Leaf Color Mutants Research Articles

Slym1 control the color etiolation of leaves by facilitating the decomposition of chlorophyll in tomato

Photosynthesis, as an important biological process of plants, produces organic substances for plant growth and development. Although the molecular mechanisms of photosynthesis had been well investigated, the relationship between chlorophyll synthesis and photosynthesis remains largely unknown. The leaf-color mutant was an ideal material for studying photosynthesis and chlorophyll synthesis, which had been seldom investigated in tomato. Here, we obtained a yellow leaf tomato mutant ym (The mutant plants from the line of zs4) in field. Transmission electron microscopy (TEM) and photosynthetic parameters results demonstrated that chloroplast’s structure was obviously destroyed and photosynthetic capacity gets weak. The mutant was hybridized with the control to construct the F2 segregation population for sequencing. Slym1 gene, controlling yellow mutant trait, was identified using Bulked Segregation Analysis. Slym1 was up-regulated in the mutant and Slym1 was located in the nucleus. The genes associated with photosynthesis and chlorophyll synthesis were down-regulated in Slym1-OE transgenic tomato plants. The results suggested that Slym1 negatively regulate photosynthesis. Photosynthetic pigment synthesis related genes HEMA, HEMB1, CHLG and CAO were up-regulated in Slym1 silencing plants. The redundant Slym1 binding the intermediate proteins MP resulting in hindering the interaction between MP and HY5 due to the Slym1 with a high expression level in ym mutant, lead to lots of the HY5 with unbound state accumulates in cells, that could accelerate the decomposition of chlorophyll. Therefore, the yellow leaf-color mutant ym could be used as an ideal material for further exploring the relationship between leaf color mutant and photosynthesis and the specific mechanism.

Cia Zeaxanthin Biosynthesis, OsZEP and OsVDE Regulate Striped Leaves Occurring in Response to Deep Transplanting of Rice.

The rice leaf color mutant B03S was previously generated from the photoperiod- and thermo-sensitive genic male sterile (PTGMS) rice line Efeng 1S, of which male sterility manifests by photoperiod and temperature but exhibits mainly temperature-sensitive characteristics. After these plants were deeply transplanted, the new leaves manifested typical zebra stripe patterns. Here, B03S was subjected to deep and shallow transplanting, shading with soil and aluminum foil, and control conditions in situ to determine the cause of the striped-leaf trait. The direct cause of striped leaves is the base of the leaf sheath being under darkness during deep transplanting, of which the critical shading range reached or exceeds 4 cm above the base. Moreover, typical striped leaves were analyzed based on the targeted metabolome method by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC–MS/MS) combined with transcriptome and real-time quantitative PCR (qPCR)-based verification to clarify the metabolic pathways and transcriptional regulation involved. Carotenoids enter the xanthophyll cycle, and the metabolites that differentially accumulate in the striped leaves include zeaxanthin and its derivatives for photooxidative stress protection, driven by the upregulated expression of OsZEP. These findings improve the understanding of the physiological and metabolic mechanisms underlying the leaf color mutation in rice plants, enrich the theoretical foundation of the nonuniform leaf color phenomenon widely found in nature and highlight key advancements concerning rice production involving the transplanting of seedlings or direct broadcasting of seeds.

VG, encoding a thylakoid formation protein, regulates the formation of variegated leaves in tomato

Leaf-color mutations have been studied extensively in plants. However, to better understand the complex mechanisms underlying the formation of leaf color, it is essential to continue discover novel genes involved in the process of leaf color development. In this study, we identified a variegated-leaf (vg) mutant in tomato that exhibited defective phenotypes in thylakoids and photosynthesis. To clone the vg locus, an F2 population was constructed from the cross between the vg mutant (Solanum lycopersicum) and the wild tomato LA1589 (S. pimpinellifolium). Using the map-based cloning approach, the vg locus was mapped on chromosome 7 and narrowed down to a 128 kb region that contained 21 open reading frames (ORFs). The expression levels of ORF9, ORF10, and ORF13 were significantly lower in vg than in the wild-type plants, while the ORF11 transcript level was elevated in vg. We then mutagenized ORF9, ORF10, and ORF13 by the CRISPR/Cas9 system in the wild-type tomato background and found that only the ORF10 mutation reproduced the phenotype of variegated leaves, indicating that ORF10 represents VG and its down-regulated expression was responsible for the variegated leaf phenotype. ORF10 encodes a thylakoid formation protein and its mutant lines showed reduced levels of chlorophyll synthesis and photosynthesis. Taken together, these results suggest that VG is necessary for chloroplast development, chlorophyll synthesis, and photosynthesis in tomato.

Comparative Transcriptome Profiling Analysis Reveals the Adaptive Molecular Mechanism of Yellow-Green Leaf in Rosa beggeriana 'Aurea'.

Rosa beggeriana ‘Aurea’ is a yellow-green leaf (yl) mutant and originated from Rosa beggeriana Schrenk by 60Co-γ irradiation, which is an important ornamental woody species. However, the molecular mechanism of the yl mutant remains unknown. Herein, comparative transcriptome profiling was performed between the yl type and normal green color type (WT) by RNA sequencing. A total of 3,372 significantly differentially expressed genes (DEGs) were identified, consisting of 1,585 upregulated genes and 1,787 downregulated genes. Genes that took part in metabolic of biological process (1,090), membrane of cellular component (728), catalytic (1,114), and binding of molecular function (840) were significantly different in transcription level. DEGs involved in chlorophyll biosynthesis, carotenoids biosynthesis, cutin, suberine, wax biosynthesis, photosynthesis, chloroplast development, photosynthesis-antenna proteins, photosystem I (PSI) and photosystem II (PSII) components, CO2 fixation, ribosomal structure, and biogenesis related genes were downregulated. Meanwhile, linoleic acid metabolism, siroheme biosynthesis, and carbon source of pigments biosynthesis through methylerythritol 4-phosphate (MEP) pathways were upregulated. Moreover, a total of 147 putative transcription factors were signification different expression, involving NAC, WRKY, bHLH, MYB and AP2/ERF, C2H2, GRAS, and bZIP family gene. Our results showed that the disturbed pigments biosynthesis result in yl color by altering the ratio of chlorophylls and carotenoids in yl mutants. The yl mutants may evoke other metabolic pathways to compensate for the photodamage caused by the insufficient structure and function of chloroplasts, such as enhanced MEP pathways and linoleic acid metabolism against oxidative stress. This research can provide a reference for the application of leaf color mutants in the future.

Insights into the molecular basis of a yellow leaf color mutant (ym) in tomato (Solanum lycopersicum)

Leaf color mutants are ideal materials for studying mechanisms of plant such as photosynthesis, photorespiration, the pigment biosynthesis, and some other agronomic traits. Meanwhile, leaf color mutants are widely used in the screening of the offspring of cross breeding. The molecular mechanisms of yellow leaf and REDOX balance of tomatoes have not been explored. Here we obtain a naturally yellow leaf tomato mutant (ym). In order to further understand the role of REDOX, a series of experiments are conducted to assess the physiological and molecular reactions of two tomato lines, ym and zs4. The results showed that the ym mutant cells accumulated excessive H2O2. The chlorophyll content of the mutant decreased greatly. The chloroplast structure of the mutant is observed by transmission electron microscope (TEM). The results showed that mutant's chloroplast membrane structure is seriously damaged. A transcriptomic approach is used to analyze the gene regulate etiolation traits in ym mutant tomato plants. Many genes are enriched in oxidation-reduction (REDOX). Kyoto encyclopedia of genes and genomes (KEGG) revealed that DEGs are involved in pathways such as plant hormone signal transduction, carotenoid biosynthesis, MAPK signaling pathway, phenols and flavonoids synthesis, and pentose and glucuronate interconversions, which further confirmed the role of yellow trait. Overall, the damage of chloroplast structure, low pigment content and low level ROS scavenging capability are the main factors leading to yellow leaf.Main ConclusionDamage of chloroplast structure lead to the low level of the chlorophyll and the REDOX imbalance cause tomato yellow leaf color.

Comparative transcriptome and microbial community sequencing provide insight into yellow-leaf phenotype of Camellia japonica

BackgroundLeaf color variation is a common trait in plants and widely distributed in many plants. In this study, a leaf color mutation in Camellia japonica (cultivar named as Maguxianzi, M) was used as material, and the mechanism of leaf color variation was revealed by physiological, cytological, transcriptome and microbiome analyses.ResultsThe yellowing C. japonica (M) exhibits lower pigment content than its parent (cultivar named as Huafurong, H), especially chlorophyll (Chl) and carotenoid, and leaves of M have weaker photosynthesis. Subsequently, the results of transmission electron microscopy(TEM) exhibited that M chloroplast was accompanied by broken thylakoid membrane, degraded thylakoid grana, and filled with many vesicles. Furthermore, comparative transcriptome sequencing identified 3,298 differentially expressed genes (DEGs). KEGG annotation analysis results showed that 69 significantly enriched DEGs were involved in Chl biosynthesis, carotenoid biosynthesis, photosynthesis, and plant-pathogen interaction. On this basis, we sequenced the microbial diversity of the H and M leaves. The sequencing results suggested that the abundance of Didymella in the M leaves was significantly higher than that in the H leaves, which meant that M leaves might be infected by Didymella.ConclusionsTherefore, we speculated that Didymella infected M leaves while reduced Chl and carotenoid content by damaging chloroplast structures, and altered the intensity of photosynthesis, thereby causing the leaf yellowing phenomenon of C. japonica (M). This research will provide new insights into the leaf color variation mechanism and lay a theoretical foundation for plant breeding and molecular markers.

Comparative transcriptome analysis reveals candidate genes for leaf color formation in a thermo-sensitive leaf-color mutant generated by carbon-ion beam in green wandering Jew (Tradescantia fluminensis)

• Altered levels of chlorophyll and anthocyanin contents, and abnormal ultrastructural features of chloroplasts were detected in the thermo-sensitive leaf-color mutant of green wandering Jew. • RNA-Seq analysis identified 42 candidate genes involved in photosynthesis, 14 candidate genes for chlorophyll biosynthesis, and 13 candidate genes for flavonoid and anthocyanin biosynthetic pathways. • Chlorophyll fluorescence signals, and HPLC confirmed the gene expression levels obtained by RNA-Seq. Leaf color is considered a valuable trait when breeding ornamental plants. In this study, a novel thermo-sensitive leaf-color mutant ( mt ) of green wandering Jew was obtained through carbon-ion beam irradiation. Young leaves of the mt plant exhibited pink coloration under low temperature ( mt _7 ℃) and green coloration under normal temperature ( mt _25 ℃). However, young leaves of the wild type (WT) remained green under low and normal temperatures. Here, we compared the mt and WT in terms of their anatomical, physiological, transcriptomic, and metabolomic characteristics at 7 ℃ and 25 ℃. Histological analyses showed that the upper epidermis and palisade parenchyma of mt were fewer and smaller. Chloroplasts of mt _25 ℃ were abnormally inflated, and leucoplasts were present instead of chloroplasts in mt _7 ℃ leaves. Physiological experiments also showed that mt _7 ℃ had lower chlorophyll and carotenoid contents, and higher anthocyanin contents; however, the chlorophyll content in mt _25 ℃ leaves were higher than that in the WT. Transcriptome profiling revealed that 99 differentially expressed genes (DEGs) and 1597 transcription factors (TFs) are involved in photosynthesis, chlorophyll biosynthesis, and flavonoid and anthocyanin biosyntheses. This study identified 8 candidate genes associated with the PS Ⅱ reaction center, 13 genes related to PS Ⅰ, 15 genes for LHC, 4 genes for PET/Cyt b6/f, and 2 genes for ATP synthase; these genes were differentially expressed at 7 ℃. DEGs related to photosynthesis and chlorophyll biosynthesis were down-regulated in the pink leaves of mt _7 ℃, whereas DEGs involved in flavonoid and anthocyanin biosyntheses were up-regulated in mt _7 ℃ and mt _25 ℃ leaves. Differential expression levels were further confirmed by qRT-PCR, Chlorophyll fluorescence signals, and previous HPLC results the. The anthocyanin content of mt _7 ℃ was 20-fold higher than that of the other pigments resulting in pink coloration at low temperature. This study provides a systematic evaluation of thermo-sensitive color variation in the mt . Based on these results, we identified key pathways related to thermo-sensitive leaf-color change in green wandering Jew.

A valine residue deletion in ZmSig2A, a sigma factor, accounts for a revertible leaf-color mutation in maize

A nuclear-encoded sigma (σ) factor is essential for the transcriptional regulation of plant chloroplast-encoded genes. Five putative maize σ factors have been identified by database searches, but their functions are unknown. We report a maize leaf color mutant etiolated/albino leaf 1 (eal1) that was derived from space mutation breeding. The eal1 mutant displays etiolated or albino leaves that then gradually turn to normal green at the seedling stage. The changes in eal1 leaf color are associated with changes in photosynthetic pigment content and chloroplast development. Map-based cloning revealed that a single amino-acid deletion changing Val480-Val481-Val482 to Val480-Val481, in the C-terminal domain σ4 of the putative σ factor ZmSig2A, is responsible for the eal1 mutation. In comparison with the expression level of the wild-type (WT) allele ZmSig2A+ in WT plants, much higher expression of the mutant allele ZmSig2AΔV in eal1 plants was detected before the eal1 plants turned to normal green. ZmSig2A shows the highest similarity to rice OsSig2A and Arabidopsis SIG2. Ectopic expression of ZmSig2A+ or ZmSig2AΔV driven by the cauliflower mosaic virus 35S promoter rescued the pale green leaf of the sig2 mutant, but ectopic expression of ZmSig2AΔV driven by the SIG2 promoter did not. We propose that the Val deletion generated a new weak allele of ZmSig2A that cannot completely abolish the ZmSig2A function. Some genes involved in chloroplast development and photosynthesis-associated nuclear genes showed significant expression differences between eal1 and WT plants. We conclude that ZmSig2A encoding a σ factor is essential for maize chloroplast development. The eal1 mutant with a weak allele of ZmSig2A represents a valuable genetic resource for investigating the regulation of ZmSig2A-mediated chloroplast development in maize. The eal1 mutation may be useful as a marker for early identification and elimination of false hybrids or transgene transmission in the application of genetic male sterility to commercial hybrid seed production.

Identification and fine mapping of a recessive gene controlling zebra leaf phenotype in maize.

The online version contains supplementary material available at 10.1007/s11032-021-01202-7.

A Mutation in CsYL2.1 Encoding a Plastid Isoform of Triose Phosphate Isomerase Leads to Yellow Leaf 2.1 (yl2.1) in Cucumber (Cucumis Sativus L.).

The leaf is an important photosynthetic organ and plays an essential role in the growth and development of plants. Leaf color mutants are ideal materials for studying chlorophyll metabolism, chloroplast development, and photosynthesis. In this study, we identified an EMS-induced mutant, yl2.1, which exhibited yellow cotyledons and true leaves that did not turn green with leaf growth. The yl2.1 locus was controlled by a recessive nuclear gene. The CsYL2.1 was mapped to a 166.7-kb genomic region on chromosome 2, which contains 24 predicted genes. Only one non-synonymous single nucleotide polymorphism (SNP) was found between yl2.1 and wt-WD1 that was located in Exon 7 of Csa2G263900, resulting in an amino acid substitution. CsYL2.1 encodes a plastid isoform of triose phosphate isomerase (pdTPI), which catalyzes the reversible conversion of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (GAP) in chloroplasts. CsYL2.1 was highly expressed in the cotyledons and leaves. The mesophyll cells of the yl2.1 leaves contained reduced chlorophyll and abnormal chloroplasts. Correspondingly, the photosynthetic efficiency of the yl2.1 leaves was impaired. Identification of CsYL2.1 is helpful in elucidating the function of ptTPI in the chlorophyll metabolism and chloroplast development and understanding the molecular mechanism of this leaf color variant in cucumber.

Differences in leaf anatomy, photosynthesis, and photoprotective strategies in the yellow-green leaf mutant and wild type of Rosa beggeriana Schrenk

Although the underlying mechanisms of chlorophyll biosynthesis have been intensively deciphered in model plants, leaf color mutants have not yet been fully studied in woody species. In this study, leaf anatomy, diurnal changes in photosynthesis and photoprotection were investigated in the yellow-green leaf (ygl) mutant of Rosa beggeriana. Chloroplast ultrastructure, pigment biosynthesis, and photosynthesis were impaired and led to photoinhibition at noon in the ygl mutant. The negative changes in leaf anatomy increased the risk of excess light absorption in the mutant. Moreover, the ygl mutant preferred to consume the excited energy through photorespiration rather than via heat dissipation. Finally, antioxidant defenses failed to scavenge reactive oxygen species and led to severe lipid peroxidation in the mutant. The results suggested that inhibition of photosynthesis in the ygl mutant of R. beggeriana was associated with the altered photoprotective strategies that involved in leaf anatomy, photorespiration, thermal dissipation, and antioxidant system.

GraS is critical for chloroplast development and affects yield in rice

Leaf color has been considered an important agronomic trait in rice (Oryza sativa L.) for a long time. The changes in leaf color affect the yield of rice. In this study, a green-revertible albino (graS) mutant was isolated from a 60Co-gamma-irradiated mutant pool of indica cultivar Guangzhan 63-4S. The fine mapping indicated that graS mutant was mapped to chromosome 1, and was located in a confined region between markers ab134 and InDel 8 with genetic distances of 0.11 and 0.06 cM, respectively. Based on the annotation results, four open reading frames (ORFs) were predicted in this region. Sequence analysis revealed that LOC_Os01g55974 had a 2-bp nucleotide insertion (AA) in the coding region that led to premature termination at the 324th base. Sequence analysis and expression analysis of related genes indicated that LOC_Os01g55974 is the candidate gene of GraS. We studied the genome and protein sequences of LOC_Os01g55974, and the data showed that GraS contains a deoxycytidine deaminase domain, which was expressed ubiquitously in all tissues. Further investigation indicated that GraS plays an essential role in the regulation of chloroplast biosynthesis, photosynthetic capacity and yield. Moreover, leaf color mutant can be used as an effective marker for the purity of breeding and hybridization.

Fine Mapping and Transcriptome Analysis of Virescent Leaf Gene v-2 in Cucumber (Cucumis sativus L.).

Leaf color mutants are the ideal materials to explore the pathways of chlorophyll metabolism, chloroplast development and photosynthesis system. In this study, a new virescent leaf mutant 104Y was identified by spontaneous mutation, whose cotyledon and upper five true leaves were yellow color. The yellow true leaves gradually turned green from top to bottom with increased chlorophyll contents. Genetic analysis indicated that the virescent leaf was controlled by one single recessive gene v-2, which was accurately mapped into 36.0–39.7 Mb interval on chromosome 3 by using BSA-seq and linkage analysis. Fine mapping analysis further narrowed v-2 into 73-kb genomic region including eight genes with BC1 and F2 populations. Through BSA-seq and cDNA sequencing analysis, only one nonsynonymous mutation existed in the Csa3G890020 gene encoding auxin F-box protein was identified, which was predicted as the candidate gene controlling virescent leaf. Comparative transcriptome analysis and quantitative real-time PCR analysis revealed that the expression level of Csa3G890020 was not changed between EC1 and 104Y. However, RNA-seq analysis identified that the key genes involved in chlorophyll biosynthesis and auxin signaling transduction network were mainly down-regulated in 104Y compared with EC1, which indicated that the regulatory functions of Csa3G890020 could be performed at post-transcriptional level rather than transcriptional level. This is the first report to map-based clone an auxin F-box protein gene related to virescent leaf in cucumber. The results will exhibit a new insight into the chlorophyll biosynthesis regulated by auxin signaling transduction network.

Mutation Mechanism of Leaf Color in Plants: A Review

Color mutation is a common, easily identifiable phenomenon in higher plants. Color mutations usually affect the photosynthetic efficiency of plants, resulting in poor growth and economic losses. Therefore, leaf color mutants have been unwittingly eliminated in recent years. Recently, however, with the development of society, the application of leaf color mutants has become increasingly widespread. Leaf color mutants are ideal materials for studying pigment metabolism, chloroplast development and differentiation, photosynthesis and other pathways that could also provide important information for improving varietal selection. In this review, we summarize the research on leaf color mutants, such as the functions and mechanisms of leaf color mutant-related genes, which affect chlorophyll synthesis, chlorophyll degradation, chloroplast development and anthocyanin metabolism. We also summarize two common methods for mapping and cloning related leaf color mutation genes using Map-based cloning and RNA-seq, and we discuss the existing problems and propose future research directions for leaf color mutants, which provide a reference for the study and application of leaf color mutants in the future.

Characterization and fine mapping of a leaf yellowing mutant in common wheat

Creation of leaf color mutants is the options in the study of photosynthesis, chloroplast development and Chlorophyll biosynthesis. In this study, a leaf color mutant NAU31 was identified from tissue culture of common wheat. In general, the mutant exhibited whole plant yellowing, fewer tillering, dwarfing and interestingly, improved resistance to wheat powdery mildew. Photosynthetic physiological analysis showed that the chloroplast inner structure varied in the mesophyll cells of the mutant, and this may lead to reduced chlorophyll content (chlorophyll “a”, Chlorophyll “b”, total chlorophyll and carotenoid), as well as decreased maximum quantum efficiency of photosystem II (Fv/Fm) and net photosynthetic rate. Transcriptome analysis indicated that the impairing of photosynthesis may be responsible for plant yellowing of NAU31. Inheritance analysis showed that NAU31 was a gain-of-function mutant and the yellowing trait was controlled by a dominant gene, namely Taymg (yellow mutant gene). SSR markers analysis preliminarily mapped Taymg to the distal short arm of chromosome 2D using an F2 population from the cross between NAU31 and W7984, a synthesized wheat. The mapping result was confirmed by BSA using a 55 K wheat SNP array. Through recombinants screening, the Taymg was finely mapped to the terminal region of 2DS, flanked by markers ZH47 and gwm296 and corresponding to a physical interval of 0.76 Mb. The cloning of Taymg will facilitate the dissection of the mechanism involved in chloroplast development of wheat and the coordinating of plant development and disease resistance.

Transcriptome analysis reveals genes associated with leaf color mutants in Cymbidium longibracteatum

Cymbidium longibracteatum is an economically important Chinese orchid well known for its silver- or gold-colored leaf color patterns, and is desired for its ornamental value. In this study, we analyzed the yellow and green rhizomes from C. longibracteatum by deep RNA-sequencing. A total of 333,497 transcripts and 162,407 unigenes of ‘longchangsu’ obtained from 39.59 Gb of clean reads. Differentially expressed gene (DEG) analysis indicated that 497 genes were upregulated and 147 downregulated in the leaf color mutant. Five of these DEGs were involved in the phototransduction pathway, and 48 genes belonged to the plant-pathogen interaction pathway. Furthermore, we identified seven genes associated with the photosystem, six of which were upregulated, and one (psbA) was downregulated. Our results showed that downregulation of psbA gene may produce leaf color mutants under certain culture conditions that affect the D1 protein of photosystem II. All seven genes were validated by qRT-PCR. This study on genes related to photosystem and other pathways establishes a genetic foundation for ornamental orchids and may help to improve the economic and breeding values of C. longibracteatum.

Double mutation of two homologous genes YL1 and YL2 results in a leaf yellowing phenotype in soybean [Glycine max (L.) Merr

Two homologous, chloroplast located CAAX proteases were identified to be functional redundancy in determining soybean leaf color, and they probably play essential roles in regulating light harvesting and absorption during photosynthesis process. Leaf color mutants are ideal materials for studying photosynthesis and chlorophyll metabolism. The soybean [Glycine max (L.) Merr.] yellowing leaf (yl) variation is a recombinant mutant characterized by yellow foliage, which derived from the specific cross between green seed-coated and yellow seed-coated soybean varieties. Molecular cloning and subsequent gene silencing revealed that the yellow leaf trait of yl was controlled by two recessive nuclear genes, glyma11g04660 and glyma01g40650, named as YL1 and YL2 respectively, and the latter was confirmed to be same as the earlier reported green seed-coat gene G. Both YL1 and YL2 belonged to chloroplast-located proteases possessing Abi domain, and these genes were expressed in various tissues, especially in young leaves. In yl, the expression of YL1 and YL2 were suppressed in most tissues, and the young leaf of yl presented an increased maximal photochemical efficiency (Fv/Fm) as well as enhanced net photosynthesis activity (Pn), indicating that YL1 and YL2 are involved in light absorption regulation during photosynthesis process. Collectively, the identification and description of YL1 and YL2 in our study provides insights for the regulatory mechanism of photosynthesis process, and these findings will further assist to clarify the close relationship between photosynthesis and chlorophyll metabolism.

Dark/Light Treatments Followed by γ-Irradiation Increase the Frequency of Leaf-Color Mutants in Cymbidium.

Radiation randomly induces chromosomal mutations in plants. However, it was recently found that the frequency of flower-color mutants could be specifically increased by upregulating anthocyanin pathway gene expression before radiation treatments. The mechanisms of chlorophyll biosynthesis and degradation are active areas of plant study because chlorophyll metabolism is closely connected to photosynthesis. In this study, we determined the dark/light treatment conditions that resulted in upregulation of the expression levels of six chlorophyll pathway genes, uroporphyrinogen III synthase (HEMD), uroporphyrinogen III decarboxylase (HEME2), NADPH-protochlorophyllide oxidoreductase (POR) A (PORA), chlorophyll synthase (CHLG), chlorophyllase (CLH2), and red chlorophyll catabolite reductase (RCCR), and measured their effects on the γ-irradiation-induced frequencies of leaf-color mutants in two Cymbidium cultivars. To degrade chlorophyll in rhizomes, 60–75 days of dark treatment were required. To upregulate the expressions of chlorophyll pathway genes, 10 days of light treatment appeared to be optimal. Dark/light treatments followed by γ-irradiation increased chlorophyll-related leaf mutants by 1.4- to 2.0-fold compared with γ-ray treatment alone. Dark/light treatments combined with γ-irradiation increased the frequency of leaf-color mutants in Cymbidium, which supports the wider implementation of a plant breeding methodology that increases the mutation frequency of a target trait by controlling the expression of target trait-related genes.

The identification of key candidate genes mediating yellow seedling lethality in a Lilium regale mutant.

Leaf color mutants are ideal materials for exploring plant photosynthesis mechanisms, chlorophyll biosynthetic pathways and chloroplast development. The yellow seedling lethal mutant lrysl1 was discovered from self-bred progenies of Lilium regale; however, the mechanism of leaf color mutation remains unclear. In this study, the ultrastructural and physiological features and de novo RNA-Seq data of a L. regale leaf color mutant and wild-type L. regale were investigated. Genetic analysis indicated that the characteristics of the lrysl1 mutant were controlled by a recessive nuclear gene. The chlorophyll a, chlorophyll b and carotenoid contents in the mutant leaves were lower than those in the wild-type leaves. Furthermore, the contents of the chlorophyll precursors aminolevulinic acid (ALA), porphobilinogen (PBG), protoporphyrin IX (ProtoIX), Mg-protoporphyrin IX (Mg-ProtoIX), and protochlorophyll (Pchl) decreased significantly in mutant leaves. Transcriptome data from the mutant and wild type showed that a total of 892 differentially expressed genes were obtained, of which 668 and 224 were upregulated genes and downregulated genes in the mutant, respectively. Almost all genes in the photosynthesis pathway and chlorophyll biosynthetic pathway were downregulated in the mutant, which corroborated the differences in the physiological features mentioned above. Further research indicated that the chloroplasts of the mutant leaves exhibited an abnormal morphology and distribution and that the expression of a gene related to chloroplast development was downregulated. It was concluded that abnormal chloroplast development was the main cause of leaf color mutation in the mutant lrysl1 and that LrGLK was a gene related to chloroplast development in L. regale. This research provides a foundation for further research on the mechanism by which LrGLK regulates chloroplast development in L. regale.

Identification of two recessive etiolation genes (py1, py2) in pakchoi (Brassica rapa L. ssp. chinensis)

BackgroundLeaf color is a major agronomic trait, which has a strong influence on crop yields. Isolating leaf color mutants can represent valuable materials for research in chlorophyll (Chl) biosynthesis and metabolism regulation.ResultsIn this study, we identified a stably inherited yellow leaf mutant derived from ‘Huaguan’ pakchoi variety via isolated microspore culture and designated as pylm. This mutant displayed yellow leaves after germination. Its etiolated phenotype was nonlethal and stable during the whole growth period. Its growth was weak and its hypocotyls were markedly elongated. Genetic analysis revealed that two recessive nuclear genes, named py1 and py2, are responsible for the etiolation phenotype. Bulked segregant RNA sequencing (BSR-Seq) showed that py1 and py2 were mapped on chromosomes A09 and A07, respectively. The genes were single Mendelian factors in F3:4 populations based on a 3:1 phenotypic segregation ratio. The py1 was localized to a 258.3-kb interval on a 34-gene genome. The differentially expressed gene BraA09004189 was detected in the py1 mapping region and regulated heme catabolism. One single-nucleotide polymorphism (SNP) of BraA09004189 occurred in pylm. A candidate gene-specific SNP marker in 1520 F3:4 yellow-colored individuals co-segregated with py1. For py2, 1860 recessive homozygous F3:4 individuals were investigated and localized py2 to a 4.4-kb interval. Of the five genes in this region, BraA07001774 was predicted as a candidate for py2. It encoded an embryo defective 1187 and a phosphotransferase related to chlorophyll deficiency and hypocotyl elongation. One SNP of BraA07001774 occurred in pylm. It caused a single amino acid mutation from Asp to Asn. According to quantitative real-time polymerase chain reaction (qRT-PCR), BraA07001774 was downregulated in pylm.ConclusionsOur study identified a Chl deficiency mutant pylm in pakchoi. Two recessive nuclear genes named py1 and py2 had a significant effect on etiolation. Candidate genes regulating etiolation were identified as BraA09004189 and BraA07001774, respectively. These findings will elucidate chlorophyll metabolism and the molecular mechanisms of the gene interactions controlling pakchoi etiolation.