Genome-wide identification, structural characterization and gene expression analysis of the WRKY transcription factor family in pea (Pisum sativum L.).

BACKGROUND: The WRKY gene family is one of the largest families of transcription factors in higher plants, and WRKY transcription factors play important roles in plant growth and development as well as in response to abiotic stresses; however, the WRKY gene family in pea has not been systematically reported. RESULTS: In this study, 89 pea WRKY genes were identified and named according to the random distribution of PsWRKY genes on seven chromosomes. The gene family was found to have nine pairs of tandem duplicates and 19 pairs of segment duplicates. Phylogenetic analyses of the PsWRKY and 60 Arabidopsis WRKY proteins were performed to determine their homology, and the PsWRKYs were classified into seven subfamilies. Analysis of the physicochemical properties, motif composition, and gene structure of pea WRKYs revealed significant differences in the physicochemical properties within the PsWRKY family; however, their gene structure and protein-conserved motifs were highly conserved among the subfamilies. To further investigate the evolutionary relationships of the PsWRKY family, we constructed comparative syntenic maps of pea with representative monocotyledonous and dicotyledonous plants and found that it was most recently homologous to the dicotyledonous WRKY gene families. Cis-acting element analysis of PsWRKY genes revealed that this gene family can respond to hormones, such as abscisic acid (ABA), indole-3-acetic acid (IAA), gibberellin (GA), methyl jasmonate (MeJA), and salicylic acid (SA). Further analysis of the expression of 14 PsWRKY genes from different subfamilies in different tissues and fruit developmental stages, as well as under five different hormone treatments, revealed differences in their expression patterns in the different tissues and fruit developmental stages, as well as under hormone treatments, suggesting that PsWRKY genes may have different physiological functions and respond to hormones. CONCLUSIONS: In this study, we systematically identified WRKY genes in pea for the first time and further investigated their physicochemical properties, evolution, and expression patterns, providing a theoretical basis for future studies on the functional characterization of pea WRKY genes during plant growth and development.

Genome-wide identification, evolution, and role of SPL gene family in beet (Beta vulgaris L.) under cold stress.

BACKGROUND: SPL transcription factors play vital roles in regulating plant growth, development, and abiotic stress responses. Sugar beet (Beta vulgaris L.), one of the world's main sugar-producing crops, is a major source of edible and industrial sugars for humans. Although the SPL gene family has been extensively identified in other species, no reports on the SPL gene family in sugar beet are available. RESULTS: Eight BvSPL genes were identified at the whole-genome level and were renamed based on their positions on the chromosome. The gene structure, SBP domain sequences, and phylogenetic relationship with Arabidopsis were analyzed for the sugar beet SPL gene family. The eight BvSPL genes were divided into six groups (II, IV, V, VI, VII, and VIII). Of the BvSPL genes, no tandem duplication events were found, but one pair of segmental duplications was present. Multiple cis-regulatory elements related to growth and development were identified in the 2000-bp region upstream of the BvSPL gene start codon (ATG). Using quantitative real-time polymerase chain reaction (qRT-PCR), the expression profiles of the eight BvSPL genes were examined under eight types of abiotic stress and during the maturation stage. BvSPL transcription factors played a vital role in abiotic stress, with BvSPL3 and BvSPL6 being particularly noteworthy. CONCLUSION: Eight sugar beet SPL genes were identified at the whole-genome level. Phylogenetic trees, gene structures, gene duplication events, and expression profiles were investigated. The qRT-PCR analysis indicated that BvSPLs play a substantial role in the growth and development of sugar beet, potentially participating in the regulation of root expansion and sugar accumulation.

Genome-wide identification, evolution and expression pattern analysis of the GATA gene family in Sorghum bicolor.

The GATA family of transcription factors is zinc finger DNA binding proteins involved in a variety of biological processes, including plant growth and development and response to biotic/abiotic stresses, and thus play an essential role in plant response to environmental changes. However, the GATA gene family of Sorghum (SbGATA) has not been systematically analyzed and reported yet. Herein, we used a variety of bioinformatics methods and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to explore the evolution and function of the 33 SbGATA genes identified. These SbGATA genes, distributed on 10 chromosomes, are classified into four subfamilies (I-IV) containing one pair of tandem duplications and nine pairs of segment duplications, which are more closely related to the monocot Brachypodium distachyon and Oryza sativa GATA genes. The physicochemical properties of the SbGATAs are significantly different among the subfamilies, while the protein structure and conserved protein motifs are highly conserved in the subfamilies. In addition, the transcription of SbGATAs is tissue-specific during Sorghum growth and development, which allows for functional diversity in response to stress and hormones. Collectively, our study lays a theoretical foundation for an in-depth analysis of the functions, mechanisms and evolutionary relationships of SbGATA during plant growth and development.

bHLH transcription factor family identification, phylogeny, and its response to abiotic stress in Chenopodium quinoa.

The second-largest transcription factor superfamily in plants is that of the basic helix-loop-helix (bHLH) family, which plays an important complex physiological role in plant growth, tissue development, and environmental adaptation. Systematic research on the Chenopodium quinoa bHLH family will enable a better understanding of this species. Herein, authors used a variety of bioinformatics methods and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to explore the evolution and function of the 218 CqbHLH genes identified. A total of 218 CqbHLH transcription factor genes were identified in the whole genome, located on 18 chromosomes. A phylogenetic tree was constructed using the CqbHLH and AtbHLH proteins to determine their homology, and the members were divided into 20 subgroups and one unclustered gene. Authors also analyzed 218 CqbHLH genes, conservative motifs, chromosome diffusion, and gene replication. The author constructed one Neighbor-Joining (NJ) tree and a collinearity analysis map of the bHLH family in C. quinoa and six other plant species to study the evolutionary relationship and homology among multiple species. In addition, the expression levels of 20 CqbHLH members from different subgroups in various tissues, different fruit developmental stages, and six abiotic stresses were analyzed. Authors identified 218 CqbHLH genes and studied their biological functions, providing a basis for better understanding and further studying the bHLH family in quinoa.

FtbZIP12 Positively Regulates Responses to Osmotic Stress in Tartary Buckwheat.

ABFs play a key role in regulating plant osmotic stress. However, in Tartary buckwheat, data on the role of ABF genes in osmotic stress remain limited and its associated mechanism in osmoregulation remain nebulous. Herein, a novel ABF family in Tartary buckwheat, FtbZIP12, was cloned and characterized. FtbZIP12 is a transcriptional activator located in the nucleus; its expression is induced by NaCl, mannitol, and abscisic acid (ABA). Atopic expression of FtbZIP12 in Arabidopsis promoted seed germination, reduced damage to primary roots, and improved the tolerance of seedlings to osmotic stress. The quantitative realtime polymerase chain reaction (RT-qPCR) results showed that the expressions of the typical genes related to stress, the SOS pathway, and the proline synthesis pathway in Arabidopsis were significantly (p < 0.05) upregulated under osmotic stress. FtbZIP12 improved the osmotic pressure resistance by reducing the damage caused by reactive oxygen species to plants and maintained plant homeostasis by upregulating the expression of genes related to stress, osmotic regulation, and ion homeostasis. This study identified a key candidate gene for understanding the mechanism underlying osmotic-stress-regulated function in Tartary buckwheat, thereby providing a theoretical basis for improving its yield and quality.

Genome-Wide Identification, Evolution, and Expression Pattern Analysis of the GATA Gene Family in Tartary Buckwheat (Fagopyrum tataricum).

GATA is a transcription factor that exerts a vital function in plant growth and development, physiological metabolism, and environmental responses. However, the GATA gene family has rarely been studied in Tartary buckwheat since the completion of its genome. This study used bioinformatics methods to identify GATA genes of Tartary buckwheat and to analyze their subfamily classification, structural composition, and developmental evolution, as well as to discuss the expression patterns of FtGATA genes in different subfamilies. The twenty-eight identified FtGATA genes in the Tartary buckwheat genome were divided into four subfamilies and distributed on eight chromosomes. One pair of tandem repeat genes and eight pairs of fragments were found in chromosome mapping. Spatiotemporal expression patterns of eight FtGATA genes in different subfamilies indicated that the FtGATA gene family has regulatory roles in tissue specificity, fruit development, abiotic stress, and hormonal responses. This study creates a theoretical and scientific foundation for further research on the evolutionary relationship and biological function of FtGATA.

Physiological and Biochemical Regulation Mechanism of Exogenous Hydrogen Peroxide in Alleviating NaCl Stress Toxicity in Tartary Buckwheat (Fagopyrum tataricum (L.) Gaertn).

We aimed to elucidate the physiological and biochemical mechanism by which exogenous hydrogen peroxide (H2O2) alleviates salt stress toxicity in Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn). Tartary buckwheat "Chuanqiao-2" under 150 mmol·L-1 salt (NaCl) stress was treated with 5 or 10 mmol·L-1 H2O2, and seedling growth, physiology and biochemistry, and related gene expression were studied. Treatment with 5 mmol·L-1 H2O2 significantly increased plant height (PH), fresh and dry weights of shoots (SFWs/SDWs) and roots (RFWs/RDWs), leaf length (LL) and area (LA), and relative water content (LRWC); increased chlorophyll a (Chl a) and b (Chl b) contents; improved fluorescence parameters; enhanced antioxidant enzyme activity and content; and reduced malondialdehyde (MDA) content. Expressions of all stress-related and enzyme-related genes were up-regulated. The F3'H gene (flavonoid synthesis pathway) exhibited similar up-regulation under 10 mmol·L-1 H2O2 treatment. Correlation and principal component analyses showed that 5 mmol·L-1 H2O2 could significantly alleviate the toxic effect of salt stress on Tartary buckwheat. Our results show that exogenous 5 mmol·L-1 H2O2 can alleviate the inhibitory or toxic effects of 150 mmol·L-1 NaCl stress on Tartary buckwheat by promoting growth, enhancing photosynthesis, improving enzymatic reactions, reducing membrane lipid peroxidation, and inducing the expression of related genes.

Roles of Arbuscular mycorrhizal Fungi as a Biocontrol Agent in the Control of Plant Diseases.

Arbuscularmycorrhizal fungi (AMF) are a class of beneficial microorganisms that are widely distributed in soil ecosystems and can form symbionts with 80% of terrestrial higher plants, and improve the nutritional status of plants. The use of AMF as a biocontrol method to antagonize soil-borne pathogens has received increasing interest from phytopathologists and ecologists. In this paper, the mechanisms of resistance to diseases induced by AMF and the application of AMF to plant fungal, bacterial, and nematode diseases have been summarized. This study aimed to enhance the potential use of AMF as a biological control method to prevent plant diseases in the future. Root morphological alteration characteristics were explained, including the influence of AMF on root structure, function, and the regulation of AMF via secondary metabolites. AMF can improve the rhizosphere environment by influencing the physical and chemical proprieties of soil, enhancing the growth of other beneficial microorganisms, and by competing with pathogenic microorganisms. Two microorganism types may compete for the same invasive sites in root systems and regulate nutrition distribution. AMF can induce the host plant to form defense systems, including improving phytohormone concentrations, inducing signal substrate production, gene expression regulation, and enhancing protein production.

First Report of Botryosphaeria dothidea Causing Gray Mold on Tartary Buckwheat in Southwest China.

Tartary buckwheat (Fagopyrum tataricum, Polygonaceae) is an annual plant originating in Southwest China. It has a short growth cycle, barren soil tolerance, and strong stress resistance (Zhang et al. 2021). Because of its high content of proteins, starch, trace elements, phenols, and dietary fiber, Tartary buckwheat is beneficial to the human body and hence has received widespread attention (Joshi et al. 2019; Dc ja, B, et al. 2020). In the period from September to November 2020, a diseased plant infected with gray mold was found among M2 generation plants treated using ethyl methanesulfonate (EMS) in a location with potted Tartary buckwheat plants in Huaxi District, Guiyang City, Guizhou Province, China. The diseased plant started to show symptoms during the initial flowering stage; water-soaked spots appeared at first, that the spots increased in size and turned into light brown patches, with the leaf edges scorched brown. In severe cases, the leaves turned yellow, the diseased spots became dry, and finally the leaves necrotic (Figure 1A). Among the leaves that showed disease symptoms, severely susceptible leaves were selected; a piece of tissue (2×2 mm) was removed at the junction of the diseased and healthy tissues. The tissue was then soaked in 75% ethanol for 2 to 3 s, transferred to 1% sodium hypochlorite solution and soaked for 3 min, rinsed three times with sterile water, and placed on sterilized filter paper to dry. Sterile tweezers were used to transfer the tissue blocks to Potato Dextrose Agar medium (Bio-Rad Ltd. Com, USA) containing a Streptomyces-Penicillium mixture (100 µg/mL), and they were incubated on this medium for 7 to 10 days at 25°C and 70% humidity under 16 h light and 8 h dark conditions. The colonies were white at the early stages, with developed aerial hyphae; subsequently, they gradually turned gray-green (Figure 1B). In the later stages, the back of the colony was black and piles of conidia could be seen (Figure 1C). The conidia are scattered, which were colorless and transparent, fusiform or fusiform, with a size of 8.02-11.13 µm×2.06-3.22 µm (average=9.51 µm×2.69 µm, n=50) (Figure 1D). Based on their morphological characteristics, These cultural and morphological characteristics were consistent with the descriptions of as B. dothidea (Fan et al. 2021). The ITS1/ITS4 (Mills et al. 1992), Bt-2a/Bt-2b primers (Glass and Donaldson 1995), and EF1-728F/EF1-986R (Slippers et al. 2004) were amplified and sequenced to analyze the ITS region, ß-tubulin genes translation elongation factor 1-α (TEF1-α), and translation elongation factor 1-α (TEF1-α), respectively. According to BLAST search in GenBank, the sequences of ITS (MZ326853), TUB2 (MZ399162) and TEF1-α (MZ399163) had 99.40%, 100% and 100% similarity to sequences NR111146.1, AY236927.1, and AY236898.1 of B. dothidea ex-type strain CMW8000, respectively. The three nucleotide sequences were concatenated together, and MEGA-X (with the neighbor-joining method) with 1,000 bootstraps was used to construct a phylogenetic tree. The results showed that our isolate was closely related to B. dothidea (Figure 2). Healthy Tartary buckwheat from the M2 generation was used for the pathogenicity test. Disinfect with 75% alcohol and 1×105 mL-1 of spore suspension was sprayed on the leaves. Each treatment included three plants, and it was repeated three times with sterile water as control. The treatments were kept in a houseat25°C for 24 h, then transferred it to the natural environment of 22℃ to 28℃,and sterile water was sprayed every morning and evening to keep the leaves moist. After 10 days, the symptoms seen in the field appeared on the treated plants (Figure 1E), but the control plants did not show any symptoms (Figure 1F). The diseased parts of the leaves were isolated and cultured again, and the isolates were consistent with the original inoculum. Thus, the study conformed to Koch's postulates. B. dothidea is a fungus with no host preference in the genus Botryosphaeria (Botryosphaeriaceae, Botryosphaeriales). It can cause canker, leaf spots, trunk diseases, fruit rot and die-back of many important wood plants all over the world (Marsberg et al.2017). Recently, it was reported that B. dothidea caused soybean canker in China (Chen et al.2021), but there have been no reports of B. dothidea causing Tartary buckwheat gray mold. To the best of our knowledge, this is the first report of B. dothidea causing gray mold on Tartary buckwheat. This finding will provide a basis for the prevention and treatment of Tartary buckwheat gray mold.

Application of Fourier transform near-infrared spectroscopy combined with GC in rapid and simultaneous determination of essential components in Amomum villosum.

A method is described using rapid and sensitive Fourier transform near-infrared spectroscopy combined with Gas Chromatograpy internal standard method detection for the simultaneous identification and determination of three bioactive compounds in Amomum villosum samples. Partial least squares regression is selected as the analysis type and multiplicative scatter correction, second derivative, and SNV were adopted for the spectral pretreatment. The correlation coefficients (R) of the calibration models were above 0.95 and the root mean square error of predictions were under 0.8. The developed models were applied to unknown samples with satisfantory results. The established method was validated and can be applied to the intrinsic quality control of Amomum villosum.

Development of a comprehensive quality control method for the quantitative analysis of volatiles and lignans in Magnolia biondii Pamp. by near infrared spectroscopy.

The quality of drug is vital to its curative effect, thus it is important to develop a comprehensive quality control method for commonly used drugs. In this study, we developed a Gas chromatography-mass spectrometry separation method for the qualitative and quantitative analysis of volatiles, together with a High-performance liquid chromatography-mass spectrometry separation method for lignans in Magnolia biondii Pamp.. 79 volatiles and 11 lignans were identified via comparing their chromatographic behavior and mass spectra data with those in the literature. The methods were then used to determine the contents of volatiles (1, 8-cineole, d-Limonene, α-terpineol, linalool, L-camphor brain and bornyl acetate) and lignans (epieudesmin, magnolin, epi-magnolin A and fargesin) in Magnolia biondii Pamp.. Subsequently, 13 qualitative models including volatiles (1, 8-cineole, d-Limonene, α-terpineol, linalool, L-camphor brain and bornyl acetate), water-soluble extractive, lignans (pinoresinol dimethyl ether, magnolin, epi-magnolin A and fargesin) and moisture were developed by Near-Infrared Spectroscopy based on partial least square regression herein. The reference values were obtained by High-performance liquid chromatography, Gas chromatography and etc., while the predicted values were attained from the NIR spectrum. Compared with the traditional detection methods, NIR technique methodology significantly improved the ability to evaluate the quality of Magnolia biondii Pamp., which had the advantages of convenience, celerity, highly efficiency, low cost, no harm to samples, no reagent consumption, and no pollution to the environment. Moreover, the systematic analysis method combined pharmaceutical analysis with pharmacochemistry was proposed to prepare volatiles, water-soluble extractive and lignans parts from the same sample. This way could extract more index components to be beneficial in the quality control of Magnolia biondii Pamp. roundly.

Coix lacryma-jobi chymotrypsin inhibitor displays antifungal activity.

A novel chymotrypsin inhibitor, named ClCI, was purified from coix seed (Coix lacryma-jobi L.) by aqueous two-phase extraction, chymotrypsin-Sepharose 4B affinity chromatography and centrifugal ultrafiltration. ClCI was a 7.9 kDa competitive inhibitor with pI 6.54. The inhibition constants (Ki) for bovine pancreatic chymotrypsin and bacterial subtilisin were 1.27 × 10-10 M and 1.57 × 10-9 M respectively. ClCI had no inhibitory activity against bovine trypsin and porcine elastase. ClCI had wide pH stability and good heat resistance. It can maintain >90% inhibition activity against chymotrypsin at 20-80 °C for 1 h. The primary structure of ClCI was highly similar (57%-92%) to those of several inhibitors belonging to the Gramineae crop potato protease inhibitor- I superfamily and showed the typical sequence motif of the protease inhibitor of the seed storage protein group. ClCI (12.5 mg) inhibited mycelial growth of the phytopathogenic fungi Mycosphaerella melonis, Helminthosporium turcicum, Alternaria solani, Phytophthora capsici, Isariopsis griseola, and Colletotrichum gloeosporioides, and caused 89% inhibition of the proteases from spore germination of plant-pathogenic fungi. The results of the present study indicate that ClCI had biotechnological potential as an alternative agent to combat the important phytopathogenic fungi.

Jasmonic Acid Signaling Pathway in Plants.

Jasmonic acid (JA) and its precursors and dervatives, referred as jasmonates (JAs) are important molecules in the regulation of many physiological processes in plant growth and development, and especially the mediation of plant responses to biotic and abiotic stresses. JAs biosynthesis, perception, transport, signal transduction and action have been extensively investigated. In this review, we will discuss the initiation of JA signaling with a focus on environmental signal perception and transduction, JA biosynthesis and metabolism, transport of signaling molecules (local transmission, vascular bundle transmission, and airborne transportation), and biological function (JA signal receptors, regulated transcription factors, and biological processes involved).