Morphological characterization and transcriptome analysis of rolled and narrow leaf mutant in soybean.

BACKGROUND: In plants, the leaf functions as a solar panel, where photosynthesis converts carbon dioxide and water into carbohydrates and oxygen. In soybean, leaf type traits, including leaf shape, leaf area, leaf width, and leaf width so on, are considered to be associated with yield. In this study, we performed morphological characterization, transcriptome analysis, and endogenous hormone analysis of a rolled and narrow leaf mutant line (rl) in soybean. RESULTS: Compared with wild type HX3, mutant line rl showed rolled and narrower leaflet, and smaller leaf, meanwhile rl also performed narrower pod and narrower seed. Anatomical analysis of leaflet demonstrated that cell area of upper epidermis was bigger than the cell area of lower epidermis in rl, which may lead rolled and narrow leaf. Transcriptome analysis revealed that several cytokinin oxidase/dehydrogenase (CKX) genes (Glyma.06G028900, Glyma.09G225400, Glyma.13G104700, Glyma.14G099000, and Glyma.17G054500) were up-regulation dramatically, which may cause lower cytokinin level in rl. Endogenous hormone analysis verified that cytokinin content of rl was lower. Hormone treatment results indicated that 6-BA rescued rolled leaf enough, rescued partly narrow leaf. And after 6-BA treatment, the cell area was similar between upper epidermis and lower epidermis in rl. Although IAA content and ABA content were reduced in rl, but exogenous IAA and ABA didn't affect leaf type of HX3 and rl. CONCLUSIONS: Our results suggest abnormal cytokinin metabolism caused rolled and narrow leaf in rl, and provide valuable clues for further understanding the mechanisms underlying leaf development in soybean.

Genome-wide association study and haplotype analysis reveal novel candidate genes for resistance to powdery mildew in soybean.

Powdery mildew disease (PMD) is caused by the obligate biotrophic fungus Microsphaera diffusa Cooke & Peck (M. diffusa) and results in significant yield losses in soybean (Glycine max (L.) Merr.) crops. By identifying disease-resistant genes and breeding soybean accessions with enhanced resistance, we can effectively mitigate the detrimental impact of PMD on soybeans. We analyzed PMD resistance in a diversity panel of 315 soybean accessions in two locations over 3 years, and candidate genes associated with PMD resistance were identified through genome-wide association studies (GWAS), haplotype analysis, qRT-PCR, and EMS mutant analysis. Based on the GWAS approach, we identified a region on chromosome 16 (Chr16) in which 21 genes form a gene cluster that is highly correlated with PMD resistance. In order to validate and refine these findings, we conducted haplotype analysis of 21 candidate genes and indicated there are single nucleotide polymorphisms (SNPs) and insertion-deletions (InDels) variations of Glyma.16G214000, Glyma.16G214200, Glyma.16G215100 and Glyma.16G215300 within the coding and promoter regions that exhibit a strong association with resistance against PMD. Subsequent structural analysis of candidate genes within this cluster revealed that in 315 accessions, the majority of accessions exhibited resistance to PMD when Glyma.16G214300, Glyma.16G214800 and Glyma.16G215000 were complete; however, they demonstrated susceptibility to PMD when these genes were incomplete. Quantitative real-time PCR assays (qRT-PCR) of possible candidate genes showed that 14 candidate genes (Glyma.16G213700, Glyma.16G213800, Glyma.16G213900, Glyma.16G214000, Glyma.16G214200, Glyma.16G214300, Glyma.16G214500, Glyma.16G214585, Glyma.16G214669, Glyma.16G214700, Glyma.16G214800, Glyma.16G215000, Glyma.16G215100 and Glyma.16G215300) were involved in PMD resistance. Finally, we evaluated the PMD resistance of mutant lines from the Williams 82 EMS mutations library, which revealed that mutants of Glyma.16G214000, Glyma.16G214200, Glyma.16G214300, Glyma.16G214800, Glyma.16G215000, Glyma.16G215100 and Glyma.16G215300, exhibited sensitivity to PMD. Combined with the analysis results of GWAS, haplotypes, qRT-PCR and mutants, the genes Glyma.16G214000, Glyma.16G214200, Glyma.16G214300, Glyma.16G214800, Glyma.16G215000, Glyma.16G215100 and Glyma.16G215300 were identified as highly correlated with PMD resistance. The candidate genes identified above are all NLR family genes, and these discoveries deepen our understanding of the molecular basis of PMD resistance in soybeans and will be useful for guiding breeding strategies.

Untargeted metabolomics revealed the effect of soybean metabolites on poly(γ-glutamic acid) production in fermented natto and its metabolic pathway.

BACKGROUND: Natto mucus is mainly composed of poly(γ-glutamic acid) (γ-PGA), which affects the sensory quality of natto and has some effective functional activities. The soybean metabolites that cause different γ-PGA contents in different fermented natto are unclear. RESULTS: In this study, we use untargeted metabolomics to analyze the metabolites of high-production γ-PGA natto and low-production γ-PGA natto and their fermented substrate soybean. A total of 257 main significantly different metabolites with the same trend among the three comparison groups were screened, of which 114 were downregulated and 143 were upregulated. Through the enrichment of metabolic pathways, the metabolic pathways with significant differences were purine metabolism, nucleotide metabolism, fructose and mannose metabolism, anthocyanin biosynthesis, isoflavonoid biosynthesis and the pentose phosphate pathway. CONCLUSION: For 114 downregulated main significantly different metabolites with the same trend among the three comparison groups, Bacillus subtilis (natto) may directly decompose them to synthesize γ-PGA. Adding downregulated substances before fermentation or cultivating soybean varieties with the goal of high production of such substances has a great effect on the production of γ-PGA by natto fermentation. The enrichment analysis results showed the main pathways affecting the production of γ-PGA by Bacillus subtilis (natto) using soybean metabolites, which provides a theoretical basis for the production of γ-PGA by soybean and promotes the diversification of natto products. © 2023 Society of Chemical Industry.

Effects of Biological Nitrogen Metabolism on Glufosinate-Susceptible and -Resistant Goosegrass (Eleusine indica L.).

Glufosinate is a broad-spectrum herbicide used to control most weeds in agriculture worldwide. Goosegrass (Eleusine indica L.) is one of the top ten malignant weeds across the world, showing high tolerance to glufosinate via different mechanisms that are not yet fully understood. This study revealed that nitrogen metabolism could be a target-resistant site, providing clues to finally clarify the mechanism of glufosinate resistance in resistant goosegrass populations. Compared to susceptible goosegrass (NX), the resistant goosegrass (AUS and CS) regarding the stress of glufosinate showed stronger resistance with lower ammonia contents, higher target enzyme GS (glutamine synthetase) activity, and lower GOGAT (glutamine 2-oxoglutarate aminotransferase) activity. The GDH (glutamate dehydrogenase) activity of another pathway increased, but its gene expression was downregulated in resistant goosegrass (AUS). Analyzing the transcriptome and proteome data of goosegrass under glufosinate stress at 36 h showed that the KEGG pathway of the nitrogen metabolism was enriched in glufosinate-susceptible goosegrass (NX), but not in glufosinate-resistant goosegrass (CS and AUS). Several putative target genes involved in glufosinate stress countermeasures were identified. This study provides specific insights into the nitrogen metabolism of resistant goosegrass, and gives a basis for future functional verification of glufosinate-tolerance genes in plants.

The impact of local government competition and green technology innovation on economic low-carbon transition: new insights from China.

The government-led Chinese economic development system determines that local government competition is a significant factor affecting the economic low-carbon transition. Driving an economic development mode with green technology innovation as the core is the critical path to realizing an economic low-carbon transition. Consequently, it is of significant practical relevance to investigate the impact of local government competition and green technology innovation on the economic low-carbon transition under the government-led Chinese economic development system. This paper systematically explores the nexus between green technology innovation and economic low-carbon transition in terms of local government competition perspective using the system generalized method of moments, panel threshold model, and geographically weighted regression on the basis of a dataset of 30 provincial administrative areas in China from a period of 2006-2019. The results indicate that green technology innovation significantly promotes the economic low-carbon transition. Local government competition not only significantly dampens the economic low-carbon transition but also considerably inhibits the positive effect of green technology innovation on the economic low-carbon transition. A significant N-shaped association is evident between green technology innovation and the economic low-carbon transition when green technology innovation is applied as a threshold, while such association is insignificant when local government competition is used as a threshold. Compared with high-competition intensity areas, green technology innovation promotes economic low-carbon transition weaker in low- competition intensity areas, while local government competition inhibits economic low-carbon transition stronger. However, local government competition significantly inhibits the positive effect of green technology innovation on the economic low-carbon transition in low-competition intensity areas, while insignificant in high-competition intensity areas. The geographically weighted regression technique as a whole also verified the above results. Therefore, policymakers should not only increase research and development investment in green technologies, but also develop a regionally linked low-carbon emission reduction system to avoid ineffective competition among governments to facilitate the earlier fulfillment of the "dual carbon" goal.

Green Total Factor Productivity Growth: Policy-Guided or Market-Driven?

The green growth mode of modern economy is affected by both policy and market, but previous studies have lacked a comparison between the two effects on green economy development. Which is the leading factor of green growth: policy or market? Using the Panel Smooth Transition Regression (PSTR) model and the twelve-year data of more than 200 prefecture-level cities in China, we compared and analyzed the linear and non-linear effects of environmental regulation and marketization degree on green total factor productivity (GTFP). The results show that: (1) both environmental regulation and marketization degree have a non-linear promoting effect on GTFP. (2) GTFP is mainly market-driven rather than policy-guided. (3) Environmental regulation and marketization promote the improvement of GTFP through the industrial upgrading effect and the innovation development effect, respectively. This paper makes up for the comparative analysis gap of factors in the field of green growth and extends from the single determination of influencing factors to the importance of the comparison of influencing factors with the transition perspective. The conclusions provide a reference for the green development of countries and regions, emphasizing the importance of green development policies adapting to local conditions and time and providing evidence for market-oriented green economy development.

Functional Characterization of the Lysine-Specific Histone Demethylases Family in Soybean.

Histone modifications, such as methylation and demethylation, have crucial roles in regulating chromatin structure and gene expression. Lysine-specific histone demethylases (LSDs) belong to the amine oxidase family, which is an important family of histone lysine demethylases (KDMs), and functions in maintaining homeostasis of histone methylation. Here, we identified six LSD-like (LDL) genes from the important leguminous soybean. Phylogenetic analyses divided the six GmLDLs into four clusters with two highly conserved SWRIM and amine oxidase domains. Indeed, demethylase activity assay using recombinant GmLDL proteins in vitro demonstrated that GmLDLs have demethylase activity toward mono- and dimethylated Lys4 but not trimethylated histone 3, similar to their orthologs previously reported in animals. Using real-time PCR experiments in combination with public transcriptome data, we found that these six GmLDL genes exhibit comparable expressions in multiple tissues or in response to different abiotic stresses. Moreover, our genetic variation investigation of GmLDL genes among 761 resequenced soybean accessions indicates that GmLDLs are well conserved during soybean domestication and improvement. Taken together, these findings demonstrate that GmFLD, GmLDL1a, and GmLDL1b are bona fide H3K4 demethylases towards H4K4me1/2 and GmLDLs exist in various members with likely conserved and divergent roles in soybeans.

Interaction between Digital Economy and Environmental Pollution: New Evidence from a Spatial Perspective.

The digital economy and the green economy are two major issues for economic recovery in the post epidemic era. From spatial interaction spillover, we analyze and measure the relationships between the digital economy and environmental pollution in 287 prefecture-level cities in China from 2008 to 2018 using simultaneous spatial equations and the generalized 3-stage least square (GS3SLS) method. The results show that: (1) there is a reverse and complex spatio-temporal evolution of the digital economy and environmental pollution in Chinese cities. (2) There is a spatial interaction spillover effect between the digital economy and environmental pollution. Local digital economy and environmental pollution inhibit each other. The digital economy and environmental pollution have a significant spatial spillover. The digital economy of surrounding regions has a suppressive effect on local environmental pollution. The environmental pollution of surrounding cities has a crowding-out effect on the local digital economy. (3) Digital economy suppresses environmental pollution through the green development effect and innovative development effect; environmental pollution suppresses the digital economy through the talent crowding out effect and the policy tightening effect. The conclusion of this paper provides evidence for the coupling and coordinated development between the digital and green economy, which is of great significance for promoting the transformation of economic development modes and realizing green and high-quality development.

Quantitative Trait Locus Mapping of Seed Vigor in Soybean under -20 °C Storage and Accelerated Aging Conditions via RAD Sequencing.

Due to its fast deterioration, soybean (Glycine max L.) has an inherently poor seed vigor. Vigor loss occurring during storage is one of the main obstacles to soybean production in the tropics. To analyze the genetic background of seed vigor, soybean seeds of a recombinant inbred line (RIL) population derived from the cross between Zhonghuang24 (ZH24, low vigor cultivar) and Huaxia3hao (HX3, vigorous cultivar) were utilized to identify the quantitative trait loci (QTLs) underlying the seed vigor under -20 °C conservation and accelerated aging conditions. According to the linkage analysis, multiple seed vigor-related QTLs were identified under both -20 °C and accelerated aging storage. Two major QTLs and eight QTL hotspots localized on chromosomes 3, 6, 9, 11, 15, 16, 17, and 19 were detected that were associated with seed vigor across two storage conditions. The indicators of seed vigor did not correlate well between the two aging treatments, and no common QTLs were detected in RIL populations stored in two conditions. These results indicated that deterioration under accelerated aging conditions was not reflective of natural aging at -20 °C. Additionally, we suggest 15 promising candidate genes that could possibly determine the seed vigor in soybeans, which would help explore the mechanisms responsible for maintaining high seed vigor.

Spatial Interaction Spillover Effects between Digital Financial Technology and Urban Ecological Efficiency in China: An Empirical Study Based on Spatial Simultaneous Equations.

As a core component of the digital economy, digital financial technology has a complex interactive and interdependent relationship with ecological efficiency. From the holistic spatial interaction perspective, this paper uses spatial simultaneous equations and generalized spatial three-stage least squares (GS3SLS) to analyze the spatial interaction spillovers between digital financial technology and urban ecological efficiency based on data from 284 Cities in China from 2008 to 2018. The results show that: (1) Digital financial technology and urban ecological efficiency promote each other, and the latter is relatively dominant. (2) Both digital financial technology and urban ecological efficiency have significant spatial spillover effects. (3) Digital financial technology in surrounding cities has a restraining effect on local ecological efficiency, and the improvement of ecological efficiency in surrounding cities has a siphon effect on local digital financial technology. (4) There is spatial and period heterogeneity in the intensity of the spatial interaction spillover effect between the two. With resources and environment increasingly becoming rigid constraints on economic growth, these findings help identify new drivers of regional ecological efficiency and promote the coordinated development of digital finance and green ecology.

Modulation of evening complex activity enables north-to-south adaptation of soybean.

Soybean, a typical short-day crop, is sensitive to photoperiod, which is a major limiting factor defining its north-to-south cultivation range. The long-juvenile (LJ) trait is controlled primarily by the J locus which has been used for decades by soybean breeders to delay flowering and improve grain yield in tropical regions. The J gene encodes an ortholog of the Arabidopsis Evening Complex (EC) component EARLY FLOWERING 3 (ELF3). To identify modifiers of J, we conducted a forward genetic screen and isolated a mutant (eoj57) that in combination with j has longer flowering delay compared with j single mutant plants. Map-based cloning and genome re-sequencing identified eoj57 (designated as GmLUX2) as an ortholog of the Arabidopsis EC component LUX ARRHYTHMO (LUX). To validate that GmLUX2 is a modifier of J, we used trans-complementation and identified a natural variant allele with a similar phenotype. We also show that GmLUX2 physically interacts with GmELF3a/b and binds DNA, whereas the mutant and natural variant are attenuated in both activities. Transcriptome analysis shows that the GmLUX2-GmELF3a complex co-regulates the expression of several circadian clock-associated genes and directly represses E1 expression. These results provide mechanistic insight into how GmLUX2-GmELF3 controls flowering time via synergistic regulation of gene expression. These novel insights expand our understanding of the regulation of the EC complex, and facilitate the development of soybean varieties adapted for growth at lower latitudes.

QTL mapping for aluminum tolerance in RIL population of soybean (Glycine max L.) by RAD sequencing.

Aluminum (Al3+) toxicity is a typical abiotic stress that severely limits crop production in acidic soils. In this study, an RIL (recombinant inbred line, F12) population derived from the cross of Zhonghuang 24 (ZH 24) and Huaxia 3 (HX 3) (160 lines) was tested using hydroponic cultivation. Relative root elongation (RRE) and apical Al3+ content (AAC) were evaluated for each line, and a significant negative correlation was detected between the two indicators. Based on a high-density genetic linkage map, the phenotypic data were used to identify quantitative trait loci (QTLs) associated with these traits. With composite interval mapping (CIM) of the linkage map, five QTLs that explained 39.65% of RRE and AAC variation were detected on chromosomes (Chrs) Gm04, Gm16, Gm17 and Gm19. Two new QTLs, qRRE_04 and qAAC_04, were located on the same region of bin93-bin94 on Chr Gm04, which explained 7.09% and 8.98% phenotypic variation, respectively. Furthermore, the results of the expression analysis of candidate genes in the five genetic regions of the QTLs showed that six genes (Glyma.04g218700, Glyma.04g212800, Glyma.04g213300, Glyma.04g217400, Glyma.04g216100 and Glyma.04g220600) exhibited significant differential expression between the Al3+ treatment and the control of two parents. The results of qRT-PCR analysis indicated that Glyma.04g218700 was upregulated by Al3+ treatment with the hundreds-fold increased expression level and may be a candidate gene with potential roles in the response to aluminum stress. Therefore, our efforts will enable future functional analysis of candidate genes and will contribute to the strategies for improvement of aluminum tolerance in soybean.

The differences of cell wall in roots between two contrasting soybean cultivars exposed to cadmium at young seedlings.

The plant root cell wall (CW) is the first structure that comes into contact with extracellular cadmium (Cd), and it plays an important role in the absorption, immobilization, and translocation of Cd in the roots. However, the differences in the cell wall components between Cd-tolerant and Cd-sensitive cultivars are unclear. A hydroponic experiment was carried out to investigate the differences in the concentrations of Cd, total sugars, and uronic acid in pectin, hemicellulose 1, hemicellulose 2, cellulose, and lignin, as well as pectin methylesterase enzyme activity (PME) in the roots of two soybean cultivars that differ with respect to Cd tolerance exposed to 0 and 23 µM Cd treatments. The bound forms of Cd in the roots were found to differ between the two soybean genotypes; 50.2% of the Cd in the root cell wall accumulates in the pectin in the highly Cd-tolerant and low Cd-accumulating cultivar HX3, while 50.6% of the root cell wall Cd accumulates in cellulose in the Cd-sensitive and high Cd-accumulating cultivar BX10. The total sugar and uronic acid concentrations of the cell wall components increased in response to Cd stress, while the concentrations of total sugars and uronic acid in BX10 were higher than in HX3 (except for hemicellulose 1). Increased demethylation of pectin may be the main reason that Cd is mainly concentrated in the primary wall in HX3, because the PME activity was higher in HX3 than it was in BX10 under Cd treatment. Furthermore, BX10 had a higher lignin concentration after Cd treatment, and showed the same change in cellulose. Cd in the root cell wall of BX10 was fixed in the secondary cell wall, which may be a result of the coupling to cellulose and lignin. In conclusion, root cell walls in soybean cultivars that differ in Cd tolerance may possess different mechanisms to prevent Cd from entering cells, and the sequestration of Cd in different cell wall components may determine the differences in Cd tolerance between the two genotypes.

Genome-wide characterization of soybean P 1B -ATPases gene family provides functional implications in cadmium responses.

BACKGROUND: The P1B-ATPase subfamily is an important group involved in transporting heavy metals and has been extensively studied in model plants, such as Arabidopsis thaliana and Oryza sativa. Emerging evidence indicates that one homolog in Glycine max is also involved in cadmium (Cd) stress, but the gene family has not been fully investigated in soybean. RESULTS: Here, we identified 20 heavy metal ATPase (HMA) family members in the soybean genome, presented as 10 paralogous pairs, which is significantly greater than the number in Arabidopsis or rice, and was likely caused by the latest whole genome duplication event in soybean. A phylogenetic analysis divided the 20 members into six groups, each having conserved or divergent gene structures and protein motif patterns. The integration of RNA-sequencing and qRT-PCR data from multiple tissues provided an overall expression pattern for the HMA family in soybean. Further comparisons of expression patterns and the single nucleotide polymorphism distribution between paralogous pairs suggested functional conservation and the divergence of HMA genes during soybean evolution. Finally, analyses of the HMAs expressed in response to Cd stress provided evidence on how plants manage Cd tolerance, at least in the two contrasting soybean genotypes examined. CONCLUSIONS: The genome-wide identification, chromosomal distribution, gene structures, and evolutionary and expression analyses of the 20 HMA genes in soybean provide an overall insight into their potential involvement in Cd responses. These results will facilitate further research on the HMA gene family, and their conserved and divergent biological functions in soybean.

Root morphological responses of five soybean [Glycine max (L.) Merr] cultivars to cadmium stress at young seedlings.

To examine the differences in root morphological responses of soybean cultivars with different cadmium (Cd) tolerance and accumulation to Cd stress, the biomass, Cd concentration, and root morphological features of five soybean cultivars were determined under 0, 9, 23, 45, and 90 µM Cd stress via hydroponic experiments. Significantly genotypic differences in Cd tolerance and Cd concentration were observed between five soybean cultivars at four Cd levels. For Cd tolerance, HX3 showed a strong Cd tolerance with tolerance indexes of shoot biomass at 92.49, 76.44, 60.21, and 46.45% after 18 days at four Cd levels, and others had similar weak tolerance at young seedling. For Cd accumulation, Cd concentration in roots showed far higher than that in shoots. The different accumulation features in roots and shoots among five cultivars were found at four Cd levels. Comparing with the control, the total root length (RL), root surface area (SA), and root volume (RV) of all cultivars were decreased significantly at four Cd levels. Tolerant cultivar HX3 had the largest root system and sensitive cultivar BX10 had the smallest root system at young seedling stage. Correlation analysis indicated that RL, SA, and RV were positively correlated with root biomass and shoot biomass under 9 and 23 µM Cd treatments, but root average diameter (RD) was negatively correlated with shoot biomass and root biomass only under 9 µM Cd treatments, while RL and SA were negatively correlated with root Cd concentration under 23 and 45 µM Cd treatments. The results suggested that root morphological traits were closely related to Cd tolerance at young seedlings under Cd treatments.

Comparison of subcellular distribution and chemical forms of cadmium among four soybean cultivars at young seedlings.

The hydroponic experiment was carried out to investigate the Cd subcellular distribution and chemical forms in roots and shoots among four soybean seedling cultivars with two Cd treatments. HX3 and GC8, two tolerant and low-grain-Cd-accumulating cultivars, had the lowest Cd concentration in roots and high Cd concentration in shoots, while BX10 and ZH24, two sensitive and high-grain-Cd-accumulating cultivars, had the highest Cd concentration in roots and the lowest Cd concentration in shoots at young seedling stage. Furthermore, the sequence of Cd subcellular distribution in roots at two Cd levels was cell wall (53.4-75.5 %) > soluble fraction (15.8-40.4 %) > organelle fraction (2.0-14.7 %), but in shoots, was soluble fraction (39.3-74.8 %) > cell wall (16.0-52.0 %) > organelle (4.8-19.5 %). BX10 and ZH24 had higher Cd concentration in all subcellular fractions in roots, but HX3 and GC8 had higher Cd concentration of soluble fraction in shoots. The sequence of Cd chemical forms in roots was FNacl (64.1-79.5 %) > FHAC (3.4-21.5 %) > Fd-H2O (3.6-13.0 %) > Fethanol (1.4-21.8) > FHCl (0.3-1.6 %) > Fother (0.2-1.4 %) at two Cd levels but, in shoots, was FNacl (19.7-51.4 %) ≥ FHAC (10.2-31.4 %) ≥ Fd-H2O (8.8-28.2 %) ≥ Fethanol (8.9-38.6 %) > FHCl (0.2-9.6 %) > Fother (2.5-11.2 %). BX10 and ZH24 had higher Cd concentrations in each extracted solutions from roots, but from shoots for GC8 and HX3. Taken together, the results uncover that root cell walls and leaf vacuoles might play important roles in Cd detoxification and limiting the symplastic movement of Cd.

Transcriptome profiling to discover putative genes associated with paraquat resistance in goosegrass (Eleusine indica L.).

BACKGROUND: Goosegrass (Eleusine indica L.), a serious annual weed in the world, has evolved resistance to several herbicides including paraquat, a non-selective herbicide. The mechanism of paraquat resistance in weeds is only partially understood. To further study the molecular mechanism underlying paraquat resistance in goosegrass, we performed transcriptome analysis of susceptible and resistant biotypes of goosegrass with or without paraquat treatment. RESULTS: The RNA-seq libraries generated 194,716,560 valid reads with an average length of 91.29 bp. De novo assembly analysis produced 158,461 transcripts with an average length of 1153.74 bp and 100,742 unigenes with an average length of 712.79 bp. Among these, 25,926 unigenes were assigned to 65 GO terms that contained three main categories. A total of 13,809 unigenes with 1,208 enzyme commission numbers were assigned to 314 predicted KEGG metabolic pathways, and 12,719 unigenes were categorized into 25 KOG classifications. Furthermore, our results revealed that 53 genes related to reactive oxygen species scavenging, 10 genes related to polyamines and 18 genes related to transport were differentially expressed in paraquat treatment experiments. The genes related to polyamines and transport are likely potential candidate genes that could be further investigated to confirm their roles in paraquat resistance of goosegrass. CONCLUSION: This is the first large-scale transcriptome sequencing of E. indica using the Illumina platform. Potential genes involved in paraquat resistance were identified from the assembled sequences. The transcriptome data may serve as a reference for further analysis of gene expression and functional genomics studies, and will facilitate the study of paraquat resistance at the molecular level in goosegrass.

Metal pollution (Cd, Pb, Zn, and As) in agricultural soils and soybean, Glycine max, in southern China.

Crops produced on metal-polluted agricultural soils may lead to chronic toxicity to humans via the food chain. To assess metal pollution in agricultural soils and soybean in southern China, 30 soybean grain samples and 17 soybean-field soil samples were collected from 17 sites in southern China, and metal concentrations of samples were analyzed by graphite furnace atomic absorption spectrophotometer. The integrated pollution index was used to evaluate if the samples were contaminated by Cd, Pb, Zn and As. Results showed that Cd concentration of 12 samples, Pb concentration of 2 samples, Zn concentration of 2 samples, and As concentrations of 2 samples were above the maximum permissible levels in soils. The integrated pollution index indicated that 11 of 17 soil samples were polluted by metals. Metal concentrations in soybean grain samples ranged from 0.11 to 0.91 mg kg(-1) for Cd; 0.34 to 2.83 mg kg(-1) for Pb; 42 to 88 mg kg(-1) for Zn; and 0.26 to 5.07 mg kg(-1) for As, which means all 30 soybean grain samples were polluted by Pb, Pb/Cd, Cd/Pb/As or Pb/As. Taken together, our study provides evidence that metal pollution is an important concern in agricultural soils and soybeans in southern China.

Identification and comparative analysis of cadmium tolerance-associated miRNAs and their targets in two soybean genotypes.

MicroRNAs (miRNAs) play crucial roles in regulating the expression of various stress responses genes in plants. To investigate soybean (Glycine max) miRNAs involved in the response to cadmium (Cd), microarrays containing 953 unique miRNA probes were employed to identify differences in the expression patterns of the miRNAs between different genotypes, Huaxia3 (HX3, Cd-tolerant) and Zhonghuang24 (ZH24, Cd-sensitive). Twenty six Cd-responsive miRNAs were identified in total. Among them, nine were detected in both cultivars, while five were expressed only in HX3 and 12 were only in ZH24. The expression of 16 miRNAs was tested by qRT-PCR and most of the identified miRNAs were found to have similar expression patterns with microarray. Three hundred and seventy six target genes were identified for 204 miRNAs from a mixture degradome library, which was constructed from the root of HX3 and ZH24 with or without Cd treatment. Fifty five genes were identified to be cleaved by 14 Cd-responsive miRNAs. Gene ontology (GO) annotations showed that these target transcripts are implicated in a broad range of biological processes. In addition, the expression patterns of ten target genes were validated by qRT-PCR. The characterization of the miRNAs and the associated target genes in response to Cd exposure provides a framework for understanding the molecular mechanism of heavy metal tolerance in plants.

OsDREB2A, a rice transcription factor, significantly affects salt tolerance in transgenic soybean.

The dehydration responsive element binding (DREB) transcription factors play an important role in regulating stress-related genes. OsDREB2A, a member of the DREBP subfamily of AP2/ERF transcription factors in rice (Oryza sativa), is involved in the abiotic stress response. OsDREB2A expression is induced by drought, low-temperature and salt stresses. Here, we report the ability of OsDREB2A to regulate high-salt response in transgenic soybean. Overexpressing OsDREB2A in soybeans enhanced salt tolerance by accumulating osmolytes, such as soluble sugars and free proline, and improving the expression levels of some stress-responsive transcription factors and key genes. The phenotypic characterization of transgenic soybean were significantly better than those of wild-type (WT). Electrophoresis mobility shift assay (EMSA) revealed that the OsDREB2A can bind to the DRE core element in vitro. These results indicate that OsDREB2A may participate in abiotic stress by directly binding with DRE element to regulate the expression of downstream genes. Overexpression of OsDREB2A in soybean might be used to improve tolerance to salt stress.