The Genetic Transformation of Sweet Orange (Citrus sinensis L. Osbeck) for Enhanced Resistance to Citrus Canker

Author(s):  
Lorena Noelia Sendin ◽  
María Paula Filippone
2017 ◽  
Vol 93 (6) ◽  
pp. 607-621 ◽  
Author(s):  
Lorena Noelia Sendín ◽  
Ingrid Georgina Orce ◽  
Rocío Liliana Gómez ◽  
Ramón Enrique ◽  
Carlos Froilán Grellet Bournonville ◽  
...  

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 972D-973
Author(s):  
Ahmad A. Omar ◽  
Wen-Yuan Song ◽  
James H. Graham ◽  
Jude W. Grosser

Citrus canker disease caused by the bacterial pathogen Xanthomonas axonopodis pv. citri is becoming a worldwide problem. Xa21 gene is a member of the Xa21 gene family of rice, which provides broad spectrum Xanthomonas resistance in rice. `Hamlin' sweet orange [Citrus sinensis (L.) Osbeck) is one of the leading commercial cultivars in Florida because of its high yield potential and early maturity. `Hamlin' also has a high regeneration capacity from protoplasts and is often used in transformation experiments. Since the citrus canker pathogen is in the same genus, this gene may have potential to function against canker in citrus. The wild-type Xa21 gene contains an intron, and there are some questions whether dicot plants can process genes containing monocot introns (the cDNA is intron-free). Plasmids DNA, encoding the non-destructive selectable marker EGFP (Enhanced Green Fluorescent Protein) gene and the cDNA of the Xa21 gene were transformed or co-transformed into `Hamlin' orange protoplasts using polyethylene glycol. More than 200 transgenic embryoids were recovered. More than 400 transgenic plants were developed from 75 independent transgenic events. PCR analysis revealed the presence of the cDNA of the Xa21 and the GFP genes in the transgenic plants. Some of the plants have the GFP only. Southern analysis is showing integration of the cDNA into different sites ranges from one to five sites. Western analysis is showing the expression of the cDNA of the Xa21 gene in the transgenic citrus plants. This is the first time that a gene from rice has been stably integrated and expressed in citrus plants. Canker challenge assay is in progress.


2010 ◽  
Vol 75 (1-2) ◽  
pp. 11-23 ◽  
Author(s):  
Li Yang ◽  
Chunhua Hu ◽  
Na Li ◽  
Jiayin Zhang ◽  
Jiawen Yan ◽  
...  

Plant Disease ◽  
2016 ◽  
Vol 100 (10) ◽  
pp. 1994-2000 ◽  
Author(s):  
Aline M. O. Gonçalves-Zuliani ◽  
Danielle S. Y. Nanami ◽  
Bruna R. Barbieri ◽  
Terezinha A. Guedes ◽  
Carlos A. Zanutto ◽  
...  

Asiatic citrus canker (ACC, caused by the bacterium Xanthomonas citri subsp. citri) is a destructive disease of citrus in Brazil and in several other citrus-producing countries. ACC management is problematic, and bactericides such as copper can be reasonably efficacious but do not completely control the disease. Furthermore, injury by citrus leaf miner (CLM) can exacerbate severity of ACC. Host resistance is the most desirable solution for management of ACC; however, evaluations of germplasm indicate that resistance is limited in many popular species and cultivars that are grown commercially. Limited evaluations have been made of sweet orange (Citrus sinensis) selections. We evaluated resistance of 25 Pêra sweet orange selections to X. citri subsp. citri by wound inoculation and measuring lesion diameter under greenhouse conditions (wound inoculation indicates mesophyll resistance which will be valuable in areas where CLM exists). ACC severity was assessed on the same 25 selections at three locations in the field in Brazil, relying on natural inoculum and conditions to cause disease. In the greenhouse experiments, the selections EEL, Bianchi/CC, Ipiguá, Olimpia, IAC 2000/1, and Ovale Siracusa consistently had the smallest diameter lesions, indicating greatest resistance, although differences in lesion diameter were small. Results from the field experiments were less conclusive, although EEL and Ovale Siracusa were consistently numerically least affected by ACC. These results indicate selections of sweet orange that might be preferable to consider in canker-prone areas in Brazil and elsewhere.


2009 ◽  
Vol 122 (1) ◽  
pp. 109-115 ◽  
Author(s):  
Janaynna Magalhães Barbosa-Mendes ◽  
Francisco de Assis Alves Mourão Filho ◽  
Armando Bergamin Filho ◽  
Ricardo Harakava ◽  
Steven V. Beer ◽  
...  

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Qin Long ◽  
Meixia Du ◽  
Junhong Long ◽  
Yu Xie ◽  
Jingyun Zhang ◽  
...  

AbstractPathological hypertrophy (cell enlargement) plays an important role in the development of citrus canker, but its regulators are largely unknown. Although WRKY22 is known to be involved in pathogen-triggered immunity and positively regulates resistance to bacterial pathogens in Arabidopsis, rice and pepper, the CRISPR/Cas9-mediated partial knockout of CsWRKY22 improves resistance to Xanthomonas citri subsp. citri (Xcc) in Wanjincheng orange (Citrus sinensis Osbeck). Here, we demonstrate that CsWRKY22 is a nucleus-localized transcriptional activator. CsWRKY22-overexpressing plants exhibited dwarf phenotypes that had wrinkled and thickened leaves and were more sensitive to Xcc, whereas CsWRKY22-silenced plants showed no visible phenotype changes and were more resistant to Xcc. Microscopic observations revealed that the overexpression of CsWRKY22 increased cell size in the spongy mesophyll. Transcriptome analysis showed that cell growth-related pathways, such as the auxin and brassinosteroid hormonal signaling and cell wall organization and biogenesis pathways, were significantly upregulated upon CsWRKY22 overexpression. Interestingly, CsWRKY22 activated the expression of CsLOB1, which is a key gene regulating susceptibility to citrus canker. We further confirmed that CsWRKY22 bound directly to the W-boxes just upstream of the transcription start site of CsLOB1 in vivo and in vitro. We conclude that CsWRKY22 enhances susceptibility to citrus canker by promoting host hypertrophy and CsLOB1 expression. Thus, our study provides new insights into the mechanism regulating pathological hypertrophy and the function of WRKY22 in citrus.


2017 ◽  
Vol 149 (4) ◽  
pp. 865-873 ◽  
Author(s):  
Adilson K. Kobayashi ◽  
Luiz Gonzaga E. Vieira ◽  
João Carlos Bespalhok Filho ◽  
Rui Pereira Leite ◽  
Luiz Filipe P. Pereira ◽  
...  

2002 ◽  
Vol 59 (1) ◽  
pp. 35-40 ◽  
Author(s):  
Weliton Antonio Bastos de Almeida ◽  
Francisco de Assis Alves Mourão Filho ◽  
Beatriz Madalena Januzzi Mendes ◽  
Adriana Pinheiro Martinelli Rodriguez

Exogenous genes can be introduced in plants by genetic transformation techniques. However, an efficient tissue culture system with high rates of plant recovery is necessary for gene introduction. This work aimed to define organogenesis and plant regeneration protocols for sweet orange varieties Natal, Valencia and Hamlin (Citrus sinensis L. Osbeck) and Rangpur lime (Citrus limonia L. Osbeck) which can be used in plant transformation experiments. Seeds of which teguments were removed, were germinated in vitro and maintained in the dark for three weeks, followed by one week at 16-h photoperiod (40 µmol m-2 s-1) and 27 ± 2°C. Organogenesis induction was done by introducing epicotyl segments in MT medium with 25 g L-1 sucrose and different BAP concentrations. After adventitious bud growth, the shoots were transferred to MT medium with either NAA or IBA (1 mg L-1), or absence of auxin, for rooting. The best results were obtained with 1 mg L-1 BAP for bud induction and 1 mg L-1 IBA for rooting for all three sweet orange cultivars. The use of 0.5-2.5 mg L-1 BAP, followed by 1 mg L-1 IBA were the best growth regulator combinations for bud induction and rooting, respectively, for 'Rangpur' lime. The protocols presented in this work are suitable for associations with genetic transformation experiments for these cultivars.


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