Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i>
Huanglongbing (HLB), also known as citrus greening, is the most destructive disease of citrus worldwide. In the United States, this disease is associated with a phloem-restricted bacterium, <i>Candidatus</i> Liberibacter asiaticus. Commercial citrus cultivars are susceptible to HLB, but...
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MDPI AG
2020-06-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/7/821 |
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| author | Xiaobao Ying Bryce Redfern Frederick G. Gmitter Zhanao Deng |
| author_facet | Xiaobao Ying Bryce Redfern Frederick G. Gmitter Zhanao Deng |
| author_sort | Xiaobao Ying |
| collection | DOAJ |
| description | Huanglongbing (HLB), also known as citrus greening, is the most destructive disease of citrus worldwide. In the United States, this disease is associated with a phloem-restricted bacterium, <i>Candidatus</i> Liberibacter asiaticus. Commercial citrus cultivars are susceptible to HLB, but <i>Poncirus trifoliata</i>, a close relative of <i>Citrus</i>, is highly tolerant of HLB. Isolating <i>P. trifoliata</i> gene(s) controlling its HLB tolerance followed by expressing the gene(s) in citrus is considered a potential cisgenic approach to engineering citrus for tolerance to HLB. Previous gene expression studies indicated that the <i>constitutive disease resistance</i> (<i>CDR</i>) genes in <i>P. trifoliata</i> (<i>PtCDRs</i>) may play a vital role in its HLB tolerance. This study was designed to use <i>Arabidopsis</i> mutants as a model system to confirm the function of <i>PtCDRs</i> in plant disease resistance. <i>PtCDR2</i> and <i>PtCDR8</i> were amplified from <i>P. trifoliata</i> cDNA and transferred into the <i>Arabidopsis cdr1</i> mutant, whose resident <i>CDR1</i> gene was disrupted by T-DNA insertion. The <i>PtCDR2</i> and <i>PtCDR8</i> transgenic <i>Arabidopsis cdr1</i> mutant restored its hypersensitive response to the bacterial pathogen <i>Pseudomonas syringae</i> pv. <i>tomato</i> strain DC3000 (<i>Pst</i> DC3000) expressing <i>avrRpt2</i>. The defense marker gene <i>PATHOGENESIS RELATED 1</i> (<i>PR1</i>) expressed at much higher levels in the <i>PtCDR2</i> or <i>PtCDR8</i> transgenic <i>cdr1</i> mutant than in the non-transgenic <i>cdr1</i> mutant with or without pathogen infection. Multiplication of <i>Pst</i> DC3000 bacteria in <i>Arabidopsis</i> was inhibited by the expression of <i>PtCDR2</i> and <i>PtCDR8</i>. Our results showed that <i>PtCDR2</i> and <i>PtCDR8</i> were functional in <i>Arabidopsis</i> and played a positive role in disease resistance and demonstrated that <i>Arabidopsis</i> mutants can be a useful alternate system for screening <i>Poncirus</i> genes before making the time-consuming effort to transfer them into citrus, a perennial woody plant that is highly recalcitrant for <i>Agrobacterium</i> or biolistic-mediated transformation. |
| first_indexed | 2024-03-10T18:48:08Z |
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| last_indexed | 2024-03-10T18:48:08Z |
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| spelling | doaj.art-224652bfb8f24a19ab0a30c2f2b0a4c72023-11-20T05:22:13ZengMDPI AGPlants2223-77472020-06-019782110.3390/plants9070821Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i>Xiaobao Ying0Bryce Redfern1Frederick G. Gmitter2Zhanao Deng3Gulf Coast Research and Education Center, IFAS, University of Florida, 14625 County Road 672, Wimauma, FL 33598, USAGulf Coast Research and Education Center, IFAS, University of Florida, 14625 County Road 672, Wimauma, FL 33598, USACitrus Research and Education Center, IFAS, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USAGulf Coast Research and Education Center, IFAS, University of Florida, 14625 County Road 672, Wimauma, FL 33598, USAHuanglongbing (HLB), also known as citrus greening, is the most destructive disease of citrus worldwide. In the United States, this disease is associated with a phloem-restricted bacterium, <i>Candidatus</i> Liberibacter asiaticus. Commercial citrus cultivars are susceptible to HLB, but <i>Poncirus trifoliata</i>, a close relative of <i>Citrus</i>, is highly tolerant of HLB. Isolating <i>P. trifoliata</i> gene(s) controlling its HLB tolerance followed by expressing the gene(s) in citrus is considered a potential cisgenic approach to engineering citrus for tolerance to HLB. Previous gene expression studies indicated that the <i>constitutive disease resistance</i> (<i>CDR</i>) genes in <i>P. trifoliata</i> (<i>PtCDRs</i>) may play a vital role in its HLB tolerance. This study was designed to use <i>Arabidopsis</i> mutants as a model system to confirm the function of <i>PtCDRs</i> in plant disease resistance. <i>PtCDR2</i> and <i>PtCDR8</i> were amplified from <i>P. trifoliata</i> cDNA and transferred into the <i>Arabidopsis cdr1</i> mutant, whose resident <i>CDR1</i> gene was disrupted by T-DNA insertion. The <i>PtCDR2</i> and <i>PtCDR8</i> transgenic <i>Arabidopsis cdr1</i> mutant restored its hypersensitive response to the bacterial pathogen <i>Pseudomonas syringae</i> pv. <i>tomato</i> strain DC3000 (<i>Pst</i> DC3000) expressing <i>avrRpt2</i>. The defense marker gene <i>PATHOGENESIS RELATED 1</i> (<i>PR1</i>) expressed at much higher levels in the <i>PtCDR2</i> or <i>PtCDR8</i> transgenic <i>cdr1</i> mutant than in the non-transgenic <i>cdr1</i> mutant with or without pathogen infection. Multiplication of <i>Pst</i> DC3000 bacteria in <i>Arabidopsis</i> was inhibited by the expression of <i>PtCDR2</i> and <i>PtCDR8</i>. Our results showed that <i>PtCDR2</i> and <i>PtCDR8</i> were functional in <i>Arabidopsis</i> and played a positive role in disease resistance and demonstrated that <i>Arabidopsis</i> mutants can be a useful alternate system for screening <i>Poncirus</i> genes before making the time-consuming effort to transfer them into citrus, a perennial woody plant that is highly recalcitrant for <i>Agrobacterium</i> or biolistic-mediated transformation.https://www.mdpi.com/2223-7747/9/7/821Huanglongbingdisease resistancecisgenicsgenetic engineering<i>Poncirus</i><i>Arabidopsis</i> |
| spellingShingle | Xiaobao Ying Bryce Redfern Frederick G. Gmitter Zhanao Deng Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> Plants Huanglongbing disease resistance cisgenics genetic engineering <i>Poncirus</i> <i>Arabidopsis</i> |
| title | Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> |
| title_full | Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> |
| title_fullStr | Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> |
| title_full_unstemmed | Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> |
| title_short | Heterologous Expression of the <i>Constitutive Disease Resistance 2</i> and <i>8</i> Genes from <i>Poncirus trifoliata</i> Restored the Hypersensitive Response and Resistance of <i>Arabidopsis cdr1</i> Mutant to Bacterial Pathogen <i>Pseudomonas syringae</i> |
| title_sort | heterologous expression of the i constitutive disease resistance 2 i and i 8 i genes from i poncirus trifoliata i restored the hypersensitive response and resistance of i arabidopsis cdr1 i mutant to bacterial pathogen i pseudomonas syringae i |
| topic | Huanglongbing disease resistance cisgenics genetic engineering <i>Poncirus</i> <i>Arabidopsis</i> |
| url | https://www.mdpi.com/2223-7747/9/7/821 |
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