scholarly journals Flower Color Diversity Revealed by Differential Expression of Flavonoid Biosynthetic Genes in Sacred Lotus

2016 ◽  
Vol 141 (6) ◽  
pp. 573-582 ◽  
Author(s):  
Yanjie Wang ◽  
Yeqing Chen ◽  
Man Yuan ◽  
Zeyun Xue ◽  
Qijiang Jin ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Early Stage ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Flower Color ◽  
Flavonoid Biosynthesis ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Biosynthetic Genes ◽  
Sacred Lotus

Sacred lotus (Nelumbo nucifera) is an important aquatic ornamental plant which contains several diverse flower colors, but the underlying mechanisms of its flower coloration remain unclear. In this study, seven complementary DNA (cDNA) clones of genes involved in flavonoid biosynthesis, including chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), flavonoid 3′,5′-hydroxylase (F3′5′H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS), were isolated and characterized. Moreover, expression patterns of these seven genes and pigment profiles were investigated across four N. nucifera cultivars with different flower colors: Zhongguohongbeijing [ZGH (red)], Xinghuafen [XHF (pink)], Molingqiuse [MLQS (yellow)], and Zhufengcuiying [ZFCY (white)]. Real-time quantitative polymerase chain reaction (qRT-PCR) analysis showed that during flower development, transcripts of early biosynthetic genes (NnCHS, NnCHI, and NnF3H) were abundant at the early stage; noticeably, highest expression of NnCHI in MLQS probably induced abundant anthoxanthin synthesis and displayed yellow. Expression of late biosynthetic genes, especially NnDFR and NnANS, was generally consistent with change patterns of anthocyanins in ZGH and XHF, but NnF3′H was barely detectable in the pink cultivars. Meanwhile, negligible expression of NnDFR and NnANS was detected in MLQS and ZFCY, respectively, which blocked their colored anthocyanin biosynthesis. Spatial expression analysis revealed that most flavonoid biosynthetic genes were highly expressed in floral tissues, rather than leaves. These results suggest that in N. nucifera cultivars with different flower colors, flavonoid biosynthesis is differentially regulated by the expression of these flavonoid biosynthetic genes, among which, NnCHI, NnF3′H, NnDFR, and NnANS are supposed to be critical for pigment accumulation, and therefore, affect different flower coloration.

scholarly journals Identification and Functional Analysis of a Flavonol Synthase Gene from Grape Hyacinth

Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1579 ◽  
Author(s):  
Hongli Liu ◽  
Beibei Su ◽  
Han Zhang ◽  
Jiaxin Gong ◽  
Boxiao Zhang ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Flower Color ◽  
Pcr Analysis ◽  
Anthocyanin Content ◽  
In Planta ◽  
Total Anthocyanin ◽  
Flavonol Synthase ◽  
Total Anthocyanin Content ◽  
Grape Hyacinth

Flavonols are important copigments that affect flower petal coloration. Flavonol synthase (FLS) catalyzes the conversion of dihydroflavonols to flavonols. In this study, we identified a FLS gene, MaFLS, expressed in petals of the ornamental monocot Muscari aucheri (grape hyacinth) and analyzed its spatial and temporal expression patterns. qRT-PCR analysis showed that MaFLS was predominantly expressed in the early stages of flower development. We next analyzed the in planta functions of MaFLS. Heterologous expression of MaFLS in Nicotiana tabacum (tobacco) resulted in a reduction in pigmentation in the petals, substantially inhibiting the expression of endogenous tobacco genes involved in anthocyanin biosynthesis (i.e., NtDFR, NtANS, and NtAN2) and upregulating the expression of NtFLS. The total anthocyanin content in the petals of the transformed tobacco plants was dramatically reduced, whereas the total flavonol content was increased. Our study suggests that MaFLS plays a key role in flavonol biosynthesis and flower coloration in grape hyacinth. Moreover, MaFLS may represent a new potential gene for molecular breeding of flower color modification and provide a basis for analyzing the effects of copigmentation on flower coloration in grape hyacinth.

scholarly journals iTRAQ-Based Protein Profiling Provides Insights into the Mechanism of Light-Induced Anthocyanin Biosynthesis in Chrysanthemum (Chrysanthemum × morifolium)

Genes ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 1024
Author(s):  
Yan Hong ◽  
Mengling Li ◽  
Silan Dai
Keyword(s):  
Biosynthetic Pathway ◽  
Developmental Stages ◽  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Flower Color ◽  
Chrysanthemum Morifolium ◽  
Light Signal ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Anthocyanin Biosynthetic Pathway

The generation of chrysanthemum (Chrysanthemum × morifolium) flower color is mainly attributed to the accumulation of anthocyanins. Light is one of the key environmental factors that affect the anthocyanin biosynthesis, but the deep molecular mechanism remains elusive. In our previous study, a series of light-induced structural and regulatory genes involved in the anthocyanin biosynthetic pathway in the chrysanthemum were identified using RNA sequencing. In the present study, differentially expressed proteins that are in response to light with the capitulum development of the chrysanthemum ‘Purple Reagan’ were further identified using isobaric tags for relative and absolute quantification (iTRAQ) technique, and correlation between the proteomic and the transcriptomic libraries was analyzed. In general, 5106 raw proteins were assembled based on six proteomic libraries (three capitulum developmental stages × two light treatments). As many as 160 proteins were differentially expressed between the light and the dark libraries with 45 upregulated and 115 downregulated proteins in response to shading. Comparative analysis between the pathway enrichment and the gene expression patterns indicated that most of the proteins involved in the anthocyanin biosynthetic pathway were downregulated after shading, which was consistent with the expression patterns of corresponding encoding genes; while five light-harvesting chlorophyll a/b-binding proteins were initially downregulated after shading, and their expressions were enhanced with the capitulum development thereafter. As revealed by correlation analysis between the proteomic and the transcriptomic libraries, GDSL esterase APG might also play an important role in light signal transduction. Finally, a putative mechanism of light-induced anthocyanin biosynthesis in the chrysanthemum was proposed. This study will help us to clearly identify light-induced proteins associated with flower color in the chrysanthemum and to enrich the complex mechanism of anthocyanin biosynthesis for use in cultivar breeding.

scholarly journals Isolation, Characterization, and Expression Analysis of the MaMDH Gene in Banana

HortScience ◽  
2013 ◽  
Vol 48 (5) ◽  
pp. 614-619
Author(s):  
Cai-Hong Jia ◽  
Ju-Hua Liu ◽  
Zhi-Qiang Jin ◽  
Qiu-Ju Deng ◽  
Jian-Bin Zhang ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Ethylene Biosynthesis ◽  
Pcr Analysis ◽  
Malic Dehydrogenase ◽  
Reading Frame ◽  
Expression Levels ◽  
Post Harvest ◽  
Real Time Quantitative Pcr

A full-length cDNA isolated from banana (Musa acuminata L. AAA group) fruit was named MaMDH, containing an open reading frame encoding 332 amino acids that represents the gene for cytoplasmic malic dehydrogenase (MDH). Sequence analysis showed that MaMDH shares high similarity with MDHs from castor bean (XP_002533463), tobacco (CAC12826), peach (AAL11502), and chickpeas (CAC10208). Real-time quantitative polymerase chain reaction (PCR) analysis of MaMDH spatial expression showed that it was expressed in all organs examined: roots, rhizomes, leaves, flowers, and fruits. The expression was the highest in flowers followed by the fruits and roots, whereas the rhizomes and leaves displayed the lowest expression levels. Real-time quantitative PCR revealed that MaMDH exhibited differential expression patterns in post-harvest banana fruits correlating with ethylene biosynthesis. In naturally ripened banana fruits, MaMDH expression was in accordance with ethylene biosynthesis. In accordance, for banana fruits treated with the ethylene analog 1-methylclopropene (1-MCP), MaMDH expression levels were inhibited and remained constant. After treatment with ethylene, MaMDH expression in banana fruits significantly increased with ethylene biosynthesis and peaked 3 days after harvest, which was 11 days earlier than that in naturally ripened banana fruits. These results suggest that MaMDH expression is induced by ethylene to regulate post-harvest banana fruits ripening.

scholarly journals Comparative transcriptome analysis of inbred lines and contrasting hybrids reveals overdominance mediate early biomass vigor in hybrid cotton

2020 ◽  
Author(s):  
Kashif Shahzad ◽  
Xuexian Zhang ◽  
Liping Guo ◽  
Tingxiang Qi ◽  
Huini Tang ◽  
...  
Keyword(s):  
Circadian Rhythm ◽  
Transcriptome Analysis ◽  
Molecular Mechanisms ◽  
Early Stage ◽  
Expression Patterns ◽  
Pcr Analysis ◽  
Genetic Constitution ◽  
Comparative Transcriptome ◽  
Comparative Transcriptome Analysis ◽  
Extracellular Region

Abstract Background: Heterosis breeding is the most useful method for yield increase around the globe. Heterosis is an intriguing process to develop superior offspring to either parent in the desired character. The biomass vigor produced during seedling emergence stage has a direct influence on yield heterosis in plants. Unfortunately, the genetic basis of early biomass vigor in cotton is poorly understood. Results: Three stable performing F1 hybrids varying in yield heterosis named as high, medium and low hybrids with their inbred parents were used in this study. Phenotypically, these hybrids established noticeable biomass heterosis during the early stage of seedling growth in the field. Transcriptome analysis of root and leaf revealed that hybrids showed many differentially expressed genes (DEGs) relative to their parents, while the comparison of inbred parents showed limited number of DEGs indicating similarity in their genetic constitution. Further analysis indicated expression patterns of most DEGs were overdominant in both tissues of hybrids. According to GO results, functions of overdominance genes in leaf were enriched for chloroplast, membrane, and protein binding, whereas functions of overdominance genes in root were enriched for plasma membrane, extracellular region, and responses to stress. We found several genes of circadian rhythm pathway related to LATE ELONGATED HYPOCOTYL (LHY) showed downregulated overdominant expressions in both tissues of hybrids. In addition to circadian rhythm, several leaf genes related to Aux/IAA regulation, and many root genes involved in peroxidase activity also showed overdominant expressions in hybrids. Twelve genes involved in circadian rhythm plant were selected to perform qRT-PCR analysis to confirm the accuracy of RNA-seq results. Conclusions: Through genome-wide comparative transcriptome analysis, we strongly predict that overdominance at gene expression level plays a pivotal role in early biomass vigor of hybrids. The combinational contribution of circadian rhythm and other metabolic process may control vigorous growth in hybrids. Our result provides an important foundation for dissecting molecular mechanisms of biomass vigor in hybrid cotton.

scholarly journals Expression of Genes Involved in Proanthocyanidin Biosynthesis during Fruit Development in a Chinese Pollination-constant, Nonastringent (PCNA) Persimmon, `Luo Tian Tian Shi'

2005 ◽  
Vol 130 (6) ◽  
pp. 830-835 ◽  
Author(s):  
Ayako Ikegami ◽  
Keizo Yonemori ◽  
Akira Kitajima ◽  
Akihiko Sato ◽  
Masahiko Yamada
Keyword(s):  
Fruit Development ◽  
Early Stage ◽  
Condensed Tannin ◽  
Expression Patterns ◽  
Subtractive Hybridization ◽  
Cell Development ◽  
Flavonoid Biosynthesis ◽  
Fruit Growth ◽  
Diospyros Kaki ◽  
Serine Carboxypeptidase

Expression patterns of the genes involved in condensed tannin (CT) biosynthesis during fruit development was investigated in a Chinese pollination-constant, nonastringent (PCNA) persimmon (Diospyros kaki Thunb.) `Luo Tian Tian Shi'. The transcript levels of phenylalanine ammonia-lyase (PAL) and dihydroflavonol reductase (DFR) in `Luo Tian Tian Shi' were detected at high levels throughout the fruit growth. Chalcone synthase (CHS) and flavonol 3-hydroxylase (F3H) also continued to be transcribed during fruit growth, although their levels decreased earlier than PAL and DFR. In contrast, expression levels of these genes declined into undetectable levels at an early stage of fruit development in Japanese PCNA persimmon. In addition, anthocyanidin reductase (ANR), which encodes a key enzyme of the proanthocyanidin biosynthesis, was transcribed at high levels in `Luo Tian Tian Shi' during fruit growth, but not in Japanese PCNA persimmon. By contrast, the expression of D. kaki serine carboxypeptidase-like protein 1 (DkSCPL1) that was obtained from suppression subtractive hybridization (SSH) analysis between artificially astringency-removed fruit and astringent fruit in a different experiment, declined earlier than the other flavonoid biosynthesis genes in `Luo Tian Tian Shi', coincident with the termination of the tannin cell development. In the F1 progeny of the cross between `Luo Tian Tian Shi' and Japanese PCNA `Taishu', similar expression patterns were obtained among segregated PCNA and astringent offspring. These results indicate that Chinese PCNA is different from Japanese PCNA in expression of the genes involved in CT biosynthesis. In conclusion, we clarified that expression of the genes (PAL to ANR, but not SCPL) involved in flavonoid biosynthesis was continuous in the Chinese PCNA cultivar, despite the termination of tannin cell development.

scholarly journals Genome-Wide Analysis of MYB Gene Family in Chinese Bayberry (Morella rubra) and Identification of Members Regulating Flavonoid Biosynthesis

2021 ◽  
Vol 12 ◽  
Author(s):  
Yunlin Cao ◽  
Huimin Jia ◽  
Mengyun Xing ◽  
Rong Jin ◽  
Donald Grierson ◽  
...  
Keyword(s):  
Gene Family ◽  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Flavonoid Biosynthesis ◽  
Structure Evolution ◽  
Chinese Bayberry ◽  
Genome Wide Analysis ◽  
Myb Gene ◽  
Genome Wide ◽  
Family Analysis

Chinese bayberry (Morella rubra), the most economically important fruit tree in the Myricaceae family, is a rich source of natural flavonoids. Recently the Chinese bayberry genome has been sequenced, and this provides an opportunity to investigate the organization and evolutionary characteristics of MrMYB genes from a whole genome view. In the present study, we performed the genome-wide analysis of MYB genes in Chinese bayberry and identified 174 MrMYB transcription factors (TFs), including 122 R2R3-MYBs, 43 1R-MYBs, two 3R-MYBs, one 4R-MYB, and six atypical MYBs. Collinearity analysis indicated that both syntenic and tandem duplications contributed to expansion of the MrMYB gene family. Analysis of transcript levels revealed the distinct expression patterns of different MrMYB genes, and those which may play important roles in leaf and flower development. Through phylogenetic analysis and correlation analyses, nine MrMYB TFs were selected as candidates regulating flavonoid biosynthesis. By using dual-luciferase assays, MrMYB12 was shown to trans-activate the MrFLS1 promoter, and MrMYB39 and MrMYB58a trans-activated the MrLAR1 promoter. In addition, overexpression of 35S:MrMYB12 caused a significant increase in flavonol contents and induced the expression of NtCHS, NtF3H, and NtFLS in transgenic tobacco leaves and flowers and significantly reduced anthocyanin accumulation, resulting in pale-pink or pure white flowers. This indicates that MrMYB12 redirected the flux away from anthocyanin biosynthesis resulting in higher flavonol content. The present study provides valuable information for understanding the classification, gene and motif structure, evolution and predicted functions of the MrMYB gene family and identifies MYBs regulating different aspects of flavonoid biosynthesis in Chinese bayberry.

Structures and expression patterns of two cDNA clones encoding S-adenosyl-L-methionine synthetase from the root nodule of Elaeagnus umbellata

2001 ◽  
Vol 28 (9) ◽  
pp. 951
Author(s):  
Sang Ho Lee ◽  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acid ◽  
Expression Pattern ◽  
Root Nodule ◽  
Vascular System ◽  
Expression Patterns ◽  
Amino Acid Level ◽  
Nodule Development ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Elaeagnus Umbellata

This paper originates from an address at the 8th International Symposium on Nitrogen Fixation with Non-Legumes, Sydney, NSW, December 2000 Two cDNA clones encoding S-adenosyl-L-methionine synthetase (SAMS) were isolated from the root nodule cDNA library of Elaeagnus umbellata Thunberg and analysed on the basis of deduced amino acid sequence and expression pattern. Two EuSAMS clones shared 75–84% identity at the nucleotide level, and 85–95% identity at the amino acid level, with the other plant SAMS genes. Genomic Southern hybridization revealed the presence of more than two copies of SAMS genes in the genome of E. umbellata. Reverse transcriptase-mediated polymerase chain reaction (RT–PCR) analysis showed EuSAMS1 transcripts were more abundant than those of EuSAMS2. Similar to the expression pattern of other plant SAMS genes, both genes were expressed at higher levels in root than in leaf. During nodule development, expression of both genes was increased, with the highest level at 6–8 week after inoculation, and decreased rapidly thereafter. In situ hybridization analysis also showed both SAMS transcripts in the meristem zone, the infected cells of the fixation zone and in the central vascular system of root nodules. However, EuSAMS2 transcripts were strongly detected in the prefixation zone, whereas EuSAMS1 transcripts were hardly detected. These results suggest different regulatory mechanisms for the two genes in the root nodule. The expression pattern of SAMS genes in the root nodule may correlate mostly with cell wall synthesis, polyamine biosynthesis and other methylation-mediated functions.

scholarly journals Mechanism Underlying Color Variation in Calla Lily Spathes Based on Transcriptomic Analysis

2021 ◽  
pp. 1-16
Author(s):  
Ying Fang ◽  
Ting Lei ◽  
Yanmei Wu ◽  
Xuehua Jin
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Flower Color ◽  
Transcriptomic Analysis ◽  
Color Variation ◽  
Anthocyanin Synthesis ◽  
Helix Loop Helix ◽  
Calla Lily ◽  
Unique Shape ◽  
Glutathione S Transferases

The calla lily (Zantedeschia hybrida) is a valued ornamental plant due to its unique shape and color variations. To determine the mechanisms responsible for color development in the calla lily spathe, we conducted a comparative transcriptomic analysis of the spathes of the black [Black Girl (B)], pink [Romantic (P)], and white [Ventura (W)] cultivars. The gene expression patterns in six spathe colors, including the preceding three colors as well as the amaranth [Promise (N)], red [Figo (F)], and yellow [Sun Club (Y)] cultivars were analyzed by real-time quantitative polymerase chain reaction (PCR). Transcriptomic analysis identified 25,165 differentially expressed genes. The transcription abundance and expression level of genes annotated as anthocyanidin reductase (ANR1, ANR2), basic-helix-loop-helix (bHLH1), and glutathione S-transferases (GST1) were significantly upregulated in B, and the expression of anthocyanidin synthase (ANS) was highest in B except for N. However, chalcone isomerase (CHI2) and dihydroflavonol 4-reductase (DFR1, DFR2) were expressed at significantly lower levels in P, W, and Y. Correlation analysis revealed that bHLH1 might act as a positive regulator of ANS expression, promoting anthocyanin synthesis. Moreover, GST1-encoded proteins may be related to the accumulation and transport of both anthocyanin and procyanidin in the calla lily spathe. It is speculated that the formation of the black spathe is related to the accumulation of anthocyanins and procyanidins. However, the low expression of CHI2, DFR1, and DFR2 may result in the inhibition of anthocyanin synthesis, which may lead to lightening of the spathe color. This preliminary study revealed the mechanism responsible for calla lily spathe color, identifying the key genes involved, thus providing effective gene resources and a theoretical basis for flower color molecular breeding.

scholarly journals REVEILLE Transcription Factors Contribute to the Nighttime Accumulation of Anthocyanins in ‘Red Zaosu’ (Pyrus Bretschneideri Rehd.) Pear Fruit Skin

2020 ◽  
Vol 21 (5) ◽  
pp. 1634
Author(s):  
Xieyu Li ◽  
Ting Wu ◽  
Hanting Liu ◽  
Rui Zhai ◽  
Yao Wen ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Luciferase Reporter ◽  
Anthocyanin Accumulation ◽  
Pear Fruit ◽  
Biosynthetic Genes ◽  
Expression Levels ◽  
Fruit Skin ◽  
Genes Encoding

Anthocyanin biosynthesis exhibits a rhythmic oscillation pattern in some plants. To investigate the correlation between the oscillatory regulatory network and anthocyanin biosynthesis in pear, the anthocyanin accumulation and the expression patterns of anthocyanin late biosynthetic genes (ALBGs) were investigated in fruit skin of ‘Red Zaosu’ (Pyrus bretschneideri Rehd.). The anthocyanin accumulated mainly during the night over three continuous days in the fruit skin, and the ALBGs’ expression patterns in ‘Red Zaosu’ fruit skin were oscillatory. However, the expression levels of typical anthocyanin-related transcription factors did not follow this pattern. Here, we found that the expression patterns of four PbREVEILLEs (PbRVEs), members of a class of atypical anthocyanin-regulated MYBs, were consistent with those of ALBGs in ‘Red Zaosu’ fruit skin over three continuous days. Additionally, transient expression assays indicated that the four PbRVEs promoted anthocyanin biosynthesis by regulating the expression of the anthocyanin biosynthetic genes encoding dihydroflavonol-4-reductase (DFR) and anthocyanidin synthase (ANS) in red pear fruit skin, which was verified using a dual-luciferase reporter assay. Moreover, a yeast one-hybrid assay indicated that PbRVE1a, 1b and 7 directly bound to PbDFR and PbANS promoters. Thus, PbRVEs promote anthocyanin accumulation at night by up-regulating the expression levels of PbDFR and PbANS in ‘Red Zaosu’ fruit skin.

scholarly journals Anthocyanin Accumulation and Related Gene Expression Profile in ‘Red Zaosu’ Pear and Its Green Mutant

Agriculture ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 898
Author(s):  
Yunting Zhang ◽  
Shanlin Li ◽  
Xianjie Gu ◽  
Diya Lei ◽  
Bing Zhao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Anthocyanin Biosynthesis ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Biosynthetic Genes ◽  
Specific Expression ◽  
Anthocyanin Biosynthetic Genes ◽  
Green Mutant

Red-skinned pear is a promising commercial fruit due to its attractive appearance and nutritious value. Anthocyanin is the determinant of the red coloration of the pear peel. However, differences in anthocyanin accumulation exist among red pear cultivars with different genetic backgrounds. In this study, we analyzed the anthocyanin content and gene expression patterns in the fruits and different tissues of the red pear ‘Red Zaosu’ at different developmental stages and found a difference in anthocyanin accumulation between ‘Red Zaosu’ pear and its green mutant. The data showed that the expression profiles of transcripts that encoded critical anthocyanin biosynthetic genes were basically consistent with a tendency to a decreased anthocyanin content during fruit development, indicating that a synergistic effect of these genes was responsible for anthocyanin biosynthesis and regulation. Tissue-specific expression analysis of anthocyanin biosynthetic genes showed that they could be expressed in all tissues but at different levels. PbF3H, PbDFR, and PbANS were mainly expressed during the early flowering period, which explained the reduced levels of anthocyanin content in petals. Additionally, the content of anthocyanins and the expression levels of PbDFR, PbANS, and PbMYB10 significantly decreased in the green mutant of ‘Red Zaosu’, suggesting that PbDFR, PbANS, and PbMYB10 probably play a decisive role in determining the skin coloration of ‘Red Zaosu’ and its green mutant.

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