Autophagy functions as a cytoprotective mechanism by regulating programmed cell death during desiccation in Syntrichia caninervis.

Desiccation is a state of extreme water loss that is lethal to many plant species. Some desert plants have evolved unique strategies to cope with desiccation stress in their natural environment. Here we present the remarkable stress management mechanism of Syntrichia caninervis, a desert moss species which exhibits an 'A' category of desiccation tolerance. Our research demonstrated that desiccation stress triggers autophagy in S. caninervis while inhibiting Programmed Cell Death (PCD). Silencing of two autophagy-related genes, ATG6 and ATG2, in S. caninervis promoted PCD. Desiccation treatment accelerated cell death in ATG6 and ATG2 gene-silenced S. caninervis. Notably, trehalose was not detected during desiccation, and exogenous application of trehalose cannot activate autophagy. These results suggested that S. caninervis is independent of trehalose accumulation to triggered autophagy. Our results showed that autophagy function as prosurvival mechanism to enhance desiccation tolerance of S. caninervis. Our findings enrich the knowledge of the role of autophagy in plant stress response and may provide new insight into understanding of plant desiccation tolerance.

Metabolomic Analysis of the Desert Moss Syntrichia caninervis Provides Insights into Plant Dehydration and Rehydration Response.

Desiccation-tolerant (DT) plants can survive extreme dehydration and tolerate the loss of up to 95% of their water content, making them ideal systems to determine the mechanism behind extreme drought stress and identify potential approaches for developing drought-tolerant crops. The desert moss Syntrichia caninervis is an emerging model for extreme desiccation tolerance that has benefited from high-throughput sequencing analyses, allowing identification of stress-tolerant genes; however, its metabolic response to desiccation is unknown. A liquid chromatography-mass spectrometry analysis of S. caninervis at six dehydration-rehydration stages revealed 912 differentially abundant compounds, belonging to 93 metabolic classes. Many (256) metabolites accumulated during rehydration in S. caninervis, whereas only 71 accumulated during the dehydration period, in contrast to the pattern observed in vascular DT plants. During dehydration, nitrogenous amino acids (l-glutamic acid and cysteinylglycine), alkaloids (vinleurosine) and steroids (physalin D) accumulated, whereas glucose 6-phosphate decreased. During rehydration, γ-aminobutyric acid, glucose 6-phosphate and flavonoids (karanjin and aromadendrin) accumulated, as did the plant hormones 12-oxo phytodienoic acid (12-OPDA) and trans-zeatin riboside. The contents ofl-arginine, maltose, turanose, lactulose and sucrose remained high throughout dehydration-rehydration. Syntrichia caninervis thus accumulates antioxidants to scavenge reactive oxygen species, accumulating nitrogenous amino acids and cytoprotective metabolites and decreasing energy metabolism to enter a protective state from dehydration-induced damage. During subsequent rehydration, many metabolites rapidly accumulated to prevent oxidative stress and restore physiological activities while repairing cells, representing a more elaborate rehydration repair mechanism than vascular DT plants, with a faster and greater accumulation of metabolites. This metabolic kinetics analysis in S. caninervis deepens our understanding of its dehydration mechanisms and provides new insights into the different strategies of plant responses to dehydration and rehydration.

Transcriptional profiling analysis providing insights into desiccation tolerance mechanisms of the desert moss Syntrichia caninervis.

Syntrichia caninervis is a desiccation tolerant moss and is the dominant bryophyte found in biological soil crusts in the Gurbantunggut desert. In this study, we assessed the transcriptome profiles of S. caninervis gametophytes during the dehydration-rehydration (D-R) process (across 9 time points) using Illumina sequencing. In total, 22489 transcripts were identified, including 5337 novel transcripts, that mapped to the reference genome. A total of 12548 transcripts exhibited significant alterations in the D-R samples compared with the control samples. The differentially expressed transcripts (DETs) possessed several enriched Gene Ontology terms, such as "water stress response", "oxidation-reduction process", "membrane metabolism", "photosynthesis", and "transcription factor activity". Moreover, during early dehydration stress, the DETs were significantly enriched in stress-related pathways from the Kyoto Encyclopedia of Genes and Genomes, such as "phenylpropanoid biosynthesis", "alpha-linolenic acid metabolism", and "fructose and mannose metabolism". Photosynthesis-related transcripts (e.g., ScPsa H, ScRubisco, and ScLhcb1) were inhibited during the dehydration treatment and significantly accumulated during the late rehydration period. Most transcripts from the late embryogenesis abundant proteins (LEA) and early light-inducible protein (ELIP) families strongly accumulated at the late dehydration stage. These pathways were positively correlated with the content changes of absolute water content and Fv/Fm values, alongside peroxidase and superoxide dismutase activities. Seven transcription factor families, including AP2-ERF, bHLH, G2-like, MYB, NAC, WRKY, and bZIP, were enriched in DETs during D-R treatment. This study is the first transcriptome analysis using the S. caninervis genome for gene annotation and multigroup D-R treatment points. Our results demonstrated the detailed dynamic changes in the transcriptome of S. caninervis during the D-R process. These results also improve understanding of desiccation tolerant plants' adaptations to desiccation stress at the transcription level and provide promising gene resources for transgenic crop breeding.

Genome-Wide Characterization and Expression Profiling of ABA Biosynthesis Genes in a Desert Moss Syntrichia caninervis.

Syntrichia caninervis can survive under 80-90% protoplasmic water losses, and it is a model plant in desiccation tolerance research. A previous study has revealed that S. caninervis would accumulate ABA under dehydration stress, while the ABA biosynthesis genes in S. caninervis are still unknown. This study identified one ScABA1, two ScABA4s, five ScNCEDs, twenty-nine ScABA2s, one ScABA3, and four ScAAOs genes, indicating that the ABA biosynthesis genes were complete in S. caninervis. Gene location analysis showed that the ABA biosynthesis genes were evenly distributed in chromosomes but were not allocated to sex chromosomes. Collinear analysis revealed that ScABA1, ScNCED, and ScABA2 had homologous genes in Physcomitrella patens. RT-qPCR detection found that all of the ABA biosynthesis genes responded to abiotic stress; it further indicated that ABA plays an important role in S. caninervis. Moreover, the ABA biosynthesis genes in 19 representative plants were compared to study their phylogenetic and conserved motifs; the results suggested that the ABA biosynthesis genes were closely associated with plant taxa, but these genes had the same conserved domain in each plant. In contrast, there is a huge variation in the exon number between different plant taxa; it revealed that ABA biosynthesis gene structures are closely related to plant taxa. Above all, this study provides strong evidence demonstrating that ABA biosynthesis genes were conserved in the plant kingdom and deepens our understanding of the evolution of the phytohormone ABA.

ScATG8 Gene Cloned from Desert Moss Syntrichia caninervis Exhibits Multiple Stress Tolerance.

Syntrichia caninervis is the dominant species of biological soil crust in the desert, including the Gurbantunggut Desert in China. It is widely distributed in drylands and considered to be a new model of vegetative desiccation tolerance moss. Here, we cloned an ATG8 gene from S. caninervis and confirmed its function under multiple abiotic stresses, both in situ and in Physcomitrium patens. The results showed that the ScATG8 gene encoded a protein with a highly conserved ATG8 functional domain. ScATG8 gene was increasingly expressed under different abiotic stresses. Under desiccation stress, the overexpression of ScATG8 enhanced the tolerance of S. caninervis and its ability to scavenge ROS. In addition, ScATG8 overexpression promoted the growth of P. patens under multiple stress conditions. Thus, ScATG8 may be a multifunctional gene, and it plays a critical role in the survival of S. caninervis under various abiotic stresses. Our results provide new insights into the function of ATG8 in enabling desiccation tolerance and open up more possibilities for subsequent plant molecular breeding and the mining of the resistance genes of S. caninervis and other moss species.

BaZFP1, a C2H2 Subfamily Gene in Desiccation-Tolerant Moss Bryum argenteum, Positively Regulates Growth and Development in Arabidopsis and Mosses.

C2H2 zinc finger protein (C2H2-ZFP) plays an important role in regulating plant growth, development, and response to abiotic stress. To date, there have been no analyses of the C2H2-ZFP family in desiccation-tolerant moss. In this study, we identified 57 BaZFP transcripts across the Bryum argenteum (B. argenteum) transcriptome. The BaZFP proteins were phylogenetically divided into four groups (I-IV). Additionally, we studied the BaZFP1 gene, which is a nuclear C2H2-ZFP and acts as a positive regulator of growth and development in both moss and Arabidopsis thaliana. The complete coding sequence of the BaZFP1 gene was isolated from B. argenteum cDNA, which showed a high expression level in a dehydration-rehydration treatment process. The overexpression of the BaZFP1 gene in the Physcomitrium patens and B. argenteum promoted differentiation and growth of gametophytes. Heterologous expression in Arabidopsis regulated the whole growth and development cycle. In addition, we quantitatively analyzed the genes related to growth and development in transgenic moss and Arabidopsis, including HLS1, HY5, ANT, LFY, FT, EIN3, MUS, APB4, SEC6, and STM1, and found that their expression levels changed significantly. This study may pave the way for substantial insights into the role of C2H2-ZFPs in plants as well as suggest appropriate candidate genes for crop breeding.

Overview of heavy metal pollution and health risk assessment of urban soils in Yangtze River Economic Belt, China.

Using national multi-purpose regional geochemical survey (NMPRGS) data, this overview systematically summarizes the pollution levels and health risks posed by 8 heavy metals (As, Cd, Pb, Zn, Hg, Cu, Cr and Ni) in urban soils of 86 cities above the prefecture level in Yangtze River Economic Belt (YREB). Meanwhile, the spatial distribution and main sources of heavy metal pollutants in urban soils of key cities are described in detail. On a regional scale, Hg and Cd contamination in urban soils of YREB is most prominent, and As, Cu, Pb and Zn contamination exists in several cities, while Cr and Ni contamination is almost not shown. The type of industrialization and the history of urbanization affect the soil heavy metal pollution in majority cities, and the influence of geological background on the content of heavy metals in urban soils cannot be ignored. Specifically, the urban pollution of Cd, As, Pb and Zn is mainly concentrated in Hunan and Hubei Provinces, which are highly developed in mining industry, especially in Zhuzhou, Chenzhou, Huangshi and Xiangtan cities, while the major soil Hg pollution occurs in Zhejiang and Jiangsu Provinces with rapid economic development, where Shaoxing, Ningbo, Suzhou and Wuxi are the key polluted cities. Heavy metals in the urban soils investigated generally posed low non-carcinogenic and carcinogenic risks to the adults. However, the non-carcinogenic risk to minors in some cities (e.g., Chenzhou and Huangshi) should be given cautious attention.

[Migration, Transformation Characteristics, and Ecological Risk Evaluation of Heavy Metal Fractions in Cultivated Soil Profiles in a Typical Carbonate-Covered Area].

In China, high heavy metal concentrations in cultivated soil are mainly distributed in carbonate-covered areas. The migration and transformation of heavy metals in such soils are influenced by interactions between natural processes and human activities. This study examined the profiles of nine paddy soils, derived from carbonate rocks in Guangxi. The Cd, As, Zn, Cr, Cu, Hg, Ni, and Pb contents we determined, and soil properties such as pH, Corg content, and fractions of Cd, As, Zn, and Cr were tested. Based on the above data, we assessed the vertical distribution of heavy metal fractions, as well as the ecological risks and factors affecting the migration ability of heavy metals, under the influence of human activities and natural soil formation. The results show that compared with the carbonate rocks in Guangxi, the soil profile of the study area is significantly enriched with all eight heavy metals. Among them, Cd, As, Zn, and Cr exceeded China's agricultural land (paddy field) pollution risk screening values, and Cd and As partially exceed the risk intervention values. All fractions of Cd in the soil profiles are widely distributed. The proportion of water-soluble and ion-exchange Cd fractions with high ecological risk decreased significantly from the surface to greater depths in the soil profile. As, Zn, and Cr in the soil profile were mainly in residual states, and the proportion of water-soluble and ion-exchange fractions did not change considerably with increasing depth. The evaluation results of RAC and RSP show that Cd pollution risk in the study area is relatively high, whereas As, Zn, and Cr are generally pollution-free or risk-free. In naturally developed soil in the study area, the clay mineral content and degree of soil development have significant effects on Cd migration, whereas in the cultivated layer affected by human activities, the soil pH and organic matter content are the main controlling factors. The migration ability of soil As is mainly related to soil organic matter, Fe2O3 content, and soil development degree, but organic matter has an obviously enhanced effect in the tillage layer. The main controlling factor of Zn and Cr migration in soil is pH, and the effects are more intense under the disturbance of human activities.

Propagation of desert moss Syntrichia caninervis in peat pellet: a method for rapidly obtaining large numbers of cloned gametophytes.

BACKGROUND: Syntrichia caninervis is a typical desiccation tolerant moss that is a dominant species forming biological soil crusts in the Gurbantunggut Desert. This study investigated the effect of different explants on regeneration potential by propagating them on peat pellet. RESULT: Juvenile and green leaves can regenerate secondary protonema within one week and shoots in one-half month in peat pellet. Rhizoids have a great ability to regenerate, and similar to leaf regeneration, secondary protonema is the dominant type of regenerant. The process of stem regeneration is similar to that of whole gametophytes. Stems are the most important integral body part during propagation. The whole gametophyte is the best materials for rapidly propagating gametophyte on peat pellet. CONCLUSION: This article improves the state of our current knowledge of desiccation tolerant moss cultivation, highlighting efforts to effectively obtain a large number of gametophytes through different explant parts. This work provides a useful resource for the study of S. caninervis as well as biocrust restoration.

[Factors Affecting the Translocation and Accumulation of Cadmium in a Soil-Crop System in a Typical Karst Area of Guangxi Province, China].

To understand the main factors influencing the translocation and accumulation of cadmium (Cd) in soil-crop systems in typical karst areas, 68 sets of paddy soil and rice grain samples were collected in Guangxi Province. These were used to analyze Cd concentrations and soil properties (pH, organic matter (OM) content, oxide content, and texture). Spearman's correlation coefficients and principal component analysis (PCA) were used to examine the effects of soil properties on Cd concentrations and identify the main influencing factors. The studied soils were highly enriched in iron oxide (TFe2O3), aluminum oxide (Al2O3), and manganese oxide (MnO) compared to background levels, with average concentrations of 20.2%, 19.0%, and 0.2%, respectively. However, the soils are relatively depleted in silica (SiO2), with an average concentration of 41.0%. The soils are strongly weathered and leached in study area, giving rise to rich occurrences of Fe-Mn nodules. The concentrations of TFe2O3 and MnO in the study soils were significantly correlated with soil Cd, rice seed Cd, and the Cd bioconcentration factor (BCF). The PCA analysis further showed that TFe2O3 and MnO in soils were the main factors affecting the migration and enrichment of Cd while soil pH, OM, and Al2O3 had less of an influence. Furthermore, SiO2 and soil texture indirectly affected the migration and enrichment of Cd. It is suggested that the Fe-Mn nodules effectively adsorb and immobilize Cd in the study area soils, acting as a heavy metal scavenger that reduced the biological accessibility of Cd.

Functional Aspects of Early Light-Induced Protein (ELIP) Genes from the Desiccation-Tolerant Moss Syntrichia caninervis.

The early light-induced proteins (ELIPs) are postulated to act as transient pigment-binding proteins that protect the chloroplast from photodamage caused by excessive light energy. Desert mosses such as Syntrichia caninervis, that are desiccation-tolerant and homoiochlorophyllous, are often exposed to high-light conditions when both hydrated and dry ELIP transcripts are accumulated in response to dehydration. To gain further insights into ELIP gene function in the moss S. caninervis, two ELIP cDNAs cloned from S. caninervis, ScELIP1 and ScELIP2 and both sequences were used as the basis of a transcript abundance assessment in plants exposed to high-light, UV-A, UV-B, red-light, and blue-light. ScELIPs were expressed separately in an Arabidopsis ELIP mutant Atelip. Transcript abundance for ScELIPs in gametophytes respond to each of the light treatments, in similar but not in identical ways. Ectopic expression of either ScELIPs protected PSII against photoinhibition and stabilized leaf chlorophyll content and thus partially complementing the loss of AtELIP2. Ectopic expression of ScELIPs also complements the germination phenotype of the mutant and improves protection of the photosynthetic apparatus of transgenic Arabidopsis from high-light stress. Our study extends knowledge of bryophyte photoprotection and provides further insight into the molecular mechanisms related to the function of ELIPs.

[Bioavailability, Translocation, and Accumulation Characteristic of Heavy Metals in a Soil-Crop System from a Typical Carbonate Rock Area in Guangxi, China].

This study uses 68 sets of paddy soil and rice grain samples collected from an area of carbonate rocks in Guangxi Province, China, to explore the ecological risks of heavy metals (As, Cd, Cr, Cu, Hg, Pb, Ni, and Zn) in soils from a high background area. We analyzed the concentrations of these heavy metals in soil and grain samples, and their chemical speciation in soil, and use these data to assess the associated ecological risks by means of statistics, a geo-accumulation index, bioconcentration factors (BCF), and correlation analysis. The arithmetic mean values of heavy metals concentrations in soil samples from the study area were (75.8±50.1), (1.91±1.02), (467.0±253.1), (48.5±9.8), (0.21±0.08), (76.2±28.1), (84.2±25.0), and (258.0±122.6) mg·kg-1 for As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn, respectively, which were remarkably higher than of those from other regions within Guangxi Province and China. In comparison to China's soil environmental quality standard risk screening values (GB 15618-2018), the over-standard rates of Cd, As, and Cr were 95.6%, 86.8%, and 69.1%, respectively. In comparison to risk intervention values, the over-standard rates of Cd, As, and Cr were 27.9%, 17.6%, and 5.9%, respectively. Speciation analysis on heavy metals indicated that As, Cr, Cu, Ni, Pb, and Zn were mainly found in a residual form, and accounted for>80% of the total concentrations, and had a low bioavailability. The bioactive components (F1+F2+F3) of Cd accounted for 21%, and the bioactivity of Cd was higher than other elements. The potential bioavailable components (F4+F5+F6) of Hg accounted for 44%, with low total concentrations, which are understood to have little potential ecological harm for crops. However, the over-standard rates of Pb, Cd, and Cr in rice grains were only 23.5%, 8.8%, and 2.9%, respectively. Correlation analysis showed that there was no significant correlation between the concentrations of heavy metals in soils and the corresponding rice grains. The mean BCFs of each heavy metal were <0.1, and the BCFs of Hg, Pb, As, Cr, and Ni were <0.05. Overall, we found relatively high concentrations, low activity, and low ecological risks for heavy metals in the study area. For high geological background materials such as carbonate rocks, factors such as metal speciation, biological activity, and crop over-standard rates should be taken into account along with the traditional use of the total amount of heavy metals in a soil as the evaluation standard when formulating pollution control policies.

Detection of benign proliferative lesions on vocal cords with voice handicap index.

The aim of this study was to investigate the significance and practical applicability of the voice handicap index (VHI) in the diagnosis of benign proliferative lesions of the vocal cords (BLVCs). The detection of VHI included the calculation of functional (F), physical (P) and emotional (E) domains, and the score of each domain and total score (TVH). The VHI was scored in patients with BLVCs and healthy controls. Eighty-four patients with BLVCs and 27 healthy controls were enrolled in the present study. The F, P, E and TVH scores were 10.40±7.84, 19.88±8.81, 9.39±8.49 and 39.37±21.83, respectively, in the BLVC group and 0.78±0.97, 0.85±1.06, 0.26±0.72 and 1.89±2.31, respectively, in the control group. A significant difference was found between the two groups (P<0. 01). The daily duration of speech and course of BLVCs did not correlate with the VHI score in BLVC patients (P>0.05). There was no marked difference in the VHI score between voice-consuming and non-voice consuming occupations (P>0.05) or between males and females (P>0.05). In BLVC patients, VHI may subjectively express the voice handicap, while daily duration of speech, course of BLVC, occupation and gender have no impact on VHI.

Alkaloids from Sophora flavescens Aition.

Although the quinolizidine alkaloids and flavonoids, the main active components of the traditional Chinese medicine Sophora flavescens, have been largely investigated, a new matrine alkaloid derivative 9alpha-hydroxy-7,11-dehydromatrine (1) and a rare 1,4-diazaindan-type alkaloid flavascensine (17), together with 15 known alkaloids, were isolated from S. flavescens. The structures were established on the basis of spectroscopic techniques.