【Objective】 The heading date plays a crucial role in influencing the regional adaptation and yield of rice (Oryza sativa L.). The identification of early heading genes contributes significantly to enhancing and fine-tuning the regulatory network that controls rice heading, which provides valuable genetic resources for molecular breeding with the goal of achieving early maturity and high yield in rice. 【Method】 CL33, a chromosome segment substitution line with early heading, and 93-11, its recipient parent with late heading, were used as research materials to investigate and analyze their major agronomic traits. Two DNA pools were constructed, comprising plants exhibiting extremely early and late heading. Whole-genome resequencing and BSA-Seq analyses were then conducted to locate the genomic region associated with the heading date. In the subsequent steps, InDel markers within this identified region were developed for fine mapping. The gene LOC-Os08g07740 emerged as the primary candidate gene within localization intervals, determined through gene prediction, candidate gene analyses, as well as references to relevant literatures. This candidate gene was subsequently cloned and analyzed for allelic variations. Moreover, we explored the genetic and phylogenetic relationships of the LOC_Os08g07740 gene within the three rice subgroups, Indica, Japonica and O. rufipogon. This analysis involved studying genomic data within approximately 40 kb upstream and downstream of the gene utilizing bioinformatics software.【Result】 Under both natural long-day and short-day conditions in Nanning, Guangxi, CL33 exhibited a 20-25 days shorter than its recipient parent 93-11. Moreover, under natural long-day conditions, the agronomic traits of CL33 were largely similar to those of 93-11, with the exception of a shortened spike length and a reduced number of grains per spike. Genetic analysis revealed that the early heading trait in CL33 was controlled by a recessive gene. This gene was finely localized within a 100 kb region between the markers PSM8-6 and PSM8-8 on the short arm of rice chromosome 8, encompassing 15 predicted candidate genes. Significantly, the candidate gene ORF13 (LOC_Os08g07740), which shared alleles with known heading date genes like DTH8/Ghd8 emerged as a key candidate. Sequencing and sequence alignment of ORF13 demonstrated an 888 bp coding sequence without introns, encoding a protein of 295 amino acids. Compared to the recipient parent 93-11, LOC_Os08g07740 in CL33 featured a 6 bp insertion and a 9 bp deletion between the 535-536th and 820-821st base pairs, respectively, resulting in consequential amino acid sequence alterations. Hence, it was tentatively named OsEHD8 as the target candidate gene. Genetic evolutionary analyses indicated a significant decrease in genetic diversity within the LOC_Os08g07740 gene in Indica and Japonica compared to O. rufipogon, with a 62.53% decrease in Indica and a 53.76% decrease in Japonica. Nevertheless, the differences in genetic diversity between Indica and Japonica were not statistically significant. The LOC_Os08g07740 gene featured a total of 13 haplotypes, with the Hap_2 possibly representing the common ancestor of the three subgroups. Geographical isolation or environmental differences might have led to the fixation of different haplotypes in the Indica and Japonica subgroups. These findings suggested that the LOC_Os08g07740 gene underwent directional selection in the three subgroups.【Conclusion】 OsEHD8, identified as a functional allele of DTH8/Ghd8, played a key role in promoting early heading in rice under both natural long-day and short-day conditions. Moreover, the chromosomal segment substitution line CL33, which carried the OsEHD8 allele, exhibited no significant differences in other agronomic traits compared to the recipient parent 93-11 under natural long-day conditions, except for a shorter spike length and a reduction in grains per spike.

【Objective】 Enolase is a key rate-limiting enzyme in the process of glycolysis, and plays a crucial role in plant growth and development, and response to abiotic stress. The function of common wheat enolase gene TaENO1-5B was revealed and molecular markers were developed to provide genetic resources for improving wheat through molecular breeding. 【Method】 TaENO1-5B was cloned from wheat variety Hanxuan 10. The domains of its encoded protein were analyzed on the SMART website. The secondary and tertiary structures of the protein were predicted by Phyre2 software. Real-time quantitative fluorescent PCR was used to analyze the expression levels of the target gene in wheat tissues at different developmental stages and its expression patterns under phytohormone treatment and abiotic stress. Thirty wheat germplasm with rich genetic diversity were used as plant materials to analyze the gene sequence polymorphisms, and develop molecular markers. The association analysis between TaENO1-5B haplotypes and phenotypic traits was carried out in a natural population consisting of 323 wheat accessions. The trend of breeding selection of superior haplotype in different wheat production zones in China was analyzed by using a landrace population and a modern variety population. 【Result】 TaENO1-5B gene consists of 17 exons and 16 introns encoding 446 amino acids and contains a conserved N-terminal domain and a C-terminal TIM (triose-phosphate isomerase) barrel domain. TaENO1-5B was expressed in all tissues of wheat at seedling, jointing, heading and flowering stages, and the expression level was higher in roots, root bases and spikes. The TaENO1-5B promoter region contains a variety of cis-acting elements, including elements responding to plant hormones, such as abscisic acid (ABA), auxin (IAA), and methyl jasmonate (MeJA), as well as elements responding to drought and low temperature. The expression of TaENO1-5B was significantly induced by phytohormones and abiotic stress in wheat. Four SNPs were detected in the promoter region and three SNPs in the gene region of the TaENO1-5B gene, which constituted three haplotypes, i.e., Hap-5B-1, Hap-5B-2, and Hap-5B-3. Among them, Hap-5B-2 was a favorable haplotype highly associated with shorter plant height, more spikelets per spike, and more grains per spike under various environments such as drought and high temperature, and had been positively selected in the breeding history of major wheat production zones in China. The KASP (kompetitive allele-specific PCR) marker developed based on the SNP (2 399 bp, G/A) of the TaENO1-5B promoter region was significantly correlated with the spikelet number per spike in multiple environments. 【Conclusion】 TaENO1-5B gene responds to phytohormones and abiotic stress, and is significantly correlated with plant height, spikelet number per spike and grain number per spike under various environments such as drought and high temperature. Hap-5B-2 is a favorable haplotype with shorter plant height and more number of spikelets and grains per spike. Molecular markers developed based on the variation sites of TaENO1-5B gene sequence can be used for genetic improvement of plant height and related yield traits in wheat.

Maize is the most widely cultivated, used and highest yield crop in the world and China. Southern corn rust (SCR) is an air borne disease caused by Puccinia polysora Underw., which mainly occurs in tropical and subtropical maize growing areas. In recent years, SCR has become one of the major diseases in the Huang-Huai-hai maize production region due to the climate change, which directly leads to compromised grain quality and poor yields in maize and significantly jeopardizes maize production in China. At present, SCR usually spreads in a large area within a short period of time once occurred because most maize varieties promoted in China are susceptible, and conventional chemical measures is usually in vain. Therefore, cultivating resistant cultivars by exploiting resistance genes in maize germplasm resources is the most effective and economical strategy for controlling SCR. The highly resistant germplasm is scarce in maize resources, mainly from tropical and subtropical regions, and barely no temperate germplasm can be directly used in breeding practice. Compared with foreign maize germplasm, the highly resistant maize germplasms of China were much less, mainly from local landraces or P group materials containing tropical origins with relatively limited genetic variation. The identification and cloning of SCR resistance genes in maize is essential for promoting molecular marker-assisted breeding, as well as accelerating the breeding process of new varieties with desired resistance. At present, several SCR resistance genes have been identified and cloned, laying a foundation for molecular marker-assisted selection. Over the years, Chinese breeders have developed a number of elite maize inbred lines resistant to SCR with limited resistance germplasm resources, and successfully created disease-resistant hybrids. Recent studies on the genome of SCR pathogens revealed that pathogens have differentiated into highly toxic lineages in China, thus escaping the recognition of resistance genes. Therefore, the exploration and utilization of extensive genetic resources in resistant germplasm still need to be further strengthened. In this paper, we outlined the biological characteristics and hazards of SCR, systematically summarized the research progresses in the identification and utilization of maize germplasm resources resistant to SCR, the mapping and cloning of SCR resistant genes and the breeding of resistant varieties, and prospect the future research direction of SCR. This review will provide references for the prevention and control of SCR, as well as the breeding of resistant maize varieties.

【Objective】 This study aimed to investigate the effects of row spacing and seeding rate on forage yield of Elymus sibiricus (E. sibiricus) in alpine region, further to screen out the optimum planting row spacing and seeding rate for forage production of E. sibiricus in the Qinghai-Tibet Plateau, and to reveal the influence process and path coefficient of planting row spacing and seeding rate on forage yield of E. sibiricus, and hence, so as to provide the data support for efficient forage production in the Qinghai-Tibet Plateau. 【Method】 In this study, E. sibiricus cv. Qingmu No. 1 and E. sibiricus cv. Qingmu No. 2 were selected as experimental material, and the two-factor split zone experimental design was adopted. Three row spacings were set as the main factors, including R₁: 15 cm, R₂: 30 cm, and R₃: 45 cm, respectively, and three seeding rates (S₁: 15.0 kg·hm^-2, S₂: 22.5 kg·hm^-2, and S₃: 30.0 kg·hm^-2) were used as secondary factors. And thus, there were 9 treatments, 3 repetitions, 27 plots of each variety, and a total of 54 plots with each area 15 m²(3 m ×5 m), which was carried out in Gonghe County, Qinghai Province in 2022-2023. Random block arrangement was used to analyze the effects of different row spacings and seeding rates on growth character, forage yield and economic benefit of E. sibiricus, and the structural equation model was performed to study the influence process of row spacing and seeding rate on forage yield of E. sibiricus and its path coefficient. 【Result】 Both row spacing and seeding rate had significant effects on stem diameter, tillers, fertile tillers per, stem-leaf ratio and forage yield of Qingmu No. 1 and Qingmu No. 2. Under the same row spacing, the plant height and stem diameter of Qingmu No. 1 and Qingmu No. 2 decreased with the increasing of seeding rates, tillers and fertile tillers increased first and then decreased with the increasing of seeding rates. Under the same seeding rates, the plant height, stem diameter and dry- fresh ratio of Qingmu No. 1 and Qingmu No. 2 increased with the increasing of row spacing, and the tillers and fertile tillers decreased with the increasing of row spacing. Hence, the row spacing was 15 cm and the seeding rate was 22.5 kg·hm^-2, the forage yield and economic benefit of Qingmu No. 1 and Qingmu No. 2 were the highest, the forage yield of Qingmu No.1 was 12 668.16 kg·hm^-2, and the economic benefit was 13 731.97 yuan·hm^-2. The forage yield of Qingmu No. 2 was 12 180.94 kg·hm^-2, and the economic benefit was 13 064.03 yuan·hm^-2. Pearson correlation analysis showed that forage yield was positively correlated with tillers and fertile tillers. The structural equation model showed that tillering was the primary driving factor for improving the forage yield of E. sibiricus under row spacing and seeding rate. 【Conclusion】 The comprehensive evaluation of TOPSIS model showed that the row spacing was 15 cm and the seeding rate was 22.5 kg·hm^-2, which could not only maintain higher forage yield, but also significantly improve economic benefits, so it was the best row spacing and seeding rate amount, which was suitable for the forage production of E. sibiricus in chernozem of alpine region.

【Objective】 Exploring the impact of intercropping on the quality of Mesona chinensis and its mechanism of action, providing a theoretical basis for the development of high-quality cultivation techniques for Mesona chinensis. 【Method】 A field randomized block experiment was conducted, including three cropping systems, such as soybean/Mesona chinesi/corn intercropping (S/M/C), soybean/Mesona chinesi intercropping (S/M), and Mesona chinesi monoculture (M). The effects of different intercropping systems on the quality of Mesona chinensis and its rhizosphere soil characteristics were analyzed. 【Result】 The S/M/C and S/M intercropping were beneficial for promoting the accumulation of trace elements in Mesona chinensis leaves and stems, such as Ca (11.36%-24.20% in leaves and 33.44%-38.16% in stems), Mg (34.41%-52.00% in leaves and 15.20%-91.99% in stems), Fe (15.21%-15.46% in leaves), and Cu (17.19%-30.73% in stems). The S/M/C intercropping significantly increased the flavonoid content in the stems of Mesona chinensis by 44.42%. The two intercropping systems significantly reduced the nutrient contents of total nitrogen (TN), alkaline nitrogen (AHN), and available potassium (AK) in the rhizosphere soil of Mesona chinensis, which significantly improved the soil pH (4.82 in M, 5.22 in S/M, and 5.51 in S/M/C). However, pH was the most important soil factor driving changes in bacterial community structure in this study. The S/M/C intercropping significantly improved the bacterial diversity in the rhizosphere soil of Mesona chinensis. The two intercropping systems significantly increased the relative abundance of the dominant bacterial genus Bacillus, from 3.24% (M) to 5.28% (S/M) and 8.09% (S/M/C), respectively. In addition, the S/M/C intercropping promoted the recruitment of more bacterial phyla, such as Bdellovibrionota, Dependentiae, WS2, as well as bacterial genera such as Metrocystis in the rhizosphere soil of Mesona chinensis.【Conclusion】 The S/M/C intercropping could promote the accumulation of flavonoids in Mesona chinensis, which was beneficial for improving its quality. The improvement of soil pH might be the main factor driving the change of soil bacterial diversity and community structure in the S/M/C intercropping. The enrichment of specific bacterial phyla and genera in the rhizosphere soil of Mesona chinensis in S/M/C intercropping were beneficial for improving the biological characteristics of soil. Therefore, the reasonable intercropping such as S/M/C was an effective measure to achieve high-quality cultivation of Mesona chinensis.

【Background】 Grapevine berry inner necrosis virus (GINV) is a positive single-stranded RNA virus reported in China in recent years, which is widespread and harmful. Highly sensitive detection technology is the key for field monitoring of the virus and the cultivation of virus-free seedlings.【Objective】 The objective of this study is to establish a high-sensitivity reverse transcription real-time quantitative PCR (RT-qPCR) detection system for GINV, clarify the sensitivity of different grape rootstocks to GINV, and to clarify the spatial and temporal distribution of GINV in host plants, so as to provide technical support for the monitoring and early warning of the virus.【Method】 Six sets of primers were designed according to the conserved sequences of replicase (RP), movement protein (MP) and coat protein (CP) genes registered in GenBank. The primers with strong specificity and good amplification effect were screened by conventional RT-PCR and RT-qPCR. Then, the annealing temperature and concentration of the primers were optimized to establish the SYBR Green I dye RT-qPCR detection system for GINV, and the sensitivity, specificity and field applicability of this system were further evaluated. GINV was inoculated into five grape rootstocks, including Beta, SO4, 101-14, 140R and 1103P, for symptom observation and virus detection to screen indicator plants with high GINV sensitivity. Based on the established RT-qPCR technology, samples of different grape rootstocks inoculated with GINV were detected at different stages and different parts, so as to clarify the spatial and temporal distribution of GINV in different grape rootstocks.【Result】 The SYBR Green I dye RT-qPCR detection technology system for GINV was established, and the optimal primer was GINVRPYGF2/R2 and the optimal primer concentration was 300 nmol·L^-1, the optimal annealing temperature was 58.4 ℃. This technique had strong specificity and high sensitivity for GINV detection, and its detection sensitivity was 1 000 times that of conventional RT-PCR. The observation results of GINV inoculation into different grape rootstocks showed that Beta was the most severe in GINV infection, which was manifested as systemic necrosis of leaves, while the leaves of other rootstocks only showed the symptoms of green mottling and ring spots. The results of RT-qPCR showed that the relative content of GINV was the highest in EL27 (setting stage), and there was no significant difference in the relative content of GINV among the five varieties during EL12 (inflorescence clear stage) and EL27, and there were significant differences in the relative content of GINV between Beta and SO4, 101-14, 140R and 1103P in EL31 (berries pea size stage), and the relative content of GINV varied greatly in different tissues, and the order from high to low was lower leaves, upper leaves, upper stems, lower stems and roots.【Conclusion】 A high-sensitivity and strong-specificity RT-qPCR method for the detection of GINV was established, and the spatial and temporal distribution of GINV in different grape rootstocks was clarified by this method.

【Objective】 This study aims to establish a novel technique for detecting the west African strain of sweet potato chlorotic stunt virus (SPCSV-WA) by combining reverse transcriptase recombinase polymerase amplification and lateral flow dipstick (RT-RPA-LFD).【Method】 Primers and probes with good amplification results and strong specificity were designed and screened on the basis of the conserved sequences of the SPCSV-WA coat protein gene and heat shock protein gene. Then, the conditions such as primer and probe concentrations, amplification system, temperature, and reaction time were optimized to detect SPCSV-WA using RT-RPA-LFD. This method was used to detect common viruses on sweet potato such as the east African strain of sweet potato chlorotic stunt virus (SPCSV-EA), sweet potato feathery mottle virus (SPFMV), sweet potato latent virus (SPLV) and sweet potato virus G (SPVG) to verify the specificity. Total RNA from SPCSV-infected sweet potato leaves was diluted in a 10-fold gradient, and RT-PCR and RT-RPA-LFD were performed to compare the sensitivity of the two methods. Field sweet potato samples and test tube seedling samples were subjected to RT-RPA, RT-RPA-LFD, and RT-PCR detection to validate the practicality of the method.【Result】 The RT-RPA-LFD detection method for SPCSV-WA was established using the optimal primer CSV357F/R and the CSV-CP-Probe (47 bp). The working concentrations of the primer and probe were 0.2 and 0.06 μmol·L^-1, respectively. The reaction conditions were set to 42 ℃ for 5 min. This method could specifically detect SPCSV-WA and had no cross-reaction with other common viruses on sweet potato. The RT-RPA-LFD could detect viruses up to 10^-4 dilution, whereas RT-PCR could only detect up to 10^-3 dilution, making the sensitivity of RT-RPA-LFD 10 times higher than that of RT-PCR. Of the 22 field-gathered sweet potato samples tested by RT-PCR, RT-RPA, and RT-RPA-LFD, 11 positive samples were consistently found across the three methods. The testing results of 28 test tube seedling samples showed that RT-PCR and RT-RPA-LFD consistently detected five positive samples.【Conclusion】 The RT-RPA-LFD detection method for SPCSV-WA has been established and it is characterized by its speed, simplicity, specificity, sensitivity, and visibility. This method can be used for testing virus-free test tube seedling samples of sweet potato as well as for on-site rapid detection of field sweet potato samples in basic level units.

【Objective】 The aim of this study was to explore the effects of green manure return to the field on the content of soil organic carbon components and the activities of carbon invertase enzymes in dry-crop wheat fields, so as to provide data support for the improvement of soil quality and the achievement of the goal of “carbon neutrality”. 【Method】 The green manure return field experiment in a 5-year rotation system of hairy vetch (Vicia villosa Roth)-winter wheat (Triticum aestivum L.) and forage rape (Brassica napus L.)-winter wheat was conducted on a typical black clay soil in the Longdong dry loess area of Gansu Province. Soil organic carbon (SOC), easily oxidizable organic carbon (EOC), microbial biomass carbon (MBC) content and β-1,4-glucosidase (βG), cellobiose hydrolase (CBH), β-xylosidase (βX) geometric mean enzyme activity (GMEA) activities were analyzed in four soil layers of different stages of winter wheat after the mulching and overturning of hairy vetch, forage rape. 【Result】 The method of green manure returning to the field had a significant effect on the content of soil organic carbon components and the activities of CBH and βX. Compared with mulching, hairy vetch and forage rape overturning were able to increase the content of soil SOC, EOC and MBC by 12.9%, 12.1% and 53.8%, while the activity of βG, CBH increased by 3.2% and 10.2%, respectively, and the most significant effect was found in the 20-25 cm soil layer. There were significant differences in soil labile organic carbon content and soil enzyme activities at different winter wheat growth period, in which soil EOC and MBC content reached the highest at the maturity and greening stage of winter wheat, respectively, and the activities of βG, CBH, βX and GMEA reached the highest at the booting stage of winter wheat. The soil βG activity had the most significant changes and highest in different return methods and it was the main enzyme participating in the soil carbon transformation process after green manure returned to the field. Soil carbon component content and carbon invertase enzyme activity differed significantly in different soil layers, and both decreased with the increase of soil depth. The type of green manure also significantly affected soil carbon components and enzyme activities, in which the SOC and MBC content and the activities of soil βG, CBH, βX and GMEA returned to the field by forage rape were 1.08, 1.21, 1.15, 1.23, 1.19, and 1.19 times higher than common vetch. Structural equation modeling indicated that the green manure return method affected SOC accumulation by regulating the cumulative decomposition rate, and could regulate soil pH, SOC and the accumulated decomposition rate of green manure affecting GMEA activity. SOC accumulation was more affected by the green manure return biomass more than return method, while the opposite was true for carbon invertase enzyme activity.【Conclusion】 Cultivating and overturning forage rape during summer fallow period significantly increased soil organic carbon components and carbon invertase enzyme activities in 0-25 cm soil layer, which was an effective measure for efficient resource utilization during summer fallow period in Loess Plateau.

【Objective】 This work aimed to investigate the impact of long-term applying organic and inorganic fertilizers on the abundance and activities of complete ammonia oxidizers (Comammox), ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as the abundances of Nitrospira and Nitrobacter in paddy soil, providing a scientific basis for mitigating greenhouse gas emissions and promoting sustainable agricultural production.【Method】 By utilizing multiple specific inhibitors (i.e., acetylene, 1-octyne, and DMPP) in conjunction with real-time quantitative PCR (qPCR), this work examined the differences in activities and abundances of Comammox, AOA, and AOB as well as the abundances of Nitrobacter and Nitrospira in a paddy soil under four fertilization regimes: plots without fertilizer (CK), with manure (M), with chemical fertilizer (NPK), and with combination of manure and chemical fertilizer (MNPK).【Result】 The long-term application of organic fertilizer significantly stimulated the activities of Comammox and AOA (Two-way ANOVA, P<0.001). In those plots solely receiving organic manure, Comammox accounted for as high as 64.2% of total ammonia oxidizing activity, while the inorganic fertilizer application only enhanced the activity of AOB (Two-way ANOVA, P<0.001). qPCR demonstrated that chronic organic amendment significantly increased the abundances of AOA, Nitrospira, Comammox clade A and Clade B (Two-way ANOVA, P<0.001); whereas inorganic amendment increased the abundances of AOB and Nitrobacter (Two-way ANOVA, P<0.001). The correlation analysis revealed there were positive correlations between activities of AOA and Comammox with moisture content, organic matter (OM), total nitrogen (TN), total phosphorus (TP), available phosphorus (AP), alkaline hydrolysable nitrogen (AN), as well as AOA and Nitrospira abundances, while the activity of Comammox was positively correlated with the abundance of Comammox clade A as well. Additionally, the activity of AOB showed positive correlations with AOB and Nitrobacter abundances, nitrate content, and available potassium (AK).【Conclusion】 Comammox played an important role in nitrification of the tested paddy soil, with its abundance and activity primarily influenced by the changes in moisture content, OM, TN, TP, AP, and AN etc..

【Objective】 The objective of this study was to clarify the response of soil available zinc (Zn) and wheat grain Zn concentration to soil Zn fertilization under different Zn supply field conditions, and to explore the Zn fertilizer regulation measures for grain Zn fortification based on soil available Zn, so as to provide a scientific basis for optimizing Zn fertilizer application and achieving wheat grain with high-yield and high-quality. 【Method】 The two-year location-fixed field experiments with five Zn fertilizer application rates of 0, 6, 12, 18, and 24 kgZn·hm^-2 were carried out at Taigu (high-Zn field) and Wanrong (low-Zn field) of Shanxi Province in the eastern Loess Plateau, respectively. The wheat grain yield and Zn concentration, Zn uptake and its translocation and distribution in the aerial part, as well as soil available Zn were investigated in the high- and low-Zn fields. 【Result】 Grain yield was not affected by Zn fertilizer rates in both high- and low-Zn fields. In high-Zn field, a slight increase in grain Zn concentration was observed with the increase of Zn fertilizer rates. For grain Zn concentration, no significant difference existed among all treatments in the first year, while it was increased by 2.4%-11.0% for Zn fertilization treatments as compared with that of no Zn fertilization in the second year. The grain Zn concentration was higher than 40 mg·kg^-1 for all treatments. Compared with Zn application, the Zn transfer factor from straw to grain and grain Zn portioning index were decreased by 23.9%-37.9% and 4.3%-13.1%, respectively, and more than 20% of shoot Zn still remained in the stems and leaves at wheat harvest. In low-Zn field, the grain Zn concentration and Zn uptake in each organ increased with increasing Zn rates, whereas the opposite trend was observed for Zn transfer factor from straw to grain. Compared with no Zn application, the grain Zn concentration averaged two years increased by 9.4%-23.1%, while the Zn transfer factor from straw to grain decreased by 13.5%-24.5%, but no obvious difference was found for Zn portioning index among five Zn rates. In both high- and low-Zn fields, the soil available Zn increased significantly with the added Zn fertilizer. The regression analysis showed that soil available Zn slightly increased grain Zn concentration, and the increase with available Zn could be described by a quadratic function in high-Zn field, and the linear-with-plateau model showed that the grain Zn plateau of 34.76 mg·kg^-1 was reached at the soil available Zn of 4.12 mg·kg^-1. 【Conclusion】 Therefore, for the purpose of achieving desirable grain Zn concentration of 40 mg·kg^-1 in the wheat monoculture aera of eastern Loess Plateau, it could be considered that higher soil available Zn played a critical role in the high-Zn field, and soil Zn fertilization could be considered to increase soil available Zn up to 4 mg·kg^-1 first, and then other agronomic measures such as foliar Zn application should not be ignored to address the gap between the current grain Zn concentration and the recommended value in the low-Zn field.

【Objective】 This study aimed to explore the changes in the transcriptional level and physiological responses of pepper (Capsicum annuum L.) seedlings under different phosphorus stress gradients, to analyze the key pathways, and to integrate related experiments to elucidate the physiological mechanisms of the pepper response to nutritional stress of phosphorus. This study also identified the transcription factors and core genes that regulate the nutritional stress of phosphorus, so as to provide a theoretical basis and genetic resources for the breeding of peppers. 【Method】 This study used the roots of the pure CA#8 seedlings of pepper as experimental material, grown by Hoagland hydroponics in a single four-leaf stage and subjected to four different gradient phosphorus stress treatments, namely control group (CK, 200 μmol·L^-1 NH₄H₂PO₄) and phosphorus deficiency group (DP, 0 μmol·L^-1 NH₄H₂PO₄), low phosphorus stress group (LP, 20 μmol·L^-1 NH₄H₂PO₄), and high phosphorus stress group (HP, 1 000 μmol·L^-1 NH₄H₂PO₄). Transcriptome sequencing (RNA-Seq) was performed on pepper roots systems 2 days after treatment, and the endogenous hormone levels and antioxidant enzyme activities were measured after 0, 2, 4, and 6 days of treatment, respectively. 【Result】 Compared with the CK group, the DP, LP, and HP groups exhibited 626, 107, and 171 differentially expressed genes (DEG), respectively. 10 DEGs related to the phosphorus signaling pathway, 4 DEGs related to the plant hormone signal transduction pathway, and 7 DEGs related to antioxidant enzyme activities were identified by GO, KEGG enrichment analysis, and WGCNA analysis. Real-time fluorescence quantitative (qRT-PCR) was used to validate the accuracy of transcriptome data. Quantitative measurements of endogenous hormones in the roots of pepper seedlings showed that with increasing duration phosphorus stress, the levels of various growth-promoting endogenous hormones such as gibberellin (GA), brassinolide (BR), cytokinin (CTK), indole acetic acid (IAA), strigolactone (SL), and jasmonic acid (JA) decreased in the DP, LP and HP groups, while the levels of growth-inhibiting hormones such as ethylene (ETH) and abscisic acid (ABA) increased, especially under phosphorus deficiency and low phosphorus stress, where growth inhibition of the seedling roots was the most significant. Various phosphorus stress-induced increased activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in pepper root tissue, while the activities of SOD decreased at the later stage of stress, and POD and CAT tended to be stable too. 【Conclusion】 During various phosphorus nutritional stress, the pepper seedlings mitigated the effects of phosphorus stress on growth by responding to phosphorus signaling pathways, plant hormone signal transduction pathways, and differentially expressed genes related to antioxidant enzyme activity, thus improving the tolerance to phosphorus stress of pepper seedlings.

【Objective】 This study aimed to explore the effects of different rootstocks on the growth and fruit quality of Ruixianghong apple tree with multi-stem shape, which could provide a theoretical basis for the production of Ruixianghong apple with suitable rootstock and multi-stem shape. 【Method】 The three years old of Ruixianghong/Qingzhen 1, Ruixianghong/Malus prunifolia var. ringo, and Ruixianghong/M9T337 with multi-stem shape were used as test materials. The tree height, branch composition, annual branches growth, photosynthetic characteristics, mineral element content and fruit quality of different rootstock-scion combinations were detected and analyzed from April 2023 to March 2024. 【Result】 The results demonstrated that the tree height, graft compatibility, branch numbers, annual branch growth and photosynthetic characteristics of Ruixianghong/Malus prunifolia var. ringo were significantly higher than that of Ruixianghong/Qingzhen 1 and Ruixianghong/M9T337, with a relatively vigorous growth of vegetative organs. The growth potential of Ruixianghong/Qingzhen 1 was slightly lower than that of Ruixianghong/Malus prunifolia var. ringo, while higher than Ruixianghong/M9T337, which could better maintain the fruit shape, with the higher fruit hardness and storage quality, but the fruit yield declined, and the organic acid content increased. Ruixianghong/M9T337 had higher proportion of short branches, which balanced the vegetative growth of each part of the canopy, leading to higher yield and better fruit quality. There were apparent differences of the mineral element content in the new shoot leaves of three kinds of rootstock-scion combinations during spring and autumn shoot growing periods. The content of N, Mg, Fe, B in leaves of Ruixianghong/Malus prunifolia var. ringo and Ruixianghong/Qingzhen 1 were significantly higher than that of Ruixianghong/M9T337, and the Mn and Zn content in leaves of Ruixianghong/Malus prunifolia var. ringo and Ruixianghong/ Qingzhen 1 leaves were significantly higher than that of Ruixianghong/M9T337 during spring shoot growing period, while the content of P, K, Mo, and Cu in Ruixianghong/M9T337 leaves were higher than the other two rootstock-scion combinations, while Mn and Zn contens in leaves of Ruixianghong/M9T337 were also higher than the other two rootstock-scion combinations. 【Conclusion】 In summary, Ruixianghong/M9T337 combination with multi-stem shape had the best dwarfing effect, with higher yield and better fruit quality, but the tree potential was lower, with a reduced stress tolerance. The Ruixianghong/Malus prunifolia var. ringo combination had higher graft compatibility, growing potential, with a better tree density, stress tolerance and fruit quality in combined with the multi-stem shape.

【Background】 Accurate detection of sodium (Na), potassium (K) and magnesium (Mg) content in milk contributes to healthy dairy farming and is a prerequisite for stabilizing the quality of dairy products. However, the current conventional methods for detecting mineral content in milk are expensive and time-consuming, so there is a need for a low-cost and rapid method to measure the Na, K and Mg content in milk. 【Objective】 The purpose of this study was to investigate the potential of using milk-infrared spectroscopy (MIRS) to predict the Na, K and Mg content in milk from Chinese Holstein cows, to provide a rapid detection technique for the determination of Na, K and Mg content in milk, and to provide a large amount of phenotypic data for the herd management and genetic breeding of dairy cows. In addition, the ability of different feature selection algorithms to improve the MIRS quantitative prediction model for predicting Na, K and Mg content in milk were compared. 【Method】 A total of 255 milk samples from healthy Holstein cows from North China were used for this study. Firstly, MIRS data of milk samples were collected using MilkoScanTMFT+, and the true values of Na, K and Mg content in milk samples were determined using inductively coupled plasma atomic emission spectrometry. Subsequently, using the MIRS data as the predictor variables and the true values of Na, K and Mg content as the dependent variables, four spectral preprocessing algorithms (first-order derivative, second-order derivative, SG smoothing, and standard normal transform), four feature selection algorithms [uninformative variable elimination (UVE), competitive adaptive reweighted sampling (CARS), genetic algorithm, and least angle regression (LAR)] and nine modelling algorithms (partial least squares regression, support vector machines, Random Forest and Elasticity Network, etc.) were used to establish MIRS quantitative prediction models for predicting Na, K and Mg content in milk, respectively, and the optimal model combination (Feature Selection Algorithm + Spectral Preprocessing Algorithm + Modelling Algorithm) was selected. 【Result】 Overall, the CARS algorithm improved the Na, K and Mg content prediction models better than the UVE, GA and LAR algorithm. The Na content prediction model developed based on CARS feature selection algorithm, first-order derivative preprocessing and elastic network modelling algorithm was the most effective, and the model had a coefficient of determination of prediction set (R_P²)=0.72, root mean squared error of prediction set (RMSEp)=63.28 mg∙kg^-1, mean absolute error of prediction set (MAEp)=49.03 mg∙kg^-1, and performance deviation ratio (ratio)=1.90. The best K content prediction model was developed based on the CARS feature selection algorithm, raw spectra and support vector machine modelling algorithm, which had R_P²=0.57, RMSEp=141.49 mg∙kg^-1, MAEp=116.24 mg∙kg^-1, RPD=1.57. Mg content prediction model developed based on CARS feature selection algorithm, raw spectra and partial least squares regression modelling algorithm was the most effective, the model R_P²=0.51, RMSEp=12.08 mg∙kg^-1, MAEp=9.84 mg∙kg^-1, and RPD=1.30. 【Conclusion】 It was feasible to use MIRS to predict Na and K content in milk from Chinese Holstein cows, which could predict Na content with a high degree of accuracy and approximate quantitative prediction of K content (for distinguishing between low and high K concentration samples). The use of the CARS algorithm to extract the characteristic bands before modelling improved the accuracy of the MIRS prediction model, and greatly reduced the computing time to improve the efficiency of the MIRS model in predicting phenotypic data.

【Background】 Bovine mastitis is one of the most serious diseases in dairy cows, which seriously affects milk quality and not only causes economic losses, but also even jeopardizes human health. Previous studies have shown that lncRNAs were widely involved in inflammation and immune regulation in humans and animals. lncRNA RRAS2-AS1 was a newly identified differentially expressed lncRNA in our previous study, and its expression pattern and function are still unclear. 【Objective】 The aim of this study was intended to investigate the mechanism of lncRNA RRAS2-AS1 in the inflammatory response of bovine mammary epithelial cells (bMECs) in dairy cows, so as to provide a theoretical basis for the resolution of molecular regulatory mechanisms of bovine mastitis and molecular breeding for anti-mastitis.【Method】 The cloning of lncRNA RRAS2-AS1 was performed by RT-PCR and RACE, and the target genes prediction and functional enrichment analysis of lncRNA RRAS2-AS1 were performed by bioinformatics methods. The bMECs were identified by immunofluorescence staining, and the subcellular localization of lncRNA RRAS2-AS1 was detected by cytoplasmic isolation and semi-quantitative PCR. The expression patterns of lncRNA RRAS2-AS1 in lipopolysaccharide (LPS) induced bMECs inflammatory response and in bovine mammary tissues with mastitis were detected using qRT-PCR. An inflammatory cell model was constructed using LPS-induced bMECs, on this basis, the effects of lncRNA RRAS2-AS1 on the expression of pro-inflammatory cytokine genes, proliferation-related genes and apoptosis-related genes at the mRNA and/or protein levels in inflammatory bMECs were studied using lncRNA overexpression, qRT-PCR and ELISA techniques. At the same time, the effects of lncRNA RRAS2-AS1 on the proliferation, viability, and apoptosis of bMECs were further verified using EdU, CCK-8 and flow cytometry.【Result】 The length of lncRNA RRAS2-AS1 was 363 bp, which was mainly localized in the cytoplasm. The expression assay results showed that, compared with the control group, the expression of lncRNA RRAS2-AS1 was significantly down-regulated in LPS induced bMECs inflammation and in bovine mammary tissues with mastitis (P<0.05). The results of lncRNA RRAS2-AS1 overexpression assay showed that, compared with the control group, the expression of key genes the inflammatory signaling pathway (TLR4 and NF-κB1), pro-inflammatory cytokine genes (IL-1β, IL-6 and IL-8), pro-apoptotic genes (BAD, CASP3, BAX, etc.) were significantly down-regulated (P<0.01) in the lncRNA RRAS2-AS1 overexpression group, while the expression levels of proliferation marker genes (CDK2, CDK4, and PCNA) were significantly up-regulated (P<0.05). In addition, the cell viability of the lncRNA RRAS2-AS1 overexpression group was significantly increased (P<0.05), while the apoptosis rate of bMECs was significantly reduced (P<0.01).【Conclusion】 lncRNA RRAS2-AS1 was significantly down-regulated in LPS induced bMECs inflammation and in bovine mammary tissues with mastitis. Overexpression of lncRNA RRAS2-AS1 decreased the expression of pro-inflammatory cytokines IL-6, IL-8, and IL-1β at mRNA and protein levels, and promoted cell viability and proliferation and inhibited apoptosis, which attenuated the inflammatory response of LPS-induced bMECs. These and the results laid the foundation for analyzing the molecular regulatory network of mastitis in dairy cows.

【Background】 Many previous studies have reported that copy number variation (CNV) is a kind of deletion or duplication with the length of 50 bp-5 Mb, which can affect the expression of genes. It is closely associated with economically important traits of livestock, which is one kind of promising molecular markers. Lion-head goose is one of the largest goose species in the world. It is originated in Raoping, Guangdong Province and is the raw material for Guangdong marinated geese. So far, there has no genome-wide association study on investigating the relationship between CNV and body weight and size in lion-head geese. 【Objective】 This study identified the CNV and CNV region (CNVR) of lion-head geese by using the second-generation genome sequencing data, and then detected CNV and candidate genes significantly affecting body weight and size through the association between them, which could provide the valuable reference information for molecular breeding of lion-head geese. 【Method】 A total of 111 lion-head geese were collected from Baisha Poultry and Livestock Origin Research Institute in Shantou, including 20 males and 91females. All geese were raised and managed under the uniform standards. The body weight and size traits of 111 geese were measured, and the body size traits included body oblique length, chest depth, chest width and so on. The next-generation genome sequencing data (5×) was generated using blood samples for these geese. SOAPnuke was used for the quality control of sequencing data.The BWA module of Speedseq was used for alignment, and the LUMPY and CNVnator modules of Speedseq were used to detect structural variations (SVs). CNV were selected from SV. The software SVtools was used to genotype CNV, and the association analysis between CNV and body weight and size traits was performed by using the single maker mixed model. CNV significantly associated with traits was screened through the chromosome significance level (0.05/number of CNV on the chromosome), and then annotated the significant CNV including their upstream and downstream 50 kb to identify candidate genes for the body weight and size of lion-head geese. The R package CNVrd2 was used to analyze the linkage disequilibrium (LD) of chromosome-significant CNV and chromosome-significant SNP with physical distance less than 1 Mb. 【Result】 For 111 lion-head geese, this study detected 99158 CNV including 94 560 deletions and 4 598 duplications. The average length of CNV was 11 858 bp, and most (74.06%) of them were located in the range of 50 bp-1 Kb. A total of 5 225 CNVR were detected, which contained 5 029 loss types, 110 gain types, and 86 mixed types. The average length of CNVR was 7 136 bp, and the lengths of most (81.03%) of the CNVRs were 50 bp-1 Kb. Functional annotation showed that 46.92% of CNVR were located in the inter gene region, 10.30% were located the upstream, and 9.35% were located the downstream. There were 6 217 CNV accurately genotyped for association analysis. By the association analysis of body weight and size traits and CNV, a total of 55 CNV exceeded the significance level of chromosomes, and then annotated 45 candidate genes based on these 55 CNV. Among these 45 candidate genes, it was found that 10 genes, such as SETD2, UBR7 and G2E3, simultaneously influenced two or more traits. Chromosome-significant CNV affected body weight and size traits independently of chromosome-significant SNP (r²<0.02). 【Conclusion】 This study for the first time reported the distribution of CNV and CNVR in the genome of lion-head geese as well as the association between CNV and body weight and size by using the next-generation genome sequencing data. It was found that a total of 45 candidate genes influencing the body weight and size traits, in which 11 genes were reported to be related to signal pathways of animal growth, among these 11 genes, SETD2, UBR7, ASB1 and HDAC4 were involved in muscle proliferation, differentiation and metabolism, G2E3, P3C2B, NOVA1 and PDE1B were involved in adipogenesis and obesity, ILKAP was involved in regulating growth factors, KIF1B was involved in bone metabolism, and ZFP37 was involved in glycogen metabolism. These results laid a solid foundation for analyzing molecular genetic mechanism and detecting molecular marker for the growth performance of lion-head goose.