Efficient production of saffron crocins and picrocrocin in Nicotiana benthamiana using a virus-driven system

Martí, Maricarmen; Diretto, Gianfranco; Aragonés, Verónica; Frusciante, Sarah; Ahrazem, Oussama; Gómez-Gómez, Lourdes; Daròs, José‐Antonio

doi:10.1016/j.ymben.2020.06.009

Cited by 59 publications

(84 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…N. benthamiana wild-type and transformed plants expressing tobacco etch virus (TEV, genus Potyvirus ) nuclear inclusion b (NIb) protein (Martí et al ., 2020), were grown at 25°C under a 12 h/12 h day/night photoperiod. Plants that were 4-to 6-weeks-old were agroinoculated as previously described (Bedoya et al ., 2010; Uranga et al ., 2021).…”

Section: Methodsmentioning

confidence: 99%

CRISPR-Cas12a genome editing at the whole-plant level using two compatible RNA virus vectors

Uranga

Vázquez‐Vilar

Orzáez

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

The use of viral vectors that can replicate and move systemically through the host plant to deliver bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR) components enables genome editing at the whole-plant level and avoids the requirement for labor-intensive stable transformation. However, this approach usually relies on previously transformed plants that stably express a CRISPR-associated (Cas) nuclease. Here we describe successful DNA-free genome editing of Nicotiana benthamiana using two compatible RNA virus vectors, derived from tobacco etch virus (TEV; genus Potyvirus) and potato virus X (PVX; genus Potexvirus), which replicate in the same cells. The TEV and PVX vectors respectively express a Cas12a nuclease and the corresponding guide RNA. This novel two-virus vector system improves the toolbox for transformation-free virus-induced genome editing in plants and will advance efforts to breed more nutritious, resistant, and productive crops.

show abstract

Section: Methodsmentioning

confidence: 99%

CRISPR-Cas12a genome editing at the whole-plant level using two compatible RNA virus vectors

Uranga

Vázquez‐Vilar

Orzáez

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…According to the existing reports ( Table 2 ), many microbes have been successfully transformed to synthesize crocetin or crocins, including E. coli [39] , [44] , Chlorella vulgaris [61] , and yeast [62] . Recently, the transient transformation of Nicotiana benthamiana to synthesize crocins were reported [63] .…”

Section: Crocin Metabolic Engineeringmentioning

confidence: 99%

“…In C. vulgaris , Lou et al transformed crtRB and ZCD1 into a wild type strain with a similar design as above, and produced crocetin without exogenous ALD by using the rich secondary metabolites and potential aldehyde oxidase activity in C. vulgaris [61] . Recently, Martí et al [63] constructed transient transformation vectors for Nicotiana tabacum by using the Tobacco etch virus (TEV), and transformed the tobacco plants with a series of CCD genes and several carotenoid synthesis related genes. The accumulation of yellow pigments of crocins and picrocrocin can be seen with naked eyes in leaves infected by the virus.…”

Section: Crocin Metabolic Engineeringmentioning

confidence: 99%

“…Crocins (0.2%) and picrocrocin (0.8%) were detected in the plants expressing only the CCD gene, which were in relatively low levels. Meanwhile, expression of the phytoene synthase gene ( PaCrtB ) could increase the crocin content, reaching as much as 0.35% of dry weight [63] . This is a successful case of crocin synthesis in higher plants.…”

Section: Crocin Metabolic Engineeringmentioning

confidence: 99%

See 1 more Smart Citation

Prospects and progress on crocin biosynthetic pathway and metabolic engineering

Liu

et al. 2020

Computational and Structural Biotechnology Journal

View full text Add to dashboard Cite

Crocins are a group of highly valuable apocarotenoid-derived pigments mainly produced in Crocus sativus stigmas and Gardenia jasminoides fruits, which display great pharmacological activities for human health, such as anticancer, reducing the risk of atherosclerosis, and preventing Alzheimer’s disease. However, traditional sources of crocins are no longer sufficient to meet current demands. The recent clarification of the crocin biosynthetic pathway opens up the possibility of large-scale production of crocins by synthetic metabolic engineering methods. In this review, we mainly introduce the crocin biosynthetic pathway, subcellular route, related key enzymes, and its synthetic metabolic engineering, as well as its challenges and prospects, with a view to providing useful references for further studies on the synthetic metabolic engineering of crocins.

show abstract

“…Similarly, virus-based expression of Pantoea ananatis phytoene synthase (crtB), the enzyme catalyzing the first step of carotenoid biosynthesis in this soil bacterium, has been demonstrated to lead to a substantial accumulation of phytoene and other carotenoids in plant tissues 5,6 . Virus-based co-expression of crtB and a Crocus sativus carotenoid cleavage dioxygenase (CCD2L) induced large accumulation in N. benthamiana leaves of the apocarotenoid crocins and picrocrocin, which naturally accumulate in saffron stigma and are main constituents of the valued spice 7 . However, these achievements were mainly produced in model plants, such as Nicotiana tabacum , N. benthamiana or Arabidopsis thaliana .…”

Section: Introductionmentioning

confidence: 99%

Carotenoid and tocopherol fortification of zucchini fruits using a viral RNA vector

Houhou

Cordero

Aragonés

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Carotenoids and tocopherols are health-promoting metabolites in livestock and human diets. Some important crops have been genetically modified to increase their content. Although the usefulness of transgenic plants to alleviate nutritional deficiencies is obvious, their social acceptance has been controversial. Here, we demonstrate an alternative biotechnological strategy for carotenoid and tocopherol fortification of edible fruits in which no transgenic DNA is involved. A viral RNA vector derived from Zucchini yellow mosaic virus (ZYMV) was modified to express a bacterial phytoene synthase (crtB), and inoculated in zucchini (Cucurbita pepo L.) leaves nurturing pollinated flowers. After the viral vector moved to the developing fruit and expressed crtB, the rind and flesh of the fruits developed yellow-orange rather than green color. Metabolite analyses showed a substantial enrichment in health-promoting carotenoids, such as α- and β-carotene (pro-vitamin A), lutein and phytoene, in both rind and flesh. Considerably higher accumulation of α- and γ-tocopherol was also detected, particularly in fruit rind. Although this strategy is perhaps not free from controversy due to the use of genetically modified viral RNA, our work does demonstrate the possibility of metabolically fortifying edible fruits using an approach in which no transgenes are involved.

show abstract

Efficient production of saffron crocins and picrocrocin in Nicotiana benthamiana using a virus-driven system

Cited by 59 publications

References 87 publications

CRISPR-Cas12a genome editing at the whole-plant level using two compatible RNA virus vectors

CRISPR-Cas12a genome editing at the whole-plant level using two compatible RNA virus vectors

Prospects and progress on crocin biosynthetic pathway and metabolic engineering

Carotenoid and tocopherol fortification of zucchini fruits using a viral RNA vector

Contact Info

Product

Resources

About