Prescreening of large-effect markers with multiple strategies improves the accuracy of genomic prediction
Presently, integrating multi-omics information into a prediction model has become a ameliorate strategy for genomic selection to improve genomic prediction accuracy. Here, we set the genomic and transcriptomic data as the training population data, using BSLMM, TWAS, and eQTL mapping to prescreen features according to | ^βb|>0, top 1% of phenotypic variation explained (PVE), expression-associated single nucleotide polymorphisms (eSNPs), and egenes (false discovery rate (FDR)<0.01), where these loci were set as extra fixed effects (named GBLUP-Fix) and random effects (GFBLUP) to improve the prediction accuracy in the validation population, respectively. The results suggested that both GBLUP-Fix and GFBLUP models could improve the accuracy of longissimus dorsi muscle (LDM), water holding capacity (WHC), shear force (SF), and pH in Huaxi cattle on average from 2.14 to 8.69%, especially the improvement of GFBLUP-TWAS over GBLUP was 13.66% for SF. These methods also captured more genetic variance than GBLUP. Our study confirmed that multi-omics-assisted large-effects loci prescreening could improve the accuracy of genomic prediction.
Functional prediction of tomato PLATZ family members and functional verification of SlPLATZ17
PLATZ is a novel zinc finger DNA-binding protein that plays an important role in regulating plant growth and development and resisting abiotic stress. However, there has been very little research on the function of this family gene in tomatoes, which limits its application in germplasm resource improvement. Therefore, the PLATZ gene family was identified and analyzed in tomato, and its roles were predicted and verified to provide a basis for in-depth research on SlPLATZ gene function. In this study, the PLATZ family members of tomato were identified in the whole genome, and 19 SlPLATZ genes were obtained. Functional prediction was conducted based on gene and promoter structure analysis and RNA-seq-based expression pattern analysis. SlPLATZ genes that responded significantly under different abiotic stresses or were significantly differentially expressed among multiple tissues were screened as functional gene resources. SlPLATZ17 was selected for functional verification by experiment-based analysis. The results showed that the downregulation of SlPLATZ17 gene expression reduced the drought and salt tolerance of tomato plants. Tomato plants overexpressing SlPLATZ17 had larger flower sizes and long, thin petals, adjacent petals were not connected at the base, and the stamen circumference was smaller. This study contributes to understanding the functions of the SlPLATZ family in tomato and provides a reference for functional gene screening.
Phosphorus (P) is a finite natural resource and is increasingly considered to be a challenge for global sustainability. Agriculture in China plays a key role in global sustainable P management. Rhizosphere and soil-based P management are necessary for improving P-use efficiency and crop productivity in intensive agriculture in China. A previous study has shown that the future demand for phosphate fertilizer by China estimated by the LePA model (legacy phosphorus assessment model) can be greatly reduced by soil-based P management (the building-up and maintenance approach). The present study used the LePA model to predict the phosphate demand by China through combined rhizosphere and soil-based P management at county scale under four P fertilizer scenarios: (1) same P application rate as in 2012; (2) rate maintained same as 2012 in low-P counties or no P fertilizer applied in high-P counties until targeted soil Olsen-P (TPOlsen) level is reached, and then rate was the same as P-removed at harvest; (3) rate in each county decreased to 1–7 kg ha–1 yr–1 after TPOlsen is reached in low-P counties, then increased by 0.1–9 kg ha–1 yr–1 until equal to P-removal; (4) rate maintained same as 2012 in low-P counties until TPOlsen is reached and then equaled to P-removal, while the rate in high-P counties is decreased to 1–7 kg ha–1 yr–1 until TPOlsen is reached and then increased by 0.1–9 kg ha–1 yr–1 until equal to P-removal. Our predictions showed that the total demand for P fertilizer by whole China was 693 Mt P2O5 and according to scenario 4, P fertilizer could be reduced by 57.5% compared with farmer current practice, during the period 2013–2080. The model showed that rhizosphere P management led to a further 8.0% decrease in P fertilizer use compared with soil-based P management. The average soil Olsen-P level in China only needs to be maintained at 17 mg kg–1 to achieve high crop yields. Our results provide a firm basis for government to issue-relevant policies for sustainable P management in China.
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.
In situ mRNA hybridization (ISH) is a powerful tool for examining the spatiotemporal expression of genes in shoot apical meristems and flower buds of cucumber. The most common ISH protocol uses paraffin wax; however, embedding tissue in paraffin wax can take a long time and might result in RNA degradation and decreased signals. Here, we developed an optimized protocol to simplify the process and improve RNA sensitivity. We combined embedding tissue in low melting-point Steedman’s wax with processing tissue sections in solution, as in the whole-mount ISH method in the optimized protocol. Using the optimized protocol, we examined the expression patterns of the CLAVATA3 (CLV3) and WUSCHEL (WUS) genes in shoot apical meristems and floral meristems of Cucumis sativus (cucumber) and Arabidopsis thaliana (Arabidopsis). The optimized protocol saved 4–5 days of experimental period compared with the standard ISH protocol using paraffin wax. Moreover, the optimized protocol achieved high signal sensitivity. The optimized protocol was successful for both cucumber and Arabidopsis, which indicates it might have general applicability to most plants
Sex determination in plants gives rise to unisexual flowers. A better understanding of the regulatory mechanism underlying the production of unisexual flowers will help to clarify the process of sex determination in plants and allow researchers and farmers to harness heterosis. Androecious cucumber (Cucumis sativus L.) plants can be used as the male parent when planted alongside a gynoecious line to produce heterozygous seeds, thus reducing the cost of seed production. The isolation and characterization of additional androecious genotypes in varied backgrounds will increase the pool of available germplasm for breeding. Here, we discovered an androecious mutant in a previously generated ethyl methanesulfonate (EMS)-mutagenized library of the cucumber inbred line ‘406’. Genetic analysis, whole-genome resequencing, and molecular marker-assisted verification demonstrated that a nonsynonymous mutation in the ethylene biosynthetic gene 1-AMINOCYCLOPROPANE-1-CARBOXYLATE SYNTHASE 11 (ACS11) conferred androecy. The mutation caused an amino acid change from serine (Ser) to phenylalanine (Phe) at position 301 (S301F). In vitro enzyme activity assays revealed that this S301F mutation leads to a complete loss of enzymatic activity. This study provides a new germplasm for use in cucumber breeding as the androecious male parent, and it offers new insights into the catalytic mechanism of ACS enzymes.
Understanding the spatial distribution of the crop yield gap (YG) is essential for improving crop yields. Recent studies have typically focused on the site scale, which may lead to considerable uncertainties when scaled to the regional scale. To mitigate this issue, this study used a process-based and remote sensing driven crop yield model for winter wheat (PRYM-Wheat), which was derived from the boreal ecosystem productivity simulator (BEPS), to simulate the YG of winter wheat in the North China Plain from 2015 to 2019. Yield validation based on statistical yield data revealed good performance of the PRYM-Wheat Model in simulating winter wheat actual yield (Ya). The distribution of Ya across the North China Plain showed great heterogeneity, decreasing from southeast to northwest. The remote sensing-estimated results show that the average YG of the study area was 6 400.6 kg ha–1. The YG of Jiangsu Province was the largest, at 7 307.4 kg ha–1, while the YG of Anhui Province was the smallest, at 5 842.1 kg ha–1. An analysis of the responses of YG to environmental factors showed no obvious correlation between YG and precipitation, but there was a weak negative correlation between YG and accumulated temperature. In addition, the YG was positively correlated with elevation. In general, studying the specific features of the YG can provide directions for increasing crop yields in the future
Understanding the characteristics and influences of various factors on phosphorus (P) fractions is of significance for promoting the efficiency of soil P. Based on long-term experiments on black soil, fluvo-aquic soil, and loess soil, which belong to Phaeozems, Cambisols, and Anthrosols in the World Reference Base for Soil Resources (WRB), respectively, five fertilization practices were selected and divided into three groups: no P fertilizer (CK/NK), balanced fertilizer (NPK/NPKS), and manure plus mineral fertilizer (NPKM). Soil inorganic P (Pi) fractions and soil properties were analyzed to investigate the characteristics of the Pi fractions and the relationships between Pi fractions and various soil properties. The results showed that the proportion of Ca10-P in the sum of total Pi fractions was the highest in the three soils, accounting for 33.5% in black soil, 48.8% in fluvo-aquic soil, and 44.8% in loess soil. Long-term fertilization practices resulted in periodic changes in soil Pi accumulation or depletion. For black soil and fluvo-aquic soil, the Pi accumulation was higher in the late period (10–20 years) of fertilization than in the early period (0–10 years) under NPK/NPKS and NPKM, whereas the opposite result was found in loess soil. The Pi accumulation occurred in all Pi fractions in black soil; mainly in Ca8-P, Fe-P, and Ca10-P in fluvo-aquic soil; and in Ca2-P, Ca8-P, and O-P in loess soil. Under CK/NK, the soil Pi was depleted mainly in the early period in each of the three soils. In addition to the labile Pi (Ca2-P) and moderately labile Pi (Ca8-P, Fe-P, Al-P), the Ca10-P in black soil and fluvo-aquic soil and O-P in loess soil could also be used by crops. Redundancy analysis showed that soil properties explained more than 90% of the variation in the Pi fractions in each soil, and the explanatory percentages of soil organic matter (SOM) were 43.6% in black soil, 74.6% in fluvo-aquic, and 38.2% in loess soil. Consequently, decisions regarding the application of P fertilizer should consider the accumulation rate and the variations in Pi fractions driven by soil properties in non-acidic soils.
Trichomes are specialized structures developed from epidermal cells and can protect plants against biotic and abiotic stresses. Trichomes cover carrots during the generative phase. However, the morphology of the carrot trichomes and candidate genes controlling the formation of trichomes are still unclear. This study found that carrot trichomes were non-glandular and unbranched hairs distributed on the stem, leaf, petiole, pedicel, and seed of carrot. Resequencing analysis of a trichome mutant with sparse and short trichomes (sst) and a wild type (wt) with long and dense trichomes on carrot stems was conducted. A total of 15 396 genes containing nonsynonymous mutations in sst were obtained, including 42 trichome-related genes. We also analyzed the transcriptome of the trichomes on secondary branches when these secondary branches were 10 cm long between wt and sst and obtained 6 576 differentially expressed genes (DEGs), including 24 trichome-related genes. qRT-PCR validation exhibited three significantly up-regulated DEGs, 20 significantly down-regulated, and one with no difference. We considered both the resequencing and transcriptome sequencing analyses and found that 12 trichome-related genes that were grouped into five transcription factor families containing nonsynonymous mutations and significantly down-regulated in sst. Therefore, these genes are potentially promising candidate genes whose nonsynonymous mutations and down-regulation may result in scarce and short trichomes mutation on carrot stems in sst.
Peel color is an important appearance quality of melons that greatly affects consumer preferences. In this study, a near-isogenic line NIL-G (dark green peel) was generated from B8 (grey-green peel) and B15 (white peel). The F2 population constructed by crossing NIL-G and B15 was used to study the inheritance pattern of peel color, and bulked-segregant analysis sequencing (BSA-seq) was employed to identify the interval in which the target gene was located. Genetic analysis results showed that the dark green peel trait at maturity is controlled by a dominant gene. BSA-seq and molecular markers were used to localize the candidate gene in a 263.7 kb interval of chromosome 4, which contained the CmAPRR2 gene with known functions. Moreover, allelic sequence analysis revealed four SNP variations of the CmAPRR2 gene in B15, of which SNP.G614331A was located at the junction of the 6th exon and 6th intron. The G-to-A mutation caused alternative splicing of the transcript of CmAPRR2 in B15, generating two transcripts (CmAPRR2-A and CmAPRR2-B) with premature termination codons. Furthermore, the Kompetitive Allele Specific PCR (KASP) marker, APRR2-G/A, was developed based on this SNP and shown to co-segregate with the peel color phenotype in the F2 population. Compared to white-peel B15, the expression level of CmAPRR2 in dark green peel NIL-G was higher at each growth stage. Therefore, CmAPRR2 may be the key gene controlling the fruit color of melons. Overall, this study identified a novel allelic variant of CmAPRR2 that leads to white peel formation in mature melons. The study also provides a theoretical basis for further research on the gene regulatory mechanism of melon peel colors and may promote the future use of molecular marker-assisted selection to modify melon peel colors.