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Buckwheat is a promising pseudo cereal and its cultivation history can be traced back to thousands of years ago in China. Nowadays, buckwheat is not only an ordinary crop but also a symbol of healthy life because of its rich nutritional and pharmacological properties. In this research, the current suitable areas of 19 wild buckwheat species were analyzed by the MaxEnt model, which proved that southwestern China was the diversity center of buckwheat. Their morphological characteristics and geographical distribution were analyzed for the first time. In addition, it was found that the change of buckwheat cultivation in three periods might be related to the green revolution of main crops and national policies. Meanwhile, the Sustainable Yield Index (SYI) value of buckwheat in China was the lowest from 1959 to 2016. Through the MaxEnt model, the potentially suitable areas of wild buckwheat would contract while cultivated buckwheat would expand under climate change. Accordingly, the diversity of wild buckwheat will decrease. Therefore, it is necessary to protect buckwheat resources as much as possible to strengthen the development and utilization of buckwheat resources. Moreover, the promotion of buckwheat diversity will be an important trade-off between food security, population growth, and land use under climate change.

A novel actinomycete strain, designated strain QMT-12

Lotus corniculatus is exposed to many forms of biotic stresses, including pathogen and herbivore attack. Jasmonates (JAs) are lipid-based signaling molecules that play important roles in mediating secondary metabolite biosynthesis and plant defense responses. Here, we first characterized LcJAZ1 and LcMYC2, two key components of the JAs signaling pathway. Both LcJAZ1 and LcMYC2 mRNA accumulated quickly in response to JAs. Subcellular localization analysis showed that both LcJAZ1 and LcMYC2 are nuclear proteins. In addition, LcJAZ1 protein was degraded after JAs treatment, but LcMYC2 is not. Yeast two-hybrid, pull-down, and bimolecular fluorescence complementation assays demonstrated that LcJAZ1 interacts with LcMYC2 and AtMYC2. In addition, LcMYC2 interacts with AtJAZ1. The Arabidopsis protoplast transactivation assay revealed that LcJAZ1 acts as repressors of LcMYC2- or AtMYC2-mediated transcription. Our data indicate that the conserved JAZ-MYC complex contributes to the modulation of the JAs signaling pathway.

Terpenoids are the largest class of secondary metabolites with indispensable functions for plant growth and development. Maize is a very important cereal crop that is grown widely throughout the world. However, many genes and gene family members involved in terpenoid biosynthesis are still unknown in maize. Through systematic analysis, 43 isoprenoid diphosphate biosynthesis-related genes were identified. These genes are members of 15 families, which encode all of the enzymes involved in the biosynthesis of the universal isoprene precursor isopentenyl diphosphate and its isomer dimethylallyl diphosphate. Using a comprehensive approach, the intron/exon structures and their responses to jasmonic acid treatment were investigated. Meta-analysis revealed that some of the genes show differential, organ-specific expression patterns in the 60 different developmental stages of maize. It was found that all 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway genes present the same expression patterns which were highly expressed in leaves and showed the lowest expression in seed. All investigated MEP pathway enzymes are localized in plastids by transient assay using Arabidopsis cells. In addition, some of the enzymes, such as acetyl-CoA C-acetyltransferase, hydroxymethylglutaryl-CoA reductase, and geranylgeranyl diphosphate synthase, are encoded by multiple gene members with different expression patterns, and both homomeric and heteromeric geranyl diphosphate synthases exist in maize. The results reveal the complexity of terpenoid biosynthesis and the existence of metabolic channels for diverse terpenoids in maize.

Buckwheat is a nutritional and economically crop belonging to Polygonaceae, Fagopyrum. To better understand the mutation patterns and evolution trend in the chloroplast (cp) genome of buckwheat, and found sufficient number of variable regions to explore the phylogenetic relationships of this genus, two complete cp genomes of buckwheat including Fagopyrum dibotrys (F. dibotrys) and Fagopyrum luojishanense (F. luojishanense) were sequenced, and other two Fagopyrum cp genomes were used for comparative analysis. After morphological analysis, the main difference among these buckwheat were height, leaf shape, seeds and flower type. F. luojishanense was distinguishable from the cultivated species easily. Although the F. dibotrys and two cultivated species has some similarity, they different in habit and component contents. The cp genome of F. dibotrys was 159,320 bp while the F. luojishanense was 159,265 bp. 48 and 61 SSRs were found in F. dibotrys and F. luojishanense respectively. Meanwhile, 10 highly variable regions among these buckwheat species were located precisely. The phylogenetic relationships among four Fagopyrum species based on complete cp genomes was showed. The results suggested that F. dibotrys is more closely related to Fagopyrum tataricum. These data provided valuable genetic information for Fagopyrum species identification, taxonomy, phylogenetic study and molecular breeding.

The APETALA2/ethylene-responsive element binding factors (AP2/ERF) play central roles in the stress response in plants. In this study, we identified and isolated a novel salt stress-related gene, LcERF080, that encodes an AP2/ERF protein in Lotus corniculatus cultivar Leo. LcERF080 was classified into the B-4 group of the ERF subfamily based on multiple sequence alignment and phylogenetic characterization. Expression of LcERF080 was strongly induced by salt, abscisic acid, 1-aminocyclopropane-1-carboxylic acid, methyl jasmonate, and salicylic acid stresses. Subcellular localization assay confirmed that LcERF080 is a nuclear protein. LcERF080 overexpression in Arabidopsis resulted in pleiotropic phenotypes with a higher seed germination rate and transgenic plants with enhanced tolerance to salt stress. Further, under stress conditions, the transgenic lines exhibited elevated levels of soluble sugars and proline as well as relative moisture contents but a lower malondialdehyde content than in control plants. The expression levels of hyperosmotic salinity response genes COR15A, RD22, and P5CS1 were found to be elevated in the LcERF080-overexpressing Arabidopsis plants compared to the wild-type plants. These results reveal that LcERF080 is involved in the responses of plants to salt stress.

Lotus corniculatus is used in agriculture as a main forage plant. Members of the Apetala2/ethylene response factor (AP2/ERF) family play important roles in regulating gene expression in response to many forms of stress, including drought and salt. Here, starting from database of the L. corniculatus var. japonicus genome, we identified 127 AP2/ERF genes by insilico cloning method. The phylogeny, gene structures, and putative conserved motifs in L. corniculatus var. japonicus ERF proteins were analyzed. Based on the number of AP2/ERF domains and the function of the genes, 127 AP2/ERF genes from L. corniculatus var. japonicus were classified into five subfamilies named the AP2, dehydration-responsive element binding factor (DREB), ERF, RAV, and a soloist. Outside the AP2/ERF domain, many L. corniculatus var. japonicus-specific conserved motifs were detected. Expression profile analysis of AP2/ERF genes by quantitative real-time PCR revealed that 19 LcERF genes, including LcERF054 (KJ004728), were significantly induced by salt stress. The results showed that the LcERF054 gene encodes a nuclear transcription activator. Overexpression of LcERF054 in Arabidopsis enhanced the tolerances to salt stress, showed higher germination ratio of seeds, and had elevated levels of relative moisture contents, soluble sugars, proline, and lower levels of malondialdehyde under stress conditions compared to wild-type plants. The expression of hyperosmotic salinity response genes COR15A, LEA4-5, P5CS1, and RD29A was found to be elevated in the LcERF054-overexpressing Arabidopsis plants compared to wild type. These results revealed that the LcERF genes play important roles in L. corniculatus cv Leo under salt stress and that LcERFs are attractive engineering targets in applied efforts to improve abiotic stress tolerances in L. corniculatus cv Leo or other crops.

Maize is a cereal crop that is grown widely throughout the world in a range of agro-ecological environments. Trehalose is a nonreducing disaccharide of glucose that has been associated with tolerance to different stress conditions, including salt and drought. Bioinformatic analysis of genes involved in trehalose biosynthesis and degradation in maize has not been reported to date. Through systematic analysis, 1 degradation-related and 36 trehalose biosynthesis-related genes were identified. The conserved domains and phylogenetic relationships among the deduced maize proteins and their homologs, isolated from other plant species such as Arabidopsis and rice, were revealed. Using a comprehensive approach, the intron/exon structures and expression patterns of all identified genes and their responses to salt stress, jasmonic acid, and abscisic acid treatment were analyzed. Microarray data demonstrated that some of the genes show differential, organ-specific expression patterns in the 60 different developmental stages of maize. It was discovered that some of the key enzymes such as hexokinase, trehalose-6-phosphate synthase, and trehalose-6-phosphate phosphatase are encoded by multiple gene members with different expression patterns. The results highlight the complexity of trehalose metabolism and provide useful information for improving maize stress tolerance through genetic engineering.

Nicotianamine (NA) is an important divalent metal chelator and the main precursor of phytosiderophores. NA is synthesized from S-adenosylmethionine in a process catalyzed by nicotianamine synthase (NAS). In this study, a set of structural and phylogenetic analyses have been applied to identify the maize NAS genes based on the maize genome sequence release. Ten maize NAS genes have been mapped; seven of them have not been reported to date. Phylogenetic analysis and expression pattern from microarray data led to their classification into two different orthologous groups. C-terminal fusion of ZmNAS3 with GFP was found in the cytoplasm of Arabidopsis leaf protoplast. Expression analysis by reverse transcription polymerase chain reaction revealed ZmNAS genes are responsive to heavy metal ions (Ni, Fe, Cu, Mn, Zn, and Cd), and all 10 ZmNAS genes were only observed in the root tissue except of ZmNAS6. The promoter of ZmNAS genes was analyzed for the presence of different cis-element response to all kinds of phytohormones and environment stresses. We found that the ZmNAS gene expression of maize seedlings was regulated by jasmonic acid, abscisic acid, and salicylic acid. Microarray data demonstrated that the ZmNAS genes show differential, organ-specific expression patterns in the maize developmental steps. The integrated comparative analysis can improve our current view of ZmNAS genes and facilitate the functional characterization of individual members.

Maize (Zea mays ssp. mays L.) is an important model organism for fundamental research in the agro-biotechnology field. Aldehydes were generated in response to a suite of environmental stresses that perturb metabolism including salinity, dehydration, desiccation, and cold and heat shock. Many biologically important aldehydes are metabolized by the superfamily of NAD(P)(+)-dependent aldehyde dehydrogenases. Here, starting from the database of Z. mays, we identified 28 aldehyde dehydrogenase (ALDH) genes and 48 transcripts by the in silico cloning method using the ALDH-conserved domain amino acid sequence of Arabidopsis and rice as a probe. Phylogenetic analysis shows that all 28 members of the ALDH gene families were classified to ten distinct subfamilies. Microarray data and quantitative real-time PCR analysis reveal that ZmALDH9, ZmALDH13, and ZmALDH17 genes involve the function of drought stress, acid tolerance, and pathogens infection. These results suggested that these three ZmALDH genes might be potentially useful in maize genetic improvement.

A novel DREB (dehydration-responsive element binding) gene, designated PeDREB2a, was isolated from the desert-grown tree, Populus euphratica Oliv. PeDREB2a is classified into the A-5 group of DREB subfamily based on multiple sequence alignment and phylogenetic characterization. Using semi-quantitative RT-PCR, we found that the PeDREB2a was greatly induced by drought, NaCl, low temperature, 1-naphthaleneacetic acid (NAA), 6-benzyl aminopurine (6-BA) and gibberellic acid (GA3) treatments in P. euphratica seedling. Yeast transactivity assay demonstrated that PeDREB2a gene encodes a transcription activator. Overexpression of PeDREB2a under the stress-inducible rd29A promotor in transgenic Arabidopsis and Lotus corniculatus forage plants resulted in enhanced tolerance to salt and drought stresses. The PeDREB2a overexpressing Arabidopsis lines showed higher root length and plant height and had elevated levels of soluble sugars and lower levels of malondialdehyde under stress conditions compared to control plants. The results revealed that PeDREB2a play an essential role as a DREB transcription factor in regulation of stress-responsive signaling in P. euphratica.

Significant progress has been made in recent years in enhancing the ability of plants to tolerate, remove, and degrade pollutants. Plant root remediation of contaminated soils and groundwater shows great potential for future development due to its environmental compatibility and cost-effectiveness. Hairy roots are disease manifestations developed by plants that are wounded and infected by Agrobacterium rhizogenes. The application of transgenic hairy roots in phytoremediation has been suggested mainly because of their biochemical resemblance to the roots of the plant from which they are derived. The application of genetic engineering has greatly augmented removal rates of hazardous pollutants. In addition, the rhizospheric bacteria that live on or around plant hairy roots also lead to improved tolerance to normally phytotoxic chemicals and increased removal of pollutants. This paper provides a broad overview of the evidence supporting the suitability and prospects of hairy roots in phytoremediation of organic pollutants and heavy metals.

Alfalfa is excellent perennial legume forage for its extensive ecological adaptability, high nutrition value, palatability and biological nitrogen fixation. It plays a very important role in the agriculture, animal husbandry and ecological construction. It is cultivated in all continents. With the development of modern plant breeding and genetic engineering techniques, a large amount of work has been carried out on alfalfa. Here we summarize the recent research advances in genetic engineering of alfalfa breeding, including transformation, quality improvement, stress resistance and as a bioreactor. The review article can enables us to understand the research method, direction and achievements of genetic engineering technology of Alfalfa.

Lotus corniculatus is used in agriculture as a main forage plant. Apetala2/Ethylene response factor (AP2/ERF) family plays an important role in regulating the gene expression in response to many forms of stress. Here, we identified 127 AP2/ERF genes by in silico cloning method. The phylogeny , gene structures, and putative conserved motifs in L. japonicus ERF proteins were analyzed. Expression profile analysis of AP2/ERF genes by quantitative real-time PCR revealed that 19 LcERF genes, including B-2 group LcERF054 and B-4 group LcERF080, were significantly induced by salt stress . Overexpression of LcERF054 or LcERF080 in Arabidopsis enhanced the tolerances to salt stress, showed elevated levels of relative moisture contents, soluble sugars, proline , and lower levels of malondialdehyde under stress conditions compared to wild-type. The expression of hyperosmotic salinity response genes COR15A, P5CS1, and so on were found to be elevated in the LcERF054 or LcERF080 overexpressing Arabidopsis plants compared to wild-type. These results revealed that LcERF genes play important roles in L. corniculatus cv Leo under salt stress and that LcERFs are attractive engineering targets in applied efforts to improve abiotic stress tolerances in L. corniculatus or other crops.

Fagopyrum longzhoushanense (Polygonaceae) is here described and illustrated as a new species from Longzhoushan (Puge County, Sichuan Province, China) on the basis of morphological, caryological and molecular analyses. The new species is morphologically similar to F. gracilipes from which it differs in having densely pubescent or villose parts, thicker and red-brown stems and branches, dense nodes, rugulose and small postulate leaf blade on the surface, stamens longer than pistils. Another similar species is F. densovillosum from which F. longzhoushanense differs by the winged and greater achenes, and the stamens which are longer than the pistils. From the caryioogical point of view F. longzhoushanense is a diploid species with 2n=2x=16 and a karyotype of 16 metacentric chromosomes, while F. gracilipes is tetraploid, 4n=4x=32, with a karyotype of 30 metacentric and 2 submetacentric chromosomes. The results of molecular marker analysis (matK) also highlight the distinction of the new species.

The rutin degrading enzyme (RDE) was isolated and purified from tartary buckwheat seeds. The RDE was purified about 11.34-fold and its final yield was 3.5%, which was very low, due to our purification strategy of giving priority to purity over yield. The RDE molecular weight was estimated to be about 60 kDa. When rutin was used as substrate, an optimal enzyme activity was seen at around pH 5.0 and 40 °C. Strains isolation strategy characterized by the use of rutin as sole carbon source in enrichment cultures was used to isolate RDE-producing strains. Then the active strains were identified by morphology characterization and 18s rDNA-ITS (Internal Transcribed Spacer) gene sequencing. Three isolates coded as B3, W2, Y2 were successfully isolated from fusty Fagopyrum tataricum flour cultures. Strain B3 possessed the highest unit activity among these three strains, and its total activity reached up to 171.0 Unit. The active isolate (B3) could be assigned to Penicillium farinosum. When the Penicillium farinosum strains were added to tartary buckwheat flour cultures at pH 5.0, 30 °C after 5 days fermentation, the quercetin production raised up to 1.78 mg/l, almost 5.1 times higher than the fermentation without the above active strains. Hence, a new approach was available to utilize microorganism-aided fermentation for effective quercetin extraction from Fagopyrum tataricum seeds.

This study was aimed at investigating the purification and identification of serine protease inhibitors, F. tataricum trypsin inhibitor (FtTI) from tartary buckwheat (Fagopyrum tataricum) seeds. The FtTI was isolated by anion exchange chromatography, affinity chromatography, and centrifugal ultrafiltration. Under reducing and nonreducing conditions, an SDS-PAGE analysis showed that the isolated protein consists of a single polypeptide chain with a molecular mass of approximately 14 kDa. The two isoforms of FtTI were confirmed by the mass spectrometric profile where the two peaks corresponded to 11.487 and 13.838 kDa. The complete amino acid sequence of FtTI has been established by automatic Edman degradation and mass spectrometry. The molecule of FtTI consists of 86 amino acid residues containing two disulfide bonds which connect Cys8 to Cys65 and Cys49 to Cys58. The active site of FtTI contains an Asp66-Arg67 bond. The Ki value was calculated using the equation for slow tight binding inhibition which was 1.6 nM for trypsin. FtTI retained its inhibitory activity over a wide range of pH (3-10) and temperature (20-80 °C). FtTI can be rapidly inactivated by the combination of high temperature and high pressure. An analysis of the amino acid sequence suggests that FtTI is a member of the protease inhibitor Ι family. Furthermore, FtTI exhibited a strong inhibitory activity against phytopathogenic fungi.