Identifying the sex pheromone systems of local pest populations facilitates their management, especially for moth species that show significant geographic variation in sex pheromone communication.  We investigated the pheromone production and behavioral responses of the Asian corn borer (Ostrinia furnacalis Guenée; ACB) in Xinjiang, China.  The ACB produces three compounds: (Z)-12-tetradecenyl acetate (Z12-14:Ac) and (E)-12-tetradecenyl acetate (E12-14:Ac) which are two sex pheromone compounds, and n-tetradecyl acetate (14:Ac) which has variable roles in mediating behavioral responses.  The ratios of these three compounds produced in female gland are geographically distinct among different populations.  Quantitative analysis of pheromone production showed that the proportions of Z12-14:Ac in the E/Z isomers (i.e., Z and E12-14:Ac) and the proportions of 14:Ac in the ternary blend respectively averaged 60.46% (SD=5.26) and 25.00% (SD=7.37), with their probabilities normally or near-normally distributed.  Trapping experiments in a cornfield indicated that deploying the E/Z isomers and the three compounds in rubber septa close to their gland ratios yielded the most captured males, while other ratios that deviated from the gland ratios showed reduced field captures.  The ternary blend was significantly more attractive to males than the E/Z isomers in the field, indicating a functional role of 14:Ac as the third pheromone component used by the local population.  Additionally, the dose-response test demonstrated that the application of the three compounds at dosages between 200 and 350 μg attracted significantly more males compared to other dosages.  Therefore, the characterization of this local ACB pheromone system provides additional information about its geographic variation and serves as a basis for optimizing the pheromone-mediated control of this pest in Xinjiang. 

Efficient and stable expression of foreign genes in cells and transgenic animals is important for gain-of-function studies and the establishment of bioreactors.  Safe harbor loci in the animal genome enable consistent overexpression of foreign genes, without side effects.  However, relatively few safe harbor loci are available in pigs, a fact which has impeded the development of multi-transgenic pig research.  We report a strategy for efficient transgene knock-in in the endogenous collagen type I alpha 1 chain (COL1A1) gene using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system.  After the knock-in of a 2A peptide-green fluorescence protein (2A-GFP) transgene in the last codon of COL1A1 in multiple porcine cells, including porcine kidney epithelial (PK15), porcine embryonic fibroblast (PEF) and porcine intestinal epithelial (IPI-2I) cells, quantitative PCR (qPCR), Western blotting, RNA-seq and CCK8 assay were performed to assess the safety of COL1A1 locus.  The qPCR results showed that the GFP knock-in had no effect (P=0.29, P=0.66 and P=0.20 for PK15, PEF and IPI-2I cells, respectively) on the mRNA expression of COL1A1 gene.  Similarly, no significant differences (P=0.64, P=0.48 and P=0.80 for PK15, PEF and IPI-2I cells, respectively) were found between the GFP knock-in and wild type cells by Western blotting.  RNA-seq results revealed that the transcriptome of GFP knock-in PEF cells had a significant positive correlation (P<2.2e–16) with that of the wild type cells, indicating that the GFP knock-in did not alter the global expression of endogenous genes.  Furthermore, the CCK8 assay showed that the GFP knock-in events had no adverse effects (P_24h=0.31, P_48h=0.96, P_72h=0.24, P_96h=0.17, and P_120h=0.38) on cell proliferation of PK15 cells.  These results indicate that the COL1A1 locus can be used as a safe harbor for foreign genes knock-in into the pig genome and can be broadly applied to farm animal breeding and biomedical model establishment

Soybean is one of the most important food crops worldwide.  Like other legumes, soybean can form symbiotic relationships with Rhizobium species.  Nitrogen fixation of soybean via its symbiosis with Rhizobium is pivotal for sustainable agriculture.  Type III effectors (T3Es) are essential regulators of the establishment of the symbiosis, and nodule number is a feature of nitrogen-affected nodulation.  However, genes encoding T3Es at quantitative trait loci (QTLs) related to nodulation have rarely been identified. Chromosome segment substitution lines (CSSLs) have a common genetic background but only a few loci with heterogeneous genetic information; thus, they are suitable materials for identifying candidate genes at a target locus.  In this study, a CSSL population was used to identify the QTLs related to nodule number in soybean.  Single nucleotide polymorphism (SNP) markers and candidate genes within the QTLs interval were detected, and it was determined which genes showed differential expression between isolines.  Four candidate genes (GmCDPK28, GmNAC1, GmbHLH, and GmERF5) linked to the SNPs were identified as being related to nodule traits and pivotal processes and pathways involved in symbiosis establishment.  A candidate gene (GmERF5) encoding a transcription factor that may interact directly with the T3E NopAA was identified.  The confirmed CSSLs with important segments and candidate genes identified in this study are valuable resources for further studies on the genetic network and T3Es involved in the signaling pathway that is essential for symbiosis establishment. 

The loss of N in farmland is an important cause of agricultural non-point source pollution, which seriously impacts the aquatic environment.  A two-year (2017–2018) experiment was conducted to investigate the characteristics of runoff and N losses under different tillage practices.  Taking downslope ridge planting and cross ridge planting as the experimental treatments, the characteristics of surface runoff, interflow, and N losses in sloping farmlands with yellow soil were studied throughout the maize growth period.  As the rainfall increased, the surface runoff and interflow also increased.  The surface runoff and N losses in the surface runoff of downslope ridge planting were significantly higher than those of cross ridge planting.  The interflow volumes and N losses in the 0–20 and 20–40 cm soil layers of the cross ridge planting were significantly higher than those of the downslope ridge planting.  The total N (TN) losses from surface runoff accounted for 54.95–81.25% of the N losses from all pathways.  Therefore, we inferred that surface runoff is the main pathway of N losses.  Dissolved total N (DTN) was the main form of N loss under different tillage measures, as it accounted for 55.82–94.41% of the TN losses, and dissolved organic N accounted for 52.81–87.06% of the DTN losses.  Thus, we inferred that dissolved N is the main form of N loss.  Future research must focus on the prevention and control of the N losses during the maize seedling stage to reduce the environmental pollution caused by ammonium N through runoff.

Starch is the most important component in endosperm of sorghum grain.  Usually, two types of starch are present: amylose (AM) and amylopectin (AP).  The levels of AM and AP contents play a significant role in the appearance, structure, and quality of sorghum grains and in marketing applications.  In the present study, a panel of 634 sorghum (Sorghum bicolor (L.) Moench) accessions were evaluated for starch, AM, and AP contents of grain, which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in India, and 252 landraces and 140 cultivars from China.  The average starch content was 67.64% and the average AM and AP contents were 20.19 and 79.81%, respectively.  We developed a total of 260 000 high-confidence single nucleotide polymorphism (SNP) markers in the panel of 634 accessions of S. bicolor using specific locus amplified fragment sequencing (SLAF-seq).  We performed genome-wide association studies (GWAS) of starch, AM, and AM/AP of grain and SNP markers based on a mixed linear model (MLM).  In total, 70 significant association signals were detected for starch, AM, and AM/AP ratio of grain with P<4.452×10^–7, of which 10 SNPs were identified with significant starch, 51 SNPs were associated with AM, and nine SNPs were associated with the AM/AP ratio.  The Gene Ontology (GO) analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals, located on chromosomes 1, 5, 6, and 9.  Of these genes, Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt (HMS) and Sobic.009G071800 encodes 6-phospho-fructokinase (PFK), which is involved in the embden-meyerhof pathway (EMP).  Kompetitive allele specific PCR (KASP) markers were developed to validate the GWAS results.  The C allele is correlated with a high starch content, while the T allele is linked with a low level of starch content, and provides reliable haplotypes for MAS in sorghum quality improvement.

Cotton plants are recalcitrant with regards to transformation and induced regeneration.  In the present study, 5-enolpyruvylshikimate-3-phosphate (EPSPS), a glyphosate resistant gene from the bacterium Agrobacterium sp. strain CP4, was introduced into an elite Bt transgenic cotton cultivar with a modified technique involving in planta Agrobacterium-mediated transformation of shoot apex.  Primary transformants were initially screened using a 0.26% glyphosate spray and subsequently by PCR analysis.  Five out of 4 000 transformants from T₁ seeds were obtained resulting in an in planta transformation rate of 0.125%.  Four homozygous lines were produced by continuous self-fertilization and both PCR-based selection and glyphosate resistance.  Transgene insertion was analyzed by Southern blot analysis.  Gene transcription and protein expression levels in the transgenic cotton lines were further investigated by RT-PCR, Western blot, and ELISA methods.  Transgenic T₃ plants were resistant to as much as 0.4% of glyphosate treatments in field trials.  Our results indicate that the cotton shoot apex transformation technique which is both tissue-culture and genotype-independent would enable the exploitation of transgene technology in different cotton cultivars.  Since this method does not require sterile conditions, the use of specialized growth media or the application of plant hormones, it can be conducted under the greenhouse condition.

Radiation sensitivity proteins-23 (RAD23) are DNA repair factors participate in the ubiquitin/proteasome system (UPS).  Although the genome-wide analysis of RAD23 family members has been conducted in some species, little is known about RAD23 genes in apple (Malus×domestica Borkh.).  We analyzed this gene family in M. domestica in terms of genomic locations, protein and promoter structures, and expressions in response to stresses.  Various members showed a ubiquitous pattern of expression in all selected apple parts.  Their expressions were altered under chilling, heat, and hydrogen peroxide treatments, as well as abscisic acid (ABA) treatment and water deficiency, suggesting their possible roles in plant stress responses.  These results provide essential information about RAD23 genes in apple and will contribute to further functional studies

Zinc (Zn) deficiency is widespread among citrus plants, but information about the mechanisms for Zn deficiency response in these plants is scarce. In the present study, different navel orange (Citrus sinensis (L.) Osbeck) leaves with various yellowing levels were sampled in our experimental orchard, and upon estimation of nutrient contents, Zn deficiencies were diagnosed as mild, moderate, and severe. Further analysis of chlorophyll content, photosynthetic characteristics, antioxidant enzyme activities, and expression levels of Zn/Iron-regulated transporter-like protein (ZIP) family genes were conducted in the sampled Zn-deficient leaves. The results showed that chlorophyll contents and net photosynthetic rate (P_n) seemed to decrease with reduced Zn contents. In addition, comparison of severe Zn-deficient and normal leaves revealed that activities of peroxidase (POD) and catalase (CAT) increased significantly, whereas that of Zn-containing enzymes such as Cu/Zn superoxide dismutase (Cu/Zn-SOD) significantly reduced with decreasing Zn contents. As expected, expression of the ZIP family genes, ZIP1, ZIP3, and ZIP4, was induced by Zn deficiencies. These results deepen our understanding of Zn deficiency in citrus plants as well as provide useful preliminary information for further research.

China is in a dominant position in apple production globally with both the largest apple growing area and the largest export of fresh apple fruits. However, the annual productivity of China’s apple is significantly lower than that of other dominant apple producing countries. In addition, apple production is based on excessive application of chemical fertilizers and the nutrient use efficiency (especially nitrogen) is therefore low and the nutrient emissions to the environment are high. Apple production in China is considerably contributes to farmers’ incomes and is important as export product. There is an urgent need to enhance apple productivity and improve nutrient use efficiencies in intensive apple production systems in the country. These can be attained by improved understanding of production potential, yield gaps, nutrient use and best management in apple orchards. To the end, priorities in research on apple production systems and required political support are described which may lead to more sustainable and environmental-friendly intensification of apple production in China.

    The vacuolar proton pump ATPase (V-H⁺-ATPase), which is a multi-subunit membrane protein complex, plays a major role in the activation of ion and nutrient transport and has been suggested to be involved in several physiological processes, such as cell expansion and salt tolerance. In this study, three genes encoding V-H⁺-ATPase subunits B (ScVHA-B, GenBank: JF826506), C (ScVHA-C, GenBank: JF826507) and H (ScVHA-H, GenBank: JF826508) were isolated from the halophyte Suaeda corniculata. The transcript levels of ScVHA-B, ScVHA-C and ScVHA-H were increased by salt, drought and saline-alkali treatments. V-H⁺-ATPase activity was also examined under salt, drought and saline-alkali stresses. The results showed that V-H⁺-ATPase activity was correlated with salt, drought and saline-alkali stress. Furthermore, V-H⁺-ATPase subunits B, C and H (ScVHA-B, ScVHA-C and ScVHA-H) from S. corniculata were introduced separately into the alfalfa genome. The transgenic alfalfa was verified by Southern and Northern blot analysis. During salt and saline-alkali stresses, transgenic linevacuolar proton pump, salt tolerance, saline-alkali tolerance, alfalfa
s carrying the B, C and H subunits had higher germination rates than the wild type (WT). More free proline, higher superoxide dismutase (SOD) activity and lower malondialdehyde (MDA) levels were detected in the transgenic plants under salt and saline-alkali treatments. Moreover, the ScVHA-B transgenic lines showed greater tolerance to salt and saline-alkali stresses than the WT. These results suggest that overexpression of ScVHA-B, ScVHA-C and ScVHA-H improves tolerance to salt and saline-alkali stresses in transgenic alfalfa.

Aeromonas hydrophila isolates from clinical cases (n=43) were tested against 8 antimicrobial agents and typed by outer membrane protein (OMP) pattern by using sodium dodecyl sulfate gel electrophoresis. All isolates were resistant to ampicillin (MICs, 16 mg mL-1) and sulfamonomethoxine (MICs, 64 mg mL-1), but susceptible to norfloxacin (MICs, 0.5 mg mL-1). There was a high incidence of resistance to erythromycin (90.70%) and tylosin (93.02%), while a low incidences of resistance to ciprofloxacin (2.33%), enrofloxacin (2.33%) and florfenicol (4.65%). Six different outer membrane protein patterns were found among 34 isolates by analyzing proteins in the range of 22 to 50 kDa, other than 9 isolates with their respective profiles. The strains with the similar OMP profiles had similar resistances. Compared with the other strains from the same OMP patterns, NB-1, A.Pun and MR-1 had lacked the proteins in the range of 30 to 45 kDa and their resistance to florfenicol substantially increased. It is speculated that the outer membrane protein changes might correlate with decreased susceptibility to florfenicol in the three strains. Some strains which showed completely identical OMP types had a little difference in their resistance to fluoroquinolones, indicating that there might be other factors that were involved in the antimicrobial resistance of A. hydrophila.