Efficient knockout of phytoene desaturase gene using CRISPR/Cas9 in melon

Hooghvorst, Isidre; López-Cristoffanini, Camilo; Nogués, Salvador

doi:10.1038/s41598-019-53710-4

Cited by 71 publications

(40 citation statements)

References 36 publications

(39 reference statements)

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“…(yam; Syombua et al, 2021 ), watermelon ( Tian et al, 2017 ), Musa spp. (banana/plantain; Kaur et al, 2018 ), Actinidia chinensis (kiwifruit; Wang et al, 2018 ), Cucumis melo (melon; Hooghvorst et al, 2019 ), Fortunella hindsii (Hongkong kumquat; Zhu et al, 2019 ), and sorghum ( Liu et al, 2019a ). Additionally, PDS knockout has been achieved in the first application of CRISPR in Setaria italica (foxtail millet) protoplasts ( Lin et al, 2018 ).…”

Section: Applications Of Gene Editing In Orphan Cropsmentioning

confidence: 99%

Current Advancements and Limitations of Gene Editing in Orphan Crops

Venezia

Krainer

2021

Front. Plant Sci.

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Gene editing provides precise, heritable genome mutagenesis without permanent transgenesis, and has been widely demonstrated and applied in planta. In the past decade, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) has revolutionized the application of gene editing in crops, with mechanistic advances expanding its potential, including prime editing and base editing. To date, CRISPR/Cas has been utilized in over a dozen orphan crops with diverse genetic backgrounds, leading to novel alleles and beneficial phenotypes for breeders, growers, and consumers. In conjunction with the adoption of science-based regulatory practices, there is potential for CRISPR/Cas-mediated gene editing in orphan crop improvement programs to solve a plethora of agricultural problems, especially impacting developing countries. Genome sequencing has progressed, becoming more affordable and applicable to orphan crops. Open-access resources allow for target gene identification and guide RNA (gRNA) design and evaluation, with modular cloning systems and enzyme screening methods providing experimental feasibility. While the genomic and mechanistic limitations are being overcome, crop transformation and regeneration continue to be the bottleneck for gene editing applications. International collaboration between all stakeholders involved in crop improvement is vital to provide equitable access and bridge the scientific gap between the world’s most economically important crops and the most under-researched crops. This review describes the mechanisms and workflow of CRISPR/Cas in planta and addresses the challenges, current applications, and future prospects in orphan crops.

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Section: Applications Of Gene Editing In Orphan Cropsmentioning

confidence: 99%

Current Advancements and Limitations of Gene Editing in Orphan Crops

Venezia

Krainer

2021

Front. Plant Sci.

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“…In cucurbits, there are no reports describing the application of ZNF nor TALENs, as far as the authors know. Nevertheless, CRISPR/Cas9 has been applied successfully in cucumber [27,28], watermelon [29][30][31] and melon [32].…”

Section: Genome Editing In Cucurbit Speciesmentioning

confidence: 99%

“…Several CRISPR/Cas9 binary vectors have been reported successful in cucumber, watermelon and melon, and may be used for other cucurbits (Table 1). However, Agrobacterium-transformation protocol is still the main bottleneck to apply genome editing in cucurbits [31,32].…”

Section: Genome Editing In Cucurbit Speciesmentioning

confidence: 99%

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Opportunities and Challenges in Doubled Haploids and Haploid Inducer-Mediated Genome-Editing Systems in Cucurbits

Hooghvorst

Nogués

2020

Agronomy

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Doubled haploids have played a major role in cucurbit breeding for the past four decades. In situ parthenogenesis via irradiated pollen is the preferred technique to obtain haploid plantlets whose chromosomes are then doubled in Cucurbitaceae, such as melon, cucumber, pumpkin, squash and winter squash. In contrast to doubled haploid procedures in other species, in situ parthenogenesis in cucurbits presents many limiting factors which impede efficient production of haploids. In addition, it is very time-consuming and labor-intensive. However, the haploid inducer-mediated genome-editing system is a breakthrough technology for producing doubled haploids. Several reports have described using the CRISPR/Cas9 system in cucurbit species, and although its application has many bottlenecks, the targeted knock-out of the CENH3 gene will allow breeders to obtain haploid inducer lines that can be used to obtain parthenogenetic embryos. In this review, we discuss the progress made towards the development of doubled haploids and haploid inducer genotypes using CRISPR/Cas9 technologies in cucurbit species. The present review provides insights for the application of haploid inducer-mediated genome-editing system in cucurbit species

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“…Knocking out the function of the PDS gene leads to albinism and slow plant growth [27,28]. As such, PDS has been used as an easy visual phenotypic marker for CRISPR/Cas9-mediated gene knockouts in various plant species, including rice, tobacco, apple, tomato, cassava, banana, wheat, and melon [29][30][31][32][33][34][35][36][37]. Another gene that may provide a visual phenotypic marker for gene knockouts in rice is the young seedling albino (YSA) mutant, which affects the levels of total chlorophyll content in young rice seedlings [38].…”

Section: Introductionmentioning

confidence: 99%

Optimizing Agrobacterium-Mediated Transformation and CRISPR-Cas9 Gene Editing in the tropical japonica Rice Variety Presidio

Molina-Risco

Ibarra

Faion-Molina

et al. 2021

IJMS

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Bottlenecks in plant transformation and regeneration have slowed progress in applying CRISPR/Cas-based genome editing for crop improvement. Rice (Oryza sativa L.) has highly efficient temperate japonica transformation protocols, along with reasonably efficient indica protocols using immature embryos. However, rapid and efficient protocols are not available for transformation and regeneration in tropical japonica varieties, even though they represent the majority of rice production in the U.S. and South America. The current study has optimized a protocol using callus induction from mature seeds with both Agrobacterium-mediated and biolistic transformation of the high-yielding U.S. tropical japonica cultivar Presidio. Gene editing efficiency was tested by evaluating knockout mutations in the phytoene desaturase (PDS) and young seedling albino (YSA) genes, which provide a visible phenotype at the seedling stage for successful knockouts. Using the optimized protocol, transformation of 648 explants with particle bombardment and 532 explants with Agrobacterium led to a 33% regeneration efficiency. The YSA targets had ambiguous phenotypes, but 60% of regenerated plants for PDS showed an albino phenotype. Sanger sequencing of edited progeny showed a number of insertions, deletions, and substitutions at the gRNA target sites. These results pave the way for more efficient gene editing of tropical japonica rice varieties.

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Efficient knockout of phytoene desaturase gene using CRISPR/Cas9 in melon

Cited by 71 publications

References 36 publications

Current Advancements and Limitations of Gene Editing in Orphan Crops

Current Advancements and Limitations of Gene Editing in Orphan Crops

Opportunities and Challenges in Doubled Haploids and Haploid Inducer-Mediated Genome-Editing Systems in Cucurbits

Optimizing Agrobacterium-Mediated Transformation and CRISPR-Cas9 Gene Editing in the tropical japonica Rice Variety Presidio

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