12-h Photoperiod Research Articles

LED lighting affected the growth and metabolism of eggplant and tomato transplants in a greenhouse

Light-emitting diodes (LEDs) were used for the spring greenhouse cultivation of eggplant (Solanum melongena L.) ‘Milar F1’ and tomato (S. lycopersicon L.) ‘Benito F1’ transplants. Seedlings were grown under natural light conditions with the supplemental LED light. A 16-h photoperiod provided plants with a DLI of 12.6 (eggplant) and 9.6 (tomato) mol m2/day. Four supplemental light spectra were tested: L1 (90% red + 10% blue); L2 (80% red + 20% blue); L3 (43% red + 42% blue+15% green) and L4 (56% red + 26% blue + 15% green + 3% UV-A). The PPFD in each LED light treatment was 150 ± 20 µmol/m2·s. Compared to the control plants (without LED lighting), the eggplant transplants had about a 25% larger leaf area and a higher level of total phenol content as well as a higher antiradical scavenging activity under the L1 spectrum. The favourable spectrum for the tomato transplants consisted of red to blue in a ratio of 1 : 1 mixed with a green light (L3) – the leaves were characterised by a higher content of dry matter, soluble sugars, photosynthetic pigments and total phenols; also the radical scavenging activity increased in comparison to the control group. It was shown that the supplemental irradiation of transplants was economically acceptable.

Irradiance-driven 20-hydroxyecdysone production and morphophysiological changes in Pfaffia glomerata plants grown in vitro.

Pfaffia glomerata possesses potential pharmacological and medicinal properties, mainly owing to the secondary metabolite 20-hydroxyecdysone (20E). Increasing production of biomass and 20E is important for industrial purposes. This study aimed to evaluate the influence of irradiance on plant morphology and production of 20E in P. glomerata grown in vitro. Nodal segments of accessions 22 and 43 (Ac22 and Ac43) were inoculated in culture medium containing MS salts and vitamins. Cultures were maintained at 25 ± 2°C under a 16-h photoperiod and subjected to irradiance treatments of 65, 130, and 200μmolm-2s-1 by fluorescent lamps. After 30days, growth parameters, pigment content, stomatal density, in vitro photosynthesis, metabolites content, and morphoanatomy were assessed. Notably, Ac22 plants exhibited 10-fold higher 20E production when cultivated at 200μmolm-2s-1 than at 65μmolm-2s-1, evidencing the importance of light quantity for the accumulation of this metabolite. 20E production was twice as high in Ac22 as in Ac43 plants although both accessions responded positively to higher irradiance. Growth under 200μmolm-2s-1 stimulated photosynthesis and consequent biomass accumulation, but lowered carotenoids and anthocyanins. Furthermore, increasing irradiance enhanced the number of palisade and spongy parenchyma cells, enhancing the overall growth of P. glomerata. Graphical abstract.

First Report of Powdery Mildew Caused by Podosphaera xanthii on Ixeris denticulata in China.

Ixeris denticulata (Houtt.) Stebb is an annual herbaceous plant in the family of Asteraceae, which is native to Europe or central Asia. This plant is widely distributed in China and is commonly used for edible and medicinal purposes. In February 2019, typical symptoms of powdery mildew were observed on 70% of I. denticulata plants on the campus of Hainan University (20° 3' 25″ N; 110° 19' 4″ E) in Haikou, Hainan Province, China. White, superficial mycelia and conidia covered the leaf surfaces of affected plants, resulting in leaf curling, discoloration and defoliation. Hyphal appressoria were nipple-shaped, and solitary. Conidiophores were straight, cylindrical, 109 to 259 × 9 to 16 µm (n = 50), and produced 3 to 5 immature conidia in chains with a crenate outline. Foot cells were cylindrical, straight or sometimes constricted at the basal septum, 30 to 62 µm long (n = 100). Conidia were ellipsoid-ovoid to doliiform, 23 to 33 × 15 to 23 µm (n = 100) with a length/width ratio of 1.1 to 1.9, with well developed fibrosin bodies, and produced germ tubes from the lateral position. Based on these morphological characteristics, this pathogen was provisionally identified as Podosphaera xanthii (Braun and Cook 2012). The teleomorph was not detected. A specimen was deposited in the Hainan University Plant Pathology Herbarium as HNID-18. In order to confirm the identification, genomic DNA was extracted from mycelium and conidia collected from a single leaf using a fungal DNA kit (Omega Bio-Tek, USA). The rDNA internal transcribed spacer (ITS) region and 28S rDNA were amplified with the primer pairs ITS1 and ITS4 (White et al. 1990) and sequenced directly. The resulting 577-bp sequence was deposited in GenBank (Accession No. MT739423). The GenBank BLAST analysis of the ITS sequence showed 100% similarity with P. xanthii on Bidens sp. from Thailand (LC270780), as well as with P. xanthii from Eclipta prostrata (MT260063) and Cyanthillium cinereum (MN203658) from China. Additionally, the 28S rDNA region was amplified using the primer pairs NL1 and NL4 (O'Donnell 1993, Accession No. MT739424). The amplicon was sequenced in both directions and shared 100% similarity with P. xanthii (MK357436, LC371333 and MH137264). To fulfill Koch's postulates, five healthy potted plants of I. denticulata were inoculated by gently pressing a powdery mildew-infected leaf onto 15 young leaves. Five non-inoculated plants served as controls. All plants were maintained in a greenhouse at 24 to 30°C, 70% relative humidity and a 16-h photoperiod. After 7 days, inoculated leaves showed signs and symptoms of powdery mildew whereas no signs or symptoms were observed on the control plants. The fungus observed on the inoculated plants was identical morphologically to that on the originally infected leaves. Powdery mildew of I. chinensis caused by Golovinomyces sonchicola has been reported previously from Korea (Choi et al. 2014). Recently, P. xanthii was also shown to infect Ixeridium dentatum in Korea (Lee and Nguyen 2018). To our knowledge, this is the first record of P. xanthii infecting I. denticulata in China. We are concerned that the pathogen will cause severe damage and affect the yield and quality of the host, and even pose a threat to I. denticulata in the future.

First report of Sclerotinia blight caused by Sclerotinia sclerotiorum on Pinellia ternata in China.

Pinellia ternata is a perennial herb that belongs to the Araceae Family. The tuber of the P. ternata plant contains active ingredients such as alkaloids and pinellia starch, and can be used as a Chinese medicine for antineoplastic and anti-inflammatory disorders in humans (Marki et al. 1987). In April 2019, lesions with rotting were observed during the flowering phase on spathes of P. ternata in a field in Qianjiang City (30°50'N, 112°92'E), Hubei Province, which is one of the main production areas of P. ternata in China. Approximately 10% of P. ternata plants were symptomatic. The initial symptoms of infection were reddish-brown lesions, followed by the appearance of white cottony mycelia. Subsequently, lesions became gray-brown, and rotted with white mycelium that eventually formed on the lesions after 2 to 3 days. In the later phases, spathes were completely rotted and mycelia began to spread to the stems, until the plant wilted and died. Ten spathes at the initial stage of infection were collected in Zhugentan Town, Qianjiang City disinfested with 0.5% sodium hypochlorite for 1 min and 75% alcohol for 20 sec, then washed with sterile distilled water three times, dried, and placed on Petri plates with potato dextrose agar (PDA) and incubated at 22℃ for two days. Six fungal isolates were obtained and purified by hyphal tip isolation in fresh culture, respectively. Culture media was covered with white hyphae after 3 to 4 days of incubation, and dark-gray, rough, irregular sclerotia (1.5 to 5.5 mm in length × 1.0 to 3.5 mm in width) formed on the margins of the media, followed by the melanization as the culture aged. Eventually, black sclerotia were formed and wrapped by white hyphae. All isolates were preliminarily identified as Sclerotinia sclerotiorum (Lazarovits et al. 2000). To further identify the pathogens, molecular identification was performed with one of the six isolates (BXH1). Polymerase chain reaction was performed with primers ITS1/ITS4 for the internal transcribed spacer (ITS) region (White et al. 1990) and primers SSasprF/SSasprR for the aspartyl protease gene (Abd-Elmagid et al. 2013). BLAST search analysis revealed that the 456-bp ITS sequence (GenBank MT436756.1) was ≥99% similar to S. sclerotiorum (MT177216.1, MN105884.1, MG931017.1, etc.), and the 173-bp aspartyl protease gene sequence (GenBank MT584031.1) was too (MK028159.1, MK028161.1, AF271387.1, etc.). Pathogenicity tests were carried out by inoculating disease-free, surface-disinfested spathes of thirty 30-day-old P. ternata plants in plastic pots with a sterilized mixture of peat moss and vermiculite (3:1). Five mycelial plugs (6 mm) were excised from the margin of a colony cultured for 5 days. The plugs were placed on five spathes covered with wet sterile cotton at 22±1℃, and 80% relative humidity, with a 12-h photoperiod. Five control plants were inoculated with noncolonized PDA plugs. Lesions formed on the second day, then rot and white hyphae began to appear on the third day, while the controls had no symptoms. Similar results were obtained in three repeated experiments with S. sclerotiorum being re-isolated from all diseased plants, in accordance with Koch's postulates. This disease is an emerging problem in P. ternata fields in Qianjiang, leading to extensive yield reduction and significant economic losses. To our knowledge, this is the first report of Sclerotinia blight on P. ternata in China.

First Report of Fusarium poae Causing Fusarium Head Blight of Wheat in Georgia, USA.

Fusarium head blight (FHB) is one of the most troublesome fungal diseases challenging US wheat (Triticum aestivum L.) production (Savary et al. 2019). Harmful mycotoxin contamination, primarily due to deoxynivalenol (DON) in the Fusarium-damaged kernels (FDK), can negatively impact human and livestock health (McMullen et al. 1997). Although Fusarium graminearum is the primary causal agent of FHB, several other species including F. poae could pose a risk by producing dangerous mycotoxins such as nivalenol, DON, HT-2, and T-2 (Stenglein 2009). Severe FHB epidemics on wheat have occurred in recent years along with increased corn acreage across the southeast US specifically in Georgia (Ghimire et al. 2020). Five symptomatic wheat heads displaying bleaching symptoms were randomly collected from 19 different fields across 13 counties of Georgia in late spring of 2018. Infected kernels were dipped in 6% sodium hypochlorite for 10 min and rinsed three times with sterilized water. Blot dried kernels were placed on potato dextrose agar (PDA) and incubated for 7 days at 25°C under 12-h photoperiod. Three isolates (GA18W-2.1.6, GA18W-6.1.4, and GA18W-10.2.3) from Terrell, Peach, and Sumter counties exhibited dense, whitish mycelium colony typical of F. poae (Leslie and Summerell 2006). When grown in carboxymethylcellulose broth, isolates produced globose to piriform microconidia (5.1 to 12.4 µm by 4.4 to 11.2 µm) that were aseptate or had a single septation. The morphological identification was further confirmed by DNA sequencing. Single hyphal tip isolates were grown on cellophane overlain on PDA for 10 days. Fungal DNA was extracted using a Qiagen DNeasy Plant Mini Kit. Genomic DNA was sequenced using TEF1 and TEF2 primer pairs that target the translation elongation factor 1-α (EF1-α) locus (O'Donnell et al. 1998). BLASTn query of the obtained sequences of GA18W-2.1.6 (accession no. MT856907) and GA18W-10.2.3 (accession no. MT856909) were identified as F. poae with a 99% sequence homology with GenBank reference accession MK629641, while GA18W-6.1.4 (accession no. MT856908) displayed 100% similarity with F. poae accession KJ947343. Koch's postulates were performed under greenhouse conditions. Three seeds of the FHB susceptible wheat cultivar 'SS8641' were planted in individual cone-tainers with three replications (two cone-tainers/replicate). Wheat plants were vernalized for six weeks and then moved back to the greenhouse. Each F. poae isolate was spray inoculated (50,000 spores/ml) at the flowering stage onto 18-24 wheat heads. A field isolate of F. graminearum was included as a positive control whereas heads mock-inoculated with water were used as a negative control. Inoculated wheat heads were incubated in black plastic bags for 48 hours. Disease severity and FDK were recorded three weeks post inoculation. Disease severities were 6.7% (GA18W-2.1.6), 8.3% (GA18W-10.2.3), and 15.2% (GA18W-6.1.4) compared to 90.0% in the positive control similar to Arrúa et al (2019). No symptoms were observed in the negative control. FDK was 18% (GA18W-2.1.6), 28% (GA18W-10.2.3) and 44% (GA18W-6.1.4). F. poae was re-isolated from the infected heads and found to be morphologically identical to the isolates used for inoculation. To our knowledge, this is the first report of F. poae associated with FHB of wheat in the state of Georgia, USA. F. poae isolates from Georgia might produce mycotoxins in addition to reducing grain yield which needs further study.

First report of Xanthomonas hortorum causing bacterial leaf spot of lavender (Lavandula × intermedia) in Ohio.

In July 2018, a sample of lavender var. Grosso (Lavandula × intermedia 'Grosso') from Miami County, OH was received by The Ohio State University Vegetable Pathology Laboratory in Wooster. Lavender plants were field-grown in sandy clay soil with plastic mulch under drip irrigation. Disease incidence ranged from 0 to 32% depending on variety. Leaves and stems showed dark necrotic lesions that varied from roughly circular (ca. 0.3 to 0.5 mm diameter) to large coalesced necrotic areas surrounded by a water-soaked halo. Bacterial streaming from lesions was observed microscopically. Leaf tissue pieces (~0.5 cm2) were surface sterilized in 70% ethanol for 30 seconds and rinsed in sterile deionized water. The tissue was sliced aseptically into smaller sections in 100 μl sterile water and the bacterial suspension was streaked on yeast dextrose calcium carbonate agar medium. Ten yellow Xanthomonas-like colonies were selected after 72 hours of incubation at 28ºC in the dark. Strains were gram negative, oxidase negative and caused hypersensitive reactions on Nicotiana benthamiana (L.). All strains were genotyped after whole-cell DNA extraction by BOX-PCR (Louws et al. 1999) and had the same banding profile. Four 8-wk-old lavender plants (Lavandula dentata and Lavandula × ginginsii 'Goodwin Creek Gray') were spray-inoculated with a 106 CFU/ml suspension of strain SM175-2018 in sterile water. Control plants were sprayed with sterile water. Plants were kept in plastic bags for the first 48 h at 28°C with a 14-h photoperiod. Water-soaked necrotic lesions appeared 14 days after inoculation with SM175-2018, whereas mock-inoculated plants did not show symptoms. Bacterial isolation from symptomatic leaf tissue was carried out as described above. The BOX-PCR profile of the re-isolated strain was identical to that of SM175-2018. Multilocus sequence analysis of the housekeeping genes fuyA, gyrB, and rpoD was performed (Accession numbers: MT764834 - MT764836). The resulting concatenated data set was used to perform a phylogenetic analysis using maximum likelihood criteria to evaluate relationships with closely related Xanthomonas spp. using published reference sequences (Young et al. 2008). SM175-2018 was assigned to the X. hortorum clade (Moriniere et al. 2020) with strong bootstrap support. The strain was subjected to whole genome analysis. Genomic DNA was extracted using a QIAGEN Genomic DNA buffer set with genomic-tip 100/G following manufacturer's protocol and sequenced using the iSeq-100 Illumina platform with the Nextera DNA Flex Library Prep protocol kit and Nextera DNA CD indexes. Average nucleotide identity (ANI) analysis was performed with the ANI-Matrix software Enveomics tool (Rodriguez-R and Konstantinidis 2016) using the sequenced genome (NCBI GenBank Biosample no. SAMN11831455) and those of other X. hortorum (Vauterin et al. 1995) bacteria (pvs. hederae, carotae, vitians). SM175-2018 shared a 96% ANI with other X. hortorum strains. X. hortorum is associated with bacterial leaf spot of carrot (Scott and Dung, 2020) and also reported on ornamental plants (Mirik et al. 2010, Oliver et al. 2012, Roberts and Parkinson 2014, Klass et al. 2019), however additional research is needed to establish the host specificity of lavender strains. To our knowledge this is the first report of X. hortorum causing bacterial leaf spot of lavender in Ohio. The disease may negatively impact the yield and quality of flowers used in production of lavender oils and essences.

First Report of Stem Canker of Tomato Caused by Fusarium striatum in Mexico.

Tomato (Solanum lycopersicum L.) is an important vegetable crop in Mexico. During 2015 and 2016, symptoms of stem canker were observed on tomato plants in two greenhouses located in the states of Sinaloa and San Luis Potosi, Mexico. Symptomatic plants exhibited dark brown cankers on stems and brown discoloration of the pith, as well as chlorosis, senescence of leaves, and wilting. At the base of diseased plants, orange-red perithecia were developed. Disease incidence ranged 1-5% in the two greenhouses. Pieces from symptomatic stems were surface disinfested by immersion in a 1% sodium hypochlorite solution for 2 min, rinsed in sterile distilled water, and placed in Petri plates containing acidified potato dextrose agar (APDA). The plates were incubated at 25 ºC for 6 days under a 12-h photoperiod. Fusarium-like colonies were consistently isolated and 10 monoconidial isolates were obtained. A representative isolate of each site was selected for morphological characterization, phylogenetic analysis, and pathogenicity tests. The two isolates were deposited in the Culture Collection of Phytopathogenic Fungi at the Research Center for Food and Development (accession nos. CCLF11 and CCLF12). Colonies on PDA at 25°C for 7 days exhibited moderate and cream aerial mycelium. Microscopic examination showed falciform, hyaline macroconidia (n= 100), 4- to 5-septate, measuring 40 to 75 × 4 to 6 µm. Microconidia (n= 100) were cylindrical, hyaline, 0- to 1-septate, measuring 7.8 to 9.5 × 3.1 to 4.8 µm. Chlamydospores were absent. To further identify the pathogen, total DNA was extracted, and the RNA polymerase's second largest subunit (RPB2) and a portion of the translation elongation factor 1-alpha (TEF1-α) were amplified by polymerase chain reaction (PCR) using the primers 5f2 (Liu et al. 1999)/7cr (Reeb et al. 2004) and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively. The sequences were deposited in GenBank (accession nos. RPB2: MT263727, MT263728; and TEF1-α: MT249025, MT249026). A phylogenetic analysis was performed by the Maximum Likelihood method with a combined dataset of RPB2 and TEF-1α sequences for Fusarium and Neocosmospora species (Sandoval-Denis and Crous 2018). The phylogenetic tree grouped the two isolates CCLF11 and CCLF12 within the F. striatum clade with 99% of bootstrap support. Pathogenicity of the two isolates was verified by inoculation of colonized PDA plugs (5 mm diameter) on the wounded stem surface of 10 2-month-old tomato plants from cv. Sun 6200. Ten control plants were inoculated with PDA plugs without mycelia. All plants were kept under greenhouse conditions at 25 to 35°C and regularly watered. Symptoms of stem canker were observed on all inoculated plants after 15 days, whereas stems from control plants remained healthy. After 45 days, perithecia were observed on stem cankers. Koch´s postulates were fulfilled when the fungus was re-isolated from the stems of inoculated plants and not from control plants. Fusarium striatum has been previously reported causing stem canker of tomato in greenhouses in Canada and the USA (Moine et al. 2014). To our knowledge, this is the first report of F. striatum causing stem canker of tomato in Mexico. This fungal pathogen represents a severe threat and has the potential to cause significant yield losses in tomato greenhouses, so further research is required to define effective management strategies.

Increasing Levels of Supplemental LED Light Enhances the Rate Flower Development of Greenhouse-grown Cut Gerbera but does not Affect Flower Size and Quality

To investigate the influence of supplemental lighting intensity on the production (i.e., rate of flower development, flower quality, and yield) of cut gerbera during Canada’s supplemental lighting season (November to March), trials were carried out at a research greenhouse. Five supplemental light emitting diode (LED) light intensity (LI) treatments provided canopy-level photosynthetic photon flux densities (PPFD) ranging from 41 to 180 µmol m−2 s−1. With a 12-h photoperiod, the treatments provided 1.76 to 7.72 mol m−2 d−1 of supplemental light. Two cultivars of cut gerbera (Gerbera jamesonii H. Bolus ex Hook.f) were used to evaluate vegetative growth and flower production. Plugs of ‘Ultima’ were assessed for vegetative growth and rate of flower development. There were minor LI treatment effects on number of leaves and chlorophyll content index and flowers from plants under the highest versus the lowest LI matured 10% faster. Reproductively mature ‘Panama’ plants were assessed for flower yield and quality. ‘Panama’ flowers from the highest LI treatment had shorter stems than the three lowest LI treatments, and flowers from the middle LI treatment had larger diameter than the other treatments. Flowers from the lowest LI treatment had lower fresh mass than the three highest LI treatments. There were linear relationships between LI and numbers of flowers harvested, with the highest LI treatment producing 10.3 and 7.0 more total and marketable flowers per plant than the lowest LI treatment. In general, increasing levels of supplemental light had only minor effects on vegetative growth (young plants) and size and quality of harvested flowers (mature plants), but flowers from plants grown under higher LIs were more numerous and matured faster.

Sanitary quality of seed of Paspalum species

Abstract Brazil has been developing research for selection of native species of Paspalum for use as turfgrass, nevertheless little is known about the sanitary quality and pathogens associated to the seeds, being the aims of this study. Subsamples of 100 seeds of two accessions of Paspalum notatum Flügge (PN 01 and PN 04) and one accesses of Paspalum lepton Schult. (PL 01) were submitted to the blotter test at 20 °C with a 12-h photoperiod for 7 days. The Penicillium and Curvularia genus incidence were highest in the three accessions of Paspalum. The seed health quality was variable between accessions, with more than 56% of incidence of the fungus Curvularia on the seeds. This study reported for the first time the occurrence of Curvularia spp. and Cladosporium herbarium in seeds of P. lepton, in the Northeast region of Brazil.

Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose.

Despite the increasing use of protoplasts in plant biotechnology research, shoot regeneration from protoplasts remains challenging. In this study, we investigated the factors involved in protoplast isolation, callus induction, and shoot regeneration in Petunia hybrida cv. Mirage Rose. The following conditions were found to be most optimal for protoplast yield and viability: 0.6 M mannitol, 2.0% cellulase, and 6 h digestion time. A plating density of 10 × 104 protoplasts/mL under osmoticum condition (0.58 M mannitol) showed high microcolony viability in liquid culture. The Kao and Michayluk medium was found to be appropriate for callus proliferation from microcalli under a 16-h light photoperiod. Calli cultured in Murashige and Skoog medium containing 1.0 mg/L 6-benzylaminopurine and 0.2 mg/L 3-indole butyric acid showed the highest shoot regeneration frequency and number of shoots obtained per explant. Random amplification of polymorphic DNA analysis showed that the protoplast-derived shoots exhibited the same banding patterns as those of donor plants. Collectively, these findings can contribute to solving problems encountered in protoplast isolation and shoot regeneration in other petunia cultivars and related species. As the protocol developed by us is highly reproducible, it can be applied in biotechnology research on P. hybrida cv. Mirage Rose.

First Report of Mycoleptodiscus terrestris Causing Crown and Root Rot of Alfalfa (Medicago sativa) in Minnesota.

Mycoleptodiscus terrestris (Gerd.) Ostaz. has been studied extensively as a potential mycoherbicide against aquatic weeds since the early 1970s (Mathur and Gehlot 2018; Shearer 1998). However, it is also a pathogen on many legumes including alfalfa (Gerdemann 1953; Smith et al. 1998). In August 2019, alfalfa plants with stunted yellow foliage and rotted stems at the base of the crown were observed in a variety trial planted in spring 2018 at the University of Minnesota, St. Paul, Minnesota. Affected plants were in patches across the field with approximately 20% of plants showing symptoms. Plants from patches had few lateral and fibrous roots and dark lesions appeared in the crown and root tissue. Tissue samples (5 mm2) from 10 plants with symptomatic crown and root tissues were surface disinfested with 70% ethanol for 5 min, followed by 30 s in 10% NaOCl, and then rinsed three times in sterile distilled water. Samples were air-dried and placed on water agar (WA) amended with 25 µg/ml rifampicin. Plates were incubated at room temperature for 3 days and fungal hyphal tips from tissue samples were transferred to potato dextrose agar (PDA). After 7 days, isolates were tentatively identified by morphological characteristics as M. terrestris (Ostazeski 1967). The hyaline mycelia turned from olive-gray to dark gray with age. Abundant dark microsclerotia formed 5 days after incubation that varied in size and shape, measuring 300-860 x 270-600 µm. Molecular identification of two representative isolates (DAZ5 and DAZ9) was done by sequencing the internal transcribed spacer (ITS) regions of the rDNA, the translation elongation factor 1α (TEF1), and the second largest subunit of RNA polymerase II (RPB2) genes. For both isolates the ITS, TEF1, and RPB2 genes were amplified and sequenced with primers ITS1/4 (White et al. 1990), 983F/1567R (Rehner and Buckley 2005), and fRPB2-7cR/RPB-5F2 (Liu et al. 1999; Sung et al. 2007), respectively. ITS amplification conditions followed White et al. (1990), the TEF and the partial RPB2 gene was obtained by using a touchdown PCR protocol as described in Rehner and Buckley (2005) and Woudenberg et al. (2017), respectively. Based on a BLAST search of the NCBI nucleotide database, the ITS sequences (GenBank MN851265.1 and MN851266.1) had 100% identity with M. terrestris strain CBS 231.53 (MK487754.1). The TEF1 (MN873019, MN873020) and RPB2 sequences (MN873021, MN873022) had 100% identity with M. terrestris strain IMI 159038 (MK495977.1, MK492735.1). A pathogenicity test was performed by inoculating five 3-week-old alfalfa plants per cultivar (cv. DKA44-16RR, Vernal, 53V52, and Agate) with isolates DAZ5 and DAZ9. Plants were inoculated around the exposed stem base with three 5 mm diameter PDA plugs from a culture of M. terrestris covered with microsclerotia. Control plants were inoculated with sterile PDA plugs. After inoculation plants were incubated at 25 0C with a 16-h photoperiod in a growth chamber. The first symptoms appeared two months after inoculation on all cultivars as a dark lesion at the stem base followed by yellowing and rotting of stems. Inoculated plants had more fibrous roots than the controls. The control plants were symptomless with healthy root development. The pathogen was re-isolated from all infected alfalfa cultivars and had the same morphology as isolates DAZ5 and DAZ9, fulfilling Koch's postulates. To our knowledge, this is the first report of M. terrestris causing crown and root rot disease of alfalfa in Minnesota.

First Report of Anthracnose Caused by Colletotrichum karstii on Fatsia japonica in Sichuan, China.

Fatsia japonica (Thunb.) Decne. & Planch. (Araliaceae), an evergreen shrub, is widely planted in urban settings at the south of the Yangtze river in China. Leaf-spot symptoms were observed on 120/200 F. japonica plants at Sichuan Agricultural University. Initially, yellowish spots appeared on the leaves and became white with age. The spots continued to expand with time developing irregular margins eventually encompassing the entire leaf. Eventually, diseased leaves became curled and died. Four single-spore isolates were obtained following Chomnunti et al. (2014). The colonies developed on potato dextrose agar (PDA) were white to grey with fluffy aerial hyphae. The base of the mycelium was yellow. On the host conidiomata circular to elliptical, measured 108-335 × 107-250 μm (n=20). Conidiophores were bell-shaped to cylindrical, hyaline, usually 1-celled, unbranched, 9.7-23.6(-28) × 3.5-5.5 μm (x̅ =16.5 × 4.6 μm, n=15). Conidia measured 12.5-17.6 × 5.1-7.9 μm (x̅ =14.9 × 6.7 μm, n=25), were hyaline, straight, unicellular, cylindrical. Appressoria formed abundantly on slides, measured 12.4-25.4(-29.6) × 6.3-18.4 μm (x̅ =18.9 × 13.4 μm, n=30), were dark brown to black, elliptical to irregular. Ascomata on PDA were brown to dark brown, global, covered by aerial mycelium, and setae were absent. Ascospores measured 13.2-19.1 × 4.0-6.1 μm (x̅ =16.0 × 4.8 μm, n=20), and one-celled, hyaline, slightly curved. The representative isolate SICAUCC 20-0010 was used for Genomic DNA extraction following the instructions of Plant Genomic DNA extraction kitTM (Tiangen, China), and the internal transcribed spacer (ITS, MT393946), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, MT425977), beta-tubulin (TUB2, MT425978), chitin synthase (CHS-1, MT425976) and actin (ACT, MT425975) genes were amplified with ITS5/ITS4 (White et al. 1990), GDF/GDR (Guerber et al. 2003), T1/Bt2b (O'Donnell et al. 1997), CHS-79F/CHS-354R and ACT-512F/ACT-783R (Carbone et al. 1999) primers, respectively. ITS, GAPDH, CHS-1 and ACT blast showed 100% homology with sequences of Colletotrichum karstii (Yang et al. 2011, Damm et al. 2012, Xu et al. 2019), viz. JQ005186 (555/555 bp), MK359214 (238/238 bp), JQ005360 (274/274 bp), MK359211 (269/269 bp), respectively. TUB2 showed 99.60% homology with sequences JQ005620 (495/497 bp). The fungus was identified as C. karstii based on morphology and a multigene phylogeny (Fig. 1). Four leaves from each of three healthy F. japonica plants were sprayed with a conidial suspension (1 × 105 conidia/ml) of the isolate SICAUCC 20-0010. Two additional mock-inoculated control plants were sprayed with sterile distilled water. Plants were incubated in a greenhouse at 26℃ and 90% relative humidity with a 12-h photoperiod. The leaves developed chlorotic lesions 6 days post-inoculation and by twenty-two days post-inoculation the remaining signs and symptoms had developed. No symptoms developed on the mock-inoculated controls. The fungus isolated from inoculated plants was morphologically identical to the original pathogen, fulfilling Koch's postulates. The anthracnose on F. japonica caused by C. fructicola and C. gloeosporioide have been reported in China (Wang 2007, Shi et al. 2017). However, this is the first report of anthracnose caused by C. karstii on F. japonica. Field observations indicated that this disease mainly occurs on old and weakened leaves. This disease affects the aesthetic appearance of the plants reducing their appeal as landscape plants.

The effect of supplementary ultraviolet wavelengths on the performance of broiler chickens

Qualities of the light environment, such as the spectral composition of light, have been shown to impact growth and performance of broiler chickens. UVA light is visible to broiler chickens, whereas UVB wavelengths promote endogenous vitamin D synthesis, which could support their rapid development. The aim of the current study was to investigate the impacts of supplementary UVA and UVB wavelengths on performance indicators of broiler chickens.Day-old Ross 308 chicks (n = 638), reared to a target stocking density of 33 kg/m2, were randomly assigned to 1 of 3 lighting treatments: A) White light emitting diode (LED) and supplementary UVA LED lighting (18-h photoperiod); B) White LED with supplementary UVA and UVB fluorescent lighting providing 30 μW/cm2 UVB at bird level (lights on for 8 h of the total photoperiod to avoid overexposure of UVB); and C) White LED control group, representative of farm conditions (18-h photoperiod).Mortality was recorded, and broiler chickens were individually weighed at 8, 15, 22, 27, and 34 D of age. Generalized linear models and nonlinear mixed effects models (Gompertz curve) were fitted to determine the effects of UV wavelengths on broiler mortality and growth performance.UV did not impact breast or leg weight of broiler chickens but was associated with differences in mortality, growth, and end weight. Broiler chickens provided with UVA for the full 18-h photoperiod had slower initial growth than control broilers and a reduction in mortality. Results from male broilers reared with supplementary UVA + UVB for 8 h indicated they could reach finishing weights sooner than controls, which supports the potential for UVA + B to improve the growth performance of males.Results suggest that the provision of supplementary UVA + UVB wavelengths may improve the performance of male broiler chickens. The reduction in mortality in the UVA only treatment may warrant further investigation. The inclusion of UV wavelengths within poultry lighting regimes represents a promising area of further study.

First Report of Sclerotinia sclerotiorum Causing Strawberry Fruit Rot in Florida.

During the 2019-2020 Florida strawberry season (October to April), a strawberry (Fragaria × ananassa) fruit rot was observed in two fields (Plant City and Wimauma, FL) with up to 5% incidence on short-day cultivars SensationTM Florida127, and Florida Brilliance. Symptoms on pink and ripe fruit consisted of softening, discoloration, watery rot with white fuzzy mycelium, and initial sclerotium formation. Diseased tissue was placed on General Isolation (GI) medium (Amiri et al. 2018) and incubated at 25°C under a 12-h photoperiod. A fungus producing spreading cottony white colonies with dark sclerotia near the outer edges of the plates was consistently isolated. One isolate from each cultivar field (20-51 and 20-55) was selected for identification and pathogenicity tests. Apothecial production was induced following the protocol of Li and Rollins (2009), and apothecia (n=30) had an average diameter of 4.5 (3.5 to 7.2) mm. Sclerotia were 3.6 (2.5 to 6.2) mm by 4.5 (3.1 to 5.9) mm (n=30) in size. Dimensions of asci were 130.2 (115.1 to 160.5) μm by 8.5 (6.2 to 13.1) μm (n=30), and those of ascospores were 12.2 (10.8 to 14.6) μm by 6.8 (5.7 to 8.1) μm (n=30). Based on these morphological features, the pathogen was tentatively identified as Sclerotinia sclerotiorum (Lib.) de Bary (Maas 1998). DNA was extracted from the same two isolates using the FastDNA kit (MP Biomedicals, Solon, OH), and the ribosomal internal transcribed spacer (ITS) region was amplified using the primers ITS1 and ITS4 (White et al. 1990). Sequences were deposited in GenBank (accession nos. MT378215 and MT378216). BLASTn searches revealed that isolates 20-51 and 20-55 were 99.62% identical (526 / 528 bp) to S. sclerotiorum CBS 499.50 (MH856725.1). Immature pink fruit of SensationTM 'Florida127' were harvested, surface disinfested in bleach solution (0.08% NaClO) for 90 sec, rinsed twice with deionized water, then placed into styrofoam egg cartons inside clean plastic boxes (30x24x7 cm) containing 150 ml of sterile deionized water to maintain moisture, and kept at 25°C with a 12-h photoperiod. Ascospores and sclerotia were used for inoculation tests with three repetitions in an egg carton containing 12 fruit each per isolate and inoculation method. The experiment was repeated once. Fruit were inoculated by placing 20 μl of a 1 × 106 ascospores/mL suspension or a single sclerotium on the upper half of the fruit. Controls were included, by placing 20 μl of sterile DI water or fruit with no sclerotia. Evaluations were done 6, 10, and 15 days after inoculation (DAI). Control fruit remained healthy, while inoculated fruit developed symptoms of softening and discoloration. For ascospore inoculation, disease incidence was 55 (± 5) and 78% (± 4), for 6 and 15 DAI, respectively, whereas for sclerotia inoculation incidence was 100% 6 DAI. Morphologically identical fungi to the original isolates were re-isolated from the diseased fruit. No other fruit decay fungi were observed. S. sclerotiorum has been previously reported causing strawberry fruit rot in Washington state in the United States, England, Israel, and Scotland (Alcorn 1966; Maas 1998; McLean 1957). It has also been listed in the indices of plant diseases from Florida, North Carolina, and California as causing crown rot (Farr and Rossman 2020). To our knowledge, this is the first report of S. sclerotiorum causing strawberry fruit rot in Florida. The pathogen is an aggressive necrotroph with prolonged survival and affects several vegetable crops grown in Florida (Paret et al. 2018). Because only the strawberry beds are fumigated, sclerotia remaining in the alleys could serve as inoculum sources. Currently, the disease is rare and of minor significance to strawberry production. However, efforts should be implemented to monitor its occurrence and spread.

First Report of Leaf Blight Caused by Cladosporium perangustum on Livistona chinensis in China.

Livistona chinensis (Jacq.) R.Br. ex Mart. belonging to monocotyledonous Palmaceae, is widely distributed in Eastern Asia, and a common ornamental plant in Southern China (Li et al. 2019, Wu et al. 2019). In November 2019, epidemics of leaf blight were discovered on aged leaves of L. chinensis in the campus of Sichuan Agricultural University, where disease incidence was up to 90% among the cultivated plants, but without resulting in the death of the host. Initial symptoms appeared chlorotic spots on the upper part of leaves, then spots expanded and turned brown. Subsequently, the enlarged spots developed necrotic tissues in the center with brown to dark brown margin. At later stages, conidia and conidiophores were observed on the surface of lesions with the aid of a microscope. As the disease progressed, multiple lesions usually coalesced to cause extensive tissue necrosis at third or more parts of the leaf. Three isolates were obtained from diseased leaves with a single spore isolation referred to Chomnunti et al. (2014), and cultured on potato dextrose agar (PDA) under a 12-h photoperiod and 25±1℃. The colonies were gray to gray-olivaceous with an olivaceous-black reverse side, and measured approximately 7 cm in diameter after 6 days. The conidiophores were macronematous, semimacronematous or micronematous, erect, straight or slightly flexuous, filiform to narrowly cylindrical-oblong, occasionally branched, pale olivaceous-brown, wider at the base, 1 to 8 septate, 14-104 × 3-6.8 µm (x̅ =57.7 × 4.9 µm, n=30). Conidiogenous cells were integrated, terminal, intercalary or pleurogenous, cylindrical-oblong, 9-21 × 4-5.9 µm (x̅ =15 × 4.9 µm, n=20), thick walled, usually aseptate, occasionally 1-2-septate, with 1-5 apically crowed loci. Ramoconidia were almost oblong, aseptate, 4-8.4 × 3-5.7 µm (x̅ =6.9 × 4.6 µm, n=20). Conidia were numerous, catenate, in branched chains, small terminal conidia, subglobose or ovoid, and intercalary conidia were ovoid, limoniform, fusiform, aseptate, 2.9-5.5 × 2-5 µm (x̅ =4 × 3.5 µm, n=20). The representative isolate SICAUCC 20-0007 was used for Genomic DNA extraction, and the internal transcribed spacer (ITS), translation elongation factor 1 alpha (TEF 1-α) and partial actin (ACT) regions were amplified using the primer pairs ITS5/ITS4 (White et al. 1990), EF1-728F/EF-2 (O'Donnell et al. 1998, Carbone et al. 1999) and ACT-512F/ACT-783R (Carbone et al. 1999), respectively. Nucleotide BLAST in GenBank revealed identity with those of C. perangustum, the ITS (MT427730) had 100% identity with MF473185 and MF473177, TEF 1-α (MT441922) had 100% identity with HM148379 and MF473595, ACT (MT441917) had 100% identity with HK521577 and MH047335. The phylogenetic tree combined with ACT, ITS, and TEF 1-α genes (Fig. 1) and morphological characteristics confirmed the identification as C. perangustum defined by Bensch et al. (2010). To determine pathogenicity, five potted plants were inoculated with conidial suspensions (105 conidia/ml) prepared from 1-week-old cultures of the isolate SICAUCC 20-0007 onto the wounded sites via pin-prick inoculation (five to eight leaves per plant with roughly 1 to 2-year-old), and the same amount of healthy plant was sprayed with distilled water as controls. Plants were sprayed regularly with distilled water every day and placed under a 12-h photoperiod and 25±1℃. About fifteen days later, faint yellow to yellowish-brown spots were found on inoculated leaves and were similar to those previously observed and no symptoms developed on the control plants. After a month, leaf blight was observed and the pathogenic fungus was re-isolated from the inoculated tissues. Previous reports have shown that C. perangustum causes leaf spot on Myrica rubra (Lour.) Siebold & Zucc. in China (Lu et al. 2015). To our knowledge, this is the first report of C. perangustum causing leaf blight of L. chinensis in the world. This disease potentially reduces the ornamental value under favorable conditions, and proper control strategies should be implemented.

First report of sour rot of strawberry caused by Geotrichum candidum in the United States.

Strawberry (Fragaria x ananassa) is an important crop in the U.S., and Florida is the second major producer in the country. In January 2020, an unknown fruit rot was observed in two strawberry fields in Dover (seedling selection) and Plant City (cultivar Florida127), Florida. Disease incidence varied from less than 1% in one field to up to 15% in the second field during some harvests where over-ripe fruit were present. Affected fruit had a water-soaked soft rot with a sour smell, and sometimes with white mycelium on the fruit surface. Direct isolation was performed from symptomatic fruit from each area by touching the surface of a lesion with a sterile needle and streaking the fungus over general isolation medium (Amiri et al. 2018). The fungus was incubated at 25°C and 12-h photoperiod for five days. Four single-spore isolates (20-46 and 20-47 from Plant City; 20-49 and 20-50 from Dover) obtained from different colonies were grown on potato dextrose agar (PDA). Colonies were white to cream, flat, with a powdery surface, and had a characteristic sour odor. Hyphae were hyaline, septate, growing in a dichotomous pattern and often disarticulating into arthroconidia, which were unicellular, spherical to cylindrical, measuring 4.8 to 9.5 × 3.5 to 5.6 µm (n = 50). Based on the morphological characteristics, the fungus was identified as Geotrichum candidum (De Hoog et al. 1986). To confirm the identity of the isolates, the ribosomal internal transcribed spacer (ITS-rDNA) and the translation elongation factor 1-alpha (TEF1-α) gene regions were amplified and sequenced using the primers ITS1/ITS4 (White et al. 1990), and EF1-728F/EF1-1567R (Carbone and Kohn 1999), respectively. Consensus sequences were deposited in the GenBank (accession numbers MT353975 to MT353978 for ITS-rDNA, and MT346367 to MT346370 for TEF1-α). The BLASTn analysis revealed 99% identity with reference sequences of G. candidum for ITS-rDNA (KU373122) and TEF1-α (MK397513). Phylogenetic analysis, including ITS sequences of G. candidum and other spp., obtained from GenBank, was performed using maximum likelihood and Bayesian inference methods, implemented in MEGA-X and MrBayes, respectively. The isolates were grouped within the G. candidum clade. The pathogenicity of the fungus (isolate 20-46) was evaluated in ripe and green strawberry fruit (cultivar Florida127). Non-wounded and superficially wounded (with a sterilized needle) fruit were inoculated with 20 µl of a spore suspension (1 × 107 spore/ml), prepared by washing the surface of a 5-day-old colony on PDA, with sterile deionized water (SDW) plus 0.01% (v/v) Tween 20. Wounded and non-wounded control fruit were treated with SDW. Fruit were maintained in moist containers at 25°C. Each treatment consisted of 12 fruit (three replicates, each containing four fruit) and was evaluated daily for 10 days. Symptoms of sour rot were visible as soon as 48 hr after inoculation, but only on ripe (100% incidence) and green (58% incidence) wounded fruit. Symptoms progressed to an intense water-soaking with the presence of a typical white mycelium on the surface. Control fruit remained symptomless. The fungus was successfully recovered from symptomatic fruit, fulfilling Koch's postulates. Although strawberry fruit rots caused by G. candidum have already been reported in Pakistan and China (Hussain et al. 2016; Ma et al. 2018), this is the first report of this species causing sour rot on strawberry in the U.S. The disease may be a minor problem on damaged or over-ripe fruit, but further studies might be needed to determine its importance, distribution, and potential strategies for control.

Blue LED light promotes indican accumulation and flowering in indigo plant, Polygonum tinctorium

The photoperiod is a day-length-dependent seasonal change of physiological or developmental activities, such as flowering, in many plant species. Polygonum tinctorium (P. tinctorium) is an important industrial crop producing indigo blue dyes, and indican is an important substance as a precursor of indigo. Here, we report the day-length-dependent responses of growth, flowering, and indican synthesis in P. tinctorium. Indigo plants were grown in a hydroponic system under artificial light conditions in a completely-controlled plant factory. The growth parameters and indican content of leaves were measured and compared. Certain growth parameters (such as fresh weight and number of leaves) under 24-h continuous irradiation were significantly higher than those under other day-length conditions. Under 12-h photoperiod conditions, the flowering rate of plants with blue LED treatment increased six-fold compared with fluorescent white light treatment, while none of the plants flowered with red LED treatment. In the leaves, the relative expression levels of Pt’IGS and Pt’BGL were significantly higher in newer leaves compared to older ones. Indican content was greatly enhanced by blue light under 24-h continuous irradiation, which was reflected by increased expression levels of Pt’IGS. These findings demonstrate that there is a new regulatory mechanism for the indican synthesis pathway through blue light signalling. Blue light provides feasible strategy for artificially regulating indican synthesis and flowering in P. tinctorium.

Polyphenolic Composition and Allelopathic Potential of Senna cearensis Afr. Fern. (Fabaceae)

Chemical compounds from the secondary metabolism of a plant, termed as allelochemicals, may interfere with the germination and/or development of other plant species, a phenomenon known as allelopathy. The aim of this study was to analyze the allelopathic potential of the Senna cearensis Afr. Fern. aqueous extract at various concentrations, collected from an area in the Chapada do Araripe during two different seasons (dry and rainy), on Lactuca sativa L. and Cenchrus echinatus L. seeds and seedlings, in addition to analysing the extract’s phytochemical prospection. The bioassays were conducted in a germination chamber at 25 °C with a 12h photoperiod and consisted of four treatments (extract at 12.5%, 25%, 50% and 100%) and a control group (distilled water). The variables analyzed were: seed germination, germination speed index, radicle length and caulicle seedling length. The means from each variable were submitted to an analysis of variance and compared using Tukey’s test (p<0.05), through the ASSISTAT. Quantitative prospection of the extract was performed using HPLC-DAD. The extracts reduced L. sativa seed germination, however it did not affect C. echinatus seeds. The extracts caused a reduction in germination rate and presented inhibitory effects on seedling development for both receptor species. Quercitrin was the major compound in both extracts. The allelopathic activity of the S. cearensis aqueous leaf extract collected during dry or rainy periods may be a viable alternative for weed control. Further studies addressing fractionation for separation and the possible isolation of substances responsible for the observed allelopathic activity are necessary.

Optimizing mass production of Trichoderma asperelloides by submerged liquid fermentation and its antagonism against Sclerotinia sclerotiorum.

Commercial products based on Trichoderma are obtained mainly from solid-state fermentation. Submerged liquid fermentation is the most appropriate method compared to the solid medium for large-scale production of Trichoderma spp. The present study aimed to optimize the combination of key variables that influence the liquid fermentation process of Trichoderma asperelloides LQC-96 for conidial production coupled with its efficiency in the control of Sclerotinia sclerotiorum. In addition, we verified whether the optimized culture conditions can be used for the conidial production of Trichoderma erinaceum T-12 and T-18 and Trichoderma harzianum T-15. Fermentation studies were performed in shake flasks following a planned experimental design to reduce the number of tests and consumable costs. The effect of temperature, pH, photoperiod, carbon:nitrogen ratio and water activity on conidial production were assessed, which of pH was the only meaningful factor contributing to increased conidial production of T. asperelloides LQC-96. From the five variables studied initially, pH and C:N ratio were further used in the second design (rotational central composite design-RCCD). Hence, the best conditions for the production of T. asperelloides LQC-96 conidia by liquid fermentation consisted of initial pH of 3.5, C:N ratio of 200:1 at 30°C, without glycerol, and under 24h photoperiod. The highest conidial concentration was observed after seven days of fermentation. Under these optimal conditions, T. erinaceum T-12 and T-18, and T. harzianum T-15 were also cultivated, but only LQC-96 efficiently parasitized S. sclerotiorum, precluding sclerotium myceliogenic germination. Our findings propose optimal fermentation conditions that maximize conidial production of T. asperelloides as a potential biofungicide against S. sclerotiorum.

The establishment of callus and cell suspension cultures of Hylocereus costaricensis for the production of betalain pigments with antioxidant potential

Hylocereus costaricensis, commonly known as red-purple dragon fruit or pitaya, is a climbing vine cactus that is currently widely grown as a fruit crop and has gained popularity as a potential source of both natural colourants and antioxidants contributed by betalain compounds. Betalains comprised of red-purple betacyanins and yellow betaxanthins. This study aimed to establish callus and cell suspension cultures of H. costaricensis from in vitro germinated seedlings and to evaluate their betalain content and antioxidant profiles. Epicotyl and cotyledon segments of 60-day-old axenic seedlings were examined for their callus induction potential on half-strength Murashige and Skoog (MS) medium supplemented with various concentrations of naphthaleneacetic acid (NAA) and Thidiazuron (TDZ) under different illumination (16-h photoperiod and darkness). Different combinations of picloram and 2,4-dichlorophenoxyacetic acid (2,4-D) were subsequently tested for the induction of friable callus. The betalains, total phenolic content, total flavonoid content and antioxidant activity DFRSA in cell suspension cultures were quantified using spectrophotometric analysis. Generally, cotyledon explants cultured under 16-h photoperiod was optimum for callus induction. Compact callus with roots was formed on medium supplemented with NAA while granular callus was formed in TDZ supplemented medium. Among the 36 treatments tested, supplementation of 10 μM picloram induced friable callus with average fresh mass of 0.817 g (10.2-fold as compared to the control treatment). Picloram at 10 μM concentration was also found to be effective for callus proliferation and initiation of cell suspension culture. Yellow and red callus were produced and proliferated on the same medium through repetitive clonal selection. The higher concentration of sucrose was more effective for cell mass accumulation while lower sucrose concentration stimulated the accumulation of betalain compounds and antioxidants in the cell suspension cultures of H. costaricensis. This study indicates that callus and cell suspension culture could be used as an efficient system for sustainable production of betalain compounds which can contribute to functional food colouring and pharmaceutical industries.