Chinensis Plants Research Articles

First report of Tomato Zonate Spot Virus infecting Belamcandachinensis in China.

Tomato zonate spot virus (TZSV, Orthotospovirus tomatozonae, genus Orthotospovirus, family Tospoviridae) was first reported to infect tomato (Solanum lycopersicum) in China in 2008 (Dong et al. 2008). Belamcanda chinensis (L.) Redouté is a perennial herbaceous medicinal plant of the family Iridaceae, which is widely distributed in China. Its rhizome contains abundant active components, mainly including flavonoids, and has antibacterial, anticancer, and antioxidative effects. In July 2023, four B. chinensis plants with virus-like symptoms were collected in Fuyuan County, Yunnan Province in China. The diseased leaves showed chlorosis and ringspots (Fig. S1). Spherical virus particles with a diameter of 80-100 nm were observed in the saps of diseased leaves under a transmission electron microscope (Fig. S2). The presence of an orthotospovirus was confirmed by the previously reported method to amplify the partial sequence (312 nt) of L segment (Huang et al. 2018) (Fig. S3). BLASTn analysis showed that the obtained 312-nt sequence was 95.62% nucleotide identity with TZSV tomato-YN isolate (accession no. NC_010491.1). To obtain the complete genome of this isolate, total RNA from symptomatic leaves of two single diseased B. chinensis were extracted using Hipure Universal RNA Mini Kit (Magen Biotech) and subjected to high-throughput sequencing with a NovaPE150 (Illumina, USA) at MAGIGENE (Shenzhen, China). A total of 41,144,571 clean reads were obtained after removing low quality reads. Quality-controlled, qualified reads were assembled into contigs using Megahit v1.1.2 software. Thirteen contigs shared nucleotide identity ranging 86.94%-97.73% with the L, S, and M segments of TZSV using BLASTn searches online (https://blast.ncbi.nlm.nih.gov/Blast.cgi). In addition, no contigs were mapped to other viral (taxid:10239) and viroidal (taxid:12884) sequences in GenBank Databases. The full-length L, M, and S RNA segments of TZSV-Bc isolate was determined tbe 8917 nt (PP314222), 4718 nt (PP314223) and 3213 nt (PP314224), respectively. These segments were validated by RT-PCR, and Sanger sequencing. They shared nucleotide sequence identities of 95.9%, 97.2%, and 93.1% of the L (NC_010491.1), M (NC_010490.1), and S (NC_010489.1) segments, of the TZSV tomato-YN isolate, respectively. Compared to the TZSV tomato-YN isolate, there exists a missing segment with 113 nt in the intergenic region of S RNA and a segment with 199 nt in M RNA. To further confirm the TZSV infection on B. chinensis, three primers pairs Tosp10/ Tosp11， Tosp5/Tosp6, and NSs-F/NSs-R were tested by RT-PCR for TZSV based on the previous report (Dong et al, 2008). The sequences of amplicons shared >99% nucleotide identity with the corresponding TZSV-Bc isolate sequences. Total of 14 B. chinensis samples were detected with the primer pair N-F/N-R (5'-ATGTCTAACGTCCGGAGTTTAACA-3'/ 5'-AAAAGACAGATCATTGCTGCTCTT-3') by One Step RT-PCR, 6 samples (42.85%) showed the positive results. The mechanical inoculation and RT-PCR detection confirmed TZSV-Bc isolate can infect N. bethamiana. So far, tomato zonate spot virus has been detected in different plants including tobacco (N. tabacum) (Huang et al. 2017), sticktight (Bidens pilosa) (Xu et al. 2022), pepper (Capsicum annuum) (Li et al. 2023) in China. To our knowledge, it is the first report of TZSV naturally infecting B. chinensis plants, which enriches information on the host range of TZSV and will be helpful for disease management.

Enhancing Plant Stress Tolerance: The Role of LcWRKY40 Gene in Drought and Alkaline Salt Resistance in Tobacco and Yeast.

Leymus chinensis, a halophytic perennial grass belonging to the Poaceae family, thrives in saline-alkali grasslands and harbors a rich repository of resistance-related genetic resources. This study focused on deciphering the stress-responsive mechanisms of L. chinensis by conducting transcriptomic sequencing under NaHCO3 stress, which resulted in the annotation of a segment corresponding to the 51WRKY gene. The alkali-induced gene LcWRKY40 (QIG37591) was identified by phylogenetic analysis. Real-time quantitative PCR analysis was performed on L. chinensis plants subjected to PEG6000 and alkaline salt (NaHCO3) stress, and the results indicated that the LcWRKY40 gene was upregulated in both the leaves and roots. The localization of the LcWRKY40 protein was confirmed by the use of green fluorescent protein (GFP) fusion technology in transformed rice protoplast cells. The GAL4-driven transformation of the LcWRKY40 gene in INVScI yeast cells, which exhibited enhanced tolerance upon overexpression of the LcWRKY40 gene under mannitol and alkaline salt (NaHCO3) stress conditions. Under drought stress using mannitol, the fresh weight of Nicotiana tabacum overexpressing the LcWRKY40 gene was significantly higher than that of wild-type(WT) tobacco. Through drought and salt alkali stress, we found that overexpressed tobacco at different stages always outperformed the wild type in terms of fresh weight, SOD, MDA, and Fv/Fm. This study provides preliminary insights into the involvement of the LcWRKY40 gene in responding to drought and alkaline salt stresses, highlighting its role in enhancing plant resistance to drought and saline-alkaline conditions. These findings lay the foundation for future molecular breeding strategies aimed at improving grass resistance from different aspects.

Effect of Nitrogen and Humic Acid on Growth and Yield of Pakcoy (Brassica rapa subsp. chinensis) Plants

Decreased soil quality due to reduced nutrient content is one of the causes of decreasedquality and yield in plants. The use of humic acid organic matter with N fertilizer is one of the effortsto increase nutrients in the soil while increasing plant growth and yield. This research was conductedin Heuleut Village, Leuwimunding District, Majalengka Regency, from May to June 2024. Theresearch method used was the experimental method with a group-randomized design. The treatmentconsisted of 16 treatments that were repeated twice. The variables observed were plant height,number of leaves, root volume, plant fresh weight, leaf area, and plant growth rate. The resultsshowed that the use of N fertilizer at a dose of 115 kg/ha (urea 250 kg/ha) and humic acid 3 g/planthad a good effect on all variables observed. The results of soil analysis showed an increase in N-totaland C-organic.

First Report of Phytopythium litorale Causing Root Rot on Kousa Dogwood in Tennessee and the United States.

Kousa dogwood (Cornus kousa) is an economically important woody ornamental crop that exhibits creamy, white, pointed bracts in late spring, and reddish to pink drupe fruits in late summer and fall. It bears shiny dark green leaves that become reddish-purple to scarlet in the fall. In August of 2023, 3-year-old container grown C. kousa var. chinensis plants in a commercial nursery in Warren Co., Tennessee, exhibited severe yellowing, dieback and root rot symptoms (Fig. 1a and 1b). Dark brown to black lesions were observed in the root and crown region of the plants. Disease severity was 40% to 60% of root area affected, and disease incidence was approximately 40% of 1,000 plants. Surface-sterilized (10% NaOCl: 1 min) symptomatic root tissues were plated on V8-PARPH and incubated at 25°C. Sparse aerial mycelium, showing a distinct rosette or faint radiate to chrysanthemum colony pattern, was observed within four days of incubation (Fig. 2). All isolates produced ovoid or subglose, papillate, and proliferating sporangia in grass blade water cultures (Derviş et al. 2020). Sporangia measured as 19.18 to 24.80 µm X 18.08 to 22.16 µm (n = 50) with a length/width ratio of 1.06 to 1.11. Zoospores observed were between 7.07 to 9.98 µm in diameter (n = 50). Oogonia and oospores were not produced. The ribosomal internal transcribed spacer (ITS) and large subunit (LSU), as well as mitochondrial cytochrome oxidase subunit II (COX-II) genetic markers were amplified and sequenced using primer pairs ITS1/ITS4 (White et al. 1990), NL1/NL4 (Baten et al. 2014), and cox2-F/cox2-RC4 (Choi et al. 2015), respectively. The ITS, LSU, and COX-II sequences of isolates FBG6343, FBG6344 (ITS: PP458373 and PP461387; LSU: PP461390 and PP461391; COXII: PP477112 and PP477113) were 100% identical to those of MN306118, HQ643386, and MN206732, respectively. Based on the morphology (Nechwatal and Mendgen 2006) and sequence data, the isolates were identified as Phytopythium litorale (Nechw.) Abad, De Cock, Bala, Robideau, Lodhi & Lévesque. The pathogenicity test was performed on 3-year-old C. kousa var. chinensis plants grown in a 3-gal container to fulfill Koch's postulates. Kousa dogwood plants were drench inoculated (800 ml/plant) with a pathogen slurry (two plates of 7-day-old culture/liter) of isolates FBG6343 and FBG6364 (five plants per isolate). Control plants were drenched with agar slurry without the pathogen. The study was conducted in a greenhouse maintained at 21 to 23°C and 70% relative humidity with a 16-h photoperiod and irrigated twice a day for 2 min using an overhead irrigation system. Forty-five days after inoculation, plants showed dieback symptoms, and dark brown lesions developed in the roots of all inoculated plants. Isolates with morphology and sequences identical to those of FBG6343 and FBG6364 were recovered from root tissues of all inoculated plants. All control plants remained symptom-free, and P. litorale was not isolated from the root tissue. Previously, P. litorale was reported to cause disease on apple, kiwi, planatus, and rhododendron (Derviş et al. 2020; Li et al. 2021; Mert et al. 2020; Polat et al. 2023). To our knowledge, this is the first report of P. litorale causing root rot of kousa dogwood in Tennessee and the United States. Identification of this pathogen as the causal agent is crucial to developing timely management practices.

First Report of Kiwifruit Root Rot Caused by Globisporangium spinosum in China.

Kiwifruit is widely cultivated for its high vitamin C content and nutritional value. In January 2022, root rot symptoms were found in about 30% of Actinidia chinensis cv. Jinyan plants grafted on A. deliciosa rootstocks in an orchard located in Sanming (26.32°N, 117.23°E), Fujian Province of China. The affected plants appeared stunted, with brown and decaying roots, some of which were covered with white hyphae. To isolate the pathogen, the surfaces of typical symptomatic roots were sterilized for 30 s using 75% ethanol, followed by four rinses in sterile water, placing on potato dextrose agar (PDA), and incubating away from light at 25°C for 7 days. 16 Globisporangium-like isolates were obtained through hyphal tip isolation, displaying a milky-white appearance with irregular protuberances on the surface, and yellow-white backs with radial fold lines. The isolates were then cultured on corn meal agar for 5 days at 25°C in dark for morphological characteristics. Under microscope, the hyphae appeared as long strips without septa and 4.1 to 8.2 µm wide (average 6.7 µm), containing irregularly sized spherical droplets. Both terminal and intercalary hyphae swellings were observed; these appeared either spherical or subspherical, with some having projections. Their dimensions were 12.3 to 27.6 µm (average 17.3 µm). The oospores were mostly spherical, either plerotic or aplerotic, 11.8 to 22.3 µm wide (average 18.9 µm), with occasional projections. The antheridia were rod-shaped and curved, with one end attached to the oogonia. Amplification of the sequences of internal transcribed spacer (ITS) regions and cytochrome c oxidase subunit I (COI) were conducted using the primers ITS1/ITS4 (White et al. 1990) and OomCoxI-Levlo/OomCoxI-Levup (Robideau et al. 2011), respectively. The sequencing results revealed identical ITS and COI sequences in all 16 isolates. BLASTn analysis of the 969-bp ITS sequence ON202808 showed 99.38-99.59% similarity (965/971bp, 967/971bp) with the KJ162353 and AY598701 sequences from Globisporangium spinosum isolates, while the 700-bp COI sequence ON075783 showed 100% and 99.41% identity (680/680bp, 676/680bp) with the GenBank sequences HQ708835 and HQ708832, respectively, from G. spinosum. Phylogenetic analysis also showed that the obtained isolate (termed MA16) clustered with isolates from G. spinosum on the same evolutionary branch. For pathogenicity testing, four-month-old healthy Jinyan (A. chinensis) plants grown in sterilized media were transferred to sterile petri dishes covered with wet filter paper, and their roots were inoculated with a 5-mm-wide disk of MA16 when cultivated on PDA medium for 5 days. Miliang-1 (A. deliciosa) and Hongyang (A. chinensis) plants were treated similarly. The control groups each included three plants that were inoculated with non-colonized PDA. The plants were kept at 25 °C with a 12-/12-h light/dark cycle for 10 days when the inoculated plants exhibited root rot symptoms similar to those seen in the field, together with rotting and browning of the leaves. The control plants appeared healthy with no symptoms. After re-isolated from infected tissues, the pathogen was verified to be G. spinosum according to its ITS sequence, thus fulfilling the Koch's postulates. Recently, Pythium spinosum has been classified as G. spinosum according to whole-genome sequencing and phylogenomic analysis (Nguyen et al. 2022). Based on the morphological features and pathogenicity results, MA16 was identified as G. spinosum (van der Plaats-Niterink 1981; Huo et al. 2023). This report appears to be the first description of kiwifruit root rots caused by G. spinosum in China, and its identification will assist the development of strategies to counteract the disease.

Compatibility of Actinidia arguta with Schisandra chinensis and Malus domestica plants in mixed plantings

Compatibility of Actinidia arguta with Schisandra chinensis and Malus domestica plants in mixed plantings

Decoding early stress signaling waves in living plants using nanosensor multiplexing.

Increased exposure to environmental stresses due to climate change have adversely affected plant growth and productivity. Upon stress, plants activate a signaling cascade, involving multiple molecules like H2O2, and plant hormones such as salicylic acid (SA) leading to resistance or stress adaptation. However, the temporal ordering and composition of the resulting cascade remains largely unknown. In this study we developed a nanosensor for SA and multiplexed it with H2O2 nanosensor for simultaneous monitoring of stress-induced H2O2 and SA signals when Brassica rapa subsp. Chinensis (Pak choi) plants were subjected to distinct stress treatments, namely light, heat, pathogen stress and mechanical wounding. Nanosensors reported distinct dynamics and temporal wave characteristics of H2O2 and SA generation for each stress. Based on these temporal insights, we have formulated a biochemical kinetic model that suggests the early H2O2 waveform encodes information specific to each stress type. These results demonstrate that sensor multiplexing can reveal stress signaling mechanisms in plants, aiding in developing climate-resilient crops and pre-symptomatic stress diagnoses.

First report of Phaeosphaeria caricicola causing leaf spot on Belamcanda chinensis in China.

Belamcanda chinensis (L.) Redouté, a member of the Iridaceae family, is globally well-known for its medicinal value as clearing away heat, detoxifying, detumescence and pain (Qin 2000). In 2021, spots were observed on 40% B. chinensis leaves and about 28 disease index in Wanzhou District (30°32'N; 108°22'E) of Chongqing. Initial symptoms appeared as circular yellow white, sunken spots lesions, and then expanded into irregular lesions, the center of the spots was beige, external layer was light brown and surrounded by yellow halo. Symptomatic leaf tissues (5 × 5 mm) were cut from the infected margin, surface sterilized with 75% ethanol for 1 min, washed with 3% sodium hypochlorite for 3 min, rinsed three times with sterile water, placed on potato dextrose agar (PDA) medium incubated at 25°C for 7 days in the dark, forty isolates with similar morphology were obtained. Three isolates (SG9、SG20 and SG33) was selected for subsequent research. Colonies color changed from beige to light brown color after 14 days on PDA medium. Fungal colonies transformed from beige to brown at the edges after 28 days and light brown on top. Ascomata dark brown, ellipsoidal to globose 116.6 to 253.3 × 89.6 to 172.6 μm in diamensions. Asci stipitate, cylindrical with obtuse ends, and 69.1 to 114.7 × 10.2 to 24.1 μm (n = 30) in size, with eight overlapping linearly biseriate ascospores. Ascospores brown, narrowly fusiform, straight or slightly curved with three transversely septate, slightly constricted at septa, and 9.7 to 12.6 × 27.6 to 32.6 μm (n = 30). These characteristics are consistent with Phaeosphaeria sp. reported by Quaedvlieg et al in 2013. DNA was extracted from representative isolates. The internal transcribed spacer (ITS) region, the large subunit rDNA (LSU), the small subunit rDNA (SSU) and the RNA polymerase II second largest subunit (RPB2) genes were amplified for Polymerase chain reaction (PCR) by used ITS1/ITS4, LR5/LROR, NS1/NS4, and RPB2-5f2/RPB2-7cr primers (White et al. 1990; Vilgalys et al. 1990; Qi M W. et al. 2008; De G. J. et al. 1992). The sequences were submitted to NCBI GenBank: SG-G9 (ITS, OR701701; LSU, OR701699; SSU, OR701700; RPB2, OR738464); SG-G20 (ITS, OQ748032; LSU, OQ780728; SSU, OQ780723; RPB2, OQ779979); SG-G33 (ITS, OQ748033; LSU, OQ780729; SSU, OQ780722; RPB2, OQ779980). A phylogenetic analysis revealed a 99% similarity to the Phaeosphaeria caricicola CBS 603.86 (ITS, KF251182; LSU, GQ387590; SSU, GQ387529; RPB2, KF252189) sequences. Mycelial agar plugs (5-mm diameter) from a 7-day-old PDA culture of a fungal isolate were placed onto pinpricked leaves of three two-year-old B. chinensis plants. While the sterile PDA plugs inoculated in pinpricked leaves of B. chinensis as controls. Inoculated plants were placed in a greenhouse at 25°C and remained 95±1% relative humidity. The inoculated leaves of treatment developed symptoms after 20 days, whereas no symptoms occurred on controls, fulfilling Koch's postulates. The experiments were repeated three times. The fungus was re-isolated and was identical to original isolate by morphologically and molecularly. As far as we know, P. caricicola can cause diseases on carex plants and has been found in Switzerland. This is the first report of P. caricicola causing leaf spot on B. chinensis in China. Along with recording the occurrence of this disease, plant disease management strategies need to be established to reduce losses.

Bacillus velezensis LT1: a potential biocontrol agent for southern blight on Coptis chinensis.

Southern blight, caused by Sclerotium rolfsii, poses a serious threat to the cultivation of Coptis chinensis, a plant with significant medicinal value. The overreliance on fungicides for controlling this pathogen has led to environmental concerns and resistance issues. There is an urgent need for alternative, sustainable disease management strategies. In this study, Bacillus velezensis LT1 was isolated from the rhizosphere soil of diseased C. chinensis plants. Its biocontrol efficacy against S. rolfsii LC1 was evaluated through a confrontation assay. The antimicrobial lipopeptides in the fermentation liquid of B. velezensis LT1 were identified using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS). The effects of B. velezensis LT1 on the mycelial morphology of S. rolfsii LC1 were examined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The confrontation assay indicated that B. velezensis LT1 significantly inhibited the growth of S. rolfsii LC1, with an inhibition efficiency of 78.41%. MALDI-TOF-MS analysis detected the presence of bacillomycin, surfactin, iturin, and fengycin in the fermentation liquid, all known for their antifungal properties. SEM and TEM observations revealed that the mycelial and cellular structures of S. rolfsii LC1 were markedly distorted when exposed to B. velezensis LT1. The findings demonstrate that B. velezensis LT1 has considerable potential as a biocontrol agent against S. rolfsii LC1. The identified lipopeptides likely contribute to the antifungal activity, and the morphological damage to S. rolfsii LC1 suggests a mechanism of action. This study underscores the importance of exploring microbial biocontrol agents as a sustainable alternative to chemical fungicides in the management of plant diseases. Further research into the genetic and functional aspects of B. velezensis LT1 could provide deeper insights into its biocontrol mechanisms and facilitate its application in agriculture.

Nitrate nitrogen enhances the efficiency of photoprotection in Leymus chinensis under drought stress.

Global climate change exerts a significant impact on the nitrogen supply and photosynthesis ability in land-based plants. The photosynthetic capacity of dominant grassland species is important if we are to understand carbon cycling under climate change. Drought stress is one of the major factors limiting plant photosynthesis, and nitrogen (N) is an essential nutrient involved in the photosynthetic activity of leaves. The regulatory mechanisms responsible for the effects of ammonium (NH4 +) and nitrate (NO3 -) on the drought-induced photoinhibition of photosystem II (PSII) in plants have yet to be fully elucidated. Therefore, there is a significant need to gain a better understanding of the role of electron transport in the photoinhibition of PSII. In the present study, we conducted experiments with normal watering (LD), severe drought (MD), and extreme drought (HD) treatments, along with no nitrogen (N0), ammonium (NH4), nitrate (NO3), and mixed nitrogen (NH4NO3) treatments. We analyzed pigment accumulation, reactive oxygen species (ROS) accumulation, photosynthetic enzyme activity, photosystem activity, electron transport, and O-J-I-P kinetics. Analysis showed that increased nitrate application significantly increased the leaf chlorophyll content per unit area (Chlarea) and nitrogen content per unit area (Narea) (p< 0.05). Under HD treatment, ROS levels were lower in NO3-treated plants than in N0 plants, and there was no significant difference in photosynthetic enzyme activity between plants treated with NO3 and NH4NO3. Under drought stress, the maximum photochemical efficiency of PSII (Fv/Fm), PSII electron transport rate (ETR), and effective quantum yield of PSII (φPSII) were significant higher in NO3-treated plants (p< 0.05). Importantly, the K-band and G-band were higher in NO3-treated plants. These results suggest that drought stress hindered the formation of NADPH and ATP in N0 and NH4-treated L. chinensis plants, thus damaging the donor side of the PSII oxygen-evolving complex (OEC). After applying nitrate, higher photosynthetic enzyme and antioxidant enzyme activity not only protected PSII from photodamage under drought stress but also reduced the rate of damage in PSII during the growth of L. chinensis growth under drought stress.

Leaf Spot Caused by Colletotrichum siamense, C. fructicola, and C. aeschynomenes on Ixora chinensis in Guangxi, China.

Ixora chinensis Lam., an important ornamental flower, has become more and more popular in the southwest and southeast regions of China for its bright and abundant flowers (Li et al. 2019). In March 2022, 100% I. chinensis plants showed typical anthracnose symptoms on leaf in Nanning, Guangxi, China (108°22' N, 22°48' E). The central areas of lesions were grayish white with small black particles arranged in a wheel pattern, and the edges of lesions were light red to brown. Three diseased leaf samples were collected from three gardens, respectively. 5×5 mm tissues were cut from infected margins, surface-disinfected in 75% ethanol for 10 s, 2% NaClO for 2 min, rinsed three times in sterilized distilled water, and incubated on PDA at 25°C under 12/12 h light/darkness. Eighty-three morphologically similar colonies were observed on PDA after 5 days, and eight isolates G1-3, G2-1, G3-3, W-1, W-2, LCH2-1, LCH3-3, and LCH4-1 were selected for further study. Genomic DNA of these isolates were extracted from 7-day-old mycelia. Primer pairs ITS1/ITS4, GDF1/GDR1, T1/βt2b, CHS Ⅰ-79F/CHS Ⅰ-354R, CL1/CL2, ACT-512F/ACT-783R, and MAT1-2-1/apn2 were used to amplify ITS loci and GAPDH, CHS-Ⅰ, CAL, ACT, ApMAT genes, respectively (Yang et al. 2009; Silva et al. 2012; Liu et al. 2015). Sequences have been deposited in GenBank (ITS: OQ771884 to OQ771891, GAPDH: OQ759576 to OQ759583, TUB2: OQ759584 to OQ759591, CHS-1: OQ759568 to OQ759575, CAL: OQ759560 to OQ759567, ACT: OQ759552 to OQ759559, ApMat: OQ759544 to OQ759551). Phylogenetic analysis was performed with raxmlGUI v.2.0.0. based on combined sequences of ITS, GAPDH, TUB2, CHS-1, CAL, ACT, and ApMAT using maximum parsimony analysis. The results revealed that isolates G2-1 and W-2 were clustered with Collectrichum fructicola, G3-3, W-1, G1-3, LCH2-1, and LCH3-3 were clustered with C. siamense, and LCH4-1 was clustered with C. aeschynomenes. Three representative isolates W-2, G3-3, and LCH4-1 were selected for morphology and pathogenicity observation. On PDA, the colonies of three isolates presented white to grey cottony mycelia，from the margin to the center, W-2 was white, grey, and light gray, G3-3 showed light gray, white, and grey, LCH4-1 was white and light gray, respectively. Conidia were all hyaline, one-celled, cylindrical, and straight. The conidial sizes of W-2, G3-3, and LCH4-1 were 11.03 to 17.53 × 4.93 to 8.42 μm (n=100), 10.63 to 19.06 × 3.73 to 6.92 μm (n=100), and 11.61 to 20.39 × 3.65 to 6.67 μm (n=100), respectively. Pathogenicity tests of three isolates were conducted on leaves of 1-year-old I. chinensis plants with and without wounds, three plants for each treatment, and five leaves inoculated for each plant. Conidial suspensions (10 µL, 106 conidia/mL in 0.1% sterile Tween 20) were inoculated on each site. Control group was treated with 0.1% sterile Tween 20. All inoculated sites were covered with wet cotton, and all plants were bagged and placed in the greenhouse to maintain humidity at 25℃. After 10 days, all wounded and inoculated leaves showed leaf spot, whereas unwounded and control leaves remained asymptomatic. Koch's postulates were fulfilled by re-isolating the causal agents from diseased leaves. C. siamense and C. aeschynomenes could cause anthracnose of I. chinensis in China (Liu et al. 2016, Li et al. 2021). However, to our knowledge, this is the first report of C. fructicola infecting I. chinensis in China. This study may provide reference for further epidemiological study and prevention of anthracnose on I. chinensis.

The Effect of Giving Tofu Dregs and Manure on The Growth of Pakcoy (Brassica rapa subsp. chinensis) Plants

Pakcoy is a variety of mustard greens whose leaves are used as a vegetable. Pakcoy mustard greens are also often called spoon mustard greens because their shape resembles a spoon. The aim of this research was to determine which factors and chemical combinations had the greatest influence on the growth of pak choy (Brassica rapa subsp. chinensis) plants. Experimental methodology was used in this research's quantitative approach. There were 24 samples used in this research experiment, which included 6 treatments and 4 replications. Statistical analysis of variance (ANOVA) test was used in data analysis for this study. The hypothesis testing process then continues using data from the prerequisite tests, namely ordinariness test and homogeneity test. The results showed that tofu dregs and manure had no effect on the total fresh weight of harvested plants, but did affect plant height and number of leaves at 2 WAP and 6 WAP observations.including the addition of soil and manure, has the best composition for growth in plant height and wet weight of the entire crop.The combination of soil, tofu dregs and manure is a good treatment for increasing the number of leaves.The plant height parameter observed at 2 WAP and 4 WAP in treatments P3 and P1 was the as it were parameter that was altogether diverse between each treatment, based on the findings of the BNT test.

First report of Colletotrichum fioriniae Causing Anthracnose Disease of Rhus chinensis in China.

Rhus chinensis, a tree of major economic importance in China, belongs to the Anacardiaceae. It is the summer host of the aphidMelaphis chinensis which produces a leaf gall utilized for medicinal purposes (Li et al. 2022). In August 2021 and June 2022, dark brown spots were observed on young branches of R. chinensis in Wufeng, Hubei province, China. The plantations of R. chinensis in Wufeng County had different degrees of disease. We focused our survey on three plantations, each with an area of 1.5 hectares and 1600 R. chinensis plants per hectare, and the incidence of the disease was found to be around 70%. Symptoms began as small brown spots that expanded with time and eventually led to large, irregular, dark brown and sunken lesions. Under high temperature and humidity, orange conidiomata appeared on top of the lesions. As the disease progressed, branches rotted, broke, and leaves died and dropped, eventually causing the death of trees. The fungus was isolated from infected branches. Branch pieces were cut and surface disinfested in 75% (v/v) alcohol for 30 sec, then sterilized in 4% sodium hypochlorite for 1 min, and washed three times with sterile distilled water before incubated on potato dextrose agar (PDA) at 25℃.Ten isolates were obtained by a single-spore culturing method, considering HTK-3 isolate showed more pathogenic and grew faster than other isolates, it was selected for further research. After culturing for 7 days on PDA medium, the colony of isolate HTK-3 was cottony with white-to-gray aerial mycelium. The mycelial growth rate was 8.7 mm/day at 25℃. Conidia were single-celled, colorless, smooth-walled, fusiform with acute ends, and measured 7.7 to 14.3 × 3.2 to 5.3μm (mean 11.8 ± 1.3 to 4.2 ± 0.5μm, n = 50). Appressoria were single, medium brown, ovate to ellipsoid, 5.8 to 8.5× 3.7 to 6.1μm (mean 7.2 ± 0.7 × 4.9 ± 0.4 μm, n=50). Microscopic examinations showed conidia of the HTK-3 were hyaline, aseptate, and sub-cylindrical, with obtuse apices and tapering bases. Mycelium of which was hyaline, branched and septate. Based on these morphological features, the fungus was tentatively identified as belonging to Colletotrichum acutatum species complex (Damm et al. 2012). For molecular identification, the ITS region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-1), beta-tubulin 2 (TUB2), and actin (ACT) were amplified and sequenced (Liu et al., 2022). The obtained sequences were deposited into the GenBank [accession numbers: OP630818 (ITS); OP649736 (GAPDH); OP649735 (TUB2); OP649738 (CHS-1); OP649737 (ACT)]. For all of these genes, isolates HTK-3 had a 99-100% similarity to multiple C. fioriniae accessions. A maximum likelihood tree was generated from a multiple sequence alignment of reported isolates (Liu et al. 2022) and HTK-3 was identified as C. fioriniae. To fulfill Koch's postulates, ten healthy branches were inoculated with 5-mm-diameter mycelial plugs of each of ten fungal isolates (Wang et al., 2022). PDAs plug without mycelium was used as control. Six days post-inoculation, all branches developed anthracnose symptoms similar to those observed in the field, while the control remained healthy. The pathogenicity tests were repeated twice with the same results. C. fioriniae was re-isolated from the disease branches and the morphology was consistent with original, fulfilling Koch's postulates. The species of C. fioriniae has been reported to cause severe anthracnose of many species of plants (Eaton et al. 2021). To our knowledge, this is the first report of C. fioriniae as a pathogen of R. chinensis in China. The results will help to target the screening of control agents and provide guidance for disease prevention and control.

How does UV-B radiation influence the photosynthesis and secondary metabolism of Schisandra chinensis leaves?

Schisandra chinensis (Trucz.) is a medicinal plant with a rich profile of physiologically active substances, including lignans. The levels of these substances can be affected by external factors such as light exposure, temperature, and humidity. The current study aimed to explore the impacts of UV-B radiation on the photosynthetic properties and biochemical parameters as well as the contents of major lignans and phenolic compounds in two-year-old S. chinensis plants. The results revealed that exposure to UV-B radiation (10.0 μW cm−2, 30 min per day) did not cause any significant changes in the photosynthetic properties of S. chinensis plants until the 7th or 15th days of treatment. However, a significant reduction in photosynthetic capacity was observed after 30 days of UV-B exposure. The contents of photosynthetic pigments including chlorophyll a, b, total chlorophyll, and carotenoid decreased under UV-B treatment. Besides, we found that the highest level of lignan synthesis, a major bioactive compound in S. chinensis, occurred after seven days of UV-B treatment. However, lignan levels gradually decreased over time, with most individual lignans either having similar or lower contents than the control at 30 days. Furthermore, a comprehensive analysis of 35 phenolic compounds present in S. chinensis leaves showed that quercetin, isoquercetin, and 4-hydroxycinnamic acid were critical phenolic compounds responsible for moderating the plant's response to ultraviolet stress. Consistent with the trend of lignan content, these phenolic compounds exhibited the highest levels after seven days of UV-B treatment and gradually decreased over time. Our findings demonstrate that an appropriate dose of UV-B exposure can stimulate the synthesis of bioactive compounds such as lignans and phenolics via activation of the phenylpropanoid pathway in S. chinensis, enhancing its medicinal value.

Neuroprotective Properties of Berberine: Molecular Mechanisms and Clinical Implications.

Berberine (BBR), an isoquinoline alkaloid natural product, is isolated primarily from Coptis chinensis and other Berberis plants. BBR possesses various bioactivities, including antioxidant, anti-inflammation, anticancer, immune-regulation, and antimicrobial activities. Growing scientific evidence underscores BBR's substantial neuroprotective potential, prompting increased interest and scrutiny. In this comprehensive review, we elucidate the neuroprotective attributes of BBR, delineate the underlying molecular mechanisms, and assess its clinical safety and efficacy. The multifaceted molecular mechanisms responsible for BBR's neuroprotection encompass the attenuation of oxidative stress, mitigation of inflammatory responses, inhibition of apoptotic pathways, facilitation of autophagic processes, and modulation of CYP450 enzyme activities, neurotransmitter levels, and gut microbiota composition. Furthermore, BBR engages numerous signaling pathways, including the PI3K/Akt, NF-κB, AMPK, CREB, Nrf2, and MAPK pathways, to confer its neuroprotective effects. This comprehensive review aims to provide a substantial knowledge base, stimulate broader scientific discourse, and facilitate advancements in the application of BBR for neuroprotection.

Transcriptome profiling reveals multiple regulatory pathways of Tamarix chinensis in response to salt stress.

Multiple regulatory pathways of T. chinensis to salt stress were identified through transcriptome data analysis. Tamarix chinensis (Tamarix chinensis Lour.) is a typical halophyte capable of completing its life cycle in soils with medium to high salinity. However, the mechanisms underlying its resistance to high salt stress are still largely unclear. In this study, transcriptome profiling analyses in different organs of T. chinensis plants in response to salt stress were carried out. A total number of 2280, 689, and 489 differentially expressed genes (DEGs) were, respectively, identified in roots, stems, and leaves, with more DEGs detected in roots than in stems and leaves. Gene Ontology (GO) term analysis revealed that they were significantly enriched in "biological processes" and "molecular functions". Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that "Beta-alanine metabolism" was the most differentially enriched pathway in roots, stems, and leaves. In pair-to-pair comparison of the most differentially enriched pathways, a total of 14 pathways, including 5 pathways in roots and leaves, 6 pathways in roots and stems, and 3 pathways in leaves and stems, were identified. Furthermore, genes encoding transcription factor, such as bHLH, bZIP, HD-Zip, MYB, NAC, WRKY, and genes associated with oxidative stress, starch and sucrose metabolism, and ion homeostasis, were differentially expressed with distinct organ specificity in roots, stems, and leaves. Our findings in this research provide a novel approach for exploring the salt tolerance mechanism of halophytes and identifying new gene targets for the genetic breeding of new plant cultivars with improved resistance to salt stress.

Enrichment of soil nutrients and salt ions with different salinities under Tamarix chinensis shrubs in the Yellow River Delta

In this study, we investigated the characteristics of soil salt ions and soil nutrients at different locations around Tamarix chinensis and with different salinity levels on a coastal beach of the Yellow River Delta. The findings help clarify the effects of T. chinensis growth on the enrichment rates of soil nutrients and salt ions. T. chinensis shrublands with low salinity (0.25 %), moderate salinity (0.51 %), and high salinity (1.03 %) on the coastal beach of the Yellow River Delta were studied. The available nitrogen (AN), available phosphorus (AP), available potassium (AK), total organic carbon (SOC), and levels of eight major salt ions were measured and analyzed at different locations around the T. chinensis plants. The results showed that soil salinity significantly affected the contents of soil nutrients and salt ions. Soil salinity, soil location, and their interaction had significant effects on the following contents of soil: AN, AP, SOC, K+, Na+, Cl−, and SO42− (P < 0.01). The soil nutrient contents differed significantly at different locations around a T. chinensis plant (P < 0.05), and the highest nutrient content was found under the center of the canopy, followed by that at the roots. The soil nutrient enrichment rates under different salinity levels were all greater than 1, and T. chinensis shrubs had the strongest enrichment capacity for SOC. The enrichment rates of AN, AK, and SOC under the center and edge of the canopy of T. chinensis were higher under moderate salinity than under low or high salinity; thus, T. chinensis produced a stronger “fertile island” effect under moderate salinity. Soil salinity and location had a significant impact on the enrichment of salt ions. Under low and moderate salinity, the enrichment rates of soil K+, Ca2+, and Mg2+ were all greater than 1, forming “salt islands,” whereas under high salinity, “salt basin” formed. In soil at different locations and with different salinity levels, T. chinensis produced a “salt basin” effect on Na+ and Cl− but a “salt island” effect on soil SO42− and HCO3–. The Na+ enrichment rates under low and moderate salinity were significantly lower than those under high salinity. In summary, T. chinensis shrubs under different salinity levels in the coastal beaches all exhibited the fertile island effect, which was most significant under moderate salinity. In the saline–alkali land on coastal beaches mainly containing Na+ and Cl−, T. chinensis had a salt basin effect on both ions; thus, T. chinensis shrubs significantly reduced the overall soil salinity.

First report of Alternaria alternata causing leaf blight on Trollius chinensis in China.

Trollius chinensis is widely distributed in east Asian countries that include China, Siberia, and Japan, with antibacterial, antiviral, anti-inflammatory and analgesic activity for medical applications. In August 2021, leaf blight was observed on nearly 80~95% of T. chinensis plants growing in Daxinganling (51.43°N, 126.39°E) from Heilongjiang Province, China. Initial symptoms were gray-black necrosis, wilting progressing from the leaf margin, and eventual defoliation. Six T. chinensis plants with typical symptoms were randomly collected, and three fresh leaf samples were collected from each plant. Diseased leaf pieces that measured 5 mm square were disinfected in 75% ethyl alcohol for 30 s and 7% NaClO for 60 s, rinsed three times in sterile distilled water, and placed on potato dextrose agar (PDA). Twelve fungal isolates, obtained by single-spore isolations, were selected for further. These isolates produced colonies that measured 63 to 73 mm in diameter after 7 days growth on PDA. Colonies were black to brown in color with gray-white aerial hyphae on their surfaces, neat edges, olive green on the back. The isolates produced conidia that were ovate to pear-shaped, brown to black in color, with 1 to 4 transverse septa and 0 to 1 oblique septa, smooth surfaced, parietal cells extending into the beak, and measured 12.5 to 37.5 × 5.0 to 12.5 μm(n=150). Conidiophores were dark, erect or curved, branched, with pronounced spore marks, and measured 35.0 to 50.0 × 4.0 to 5.0 μm(n=150). All twelve fungal isolates were morphologically similar to Alternaria alternata (Simmons 2007). Two representative isolates jlh01 and jlh02 were used for molecular identification. The internal transcribed spacer (ITS) region, RNA polymerase second largest subunit (RPB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), translation elongation factor 1-alpha (TEF1), and Alternaria major allergen (Alt a 1) were amplified with the primers ITS4/ITS5, RPB2-5F2/RPB2-7CR (Khodaei and Arzanlou 2013), gpd1/gpd2, EF1-728F/EF1-986R (Nishikawa and Nakashima 2020) and Alt-for/Alt-rev (Woudenberg et al.2015). The resulting sequences were deposited in GenBank (ITS, OM095427, OM108099; RPB2, OM131213, OM131214; GAPDH, OM201165, OM201166; TEF1, OM131211, OM131212; Alta1, OM201167, OM201168). Phylogenetic tree results showed 100% similarity between jlh01, jlh02 and the type strain CBS 118812. Morphological and molecular analysis results confirmed the identity of the fungus as A. alternata. Pathogenicity tests were done by spraying water-spore suspensions containing 106 spores per ml of A. alternata isolates jlh01 and jlh02 on leaves of six healthy T. chinensis plants, separately. Six control plants were sprayed with distilled water and both sets of plants covered with plastic bags and placed in a greenhouse maintained at 25⁰ C. Plastic bags were removed from all plants after 48 h. Black brown lesions and concentric rings developed on spore-inoculated plants after 15 days and control plants remained symptomless. The pathogenicity tests were conducted three times. A. alternata was reisolated and identified based on morphological and molecular traits, thus fulfilling Koch's postulates. To our knowledge, this is the first report of A. alternata causing leaf blight on T. chinensis in China. Based on the plant's medicinal value, this report provides the basis for further research and control of T. chinensis leaf blight.

First report of Ilyonectria robusta causing root rot on Coptis chinensis in Chongqing, China.

Coptis chinensis belongs to the Ranunculaceae family and is a widely used traditional Chinese herb. Chongqing Municipality produces >60% of China's production. Root rot seriously reduced yield and quality (Mei et al. 2021). In May 2020, root rot of C. chinensis were observed on 3-year-old roots with an average incidence of 45.3% in three commercial fields (about 0.5 acre) in Fengmu Town, Shizhu County (30.24°N; 108.48°E) from Chongqing. Diseased plants were stunted and less vigorous with wilting and twisting leaves. Brown or black discoloration lesion was appeared in the vascular and cortical tissue of roots and rhizomes. Ten fresh symptomatic plants were randomly sampled from the fields. Root tissues were surface sterilized in 75% ethanol for 60s, rinsed thrice with sterile water, placed on potato dextrose agar (PDA), and incubated at 25°C for 7 days. A total of 11 isolates were obtained from the infected tissues. Pure colonies of all fungal isolates had similar characteristics, and five isolates (a2, a4, a9, a11, a12) were randomly selected for further study. Colonies of this fungus were aurantium and felty at first, and then became brownish grey. Macroconidia (n=50) were predominating, hyaline, cylindrical, predominantly straight with both ends broadly rounded, 1~3 septate; one septate, 18.8~25.5×5.9~6.8μm; two septate, 22.6~35.4×6.1~7.2μm; three septate, 26.1~42.5×7.2~8.0 μm. Microconidia (n=50) were hyaline, ellipsoid to ovoid, 0 to 1 septate; aseptate, 7.5~8.8×3.4~4.3μm. Chlamydospores (n=50) were hyaline at first, and becoming brown, globose to subglobose, smooth, 8.3~12.5×8.1~13.5μm, mostly occurring intercalary in chains. The DNA of isolates were extracted and the ITS, HIS, TEF, TUB2 genes were amplified and sequenced using the primers ITS1/ITS4, CYLH3F/CYLH3R, EF1/EF2, T1/CYLTUB1R, respectively (Cabral et al. 2012). The representative isolate a2 were deposited in GenBenk (OK105140, ITS; OM799544, HIS; OK493444, TEF; OK493445, TUB2). BLAST analysis showed the ITS, HIS, TEF, TUB2 sequences of a2 were 100% (417/417), 100% (472/472), 100% (762/762), and 99.7% (490/491) homology with those of Ilyonectria robusta (CBS 605.92) from Tilia petiolaris in Germany. Phylogenetic analysis using Maximum Likelihood and concatenated sequences (ITS+HIS+TEF+TUB2) with MEGA7 placed isolate a2 in I. robusta with 100% bootstrap support. The isolate was thus identified as I. robusta based on morphological and molecular characteristics (Cabral et al. 2012). Thirty healthy 6-month-old C. chinensis plants were used for the pathogenicity tests, and five plants were into each of 6 pots. 10ml of conidia suspension (1×106conidia/ml) of 10-day-old isolate a2 was gently applied to the soil in each of 6 pots. Sterile water (10ml) was applied to each of 6 pots as control. All 12 pots were placed in a greenhouse (25°C, 12h photoperiod). After 6 weeks inoculation, all inoculated plants showed twisting and wilting symptoms, and the roots showed light-brown to dark-brown lesions. No symptoms were observed on the controls. The pathogen was reisolated from all symptomatic roots and identified as I.robustaas previously described above. The test was repeated twice with similar results. Although this fungus was previously reported to cause root disease on many plants (Zheng et al. 2022; Qiao et al. 2019; Guggenheim et al. 2019), this is the first report of I. robusta causing root rot on C. chinensis in China, and will establish a foundation for controlling the disease.

Effects of a compound Trichoderma agent on Coptis chinensis growth, nutrients, enzyme activity, and microbial community of rhizosphere soil.

Root rot diseases are prevalent in many Coptis chinensis Franch. production areas, perhaps partially due to the overuse of synthetic fertilizers. Synthetic fertilizers can also lead to soil degradation. Trichoderma is widely used in biofertilizers and biopesticides. This study applied a combination of four Trichoderma species (compound Trichoderma agent, CTA) to C. chinensis and evaluated its effects on growth, as well as rhizosphere soil nutrients, enzyme activities, and microbial community structure. The purpose of this study was to estimate the potential of using CTA as a biofertilizer for C. chinensis, and determine if it could, at least partially, replace synthetic fertilizers to control root rot disease and maintain soil fertility. CTA, compound fertilizer and sterile water were applied to C. chinensis plants. After 60 days, the soluble sugar, soluble protein, chlorophyll of leaves, and individual weight of each plant were measured. The rhizosphere soil nutrient content, enzymatic activity, and the microbial community were also determined. The results were analyzed to evaluate the effect of CTA on C. chinensis growth and soil fertility. CTA increased the soluble protein, chlorophyll, and individual weight of C. chinensis plants while compound fertilizer decreased chlorophyll. CTA increased the activities of urease and catalase in rhizosphere soil, whereas the compound fertilizer decreased urease, catalase, and alkaline phosphatase activities. CTA elevated soil pH, while compound fertilizer reduced it. CTA had no significant effects on soil nutrients and organic matter. CTA decreased the fungal number and alpha-diversity of fungi and bacteria, and both the fungal and bacterial communities were significantly different from the other two. CTA increased B/F value, which improved the rhizosphere microbial community. Both CTA and the compound fertilizer significantly altered the soil microbial community. The relative abundance of Ascomycota was higher and Basidiomycota was lower after CTA treatment than after the other two treatments, indicating that the soil treated with CTA was healthier than that of the other two treatments. CTA decreased harmful Ilyonectria mors-panacis and Corynebacterium sp. And increased beneficial Ralstonia picketti. Trichoderma spp. could exist in C. chinensis rhizosphere soil for a long time. The functional prediction results demonstrated that CTA reduced some rhizosphere phytopathogenic fungi. Correlation analysis showed that CTA elevated rhizosphere pH and enzyme activities. In summary, synthetic fertilizers damaged soil fertility, and the overuse of them might be responsible for root rot disease, while CTA could promote C. chinensis growth, improve soil and decrease the incidence and severity of C. chinensis root rot disease. Therefore, as a biofertilizer, CTA can, at least partially, replace synthetic fertilizers in C. chinensis production. Combining it with organic fertilizer will increase the potential of Trichoderma.