant (PR1v1) or 8CM removal variants (PR2v2 and PR2v3) showed better germination under osmosis stress (up to 200 mM mannitol) compared to the WT control. Moreover, the PR1v1 line continuously grew after 5 days of water cutoff. When the edited lines were challenged with pathogenic bacteria of Pseudomonas syringae pv. tomato (Pto) DC3000, the growth of the bacterium was significantly reduced by 2.0- to 2.5-fold compared to that in WT plants. However, the edited alleles enhanced susceptibility against Fusarium oxysporum f. sp. lycopersici, which causes fusarium wilt. CRISPR-Cas9-based precise domain editing of the SlHyPRP1 gene generated multi-stress-tolerant alleles that could be used as genetic materials for tomato breeding. |
| first_indexed | 2024-04-09T12:38:38Z |
| format | Article |
| id | doaj.art-652459ccbd364b0fb91545c2abf25f3a |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-09T12:38:38Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-652459ccbd364b0fb91545c2abf25f3a2023-05-15T05:06:55ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2023-05-011410.3389/fpls.2023.11869321186932CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomatoMil Thi Tran0Mil Thi Tran1Mil Thi Tran2Geon Hui Son3Young Jong Song4Ngan Thi Nguyen5Seonyeong Park6Thanh Vu Thach7Jihae Kim8Yeon Woo Sung9Swati Das10Dibyajyoti Pramanik11Jinsu Lee12Ki-Ho Son13Sang Hee Kim14Sang Hee Kim15Tien Van Vu16Tien Van Vu17Jae-Yean Kim18Jae-Yean Kim19Jae-Yean Kim20Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaCrop Science and Rural Development Division, College of Agriculture, Bac Lieu University, Bac Lieu, VietnamDivision of Horticultural Science, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Horticultural Science, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Life Science, Gyeongsang National University, Jinju, Republic of KoreaDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaNational Key Laboratory for Plant Cell Biotechnology, Agricultural Genetics Institute, Hanoi, VietnamDivision of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of KoreaDivision of Life Science, Gyeongsang National University, Jinju, Republic of KoreaNulla Bio R&D Center, Nulla Bio Inc., Jinju, Republic of KoreaRecently, CRISPR-Cas9-based genome editing has been widely used for plant breeding. In our previous report, a tomato gene encoding hybrid proline-rich protein 1 (HyPRP1), a negative regulator of salt stress responses, has been edited using a CRISPR-Cas9 multiplexing approach that resulted in precise eliminations of its functional domains, proline-rich domain (PRD) and eight cysteine-motif (8CM). We subsequently demonstrated that eliminating the PRD domain of HyPRP1 in tomatoes conferred the highest level of salinity tolerance. In this study, we characterized the edited lines under several abiotic and biotic stresses to examine the possibility of multiple stress tolerance. Our data reveal that the 8CM removal variants of HK and the KO alleles of both HK and 15T01 cultivars exhibited moderate heat stress tolerance. Similarly, plants carrying either the domains of the PRD removal variant (PR1v1) or 8CM removal variants (PR2v2 and PR2v3) showed better germination under osmosis stress (up to 200 mM mannitol) compared to the WT control. Moreover, the PR1v1 line continuously grew after 5 days of water cutoff. When the edited lines were challenged with pathogenic bacteria of Pseudomonas syringae pv. tomato (Pto) DC3000, the growth of the bacterium was significantly reduced by 2.0- to 2.5-fold compared to that in WT plants. However, the edited alleles enhanced susceptibility against Fusarium oxysporum f. sp. lycopersici, which causes fusarium wilt. CRISPR-Cas9-based precise domain editing of the SlHyPRP1 gene generated multi-stress-tolerant alleles that could be used as genetic materials for tomato breeding.https://www.frontiersin.org/articles/10.3389/fpls.2023.1186932/fullCRISPR-Cas9HyPRP1abiotic stressbiotic stressheat stress tolerancemulti-stress tolerance |
| spellingShingle | Mil Thi Tran Mil Thi Tran Mil Thi Tran Geon Hui Son Young Jong Song Ngan Thi Nguyen Seonyeong Park Thanh Vu Thach Jihae Kim Yeon Woo Sung Swati Das Dibyajyoti Pramanik Jinsu Lee Ki-Ho Son Sang Hee Kim Sang Hee Kim Tien Van Vu Tien Van Vu Jae-Yean Kim Jae-Yean Kim Jae-Yean Kim CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato Frontiers in Plant Science CRISPR-Cas9 HyPRP1 abiotic stress biotic stress heat stress tolerance multi-stress tolerance |
| title | CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato |
| title_full | CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato |
| title_fullStr | CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato |
| title_full_unstemmed | CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato |
| title_short | CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato |
| title_sort | crispr cas9 based precise engineering of slhyprp1 protein towards multi stress tolerance in tomato |
| topic | CRISPR-Cas9 HyPRP1 abiotic stress biotic stress heat stress tolerance multi-stress tolerance |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2023.1186932/full |
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