First Report of Alternaria Leaf Spot Caused by Alternaria alternata on Ulmus parvifolia in Jiangsu, China.

Ulmus parvifolia Jacq. is an important tree with ornamental value, which is widely planted in Hebei and southern regions of China. In September 2022, a leaf spot symptom was observed on about approximately 20% U. parvifolia seedlings growing a tree farm (20000 m2) of Jiangsu Academy of Forestry (118°45'57.30″E, 31°51'27. 94″N). Gray to black spots appeared on leaves of seedlings. Five diseased leaves were collected from five different seedlings. The pieces were excised from the margins between healthy and diseased tissues, surface sterilized in 75% ethanol for 30 s and then in 1.5% NaClO for 90 s, rinsed three times in sterilized distilled water, plated on potato dextrose agar (PDA) and incubated at 25℃ in the darkness. Pure cultures were obtained by monosporic isolation. Six isolates with identical morphological features and the internal transcribed spacer (ITS) sequences were obtained (the isolate rate of 67%), and identified as Alternaria sp. A representative isolate, LY-1-1 was used for the further study. The colony of LY-1-1, growing on PDA was cotton-like and brown in color with gray-white aerial hyphae on their surfaces, and its reverse was dark grey. The conidia were ovate to pear-shaped, brown in color, with 1 to 4 transverse septa and 0 to 1 longitudinal septa, parietal cells extending into the beak, and measured 7.1 to 12.5×3.8 to 7.1 µm (n=35). These characteristics were consistent with the description of Alternaria sp. (Simmons 2007). The regions of ITS, large subunit ribosomal RNA (LSU), small subunit ribosomal RNA (SSU), anonymous region OPA10-2 genomic sequence (OPA10-2), Alternaria 1 major allergen (Alta1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and translation elongation factor 1-alpha (TEF1) genes (GenBank Accession No. OR047916, OR051904, OR047919, OR061065, OR061063, OR061064, and OR061062, respectively) were amplified (White et al. 1990; Woudenberg et al. 2015) and sequenced. These obtained sequences showed 99.86-100% similarity to the ITS (514/515 bp) of A. alternata isolate SPM-2 (OR378581), LSU (801/801 bp) of isolate B9 (OR366492), SSU (1019/1020 bp) of strain LSU0766 (MT000349), OPA10-2 (632/633 bp) of strain 19-1 (MN185000), Alta1 (470/470 bp) of strain CMML21-73 (OQ831518), GAPDH (177/177 bp) of isolate CS36-3 (KY814638), and TEF1 (240/240 bp) of isolate SY-6 (OP980553). A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7. The isolate LY-1-1 clustered in the A. alternata clade with 98% bootstrap support. Three 3-month-old U. parvifolia seedlings were wounded with a sterile needle and inoculated with 20 µL conidia suspension (1×106 spores/mL) on the left sides of leaves. Inoculation on the right side with 20 µL of sterile water was treated as a control. All inoculated plants were incubated in a greenhouse at 25℃, 80% relative humidity, and a 12-h light/dark cycle. The experiment was repeated three times. After 5 days of inoculation, typical gray to black spots were found on the left sides of all inoculated leaves, and the control did not have any leaf spot symptoms. Subsequently, the same fungus was reisolated and identified based on morphological and molecular traits, fulfilling Koch's postulates. The A. alternata has been reported to cause leaf spot on pecan (Wu et al. 2020), fruit spot on olive (Alam et al. 2019) and fruit rot on lychee (Alam et al. 2017). However, there are no other reports of A. alternata on U. parvifolia in the world. Thus, this study provides an important reference for the biology, epidemiology of A. alternata.

Lipidomic and comparative transcriptomic analysis of fatty acid synthesis pathway in Carya illinoinensis embryo.

Pecan (Carya illinoinensis (Wagenh.) K. Koch) is an important oilseed nut and is rich in fatty acids (FAs) and flavonols. Pecan FA has significantly edible, industrial and clinical value. To investigate the dynamic patterns and compositions of FA, and the molecular mechanism that controls FA accumulation in pecan, lipidomic and transcriptomic analyses were performed to determine lipid profiles and gene expression in pecan's FA biosynthesis pathway. In the present study, compared with cultivars 'Caddo' and 'Y-01', 'Mahan' formed larger and heavier embryos and accumulated higher oil content. Lipidomic analysis showed that FA and (O-acyl)-1-hydroxy FA contents were higher in 'Mahan' at the mature stage. Based on full-length and comparative RNA-Seq, differential expression gene enrichment analysis revealed that many functional genes participated in the pathways of 'fatty acid biosynthesis', 'fatty acid metabolism' and 'linoleic acid metabolism'. High FA accumulation model from 'Mahan' demonstrated that key enzyme-encoding genes played an important role in regulating FA biosynthesis. Co-expression module analysis indicated that several transcription factors (TFs; MYB, TCP, bHLH, Dof, ERF, NAC) were involved in FA accumulation by regulating the expression of functional genes, and real-time quantitative PCR verification proved that these TFs had a high correlation with the pecan FA accumulation pattern. These findings provided an insight into the molecular mechanism of FA accumulation in C. illinoinensis embryo, which contributes to pecan oil yielding and pecan molecular breeding.

Ribosome footprint profiling enables elucidating the systemic regulation of fatty acid accumulation in Acer truncatum.

BACKGROUND: The accumulation of fatty acids in plants covers a wide range of functions in plant physiology and thereby affects adaptations and characteristics of species. As the famous woody oilseed crop, Acer truncatum accumulates unsaturated fatty acids and could serve as the model to understand the regulation and trait formation in oil-accumulation crops. Here, we performed Ribosome footprint profiling combing with a multi-omics strategy towards vital time points during seed development, and finally constructed systematic profiling from transcription to proteomes. Additionally, we characterized the small open reading frames (ORFs) and revealed that the translational efficiencies of focused genes were highly influenced by their sequence features. RESULTS: The comprehensive multi-omics analysis of lipid metabolism was conducted in A. truncatum. We applied the Ribo-seq and RNA-seq techniques, and the analyses of transcriptional and translational profiles of seeds collected at 85 and 115 DAF were compared. Key members of biosynthesis-related structural genes (LACS, FAD2, FAD3, and KCS) were characterized fully. More meaningfully, the regulators (MYB, ABI, bZIP, and Dof) were identified and revealed to affect lipid biosynthesis via post-translational regulations. The translational features results showed that translation efficiency tended to be lower for the genes with a translated uORF than for the genes with a non-translated uORF. They provide new insights into the global mechanisms underlying the developmental regulation of lipid metabolism. CONCLUSIONS: We performed Ribosome footprint profiling combing with a multi-omics strategy in A. truncatum seed development, which provides an example of the use of Ribosome footprint profiling in deciphering the complex regulation network and will be useful for elucidating the metabolism of A. truncatum seed oil and the regulatory mechanisms.

Characterization and phylogenetic relationships analysis of the complete chloroplast genome of Carya ovata.

Carya ovata is a slow-growing, long-lived deciduous species that belongs to section Carya of genus Carya. In this study, we de novo assembled the complete chloroplast genome of C. ovata, and analyzed its phylogenetic relationship. The circular genome was 160,765 bp in length, comprising a large single-copy region (89,975 bp), a small single-copy region (18,788 bp), and a pair of inverted repeat regions (26,001 bp each). The chloroplast genome was predicted to contain 131 genes, including 83 protein-coding genes, 40 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. Overall, the GC content of the chloroplast genome was 36.16%. Phylogenetic analysis suggested that C. ovata was closely related to C. illinoinensis, a representative of section Apocarya within the genus Carya.

The complete chloroplast genome of Ulmus parvifolia, an important landscaping tree.

Ulmus parvifolia is a promising tree species for landscaping. In this study, the complete genome of U. parvifolia was reported using next-generation sequencing technology. The chloroplast genome was a circular double-stranded DNA molecule with 159,182 bp in length. It contained a large single copy (LSC) region of 87,838 bp, a small single copy (SSC) region of 18,750 bp, and two inverted repeat (IRa and IRb) regions of 26,297 bp each, which exhibited a typical quadripartite structure. A total of 133 genes were identified, including 84 protein-coding genes, 41 tRNA genes, and eight rRNA genes. The overall GC content in the chloroplast genome was 35.59%. Phylogenetic analysis indicated that U. parvifolia, as a representative of Sect. Microptelea within the Ulmus genus, is sister to the species of Sect. Ulmus.

Characterization of the complete chloroplast genome sequence of Koelreuteria bipinnata.

Koelreuteria bipinnata is an important ornamental tree with attractive flowers and fruits. In this study, we used next-generation sequencing technology to obtain the complete chloroplast genome of K. bipinnata. The entire genome was determined to be 163,863 bp in size, harboring a typical quadripartite structure with a large single copy (LSC) region of 90,240 bp, a small single copy (SSC) region of 18,883 bp, and a pair of 27,370 bp inverted repeat (IR) regions. The genome was predicted to contain 132 genes, including 84 protein-coding genes, 40 tRNA genes, and 8 rRNA genes. The overall GC content of K. bipinnata chloroplast genome was 37.29%. Phylogenetic analysis based on complete chloroplast genome sequences indicated that K. bipinnata was closely related to K. paniculate. This study would be useful for future population genetics studies and phylogenetic analysis of K. bipinnata.