The optimized management of crop fertilization is very important for improving crop yield and reducing the consumption of chemical fertilizers. Critical nutrient values can be used for evaluating the nutritional status of a crop, and they reflect the nutrient concentrations above which the plant is sufficiently supplied for achieving the maximum potential yield. Based on on-farm surveys of 504 farmers and 60 field experimental sites in the drylands of China, we proposed a recommended fertilization method to determine nitrogen (N), phosphorus (P), and potassium (K) fertilizer input rates for wheat production, and then validated the method by a field experiment at 66 different sites in northern China. The results showed that wheat grain yield varied from 1.1 to 9.2 t ha−1, averaging 4.6 t ha−1, and it had a quadratic relationship with the topsoil (0−20 cm) nitrate N and soil available P contents at harvest. However, yield was not correlated with the inputs of N, P, and K fertilizers. Based on the relationship (exponential decay model) between 95–105% of the relative yield and topsoil nitrate N, available P, and available K contents at wheat harvest from 60 field experiments, the topsoil critical nutrient values were determined as 34.6, 15.6, and 150 mg kg−1 for soil nitrate N, available P, and available K, respectively. Then, based on five groups of relative yield (>125%, 115–125%, 105–115%, 95–105%, and <95%) and the model, the five groups of topsoil critical nutrient levels and fertilization coefficients (Fc) were determined. Finally, we proposed a new method for calculating the recommended fertilizer input rate as: Fr=Gy×Nr×Fc, where Fr is the recommended fertilizer (N/P/K) input rate; Gy is the potential grain yield; Nr is the N(NrN), P(NrP), and K(NrK) nutrient requirements for wheat to produce 1,000 kg of grain; and Fc is a coefficient for N(Nc)/P(Pc)/K(Kc) fertilizer. A 2-year validated experiment confirmed that the new method reduced N fertilizer input by 17.5% (38.5 kg N ha−1) and P fertilizer input by 43.5% (57.5 kg P2O5 ha−1) in northern China and did not reduce the wheat yield. This outcome can significantly increase the farmers’ benefits (by 7.58%, or 139 US$ ha−1). Therefore, this new recommended fertilization method can be used as a tool to guide N, P, and K fertilizer application rates for dryland wheat production.
The evolution and diurnal expression patterns of photosynthetic pathway genes of the invasive alien weed, Mikania micrantha
Mikania micrantha is a fast-growing global invasive weed species that causes severe damage to natural ecosystems and very large economic losses of forest and crop production. It has advantages in photosynthesis, including a similar net photosynthetic rate as C4 plants and a higher carbon fixation capacity. We used a combination of genomics and transcriptomics approaches to study the evolutionary mechanisms and circadian expression patterns of M. micrantha. In M. micrantha, 16 positive selection genes focused on photoreaction and utilization of photoassimilates. In different tissues, 98.1% of the genes associated with photoresponse had high expression in stems, and more than half of the genes of the C4 cycle had higher expression in stems than in leaves. In stomatal opening and closing, 2 genes of carbonic anhydrase (CAs) had higher expression at 18:00 than at 8:00, and the slow anion channel 1 (SLAC1) and high-leaf-temperature 1 kinase (HT1) genes were expressed at low levels at 18:00. In addition, genes associated with photosynthesis had higher expression levels at 7:00 and 17:00. We hypothesized that M. micrantha may undergo photosynthesis in the stem and flower organs and that some stomata of the leaves were opening at night by CO2 signals. In addition, its evolution may attenuate photoinhibition at high light intensities, and enhance more efficient of photosynthesis during low light intensity. And the tissue-specific photosynthetic types and different diurnal pattern of photosynthetic-related genes may contribute to its rapid colonization of new habitats of M. micrantha.
Combined application of chemical fertilizers with organic amendments was recommended as a strategy for improving yield, soil carbon storage, and nutrient use efficiency. However, how the long-term substitution of chemical fertilizer with organic manure affects rice yield, carbon sequestration rate (CSR), and nitrogen use efficiency (NUE) while ensuring environmental safety remains unclear. This study assessed the long-term effect of substituting chemical fertilizer with organic manure on rice yield, CSR, and NUE. It also determined the optimum substitution ratio in the acidic soil of southern China. The treatments were: (i) NPK0, unfertilized control; (ii) NPK1, 100% chemical nitrogen, phosphorus, and potassium fertilizer; (iii) NPKM1, 70% chemical NPK fertilizer and 30% organic manure; (iv) NPKM2, 50% chemical NPK fertilizer and 50% organic manure; and (v) NPKM3, 30% chemical NPK fertilizer and 70% organic manure. Milk vetch and pig manure were sources of manure for early and late rice seasons, respectively. The result showed that SOC content was higher in NPKM1, NPKM2, and NPKM3 treatments than in NPK0 and NPK1 treatments. The carbon sequestration rate increased by 140, 160, and 280% under NPKM1, NPKM2, and NPKM3 treatments, respectively, compared to NPK1 treatment. Grain yield was 86.1, 93.1, 93.6, and 96.5% higher under NPK1, NPKM1, NPKM2, and NPKM3 treatments, respectively, compared to NPK0 treatment. The NUE in NPKM1, NPKM2, and NPKM3 treatments was higher as compared to NPK1 treatment for both rice seasons. Redundancy analysis revealed close positive relationships of CSR with C input, total N, soil C:N ratio, catalase, and humic acids, whereas NUE was closely related to grain yield, grain N content, and phenol oxidase. Furthermore, CSR and NUE negatively correlated with humin acid and soil C:P and N:P ratios. The technique for order of preference by similarity to ideal solution (TOPSIS) showed that NPKM3 treatment was the optimum strategy for improving CSR and NUE. Therefore, substituting 70% of chemical fertilizer with organic manure could be the best management option for increasing CSR and NUE in the paddy fields of southern China
Porcine proliferative enteropathy (PPE), an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis, is commonly associated with diarrhea and reduced weight gain in growing pigs widespread. An accurate method for detecting L. intracellularis is particularly important for preventing and controlling PPE. Heat shock protein 60 (Hsp60) is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms. Thus, the purpose of the current investigation was to produce a novel L. intracellularis Hsp60 monoclonal antibody (mAb) useful for immunodiagnostics. Three hybridomas secreted anti-Hsp60 termed 3E5, 4E2, and 9G6 were generated, and the titers of ascitic fluids of 3E5, 4E2, 9G6 were 1:1 024 000, 1:2 048 000 and 1:2 048 000, respectively. The Western blotting analysis demonstrated that recombinant Hsp60 (rHsp60) was recognized by mAbs 3E5, 4E2 and 9G6. Subsequently, analyses of specificity showed all the mAbs were highly specific to L. intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs, such as Escherichia coli, Salmonella Choleraesuis, Salmonella Typhimurium, and Brachyspira hyodysenteriae. Furthermore, the mAbs were useful for detecting L. intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs. Our research will provide a foundation for the development of immunological diagnostic tests
Anthocyanin is an important pigment that affects plant color and nutritional quality. MYBs play an important role in plant anthocyanin synthesis and accumulation. However, the regulatory function of MYB transcription factors in anthocyanin synthesis in flax flowers is still unclear. In this study, 402 MYB transcription factors were identified in the flax genome. These MYB members are unevenly distributed on 15 chromosomes. The R2R3-LuMYB members were divided into 32 phylogenetic subfamilies. qRT-PCR analysis showed that seven R2R3-LuMYB genes in the adjacent subfamily of the evolutionary tree had similar expression patterns, among which LuMYB216 was highly expressed in the petals of different colors. Moreover, gene editing of LuMYB216 in flax showed that the petal color, anther color and seed coat color of mutant plants were significantly lighter than those of wild-type plants, and the anthocyanin content of lumyb216 mutant plants was significantly reduced. Correlation analysis indicated that LuMYB216 was significantly positively correlated with the upstream regulator bHLH30. This study systematically analyzed the MYB gene family in flax, laying a foundation for studying the regulation of LuMYB216 in flax flower anthocyanin synthesis.
Maize tassel detection is essential for future agronomic management in maize planting and breeding, with application in yield estimation, growth monitoring, intelligent picking, and disease detection. However, detecting maize tassels in the field poses prominent challenges as they are often obscured by widespread occlusions and differ in size and morphological color at different growth stages. This study proposes the SEYOLOX-tiny Model that more accurately and robustly detects maize tassels in the field. Firstly, the data acquisition method ensures the balance between the image quality and image acquisition efficiency and obtains maize tassel images from different periods to enrich the dataset by unmanned aerial vehicle (UAV). Moreover, the robust detection network extends YOLOX by embedding an attention mechanism to realize the extraction of critical features and suppressing the noise caused by adverse factors (e.g., occlusions and overlaps), which could be more suitable and robust for operation in complex natural environments. Experimental results verify the research hypothesis and show a mean average precision (mAP@0.5) of 95.0%. The mAP@0.5, mAP@0.5–0.95, mAP@0.5–0.95 (area=small), and mAP@0.5–0.95 (area=medium) average values increased by 1.5, 1.8, 5.3, and 1.7%, respectively, compared to the original model. The proposed method can effectively meet the precision and robustness requirements of the vision system in maize tassel detection.
Rye (Secale cereale L., 2n=2x=14, RR) is a significant genetic resource for improving common wheat because of its resistance to multiple diseases and abiotic-stress tolerant traits. The 1RS chromosome from the German cultivated rye variety Petkus is critical in wheat breeding. However, its weakened disease resistance highlights the need to identify new resources. In the present study, a novel derived line called D27 was developed from common wheat and Mexico Rye. Cytological observations characterized the karyotype of D27 as 2n=42=21 II. Genomic in situ hybridization indicated that a pair of whole-arm translocated Mexico Rye chromosomes were inherited typically in the mitotic and meiosis stages of D27. Experiments using fluorescence in situ hybridization (FISH) and gliadin electrophoresis showed that D27 lacked wheat 1DS chromosomes. They were replaced by 1RS chromosomes of Mexico Rye, supported by wheat simple-sequence repeat markers, rye sequence characterized amplified region markers, and wheat 40K SNP array analysis. The wheat 1DS chromosomes could not be detected by molecular markers and wheat SNP array, but the presence of rye 1RS chromosomes was confirmed. Agronomic trait assessments indicated that D27 had a higher tiller number and enhanced stripe rust and powdery mildew resistance. In addition, dough properties analysis showed that replacing 1DS led to higher viscosity and lower dough elasticity in D27, which was beneficial for cake making. In conclusion, the novel cytogenetically stable common wheat–Mexico Rye T1DL·1RS translocation line D27 offers excellent potential as outstanding germplasm in wheat breeding programs focusing on disease resistance and yield improvement. Additionally, it can be valuable for researching the rye 1RS chromosome’s genetic diversity.
Investigation of Aegilops umbellulata for stripe rust resistance, heading date, and the contents of iron, zinc, and gluten protein
Aegilops umbellulata (UU) is a wheat wild relative that has potential use in the genetic improvement of wheat. In this study, 46 Ae. umbellulata accessions were investigated for stripe rust resistance, heading date (HD), and the contents of iron (Fe), zinc (Zn), and seed gluten proteins. Forty-two of the accessions were classified as resistant to stripe rust, while the other four accessions were classified as susceptible to stripe rust in four environments. The average HD of Ae. umbellulata was significantly longer than that of three common wheat cultivars (180.9 d vs. 137.0 d), with the exception of PI226500 (138.9 d). The Ae. umbellulata accessions also showed high variability in Fe (69.74–348.09 mg kg–1) and Zn (49.83–101.65 mg kg–1) contents. Three accessions (viz., PI542362, PI542363, and PI554399) showed relatively higher Fe (230.96–348.09 mg kg–1) and Zn (92.46–101.65 mg kg–1) contents than the others. The Fe content of Ae. umbellulata was similar to those of Ae. comosa and Ae. markgrafii but higher than those of Ae. tauschii and common wheat. Aegilops umbellulata showed a higher Zn content than Ae. tauschii, Ae. comosa, and common wheat, but a lower content than Ae. markgrafii. Furthermore, Ae. umbellulata had the highest proportion of γ-gliadin among all the species investigated (Ae. umbellulata vs. other species=mean 72.11% vs. 49.37%; range: 55.33–86.99% vs. 29.60–67.91%). These results demonstrated that Ae. umbellulata exhibits great diversity in the investigated traits, so it can provide a potential gene pool for the genetic improvement of these traits in wheat.
Caprine arthritis-encephalitis virus (CAEV) is an under-studied virus infecting caprines and ovines worldwide. Over the last four decades, CAEV has spread in China, obtaining genomic data on CAEV strains circulating in China is of importance for developing diagnostic methods and eradicating associated diseases. However, there is limited information on the genome, including characterizations, and the probable origin. This work aimed to characterize Chinese CAEV genomes and population structures. Five CAEV strains isolated from infected dairy goats between 1989 and 1994 in Gansu, Guizhou, Shaanxi, Shandong and Sichuan provinces were cloned and sequenced. The Chinese CAEV had a 58–93% genome similarities to strains outside of China, and they belonged to subgenotype B1. The highest similarity levels (98.3–99.3%) were with two other Chinese strains, and they shared a 91.8–92.3% similarity with the strain Clements (GenBank accession no. NC_001463.1) from outside of China. The Chinese CAEV strains isolated from different provinces over five years were still highly homologous and contained unique ancestral population components, indicating that these Chinese strains had a common origin that differed from other known strains. Our results provide genomic data on circulating Chinese CAEV strains and will be useful for future epidemiological investigations and CAEV eradication programs.
Increasing wheat yield is a long-term goal for wheat breeders around the world. Exploiting elite genetic resources and dissecting the genetic basis of important agronomic traits in wheat are the necessary methods for high-yield wheat breeding. This study evaluated nine crucial agronomic traits found in a natural population of 156 wheat varieties and 77 landraces from Sichuan, China in seven environments over two years. The results of this investigation of agronomic traits showed that the landraces had more tillers and higher kernel numbers per spike (KNS), while the breeding varieties had higher thousand-kernel weight (TKW) and kernel weight per spike (KWS). The generalized heritability (H2) values of the nine agronomic traits varied from 0.74 to 0.95. Structure analysis suggested that the natural population could be divided into three groups using 43 198 single nucleotide polymorphism (SNP) markers from the wheat 55K SNP chip. A total of 67 quantitative trait loci (QTLs) were identified by the genome-wide association study (GWAS) analysis based on the Q+K method of a mixed linear model. Three important QTLs were analyzed in this study. Four haplotypes of QFTN.sicau-7BL.1 for fertile tillers number (FTN), three haplotypes of QKNS.sicau-1AL.2 for KNS, and four haplotypes of QTKW.sicau-3BS.1 for TKW were detected. FTN-Hap2, KNS-Hap1, and TKW-Hap2 were excellent haplotypes for each QTL based on the yield performance of 42 varieties in regional trials from 2002 to 2013. The varieties with all three haplotypes showed the highest yield compared to those with either two haplotypes or one haplotype. In addition, the KASP-AX-108866053 marker of QTL QKNS.sicau-1AL.2 was successfully distinguished between three haplotypes (or alleles) in 63 varieties based on the number of kernels per spike in regional trials between 2018 and 2021. These genetic loci and reliable makers can be applied in marker-assisted selection or map-based gene cloning for the genetic improvement of wheat yield.
The red coloring of pear fruits is mainly caused by anthocyanin accumulation. Red sport, represented by the green pear cultivar ‘Bartlett’ (BL) and the red-skinned derivative ‘Max Red Bartlett’ (MRB), is an ideal material for studying the molecular mechanism of anthocyanin accumulation in pear. Genetic analysis has previously revealed a quantitative trait locus (QTL) associated with red skin color in MRB. However, the key gene in the QTL and the associated regulatory mechanism remain unknown. In the present study, transcriptomic and methylomic analyses were performed using pear skin for comparisons between BL and MRB. These analyses revealed differential PcHY5 DNA methylation levels between the two cultivars; MRB had lower PcHY5 methylation than BL during fruit development, and PcHY5 was more highly expressed in MRB than in BL. These results indicated that PcHY5 is involved in the variations in skin color between BL and MRB. We further used dual luciferase assays to verify that PcHY5 activates the promoters of the anthocyanin biosynthesis and transport genes PcUFGT, PcGST, PcMYB10 and PcMYB114, confirming that PcHY5 not only regulates anthocyanin biosynthesis but also anthocyanin transport. Furthermore, we analyzed a key differentially methylated site between MRB and BL, and found that it was located in an intronic region of PcHY5. The lower methylation levels in this PcHY5 intron in MRB were associated with red fruit color during development, whereas the higher methylation levels at the same site in BL were associated with green fruit color. Based on the differential expression and methylation patterns in PcHY5 and gene functional verification, we hypothesize that PcHY5, which is regulated by methylation levels, affects anthocyanin biosynthesis and transport to cause the variations in skin color between BL and MRB.
Psathyrostachys huashanica Keng (2n=2x=14, NsNs) is regarded as a valuable wild relative species for common wheat cultivar improvement because of its abundant beneficial agronomic traits. However, although the development of many wheat–P. huashanica-derived lines provides a germplasm base for the transfer of excellent traits, the lag in the identification of P. huashanica chromosomes in the wheat background has limited the study of these lines. In this study, three novel nondenaturing fluorescence in situ hybridization (ND-FISH)-positive oligo probes were developed. Among them, HS-TZ3 and HS-TZ4 could specifically hybridize with P. huashanica chromosomes, mainly in the telomere area, and HS-CHTZ5 could hybridize with the chromosomal centromere area. We sequentially constructed a P. huashanica FISH karyotype and idiogram that helped identify the homologous groups of introduced P. huashanica chromosomes. In detail, 1Ns and 2Ns had opposite signals on the short and long arms, 3Ns, 4Ns, and 7Ns had superposed two-color signals, 5Ns and 6Ns had fluorescent signals only on their short arms, and 7Ns had signals on the intercalary of the long arm. In addition, we evaluated different ways to identify alien introgression lines by using low-density single nucleotide polymorphism (SNP) arrays and recommended the SNP homozygosity rate in each chromosome as a statistical pattern. The 15K SNP array is widely applicable for addition, substitution, and translocation lines, and the 40K SNP array is the most accurate for recognizing transposed intervals between wheat and alien chromosomes. Our research provided convenient methods to distinguish the homologous group of P. huashanica chromosomes in a common wheat background based on ND-FISH and SNP arrays, which is of great significance for efficiently identifying wheat–P. huashanica-derived lines and the further application of Ns chromosomes
Early defoliation, which usually occurs during summer in pear trees, is gradually becoming a major problem that poses a serious threat to the pear industry in southern China. However, there is no system for evaluating the responses of different cultivars to early defoliation, and our knowledge of the potential molecular regulation of the genes underlying this phenomenon is still limited. In this study, we conducted field investigations of 155 pear accessions to assess their resistance or susceptibility to early defoliation. A total of 126 accessions were found to be susceptible to early defoliation, and only 29 accessions were resistant. Among them, 19 resistant accessions belong to the sand pear species (Pyrus pyrifolia). To identify the resistance genes related to early defoliation, the healthy and diseased samples of two sand pear accessions, namely, the resistant early defoliation accession ‘Whasan’ and the susceptible early defoliation accession ‘Cuiguan’, were used to perform RNA sequencing. Compared with ‘Cuiguan’, a total of 444 genes were uniquely differentially expressed in ‘Whasan’. Combined with GO and KEGG enrichment analyses, we found that early defoliation was closely related to the stress response. Furthermore, a weighted gene co-expression network analysis revealed a high correlation of WRKY and ethylene responsive factor (ERF) transcription factors with early defoliation resistance. This study provides useful resistant germplasm resources and new insights into potentially essential genes that respond to early defoliation in pears, which may facilitate a better understanding of the resistance mechanism and molecular breeding of resistant pear cultivars