Overexpression of an EaZIP gene devoid of transit peptide sequence induced leaf variegation in tobacco

Guan, Xiayu; Z, Li; Zhang, Zhiliang; Wei, Xiangying; Xie, Jiahua; Chen, Jianjun; Chen, Qing‐Xi

doi:10.1371/journal.pone.0175995

Cited by 3 publications

(5 citation statements)

References 44 publications

(64 reference statements)

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“…The presence and expression of EaZIP were confirmed in all selected seven lines ( Figures 3C,D ). The finding that overexpressing EaZIP was unable to restore the chlorophyll production complements with our previous study that overexpressing the EaZIP caused normal green tobacco plants to become variegated ( Guan et al, 2017 ). The results from these two studies implied that the cloned EaZIP might be defective.…”

Section: Resultssupporting

confidence: 87%

“…Our previous cloning effort obtained an EaZIP from green pothos (Hung et al, 2010), which lacks a sequence region encoding a N-terminal chloroplast transit peptide (cTP). When overexpressing the cloned EaZIP in wild type tobacco plants which carry the NtZIP also lacking 5 -end sequence coding for a cTP (Liu et al, 2004), surprisingly we found transgenic plants showing variegated phenotype (Guan et al, 2017). However, whether the cloned EaZIP is a functional version or not has not been tested.…”

Section: Overexpressing Eazip Did Not Change the Albino Phenotypementioning

confidence: 83%

“…This EaZIP without cTP was initially cloned from green pothos through 5′RACE-PCR techniques ( Hung et al, 2010 ). The expression of this EaZIP in green-leaved tobacco produced variegated transgenic plants ( Guan et al, 2017 ). In addition to this cTP-lacking EaZIP , pothos might have a functional form of EaZIP with sequences encoding the N -terminal cTP.…”

Section: Discussionmentioning

confidence: 99%

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Transformation of Long-Lived Albino Epipremnum aureum ‘Golden Pothos’ and Restoring Chloroplast Development

et al. 2021

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Chloroplasts are organelles responsible for chlorophyll biosynthesis, photosynthesis, and biosynthesis of many metabolites, which are one of key targets for crop improvement. Elucidating and engineering genes involved in chloroplast development are important approaches for studying chloroplast functions as well as developing new crops. In this study, we report a long-lived albino mutant derived from a popular ornamental plant Epipremnum aureum ‘Golden Pothos’ which could be used as a model for analyzing the function of genes involved in chloroplast development and generating colorful plants. Albino mutant plants were isolated from regenerated populations of variegated ‘Golden Pothos’ whose albino phenotype was previously found to be due to impaired expression of EaZIP, encoding Mg-protoporphyrin IX monomethyl ester cyclase. Using petioles of the mutant plants as explants with a traceable sGFP gene, an efficient transformation system was developed. Expressing Arabidopsis CHL27 (a homolog of EaZIP) but not EaZIP in albino plants restored green color and chloroplast development. Interestingly, in addition to the occurrence of plants with solid green color, plants with variegated leaves and pale-yellow leaves were also obtained in the regenerated populations. Nevertheless, our study shows that these long-lived albino plants along with the established efficient transformation system could be used for creating colorful ornamental plants. This system could also potentially be used for investigating physiological processes associated with chlorophyll levels and chloroplast development as well as certain biological activities, which are difficult to achieve using green plants.

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Section: Resultssupporting

confidence: 87%

Section: Overexpressing Eazip Did Not Change the Albino Phenotypementioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

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Transformation of Long-Lived Albino Epipremnum aureum ‘Golden Pothos’ and Restoring Chloroplast Development

et al. 2021

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“…Specifically, mutations leading to variegation, due to their unique feature of exhibiting both normal and defective plastids in different sectors of the same tissue, are of great interest to research toward understanding (1) chloroplast biogenesis, (2) cross-communication between the nucleus and the other DNAcontaining compartments such as plastids and mitochondria, and (3) the molecular mechanism of variegation. Genes involved in the variegation pattern formation have been cloned in monocot and dicot species (Wu et al, 1999;Chen et al, 2000;Takechi et al, 2000;Prikryl et al, 2008;Hayashi-Tsugane et al, 2014;Wang et al, 2016;Zheng et al, 2016;Guan et al, 2017;Zagari et al, 2017). Despite the advances of identifying the underlying genes, insights into the molecular mechanisms controlling the phenomenon are rare (Sakamoto, 2003;Yu et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain Gene in albostrians Barley

Hensel

Mascher

et al. 2019

Plant Cell

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Chloroplasts fuel plant development and growth by converting solar energy into chemical energy. They mature from proplastids through the concerted action of genes in both the organellar and the nuclear genome. Defects in such genes impair chloroplast development and may lead to pigment-deficient seedlings or seedlings with variegated leaves. Such mutants are instrumental as tools for dissecting genetic factors underlying the mechanisms involved in chloroplast biogenesis. Characterization of the green-white variegated albostrians mutant of barley (Hordeum vulgare) has greatly broadened the field of chloroplast biology, including the discovery of retrograde signaling. Here, we report identification of the ALBOSTRIANS gene HvAST (also known as Hordeum vulgare CCT Motif Family gene 7, HvCMF7) by positional cloning as well as its functional validation based on independently induced mutants by Targeting Induced Local Lesions in Genomes (TILLING) and RNA-guided clustered regularly interspaced short palindromic repeats-associated protein 9 endonucleasemediated gene editing. The phenotypes of the independent HvAST mutants imply residual activity of HvCMF7 in the original albostrians allele conferring an imperfect penetrance of the variegated phenotype even at homozygous state of the mutation. HvCMF7 is a homolog of the Arabidopsis (Arabidopsis thaliana) CONSTANS, CO-like, and TOC1 (CCT) Motif transcription factor gene CHLOROPLAST IMPORT APPARATUS2, which was reported to be involved in the expression of nuclear genes essential for chloroplast biogenesis. Notably, in barley we localized HvCMF7 to the chloroplast, without any clear evidence for nuclear localization.

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“…Genes involved in the variegation pattern formation have been cloned in monocot and dicot species (e.g. Wu et al, 1999; Chen et al, 2000; Takechi et al, 2000; Prikryl et al, 2008; Hayashi-Tsugane et al, 2014; Wang et al, 2016; Zheng et al, 2016; Guan et al, 2017; Zagari et al, 2017). Despite the advances of identifying the underlying genes, insights to the molecular mechanisms controlling the phenomenon are rare (Sakamoto, 2003; Yu et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain gene inalbostriansBarley

Hensel

Mascher

et al. 2019

Preprint

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Chloroplasts fuel plant development and growth by converting solar into chemical energy. They mature from proplastids through the concerted action of genes in both the organellar and the nuclear genome. Defects in such genes impair chloroplast development and may lead to pigment-deficient seedlings or seedlings with variegated leaves. Such mutants are instrumental as tools for dissecting genetic factors underlying the mechanisms involved in chloroplast biogenesis. Characterization of the green-white variegated albostrians mutant of barley has greatly broadened the field of chloroplast biology including the discovery of retrograde signaling. Here, we report the identification of the ALBOSTRIANS gene HvAST by positional cloning as well as its functional validation based on independently induced mutants by TILLING and RNA-guided Cas9 endonuclease mediated gene editing. The phenotypes of the independent HvAST mutants imply residual activity of HvAST in the original albostrians allele conferring an imperfect penetrance of the variegated phenotype even at homozygous state of the mutation. HvAST is a homolog of the Arabidopsis thaliana CCT Motif transcription factor gene AtCIA2, which was reported to be involved in the expression of nuclear genes essential for chloroplast biogenesis. Interestingly, in barley we localized HvAST to the chloroplast indicating novel without any clear evidence of nuclear localization.One-sentence summaryLeaf variegation in the barley mutant albostrians is caused by mutation of a single CCT-domain containing gene with residual activity, which is directed to the chloroplast.

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Overexpression of an EaZIP gene devoid of transit peptide sequence induced leaf variegation in tobacco

Cited by 3 publications

References 44 publications

Transformation of Long-Lived Albino Epipremnum aureum ‘Golden Pothos’ and Restoring Chloroplast Development

Transformation of Long-Lived Albino Epipremnum aureum ‘Golden Pothos’ and Restoring Chloroplast Development

Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain Gene in albostrians Barley

Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain gene inalbostriansBarley

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