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Stock (Matthiola incana (L.) R. Br.) is a famous annual ornamental plant with important ornamental and economic value. The lack of DNA molecular markers has limited genetic analysis, genome evolution, and marker-assisted selective breeding studies of M. incana. Therefore, more DNA markers are needed to support the further elucidation of the biology and genetics of M. incana. In this study, a high-quality genome of M. incana was initially assembled and a set of effective SSR primers was developed at the whole-genome level using genome data. A total of 45,612 loci of SSRs were identified; the di-nucleotide motifs were the most abundant (77.35%). In total, 43,540 primer pairs were designed, of which 300 were randomly selected for PCR validation, and as the success rate for amplification. In addition, 22 polymorphic SSR markers were used to analyze the genetic diversity of 40 stock varieties. Clustering analysis showed that all varieties could be divided into two clusters with a genetic distance of 0.68, which were highly consistent with their flower shape (potted or cut type). Moreover, we have verified that these SSR markers are effective and transferable within the Brassicaceae family. In this study, potential SSR molecular markers were successfully developed for 40 M. incana varieties using whole genome analysis, providing an important genetic tool for theoretical and applied research on M. incana.

Introducción y objetivos: El género Matthiola W. T Aiton posee entre 50 y 60 especies originarias del Mediterráneo, Asia central y Sudáfrica. M. incana (L.) W. T. Aiton, nativa del sur de Europa, se encuentra naturalizada en otras regiones del mundo. En Argentina es cultivada como ornamental. A partir de observaciones florísticas se halló una población naturalizada, que se autoperpetúa sin intervención del hombre, en las costas de la Patagonia Argentina. El objetivo de este trabajo es reportar por primera vez la naturalización de M. incana en Argentina. M&M: Para la identificación de la especie se siguieron los métodos clásicos de taxonomía, se consultó bibliografía específica y se contrastó el material recolectado con imágenes digitales del lectotipo. Se recolectaron semillas de la población naturalizada y se evaluó el poder germinativo a través de un ensayo de germinación. El material recolectado que documenta el presente trabajo fue depositado en el herbario ARC de la Universidad Nacional del Comahue. Resultados: Se describe a Matthiola incana. Se incluyen nombres vulgares y datos sobre distribución, hábitat, ecología y usos. Se presenta una serie de fotografías de campo y bajo microscopio estereoscópico para complementar la descripción. El ensayo de germinación demostró que el 95% de las semillas germina a las 24 hs a 25 °C. Conclusiones: Se reporta por primera vez para Argentina la naturalización de Matthiola incana, única especie del género en el país hasta el momento. Background and aims: The genus Matthiola W. T. Aiton comprises between 50 and 60 species native to the Mediterranean, Central Asia and South Africa. M. incana (L.) W. T. Aiton, indigenous to southern Europe, is naturalized in other regions of the world. In Argentina it is cultivated as ornamental. From floristic observations, a naturalized population was found, which self-perpetuates without human intervention, on the coasts of the Patagonia Argentina. The objective of this work is to report for the first time the naturalization of M. incana in Argentina. M&M: For the identification of the species, the classic methods of taxonomy were followed, specific bibliography was consulted and the collected material was contrasted with digital images of the lectotype. Seeds were collected from the naturalized population and the germinative power was evaluated through a germination test. The collected material that documents the present work was deposited in the ARC herbarium of the Universidad Nacional del Comahue. Results: Matthiola incana is described. The common names and data on distribution, habitat, ecology and uses are included. Photographs are presented to complement the description. The germination test showed that 95% of the seeds germinated after 24 hours at 25°C. Conclusions: The naturalization of Matthiola incana is reported for the first time in Argentina, the only species of the genus in the country until now.

Main conclusion: MiMYB1 and MibHLH2 play key roles in anthocyanin biosynthesis in Matthiola incana flowers. We established a transient expression system using Turnip mosaic virus vector in M. incana. Abstract: Garden stock (Matthiola incana (L.) R. Br.) is a popular flowering plant observed from winter to spring in Japan. Here we observed that anthocyanin accumulation in ‘Vintage Lavender’ increased with flower development, whereas flavonol accumulation remained constant throughout flower development. We obtained five transcription factor genes, MiMYB1, MibHLH1, MibHLH2, MiWDR1, and MiWDR2, from M. incana floral cDNA contigs. Yeast two-hybrid analyses revealed that MiMYB1 interacted with MibHLH1, MibHLH2, and MiWDR1, but MiWDR2 did not interact with any transcription factor. Expression levels of MiMYB1 and MibHLH2 increased in petals during floral bud development. Their expression profiles correlated well with the temporal profiles of MiF3ʹH, MiDFR, MiANS, and Mi3GT transcripts and anthocyanin accumulation profile. On the other hand, MibHLH1 was expressed weakly in all organs of ‘Vintage Lavender’. However, high expression levels of MibHLH1 were detected in petals of other cultivars with higher levels of anthocyanin accumulation than ‘Vintage Lavender’. MiWDR1 and MiWDR2 maintained constant expression levels in petals during flower development and vegetative organs. Transient MiMYB1 expression in 1-month-old M. incana seedlings using a Turnip mosaic virus vector activated transcription of the endogenous anthocyanin biosynthetic genes MiF3ʹH, MiDFR, and MiANS and induced ectopic anthocyanin accumulation in leaves. Therefore, MiMYB1 possibly interacts with MibHLH2 and MiWDR1, and this trimeric protein complex activates the transcription of anthocyanin biosynthetic genes in M. incana flowers. Moreover, MibHLH1 acts as an enhancer of anthocyanin biosynthesis with the MiMYB1–MibHLH2–MiWDR1 complex. This study revealed the molecular mechanism involved in the regulation of anthocyanin accumulation levels in M. incana flowers.

In 2014, glasshouse-grown wasabi (Eutrema japonica) grown in a compost based media displayed symptoms of poor growth and wilting. Visual assessment of the roots showed that 25% of the symptomatic plants sampled had raised black lesions on the roots affecting between 5 and 20% of the total root area. To isolate the causal agent, affected material (approximately 5 mm3) was surface disinfested in sodium hypochlorite (2%) for 30 s, rinsed twice in sterile water and plated on to water agar medium amended with penicillin G (0.2 g/liter) and streptomycin sulfate (0.8 g/liter). Plates were incubated at 20oC until fungal colonies were visible. After three days, colonies of Rhizoctonia solani were identified based on the presence of septate hyphae with right-angle branching, a pure culture was obtained through hyphal tip transfer onto a new plate of PDA. DNA was extracted from a 7-day old plate of the isolate (WAS1) as described previously (Woodhall et al., 2013). The AG of WAS1 was determined as AG2-1 using a subgroup specific real-time PCR assay (Budge et al., 2009b) and confirmed by DNA sequencing as described previously (Lekuona Gomez et al., 2015). The sequence was 100% identical (587/587bp) to a previously identified AG2-1 isolate 1971 (GenBank accession FJ435126) (Budge et al., (2009a). Pathogenicity of the isolate was confirmed by inoculating three healthy one-year-old wasabi plants grown in loam based compost (John Innes No.3) each with four 5 mm fully colonised PDA plugs of isolate WAS1 placed at approx. 40 mm depth in the soil. Four sterile PDA plugs were place in each of three control plants. All six plants were placed in a greenhouse at 21°C, 18h:6h light: dark and watered as required. After 21 days, multiple black root lesions typically 3-5mm in length were observed on the roots of all inoculated plants. No lesions were observed on the control plants. From three lesions per plant, isolations were attempted as described above. Rhizoctonia solani was recovered from all isolations and the resulting cultures all tested positive for AG2-1 using the real-time PCR assay. Isolations were attempted from the roots of healthy control plants but Rhizoctonia was not recovered. Here we demonstrate that R. solani AG2-1 is associated with root necrosis of Eutrema japonica. Rhizoctonia solani AG2-1 has been reported previously in various Brassica crops in the UK (Budge et al., 2009a) and on Matthiola incana (Lekuona Gomez et al., 2015). It has also been reported causing disease in potatoes and as widely present in UK field soils (Woodhall et al., 2013). Although R. solani AG1 and AG4 of R. solani have been reported to infect Eutrema japonica in Japan (Takeuchi et al., 2003; 2008), this is the first finding that identifies AG2-1 as the causal agent. The potential presence of AG2-1 in soil and/or as plant debris should be considered prior to planting susceptible hosts.

Matthiola incana R. Br. (Fam: Brassicaceae) is an ornamental, commonly known as hoary stock has an extremely fragrant flowers, which blooms in dense clusters in a large variety of colors. During a survey of flower nurseries in March 2019 at Indian Institute of Sugarcane Research campus, Lucknow, floral virescence (MiV) symptoms (Fig. 1 A, B) were observed in M. incana pots with an incidence of over 40%. Leaf yellows symptoms were also observed on a weed Acalypha indica (AiLY) in Matthiola nursery (Fig. 1 C). Nested PCR assays were carried out to detect and identify the possible association of phytoplasmas with MiV and AiLY symptoms. Three each of symptomatic MiV and AiLY samples and two non-symptomatic samples were collected and processed for DNA extraction from the leaf midrib by CTAB method. Hishimonus phycitis (HP) (Hemiptera: Cicadellidae) leafhopper feeding on MiV symptomatic plants was also collected and DNA was extracted. The DNA of 8 symptomatic and 4 non-symptomatic plants and from the 10 leafhopper was used as a template for PCR assays. Phytoplasma specific 16Sr RNA gene specific primers (P1/P7 and 3Far/3Rev; Schneider et al. 1995; Manimekalai et al. 2010) and multilocus genes' specific primer pairs for secA (SecAfor1/SecArev3;SecAfo5r/SecARev2; Bekele et al. 2011), secY (SecYF1(VI)/SecYR1(VI);SecYF2(VI)/SecYR1(VI); Lee et al. 2010) and rp genes (rpFIC/rp(I)R1A; rp(VI)F2/ rp(VI)R2; Martini et al. 2007) were employed as previously described. Amplified products of ~1.3kb, ~600bp, ~1.7kb and ~1.0kb of 16S rRNA, secA, secY and rp genes of phytoplasma were consistently amplified in all the MiV and AiLY samples and in the HP leafhopper. No amplifications were achieved in any of the asymptomatic plant samples. Amplified products of all the four genes of MiV, AiLY and HP isolates were purified, sequenced and submitted in GenBank. Sequence comparison and phylogeny analysis of the sequences of the four genes of MiV, AiLY and HP isolates revealed 99% - 100% sequence identity and clustering with clover proliferation phytoplasma related strains (16SrVI group)(Fig.2 A,B,C and D). The virtual RFLP analysis of 17 restriction endonucleases corresponding to the 16S rDNA sequence of MiV, AiLY and HP phytoplasma strains by pDraw program, assigned them into a novel phytoplasma subgroup strain under 16SrVI group, since its HpaII restriction profile was different to earlier classified 16SrVI subgroups but was very close to16SrVI-E subgroup (GenBank acc. no. AY270156) (Fig 3). Earlier, peanut witches' broom (16SrII-A) phytoplasma was identified associated with M. incana from Italy (Davino et al. 2007). However, the association of clover proliferation phytoplasma (16SrVI) related strain associated with virescence symptom of M. incana is the first report in world. The weed (A. indica) and HP leafhopper were also reported as additional hosts of 16SrVI subgroup related new strain in India, which needs further investigation. The report of a new host and new subgroup of clover proliferation phytoplasma related strain in India is having an epidemiological significance and warrants attention.

Clubroot caused by Plasmodiophora brassicae is an increasingly important soilborne disease in China. The host range of P. brassicae was investigated with 30 cruciferous plants, including 16 crop species, 9 ornamentals, and 5 weeds in field and pot-cultured conditions. In the field, 17 species from five genera produced visible galls, and these included radish, Capsella bursa-pastoris, Orychophragmus violaceus, Sinapis alba, and 13 Brassica crops. In pot-cultured conditions, an additional 13 plant species (11 genera) were determined to be hosts of P. brassicae. Five common weeds were found to be hosts of P. brassicae, including C. bursa-pastoris, Lepidium apetalum, Descurainia sophia, S. alba, and Thellungiella salsuginea. The infection of these plants was confirmed via polymerase chain reaction (PCR) with primers specific to P. brassicae. No galls were found on Matthiola incana roots in the field or in pots and no resting spores of P. brassicae were observed in M. incana roots, although P. brassicae was detected in M. incana roots via PCR. Microscopic examination revealed infection only in the root hairs of M. incana roots. These results suggested that M. incana was highly resistant to P. brassicae in China and could be developed as a bait crop. In total, 297 accessions of oilseed rape were tested in the field, and 3 accessions of Brassica napus and 1 accession of B. juncea were found to be highly resistant to clubroot disease. These resistant resources provide options for managing clubroot in P. brassicae-infested fields.

During the spring and fall of 1999, commercial plantings of the cut flower stock (Matthiola incana) in coastal regions of California developed a severe foliar disease. Abaxial leaf surfaces supported the extensive white fungal growth typical of a downy mildew. Adaxial surfaces of newly infected leaves did not show symptoms, but with time, leaves exhibited extensive, diffuse chlorotic areas and slight twisting. Infected plants were not acceptable for the cut flower market and, therefore, were not harvested. Sporangiophores had main trunks with dichotomous branches ending in slender curved tips that were 2.8 μm long. Sporangia were ovoid, measuring 22 to 25 μm long and 19 to 22 μm wide. The fungus was identified as Peronospora parasitica. Pathogenicity was established by gently pressing diseased leaves onto stock plants (cv. Avalanche), incubating plants in a humidity chamber (20 to 22°C, 100% relative humidity) for 48 h, and keeping plants in a greenhouse. Diffuse chlorotic leaf symptoms developed after 10 days, and the same fungus later developed on abaxial surfaces. Uninoculated plants did not develop downy mildew. Inoculations were conducted a second time, and the results were the same. This is the first documentation of P. parasitica on stock in California.

Glucosinolate (GSL) is associated with clubroot disease, which is caused by the obligate biotrophic protist Plasmodiophora brassicae. Due to the complicated composition of GSLs, their exact role in clubroot disease development remains unclear. By investigating clubroot disease resistance in cruciferous plants and characterizing the GSL content in seeds, we can determine if clubroot disease development is related to the components of GSLs. The difference in the infection process between Matthiola incana L. (resistant) and Brassica napus L. (susceptible) was determined. Root hair infection was definitely observed in both resistant and susceptible hosts, but no infection was observed during the cortical infection stage in resistant roots; this finding was verified by molecular detection of P. brassicae via PCR amplification at various times after inoculation. Based on the time course detection of the contents and compositions of GSLs after P. brassicae inoculation, susceptible roots exhibited increased accumulation of aliphatic, indolic, and aromatic GSLs in B. napus, but only aromatic GSLs were significantly increased in M. incana. Gluconapin, which was the main aliphatic GSL in B. napus and present only in B. napus, was significantly increased during the secondary infection stage. Quantification of the internal jasmonic acid (JA) concentration showed that both resistant and susceptible plants exhibited an enhanced level of JA, particularly in susceptible roots. The exogenous JA treatment induced aliphatic GSLs in B. napus and aromatic GSLs in M. incana. JA-induced aromatic GSLs may be involved in the defense against P. brassicae, whereas aliphatic GSLs induced by JA in B. napus likely play a role during the secondary infection stage. Three candidate MYB28 genes regulate the content of aliphatic GSLs identified in B. napus; one such gene was BnMYB28.1, which was significantly increased following both the treatment with exogenous JA and P. brassicae inoculation. In summary, the increased content of JA during the secondary infection stage may induce the expression of BnMYB28.1, which caused the accumulation of aliphatic GSLs in clubroot disease development.

The double flower is one of the most important traits in the floricultural plant Matthiola incana. Although a “doubleness” locus (S/s) was defined by genetic analysis a century ago, the gene responsible for double flowers has not been identified in M. incana. We isolated MiAG from M. incana cultivars, and its sequence and genomic structure were found to be highly similar to the AGAMOUS gene in Arabidopsis. Two independent mutated alleles miag1 and miag2 were identified from the double-flowered individuals of M. incana cultivars. Deletions of 135 bp (from the 2nd exon to the 2nd intron) and 89 bp (from the 7th intron to the 8th exon) were detected in miag1 and miag2, respectively. No transcript was detected in flower buds from miag1 alleles in corresponding cultivars, whereas three mRNA variants with frameshifts were transcribed from the miag2 allele in other cultivars. Thus, two mutated alleles corresponding to the s locus contributed to the ‘eversporting’ type double-flower cultivars in M. incana. Moreover, we also developed co-dominant molecular markers to describe the genotypes of the three alleles of MiAG. Using these DNA markers allows for selection of single- or double-flowered individuals among seedlings that do not display phenotypic differences.

Background For most cruciferous plants, which are known as important crops and a number of weeds, hybrid breeding is hampered by the unavailability of a pollination control system. Male sterility induced by a gametocide can be useful for the utilization of plant heterosis. Results The gametocidal effect of sulfonylurea herbicide tribenuron-methyl was tested across seventeen cruciferous species or subspecies including Brassica juncea, B. carinata, B. oleracea ssp. capitata, B. oleracea ssp. acephala, B. rapa ssp. pekinensis, B. rapa ssp. chinensis, B. rapa ssp. parachinensis, B. nigra, Orychophragmus violaceus, Matthiola incana, Raphanus sativa, Sisymbrium altissimum, Eruca sativa, Sinapis alba, Sinapis arvensis, Capsella bursa-pastoris and Camelina sativa. The plants of 23 cultivars in these species or subspecies were foliar sprayed with 10 ml of 0.2 or 0.4 mg/L of tribenuron-methyl before the vacuolated microspore formed in the largest flower buds; the application was repeated ten to twelve days afterwards. Tribenuron-methyl exposure significantly changed the flowering phenology and reproductive function. The treated plants demonstrated a one to four day delay in flowering time and a shortened duration of flowering, as well as other slight phytotoxic effects including a reduction in plant height and floral organ size. Approximately 80% to 100% male sterility, which was estimated by both pollen staining and selfing seed-set rate, was induced in the plants. As a result, plants were rendered functionally able to out-cross, with an average 87% and 54% manually pollinated seed-set rate compared to the corresponding controls at the 0.2 mg/L and 0.4 mg/L doses, respectively. Conclusions The results suggested that male reproductive function was much more sensitive to tribenuron-methyl exposure than female function. This sulfonylurea herbicide has a promising use as the gametocide for hybrid production in cruciferous plants. Electronic supplementary material The online version of this article (doi:10.1186/s12870-017-1019-1) contains supplementary material, which is available to authorized users.

Typical symptoms of Sclerotinia stem rot were observed in commercial crops of stock in several districts in Korea during the 2012–2013 and 2013–2014 winters. Based on morphological characteristics and phylogenetic analysis of internal transcribed spacer sequences, the pathogen was identified as Sclerotinia sclerotiorum. This is the first report of Sclerotinia sclerotiorum infection of stock in Korea.

The effect of inoculation of two root-colonizing basidiomycete fungi Piriformospora indica and Sebacina vermifera on the host plants from the family Brassicaceae (Brassica oleracea, B. napus, B. nigra, Lepidium sativum, Descurainia sophia and Matthiola incana) was studied in a pot experiment. Both fungi were able to colonize all plant species tested but with different efficiency. Colonized plants reacted with increase of fitness, bigger leaves and more side shoots. Plant heights of five inoculated species were higher than of uninoculated controls. Five species had higher dry weight of shoots and four had higher dry weights of roots. The highest stimulation was stated for B. oleracea and the lowest for L. sativum. It can be connected with the intensity of root colonization, which was greater in B. oleracea. In most cases both fungi influenced growth and dry weight of plants similarly but B. nigra and D. sophia were stimulated more by P. indica and B. napus by S. vermifera.

Anthocyanidin synthase (ANS) is a key enzyme of the proanthocyanidin biosynthetic pathway in Arabidopsis. In this study, we report cloning and characterization of an ANS homolog, BjANS, from Brassica juncea. The BjANS cDNA sequence is 1,377 bp long and has a 1,077-bp open reading frame encoding a deduced polypeptide of 358 amino acids with a predicted molecular weight of 40,862 Da and an estimated isoelectric point of 5.18. The putative BjANS protein shows 91–99% amino acid sequence identity with Arabidopsis thaliana, Matthiola incana, B. rapa and B. oleracea ANS proteins. The expression of BjANS was analyzed in yellow- and black-seeded B. juncea accessions to determine its physiological function. RT–PCR analysis showed that BjANS expressed in the seed coats of the black-seeded lines and in embryos of the all lines, but not in the yellow-seeded seed coats. Proanthocyanidins were also detected by histochemical analysis in the seed coat of black-seeded accessions and in embryos of all the lines, but not the yellow-seeded seed coat. Both transcriptional and histochemical analyses confirmed that BjANS was involved in the biosynthesis of proanthocyanidins and in seed coat color formation of B. juncea. Lack of BjANS expression blocked biosynthesis of proanthocyanidins in the yellow seed coat, and therefore seeds appeared yellow because of transparent testa.

TTG1 (Transparent Testa Glabra 1), a WD-40 repeat protein, is involved in regulation of flavonoid/anthocyanin biosynthesis, seed coat (mucilage) development/pigmentation and trichome formation in leaves. Here, we characterized the TTG1 gene of Matthiola incana wild type (e locus), showing 85.3% similarity to TTG1 of A. thaliana on the nucleotide level and 96.2% on the protein level. A white-flowered and glabrous mutant, line 17, of M. incana exhibits one nucleotide change, leading to an amino acid substitution directly in the WD motif (W158R). Correspondingly, the DFR (dihydroflavonol 4-reductase) gene, in which the expression is known to be dependent on TTG1, is not expressed in Matthiola mutant lines 17 (and 19). Comparison of the GC content of the Matthiola TTG1 (54.1%) and Arabidopsis TTG1 (46.1%) genes revealed a strong difference, mostly obtained by neutral substitutions (C to T transitions). To examine whether this is an ecologically influenced trend, a fragment of TTG1 was characterized from another Matthiola species (M. tricuspidata) and from Malcolmia flexuosa subsp. naxensis from the eastern Mediterranean, near a beach with sandy and salty soils. Both Matthiola species have a higher GC content in the TTG1 gene than Arabidopsis and the closer-related Malcolmia, indicating that the GC content is rather an evolutionary than an ecological signal. A similar WD-40 repeat protein gene (containing no intron in the 3' untranslated region) with high similarity to the Arabidopsis TTG1-like (AtAN11) gene was found in Matthiola.

Flavanone 3-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDHA probe from Dianthus cavophyllus was used to isolate FHT-encoding cDHA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5` leader region and 288 bp 3` untranslated region. Comparison of the coding region of this FHT cDHA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRHA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

Biosynthesis is well elucidated for 5-hydroxyflavonoids (phloroglucinol type), but for 5-deoxyflavonoids (resorcinol type) the knowledge is still limited. We provide detailed and optimized protocols for the synthesis of (14C)-labeled 6′-deoxychalcones, 5-deoxyflavanones, 5-deoxydihydroflavonols and 5-deoxyleucoanthocyanidins. With the exception of the formation of 6′-deoxychalcones, all steps were performed enzymatically using enzymes normally involved in the formation of 5-hydroxyflavonoids. The availability of (14C)-labeled substrates will facilitate future work on the hitherto largely unknown biosynthesis of 5-deoxyflavonoids. In particular, the 5-deoxyleucoanthocyanidins, which are more stable than the corresponding 5-hydroxy compounds, may provide excellent tools for investigating enzymes, which use the unstable 5-hydroxyleucoanthocyanidins as natural substrates. As a first example, the conversion of (14C)-labeled 5-deoxyleucoanthocyanidins to dihydroflavonols in the presence of NADP+ was shown. Studies with defined genotypes of Matthiola incana possessing or lacking dihydroflavonol 4-reductase activity and genetically modified yeast expressing the Matthiola enzyme confirmed that the reaction is catalyzed by the well-known dihydroflavonol 4-reductase, which catalyzes the conversion of dihydroflavonols to leucoanthocyanidins (forward reaction). Thus, the reverse reaction of dihydroflavonol 4-reductase could be demonstrated for the first time. The forward reaction shows an optimum at pH 6.25, the reverse reaction at pH 7.75. The impact of the results on the regulation of flavonoid accumulation is discussed.

Basic helix-loop-helix (bHLH) proteins, similar to mammalian Myc transcription factors, regulate the anthocyanin biosynthetic pathway in both monocots and dicots. Two Arabidopsis bHLH genes, GLABRA3 (GL3) and MYC-146, encode proteins that are similar throughout the predicted amino acid sequence to R and DELILA, which regulate anthocyanin production in maize and snapdragon, respectively. Northern blot analysis indicates that MYC-146 is most highly expressed in flower buds and flowers. Expression of a MYC-146 cDNA from the CaMV 35S promoter was unable to complement the anthocyanin deficiency in a ttg1 mutant of Arabidopsis and resulted in no obvious phenotypic change in Columbia plants. However, transient expression of GL3 and MYC-146 upon microprojectile bombardment of petals of a white-flowered mutant of Matthiola incana was able to complement anthocyanin deficiency. The lack of anthocyanin-deficient Arabidopsis mutants mapping to the locations of GL3 and MYC-146 suggests that the two bHLH proteins may be partially redundant and overlap in function.

Recently, it was found that stem elongation and flowering of stock Matthiola incana (L.) R. Br. are promoted by exogenous gibberellins (GAs), including GA4, and also by acylcyclohexanedione inhibitors of GA biosynthesis, such as prohexadione-calcium (PCa) and trinexapac-ethyl (TNE). Here, because it was unclear how GA biosynthetic inhibitors could promote stem elongation and flowering, their effect on GA biosynthesis has been examined by quantifying endogenous GA levels; also, the sensitivity of stem elongation and flowering to various GAs in combination with the inhibitors was examined. Stem elongation and flowering were most effectively promoted by GA4 when combined with PCa and, next in order, by 2,2-dimethyl-GA4, PCa, GA4+TNE, TNE, GA9+PCa and by GA4. There was little or no promotion by GA1, GA3, GA9, GA13, GA20 and 3-epi-2,2-dimethyl-GA4. Both the promotive effects of the acylcyclohexanediones on stem elongation and flowering, particularly when applied with GA4, and the fact that TNE caused a build-up of endogenous GA4 imply that one effect of TNE at the lower dose involved an inhibition of 2β-hydroxylation of GA4 rather than an inhibition of 20-oxidation and 3β-hydroxylation of GAs which were precursors of GA4. Overall, these results indicate that: (1) GAs with 3β-OH and without 13-OH groups (e.g. GA4) are the most important for stem elongation and flowering in M. incana; (2) growth promotion rather than inhibition can result if an acylcyclohexanedione acts predominantly to slow 2β-hydroxylation and so slows inactivation of active gibbberellins, including GA4. It follows that a low dose of an acylcyclohexanedione can be a ‘growth enhancer’ for any applied GA that is liable to inactivation by 2β-hydroxylation.

Shoots and flower buds of stock (Matthiola incana (L.) R. Br. cv. Sourei) were analysed for gibberellin (GA). After purifying the acidic ethylacetate-soluble fraction by several chromatographic procedures, GA fractions were surveyed by GC-MS and Kovats retention indices (Rl). Consequently, GA1, GA4, 3-epi-GA4, GA8, GA19, GA20, GA24, GA34, GA37 and GA53 were identified. A further GA-like substance was detected and shown to be 12β-OH-GA12 (GA112) by comparison with an authentic compound prepared by synthesis.

To determine the developmental patterns of arbuscular mycorrhizae (AM) in three crucifers (Brassicaceae) of differing life histories, we inoculated seedlings of the annual Capsella bursa-pastoris, biennial Hesperis matronalis, and the perennial Matthiola incana with Glomus intraradices. The plants were grown either alone or in a matrix of living roots of the mycotrophic grass Sorghum sudanense. The percent root length colonized was greatest in C. bursa-pastoris and least in H. matronalis. Colonization was greater in plants grown in the grass matrix than in plants grown alone, and colonization in grass matrix-grown plants continued to increase over the 90-day growth period, whereas colonization leveled off or decreased near the end of the growth period in crucifers grown alone. No arbuscules were observed in crucifer roots at any time, which suggests that AM in these crucifers is nonfunctional. Furthermore, the increase in colonization only in pots with both crucifers and active mycotrophic roots suggests that AM development in crucifer roots is primarily the consequence of progressive root senescence in the crucifer and continued inoculum spread from the mycotrophic plant.

Ten acylated pelargonidin 3-sambubioside-5-glucosides were isolated from the red-purple flowers of Matthiola incana, and also pelargonidin 3-glucoside was isolated from the brownish-red flowers of this plant. FAB mass measurements of 10 acylated anthocyanins gave their molecular ions [M]+ at 903-1195 m/z, which were based on acylated pelargonidin 3-sambubioside-5-glucosides with malonic acid, sinapic acid, p-coumaric acid, caffeic acid and/or ferulic acid. This was confirmed by the analysis of NMR spectra and the experiments of acid and alkaline hydrolysis. By spectral and chemical methods, seven of the 10 pigments were determined to be pelargonidin 3-O-[2-O-(2-O- (acyl-I)-β- d -xylopyranosyl )-6-O- (acyl-II)-β- d -glucopyranoside ]-5-O-[6-O- (malonyl)-β- d - glucopyranoside ] , in which acyl moieties varied between sinapic, ferulic, caffeic and p-coumaric acids. The occurrence of these pigments was examined in 10 red-purple, 10 salmon-pink, three apricot and three copper colour cultivars of M. incana by HPLC. The acylated pelargonidin 3-sambubioside-5-glucosides were present as the dominant pigments in the red-purple, salmon-pink and apricot colour cultivars. On the other hand, pelargonidin 3-glucoside was present as a dominant anthocyanin in the copper colour cultivars and also pelargonidin 3-sambubioside-5-glucoside was confirmed by HPLC as a minor pigment in the copper colour flowers.

The inheritance of flowering time (FT) in a cross between early-flowering (P1) and late-flowering (P2) Matthiola incana genotypes, was investigated. The distribution of FT in F1, F2 and BC1 generations indicated an additive genetic control with partial dominance of earlines alleles, particularly with regard to double-flowered plants. Single-flowered plants flowered earlier than double-flowered plants, averaged within both parental lines; however, the mean difference between singles and doubles was 16 days for P1 as compared with 6 days for P2. This flower-doubleness related delay in FT was found to be heritable, by analysis of F3 family means. The implications of these results on the breeding of stock cultivars is discussed.

The inheritance and linkage relationships of a leaf morphology gene of Matthiola incana were investigated. The allele for sinuate leaf shape, c, was found to be recessive to the allele for normal entire leaf, C. The c allele was tightly linked to the recessive allele for double flowering, s. The recombination frequency between the two loci was close to zero. The mode of inheritance of the C gene was in accordance with the hypothesis that a pollen lethal gene is responsible for the constant 1:1 segregation ratio of double-flowered (= male sterile) to single-flowered (= fertile) plants in most M. incana breeding lines. The sinuate leaf allele seemed to reduce the frequency and delay the flowering of double-flowered plants. The importance of the C gene as a double flowering marker in the cultivation and the breeding of M. incana is discussed.

The fatty acid composition of seed-oil of breeding lines and F1 hybrids of Matthiola incana was analyzed, using direct esterification and gas chromatography. The breeding lines tested differed significantly with respect to the levels of palmitic, oleic, linoleic and linolenic acids. Embryonic-stage heterosis in linolenic acid concentration was demonstrated by F1 hybrid seeds, derived from mating horticulturally different lines of M. incana. Linolenic acid content was negatively correlated with both oleic acid content (r=−0.85) and linoleic acid content (r=−0.66). None of the breeding lines or the F1 hybrids significantly passed the limit of 67% linolenic acid. Possible genetic and biochemical explanations for the above phenotypic data are discussed.

Rhizoctonia solani is a species complex consisting of 13 different anastomosis groups (AGs) and numerous subgroups. Each AG or subgroup is usually associated with a particular host. In 2014, stock plants ( Matthiola incana , Brassicaceae)…

Cotton fibres are single, highly elongated cells derived from the outer epidermis of ovules, and are developmentally similar to the trichomes of Arabidopsis thaliana. To identify genes involved in the molecular control of cotton fibre initiation, we isolated four putative homologues of the Arabidopsis trichome-associated gene TRANSPARENT TESTA GLABRA1 (TTG1). All four WD-repeat genes are derived from the ancestral D diploid genome of tetraploid cotton and are expressed in many tissues throughout the plant, including ovules and growing fibres. Two of the cotton genes were able to restore trichome formation in ttg1 mutant Arabidopsis plants. Both these genes also complemented the anthocyanin defect in a white-flowered Matthiola incana ttg1 mutant. These results demonstrate parallels in differentiation between trichomes in cotton and Arabidopsis, and indicate that these cotton genes may be functional homologues of AtTTG1.

Leaf pigments, such as chlorophyll and carotenoids, are essential plant molecules. They provide carbohydrates and energy during all plant life. Leaf pigments are also important parameters of decorative plants, such as floriculture items, cut foliage and flowers. Leaf yellowing is a form of senescence caused by an internal hormone imbalance, such as a lack of cytokinins. The aim of this study was to investigate the changes in total carotenoids and endogenous ABA in cut flower stock leaves during post-harvest life. The effect of pulse treatment with 5 or 10 μM thidiazuron (TDZ), 150 mg l−1 8-hydroxyquinoline sulphate (8-HQS) and combinations of TDZ with 8-HQS on total carotenoids and ABA concentration was also investigated. Results showed that total carotenoids drastically decreased from 1548 μg cm−2, until reaching 565 μg cm−2 at the end of vase life. Endogenous ABA strongly increased at the same time, going from 167 ng g−1 DW at the beginning of the experiment to 1322 ng g−1 DW at the end of vase life. The TDZ inhibited carotenoid degradation, but did not affect the ABA concentration, while the 8-HQS did not prevent carotenoid degradation and the ABA concentration was only slightly affected. ABA seems to be a secondary senescence bio-product that may have a synergic effect with other senescence inducers dramatically accelerating leaf senescence.

The major anthocyanin in the leaves and stems of Arabidopsis thaliana has been isolated and shown to be cyanidin 3-O-[2-O(2-O-(sinapoyl)-beta-D-xylopyranosyl)-6-O-(4-O-(beta-D-glucopyranosyl)-p-coumaroyl-beta-D-glucopyranoside] 5-O-[6-O-(malonyl) beta-D-glucopyranoside]. This anthocyanin is a glucosylated version of one of the anthocyanins found in the flowers of the closely related Matthiola incana.

Seeds from 15 Cruciferae species, native to Israel, were collected and examined for fatty acids in order to identify species rich in fatty acids with potential for industrial use from the wild gene collection. In species rich in erucic acid (22:1), such as Crambe and Sinapis a low content of polyunsaturated linolenic acid (18:3) was found; the same negative correlation was observed in species low in erucic acid ( Matthiola , Lepidium ), in which 64 and 42% linolenic acid, respectively was found. No correlation was detected between the content of erucic and oleic acids, as opposed to the situation in cultivated rape seeds. Plants with different oil compositions were found in the same natural habitat. Fatty acid profiles could not be related to ecological conditions.

Flower color was modified in forsythia (Forsythia x intermedia cv ‘Spring Glory’) by inducing anthocyanin synthesis in petals through sequential Agrobacterium-mediated transformation with dihydroflavonol 4-reductase from Antirrhinum majus (AmDFR) and anthocyanidin synthase from Matthiola incana (MiANS) genes. This is the second report of flower color modification of an ornamental shrub after rose, and the first time an ANS gene is used for this purpose. Double transformants (AmDFR+MiANS) displayed a novel bronze-orange petal color, caused by the de novo accumulation of cyanidin-derived anthocyanins over the carotenoid yellow background of wild type (wt), and intense pigmentation of vegetative organs. Transformation with single genes (either AmDFR or MiANS) produced no change in flower color, showing a multistep control of late anthocyanin pathway in petals of forsythia. Analysis of relevant late flavonoid pathway genes – an endogenous flavonoid glycosyltransferase (FiFGT) and transformed DFR and ANS genes – showed appropriate expression in flower organs. Functional characterization of FiFGT expressed in E. coli revealed its ability to metabolize both flavonols and anthocyanidin substrates, a prerequisite for effective anthocyanin accumulation in petals of plants transformed with constructs leading to anthocyanidin synthesis. Biochemical analyses of flavonoid compounds in petals and leaves showed that, besides anthocyanin induction in petals of double transformants, the accumulation pattern of flavan-3-ols was quantitatively and qualitatively modified in petals and leaves of transformants, in agreement with the most recent model proposed for flavan-3-ol synthesis. On the other hand, phenylpropanoid, flavone and flavonol pools were not quantitatively affected, indicating a tight regulation of early flavonoid pathway.

A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of three lines of Matthiola incana is described. Protoplasts were isolated from leaves of 21-28 days old Matthiola plants grown in controlled environment. Sustained divisions were achieved when protoplasts were embedded in sodium alginate. Up to 2.0 % of the protoplasts developed into colonies which could be transferred to shoot regeneration media. More than 25 % of the obtained calluses regenerated shoots. About 4 % of these shoots could be rooted and after transfer to soil phenotypically normal plants have been obtained.

A disease of stock (Matthiola incana) in Australia, caused by Pseudomonas syringae pv. maculicola, is described for the first time. The pathogen is systemic, causing vascular necrosis and progresses basipetally from infected flowers. Bacteria were isolated from seed produced from inoculated blossoms; however, transmission to seedlings grown from infected seed was not proven. The symptoms differ from those of a disease reported to be caused by P. syringae pv. syringae.

A cDNA clone (pcM12) of the chalcone synthase (CHS) of Matthiola incana R. Br. (Brassicaceae) was isolated from a cDNA library, sequenced and analysed. It comprises the complete coding sequence for the CHS and 5' and 3' untranslated regions. The deduced amino acid sequence shows that the Matthiola incana CHS consists of 394 amino acid residues. Comparison with CHS amino acid sequences of other plants indicates more than 82% homology.

Accessions ofM. tricuspidata (L.) R. Br.,M. aspera Boiss. andMatthiola longipetala (Vent.) DC. were tested for their reaction to bacterial blight of stock (M. incana R. Br.), caused byXanthomonas campestris pv.incanae. Potted plants were sprayed with bacterial cell suspension, adjusted to 108 cells/ml. The tested accessions of the three species were found highly resistant, while control plants ofM. incana were severely infected. InterspecificF 1 hybrid plants betweenM. incana andM. tricuspidata also exhibited high resistance to the disease. The implications and the expected difficulties in breeding stock for bacterial blight resistance are discussed.

After application of dihydroflavonols, naringenin, or suitable substituted chalcones, anthocyanins were synthesized in three genetically defined acyanic lines of Matthiola incana, indicating that the corresponding genetic block concerns the synthesis of the chalcone-flavanone intermediate. Independent of the precursors used, only cyanidin derivatives were produced. This supports the hypothesis that the oxygenation pattern of the B ring in anthocyanin formation is determined at a stage of a C15 intermediate. In addition to the gene responsible for the oxygenation of the 3' position, the genes responsible for the glycosylation in the 3 and 5 positions of the anthocyanin molecule, and those responsible for the acylation with various hydroxycinnamic acids can still exert their influence. Two further genetically defined lines containing flavonol glycosides were not able to synthesize anthocyanins with any of the precursors tested. Their genetic blocks are assumed to be localized after dihydroflavonol synthesis but before anthocyanin formation.

Petals of genetically defined lines of Matthiola incana R.Br. contain a glycosyltransferase which catalyzes the transfer of the xylosyl moiety of uridine 5'-diphosphate-xylose to the glucose of cyanidin 3-glucoside. The enzyme also uses 3-glucosides of pelargonidin and delphinidin, cyanidin 3-(p-coumaroyl)-glucoside and 3-(caffeoyl)-glucoside as substrates. The xylosyltransferase exhibits a pH optimum of 6.5. The enzyme activity depends on the stage of bud and flower development. Accumulation of cyanidin 3-glucoside during flower development is correlated with xylosyltransferase activity.

Summary Some specific and subspecific new entities are described in this note: Petrorhagia rupestris Brullo et Furnari, sp. nov., Sedum cyrenaicum Brullo et Furnari, sp. nov., Antirrhinum gebelicum Brullo et Furnari, sp. nov., Matthiola incana (L.) R. Br. ssp. cyrenaica Brullo et Furnari, subsp. nov., Sedum laconicum Boiss. et Heldr. ssp. pentapolitanum Brullo et Furnari, subsp. nov., Erica sicula Guss. ssp. cyrenaica Brullo et Furnari, subsp. nov., Plantago lagopus L. ssp. ptolemaidis Brullo et Furnari, subsp. nov., Tolpis virgata (Desf.) Bert. ssp. apolloniae Brullo et Furnari, subsp. nov. Beside the nomenclature and the taxonomie rank of other entities is changed: Salsola sieberi Presl ssp. cyrenaica (Maire et Weiller) Brullo et Furnari, stat. nov., Herniaria glabra L. ssp. cyrenaica (Pamp.) Brullo et Furnari, stat. nov., Frankenia syrtica (Maire et Weiller) Brullo et Furnari, comb, et stat. nov., Matthiola incana L. R. Br. ssp. pulchella (Tin. ex Guss.) Brullo et Furnari, comb, et stat. nov., Rhamnus ...

A global collection of 123 putative isolates of Fusarium oxysporum from crucifers was examined for pathogenicity, isozyme polymorphism, and vegetative compatibility. Of these isolates, 103 were found to be pathogenic on one or more of six differential crucifer cultivars. Three patterns of isozyme polymorphism (electrophoretic types) were found and by means of a nitrate reductase complementation test, three major vegetative compatibility groups were identified that could differentiate among the F. oxysporum pathotypes. Complete correspondence was found among pathotype, electrophoretic type, and vegetative compatibility. It seems appropriate to classify isolates from the Cruciferae into the subspecific taxa, F. oxysporum f.sp. conglutinans, F. oxysporum f.sp. raphani, and F. oxysporum f.sp. matthioli, based on their naturally infected host species, Brassica oleracea, Raphanus sativus, and Matthiola incana, and on estimates of genetic identity. Within formae speciales, races can be identified based on intraspecific host specialization. Geographic origin was not found to be associated with the vegetative compatibility, isozyme phenotype, or pathotype. Isozyme polymorphisms also differentiated among four F. oxysporum formae speciales from other host families and among various Fusarium species.

(+)Leucopelargonidin [(2R,3S,4R)-3,4,5,7,4'-pentahydroxyflavan] and (+)leucocyanidin [(2R,3S,4R)-3,4,5,7,3',4'-hexahydroxyflavan] were synthesized from (+)dihydrokaempferol and (+)dihydroquercetin, respectively, by sodium-borohydride reduction. The chemical and optical purity of these compounds was established by ultraviolet, proton-nuclear-magnetic-resonance, and optical-rotatory-dispersion spectroscopy. Supplementation experiments with these leucoanthocyanidins were carried out with genetically defined acyanic flowers of Matthiola incana. Feeding of leucopelargonidin and leucocyanidin to line 17 (blocked between dihydroflavonols and anthocyanins) and line 18 (blocked in the chalcone-synthase gene) led to formation of the corresponding anthocyanidin 3-O-glucosides, whereas supplementation of line 19 (blocked in a locus other than line 17 between dihydroflavonols and anthocyanins) did not result in anthocyanin synthesis. The conversion of leucopelargonidin into pelargonidin 3-O-glucoside was further confirmed by incorporation of [4-(3)H]leucopelargonidin into pelargonidin derivatives. The results are strong indications for the role of leucoanthocyanidins (flavan-3,4-diols) as intermediates in anthocyanin biosynthesis.

A cell-free extract from flowers of Matthiola incana catalyzes a NADPH-dependent stereospecific reduction of (+)-dihydrokaempferol to 3,4-cis-leucopelargonidin (5,7,4'-trihydroxyflavan-3,4-cis-diol). The pH-optimum of this reaction is around 6. The rate of reaction with NADH was about 50% of that found with NADPH. (+)-Dihydroquercetin and (+)-dihydromyricetin were also reduced by the enzyme preparation to the corresponding flavan-3,4-cis-diols. Correlation between the genotype of M. incana and the presence of dihydroflavonol 4-reductase is strong evidence that this enzyme is involved in anthocyanin biosynthesis.

The expression of the key enzyme of flavonoid biosynthesis, chalcone synthase (CHS), has been followed in different genotypes of Matthiola incana R.Br. (Brassicaceae) which are genetically defined with respect to anthocyanin production. Enzyme activity was determined by a radioactive assay in crude flower extracts. The amount of enzyme protein in the developing flower was determined by use of SDS-PAGE, protein blotting, reaction with an antiserum against CHS of parsley (Petroselinum hortense), and PAP staining. The molecular weight of about 41 500 of the CHS subunits corresponds with that obtained from other higher plants. Steps of flower development were subdivided into stages-1,0, I-IV. During flower development of a Matthiola line with coloured petals (line 07) a defined pattern of CHS enzyme production can be observed: At the stage of bud opening (stage 0-I) a dramatic increase of the amount of CHS enzyme prodein in the petals occurs. This is quite different from results obtained with petals of the white flowering mutant line 18 bearing a genetic defect in the gene f coding for CHS. Here a reduced and nearly constant level of CHS enzyme protein can be observed during flower development. This line is most attractive for our studies of the regulation of enzyme synthesis because under stress conditions a slight colouring of the flower petals occurs, which is uniformly distributed and line-specific. This suggests that we are dealing with a CHS mutant producing a rather inactive enzyme protein at a low level. This protein may regain enzyme activity under certain environmental conditions. Preliminary investigations suggest a rather high level of CHS mRNA transcription at the bud opening stage of the flowers. Other white flowering mutant lines, line 17 (genotype ee) and line 19 (gg) with a late block in the flavonoid biosynthesis pathway, exhibit a concomitant reduction of CHS enzyme activity and protein content in comparison to anthocyanin-producing lines with the f(+)f(+)e(+)e(+)g(+)g(+)-genotype.

Anthocyanidin 3-glucosides and 3-sambubiosides acylated with 4-coumarate or caffeate were identified in flower extracts of lines of Matthiola incana with wild-type alleles of the gene u. An enzyme activity was demonstrated catalysing the acylation of 3-glucosides and 3-sambubiosides with 4-coumarate or caffeate using the respective CoA esters as acyl donors. Anthocyanins glycosylated in both the 3- and 5-positions were not acylated. The enzyme exhibited an pH optimum at 6.5 and was inhibited by divalent ions, EDTA, diethylpyrocarbonate and p -chloromercuribenzoate. Accumulation of acylated 3-glycosides during bud development is correlated with acyltransferase activity. In confirmation of the chemogenetic work, acyltransferase activity was found only in enzyme extracts from flowers of lines with the wild-type allele u + .

Downy mildew (Peronospora matthiolae (Roumeguerre) Gaumann) of stock, which was first found in New Zealand in 1958, became a serious and widespread disease within a year. Life history studies of the fungus indicated that it produced only asexual conidia, and that it was not systemic in the plant. Amongst the Cruciferae the fungus infected only species of M. atthiola, both M. incana and M. bicornis being susceptible. All varieties of stock tested were susceptible, but there was evidence that intensity of sporulation varied between varieties. Under optimum conditions the incubation period of the fungus was 5–6 days. It required high humidities for sporulation, and the optimum temperature for infection and sporulation was 15.5°–21°c. Streptospray (500 p.p.m. + glycerol, 1 per cent) protected plants from infection, and also acted as an eradicant. Sprays containing ferbam, cuprous oxide emulsion, Phaltan, dichlone, U.C.P. 21, Trioneb, and zineb gave protection but were not eradicant. Best control of n...

The anthocyanins present in colour varieties of Lathyrus odoratus and Streptocarpus have been identified; the sugars of some forty other anthocyanins have also been examined. In all, twenty-two new anthocyanins have been found; they include the first representatives of the following glycosidic classes: 3-rhamnosides, 3-(xylosylgalactosides), 5-glucoside-3-rhamnosides, 5-glucosides-3-sophoroside, 3-sophoroside-7-glucosides and 5-glucoside-3-sambubiosides. Each of the two unusual disaccharides sophorose and sambubiose occurs combined in the anthocyanins of several plant genera. Structures previously proposed for pigments of Matthiola incana flowers and of red cabbage have been revised. This work brings the number of known classes of anthocyanins to seventeen.