Color-related chlorophyll and carotenoid concentrations of Chinese kale can be altered through CRISPR/Cas9 targeted editing of the carotenoid isomerase gene BoaCRTISO

Sun, Bo; Jiang, Min; Zheng, Hao; Jian, Yue; Huang, Wenli; Qiao, Yuan; Zheng, Aihong; Chen, Qing; Zhang, Yunting; Lin, Yuanxiu; Wang, Yan; Wang, Xiaorong; Wang, Qiaomei; Zhang, Fen; Tang, Haoru

doi:10.1038/s41438-020-00379-w

Cited by 57 publications

(40 citation statements)

References 38 publications

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“…Moreover, we found three ISO genes ( BnZ-ISOa , BnZ-ISOa and BnCrtISOa ) were significantly downregulated in WP vs. ZS11 petals at S1, which may be related to the reduced total lutein and zeaxanthin contents in the white-flowered line. In the crtiso mutants of Chinese kale, the carotenoid isomerase gene ( BoaCRTISO ) was targeted and edited using the CRISPR/Cas9 system, the expression levels of CRTISO and most carotenoid and chlorophyll biosynthesis-related genes were notably lower than in the WT plants, in addition, both the total and individual carotenoid and chlorophyll concentrations were reduced, and the total levels declined by 11.89–36.33% (Sun et al 2020 ). Most importantly, BnNCED4b was dramatically upregulated in WP petals and showed almost no expression in ZS11 petals, suggesting that this gene plays a key role in turning WP petals from pale yellow to white.…”

Section: Discussionmentioning

confidence: 99%

Comparative transcriptomic and metabolomic analyses of carotenoid biosynthesis reveal the basis of white petal color in Brassica napus

Jia

Wang

et al. 2021

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Main conclusion The molecular mechanism underlying white petal color in Brassica napus was revealed by transcriptomic and metabolomic analyses. Abstract Rapeseed (Brassica napus L.) is one of the most important oilseed crops worldwide, but the mechanisms underlying flower color in this crop are known less. Here, we performed metabolomic and transcriptomic analyses of the yellow-flowered rapeseed cultivar ‘Zhongshuang 11’ (ZS11) and the white-flowered inbred line ‘White Petal’ (WP). The total carotenoid contents were 1.778-fold and 1.969-fold higher in ZS11 vs. WP petals at stages S2 and S4, respectively. Our findings suggest that white petal color in WP flowers is primarily due to decreased lutein and zeaxanthin contents. Transcriptome analysis revealed 10,116 differentially expressed genes with a fourfold or greater change in expression (P-value less than 0.001) in WP vs. ZS11 petals, including 1,209 genes that were differentially expressed at four different stages and 20 genes in the carotenoid metabolism pathway. BnNCED4b, encoding a protein involved in carotenoid degradation, was expressed at abnormally high levels in WP petals, suggesting it might play a key role in white petal formation. The results of qRT-PCR were consistent with the transcriptome data. The results of this study provide important insights into the molecular mechanisms of the carotenoid metabolic pathway in rapeseed petals, and the candidate genes identified in this study provide a resource for the creation of new B. napus germplasms with different petal colors.

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Section: Discussionmentioning

confidence: 99%

Comparative transcriptomic and metabolomic analyses of carotenoid biosynthesis reveal the basis of white petal color in Brassica napus

Jia

Wang

et al. 2021

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“…Therefore, the chlorophyll content showed a higher decrease than the carotenoid content, and the decrease was greater for chlorophyll b content than chlorophyll a content. Previous studies have shown that chlorophyll is responsible for the green pigmentation in plants, and the presence of carotenoids affects leaf color (Wu et al, 2018 ; Sun et al, 2020 ). We speculated that this significant change in the photosynthetic pigment was the direct cause of the pale green phenotype in pem1 and pem2 .…”

Section: Resultsmentioning

confidence: 99%

Mapping of a Pale Green Mutant Gene and Its Functional Verification by Allelic Mutations in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

et al. 2021

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Leaves are the main organ for photosynthesis, and variations in leaf color affect photosynthesis and plant biomass formation. We created two similar whole-plant pale green mutants (pem1 and pem2) from the double haploid (DH) Chinese cabbage line “FT” through ethyl methanesulfonate (EMS) mutagenesis of seeds. Photosynthetic pigment contents and net photosynthetic rates were significantly lower in the mutants than in the wild-type “FT,” and the chloroplast thylakoid endomembrane system was poor. Genetic analysis showed that the mutated phenotypes of pem1 and pem2 were caused by a single nuclear gene. Allelism tests showed that pem1 and pem2 were alleles. We mapped Brpem1 to a 64.25 kb region on chromosome A10, using BSR-Seq and map-based cloning of 979 F2 recessive individuals. Whole-genome re-sequencing revealed a single nucleotide polymorphism (SNP) transition from guanine to adenosine on BraA10g021490.3C in pem1, causing an amino acid shift from glycine to glutamic acid (G to E); in addition, BraA10g021490.3C in pem2 was found to have a single nucleotide substitution from guanine to adenosine, causing an amino acid change from E to lysine (K). BraA10g021490.3C is a homolog of the Arabidopsisdivinyl chlorophyllide a 8-vinyl-reductase (DVR) gene that encodes 3,8-divinyl protochlorophyllide a 8-vinyl reductase, which is a key enzyme in chlorophyll biosynthesis. Enzyme activity assay and chlorophyll composition analysis demonstrated that impaired DVR had partial loss of function. These results provide a basis to understand chlorophyll metabolism and explore the mechanism of a pale green phenotype in Chinese cabbage.

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“…These findings indicate that the phenotypes of leaves and other organs are expressed differently according to species after CRTISO is silenced. Besides, after the BoaCRTISO gene was target-edited in Chinese kale, the plant showed a distinct yellow color, although the gene expression level was similar to that of the VIGS plants (Sun et al, 2020). This shows that stable plant transformation can make plants show a more obvious phenotype than transient VIGS technology.…”

Section: Discussionmentioning

confidence: 96%

Characterization of BoaCRTISO Reveals Its Role in Carotenoid Biosynthesis in Chinese Kale

et al. 2021

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Carotenoids are organic pigments that play an important role in both plant coloration and human health; they are a critical subject in molecular breeding due to growing demand for natural molecules in both food and medicine. In this study, we focus upon characterizing BoaCRTISO, the carotenoid isomerase gene before the branch of the carotenoid biosynthetic pathway, which is expressed in all organs and developmental stages of Chinese kale, and BoaCRTISO, which is located in the chloroplast. The expression of BoaCRTISO is induced by strong light, red and blue combined light, and gibberellic acid treatment, but it is suppressed by darkness and abscisic acid treatment. We obtained BoaCRTISO-silenced plants via virus-induced gene silencing technology, and the silence efficiencies ranged from 52 to 77%. The expressions of most carotenoid and chlorophyll biosynthetic genes in BoaCRTISO-silenced plants were downregulated, and the contents of carotenoids and chlorophyll were reduced. Meanwhile, BoaCRTISO-silenced plants exhibited phenotypes of yellowing leaves and inhibited growth. This functional characterization of BoaCRTISO provides insight for the biosynthesis and regulation of carotenoid in Chinese kale.

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Color-related chlorophyll and carotenoid concentrations of Chinese kale can be altered through CRISPR/Cas9 targeted editing of the carotenoid isomerase gene BoaCRTISO

Cited by 57 publications

References 38 publications

Comparative transcriptomic and metabolomic analyses of carotenoid biosynthesis reveal the basis of white petal color in Brassica napus

Comparative transcriptomic and metabolomic analyses of carotenoid biosynthesis reveal the basis of white petal color in Brassica napus

Mapping of a Pale Green Mutant Gene and Its Functional Verification by Allelic Mutations in Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

Characterization of BoaCRTISO Reveals Its Role in Carotenoid Biosynthesis in Chinese Kale

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