The identification of stable quantitative trait locus (QTL) for yield-related traits and tightly linked molecular markers is important for improving wheat grain yield. In the present study, six yield-related traits in a recombinant inbred line (RIL) population derived from the Zhongmai 578/Jimai 22 cross were phenotyped in five environments. The parents and 262 RILs were genotyped using the wheat 50K single nucleotide polymorphism (SNP) array. A high-density genetic map was constructed with 1 501 non-redundant bin markers, spanning 2 384.95 cM. Fifty-three QTLs for six yield-related traits were mapped on chromosomes 1D (2), 2A (9), 2B (6), 2D, 3A (2), 3B (2), 4A (5), 4D, 5B (8), 5D (2), 7A (7), 7B (3) and 7D (5), which explained 2.7–25.5% of the phenotypic variances. Among the 53 QTLs, 23 were detected in at least three environments, including seven for thousand-kernel weight (TKW), four for kernel length (KL), four for kernel width (KW), three for average grain filling rate (GFR), one for kernel number per spike (KNS) and four for plant height (PH). The stable QTLs QKl.caas-2A.1, QKl.caas-7D, QKw.caas-7D, QGfr.caas-2B.1, QGfr.caas-4A, QGfr.caas-7A and QPh. caas-2A.1 are likely to be new loci. Six QTL-rich regions on 2A, 2B, 4A, 5B, 7A and 7D, showed pleiotropic effects on various yield traits. TaSus2-2B and WAPO-A1 are potential candidate genes for the pleiotropic regions on 2B and 7A, respectively. The pleiotropic QTL on 7D for TKW, KL, KW and PH was verified in a natural population. The results of this study enrich our knowledge of the genetic basis underlying yield-related traits and provide molecular markers for high-yield wheat breeding.

SrUGT76G1, the most well-studied diterpene glycosyltransferase in Stevia rebaudiana, is key to the biosynthesis of economically important steviol glycosides (SGs).  However, the molecular regulatory mechanism of SrUGT76G1 has rarely been explored.  In this study, we identified a MYB transcription factor, SrMYB1, using a yeast one-hybrid screening assay.  SrMYB1 belongs to the typical R2R3-type MYB protein and is specifically localized in the nucleus with strong transactivation activity.  The transcript of SrMYB1 is predominantly accumulated in flowers, but is also present at a lower level in leaves.  Yeast one-hybrid and electrophoretic mobility shift assays verified that SrMYB1 binds directly to the MYB binding sites in the F4-3 fragment (+50–(–141)) of the SrUGT76G1 promoter.  Furthermore, we found that SrMYB1 could significantly repress the expression of SrUGT76G1 in both epidermal cells of tobacco leaves and stevia callus.  Taken together, our results demonstrate that SrMYB1 is an essential upstream regulator of SrUGT76G1 and provide novel insight into the regulatory network for the SGs metabolic pathway in S. rebaudiana.

Resistant cultivar deployment is an effective method for cereal aphid management.  Under greenhouse conditions, preliminary antibiosis resistance screening was conducted on 114 Ethiopian and 22 Chinese spring wheat accessions.  After performing a bioassay to determine antibiosis resistance, aphid feeding behaviour and phenolic acid content analyses were performed on the aphid resistant wheat accessions by electrical penetration graph (EPG) and high performance liquid chromatography (HPLC), respectively.  Among the wheat accessions, two high resistances, 27 moderate-resistances, and 35 low-resistances to Sitobion miscanthi were identified.  The antibiosis resistance test showed prolonged pre-adult and pre-reproductive periods, shorter reproductive periods, lower fecundity, an intrinsic rate (r_m) of increase, and a finite rate (λ) of increase of S. miscanthi on Lunxuan 145, Wane, Lunxuan 6, 204511, Lunxuan 103 and 5215 than those on the aphid-susceptible accession Beijing 837.  The changes for the parameters of aphid feeding behaviour, including spending a longer time in the penetration and phloem salivation phases and less time in the phloem sap-feeding phase on the resistant wheat accessions, the aphid resistance may occur during the phloem phase and may be due to mechanicalAdditionally, the HPLC analysis showed higher contents of: 1) ferulic acid in Lunxuan 145, Lunxuan 103 and Lunxuan 6; 2) p-coumaric acid in Lunxuan145; 3) vanillic acid in Lunxuan 145, Wane and Lunxuan 6; 4) syringic acid in Lunxuan 103; and 5) caffeic acid in 5215.  The contents of some phenolic acids within wheat leaves, such as p-courmaric acid and vanillic acid showed significant positive correlation with the duration of aphid development, but negative correlation with the aphid fecundity.  The concentrations of these acids may be the causes of antibiosis resistance to S. miscanthi.  The identification of grain aphid-resistant wheat accessions in our study will be helpful in future breeding program for pest control.

Soluble sugar content in seeds is an important quality trait of soybean.  In this study, 57 quantitative trait loci (QTLs) related to soluble sugar contents in soybean seeds were collected from databases and published papers.  After meta-overview-collinearity integrated analysis to refine QTL intervals, eight consensus QTLs were identified.  To further verify the consensus QTLs, a population of chromosome segment substitution lines (CSSLs) was analyzed.  Two lines containing fragments covering the regions of consensus QTLs and the recurrent parent were selected: one line showed high soluble sugar contents associated with a consensus QTL fragment, and the other line showed low soluble sugar contents.  Transcriptome sequencing was conducted for these two lines at the early, middle, and late stages of seed development, which identified 158, 109 and 329 differentially expressed genes, respectively.  Based on the analyses of re-sequencing data of the CSSLs and the consensus QTL region, three candidate genes (Glyma.19G146800, Glyma.19G122500, and Glyma.19G128500) were identified in the genetic fragments introduced from wild soybean.  Sequence comparisons between the two CSSL parents SN14 and ZYD00006 revealed a single nucleotide polymorphism (SNP) mutation in the coding sequence of Glyma.19G122500, causing a non-synonymous mutation in the amino acid sequence that affected the predicted protein structure.  A Kompetitive allele-specific PCR (KASP) marker was developed based on this SNP and used to evaluate the CSSLs.  These results lay the foundation for further research to identify genes related to soluble sugar contents in soybean seeds and for future soybean breeding.

Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.  Volatiles emitted from A. annua attract A. lucorum.  Volatile artemisinic acid of A. annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field.  However, little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A. lucorum.  In this study, we collected volatiles from A. annua at the seedling stage by using headspace solid phase microextraction (HS-SPME).  Gas chromatography-mass spectrometry (GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng h^–1 artemisinic acid were detected in volatile samples and milled samples, respectively.  Subsequently, a key gene for artemisinic acid synthesis, the cytochrome P450 gene cyp71av1, was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid.  After the addition of exogenous artemisinic alcohol or artemisinic aldehyde, artemisinic acid was identified as the product of the expressed gene.  In electroantennogram (EAG) recordings, 3-day-old adult A. lucorum showed significant electrophysiological responses to artemisinic alcohol, artemisinic aldehyde and artemisinic acid.  Furthermore, 3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L^–1, whereas 3-day-old male bugs were attracted significantly by 10 mmol L^–1 artemisinic acid and artemisinic aldehyde.  We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A. lucorum.

The wheat aphid, Sitobion miscanthi, is one of the most destructive pests of wheat plants in the temperate regions of China.  Little is known about the genetic structure evolution of  the different geographic populations of S. miscanthi with its migration.  In this study, we investigated the population genetic structure and demographic history of S. miscanthi by analysing 18 geographical populations across China using one mitochondrial gene, COI; one nuclear gene, EF-1α; and two endosymbiont Buchnera genes, gnd and trpA.  Analysis of data from the various groups showed high haplotype diversity and low nucleotide variation.  SAMOVA analysis did not find a correlation between genetic distance and geographic distance.  However, areas with high population diversity exhibited high haplotype diversity.  Therefore, we speculate that there are two main natural migration pathways of S. miscanthi in China.  One is from Yunnan to the Sichuan Basin, and the other is from Wuhan, Xinyang and Jiaodong Peninsula areas to the northwest.  Based on this hypothesis, we inferred that these aphid populations appear first in the southwestern and southern regions and spread to the north with the help of the southeastern and southwestern monsoons, which occur in spring and summer.  In autumn, the aphids spread southward with the northeastern and northwestern monsoons.

Topping is a cultivation method that is widely practiced due to the indeterminate growth character of cotton (Gossypium hirsutum L.).  Among the different methods of accomplishing topping, manual topping is common in the Yellow River Valley of China, although it is time- and labor-intensive.  The objective of this study was to characterize the responses of cotton to different topping treatments with respect to development, yield and quality.  This study included field experiments from 2015 to 2016 with three different topping methods: manual topping (MT), chemical topping (CT) using mepiquat chloride, and a non-decapitation treatment (NT).  We found that the plant height, the number of fruiting branches and the length of upper fruiting branches of cotton treated with CT were significantly lower than NT.  The chlorophyll content of cotton treated with CT was not significantly different from NT, but was higher than that of MT in the later season.  CT enhanced plant development with reduced endogenous gibberellic acid and abscisic acid contents, and the apical development of the main stem was inhibited.  Compared with MT, CT significantly increased the biomass of the vegetative parts.  Most importantly, there were no significant differences in the yield or fiber quality between MT and CT.  These findings suggested that CT, a simplified and effective topping method, could be utilized as an alternative in the Yellow River Valley of China.

The objective of this work was to develop a model for simulating the leaf color dynamics of winter wheat in relation to crop growth stages and leaf positions under different nitrogen (N) rates.  RGB (red, green and blue) data of each main stem leaf were collected throughout two crop growing seasons for two winter wheat cultivars under different N rates.  A color model for simulating the leaf color dynamics of winter wheat was developed using the collected RGB values.  The results indicated that leaf color changes went through three distinct stages, including early development stage (ES), early maturity stage (MS) and early senescence stage (SS), with respective color characteristics of light green, dark green and yellow for the three stages.  In the ES stage, the R and G colors gradually decreased from their initial values to steady values, but the B value generally remained unchanged.  RGB values remained steady in the MS, but all three gradually increased to steady values in the SS.  Different linear functions were used to simulate the dynamics of RGB values in time and space.  A cultivar parameter of leaf color matrix (M_RGB) and a nitrogen impact factor (FN) were added to the color model to quantify their respective effects.  The model was validated with an independent experimental dataset.  RMSEs (root mean square errors) between the observed and simulated RGB values ranged between 7.0 and 10.0, and relative RMSEs (RRMSEs) ranged between 7 and 9%.  In addition, the model was used to render wheat leaves in three-dimensional space (3D).  The 3D visualizations of leaves were in good agreement with the observed leaf color dynamics in winter wheat.  The developed color model could provide a solid foundation for simulating dynamic crop growth and development in space and time. 

Understanding yield potential, yield gap and the priority of management factors for reducing the yield gap in current intensive maize production is essential for meeting future food demand with the limited resources. In this study, we conducted field experiments using different planting modes, which were basic productivity (CK), farmer practice (FP), high yield and high efficiency (HH), and super high yield (SH), to estimate the yield gap. Different factorial experiments (fertilizer, planting density, hybrids, and irrigation) were also conducted to evaluate the priority of individual management factors for reducing the yield gap between the different planting modes. We found significant differences between the maize yields of different planting modes. The treatments of CK, FP, HH, and SH achieved 54.26, 58.76, 65.77, and 71.99% of the yield potential, respectively. The yield gaps between three pairs: CK and FP, FP and HH, and HH and SH, were 0.76, 1.23 and 0.85 t ha^–1, respectively. By further analyzing the priority of management factors for reducing the yield gap between FP and HH, as well as HH and SH, we found that the priorities of the management factors (contribution rates) were plant density (13.29%)>fertilizer (11.95%)>hybrids (8.19%)>irrigation (4%) for FP to HH, and hybrids (8.94%)>plant density (4.84%)>fertilizer (1.91%) for HH to SH. Therefore, increasing the planting density of FP was the key factor for decreasing the yield gap between FP and HH, while choosing hybrids with density and lodging tolerance was the key factor for decreasing the yield gap between HH and SH.

Abiotic stress poses a great threat to plant growth and can lead to huge losses in yield.  Gene enolase2 (ENO2) is important in resistance to abiotic stress in various organisms.  ENO2 T-DNA insertion mutant (eno2^–) plants of Arabidopsis thaliana showed complete susceptibility to sodium chloride treatment when were analyzed either as whole plants or by measuring root growth during NaCl treatment.  Quantitative real-time RT-PCR (RT-qPCR) was performed to investigate the expression profile of ENO2 in response to NaCl stress in Arabidopsis.  The transcript level of ENO2 was rapidly elevated in 300 mmol L^–1 NaCl treatment.  ENO2 also responded to 300 mmol L^–1 NaCl treatment at the protein level.  To illuminate the mechanism underlying ENO2 resistance to salt at the transcriptional level, we studied the wild-type and eno2^– Arabidopsis lines that were treated with 300 mmol L^–1 NaCl for 18 h using 454 GS FLX, which resulted in an expressed sequence tag (EST) dataset.  A total of 961 up-regulated and 746 down-regulated differentially expressed genes (DEGs) were identified in the pairwise comparison WT-18 h:eno2^–-18 h.  The DEGs were identified and functionally annotated using the databases of Gene Ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG).  The identified unigenes were subjected to GO analysis to determine biological, molecular, and cellular functions.  The biological process was enriched in a total of 20 GO terms, the cellular component was enriched in 13 GO terms, and the molecular function was enriched in 11 GO terms.  Using KEGG mapping, DEGs with pathway annotations contributed to 115 pathways.  The top 3 pathways based on a statistical analysis were biosynthesis of the secondary metabolites (KO01110), plant-pathogen interactions (KO04626), and plant hormone signal transduction (KO04075).  Based on these results, ENO2 contributes to increased resistance to abiotic stress.  In particular, ENO2 is involved in some of the metabolic stress response pathways in Arabidopsis.  Our work also demonstrates that this EST dataset will be a powerful resource for further studies of ENO2, such as functional analyses, investigations of biological roles, and molecular breeding.  Additionally, 3-phosphoglycerate kinase (PGK), 3-phosphoglycerate kinase 1 (PGK1), triosephosphate isomerase (TPI), and pyruvate kinase (PK) in glycolysis interactions with ENO2 were verified using the yeast two-hybrid experiment, and ENO2 may regulate the expression of PGK, PGK1, TPI, and PK.  Taken together, the results from this study reflects that ENO2 gene has an important role in the response to the high salt stress.

The objective of this work was to develop a dynamic model for describing leaf curves and a detailed spatial geometry model of the rice leaf (including sub-models for unexpanded leaf blades, expanded leaf blades, and leaf sheaths), and to realize three-dimensional (3D) dynamic visualization of rice leaves by combining relevant models.  Based on the experimental data of different cultivars and nitrogen (N) rates, the time-course spatial data of leaf curves on the main stem were collected during the rice development stage, then a dynamic model of the rice leaf curve was developed using quantitative modeling technology.  Further, a detailed 3D geometric model of rice leaves was built based on the spatial geometry technique and the non-uniform rational B-spline (NURBS) method.  Validating the rice leaf curve model with independent field experiment data showed that the average distances between observed and predicted curves were less than 0.89 and 1.20 cm at the tilling and jointing stages, respectively.  The proposed leaf curve model and leaf spatial geometry model together with the relevant previous models were used to simulate the spatial morphology and the color dynamics of a single leaf and of leaves on the rice plant after different growing days by 3D visualization technology.  The validation of the leaf curve model and the results of leaf 3D visualization indicated that our leaf curve model and leaf spatial geometry model could efficiently predict the dynamics of rice leaf spatial morphology during leaf development stages.  These results provide a technical support for related research on virtual rice.

    Flixweed (Descurainia sophia L.) is a problematic and widespread weed in winter wheat fields and has been controlled by tribenuron-methyl for more than twenty years in China. In this study, a flixweed accession (Hebei 25, HB25) with an Asp-376-Glu mutation in acetohydroxy acid synthase (AHAS) was identified and purified. The purified HB25 accession (pHB25) developed 758.1-fold resistance to tribenuron-methyl and exhibited obvious cross-resistance to four AHAS-inhibiting herbicides. The resistant/susceptible (R/S) ratios of 50% plant growth reduction (GR₅₀) to herbicides of halosulfuron-methyl, flumetsulam, imazethapyr and pyribenzoxim were 346.1, 15.7, 8.1 and 7.1, respectively. The reduced AHAS sensitivities to four different AHAS-inhibiting herbicides, which were caused by the Asp-376-Glu mutation, were responsible for the resistance and cross-resistance to AHAS-inhibiting herbicides. The R/S ratios of 50% inhibition of AHAS activity (I₅₀) to tribenuron-methyl, halosulfuron-methyl, flumetsulam, imazethapyr and pyribenzoxim were 844.5, 532.9, 74.5, 13.3 and 5.5, respectively. The results of AHAS activity in vitro were highly correlated with that of whole-plant response experiments.

The Citrus tristeza virus (CTV) uses 3 silencing suppressor genes, p20, p23 and p25, to resist the attacks from its Citrus hosts. Inactivating these genes is therefore obviously a potential defensive option in addition to the current control strategies including aphid management and the use of mild strain cross protection. In this study, we cloned partial DNA fragments from the three genes, and used them to construct vectors for expressing hairpin RNAs (hpRNAs). To facilitate the formation of hpRNAs, the constructs were introduced in a loop structure. Following transformation of sour orange (Citrus aurantium) with these constructs, 8 p20 hpRNA (hp20) and 1 p25 hpRNA (hp25) expressing lines were obtained. The 7 hp20 transgenic lines were further characterized. Their reactions to CTV were tested following inoculation with CT14A and/ or TR-L514, both of which are severe strains. Results showed that 3 lines (hp20-5, hp20-6 and hp20-8) were completely resistant to TR-L514 under greenhouse conditions for no detectable viral load was found in their leaves by PCR. However, they exhibited only partial suppression of TR-L514 under screen house conditions since the virus was detected in their leaves, though 2 months later compared to non-transgenic controls. Further tests showed that hp20-5 was tolerant also to CT14A under screen house conditions. The growth of hp20-5 was much better than others including the controls that were concurrently challenged with CT14A. These results showed that expressing p20 hpRNA was sufficient to confer sour orange with CTV resistance/tolerance.

A real-time RT-PCR assay using TaqMan-MGB probes was developed to detect and type the bovine viral diarrhea virus (BVDV) in cattle. Universal primers and TaqMan-MGB probes were designed from the 5´-untranslated region of known pestiviral sequences. Prior to optimizing the assay, cRNAs were transcribed in vitro from the BVDV 1 and BVDV 2 RTPCR products to make standard curves. The detection limit of the assay was 1.72×102 copies for BVDV 1 and 2.14×102 copies for BVDV 2. The specificity of the assay evaluated on several BVDV strains including bovine herpesvirus 1 (BHV 1), foot and mouth disease virus (FMDV) and several classical swine fever virus (CSFV) strains showed specific detection of the positive virus over 40 cycles. The assay was highly reproducible with the coefficient of variance ranging from 1.04 to 1.33% for BVDV 1 and from 0.83 to 1.48% for BVDV 2, respectively. Using this method, we tested a total of 2 327 cattle from three dairy farms for the presence of BVDV persistently infected (PI) animals. In this assay, each RT-PCR template contained a mixture of ten samples from different animals. The occurrence rate of PI cattle in three farms ranging from 0.9 to 2.54% could represent partly the PI rates in cattle farm in China. In conclusion, using our real-time PCR assay, we could effectively detect and type BVDV and identify PI cattle in a rapid and cost-effective manner.

MicroRNAs (miRNAs) are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal. To discover novel miRNAs and identify the differential expression of them in ovine ovary and testis tissues, the study constructed two libraries by using next-generation sequencing technologies (Solexa high-throughput sequencing technique). As a result, 9 321 775 and 9 511 538 clean reads were obtained from the ovary and testis separately, which included 130 562 (90 genes of ovary) and 56 272 (85 genes of testis) of known miRNAs and 486 potential novel miRNAs reads. In this study, a total of 65 conserved miRNAs were significantly differentially expressed (P<0.01) between the two samples. Among them, 28 miRNAs were up-regulated and 3 miRNAs were down-regulated on ovary compared with testis. In addition, the known miRNAs with the highest expression level (5 miRNAs) and 30 novel miRNAs with the functions related to reproduction were validated using the real-time quantitative RT-PCR. Moreover, the gene ontology (GO) annotation and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that differentially expressed miRNAs were involved in ovary and testis physiology, including signal transduction, gonad development, sex differentiation, gematogenesis, fertilization and embryo development. The results will be helpful to facilitate studies on the regulation of miRNAs during ruminant reproduction.

The full-length cDNA of functionally-unknown salivary protein C002 in Schizaphis graminum was cloned using rapid amplification of cDNA ends (RACE) and designated as SgC002 (GenBank accession no. KC977563). It is 767 bp long and encodes a protein of 190 amino acid residues with a predicted mass of 21.5 kDa and a predicted cleavage site of N-terminal signal peptide between the 24th and the 25th residues. SgC002 is specifically expressed in salivary gland with the highest level at the 2nd instar. Introducing SgC002-specific 476-siRNA, but not 546-siRNA to aphids through artificial diet significantly suppressed SgC002 expression. Silencing SgC002 gene led to lethality of the aphid on wheat plants, but not on pure artificial diet. Our study demonstrated that artificial diet-mediated RNAi can be a useful tool for research on the roles of genes in aphid salivary gland, and also provided new insights into the characteristics of C002 in wheat aphids.

This paper was to develop a model for simulating the leaf color changes in rice (Oryza sativa L.) based on RGB (red, green, and blue) values. Based on rice experiment data with different cultivars and nitrogen (N) rates, the time-course RGB values of each leaf on main stem were collected during the growth period in rice, and a model for simulating the dynamics of leaf color in rice was then developed using quantitative modeling technology. The results showed that the RGB values of leaf color gradually decreased from the initial values (light green) to the steady values (green) during the first stage, remained the steady values (green) during the second stage, then gradually increased to the final values (from green to yellow) during the third stage. The decreasing linear functions, constant functions and increasing linear functions were used to simulate the changes in RGB values of leaf color at the first, second and third stages with growing degree days (GDD), respectively; two cultivar parameters, MatRGB (leaf color matrix) and AR (a vector composed of the ratio of the cumulative GDD of each stage during color change process of leaf n to that during leaf n drawn under adequate N status), were introduced to quantify the genetic characters in RGB values of leaf color and in durations of different stages during leaf color change, respectively; FN (N impact factor) was used to quantify the effects of N levels on RGB values of leaf color and on durations of different stages during leaf color change; linear functions were applied to simulate the changes in leaf color along the leaf midvein direction during leaf development process. Validation of the models with the independent experiment dataset exhibited that the root mean square errors (RMSE) between the observed and simulated RGB values were among 8 to 13, the relative RMSE (RRMSE) were among 8 to 10%, the mean absolute differences (da) were among 3.85 to 6.90, and the ratio of da to the mean observation values (dap) were among 3.04 to 4.90%. In addition, the leaf color model was used to render the leaf color change over growth progress using the technology of visualization, with a good performance on predicting dynamic changes in rice leaf color. These results would provide a technical support for further developing virtual plant during rice growth and development.

Somatic hybridization is performed to obtain significant cytoplasmic male sterility (CMS) lines, whose CMS genes are derived either from the transfer of sterile genes from the mitochondrial genome of donor parent to the counterpart of receptor or production of new sterile genes caused by mitochondrial genome recombination of the biparent during protoplast fusion. In this study, a novel male sterile line, SaNa-1A, was obtained from the somatic hybridization between Brassica napus and Sinapis alba. The normal anther development of the maintainer line, SaNa-1B, and the abortive process of SaNa-1A were described through phenotypic observations and microtome sections. The floral organ of the sterile line SaNa-1A was sterile with a shortened filament and deflated anther. No detectable pollen grains were found on the surface of the sterile anthers. Semi-thin sections indicated that SaNa-1A aborted in the pollen mother cell (PMC) stage when vacuolization of the tapetum and PMCs began. The tapetum radically elongated and became highly vacuolated, occupying the entire locule together with the vacuolated microspores. Therefore, SaNa-1A is different from other CMS lines, such as ogu CMS, pol CMS and nap CMS as shown by the abortive process of the anther.

Intramuscular fat (IMF) content is considered to be a key factor that affects the marbling, tenderness, juiciness and flavor of pork. To investigate the effects of myristic acid (MA) on the differentiation of porcine intramuscular adipocytes, cells were isolated from longissimus dorsi muscle (LDM) and treated with 0, 10, 50 or 100 μmol L-1 MA. The results showed that MA significantly promotes the differentiation of intramuscular adipocytes in a dose-dependent manner. MA also led to a parallel increase in the expression of peroxisome proliferator activated receptor-γ (PPARγ) and adipose-related genes, such as glucose transporter 1 (GLUT1), lipoprotein lipase (LPL), adipocyte fatty acid binding protein 4 (FABP4/aP2), fatty acid translocase (FAT), acetyl-CoA carboxylase α (ACCα), adipose triglyceride lipase (ATGL) and fatty acid synthase (FASN). However, no significant effects of MA were observed on the expression of CAAT enhancer binding protein-α (C/EBPα) or hormone sensitive lipase (HSL). The expression of pyruvate dehydrogenase kinase 4 (PDK4) was increased by MA during the early stages of differentiation (day 1-3). In addition, MA also increased the absolute content of C14 (P<0.001) and saturated fatty acids (SFA) (P<0.05) to varying degrees, but no effects were observed on other fatty acids. These results suggest that MA might be able to enhance the IMF content of pork and increase the accumulation of myristic and myristoleic acid in muscle, which might have beneficial implications for human health.

Water shortage is a serious issue threatening the sustainable development of agriculture in the North China Plain, with the winter wheat (Triticum aestivum L.) as its largest water-consuming crop. The effects of tillage practices on the water consumption and water use efficiency (WUE) of wheat under high-yield conditions using supplemental irrigation based on testing soil moisture dynamic change were examined in this study. This experiment was conducted from 2007 to 2010, with five tillage practice treatments, namely, strip rotary tillage (SR), strip rotary tillage after subsoiling (SRS), rotary tillage (R), rotary tillage after subsoiling (RS), and plowing tillage (P). The results showed that in the SRS and RS treatments the total water and soil water consumptions were 11.81, 25.18% and 12.16, 14.75% higher than those in SR and R treatments, respectively. The lowest ratio of irrigation consumption to total water consumption in the SRS treatment was 18.53 and 21.88% for the 2008-2009 and 2009- 2010 growing seasons, respectively. However, the highest percentage of water consumption was found in the SRS treatment from anthesis to maturity. No significant difference was found between the WUE of the flag leaf at the later filling stage in the SRS and RS treatments, but the flag leaf WUE at these stages were higher than those of other treatments. The SRS and RS treatments exhibited the highest grain yield (9 573.76 and 9 507.49 kg ha-1 for 3-yr average) with no significant difference between the two treatments, followed by P, R and SR treatments. But the SRS treatment had the highest WUE. Thus, the 1-yr subsoiling tillage, plus 2 yr of strip rotary planting operation may be an efficient measure to increase wheat yield and WUE.

This study is first to investigate proteomic changes in sheep sperm induced by carbon ion radiation using two-dimensional electrophoresis (2-DE) analysis in the project of breeding a new variety of sheep. Differential expression proteins were detected using the PDQuest 8.0 software after staining with Coomassie blue. Valid spots were then analyzed through liquid chromatography tandem mass spectrometry (LC-MS/MS). Among the 480 total protein spots displayed in 2-D gels, 6 specific protein spots were observed in sperm gels. A search against protein sequences in the National Center for Biotechnology Information databases (NCBI) indicated that differentially expressed proteins correspond to two proteins, identified to be enolase and transcription factor AP-2-alpha (TFAP-2α). The two proteins were up-regulated in the irradiated sperm. To the best of our knowledge, this study is the first to identify proteomic changes induced by carbon ion radiation in sheep sperm. The analysis of differential expression protein may be useful in identifying new breeding markers in sheep reproduction and in clarifying the mechanisms involved in irradiation or space breeding.

Cry toxins produced by Bacillus thuringiensis (Bt) are effective biological insecticides against certain insect species. However, there are potential risks of the evolved resistance of insects to Cry toxin owing to decreased binding of toxins to target sites in the brush border membranes of the larva midgut. The Cry toxins with different binding sites in the larval midgut have been considered to be a good combination to deploy in delaying resistance evolution. Bioassay results demonstrated that the toxicity of different Cry toxins ranked differently for each species. The toxicity ranking was Cry1Ac>Cry1Ab>Cry2Ab for Helicoverpa armigera, Cry1B>Cry1C>Cry2Ab for Spodoptera exigua, and Cry2Ab>Cry1B> Cry1C for S. litura. Only Cry2Ab was toxic to Agrotis ipsilon. Binding experiments were performed with 125I-Cry1Ab, 125ICry1Ac, 125I-Cry1B, 125I-Cry1C, 125I-Cry2Ab and the brush border membranes vesicles (BBMV) from H. armigera, S. exigua, S. litura and A. ipsilon. The binding of Cry1Ab and Cry1Ac was shown to be saturable by incubating with increasing concentrations of H. armigera BBMV (Kd=(45.00±2.01) nmol L-1 and (12.80±0.18) nmol L-1, respectively; Bmax=(54.95±1.79) ng and (55.44±0.91) ng, separately). The binding of Cry1B was shown to be saturable by incubating with increasing concentrations of S. exigua BBMV (Kd=(23.26±1.66) nmol L-1; Bmax=(65.37±1.87) ng). The binding of 125ICry toxins was shown to be non-saturable by incubating with increasing concentrations of S. litura and A. ipsilon BBMV. In contrast, Cry1B and Cry1C showed some combination with the BBMV of S. litura, and a certain amount of Cry2Ab could bind to the BBMV of A. ipsilon. These observations suggest that a future strategy could be devised for the focused combination of specific cry genes in transgenic crops to control target pests, widen the spectrum of insecticide effectiveness and postpone insect resistance evolution.

A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera (Hübner) was obtained from antennal cDNA libraries and expressed in Escherichia coli. The real time quantitative PCR (RT-qPCR) results indicated that HarmCSP5 gene was mainly expressed in male and female antennae but also expressed in female legs and wings. Competitive binding assays were performed to test the binding affinity of recombinant HarmCSP5 to 60 odor molecules including some cotton volatiles. The resules showed that HarmCSP5 showed strong binding abilities to 4-ehtylbenzaldehyde and 3,4-dimethlbenz aldehyde, whereas methyl phenylacetate, 2-decanone, 1-pentanol, carvenol, isoborneol, nerolidol, 2- nonanone and ethyl heptanoate have relatively weak binding affinity. Moreover, the predicted 3D model of HarmCSP5 consists of six α-helices located among residues 33-38 (α1), 40-48 (α2), 62-72 (α3), 80-96 (α4), 98-108 (α5), and 116-119 (α6), two pairs of disulfide bridges Cys49-Cys55, Cys75-Cys78. The two amino acid residues, Ile94 and Trp101, may play crucial roles in HarmCSP5 binding with ligands and need further study for confirmation.

The Italian wheat cv. Strampelli displays high resistance to powdery mildew caused by Blumeria graminis f. sp. tritici. The objective of this study was to map quantitative trait loci (QTLs) for resistance to powdery mildew in a population of 249 F2:3 lines from Strampelli/Huixianhong. Adult plant powdery mildew tests were conducted over 2 yr in Beijing and 1 yr in Anyang and simple sequence repeat (SSR) markers were used for genotyping. QTLs Qpm.caas-3BS, Qpm.caas-5BL.1, and Qpm.caas-7DS were consistent across environments whereas, Qpm.caas-2BS.1 found in two environments, explained 0.4-1.6, 5.5-6.9, 27.1-34.5, and 1.0-3.5% of the phenotypic variation respectively. Qpm.caas-7DS corresponded to the genomic location of Pm38/Lr34/Yr18. Qpm.caas-4BL was identified in Anyang 2010 and Beijing 2011, accounting for 1.9-3.5% of phenotypic variation. Qpm.caas-2BS.1 and Qpm.caas-5BL.1 contributed by Strampelli and Qpm.caas-3BS by Huixianhong, seem to be new QTL for powdery mildew resistance. Qpm.caas-4BL, Qpm.caas-5BL.3, and Qpm.caas-7DS contributed by Strampelli appeared to be in the same genomic regions as those mapped previously for stripe rust resistance in the same population, indicating that these loci conferred resistance to both stripe rust and powdery mildew. Strampelli could be a valuable genetic resource for improving durable resistance to both powdery mildew and stripe rust in wheat.

In recent years, subgroup J avian leukosis virus (ALV-J) has been found to frequently infect layers in China. This virus is responsible for economic losses due to both mortality and decreased performance in chickens. In this study, 45-d-old cloned free-range layers were suspected to be infected with ALV and other immunosuppressive diseases because their feathers were unkempt and their growth rate was impaired. To estimate the infection status and determine the source of ALV-J in the flock, 30 cloacal swabs were randomly collected to measure the p27 antigen level by enzyme-linked immunosorbent assay (ELISA). Among the birds that were tested, 87% (26/30) were positive. In addition, 6 anticoagulant blood samples were aseptically collected at random from the flock when the layers were 60 d old. These samples were centrifuged to obtain the leukocytes, which were then used to inoculate chicken embryo fibroblast (CEF) cells for the identification of ALV-J by indirect immunofluorescence (IFA). Of the samples tested, 100% (6/6) were positive. The flock’s production performance was also investigated, and 10 layers were necropsied to evaluate pathological changes at 115 d of age. The flock never laid eggs even though they reached the age of the first laying (110 d). Furthermore, there were pathological changes present, including atrophy of the thymus and bursa of Fabricius, undeveloped ovaries, glandular stomach haemorrhage, and hepatosplenomegaly. Paraffin-embedded sections of intumescent liver and spleen were prepared for antigen localisation using IFA. Positive signals were prevalent in paraffin-embedded sections of the intumescent liver and spleen. Furthermore, provirus DNA was extracted from 4 cloned free-range layers, and 2 paternal parents (HR native cocks), and the gp85 gene of ALV-J was amplified by PCR to analyse the genetic variation. The results of the autogenous variation analysis showed that the 6 strains were 98.5-99.7% homologous. This study indicated that there was persistent infection with ALV-J by dynamic inspection, which seriously reduced the production performance of the flock. In addition, the genetic variation analysis showed that ALV-J in the flock was more likely to have originated from the paternal parent, the HR native cock.

To evaluate the possible genetic interrelationships between flour components and the sedimentation volume (SD), a doubled haploid (DH) population comprising 168 lines were used to identify the conditional quantitative trait loci (QTLs) for SD in three environments. Ten additive QTLs and 15 pairs of epistatic QTLs were detected for SD through unconditional and conditional QTL mapping. Three major additive QTLs were detected for SD conditioned on the seven quality traits. Two additive QTLs were found to be independent of these traits. Three additive QTLs were suppressed by three of the seven traits because of non-detection in unconditional mapping. Three pairs of epistatic QTLs were completely affected by the seven traits because of detection in unconditional mapping but no-detection in conditional mapping. Twelve pairs of epistatic QTLs were detected in conditional mapping. Our results indicated that conditional mapping could contribute to a better understanding of the interdependence of different and closely correlated traits at the QTL molecular level, especially some minor QTLs were found. The conditional mapping approach provides new insights that will make it possible to avoid the disadvantages of different traits by breeding through molecular design.