<Emphasis Type="Italic">Bacillus subtilis</Emphasis> based-formulation for the control of postbloom fruit drop of citrus

Postbloom fruit drop (PFD) caused by Colletotrichum acutatum affects flowers and causes early fruit drop in all commercial varieties of citrus. Biological control with the isolate ACB-69 of Bacillus subtilis has been considered as a potential method for controlling this disease. This study aimed to develop and optimize a B. subtilis based-formulation with a potential for large-scale applications and evaluate its effect on C. acutatum in vitro and in vivo. Bacillus subtilis based-formulations were developed using different carrier materials, and their ability to control PFD was evaluated. The results of the assays led to the selection of the B. subtilis based-formulation with talc + urea (0.02 %) and talc + ammonium molybdate (1 mM), which inhibited mycelial growth and germination of C. acutatum. Studies with detached citrus flowers showed that the formulations were effective in controlling the pathogen. In field conditions, talc + urea (0.02 %) provided 73 % asymptomatic citrus flowers and 56 % of the average number of effective fruit (ANEF), equating with fungicide treatment. On the contrary, non-treated trees had 8.8 % of asymptomatic citrus flowers and 0.83 % ANEF. The results suggest that B. subtilis based-formulations with talc as the carrier supplemented with a nitrogen source had a high potential for PFD control.     

Extrafloral nectaries of four varieties of <Emphasis Type="Italic">Chamaecrista ramosa</Emphasis> (Vogel) H.S.Irwin &amp; Barneby (Fabaceae): anatomy,chemical nature,mechanisms of nectar secretion,and elimination

Considering the importance of extrafloral nectaries (EFNs) in Fabaceae, the objectives of this research were to analyze (1) the anatomical and histochemical characteristics of the EFNs of Chamaecrista ramosa var. ramosa, C. ramosa var. curvifoliola, C. ramosa var. parvifoliola, and C. ramosa var. lucida and (2) the ultrastructure of the EFNs of C. ramosa var. ramosa. Standard techniques in plant anatomy and transmission electron microscopy were used. The anatomical analyses confirmed the characteristics described for extrafloral nectaries, evidencing three well-defined regions: epidermis, nectariferous, and subnectariferous parenchymas. Carbohydrates, proteins, pectins/mucilages, and lipids were detected by histochemical analyzes in all varieties. The ultrastructure of the EFNs of C. ramosa var. ramosa allowed the observation of microchannels at the external periclinal cell walls of the epidermis covering the secretory region. The nectariferous and subnectariferous parenchyma cells have periplasmic spaces, large plastids containing starch grains and plastoglobules, mitochondria, developed endoplasmic reticulum, large vacuoles with electron-dense contents, and membrane residues may be associated with the vacuole, suggesting the occurrence of autophagic processes. The anatomical, histochemical, and ultrastructural patterns revealed characteristics that confirm the glands of C. ramosa as extrafloral nectaries and suggest the eccrine mechanism of secretion.     

Fine structure of the <Emphasis Type="Italic">Arabidopsis</Emphasis> stem cuticle: effects of fixation and changes over development

Main conclusion

The Arabidopsis cuticle, as observed by electron microscopy, consists primarily of the cutin/cutan matrix. The cuticle possesses a complex substructure, which is correlated with the presence of intracuticular waxes. The plant cuticle is composed of an insoluble polyester, cutin, and organic solvent soluble cuticular waxes, which are embedded within and coat the surface of the cutin matrix. How these components are arranged in the cuticle is not well understood. The Arabidopsis cuticle is commonly understood as ‘amorphous,’ lacking in ultrastructural features, and is often observed as a thin (~80–100 nm) electron-dense layer on the surface of the cell wall. To examine this cuticle in more detail, we examined cuticles from both rapidly elongating and mature sections of the stem and compared the preservation of the cuticles using conventional chemical fixation methods and high-pressure freezing/freeze-substitution (HPF/FS). We found that HPF/FS preparation revealed a complex cuticle substructure, which was more evident in older stems. We also found that the cuticle increases in thickness with development, indicating an accretion of polymeric material, likely in the form of the non-hydrolyzable polymer, cutan. When wax was extracted by chloroform immersion prior to sample preparation, the contribution of waxes to cuticle morphology was revealed. Overall, the electron-dense cuticle layer was still visible but there was loss of the cuticle substructure. Furthermore, the cuticle of cer6, a wax-deficient mutant, also lacked this substructure, suggesting that these fine striations were dependent on the presence of cuticular waxes. Our findings show that HPF/FS preparation can better preserve plant cuticles, but also provide new insights into the fine structure of the Arabidopsis cuticle.

     

Overexpression of the ABC transporter gene <Emphasis Type="Italic">TsABCG11</Emphasis> increases cuticle lipids and abiotic stress tolerance in <Emphasis Type="Italic">Arabidopsis</Emphasis>

The cuticle, composed primarily of wax and cutin, covers most plant aerial surfaces and plays a vital role in interactions between plants and their environment. Some ATP-binding cassette G subfamily (ABCG) members are involved in cuticular lipid molecule exportation to outside in the plant surface. Thellungiella salsugineum, a relative of Arabidopsis thaliana with a heavy cuticle, has extreme stress tolerance. TsABCG11, an ABCG transporter was cloned (GenBank accession number JQ389853), and its structure was studied. qRT-PCR showed that TsABCG11 expression varied in different organs of T. salsugineum and was upregulated under ABA, NaCl, drought and cold conditions. The rosette leaves from 4-week-old TsABCG11 overexpressed (OE) Arabidopsis plants displayed lower rates of water loss and decreased chlorophyll-extracted rates compared to wild-type plants. TsABCG11-OE plants also exhibited significantly increased total cuticular wax and cutin monomer amounts, mainly due to prominent changes in the C₂₉, C₃₁, and C₃₃ alkanes in the wax and C18:2 dioic in cutin monomers, respectively. TsABCG11-OE seedlings exhibit lower root growth inhibition under 100 mM of NaCl or 1 µM of ABA than the wild type. Four-week-old TsABCG11-OE plants exhibited higher photosynthetic rates and water-use efficiency under cold stress (4 °C) than control plants. These results indicate that TsABCG11 plays an important role in cuticle lipid exportation and is involved in abiotic stresses, probably having a close relationship with extreme stress tolerance in T. salsugineum.     

Visualization of Endoplasmic Reticulum and Mitochondria in <Emphasis Type="Italic">Aurantiochytrium limacinum</Emphasis> by the Expression of EGFP with Cell Organelle-Specific Targeting/Retaining Signals

Thraustochytrids are single cell marine eukaryotes that produce large amounts of polyunsaturated fatty acids such as docosahexaenoic acid. In the present study, we report the visualization of endoplasmic reticulum (ER) and mitochondria in a type strain of the thraustochytrid, Aurantiochytrium limacinum ATCC MYA-1381, using the enhanced green fluorescent protein (EGFP) with specific targeting/retaining signals. We expressed the egfp gene with ER targeting/retaining signals from A. limacinum calreticulin or BiP/GRP78 in the thraustochytrid, resulting in the distribution of EGFP signals at the perinuclear region and near lipid droplets. ER-Tracker? Red, an authentic fluorescent probe for the visualization of ER in mammalian cells, also stained the same region. We observed small lipid droplets generated from the visualized ER in the early growth phase of cell culture. Expression of the egfp gene with the mitochondria targeting signal from A. limacinum cytochrome c oxidase resulted in the localization of EGFP near the plasma membrane. The distribution of EGFP signals coincided with that of MitoTracker® Red CMXRos, which is used to visualize mitochondria in eukaryotes. The ER and mitochondria of A. limacinum were visualized for the first time by EGFP with thraustochytrid cell organelle-specific targeting/retaining signals. These results will contribute to classification of the intracellular localization of proteins expressed in ER and mitochondria as well as analyses of these cell organelles in thraustochytrids.     

Endoplasmic reticulum stress and calcium imbalance are involved in cadmium-induced lipid aberrancy in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The endoplasmic reticulum is the key organelle which controls protein folding, lipid biogenesis, and calcium (Ca²⁺) homeostasis. Cd exposure in Saccharomyces cerevisiae activated the unfolded protein response and was confirmed by the increased Kar2p expression. Cd exposure in wild-type (WT) cells increased PC levels and the PC biosynthetic genes. Deletion of the two phospholipid methyltransferases CHO2 and OPI3 modulated PC, TAG levels and the lipid droplets with cadmium exposure. Interestingly, we noticed an increase in the calcium levels upon Cd exposure in the mutant cells. This study concluded that Cd interrupted calcium homeostasis-induced lipid dysregulation leading to ER stress.     

Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. isolated from chili pepper fruit exhibiting symptoms of anthracnose in Thailand

Colletotrichum spp. are causal agents of anthracnose disease in chili fruits and other tropical crops. The disease is increasing in chili fruits in Thailand and significantly reduces fruit quality and fruit production. Forty-eight isolates of Colletotrichum spp. associated with chili anthracnose were collected from different areas of Thailand during 2010–2015. Based on morphological characteristic identification, 10 isolates were shown to belong to the C. gloeosporioides species complex, 24 isolates belong to the C. acutatum species complex and 14 isolates to C. capsici. For molecular identification, two primer sets, ITS1/ITS4 and ACT528/ACT738, were used for amplification of the internal transcribed spacer of rRNA gene (ITS1–5.8S–ITS2) and partial region actin gene (ACT), respectively. The phylogenetic analysis of individual and combined ITS region and actin nucleotide sequences identified the collected isolates into 4 species: C. gloeosporioides, C. siamense, C. acutatum and C. capsici. The pathogenicity test demonstrated that all four species were pathogenic on intact unwounded and healthy fruits. These results indicated that C. capsici, C. acutatum, C. gloeosporioides and C. siamense were the causal agents of chili anthracnose disease.     

Structure of the stigma and style of <Emphasis Type="Italic">Callaeum psilophyllum</Emphasis> (Malpighiaceae) and its relation with potential pollinators

The family Malpighiaceae, particularly in the Neotropic, shows a similar floral morphology. Although floral attraction and rewards to pollinators are alike, stigmas and styles show more diversity. The stigmas were described covered with a thin and impermeable cuticle that needs to be ruptured by the mechanical action of the pollinators. However, this characteristic was only mentioned for a few species and the anatomy and ultrastructure of the stigmas were not explored. In this work, we analyze the morphology, anatomy, and ultrastructure of the stigma and style of Callaeum psilophyllum. Moreover, we identify the potential pollinators in order to evaluate how the disposition of the stigmas is related with their size and its role in the exposure of the receptive stigmatic surface. Our observations indicate that Centris flavifrons, C. fuscata, C. tarsata, and C. trigonoides are probably efficient pollinators of C. psilophyllum. The three stigmas are covered by a cuticle that remained intact in bagged flowers. The flowers exposed to visitors show the cuticle broken, more secretion in the intercellular spaces between sub-stigmatic cells and abundant electron-dense components inside vacuoles in stigmatic papillae. This indicates that the stigmas prepares in similar ways to receive pollen grains, but the pollinator action is required to break the cuticle, and once pollen tubes start growing, stigmatic and sub-stigmatic cells release more secretion by a granulocrine process.     

Cloning of the <Emphasis Type="Italic">Brcer1</Emphasis> gene involved in cuticular wax production in a glossy mutant of non-heading Chinese cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L. var. <Emphasis Type="Italic">communis</Emphasis>)

Cuticular wax is a complex mixture of very-long-chain fatty acid derivatives. The wax on the surface of plants serves as a protective barrier to reduce non-stomatal water loss and environmental damage. However, the loss of wax may lead to a glossy phenotype, which is an favorable trait in leafy vegetables. The mechanism of glossy mutants in non-heading Chinese cabbage (Brassica rapa L. var. communis) has not been studied yet. In this study, scanning electron microscopy (SEM) showed that the cuticular wax on the leaves and stem of a glossy mutant was dramatically reduced compared with that of the wild-type plant. Transmission electron microscopy (TEM) revealed that the cuticle ultrastructure of glossy mutant leaf and stem were altered when compared with the wild type. A cuticle wax analysis showed the total wax content of leaves, as well as alkanes, ketones and alcohols, was decreased. A genetic analysis indicated that the glossy phenotype was controlled by a single gene. Based on a homology analysis, the Brcer1 gene was identified as the candidate gene controlling the glossy phenotype. In the glossy mutant, a 39-bp deletion leads to an mRNA disruption and reduces the expression of the BrCER1 gene. Sequence analysis showed that a loss of function mutation in the Brcer1 gene was different from that of Cgl1, which was previously shown to be responsible for the glossy phenotype in B. oleracea, showing typical parallel selection. These findings provide a better understanding of the cuticular wax biosynthesis pathway and offer important information for molecular-assisted breeding of non-heading Chinese cabbage (B. rapa L. var. communis).     

The expression level of anthocyanidin synthase determines the anthocyanin content of crabapple (<Emphasis Type="Italic">Malus</Emphasis> sp.) petals

In addition to contributing to the coloration of plant organs and their defense against herbivores, the consumption of anthocyanins in the human diet has a number of health benefits. Crabapple (Malus sp.) represents a valuable experimental model system to research the mechanisms and regulation of anthocyanin accumulation, in part due to the often vivid and varied petal and leaf coloration that is exhibited by various cultivars. The enzyme anthocyanidin synthase (ANS) plays a pivotal role in anthocyanin biosynthesis; however, the relationship between ANS expression and petal pigmentation has yet to be established in crabapple. To illuminate the mechanism of anthocyanin accumulation in crabapple petals, we evaluated the expression of two crabapple ANS allelic genes (McANS-1 and McANS-2) and the levels of anthocyanins in petals from cultivars with dark red (‘Royalty’) and white (‘Flame’) petals, as well as another (‘Radiant’) whose petals have an intermediate pink color. We determined that the expression of McANS in the three cultivars correlated with the variation of anthocyanin accumulation during different petal developmental stages. Furthermore, transgenic tobacco plants constitutively overexpressing one of the two McANS genes, McANS-1, had showed elevated anthocyanin accumulation and a deeper red coloration in their petals than those from untransformed control lines. In conclusion, we propose that McANS are responsible for anthocyanin accumulation during petal coloration in different crabapple cultivars.     

Alteration of flower colour in <Emphasis Type="Italic">Ipomoea nil</Emphasis> through CRISPR/Cas9-mediated mutagenesis of <Emphasis Type="Italic">carotenoid cleavage dioxygenase 4</Emphasis>

Japanese morning glory, Ipomoea nil, exhibits a variety of flower colours, except yellow, reflecting the accumulation of only trace amounts of carotenoids in the petals. In a previous study, we attributed this effect to the low expression levels of carotenogenic genes in the petals, but there may be other contributing factors. In the present study, we investigated the possible involvement of carotenoid cleavage dioxygenase (CCD), which cleaves specific double bonds of the polyene chains of carotenoids, in the regulation of carotenoid accumulation in the petals of I. nil. Using bioinformatics analysis, seven InCCD genes were identified in the I. nil genome. Sequencing and expression analyses indicated potential involvement of InCCD4 in carotenoid degradation in the petals. Successful knockout of InCCD4 using the CRISPR/Cas9 system in the white-flowered cultivar I. nil cv. AK77 caused the white petals to turn pale yellow. The total amount of carotenoids in the petals of ccd4 plants was increased 20-fold relative to non-transgenic plants. This result indicates that in the petals of I. nil, not only low carotenogenic gene expression but also carotenoid degradation leads to extremely low levels of carotenoids.     

Morphology,ultrastructure, and chemical compounds of the osmeterium of <Emphasis Type="Italic">Heraclides thoas</Emphasis> (Lepidoptera: Papilionidae)

The osmeterium, found in papilionoid larvae, is an eversible organ with an exocrine gland that produces substances in response to the mechanical disturbances caused by natural enemies. The anatomy, histology and ultrastructure of the osmeterium, and the chemical composition of its secretion in Heraclides thoas (Lepidoptera: Papilionidae) were studied. Heraclides thoas larvae have a Y-shaped osmeterium in the thorax. The surface of the osmeterium has a rough cuticle lining cells with papillae and irregular folds, whereas the cells that limited the gland pores are irregular, folded, and devoid of papillae. Two types of cells are found: (i) cuticular epidermal cells on the surface of the tubular arms of the osmeterium and (ii) secretory cells of the ellipsoid gland within the region of the glandular pore. Cuticular epidermal cells show a thick cuticle, with several layers divided into epicuticle and lamellar endocuticle. Secretory cells are polygonal, with extensive folds in the basal plasma membrane that formed extracellular channels. The cytoplasm has mitochondria, ribosomes, and numerous vacuoles, whereas the nucleus is irregular in shape with decondensed chromatin. The chemical composition of the osmeterial secretion comprised (Z)-α-bisabolene (25.4%), α-bisabol (20.6%), β-bisabolene (13.1%), (E)-α-bisabolene 8%), β-pinene (9.91%), longipinene epoxide (8.92%), (Z)-β-farnesene (6.96%), β-caryophyllene (2.05%), farnesol (1.86%), linalyl propionate (1.86%), and 1-octyn-4-ol (1.07%). The morphological features suggest that the cuticular epidermal cells play a major role in the maintenance and protection of the osmeterium, whereas secretory cells are responsible for production of osmeterial secretions.     

Monoterpene biotransformation by <Emphasis Type="Italic">Colletotrichum</Emphasis> species

Objective

To investigate the biocatalytic potential of Colletotrichum acutatum and Colletotrichum nymphaeae for monoterpene biotransformation.

Results

C. acutatum and C. nymphaeae used limonene, α-pinene, β-pinene, farnesene, citronellol, linalool, geraniol, perillyl alcohol, and carveol as sole carbon and energy sources. Both species biotransformed limonene and linalool, accumulating limonene-1,2-diol and linalool oxides, respectively. α-Pinene was only biotransformed by C. nymphaeae producing campholenic aldehyde, pinanone and verbenone. The biotransformation of limonene by C. nymphaeae yielded 3.34–4.01 g limonene-1,2-diol l^?1, depending on the substrate (R-(+)-limonene, S-(?)-limonene or citrus terpene (an agro-industrial by-product). This is among the highest concentrations already reported for this product.

Conclusions

This is the first report on the biotransformation of these terpenes by Colletotrichum spp. and the biotransformation of limonene to limonene-1,2-diol possibly involves enzymes similar to those found in Grosmannia clavigera.

     

Functional micromorphology of petals of <Emphasis Type="Italic">Chaenomeles japonica</Emphasis> exposed to humid and cold season

In this study, micro- and nano-traits of petal epidermises of flowers of Chaenomeles japonica extended under environmental conditions, during the humid and cold period of the year, are presented. The outer (abaxial) and the inner (adaxial) epidermises of petals of C. japonica consist of convex and papillae cells, respectively, that are covered by epicuticular wrinkled relief further ornamented by submicron motifs, forming interfaces between floral tissues and environment. Structural epidermal features of the petal relief at the nanoscale level reveal different functionality on the two sides of the corolla. The cuticular folds of convex epidermal cells display declining water retention on the outer petal surface and the exposed side of the corolla to the environmental conditions. The cuticular folds of papillae epidermal cells increase in size the inner petal surface, in comparison with the outer surface; such traits facilitate light absorption and enhanced the contact area among folds and curvatures at the inner side of the corolla. It appears that nanometric surface structures of petals may be important adaptive features of C. japonica flowers, contributing to their performance in the field.     

Metabolic alteration of <Emphasis Type="Italic">Catharanthus roseus</Emphasis> cell suspension cultures overexpressing <Emphasis Type="Italic">geraniol synthase</Emphasis> in the plastids or cytosol

Previous studies showed that geraniol could be an upstream limiting factor in the monoterpenoid pathway towards the production of terpenoid indole alkaloid (TIA) in Catharanthus roseus cells and hairy root cultures. This shortage in precursor availability could be due to (1) limited expression of the plastidial geraniol synthase resulted in a low activity of the enzyme to catalyze the conversion of geranyl diphosphate to geraniol; or (2) the limitation of geraniol transport from plastids to cytosol. Therefore, in this study, C. roseus’s geraniol synthase (CrGES) gene was overexpressed in either plastids or cytosol of a non-TIA producing C. roseus cell line. The expression of CrGES in the plastids or cytosol was confirmed and the constitutive transformation lines were successfully established. A targeted metabolite analysis using HPLC shows that the transformed cell lines did not produce TIA or iridoid precursors unless elicited with jasmonic acid, as their parent cell line. This indicates a requirement for expression of additional, inducible pathway genes to reach production of TIA in this cell line. Interestingly, further analysis using NMR-based metabolomics reveals that the overexpression of CrGES impacts primary metabolism differently if expressed in the plastids or cytosol. The levels of valine, leucine, and some metabolites derived from the shikimate pathway, i.e. phenylalanine and tyrosine were significantly higher in the plastidial- but lower in the cytosolic-CrGES overexpressing cell lines. This result shows that overexpression of CrGES in the plastids or cytosol caused alteration of primary metabolism that associated to the plant cell growth and development. A comprehensive omics analysis is necessary to reveal the full effect of metabolic engineering.     

Functional analysis of <Emphasis Type="Italic">PI-</Emphasis>like gene in relation to flower development from bamboo (<Emphasis Type="Italic">Bambusa oldhamii</Emphasis>)

Bamboo flowering owns many unique characteristics and remains a mystery. To investigate the molecular mechanisms underlying flower development in bamboo, a petal-identity gene was identified as a PISTILLATA homologue named BoPI from Bambusa oldhamii (bamboo family). Expression analysis showed that BoPI was highly expressed in flower organs and gradually increased during flower development stage, suggesting that BoPI played an important role in flower development. Ectopic expression of BoPI in Arabidopsis caused conversion of sepals to petals. 35S::BoPI fully rescued the defective petal formation in the pi-1 mutant. BoPI could interact with BoAP3 protein in vitro. These results suggested that BoPI regulated flower development of bamboo in a similar way with PI. Besides flower organs, BoPI was also expressed in leaf and branch, which revealed that BoPI may involve in leaf and branch development. Similar to other MIKC-type gene, BoPI contained the C-terminal sequence but its function was controversial. Ectopic expression of the C-terminal deletion construct (BoPI- ΔC) in Arabidopsis converted sepals to petals; BoPI- ΔC interacted with BoAP3 on yeast two-hybrid assay, just like the full-length construct. The result implied that the C-terminal sequence may not be absolutely required for organ identity function in the context of BoPI.     

Nuclear genome sequence of the plastid-lacking cryptomonad <Emphasis Type="Italic">Goniomonas avonlea</Emphasis> provides insights into the evolution of secondary plastids

Background

The evolution of photosynthesis has been a major driver in eukaryotic diversification. Eukaryotes have acquired plastids (chloroplasts) either directly via the engulfment and integration of a photosynthetic cyanobacterium (primary endosymbiosis) or indirectly by engulfing a photosynthetic eukaryote (secondary or tertiary endosymbiosis). The timing and frequency of secondary endosymbiosis during eukaryotic evolution is currently unclear but may be resolved in part by studying cryptomonads, a group of single-celled eukaryotes comprised of both photosynthetic and non-photosynthetic species. While cryptomonads such as Guillardia theta harbor a red algal-derived plastid of secondary endosymbiotic origin, members of the sister group Goniomonadea lack plastids. Here, we present the genome of Goniomonas avonlea—the first for any goniomonad—to address whether Goniomonadea are ancestrally non-photosynthetic or whether they lost a plastid secondarily.

Results

We sequenced the nuclear and mitochondrial genomes of Goniomonas avonlea and carried out a comparative analysis of Go. avonlea, Gu. theta, and other cryptomonads. The Go. avonlea genome assembly is ~?92 Mbp in size, with 33,470 predicted protein-coding genes. Interestingly, some metabolic pathways (e.g., fatty acid biosynthesis) predicted to occur in the plastid and periplastidal compartment of Gu. theta appear to operate in the cytoplasm of Go. avonlea, suggesting that metabolic redundancies were generated during the course of secondary plastid integration. Other cytosolic pathways found in Go. avonlea are not found in Gu. theta, suggesting secondary loss in Gu. theta and other plastid-bearing cryptomonads. Phylogenetic analyses revealed no evidence for algal endosymbiont-derived genes in the Go. avonlea genome. Phylogenomic analyses point to a specific relationship between Cryptista (to which cryptomonads belong) and Archaeplastida.

Conclusion

We found no convincing genomic or phylogenomic evidence that Go. avonlea evolved from a secondary red algal plastid-bearing ancestor, consistent with goniomonads being ancestrally non-photosynthetic eukaryotes. The Go. avonlea genome sheds light on the physiology of heterotrophic cryptomonads and serves as an important reference point for studying the metabolic “rewiring” that took place during secondary plastid integration in the ancestor of modern-day Cryptophyceae.

     

Fine mapping of <Emphasis Type="Italic">BoGL1</Emphasis>, a gene controlling the glossy green trait in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. Var. <Emphasis Type="Italic">capitata</Emphasis>)

The cuticular wax covering epidermal cells causes the glaucous appearance in cabbage. As a protective barrier, cuticular wax plays various roles in protection against biotic and abiotic stresses. This is the first gene mapping report of a dominant glossy green cabbage mutant. In the present paper, scanning electron microscopy (SEM) demonstrated that the wax crystals were severely reduced in the mutant, which indicates that the glossy green phenotype is caused by cuticular wax reduction. Genetic analysis revealed that the glossy trait is controlled by a single dominant gene. Through primer screening and fine mapping, the mutant gene BoGL1 (Brassica oleracea glossy 1) was delimited to the end of chromosome C08 by the flanking marker SSRC08–76 at a genetic distance of 0.2 cM. Two genes homologous to CER1 (ECERIFERUM 1), a gene related to wax biosynthesis in Arabidopsis, were located in the mapped region. Expressional analysis revealed that the Bol018504 gene was severely suppressed but that no nucleotide variation was found by sequencing. These results lay the foundation for the functional analysis of BoGL1, and they will accelerate the research on wax metabolism in cabbage.