Rapeseed (Brassica napus L.) harvesting method is critical since it significantly determines the seed yield, oil quality, and industrial efficiency.  This study investigated the influences of harvesting methods on the quality of cold-pressed rapeseed oil of two varieties.  Oil color, peroxide value (POV), tocopherol content, fatty acid composition, and polarity of total polyphenols (PTP) contents of two rapeseed varieties in Huanggang and Xiangyang were compared through artificially simulated combined harvesting and two-stage harvesting.  Results showed significant differences in the quality of rapeseed oil between the two harvesting methods.  The red value (R-value), POV, total tocopherol contents, linoleic and linolenic acid content, and PTP content of the pressed rapeseed oil prepared by the combined harvesting method were about 27.6, 5.7, 15.8, 2.0, 0.5, and 28.6% lower than those of the oil produced from the two-stage harvesting method, respectively.  Xiangyang and Huayouza62 performed better in the two regions and two varieties, respectively.  To sum up, the rapeseed oil obtained 41–44 days after final flowering of combined harvesting, 35 days after final flowering, and six days of post-ripening of the two-stage harvesting had the best quality.

Herbivore-induced plant volatiles (HIPVs) play a key role in the interactions between plants and herbivorous insects, as HIPVs can promote or deter herbivorous insects’ behavior.  While aphids are common and serious phloem-feeding pests in farmland ecosystems, little is known about how aphids use their sensitive olfactory system to detect HIPVs.  In this study, the antennal transcriptomes of the aphid species Megoura crassicauda were sequenced, and expression level analyses of M. crassicauda odorant receptors (ORs) were carried out.  To investigate the chemoreception mechanisms that M. crassicauda uses to detect HIPVs, we performed in vitro functional studies of the ORs using 11 HIPVs reported to be released by aphid-infested plants.  In total, 54 candidate chemosensory genes were identified, among which 20 genes were ORs.  McraOR20 and McraOR43 were selected for further functional characterization because their homologs in aphids were quite conserved and their expression levels in antennae of M. crassicauda were relatively high.  The results showed that McraOR20 specifically detected cis-jasmone, as did its ortholog ApisOR20 from the pea aphid Acyrthosiphon pisum, while McraOR43 did not respond to any of the HIPV chemicals that were tested.  This study characterized the ability of the homologous OR20 receptors in the two aphid species to detect HIPV cis-jasmone, and provides a candidate olfactory target for mediating aphid behaviors.  

Soil organic carbon (SOC) and nitrogen (N) are two of the most important indicators for agricultural productivity.  The primary objective of this study was to investigate the changes in SOC and N in the deep soil profile (up to 100 cm) and their relationships with crop productivity under the influence of long-term (since 1990) fertilization  in the wheat-maize cropping system.  Treatments included CK (control), NP (inorganic N and phosphorus (P) fertilizers), NPK (inorganic N, P and potassium fertilizers), NPKM (NPK plus manure), and M (manure).  Crop yield and the properties of topsoil were measured yearly from 2001 to 2009.  C and N contents were measured at five different depths in 2001 and 2009.  The results showed that wheat and maize yields decreased between 2001 and 2009 under the inorganic fertilizer (NP and NPK) treatments.  The average yield between 2001 and 2009 under the NP, NPK, NPKM, and M treatments (compared with the CK treatment) increased by 38, 115, 383, and 381%, respectively, for wheat and 348, 891, 2 738, and 1 845%, respectively, for maize.  Different long-term fertilization treatments significantly changed coarse free particulate (cfPOC), fine free particulate (ffPOC), intramicroaggregate particulate (iPOC), and mineral-associated (mSOC) organic carbon fractions.  In the experimental years of 2001 and 2009, soil fractions occurred in the following order for all treatments: mSOC>cfPOC>iPOC>ffPOC.  All fractions were higher under the manure application treatments than under the inorganic fertilization treatments.  Compared to the inorganic fertilization treatments, manure input enhanced the stocks of SOC and total N in the surface layer (0–20 cm) but decreased SOC and N in the deep soil layer (80–100 cm).  This reveals the efficiency of manure in increasing yield productivity and decreasing risk of vertical loss of nutrients, especially N, compared to inorganic fertilization treatments.  The findings provide opportunities for understanding deep soil C and N dynamics, which could help mitigate climate change impact on agricultural production and maintain soil health.

Nitrogen (N) loss from fertilization in agricultural fields has an unavoidable negative impact on the environment and a better
understanding of the major pathways can assist in developing the best management practices. The aim of this study was
to evaluate the fate of N fertilizers applied to acidic red soil (Ferralic Cambisol) after 19 years of mineral (synthetic) and
manure fertilizer treatments under a cropping system with wheat-maize rotations. Five field treatments were examined:
control (CK), chemical nitrogen and potash fertilizer (NK), chemical nitrogen and phosphorus fertilizer (NP), chemical nitrogen,
phosphorus and potash fertilizer (NPK) and the NPK with manure (NPKM, 70% N from manure). Based on the soil
total N storage change in 0–100 cm depth, ammonia (NH3) volatilization, nitrous oxide (N₂O) emission, N plant uptake, and
the potential N leaching loss were estimated using a mass balance approach. In contrast to the NPKM, all mineral fertilizer
treatments (NK, NP and NPK) showed increased nitrate (NO₃^–-N) concentration with increasing soil depth, indicating higher
leaching potential. However, total NH3 volatilization loss was much higher in the NPKM (19.7%) than other mineral fertilizer
treatments (≤4.2%). The N₂O emissions were generally low (0.2–0.9%, the highest from the NPKM). Total gaseous loss
accounted for 1.7, 3.3, 5.1, and 21.9% for NK, NP, NPK, and NPKM treatments, respectively. Estimated N leaching loss
from the NPKM was only about 5% of the losses from mineral fertilizer treatments. All data demonstrated that manure
incorporation improved soil productivity, increased yield, and reduced potential leaching, but with significantly higher NH₃
volatilization, which could be reduced by improving the application method. This study confirms that manure incorporation is an essential strategy in N fertilization management in upland red soil cropping system.

Sucrose is known to play an important role in the cryopreservation of sperm and female gonads; however, its effect on the cryopreservation of pig spermatogonial stem cells (pSSCs) has not been tested.  The aim of this work was to study the effect of sucrose during pSSC cryopreservation and to find the most effective concentration in freezing medium.  pSSCs were cryopreserved with freezing media containing different concentrations of sucrose (70, 140, 210, and 280 mmol L^–1) and a control group without sucrose.  The survival rates, plasma membrane integrity, and mitochondrial membrane potential of thawed cells were detected by trypan blue (TB) staining, SYBR-14/propidium iodide (PI) dual staining, and JC-1 staining, respectively.  All the staining results showed an obvious increase in cell survival in the sucrose-treated groups as compared to that in the control group, with the exception of 280 mmol L^–1 sucrose.  Moreover, the 210 mmol L^–1 sucrose group yielded the highest survival rate among all the groups (P<0.05).  The results of SYBR-14/PI dual staining and JC-1 staining were consistent with those of TB staining as above described.  Quantitative real-time PCR (qRT-PCR) indicated that the mRNA levels of three apoptosis-promoting genes (BAX, APAF1 and CASPASE9) were significantly higher in thawed cells than in cells before freezing (P<0.05).  Moreover, the mRNA level of one anti-apoptotic gene (XIAP) was significantly lower in thawed cells than in cells before freezing (P<0.05).  When comparing the mRNA expression of apoptosis-related genes in thawed cells, the mRNA level of the anti-apoptotic genes in the control group was significantly lower than that in the sucrose-treated
groups (P<0.05).  Western blot analyses showed that the expression levels of cleaved CASPASE9, CASPASE3 and PARP-1 in the sucrose-treated groups were lower than those in the control group and were the lowest in the 210 mmol L^–1 sucrose group.  Both qRT-PCR and Western blot analyses suggested that sucrose inhibited cell apoptosis during freezing and thawing.  Briefly, sucrose promoted pSSCs survival after freezing and thawing, especially at a concentration of 210 mmol L^–1, which possibly assisted pSSC dehydration and inhibited cell apoptosis.  These findings hold great promise for further studies of the regulatory mechanism of proliferation and differentiation of pSSCs. 

To decipher the relationship between the inhibited shoot growth and expression pattern of key enzymes in nitrogen metabolism under root restriction, the effects of root restriction on diurnal variation of expression of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS1-1, GS1-2, GS2) and glutamate synthase (Fd-GOGAT, NADH-GOGAT) genes and nitrogen levels were evaluated in two-year-old Jumeigui grapevines (Vitis vinifera L.×Vitis labrusca L.) when significant differences in shoot growth were observed between treatments at expansion stage (22 days after anthesis). Grapevines were planted in root-restricting pits as root restriction and in an unrestricted field as the control. Results showed that root restriction significantly reduced shoot growth, but promoted the growth of white roots and fibrous brown roots and improved the fruit quality. (NO3 –+NO2 –)-N concentration in all plant parts, NH4 +-N concentration in white roots and total N concentration in leaves and brown roots were significantly reduced under root restriction. Gene expression analysis revealed that mRNA levels of genes related to the GS1/NADH-GOGAT pathway were lower in root-restricted than in control petioles, whereas genes involved in the GS2/Fd-GOGAT pathway were up-regulated under root restriction. Root restriction also resulted in downregulation of genes involved in nitrogen metabolism in leaves, especially at 10:00, while transcript levels of all these genes were enhanced in root-restricted white and brown roots at most time points. This organ-dependent response contributed to the alteration in NO3 – reduction and NH4 + assimilation under root restriction, leading to less NO3 – transported from roots and then assimilated in root-restricted leaves. Therefore, this study implied that shoot growth inhibition in grapevines under root restriction is closely associated with down-regulation of gene expression in nitrogen metabolism in leaves.

Upland red soils have been identified as major CO2 and N2O sources induced by human activities such as fertilization. To monitor characteristics of soil surface CO2 and N2O fluxes in cropland ecosystems after continuous fertilizer applications over decades and to separate the respective contributions of root and heterotrophic respiration to the total soil CO2 and N2O fluxes, the measurements of soil surface CO2 and N2O fluxes throughout the maize growing season in 2009 were carried out based on a fertilization experiment (from 1990) through of the maize (Zea mays L.) growing season in red soil in southern China. Five fertilization treatments were chosen from the experiment for study: zero-fertilizer application (CK), nitrogen-phosphorus- potassium (NPK) fertilizer application only, pig manure (M), NPK plus pig manure (NPKM) and NPK with straw (NPKS). Six chambers were installed in each plot. Three of them are in the inter-row soil (NR) and the others are in the soil within the row (R). Each fertilizer treatment received the same amount of N (300 kg ha-1 yr-1). Results showed that cumulative soil CO2 fluxes in NR or R were both following the order: NPKS>M, NPKM>NPK>CK. The contributions of root respiration to soil CO2 fluxes was 40, 44, 50, 47 and 35% in CK, NPK, NPKM, M and NPKS treatments, respectively, with the mean value of 43%. Cumulative soil N2O fluxes in NR or R were both following the order: NPKS, NPKM>M>NPK>CK, and soil N2O fluxes in R were 18, 20 and 30% higher than that in NR in NPKM, M and NPKS treatments, respectively, but with no difference between NR and R in NPK treatment. Furthermore, combine with soil temperature at -5 cm depth and soil moisture (0-20 cm) together could explain 55-70% and 42-59% of soil CO2 and N2O emissions with root interference and 62- 78% and 44-63% of that without root interference, respectively. In addition, soil CO2 and N2O fluxes per unit yield in NPKM (0.55 and 0.10 kg C t-1) and M (0.65 and 0.13 g N t-1) treatments were lower than those in other treatments. Therefore, manure application could be a preferred fertilization strategy in red soils in South China.

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.

A long-term field experiment was established to determine the influence of mineral fertilizer and organic manure on soil fertility. A tract of red soil (Ferralic Cambisol) in Qiyang Red Soil Experimental Station (Qiyang County, Hunan Province, China) was fertilized beginning in 1990 and N2O and CO2 were examined during the maize and wheat growth season of 2007-2008. The study involved five treatments: organic manure (NPKM), fertilizer NPK (NPK), fertilizer NP (NP), fertilizer NK (NK), and control (CK). Manured soils had higher crop biomass, organic C, and pH than soils receiving the various mineralized fertilizers indicating that long-term application of manures could efficiently prevent red soil acidification and increase crop productivity. The application of manures and fertilizers at a rate of 300 kg N ha-1 yr-1 obviously increased N2O and CO2 emissions from 0.58 kg N2O-N ha-1 yr-1 and 10 565 kg C ha-1 yr-1 in the CK treatment soil to 3.01 kg N2O-N ha-1 yr-1 and 28 663 kg C ha-1 yr-1 in the NPKM treatment. There were also obvious different effects on N2O and CO2 emissions between applying fertilizer and manure. More N2O and CO2 released during the 184-d maize growing season than the 125- d wheat growth season in the manure fertilized soils but not in mineral fertilizer treatments. N2O emission was significantly affected by soil moisture only during the wheat growing season, and CO2 emission was affected by soil temperature only in CK and NP treatment during the wheat and maize growing season. In sum, this study indicates the application of organic manure may be a preferred strategy for maintaining red soil productivity, but may result in greater N2O and CO2 emissions than treatments only with mineral fertilizer.