Reducing environmental impacts and improving N utilization are critical to ensuring food security in China.  Although root-zone fertilization has been considered an effective strategy to improve nitrogen use efficiency (NUE), the effect of controlled-release urea (CRU) applied in conjunction with normal urea in this mode is unclear.  Therefore, a 3-year field experiment was conducted using a no-N-added as a control and two fertilization modes (FF, furrow fertilization by manual trenching, i.e., farmer fertilizer practice; HF: root-zone hole fertilization by point broadcast manually) at 210 kg N ha^–1 (controlled-release:normal fertilizer=5:5), along with a 1-year in-situ microplot experiment.  Maize yield, NUE and N loss were investigated under different fertilization modes.  The results showed that compared with FF, HF improved the average yield and N recovery efficiency by 8.5 and 22.3% over three years, respectively.  HF had a greater potential for application than FF treatment, which led to increases in dry matter accumulation, total N uptake, SPAD value and LAI.  In addition, HF remarkably enhanced the accumulation of ¹⁵N derived from fertilizer by 17.2% compared with FF, which in turn reduced the potential loss of ¹⁵N by 43.8%.  HF increased the accumulation of N in the tillage layer of soils at harvest for potential use in the subsequent season relative to FF.  Hence, HF could match the N requirement of summer maize, sustain yield, improve NUE and reduce environmental N loss simultaneously.  Overall, root-zone hole fertilization with blended CRU and normal urea can represent an effective and promising practice to achieve environmental integrity and food security on the North China Plain, which deserves further application and investigation.

Photosynthesis occurs mainly in chloroplasts, whose development is regulated by proteins encoded by nuclear genes.  Among them, pentapeptide repeat (PPR) proteins participate in organelle RNA editing.  Although there are more than 450 members of the PPR protein family in rice, only a few affect RNA editing in rice chloroplasts.  Gene editing technology has created new rice germplasm and mutants, which could be used for rice breeding and gene function study.  This study evaluated the functions of OsPPR9 in chloroplast RNA editing in rice.  The osppr9 mutants were obtained by CRISPR/Cas9, which showed yellowing leaves and a lethal phenotype, with suppressed expression of genes associated with chloroplast development and accumulation of photosynthetic-related proteins.  In addition, loss of OsPPR9 protein function reduces the editing efficiency of rps8-C182, rpoC2-C4106, rps14-C80, and ndhB-C611 RNA editing sites, which affects chloroplast growth and development in rice.  Our data showed that OsPPR9 is highly expressed in rice leaves and encodes a DYW-PPR protein localized in chloroplasts.  Besides, the OsPPR9 protein was shown to interact with OsMORF2 and OsMORF9.  Together, our findings provide insights into the role of the PPR protein in regulating chloroplast development in rice. 

With the implementation of the C-strain vaccine, classical swine fever (CSF) has been under control in China, which is currently in a chronic atypical epidemic situation.  African swine fever (ASF) emerged in China in 2018 and spread quickly across the country. It is presently occurring sporadically due to the lack of commercial vaccines and farmers’ increased awareness of biosafety.  Atypical porcine pestivirus (APPV) was first detected in Guangdong Province, China, in 2016, which mainly harms piglets and has a local epidemic situation in southern China.  These three diseases have similar clinical symptoms in pig herds, which cause considerable losses to the pig industry.  They are difficult to be distinguished only by clinical diagnosis.  Therefore, developing an early and accurate simultaneous detection and differential diagnosis of the diseases induced by these viruses is essential.  In this study, three pairs of specific primers and Taq-man probes were designed from highly conserved genomic regions of CSFV (5´ UTR), African swine fever virus (ASFV) (B646L), and APPV (5´ UTR), followed by the optimization of reaction conditions to establish a multiplex real-time PCR detection assay.  The results showed that the method did not cross-react with other swine pathogens (porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV), porcine parvovirus (PPV), and bovine viral diarrhea virus BVDV).  The sensitivity results showed that CSFV, ASFV, and APPV could be detected as low as 1 copy mL^–1; the repeatability results showed that the intra-assay and inter-assay coefficient of variation of ASFV, CSFV, and APPV was less than 1%.  Twenty-two virus samples were detected by the multiplex real-time PCR, compared with national standard diagnostic and patented method assay for CSF (GB/T 27540–2011), ASF (GB/T 18648–2020), and APPV (CN108611442A), respectively.  The sensitivity of this triple real-time PCR for CSFV, ASFV, and APPV was almost the same, and the  compliance results were the same (100%).  A total of 451 clinical samples were detected, and the results showed that the positive rates of CSFV, ASFV, and APPV were 0.22% (1/451), 1.3% (6/451), and 0% (0/451), respectively.  This assay provides a valuale tool for rapid detection and accurate diagnosis of CSFV, ASFV, and APPV.

As important yield-related traits, thousand-grain weight (TGW), grain number per spike (GNS) and grain weight per spike (GWS) are crucial components of wheat production.  To dissect their underlying genetic basis, a double haploid (DH) population comprised of 198 lines derived from 8762/Keyi 5214 was constructed.  We then used genechip to genotype the DH population and integrated the yield-related traits TGW, GNS and GWS for QTL mapping.  Finally, we obtained a total of 18 942 polymorphic SNP markers and identified 41 crucial QTLs for these traits.  Three stable QTLs for TGW were identified on chromosomes 2D (QTgw-2D.3 and QTgw-2D.4) and 6A (QTgw-6A.1), with additive alleles all from the parent 8762, explaining 4.81–18.67% of the phenotypic variations.  Five stable QTLs for GNS on chromosomes 3D, 5B, 5D and 6A were identified.  QGns-5D.1 was from parent 8762, while the other four QTLs were from parent Keyi 5214, explaining 5.89–7.08% of the GNS phenotypic variations.  In addition, a stable GWS genetic locus QGws-4A.3 was detected from the parent 8762, which explained 6.08–6.14% of the phenotypic variations.  To utilize the identified QTLs, we developed STARP markers for four important QTLs, Tgw2D.3-2, Tgw2D.4-1, Tgw6A.1 and Gns3D.1.  Our results provide important basic resources and references for the identification and cloning of genes related to TGW, GNS and GWS in wheat.

Laccases, as a kind of multicopper oxidase, play an important role in pigment synthesis and growth in fungi and are involved in their interactions with host plants.  In Setosphaeria turcica, 9 laccase-like multicopper oxidases have been identified, and StLAC2 is involved in the synthesis of the melanin that accumulates in the cell wall.  The function of another major laccase gene, StLAC6, was studied here.  The knockout of StLAC6 had no effect on the growth, morphology or invasion ability of S. turcica, but the morphology and function of peroxisomes of knockout mutants were abnormal.  The knockout of the StLAC6 gene resulted in increased contents of phenolic compounds and melanin and the sensitivity to fungicides increased compared with wild type strains.  In the mutants of StLAC6, there is a significant change of the expression levels of other laccase genes.  This study provides a new insight into laccase functions and the relationship of the laccase gene family in plant pathogenic fungi.