First Report of Fusarium avenaceum Causing Canker Disease on Kiwi Tree in China

In May 2021, canker symptoms were detected on ‘Xuxiang’ kiwi trees in southwestern Shaanxi (Hanzhong municipality; 107.27° E, 33.23° N) in China. Seven-year-old trees exhibited black necrotic lesions and cracked areas in the trunk (Figure 1). The symptoms were observed in approximately 10% of the trees in 6 orchards (31 ha in total). Application of commercial fungicides did not control the advancement of the pathogen, and infected trees were removed to control the spread. Three samples, approximately 1 cm2 in size, of symptomatic tissue were collected and surface sterilized in 2% NaOCl for 1 min, and washed with sterile ddH2O. Four isolates showing white mycelium with yellow pigmentation were obtained after 4 days of incubation on PDA, containing chloramphenicol (50 µg/mL), at 28 ºC. The pathogen was isolated from all collected samples. ITS, EF1-α, TUB2, RPB1 and RPB2 genes were amplified using ITS1/ITS4, EF1-728F/EF1-986R, T1/T22, RPB1-5F/RPB1-8R and RPB2-5F/RPB2-7cR (strain NJC06), or RPB2-c7F/RPB2-11aR (strains NJC07 and NJC08), primers, respectively. Two isolates shared the same sequences (strain NJC08). Obtained sequences were submitted to GenBank under accession numbers MZ669205 and OL347898-OL347899 (ITS), OL439731-OL439733 (EF1-α), OL439734-OL439736 (TUB2), OL439737-OL439739 (RPB1), and OL439740-OL439742 (RPB2). The sequences shared >99% (ITS; F. avenaceum CBS 128538, MH864972), >99% (EF1-α; F. avenaceum 55-2, MN473124), 100% (TUB2; F. avenaceum SICAUCC 18-0001, MK253102), >98% (RPB1; F. avenaceum NRRL 26911, MG282372), and >98% (RPB2; F. avenaceum SICAUCC 18-0001, MK396098; or F. avenaceum FRC R-09495, CQ915486) homology to multiple F. avenaceum strains. Molecular phylogenetic tree (Figure 2) was constructed using MEGA7 with Fusarium strains found causing rot in various hosts (Wang et al. 2015), and other fungal species, such as Cadophora nalorum, Diaporthe ambigua, D. australafricana, and Neofusicoccum parvum, which were reported to cause cordon dieback on kiwi tree in Chile (Diaz et al. 2021). Microscope observations after cultivation of all isolates on barley-honey-tryptone medium (Song et al. 2020) showed the presence of septate mycelium, fusiform microconidia (8-15 µm in length, containing between 0 and 3 septa; n = 77) and chlamydospores (n = 21), and agree with the morphology of F. avenaceum (Zhao et al. 2020). To confirm pathogenicity, a sterilized spatula was used to make wounds (3 mm diameter, 1 mm depth) on the trunk of 3-months-old ‘Xuxiang’ kiwi trees. Solutions containing 1 × 106 spores/mL (20 µL) of the isolates were injected in the wounds. Sterile ddH2O was used for the control experiment. Inoculated plants were maintained in a growth chamber at 28 °C and 80% relative humidity for 4 days. The pathogen was recovered from the canker lesions, which were similar to those observed in the orchards, and its identity was confirmed by sequence analysis. The pathogen only infected wounded trees, and probably invaded the orchards during the pruning in February 2021. F. avenaceum was reported to cause canker on almond tree (Stack et al. 2020), stem rot on Anthoxanthum aristatum and Polygonatum cyrtonema (Pieczul et al. 2018; Xu et al. 2019), and root rot on carrot, Coptis chinensis and wheat (Le Moullec-Rieu et al. 2020; Mei et al. 2020; Ozer et al. 2020). Recently, F. avenaceum was found causing fruit blotch in kiwi fruit in Anhui (China) (Zhao et al. 2020). Here, F. avenaceum was found causing canker disease in kiwi tree, demonstrating the host and tissue promiscuity of this pathogen. Kiwi is an important crop in China with nearly 1.5 million tons produced in 2019. This report will help to better understand the pathogens reducing kiwi production in China.

Genome-Wide Identification and Comparative Profiling of MicroRNAs Reveal Flavonoid Biosynthesis in Two Contrasting Flower Color Cultivars of Tree Peony

MicroRNA (miRNA)-mediated gene regulation is involved in various physiological processes in plants. Flower color is one of the vital ornamental traits of tree peony (Paeonia suffruticosa Andr.). However, the yellow-flowered tree peony cultivars are particularly rare. To elucidate the miRNA-mediated gene regulatory mechanism underlying yellow pigmentation in tree peony, we combined pigment assessment, miRNA identification, expression analysis, and gene functional verification in two contrasting flower color cultivars “High Noon” and “Roufurong.” Flavones/flavonols and anthocyanins were found to be the main contributors to the coloration of “High Noon” and “Roufurong” petals, respectively. Subsequently, miRNA analysis based on available genome data identified 9 differentially expressed miRNAs and 12 relevant target genes implicated in flavonoid biosynthesis. Their dynamic expression patterns determined the key role of mdm-miR156b-PsSPL2 module in yellow pigmentation of tree peony flowers. The sequence analysis and subcellular localization validated that PsSPL2 might function as a nuclear-localized transcription factor. Overexpression of PsSPL2 in tobacco resulted in a decrease of anthocyanin content and down-regulation of NtF3′H and NtDFR transcripts. PsSPL2-silenced petals exhibited lighter yellow color, and the contents of THC, Ap, and Ch decreased significantly. Meanwhile, expression levels of PsCHS, PsCHI, and PsF3H were significantly decreased in the petals with PsSPL2 silencing, while those of PsF3′H and PsDFR were remarkably increased. This study offers a novel insight into yellow pigmentation-related miRNA regulation network in tree peony, and further provides the valuable information on physiological changes during yellow coloring process of tree peony.

From the formation of embryonic appendages to the color of wings: Conserved and novel roles of aristaless1 in butterfly development

Highly diverse butterfly wing patterns have emerged as a powerful system for understanding the genetic basis of phenotypic variation. While the genetic basis of this pattern variation is being clarified, the precise developmental pathways linking genotype to phenotype are not well understood. The gene aristaless, which plays a role in appendage patterning and extension, has been duplicated in Lepidoptera. One copy, aristaless1, has been shown to control a white/yellow color switch in the butterfly Heliconius cydno, suggesting a novel function associated with color patterning and pigmentation. Here we investigate the developmental basis of al1 in embryos, larvae and pupae using new antibodies, CRISPR/Cas9, RNAi, qPCR assays of downstream targets and pharmacological manipulation of an upstream activator. We find that Al1 is expressed at the distal tips of developing embryonic appendages consistent with its ancestral role. In developing wings, we observe Al1 accumulation within developing scale cells of white H. cydno during early pupation while yellow scale cells exhibit little Al1 at this timepoint. Reduced Al1 expression is also associated with yellow scale development in al1 knockouts and knockdowns. We also find that Al1 expression appears to downregulate the enzyme Cinnabar and other genes that synthesize and transport the yellow pigment, 3–Hydroxykynurenine (3-OHK). Finally, we provide evidence that Al1 activation is under the control of Wnt signaling. We propose a model in which high levels of Al1 during early pupation, which are mediated by Wnt, are important for melanic pigmentation and specifying white portions of the wing while reduced levels of Al1 during early pupation promote upregulation of proteins needed to move and synthesize 3-OHK, promoting yellow pigmentation. In addition, we discuss how the ancestral role of aristaless in appendage extension may be relevant in understanding the cellular mechanism behind color patterning in the context of the heterochrony hypothesis.

Integrating full-length transcriptomics and metabolomics reveals the regulatory mechanisms underlying yellow pigmentation in tree peony (Paeonia suffruticosa Andr.) flowers

Tree peony (Paeonia suffruticosa Andr.) is a popular ornamental plant in China due to its showy and colorful flowers. However, yellow-colored flowers are rare in both wild species and domesticated cultivars. The molecular mechanisms underlying yellow pigmentation remain poorly understood. Here, petal tissues of two tree peony cultivars, “High Noon” (yellow flowers) and “Roufurong” (purple–red flowers), were sampled at five developmental stages (S1–S5) from early flower buds to full blooms. Five petal color indices (brightness, redness, yellowness, chroma, and hue angle) and the contents of ten different flavonoids were determined. Compared to “Roufurong,” which accumulated abundant anthocyanins at S3–S5, the yellow-colored “High Noon” displayed relatively higher contents of tetrahydroxychalcone (THC), flavones, and flavonols but no anthocyanin production. The contents of THC, flavones, and flavonols in “High Noon” peaked at S3 and dropped gradually as the flower bloomed, consistent with the color index patterns. Furthermore, RNA-seq analyses at S3 showed that structural genes such as PsC4Hs, PsDFRs, and PsUFGTs in the flavonoid biosynthesis pathway were downregulated in “High Noon,” whereas most PsFLSs, PsF3Hs, and PsF3’Hs were upregulated. Five transcription factor (TF) genes related to flavonoid biosynthesis were also upregulated in “High Noon.” One of these TFs, PsMYB111, was overexpressed in tobacco, which led to increased flavonols but decreased anthocyanins. Dual-luciferase assays further confirmed that PsMYB111 upregulated PsFLS. These results improve our understanding of yellow pigmentation in tree peony and provide a guide for future molecular-assisted breeding experiments in tree peony with novel flower colors.

Keloid-like dermatofibroma

Dermatofibroma is a common benign skin tumor, mainly occurring in young to middle-aged females. It is frequently localized in the lower extremities. A typical dermatofibroma usually presents itself as a single firm papule or nodule, of variable color, bluish, brownish, or pinkish. Its clinical, dermoscopic, and histological features usually allow easy diagnosis [1]. However, it is possible to observe some variations of these typical features. Keloid-like dermatofibroma is one of these atypical presentations rarely reported in the literature [2]. A 40-year-old patient with no previous medical history presented to our dermatology department with a lumbosacral lesion evolving for several months. A physical examination revealed a firm, well-demarcated, asymptomatic erythematous nodule, 5 × 12 mm in size, localized in the lumbosacral area (Fig. 1). The patient denied any trauma preceding the onset of the lesion. There was no personal or familial history of keloidal scars. A dermoscopic examination revealed erythema, telangiectatic vessels, a shiny white streak, and a brownish-yellow pigmentation (Fig. 2). A biopsy was performed. A histological examination revealed an atrophic epidermis. The dermis contained a fibroblastic proliferation of low cell density haphazardly arranged, located on a fibromatous background (Figs. 3 and 4). Dermatofibroma with a keloidal presentation was the diagnosis.

In vitro plant regeneration of Christmas cactus (Schlumbergera truncata (Haw.) Moran) by indirect morphogenesis

Plants of Schlumbergera truncata (Haw.) Moran were obtained by indirect morphogenesis from the segment section of shoots in vitro, they were multiplied and rooted. Also were determined the effect of the lighting regime, the composition of the nutrient medium on the consistency and frequency of callus formation. The studies were conducted during 2016–2018. The mode of effective sterilization (more than 90%) of S. truncata plant explants using 0.1% HgCl2 for 7–8 min was established. Optimal conditions for the induction of callus formation in stem node segments of S. truncata plants (rate more than 90% and significant growth) were created on MS (Murashige and Skoog 1962) nutrient medium supplemented with 1.0 mg/l BAP (6-benzylaminopurine) and 0.3 mg/l NAA (1-naphthylacetic acid) under conditions of placement on the nutrient medium and doing a significant number of cuts on the explants. The light intensity of 2.0–3.0 klx, obtained by a callus of dense consistency of dark green pigmentation, when using the thermostat condition without illumination, the callus had loose consistency, dark yellow pigmentation. It is established that the influence of the lighting regime and the composition of the nutrient medium on the frequency of callus formation is statistically significant. The largest number of shoots was obtained on the MS medium with the addition of 2.0 mg/l of BA. At the same times, shoot proliferation and root induction in such numbers were observed on MS culture medium with the addition of 0.5 mg/l BA and 0.5 mg/l kinetin (multiplication factor – 8.8±0.6 per 60-day cultivation cycle).

Management of Kamala (Jaundice) through Ayurved – A Case Report

Jaundice is a yellow pigmentation of the skin, the conjunctival membrane over the sclera and other mucous membrane caused by hyperbilirubinemia (increase level of bilirubin in blood). Today’s lifestyle with unhygienic and poor dietary habit and alcoholic habits, etc. which are responsible factor to promote hepatic damage which clinically reflect as kamala In this case study 25 years male patient having kamala who was suffering from pain in abdomen, weakness, anorexia, burning maturation and fever on and off, the patient was treated with shodhan chikitsa (Virechan with panchatikta ghrita) followed by shaman chikitsa. Patient got significant result as per the values of bilirubin with symptomatic relief in complaints within 30 days, Kamala can be successfully managed by shodhan and shaman chikitsa , the effect of ayurvedic treatment was assessed in relation to improvement in overall clinical sign and symptoms and biochemical investigations. Further study will be needed as per different assessment criteria.

Isolasi Dan Karakterisasi Bakteri Selulolitik Biopori Sebagai Upaya Awal Percepatan Proses Pengomposan

The purpose of this study was to obtain cellulolytic bacterial isolates isolated from leaf litter in absorption holes biopori FMIPA Unesa and obtain the most optimal cellulolytic bacterial isolates in cellulose degradation. This research was an observational study and the data were analyzed descriptively. Stages of the study began with bacterial isolation, cellulolytic ability testing, and characterization of cellulolytic bacterial isolates. Bacterial isolation was carried out by the pour plate method, isolation was carried out by the streak plate method, cellulolytic testing was carried out using Carboxy Methyl Cellulose media which was given Congo red 1%. While the isolation characterization was done morphologically, physiologically, and biochemically. The results obtained 15 isolates of cellulolytic bacteria that were tested for their ability to degrade cellulose. Cellulolytic test results showed that 6 isolates, namely BS1, BS7, BS10, BS11, BS14, and BS15 had a cellulolytic index of 0.8, 0.8, 0.8, 0.8, 0.8, 0.8. and 1. Isolates BS15 is the most optimal isolate in cellulose degradation with characteristics of the punctiform colony, yellow pigmentation, entire edge, convex elevation, optical opaque and smooth surface, produces catalase enzymes, is non-motile and can ferment in glucose and starch but cannot ferment lactose and also a Gram-negative

PSV-27 Fat color and myoglobin of meat from young cattle fed with mineral or concentrate in growing phase and finished on pasture or feedlot

This trial aimed to evaluate the fat color and myoglobin concentration of three cattle breeds, supplemented in two nutritional plans (NP) during the growing phase and finished in the intensive system on pasture or conventional feedlot. One hundred and nineteen bulls from three genetic groups: Nellore (N), ½ Angus x ½ Nellore (A) and ½ Senepol x ½ Nellore (S), were randomly assigned in two NP on growing phase: mineral (M) or concentrate (C, energy protein 0.3% BW). Following the growing phase, two replicates within each treatment were randomly assigned to one of two finishing systems: pasture (P, n = 59) and conventional feedlot (CF, n = 60). Bulls were supplemented daily with 2% BW of concentrate (16% CP and 78% TDN) and slaughtered at a commercial abattoir. Steaks from longissimus muscle (LM) between 12th and 13th ribs each animal were collected for analysis of concentration myoglobin (MYO) and subcutaneous fat were removed for color analysis (parameters: L*, a*, b*, C* and h*). The data were analyzed by ANOVA using PROC MIXED procedures in SAS 9.4, with a 3×2×2 factorial arrangement. The muscle of Nellore showed greater (P = 0.006) concentration of MYO compared to Angus, however, Senepol had values similar to Nellore and Angus (4.43, 4.03 and 4.25, Nellore, Angus, and Senepol, respectively), and the muscle of animal finished on CF had lower concentration of MYO compared to pasture (P < 0.0001; 4.55 and 3.93, P and CF). The subcutaneous fat of animals finished on pasture showed high (P < 0.05) of L*, a*, b* and C* compared to CF. Therefore, the meat quality parameters were more affected by finished systems, the meat of animals from pasture had the lighter fat and amount of yellow pigmentation, and also presented meat with greater level of myoglobin.

A mutant allele of ζ-carotene isomerase (Z-ISO) is associated with the yellow pigmentation of the “Pinalate” sweet orange mutant and reveals new insights into its role in fruit carotenogenesis

Background Fruit coloration is one of the main quality parameters of Citrus fruit primarily determined by genetic factors. The fruit of ordinary sweet orange (Citrus sinensis) displays a pleasant orange tint due to accumulation of carotenoids, representing β,β-xanthophylls more than 80% of the total content. ‘Pinalate’ is a spontaneous bud mutant, or somatic mutation, derived from sweet orange ‘Navelate’, characterized by yellow fruits due to elevated proportions of upstream carotenes and reduced β,β-xanthophylls, which suggests a biosynthetic blockage at early steps of the carotenoid pathway. Results To identify the molecular basis of ‘Pinalate’ yellow fruit, a complete characterization of carotenoids profile together with transcriptional changes in carotenoid biosynthetic genes were performed in mutant and parental fruits during development and ripening. ‘Pinalate’ fruit showed a distinctive carotenoid profile at all ripening stages, accumulating phytoene, phytofluene and unusual proportions of 9,15,9′-tri-cis- and 9,9′-di-cis-ζ-carotene, while content of downstream carotenoids was significantly decreased. Transcript levels for most of the carotenoid biosynthetic genes showed no alterations in ‘Pinalate’; however, the steady-state level mRNA of ζ-carotene isomerase (Z-ISO), which catalyses the conversion of 9,15,9′-tri-cis- to 9,9′-di-cis-ζ-carotene, was significantly reduced both in ‘Pinalate’ fruit and leaf tissues. Isolation of the ‘Pinalate’ Z-ISO genomic sequence identified a new allele with a single nucleotide insertion at the second exon, which generates an alternative splicing site that alters Z-ISO transcripts encoding non-functional enzyme. Moreover, functional assays of citrus Z-ISO in E.coli showed that light is able to enhance a non-enzymatic isomerization of tri-cis to di-cis-ζ-carotene, which is in agreement with the partial rescue of mutant phenotype when ‘Pinalate’ fruits are highly exposed to light during ripening. Conclusion A single nucleotide insertion has been identified in ‘Pinalate’ Z-ISO gene that results in truncated proteins. This causes a bottleneck in the carotenoid pathway with an unbalanced content of carotenes upstream to β,β-xanthophylls in fruit tissues. In chloroplastic tissues, the effects of Z-ISO alteration are mainly manifested as a reduction in total carotenoid content. Taken together, our results indicate that the spontaneous single nucleotide insertion in Z-ISO is the molecular basis of the yellow pigmentation in ‘Pinalate’ sweet orange and points this isomerase as an essential activity for carotenogenesis in citrus fruits.