Elevating soil water content (SWC) through irrigation was one of the simple mitigation measures to improve crop resilience to heat stress.  The response of leaf function, such as photosynthetic capacity based on chlorophyll fluorescence during the mitigation, has received limited attention, especially in field conditions.  A two-year field experiment with three treatments (control treatment (CK), high-temperature treatment (H), and high-temperature together with elevating SWC treatment (HW)) was carried out during grain filling with two maize hybrids at a typical station in North China Plain.  Averagely, the net photosynthetic rate (P_n) was improved by 20%, and the canopy temperature decreased by 1–3°C in HW compared with in H in both years.  Furthermore, the higher SWC in HW significantly improved the actual photosynthetic rate (Phi2), linear electron flow (LEF), variable fluorescence (F_v), and the maximal potential quantum efficiency (F_v/F_m) for both hybrids.  Meanwhile, different responses in chlorophyll fluorescence between hybrids were also observed.  The higher SWC in HW significantly improved thylakoid proton conductivity (gH⁺) and the maximal fluorescence (F_m) for the hybrid ZD958.  For the hybrid XY335, the proton conductivity of chloroplast ATP synthase (vH⁺) and the minimal fluorescence (F_o) was increased by the SWC.  The structural equation model (SEM) further showed that SWC had significantly positive relationships with P_n, LEF, and F_v/F_m.  The elevating SWC alleviated heat stress with the delayed leaf senescence to prolong the effective period of photosynthesis and enhanced leaf photosynthetic capacity by improving Phi2, LEF, F_v, and F_v/F_m.  This research demonstrates that elevating SWC through enhancing leaf photosynthesis during grain filling would be an important mitigation strategy for adapting to the warming climate in maize production.

African swine fever (ASF), caused by the African swine fever virus (ASFV), is an acute, hemorrhagic, and contagious disease of domestic pigs and wild boars.  The disease is notifiable and listed by the World Organization for Animal Health (WOAH) (Wang N et al. 2019).  The outcomes of ASF infection can be peracute, acute, subacute, and chronic, depending on the virulence of ASFVs.  According to the report of WOAH (https://www.woah.org/app/uploads/2022/12/asf-report24.pdf), from January 2020 to December 2022, ASF led to more than  2 million pig losses.  Currently, ASFV persists continuously in more than 23 countries and poses a serious threat to the global swine industry.  ASF invaded China on 3 August, 2018, caused by genotype II virulent Georgia-07-like ASFVs (Wen et al. 2019; Zhao et al. 2019; Wang et al. 2020; Wang L et al. 2022).  An experimental study showed that Georgia-07-like ASFV HLJ/18 isolated in China is highly lethal and efficiently transmissible in domestic pigs (Zhao et al. 2019; Jiang et al. 2021).  During the past four years, genotype II Georgia-07-like ASFVs dominantly spread in China.  However, the low virulent genotype II and I ASFVs have been successively reported in China in 2020 and 2021, respectively (Sun et al. 2021a, b; Shi et al. 2022).  Compared with the high virulent genotype II HLJ/18 strain, the low virulent genotype I and II ASFVs had lower virulence and high transmissibility in pigs and induced persistent and chronic infection showing irregular virus shedding at low levels (Sun et al. 2021a, b; Tsegay et al. 2022; Wang P et al. 2022).  Notably, when different genotype I and genotype II viruses infect the same pig in the field, a novel virus may be generated through viral genome recombination, which brings new problems and challenges for the prevention and control of ASF in China.  Thus, a diagnostic method that differentiates genotype I and II ASFVs with high sensitivity and stability is urgently needed and will be helpful for the prevention and control of ASF in China.  

ASFVs have been divided into at least 24 genotypes based on the C-terminus of the B646L gene with 478 nt (Bastos et al. 2003).  B646L gene is one of the most used target genes for ASF diagnosis, which is also the target gene for the WOAH recommended PCR and fluorescent quantitative PCR assays (Agüero et al. 2003; King et al. 2003).  Sanger sequencing of targeted amplification of the B646L genes is the main genotyping approach for ASFVs.  Recently, Li et al. (2022) developed the duplex real-time PCR assay based on the ASFV E296R gene, and Cao et al. (2022) established the TaqMAN-MGB probe assay based on the N-terminal sequences of the B646L gene (Cao et al. 2022; Li et al. 2022), which could distinguish genotype I and II ASFVs with detection limits of 10 copies.  However, the target genes or regions in their methods were out of ASFV genotyping regions.  

Single nucleotide polymorphism (SNP) is a single base change at a specific position in the genome of different individuals and can be used as a genotyping marker for the detection of different individual genotypes (Gut 2001).  The amplification refractory mutation system (ARMS), also named Allele-specific PCR (AS-PCR), relies on the extension of primer only when its 3´ end has a perfect complement to the template (Wang M et al. 2019).  ARMS-qPCR technology has been developed and widely used in SNP detection and genotyping (Ochsenreither et al. 2010; Shi et al. 2013; Wang M et al. 2019).  Compared with other assays for SNP detection and genotyping, ARMS-qPCR has the advantage of low-cost, simple operation, high sensitivity, and rapid and real-time detection.

Here, 126 complete or partial B646L genes of ASFVs, including 78 genotype I and 48 genotype II viruses, were obtained from the GenBank database, and their information is shown in Appendix A.  After analyzing these genes by the MegAlign Software (DNAStar), there were 4 SNPs in the C-terminus of the B646L gene, differentiating genotype I viruses from genotype II viruses (Fig. 1-A).  Two SNPs at sites 1 656 and 1 710 were used to design primers and probes for differential detection of genotype I and II ASFVs (Fig. 1-A).  As previously described (Huang et al. 1992; Liu et al. 2012), primers (I F, II F and R) and probes (probe 1 and probe 2) were designed with the targeted gene sequences using Primer 5 Software (Fig. 1-B; Appendix B).  The duplex ARMS-qPCR reaction system volume was 25 μL: 12.5 μL of 2× HyperProbe Mixture (GENFINE), 0.5 μL of I F, II F and R primers (10 μmol L^–1), 0.5 μL of probe 1 and probe 2 (10 μmol L^–1), 5 μL of template DNA, and 5 μL of ddH₂O.  The duplex ARMS-qPCR was performed by using the Bio-Rad CFX96 Touch Real-Time PCR Detection System with the following reaction conditions: 95°C for 30 s, followed by 40 cycles of 95°C for 10 s, and 60°C for 30 s.  Fluorescence signal was detected at the end of each cycle of extension step.  For the positive sample of genotype I ASFV, FAM and Cy5 fluorophores could be detected; however, for the positive sample of genotype II ASFV, only FAM fluorophore could be detected (Fig. 1-B).  

The standard curve test revealed that for the standard plasmids of genotype I ASFV, the slopes were –3.3825 for Cy5 and –3.1906 for FAM; the correlation coefficient R² was 0.999 for Cy5 and 0.998 for FAM; the amplification efficiency was 97.53% for Cy5 and 100.06% for FAM, respectively (Fig. 1-C); for the standard plasmids of genotype II ASFV, the slope was –3.2983 for FAM, the correlation coefficient R² was 0.992 for FAM, the amplification efficiency was 100.01% for FAM, whereas Cy5 fluorophore could not be detected (Fig. 1-C).  In addition, the sensitivity of the duplex ARMS-qPCR was 10 copies per reaction for both genotype I and II ASFVs (Fig. 1-D).  Thus, these results indicated that the duplex ARMS-qPCR assay has high efficiency and sensitivity.  

We then evaluated the specificity of the duplex ARMS-qPCR.  The nucleic acids of 7 other swine viruses, including PRRSV, CSFV, PRV, PCV2, PEDV, TGEV, and PoRV, were used as templates.  There were 3 amplification curves obtained for genotype I ASFV (FAM and Cy5 signals) and II ASFV (FAM signal), whereas no amplification curve was recorded for the nucleic acids of PRRSV, CSFV, PRV, PCV2, PEDV, TGEV, and PoRV, as well as genotype II ASFV (Cy5 signal) and ddH₂O (Fig. 1-E).  The results demonstrated that the duplex ARMS-qPCR has a good specificity without cross-reactivity with other swine viruses.

The results of the stable detection limit test showed that for the standard plasmids of genotype I ASFV, all 12 replicates were tested positive at the dilution of 10 copies, while 7/12 replicates were tested positive at the dilution of 5 copies (Fig. 1-F); for the standard plasmids of genotype II ASFV, all 12 replicates were tested positive at the dilution of 10 copies, while 6/12 replicates were tested positive at the dilution of 1 copy (Fig. 1-F).  Thus, the stable detection limit of the duplex ARMS-qPCR was 10 copies per reaction for both genotype I and II ASFVs (Fig. 1-F).

We further assessed the repeatability and reproducibility of the duplex ARMS-qPCR.  The assay tested the standard plasmids of 3 concentrations (10⁶, 10⁴, and 10² copies).  For the standard plasmids of genotype I ASFV, the intra- and inter-assay variation of Ct value for the duplex ARMS-qPCR ranged from 0.07 to 0.93% and 1.2 to 2.17% in FAM fluorescence channel and from 0.38 to 1.02% and 0.85 to 1.27% in Cy5 fluorescence channel, respectively (Table 1).  For the standard plasmids of genotype II ASFV, the intra- and inter-assay variation of Ct value for the duplex ARMS-qPCR ranged from 0.27 to 0.61% and 0.77 to 1.07% in FAM fluorescence channel (Table 1).  These findings suggested that the duplex ARMS-qPCR assay has satisfactory repeatability and reproducibility.

Finally, we evaluated the duplex ARMS-qPCR compared with WOAH-qPCR.  A total of 40 samples were detected using both assays, including blood, oral and rectal swabs, tissues, and cell cultures from pigs or PAMs infected by genotype I and II ASFVs.  Animal studies have evaluated the virulence and transmissibility of genotype I ASFV SD/DY-I/21 and genotype II virus HLJ/18 (Zhao et al. 2019; Sun et al. 2021a), respectively.  The results showed that 36 samples, including 18 of genotype I ASFV and 18 of genotype II ASFV were detected to be positive and differentiated by the duplex ARMS-qPCR, which were consistent with the results of the WOAH-qPCR (Appendix C).  

In summary, we developed a duplex ARMS-qPCR assay based on ASFV genotyping region of B646L gene, which can effectively differentiate genotype I and II ASFVs.  The assay had high sensitivity and specificity and exhibited good results in detecting samples, including blood, oral and rectal swabs, tissues, and cell culture.  Whether our method could be used for differentiating other genotypes of ASFVs is needed for further evalution.  However, just genotype I and II ASFVs are spreading outside Africa.  Thus, our method will provide an additional epidemiological investigation tool to implement effective ASFV control and prevention.

The present study modified potato protein and flour with tyrosinase to promote the diversification of potato staple foods.  The results indicated that tyrosinase treatment markedly altered the secondary structure of proteins.  After tyrosinase treatment, the maximum decomposition temperature of potato protein and flour increased from 322.32 to 332.40°C and from 294.24 to 299.61°C, respectively.  Tyrosinase treatment remarkably reduced the pasting viscosity of potato flour, that is, the peak viscosity, through reducing viscosity, breakdown, final viscosity, and setback by 32.50, 60.98, 13.04, 68.24, and 74.31%, respectively.  In contrast, tyrosinase treatment increased the shear resistance and hardness of the protein and flour gels; the maximum stress values of the protein and flour gels increased from 1.48 to 10.1% and from 6.87 to 14.8%, respectively.  Furthermore, tyrosinase treatment promoted viscoelastic properties and structural stability of potato protein and flour.  These results may provide an important foundation for the development of novel potato staple foods.

Investigations into the potential application of nanoparticles acting as nanofungicides in sustainable agriculture are rapidly expanding due to the high antimicrobial properties of these compounds, which do not risk inducing pathogen resistance to fungicides.  A detailed understanding of the impact of copper oxide nanoparticles (CuO NPs) on soil-borne phytopathogenic fungi is yet to be obtained.  This study aimed to explore the in vitro antifungal activity and control efficacy of CuO NPs applied via irrigation with respect to tobacco black shank (TBS) disease caused by Phytophthora nicotianae.  The results revealed that CuO NPs greatly interfered with the reproductive growth process of this fungus, repressing hyphal growth, spore germination and sporangium production.  Additionally, morphological damage, intracellular ROS accumulation and increased SOD enzyme activity in hyphae were the antifungicidal mechanisms of these NPs.  In pot experiments, treatment with CuO NPs at 100 mg L^–1 significantly suppressed TBS development, compared with the effect on control plants, and the control efficacy reached 33.69% without inducing phytotoxicity.  Exposure to CuO NPs significantly activated a series of defense enzymes, and resistance genes in tobacco can further explain the mechanisms by which CuO NPs suppressed fungal infection.  The Cu content in both the leaves and roots of P. nicotianae-infested plants increased by 50.03 and 27.25%, respectively, after treatment with 100 mg L^–1 CuO NPs, compared with that of healthy plants.  In particular, a higher Cu content was observed in infected roots than in leaves.  Therefore, this study showed the potential of CuO NPs applied as nanofungicides and as nanoinducers of fungus resistance genes for the management of TBS through inhibition of pathogen infection and stimulation of plant defenses.

Bovine parainfluenza virus type 3 (BPIV3) is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world.  BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C (BPIV3c).  Pathogen investigations had shown that BPIV3c infection was very common among cattle in China.  To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis.  Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays.  In the present study, the BPIV3c strain SD0835 was continuously passaged on Madin-Darby bovine kidney (MDBK) cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs.  The passage 209 of BPIV3c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs.  The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes.  The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3c strain SD0835.  Additionally, the percentages of CD4⁺ and CD8⁺ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4⁺ and CD8⁺ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group.  These data support further testing of the attenuated virus as an effective candidate vaccine.

Citrus yellow vein clearing virus (CYVCV) is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae.  Capsid protein (CP) of CYVCV Chongqing isolate (CYVCV-CQ) was produced using a prokaryotic expression system and used as the immunogen for monoclonal antibody (MAb) production.  Four highly specific and sensitive murine MAbs and one polyclonal antibody were prepared in this study.  Titers of the four MAbs in ascites fluids ranged from 10^–6 to 10^–7 as determined by indirect enzyme-linked immunosorbent assay (ELISA).  Three serological assays, including dot enzyme-linked immunosorbent assay (dot-ELISA), tissue blot-ELISA, and double-antibody sandwich (DAS)-ELISA, were developed for quick and reliable detections of CYVCV in citrus samples.  The developed dot-ELISA and DAS-ELISA methods could detect CYVCV in the infected citrus leaf crude extracts diluted at 1:2 560 and 1:10 240 (w/v, g mL^–1), respectively.  The detection result of 125 citrus leaf samples collected from citrus groves in Yunnan Province and Chongqing Municipality of China showed that approximately 36% samples were positive for CYVCV.  This virus was, however, not detected in any sample collected from Zhejiang or Jiangxi Province, China.

Water shortage has threatened sustainable development of agriculture globally as well as in the North China Plain (NCP).  Irrigation, as the most effective way to increase food production in dry land, may not be readily available in the situation of drought.  One of the alternatives is to supply plants with enough nutrients so that they can be more sustainable to the water stress.  The objective of this study was to explore effects of irrigation and sulphur (S) application on water consumption, dry matter accumulation (DMA), and grain yield of winter wheat in NCP.  Three irrigation regimes including no irrigation (rainfed, I₀) during the whole growth period, once irrigation only at jointing stage (90 mm, I₁), and twice respective irrigation at jointing and anthesis stages (90 mm plus 90 mm, I₂), and two levels of S application including 0 (S₀) and 60 kg ha^–1 (S₆₀) were designed in the field experiment in NCP.  Results showed that increasing irrigation times significantly increased mean grain yield of wheat by 12.5–23.7% and nitrogen partial factor productivity (NPFP) by 21.2–45.0% in two wheat seasons, but markedly decreased crop water use efficiency (YWUE).  Furthermore, S supply 60 kg ha^–1 significantly increased mean grain yield, YWUE, IWUE and NPFP by 5.6, 6.1, 23.2, and 5.6% (across two wheat seasons), respectively.  However, we also found that role of soil moisture prior to S application was one of important greater factors on improving the absorption and utilization of storage water and nutrients of soil.  Thus, water supply is still the most important factor to restrict the growth of wheat in the present case of NCP, supplying 60 kg ha^–1 S with once irrigation 90 mm at the jointing stage is a relatively appropriate recommended combination to improve grain yield and WUE of wheat when saving water resources is be considered in irrigated wheat farmlands of NCP.

Grain size is a major determinant of grain weight and a trait having important impact on grain quality in rice. The objective of this study is to detect QTLs for grain size in rice and identify important QTLs that have not been well characterized before. The QTL mapping was first performed using three recombinant inbred line populations derived from indica rice crosses Teqing/IRBB lines, Zhenshan 97/Milyang 46, Xieqingzao/Milyang 46. Fourteen QTLs for grain length and 10 QTLs for grain width were detected, including seven shared by two populations and 17 found in one population. Three of the seven common QTLs were found to coincide in position with those that have been cloned and the four others remained to be clarified. One of them, qGS10 located in the interval RM6100–RM228 on the long arm of chromosome 10, was validated using F2:3 populations and near isogenic lines derived from residual heterozygotes for the interval RM6100–RM228. The QTL was found to have a considerable effect on grain size and grain weight, and a small effect on grain number. This region was also previously detected for quality traits in rice in a number of studies, providing a good candidate for functional analysis and breeding utilization.

The current interest in the health benefits of whole wheat grain has prompted breeders to further increase the concentration of antioxidants in wheat. The objective of this study was to investigate the variation in antioxidant content among Chinese wheat grains and the relationship between antioxidants and grain color and morphological characteristics. A wide variation was observed in the total phenolic, carotenoid and flavonoid contents, as well as the antioxidant activity (AOA), of Chinese wheat varieties. Black wheat had the highest mean total phenolic, carotenoid and flavonoid contents and the highest AOA, followed by red and white wheats. The grain color parameters were significantly negatively correlated with total phenolic, carotenoid and flavonoid contents and AOA among all of the wheat varieties examined, and grain weight was also significantly negatively correlated with these traits. The same correlation between grain weight and antioxidant traits was also observed within individual groups of wheat, which indicates that grain weight may be used as an index for selecting wheat varieties with high AOA. Landraces had significantly higher flavonoid content than commercial wheat varieties. The results of this study may be useful for breeding nutrient-rich wheat varieties.

The extraction and comparison of soil amino acids using different extractants (deionized water, K2SO4, Na2SO4, NaCl, KCl) were reported. Results showed that 0.5 mol L-1 K2SO4 with a 5 times extraction was a better method to assess the concentration of extractable amino acids in soils. The total amino acids extracted from soil planted for tea were similar to the total inorganic nitrogen. While they extracted from vegetable soil and paddy soil were much lower than the total inorganic nitrogen.

Quantitative trait loci (QTL) for percentage of chalky grain, degree of chalkiness, and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of commercial three-line hybrids of indica rice. Two of the populations showed great variations on heading date, and the other had a short range of heading date variation. A total of 40 QTLs were detected and fell into 15 regions of 10 chromosomes, of which 5 regions were detected for 1 or more same traits over different populations, 2 were detected for different traits in different populations, 3 were detected for 2 or all the 3 traits in a single population, and 5 were detected for a single trait in a single population. Most of these QTLs have been reported previously, but a region located on the long arm of chromosome 10 showing significant effects in all the 3 populations has not been reported before. It was shown that a number of gene cloned, including the Wx and Alk for the physiochemical property of rice grain, and GW2, GS3 and GW5 for grain weight and grain size, could have played important roles for the genetic control of grain chalkiness in rice, but there are many more QTLs exerting stable effects for rice chalkiness over different genetic backgrounds. It is worth paying more attentions to these regions which harbor QTL such as the qPCG5.2/qDC5.2/qET5.2 and qPCG10/qDC10/qET10 detected in our study. Our results also showed that the use of segregating populations having high-uniform heading date could greatly increase the efficiency of the identification of QTL responsible for traits that are subjected to great environmental influence.