Improving the production of broiler chicken meat has been a goal of broiler breeding programs worldwide for many years.  However, the genetic architectures of skeletal muscle production traits in chickens have not yet been fully elucidated.  In the present study, a total of 519 F₂ birds, derived from a cross of Arbor Acres broiler and Baier layer, were re-sequenced (26 F₀ individuals were re-sequenced at a 10-fold depth; 519 F₂ individuals were re-sequenced at a 3-fold depth) and the coupling of genome-wide association study (GWAS) and selection signatures (F_ST (fixation index) and θ_π (nucleotide diversity)) was carried out to pinpoint the associated loci and genes that contribute to pectoral muscle weight (PMW) and thigh muscle weight (TMW).  A total of 7 890 258 single nucleotide polymorphisms (SNPs) remained to be analyzed after quality control and imputation.  The integration of GWAS and selection signature analyses revealed that genetic determinants responsible for skeletal muscle production traits were mainly localized on chromosomes 1 (168.95–172.43 Mb) and 4 (74.37–75.23 Mb).  A total of 17 positional candidate genes (PCGs) (LRCH1, CDADC1, CAB39L, LOC112531568, LOC112531569, FAM124A, FOXO1, NBEA, GPALPP1, RUBCNL, ARL11, KPNA3, LHFP, GBA3, LOC112532426, KCNIP4, and SLIT2) were identified in these regions.  In particular, KPNA3 and FOXO1 were the most promising candidates for meat production in chickens.  These findings will help enhance our understanding of the genetic architecture of chicken muscle production traits, and the significant SNPs identified could be promising candidates for integration into practical breeding programs such as genome-wide selection (GS) to improve the meat yield of chickens.

    The danger of mycotoxin contamination entering the food supply through post-harvest infection is of perennial concern to food safety experts. To explore the distribution of Penicillium expansum and diffusion of its mycotoxin, patulin, in blue mold-damaged pears, Pyrus bretschneideri Rehd. cv. Yali obtained from markets and orchards in China were artificially inoculated with P. expansum and assayed for patulin accumulation and degree of fungal colonization. The inoculated pears were incubated until the lesions were 5, 10, 20, or 30 mm in diameter. We sampled tissue at a range of distances from the lesion, measured the spread of Penicillium by plate colony-counting methods, and used UHPLC-MS/MS to detect and quantify the patulin concentration. More P. expansum colony-forming units were isolated from pears with a higher degree of decay. Farther from the lesion, the fewer P. expansum colonies were observed, and the lower the patulin content detected. We found a significant difference in the patulin content between samples due to lesion size, and also in tissue sampled 10 mm away from the lesion. In consideration of this finding, to ensure food safety, we recommend that when a blue mold rot lesion on pear is 5, 10, or 20 mm in diameter, 20, 30, and 40 mm beyond the lesion should be removed, respectively. If a lesion surpasses 30 mm in diameter, the whole pear should be thrown away.

Auxin has been identified to play critical roles in regulating plant growth and development.  The polar transport of auxin is regulated by auxin transporters.  In the present study, an auxin efflux carrier gene MdPIN1 was cloned from Malus×domestic, Royal Gala, and introduced into wild-type Arabidopsis thaliana (Col-0).  The transgenic plants exhibited the phenotype of inhibition of primary root (PR) elongation and increased lateral root (LR) number in compared with Col-0.  Overexpression of MdPIN1 affected auxin transport, and enhanced phototropic responses and geotropism reaction, whereas had no significant difference in the auxin biosynthesis.  These findings suggest that the MdPIN1 gene plays a vital role in auxin transport and root development.

Litter uniformity, which is usually represented by within-litter weight coefficient of variation at birth (CVB), could influence litter performance of sows and the profitability of pig enterprises. The objective of this study was to characterize CVB and its effect on other reproductive traits in Large White sows. Genetic parameters and genetic correlation of the reproductive traits, including CVB, within-litter weight coefficient of variation at three weeks (CVT), total number born (TNB), number born alive (NBA), number born dead (NBD), gestation length (GL), piglet mortality at birth (M₀), piglet mortality at three weeks (M₃), total litter weight at birth (TLW0), and total litter weight at three weeks (TLW₃) were estimated for 2 032 Large White litters. The effects of parity and classified litter size on CVB, CVT, TNB, NBA, NBD, GL, M₀, M₃, TLW₀, and TLW₃ were also estimated. The heritabilities of these reproductive traits ranged from 0.06 to 0.17, with the lowest heritability for CVB and the highest heritability for TLW0. Phenotypic and genetic correlations between these reproductive traits were low to highly positive and negative (ranging from −0.03 to 0.93, and −0.53 to 0.93, respectively). The genetic correlations between TNB and CVB, and between M₀ and CVB were 0.32 and 0.29, respectively. In addition, CVB was significantly influenced by parity and litter size class (P<0.05). All the results suggest that piglet uniformity should be maintained in pig production practices and pig breeding programs.

    It is a novel idea to make steamed bread by adding potato flour into wheat flour considering the production and nutritional factors of potato. In this study, the influence of potato flour (0–35%) on dough rheology and quality of steamed bread were investigated. Potato flour addition significantly influenced the dough rheological properties and steamed bread quality, such as increased water absorption, the maximum gaseous release height, total volume of CO₂ and hardness, while decreased dough stability and specific volume of steamed bread. Moreover, correlation analysis suggested that dough height at the maximum development time, dough stability, water absorption and the phase tangent can be used for predicting the technological quality of steamed bread. Potato-wheat steamed bread had higher dietary fibre, ash content and antioxidant activity than those of wheat steamed bread. The estimated glycemic index decreased from 73.63 (0%) to 60.01 (35%). Considering the sensory evaluation, the steamed bread with 20% potato flour is acceptable. In conclusion, adding appropriate quantity of potato flour to wheat flour for steamed bread production will not only maintain the technological quality, but also can improve the nutritional value of the steamed bread.

   Ear size exhibits remarkable diversity in pig breeds. LEM domain-containing 3 (LEMD3) on chromosome 5 is considered as an important candidate for porcine ear size. This is the first study on cloning and characterization of LEMD3 cDNA. The complete cDNA contains 4 843 bp, including a 2 736-bp open reading frame (ORF), a 37-bp 5´-untranslated region (UTR) and a 2 070-bp 3´-UTR. The complete LEMD3 gene is 126 241-bp and contains 13 exons and 12 introns. The ORF encodes a deduced LEMD3 protein of 911 amino acids, which shares 82–94% nucleic acid and 51–96% amino acid identity with other species. A phylogenetic tree constructed based on the amino acid sequences revealed that the porcine LEMD3 protein was closely related with cattle LEMD3. Resequencing of the ORF and promoter of LEMD3 from Minzhu pig and Large White revealed three single nucleotide polymorphisms (SNPs): L964C>A in the complete coding region, L4625A>G in the 3´ UTR, and L-394T>C in the promoter region. Genome-wide association study (GWAS) revealed that all of SNPs were shown significant association with ear size in Large White×Minzhu pig intercross population. With conditional GWAS, –log₁₀(P-value) decreased by more than 80% when each of three SNPs was included as a fixed effect. These results suggested direct involvement of LEMD3 or close linkage to the causative mutation for ear size. The findings of this study might form the basis for understanding the genetic mechanism of ear size variation in pigs and provide potential molecular markers for screening ear size diversity in pig breeds.

The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV) and porcine circovirus type 2 (PCV-2) in pigs in China. A total of 372 tissues or serum samples collected from pigs distributed in 9 provinces/ municipalities of China during the period from February 2011 to November 2012 were assayed for T. gondii antigens and antibodies using enzyme linked immunosorbent assay (ELISA) technique, while the PCR was designed for the detection of the PRRSV, CSFV and PCV-2, respectively. The total positive rate of T. gondii, PRSSV, CSFV and PCV-2 was 9.14% (34/372), 50.00% (186/372), 37.10% (138/372) and 3.23% (12/372), respectively. Among the 34 T. gondii positive samples, 26 samples were simultaneously infected with T. gondii and viruses, while the remaining eight samples were infected with T. gondii alone. In addition, the co-infection rate of T. gondii with PRSSV, T. gondii with PRSSV and CSFV, T. gondii with PRSSV and PCV-2, T. gondii with CSFV and PCV-2, T. gondii with PRSSV, CSFV and PCV-2 was 1.61% (6/372), 4.03% (15/372), 0.27% (1/372), 0.27% (1/372) and 0.81% (3/372), respectively. The results of the present survey revealed that PRRSV and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China, and both of them could increase the chances of T. gondii infection in pig. This is the first report of T. gondii co-infections with viruses in pigs. It is very important to understand the interactions of parasite and virus, and can be used as reference data for the control and prevention of co-infections of T. gondii and viruses in pigs.

The construction of high density genetic linkage map provides a powerful tool to detect and map quantitative trait loci (QTLs) controlling agronomically important traits. In this study, simple sequence repeat (SSR) markers and Illumina 9K iSelect single nucleotide polymorphism (SNP) genechip were employed to construct one genetic linkage map of common wheat (Triticum aestivum L.) using 191 recombinant inbred lines (RILs) derived from cross Yu 8679×Jing 411. This map included 1 901 SNP loci and 178 SSR loci, covering 1 659.9 cM and 1 000 marker bins, with an average interval distance of 1.66 cM. A, B and D genomes covered 719.1, 703.5 and 237.3 cM, with an average interval distance of 1.66, 1.45 and 2.9 cM, respectively. Notably, the genetic linkage map covered 20 chromosomes, with the exception of chromosome 5D. Bioinformatics analysis revealed that 1 754 (92.27%) of 1 901 mapped SNP loci could be aligned to 1 215 distinct wheat unigenes, among which 1 184 (97.4%) were located on one single chromosome, and the rest 31 (2.6%) were located on 2 to 3 chromosomes. By performing in silico comparison, 214 chromosome deletion bin-mapped expressed sequence tags (ESTs), 1 043 Brachypodium genes and 1 033 rice genes were further added onto the genetic linkage map. This map not only integrated genetic and physical maps, SSR and SNP loci, respectively, but also provided the information of Brachypodium and rice genes corresponding to 1 754 SNP loci. Therefore, it will be a useful tool for comparative genomics analysis, fine mapping of QTL/gene controlling agronomically important traits and marker-assisted selection breeding in wheat.

Water and nitrogen are primary limiting factors in semiarid grassland ecosystems. Our knowledge is still poor regarding the interactive effects of water and N addition on soil microbial communities, although this information is crucial to reveal the mechanisms of the terrestrial ecosystem response to global changes. We addressed this problem by conducting a field experiment with a 15% surplus of the average rainfall under three levels of N addition (50, 100, and 200 kg N ha–1 yr–1) in two consecutive years in Inner Mongolia, China. Microbial community composition and functional diversity were analyzed based on phospholipid fatty acids (PLFA) and BIOLOG techniques, respectively. The results showed that water addition did not affect the soil microbial community composition, but much more yearly precipitation generally decreased the PLFA concentration, which implied a fast response of soil microbes to changes of water condition. Soil fungi was depressed only by N addition at the high level (200 kg N ha–1 yr–1) and without hydrologic leaching, while Gram-negative bacteria was suppressed probably by plant competition at high level N addition but with hydrologic leaching. The study found unilateral positive/negative interactions between water and N addition in affecting soil microbial community, however, climate condition (precipitation) could be a significant factor in disturbing the interactions. This study highlighted that: (1) The sustained effect of pulsed water addition was minimal on the soil microbial community composition but significant on the microbial community functional diversity and (2) the complex interaction between water and N addition on soil microbial community related to the inter-annual variation of the climate and plant response.

This paper reviewed management of pesticide residues in China including laws and regulations, the supervision system, the standard system, and the quality and safety of agricultural products. The process of establishment and internationalization of standards for pesticide residues were also discussed. Results indicate that the progress of the management of pesticide residues has been steadily made in China. However, the following aspects which refer to updates to regulations, supervising efficiency, standard system, risk assessment, international cooperation and communication, should be further improved. China should draw lessons from international experience, and then establish its own management system, which focuses on pesticides controls by strictly following relevant laws and technical standards to ensure the quality and safety of agricultural products.

Although quantitative trait loci (QTLs) for number of thoracic-lumbar vertebrae have been identified on Sus scrofa chromosomes (SSCs) 1 and 7, the influence of these QTLs on the thoracic and lumbar vertebrae is not clear. The aim of this study was to identify single nucleotide polymorphisms (SNPs) associated with total number of thoracic-lumbar vertebrae and for each trait (number of thoracic and lumbar vertebrae) separately. A total of 581 individuals from an F2 Large White×Minzhu population were genotyped using an SNP60K chip. Performing a genome-wide association study (GWAS) for total number of thoracic-lumbar vertebrae, 38 significant SNPs were identified in two QTL regions located on SSC1 and SSC7. Performing a GWAS for number of thoracic vertebrae only, 72 significant SNPs were located on SSC7. While performing a GWAS for number of lumbar vertebrae only, 17 significant SNPs were identified on SSC1. Gene mining suggested that the gene encoding orphan nuclear receptor, germ cell nuclear factor (NR6A1) on SSC1 was a strong candidate affecting the number of lumbar vertebrae in pigs. Additionally, genes encoding vertnin (VRTN), prospero homeobox 2 (PROX2), Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (FOS), and transforming growth factor beta 3 (TGFB3) may be important candidates affecting the number of thoracic vertebrae in pigs. QTLs on SSC1 and SSC7 independently influenced the numbers of thoracic and lumbar vertebrae. These results shed light on the complex genetic background of vertebrae development in pigs.

The aim of this work was to assess the effect of applying three different doses of fluxapyroxad on microbial activity, community structure and functional diversity as measured by respiration, microbial biomass C, phospholipid fatty acid (PLFA) and community-level physiological profiles (CLPPs). Our results demonstrated that substrate-induced respiration (on day 15) and microbial biomass C (on days 7 and 15) were inhibited by fluxapyroxad, but stimulation was observed thereafter. In contrast, fluxapyroxad addition increased the basal respiration and metabolic quotients (qCO2) and respiratory quotients (QR). Analysis of the PLFA profiles revealed that the total and bacterial biomass (both Gram-positive bacteria (GP) and Gram-negative bacteria (GN)) were decreased within the initial 15 days, whereas those as well as the GN/GP ratio were increased at days 30 and 60. Fluxapyroxad input decreased the fungi biomass but increased the bacteria/fungi ratio at all incubation time. Moreover, high fluxapyroxad input (75 mg fluxapyroxad kg–1 soil dry weight) increased the microbial stress level. A principal component analysis (PCA) of the PLFAs revealed that fluxapyroxad treatment significantly shifted the microbial community structure, but all of the observed effects were transient. Biolog results showed that average well color development (AWCD) and functional diversity index (H´) were increased only on day 60. In addition, the dissipation of fluxapyroxad was slow in soil, and the degradation half-lives varied from 158 to 385 days depending on the concentration tested.

Porcine carcass traits and organ weights have important economic roles in the swine industry. A total of 576 animals from a Large White×Minzhu intercross population were genotyped using the Illumina PorcineSNP60K Beadchip and were phenotyped for 10 traits, specifically, backfat thickness (6-7 libs), carcass length, carcass weight, foot weight, head weight, heart weight, leaf fat weight, liver weight, lung weight and slaughter body weight. The genome-wide association study (GWAS) was assessed by Genome Wide Rapid Association using the mixed model and regression-genomic control approach. A total of 31 single nucleotide polymorphisms (SNPs) (with the most significant SNP being MARC0033464, P value=6.80×10-13) were located in a 9.76-Mb (31.24-41.00 Mb) region on SSC7 and were found to be significantly associated with one or more carcass traits and organ weights. High percentage of phenotypic variance explanation was observed for each trait ranging from 31.21 to 67.42%. Linkage analysis revealed one haplotype block of 495 kb, in which the most significant SNP being MARC0033464 was contained, on SSC7 at complete linkage disequilibrium. Annotation of the pig reference genome suggested 6 genes (GRM4, HMGA1, NUDT3, RPS10, SPDEF and PACSIN1) in this candidate linkage disequilibrium (LD) interval. Functional analysis indicated that the HMGA1 gene presents the prime biological candidate for carcass traits and organ weights in pig, with potential application in breeding programs.

In this study, the hydrolysis of the insecticide ethiprole in buffered solutions at pH 4.0, 7.0 and 9.0, respectively, and the degradation and adsorption-desorption behaviors of ethiprole in five agricultural soil samples from China were investigated. The half-lives under anaerobic conditions were faster than that in the aerobic experiment. Ethiprole was relatively stable under both acidic and neutral conditions while it was readily hydrolyzed under alkaling condition. The sorption of ethiprole on five soils was well described by the linear and Freundlich equation and mainly governed by soil organic matter. The exothermic process of ethiprole adsorption can also be well explained by physical adsorption. A weak adsorption capacity was observed in all soils, which could readily lead to leaching problems.

In 2010, Chinese maize yields increased from 961.5 kg ha-1 in 1949 to 5 453.8 kg ha-1. This increase is the result of genetic improvements, an increase in nitrogen application, and refinement of planting densities. The objective of this study was to provide a theoretical basis for maize production research by analyzing the maize yield gain characteristics. Six varieties of maize were selected for the study; each selection is representative of a typical or commonly used maize variety from a specific decade, beginning from the 1950s and continuing through each decade into the 2000s. The selections and their corresponding decade were as follows: Baihe, 1950s; Jidan 101, 1960s; Zhongdan 2, 1970s; Yedan 13, 1980s; Zhengdan 958, 1990s; and Xianyu 335, 2000s. Each variety was planted under four different densities (37 500, 52 500, 67 500, and 82 500 plants ha-1) and four different nitrogen applications (0, 150, 225, and 300 kg ha-1) to study the effects on yield gain characteristics. The obtained results demonstrated that there was a maize yield increase of 123.19% between the 1950s variety and the 2000s variety. Modern Chinese maize varieties had a higher yield advantage. They also displayed the additional potential to acquire higher yield under increased planting densities and nitrogen applications. At the present cultivation levels (planting at 67 500 plants ha-1 with 225 kg ha-1 nitrogen application), the contribution types and corresponding yield increase percentages were as follows: genetic improvement, 45.37%; agronomic-management improvement, 30.94%; and genotype× agronomicmanagement interaction, 23.69%. At high-yielding cultivation levels (planting at 82 500 plants ha-1 with 300 kg ha-1 nitrogen application), the contribution types and corresponding yield increase percentages were as follows: genetic improvement, 31.30%; agronomic-management improvement, 36.23%; and genotype × agronomic-management interaction, 32.47%. The contribution of agronomic-management and genotype × agronomic-management interaction to yield increase would be larger with the corresponding management improvement. To further increase maize grain yield in China, researchers should further examine the effects of agronomic-management on maize yield and the adaptation of variety to agronomic-management.

Drought is one of the severe meteorological disasters and causes of serious losses for agricultural productions, and early assessment of drought hazard degree is critical in management of maize farming. This study proposes a novel method for assessment of maize drought hazard in different growth stages. First, the study divided the maize growth period into four critical growth stages, including seeding, elongation, tasseling, and filling. Second, maize drought causal factors were selected and the fuzzy membership function was established. Finally, the study built a fuzzy gamma model to assess maize drought hazards, and the gamma 0.93 was finally established using Monte Carlo Analysis. Performing fuzzy gamma operation with 0.93 for gamma and classifying the area yielded a map of maize drought hazards with four zones of light, moderate, severe, and extreme droughts. Using actual field collected data, seven selected samples for drought hazard degree were examined, the model output proved to be a valid tool in the assessment maize drought hazard. This model will be very useful in analyzing the spatial change of maize drought hazard and influence on yield, which is significant for drought management in major agricultural areas.

Experiment was conducted at the Gongzhuling Experimental Station of Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Jilin Province, China, during 2009-2010. Six representative varieties of maize (Baihe in the 1950s, Jidan 101 in the 1960s, Zhongdan 2 in the 1970s, Yedan 13 in the 1980s, Zhengdan 958 in the 1990s, and Xianyu 335 in the 2000s) were each planted under two different densities (52 500 and 82 500 plants ha-1) and two different nitrogen application levels (150 and 300 kg ha-1). Root characteristics and distribution among soil layers were studied by the field root digging method. The results showed that root mass increased with the process of the growth and development of the plant, and it peaked at kernel filling stage, and decreased at maturity due to the root senesces. Root mass of different maize varieties from the 1950s to 1980s had a trend of increase, while it decreased for the modern varieties. Root length and root surface areas had the similar changing trend. The study suggested that early maize varieties may have root redundancy, and reducing root redundancy may be a direction for variety improvement for high yield. Root characteristics were affected by nitrogen application level and density; modern varieties were more suitable for higher fertilizer application level and density conditions. Root characteristics distribution among soil layers decreased by an exponent equation, but the regression coefficients of different varieties were different. Though the root length density (RLD) of every soil layer of different varieties also decreased by an exponent equation, there were large variations of RLD in every part of a layer.

So far, the pathogenesis of demyelination caused by canine distemper virus (CDV) in the central nervous system has remained unclear, although a lot of studies have been done extensively. To further investigate the relation of variety cells in brain to demyelination, this study was performed on 15 dogs with spontaneous acute canine distemper and 2 controls. According to anatomical relation, the brain was divided into cerebrum, cerebral stem and cerebellum. The sections with no, mild, moderate, or severe demyelinating lesions were selected respectively and stained by HE and immunohistochemistry. Immuno-localisation of CDV antigen was used to confirm CDV infection. The brain was examined for co-localisation of the CDV antigen with either an astrocyte-specific marker, glial fibrillary acidic protein (GFAP), or an oligodendrocyte-specific marker, galactocerebroside (GalC). Apoptotic cell was detected by TdT-mediated nick end-labeling assay (TUNEL). The results demonstrated that the local disturbance of blood circulation mainly included congestion, edema, thrombosis, and disseminated intravascular coagulation (DIC). The CDV neucleocapsid protein positive reaction, metabolic disorder and apoptosis of oligodendrocytes were observed in demyelinating areas. Lots of astrocytes displayed CDV antigen-positive, especially in their process. Some of them became apoptotic cell confirmed by TUNEL staining. Fibrous astrocytes showed more intense GFAP-positive in mild and moderate demyelinating area. Some of nervous cells located in pyramidal cell layers and nucleus nervi were in degeneration, necrosis. Satellitosis, neuronophagia and apoptotic neurons were examined by hematoxylin and eosin (HE) and TUNEL staining. The results suggested that the demyelinating changes in brain tissues infected with CDV mainly related to the metabolic disorder and apoptosis of ogliodendrocytes and astrocytes; also involved with the local disturbance of blood circulation and some neuron lost.

Hydrogen peroxide (H2O2) is an important signaling molecule in ethylene-induced stomatal closure in Arabidopsis thaliana. Early studies on the sources of H2O2 mainly focused on NADPH oxidases and cell-wall peroxidases. Here, we report the involvement of polyamine oxidases (PAOs) in ethylene-induced H2O2 production in guard cells. In Arabidopsis epidermal peels, application of PAO inhibitors caused the failure of ethylene to induce H2O2 production and stomatal closure. Results of quantitative RT-PCR analysis and pharmacological experiments showed that AtPAO2 and AtPAO4 transcripts and activities of PAOs were both induced by ethylene. In transgenic Arabidopsis plants over-expressing AtPAO2 and AtPAO4, stomatal movement was more sensitive to ethylene treatment and H2O2 production was also significantly induced. The increased H2O2 production in the transgenic lines compared to the wild-type plants suggests that AtPAO2 and AtPAO4 probably are involved in ethylene-induced H2O2 production. Several factors which induce stomatal closure such as dehydration and high salinity all enhanced the expression of AtPAO2 and AtPAO4 to different degrees. Moreover, GFPAtPAOs fusion protein localized in the nucleus, cytoplasm, and cell wall of the guard cells. Therefore, our results strongly indicated that PAO is a source of H2O2 generation in Arabidopsis guard cells and plays crucial roles in stomatal movement.

2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA), the dominant benzoxazinoid hydroxamic acid in maize (Zea Mays L.), serves as important factors of resistance against insects and microbial diseases, allelochemicals used in competition with other plants. In this paper, a novel and simple method for the isolation and purification of DIMBOA from maize seedlings was developed. Frozen shoots from 7-d-old maize seedlings (1 000×g) were firstly defrosted and then were directly homogenized and extracted with ethyl acetate. The macerate was allowed to stand at room temperature (25±2)°C for 1 h to allow enzymatic release of DIMBOA from DIMBOA-glucoside. Then the ethyl acetate phase was filtered, dried and evaporated to dryness. The resulting light-tan, semicrystalline residue was stored at -20°C for 24 h. Upon recrystallization from acetone-hexane, a relative higher yield (0.58 g) of pure DIMBOA crystals was obtained compared with the yield afforded by Woodward methodology (0.26 g).

Pharmacological, laser scanning confocal microscopic (LSCM), and spectrophotographic approaches were used to study the roles of hydrogen sulfide (H2S) and nitric oxide (NO) in signaling transduction of stomatal movement in response to ethylene in Vicia faba L. Ethylene treatment resulted in the dose-dependent stomatal closure under light, and this effect was blocked by the inhibitors of H2S biosynthesis in V. faba L. Additionally, ethylene induces H2S generation and increases L-/D-cysteine desulfhydrase (pyridoxalphosphate-dependent enzyme) activity in leaves of V. faba L. Inhibitors of H2S biosynthesis have no effect on the ethylene-induced stomatal closure, NO accumulation, and nitrate reductase (NR) activity in guard cells or leaves of V. faba L. Moreover, the ethylene-induced increase of H2S levels and L-/Dcysteine desulfhydrase activity declined when NO generation was inhibited. Therefore, we conclude that H2S and NO probably are involved in the signal transduction pathway of ethylene-induced stomatal closure. H2S may represent a novel component downstream of NO in the ethylene-induced stomatal movement in V. faba L.