Development of multiplex genome editing toolkits for citrus with high efficacy in biallelic and homozygous mutations

Huang, Xiaoen; Wang, Yuanchun; Xu, Jin; Wang, Nian

doi:10.1007/s11103-020-01043-6

Cited by 60 publications

(69 citation statements)

References 40 publications

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“…Protoplast cells of C. sinensis ‘Hamlin’ were prepared as described by 13 . Embryogenic calli were subcultured on solid MT (Murashige and Tucker) media (Phytotech) every 2 weeks.…”

Section: Methodsmentioning

confidence: 99%

Citrus Huanglongbing is an immune-mediated disease that can be treated by mitigating reactive oxygen species triggered cell death of the phloem tissues caused by Candidatus Liberibacter asiaticus

Pang

Huang

et al. 2021

Preprint

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The immune system is critical for keeping animals and plants healthy from pathogens. However, immune-mediated diseases are also common for human. Immune-mediated diseases have not been reported for plants. Here, we present evidence that citrus Huanglongbing (HLB), caused by phloem-colonizing Candidatus Liberibacter asiaticus (CLas), is an immune-mediated disease. CLas infection of Citrus sinensis stimulated systemic and chronic immune response in the phloem tissues including reactive oxygen species (ROS) production as indicated by H2O2, callose deposition, and induction of immune related genes. Systemic cell death of companion and sieve element cells, but not surrounding parenchyma cells, was observed following ROS production triggered by CLas. ROS production triggered by CLas localized in phloem tissues. The H2O2 concentration in exudates extracted from phloem enriched bark tissue of CLas infected plants reached a threshold of killing citrus protoplast cells, which was suppressed by uric acid (a ROS scavenger) and gibberellin. Foliar spray of HLB positive citrus with antioxidants (uric acid and rutin) and gibberellin significantly reduced both H2O2 concentrations and cell death in phloem tissues induced by CLas and reduced HLB symptoms. RNA-seq analyses of CLas infected and health C. sinensis support that CLas causes oxidative stress. In sum, HLB is an immune-mediated disease and both mitigating ROS via antioxidants and promoting plant growth can reduce cell death of the phloem tissues caused by CLas, thus controlling HLB.

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“…Protoplast cells of C. sinensis ‘Hamlin’ were prepared as described by 13 . Embryogenic calli were subcultured on solid MT (Murashige and Tucker) media (Phytotech) every 2 weeks.…”

Section: Methodsmentioning

confidence: 99%

Citrus Huanglongbing is an immune-mediated disease that can be treated by mitigating reactive oxygen species triggered cell death of the phloem tissues caused by Candidatus Liberibacter asiaticus

Pang

Huang

et al. 2021

Preprint

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“…The ability to detect mutant phenotype at the early stages of plant growth for quickly checking the effectiveness of the selected conditions is especially valuable in experiments with perennial crops, which include fruit crops, most berry crops and grapes. This approach, based on the use of the marker gene PDS, has been successfully applied in the optimization of conditions for the CRISPR/Cas9 editing of plants as grapes, apple trees, bananas, strawberries, kiwi and citruses [22,23,32,41,43,44,[50][51][52][53][54][55][56][57][58][59][60][61] (Table S1).…”

Section: Optimization Of Editing With Marker Gene Usagementioning

confidence: 99%

“…Similar results were obtained by Huang et al on citrus (Citrus × sinensis (L.) Osbeck), showing a higher PTG/Cas9 constructs efficiency for PDS knockout. As a result, biallelic mutations with a 44.4% rate and homozygous mutations with a 11.1% rate were obtained, and the possibility of obtaining large deletions (15 kb) using CRISPR/Cas9 was demonstrated [32]. Zhu et al used a marker gene knockout to test editing conditions on another citrus crop kumquat (Fortunella margarita (Lour.)…”

Section: Optimization Of Editing With Marker Gene Usagementioning

confidence: 99%

“…Genome editing of first fruit and berry cultures has been conducted by CRISPR/Cas9 fairly recent [23][24][25]. This review summarizes the results of CRISPR/Cas9 genome editing studies of different groups of target genes in fruit and berry crops: (1) in order to decrease biotic and abiotic stress susceptibility [19,[24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39], (2) change flowering time and plant architectonics [40][41][42][43][44][45], (3) increase shelf-time [46], (4) study gene functions [43][44][45][47][48][49] and (5) optimize CRISPR/Cas9 editing of fruit and berry cultures by using the marker [22,23,32,41,43,44,[50][51][52][53][54][55][56][57]…”

Section: Introductionmentioning

confidence: 99%

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Applications of CRISPR/Cas9 System in Vegetatively Propagated Fruit and Berry Crops

et al. 2021

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Fruit and berry crops, as well as grapes, are important parts of the human diet and, at the same time, significant objects of genetic, breeding, biochemical and nutritional research. Traditional approaches of crop research and improvement are now complemented by effective modern genetic technologies. In this review, we analyze and summarize the achievements in genome editing of fruit, berry crops and grapes. New approaches accelerate the improvement of genotypes for many groups of traits: plant resistance to unfavorable environmental factors, flowering and ripening time, plant architectonics, fruit shelf time and biochemical composition. Genome editing using the CRISPR/Cas9 system has been successfully tested on the most important vegetatively propagated fruit and berry crops (apple, pear, orange, kumquat, grapefruit, banana, strawberry and kiwi) and grapes. About 30 genes of these crops have been used as targets for the introduction of desired mutations using the CRISPR/Cas9 system. The most valuable results are the improvement of important agronomic traits. For 24 genes it has been shown that their knockout can result in the improvement of varieties. In addition, the review pays attention to the comparative analysis of the explant types of vegetatively propagated crops used for the delivery of editing genetic constructs, as well as the comparison of the editing efficiency depending on the variation of the objects used, delivery methods, etc. The article discusses the existing limitations that need to be overcome for a wider application of genomic editing in order to improve varieties of fruit and berry crops, as well as grapes.

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“…1) (15). Fortunately, this technology has been successfully used to edit citrus genomes and generate disease-resistant citrus varieties (16)(17)(18)(19)(20).…”

mentioning

confidence: 99%

A promising plant defense peptide against citrus Huanglongbing disease

Wang

2021

Proc. Natl. Acad. Sci. U.S.A.

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Development of multiplex genome editing toolkits for citrus with high efficacy in biallelic and homozygous mutations

Cited by 60 publications

References 40 publications

Citrus Huanglongbing is an immune-mediated disease that can be treated by mitigating reactive oxygen species triggered cell death of the phloem tissues caused by Candidatus Liberibacter asiaticus

Citrus Huanglongbing is an immune-mediated disease that can be treated by mitigating reactive oxygen species triggered cell death of the phloem tissues caused by Candidatus Liberibacter asiaticus

Applications of CRISPR/Cas9 System in Vegetatively Propagated Fruit and Berry Crops

A promising plant defense peptide against citrus Huanglongbing disease

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