CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea
Cyclic nucleotide-gated ion channels (CNGCs) have been reported to be involved in multiple plant physiological processes. Their involvement in plant immunity has been studied in several herbal plant species. It remains unclear whether CNGCs in woody plants play a similar role in plant immunity. In t...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2020.575477/full |
| _version_ | 1818261370046513152 |
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| author | Huijuan Zhou Huijuan Zhou Huijuan Zhou Suhua Bai Suhua Bai Suhua Bai Nan Wang Nan Wang Nan Wang Xiaohong Sun Xiaohong Sun Yugang Zhang Yugang Zhang Jun Zhu Jun Zhu Chaohua Dong Chaohua Dong Chaohua Dong |
| author_facet | Huijuan Zhou Huijuan Zhou Huijuan Zhou Suhua Bai Suhua Bai Suhua Bai Nan Wang Nan Wang Nan Wang Xiaohong Sun Xiaohong Sun Yugang Zhang Yugang Zhang Jun Zhu Jun Zhu Chaohua Dong Chaohua Dong Chaohua Dong |
| author_sort | Huijuan Zhou |
| collection | DOAJ |
| description | Cyclic nucleotide-gated ion channels (CNGCs) have been reported to be involved in multiple plant physiological processes. Their involvement in plant immunity has been studied in several herbal plant species. It remains unclear whether CNGCs in woody plants play a similar role in plant immunity. In the present study, we identified an apple CNGC (designated as MdCNGC2), which is the homolog of Arabidopsis CNGC2. Analysis of tissue distribution revealed that MdCNGC2 was expressed in all tested tissues. Abundant transcripts of MdCNGC2 were observed in leaves and shoot bark. Low expression was observed in fruits and roots. MdCNGC2 expression was induced in apple callus and shoot bark by Botryosphaeria dothidea. The induction of MdCNGC2 was significantly higher in susceptible cultivars “Fuji,” “Ralls Janet,” and “Gala” compared to the resistant cultivar “Jiguan,” suggesting that MdCNGC2 may be a negative regulator of resistance to B. dothidea. MdCNGC2 mutagenesis mediated by gene editing based on the CRISPR/Cas9 system led to constitutive accumulation of SA in apple callus. A culture filtrate of B. dothidea (BCF) induced the expression of several defense-related genes including MdPR1, MdPR2, MdPR4, MdPR5, MdPR8, and MdPR10a. Moreover, the induction of these genes was significantly higher in mdcngc2 mutant (MUT) callus than in wild type (WT) callus. Further analysis showed that the spread of B. dothidea was significantly lower on MUT callus than on WT callus. Knockdown of the MdCNGC2 gene reduced lesions caused by B. dothidea in apple fruits. These results collectively indicate that MdCNGC2 is a negative regulator of resistance to B. dothidea in apple callus. |
| first_indexed | 2024-12-12T18:46:09Z |
| format | Article |
| id | doaj.art-4ef61a5d16404417abfb7fe42e282d5a |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-12T18:46:09Z |
| publishDate | 2020-11-01 |
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| series | Frontiers in Plant Science |
| spelling | doaj.art-4ef61a5d16404417abfb7fe42e282d5a2022-12-22T00:15:30ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2020-11-011110.3389/fpls.2020.575477575477CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothideaHuijuan Zhou0Huijuan Zhou1Huijuan Zhou2Suhua Bai3Suhua Bai4Suhua Bai5Nan Wang6Nan Wang7Nan Wang8Xiaohong Sun9Xiaohong Sun10Yugang Zhang11Yugang Zhang12Jun Zhu13Jun Zhu14Chaohua Dong15Chaohua Dong16Chaohua Dong17Key Laboratory of Plant Biotechnology of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaShandong Province Key Laboratory of Applied Mycology, Qingdao, ChinaKey Laboratory of Plant Biotechnology of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaShandong Province Key Laboratory of Applied Mycology, Qingdao, ChinaKey Laboratory of Plant Biotechnology of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaShandong Province Key Laboratory of Applied Mycology, Qingdao, ChinaKey Laboratory of Plant Biotechnology of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaCollege of Horticulture, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaCollege of Horticulture, Qingdao Agricultural University, Qingdao, ChinaKey Laboratory of Plant Biotechnology of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, ChinaQingdao Key Laboratory of Genetic Improvement and Breeding in Horticultural Plants, Qingdao Agricultural University, Qingdao, ChinaShandong Province Key Laboratory of Applied Mycology, Qingdao, ChinaCyclic nucleotide-gated ion channels (CNGCs) have been reported to be involved in multiple plant physiological processes. Their involvement in plant immunity has been studied in several herbal plant species. It remains unclear whether CNGCs in woody plants play a similar role in plant immunity. In the present study, we identified an apple CNGC (designated as MdCNGC2), which is the homolog of Arabidopsis CNGC2. Analysis of tissue distribution revealed that MdCNGC2 was expressed in all tested tissues. Abundant transcripts of MdCNGC2 were observed in leaves and shoot bark. Low expression was observed in fruits and roots. MdCNGC2 expression was induced in apple callus and shoot bark by Botryosphaeria dothidea. The induction of MdCNGC2 was significantly higher in susceptible cultivars “Fuji,” “Ralls Janet,” and “Gala” compared to the resistant cultivar “Jiguan,” suggesting that MdCNGC2 may be a negative regulator of resistance to B. dothidea. MdCNGC2 mutagenesis mediated by gene editing based on the CRISPR/Cas9 system led to constitutive accumulation of SA in apple callus. A culture filtrate of B. dothidea (BCF) induced the expression of several defense-related genes including MdPR1, MdPR2, MdPR4, MdPR5, MdPR8, and MdPR10a. Moreover, the induction of these genes was significantly higher in mdcngc2 mutant (MUT) callus than in wild type (WT) callus. Further analysis showed that the spread of B. dothidea was significantly lower on MUT callus than on WT callus. Knockdown of the MdCNGC2 gene reduced lesions caused by B. dothidea in apple fruits. These results collectively indicate that MdCNGC2 is a negative regulator of resistance to B. dothidea in apple callus.https://www.frontiersin.org/articles/10.3389/fpls.2020.575477/fullBotryosphaeria dothideagene editingimmune defenseMalus domesticaapple callus |
| spellingShingle | Huijuan Zhou Huijuan Zhou Huijuan Zhou Suhua Bai Suhua Bai Suhua Bai Nan Wang Nan Wang Nan Wang Xiaohong Sun Xiaohong Sun Yugang Zhang Yugang Zhang Jun Zhu Jun Zhu Chaohua Dong Chaohua Dong Chaohua Dong CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea Frontiers in Plant Science Botryosphaeria dothidea gene editing immune defense Malus domestica apple callus |
| title | CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea |
| title_full | CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea |
| title_fullStr | CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea |
| title_full_unstemmed | CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea |
| title_short | CRISPR/Cas9-Mediated Mutagenesis of MdCNGC2 in Apple Callus and VIGS-Mediated Silencing of MdCNGC2 in Fruits Improve Resistance to Botryosphaeria dothidea |
| title_sort | crispr cas9 mediated mutagenesis of mdcngc2 in apple callus and vigs mediated silencing of mdcngc2 in fruits improve resistance to botryosphaeria dothidea |
| topic | Botryosphaeria dothidea gene editing immune defense Malus domestica apple callus |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2020.575477/full |
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