Table of Content

01 August 2025, Volume 58 Issue 15

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Identification and Characterization of Retrotransposon Tos17 in the Genomes of Indica Rice

LUO JiaRui, WU SanLing, GUO Fu, LIU Zhen, SONG JingHan, TAN YuanYuan, SHU QingYao

Scientia Agricultura Sinica. 2025, 58(15):  2933-2947.  doi:10.3864/j.issn.0578-1752.2025.15.001

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【Objective】 Tos17 is a type of retrotransposon in the rice genome. In the japonica variety Nipponbare, a Tos17 located on chromosome 7 (Tos17^Chr.7) can be activated during tissue culture. This study aims to reveal the genomic features of Tos17 of indica varieties in China and determine whether their Tos17 can be activated in tissue culture like in japonica rice, which may affect biotechnological breeding. 【Method】 High-quality genome resequencing data of indica varieties or hybrid parents were retrieved from public databases. An in-house program was developed to identify and analyze Tos17 insertion loci, confirmed by IGV visualization and PCR assays. The varieties were classified through hierarchical clustering and principal component analysis, a phylogenetic tree was constructed based on genome-wide single nucleotide polymorphism (SNP) data, and the association between varietal clusters and Tos17 haplotypes was assessed using Mantel test. Transgenic plants were generated by Agrobacterium -mediated transformation of mature embryo-derived callus of indica varieties, and the changes of Tos17 copy number were analyzed in 125 T₀ transgenic plants.【Result】 23 distinct Tos17 insertion loci were identified in 1 511 indica varieties using the Tos17-finder, a program developed specifically for Tos17 identification. All varieties had a Tos17 on Chr.10 (Tos17^Chr.10) identical to the one in japonica rice Nipponbare, and there were two high-frequency Tos17 copies on Chr.2, i.e., Tos17^Chr.2-1 (79.0%) and Tos17^Chr.2-2 (83.7%), but only 85 (5.6%) varieties carried the Tos17^Chr.7 common to japonica rice. There were 4.0 Tos17 copies per variety on average, and while 11 varieties had up to 8 Tos17 copies, 35 only had a single Tos17, i.e., Tos17^Chr.10. Twelve Tos17 insertions were located within or 2 kb up- or down-stream of annotated genes, with the remaining 11 in intergenic regions. Phylogenetic analysis based on SNPs classified the 1 511 varieties into three subpopulations, each showing partial correlation with specific Tos17 haplotypes. No new Tos17 insertions were detected in the 125 T₀ transgenic seedlings of 5 indica varieties. A molecular marker capable of accurately distinguishing Tos17^Chr.7 from other Tos17s was developed. 【Conclusion】 The genomic features of Tos17 in indica rice varieties differ from those in japonica rice variety Nipponbare. The developed molecular marker can be used to determine readily whether the test materials carry the activatable Tos17^Chr.7.

Genetic Diversity Analysis of Protein Fractions and Quality in Xinjiang Winter Wheat Cultivar Resources

LIU PengPeng, LI JiangBo, XU HongJun, NIE YingBin, HAN XinNian, KONG DeZhen, SANG Wei

Scientia Agricultura Sinica. 2025, 58(15):  2948-2959.  doi:10.3864/j.issn.0578-1752.2025.15.002

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【Objective】 To provide high-quality parental materials for broadening the genetic foundation of Xinjiang wheat resources as well as parental selection and cultivar development in breeding programs, the genetic diversity and relationships of protein quality traits and storage protein components in Xinjiang wheat germplasm resources were analyzed in this study. 【Method】 A total of 303 Xinjiang wheat accessions were evaluated for variation, correlation, and cluster analysis of protein quality traits and storage protein component contents. Genetic diversity indices (H') and membership function values were calculated to comprehensively assess the materials.【Result】 The coefficients of variation (CV) for protein quality traits and storage protein component contents in Xinjiang wheat resources ranged from 5.52% to 60.99% and 9.17% to 23.69%, respectively. The highest CV for protein quality traits was observed in 8-minute width (60.99%), while the highest CV for storage protein components was found in unextractable polymeric protein (UPP, 23.69%). Genetic diversity indices ranged from 1.06 to 2.15 with an average of 1.78. In addition, the gluten index (GI) exhibiting the highest value (2.15) and peak time (PT) the lowest (1.06). Correlation and multiple regression analyses demonstrated that the comprehensive evaluation value (F₁₅) effectively assessed protein quality (gluten quality). Key traits for protein quality evaluation included gluten index (GI), peak time (PT), 8 minute width, sedimentation value (SV), and unextractable polymeric protein (UPP), which are applicable for future breeding applications. Cluster analysis classified the 303 accessions into three groups (15.84%, 43.23%, and 40.92%, respectively). Group I exhibited the highest comprehensive evaluation value (F₁₅) and optimal quality indices, with significantly superior means for seven parameters, including percentage of unextractable polymeric protein (%UPP), unextractable polymeric protein (UPP), gluten index (GI), peak time (PT), 8 minute width, 8 minute area, and sedimentation value (SV). This confirmed the reliability of F₁₅ in evaluating gluten quality. 【Conclusion】 The genetic diversity distribution patterns and relationships of protein quality traits and storage protein components in Xinjiang winter wheat resources were elucidated. Critical traits for protein quality evaluation were identified, and a set of accessions with superior comprehensive performance in storage protein components and protein quality traits were selected based on F₁₅. These resources hold significant potential for utilization in future wheat breeding programs of Xinjiang.

Genomic Selection Method Based on G2PSE Stacking Ensemble

ZHUANG RunJie, LIU HuiMing, WANG ShiYu, LÜ WanPing, WEN YongXian

Scientia Agricultura Sinica. 2025, 58(15):  2960-2979.  doi:10.3864/j.issn.0578-1752.2025.15.003

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【Objective】 Genomic selection (GS) is a core technology for predicting individual phenotypes or genetic values from genome-wide marker information, which has important theoretical value and practical significance in agricultural breeding and genetic research. However, high-dimensional feature redundancy and nonlinear relationship modeling are key challenges in genomic selection. A genotype to phenotype stacking ensemble (G2PSE) is proposed, aiming to improve the prediction accuracy and generalization ability, and provide an efficient solution for high-dimensional genomic data analysis. 【Method】 The G2PSE stacking ensemble model framework was constructed, incorporating ten-fold cross-validation, ensemble learning, feature selection (LAR algorithm), and feature enhancement strategies. The model employed random forests (RF), support vector regression (SVR), and gradient boosting regression (GBR) as base learners, with ordinary least squares regression (OLSR) as the meta-learner. Additionally, the impact of meta-learners such as random forest, support vector regression, and neural networks on model performance was evaluated. The G2PSE model consisted of three core submodels: (1) All-feature stacking ensemble (AFSE), which fully utilized all SNP features; (2) LAR-feature stacking ensemble (LFSE), which reduced redundant information through feature selection to improve generalization; (3) LAR-feature enhanced stacking ensemble (LFESE), which combined feature selection with enhancement strategies to optimize prediction capability in high-dimensional data environments. The performance of three feature enhancement variants (AFESE, HFESEⅠ, HFESEⅡ) was explored. Finally, the model was evaluated experimentally on multi-trait datasets of three species, namely wheat, soybean, and tilapia, and further evaluated on an independent test set using the Pepper203 dataset to validate the robustness of the model. 【Result】 The G2PSE model significantly outperformed traditional methods and single machine learning models in two metrics, Pearson correlation coefficient (PCC) and mean absolute error (MAE). Among the three core submodels, LFESE performed the best by combining the feature selection and enhancement strategies, LFSE reduced redundant information and enhanced the generalization ability by feature selection, and AFSE had a significant advantage in comprehensively capturing genotypic global information. In addition, the three feature enhancement variant models further validated the importance of feature quality compared to feature quantity in improving prediction performance. The experiments also showed that the linear regression model performed best in meta-learner selection, while the LFESE and LFSE submodels demonstrated a more balanced performance in terms of computational efficiency. And a reasonable feature selection threshold was crucial for model performance, where the optimal threshold for low-dimensional datasets was 10%-20%, while the optimal threshold for high-dimensional datasets was 1%. Finally, the evaluation on an independent test set proved that the LFESE submodel had the best generalization ability. 【Conclusion】 The G2PSE model significantly improves genomic selection prediction performance through ensemble learning, feature selection, and enhancement strategies.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Relationship Between Boll Morphological Characteristics and Fiber and Kernel Quality of Gossypium hirsutum L. and Gossypium barbadense L.

TANG ChaoYuan, LIU TaoFen, WU YanQin, ZHANG QiPeng, LI ZiLiang, CHEN YunRui, LEI ZhangYing, ZHANG YaLi, ZHANG WangFeng, DU MingWei, YANG MingFeng, TIAN JingShan

Scientia Agricultura Sinica. 2025, 58(15):  2980-2992.  doi:10.3864/j.issn.0578-1752.2025.15.004

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【Objective】 The analysis of the differences and relationships of boll morphological characteristics, boll weight, fiber and kernel quality between G. hirsutum and G. barbadense were carried out in this study, so as to provide a theoretical basis for the breeding of new cotton varieties and the cultivation of high yield and quality. 【Method】 The experiment was carried out in Xinjiang Wulan Wusu Agricultural Meteorological Station from 2022 to 2023. 299 accessions of G. hirsutum and 274 accessions of G. barbadense were selected, and the effects of boll morphological characteristics on fiber and kernel quality of G. hirsutum and G. barbadense were determined by correlation, principal component analysis and grey correlation analysis. 【Result】 The boll length of G. barbadense varied from 3.79 to 6.20 cm, which was significantly higher than that of G. hirsutum, while the boll diameter, surface area and volume of G. hirsutum were higher than those of G. barbadense, and the boll diameter ranged from 2.81 to 4.27 cm, the surface area and volume were 21.86 to 37.42 cm² and 14.76 to 33.58 cm³, respectively; the strength of G. hirsutum increased with the increase of boll diameter, and the fiber quality of G. barbadense increased with the increase of boll volume. The bur weight of G. barbadense was 0.97 g, which was significantly lower than that of G. hirsutum by 37.01%. The bur mass per area of G. hirsutum was significantly higher than that of G. barbadense, and its variation range was 2.24 to 9.88 g·dm^-2. The upper half mean length and strength of G. hirsutum were significantly positively correlated with bur weight and bur mass per area; the upper half mean length of G. barbadense was significantly negatively correlated with bur weight and bur mass per area, and the strength was significantly positively correlated with bur weight. The increase of bur weight and bur mass per area were significantly positively correlated with the increase in boll weight. The fiber weight and seed weight of G. hirsutum ranged from 0.85 to 3.69 g and 1.85 to 6.16 g, which were 48.34% and 37.97% higher than those of G. barbadense, respectively. The upper half mean length and strength of G. hirsutum were significantly positively correlated with seed weight, while G. barbadense was significantly negatively correlated with fiber weight. The oil content of G. barbadense kernel was 35.04%, significantly higher than that of G. hirsutum by 1.75%, and the protein content of G. hirsutum kernel was 40.75%, significantly higher than that of G. barbadense by 2.86%. There was a significant negative correlation between fiber quality and kernel protein content. With the increase of protein content, the upper half mean length and strength decreased significantly. 【Conclusion】 Therefore, the difference in the distribution of photosynthetic products in fiber and kernel was the main reason for the difference in fiber quality between G. hirsutum and G. barbadense. Expanding the storage capacity of cotton boll could synergistically increase the boll weight and strength of G. hirsutum and G. barbadense.

The Physiological Mechanism of Graphene Oxide-Induced Enhancement of Peanut Seed Germination and Seedling Salinity Tolerance

LI QiongWei, BI YanLiang, YAN Ning, ZOU XiaoXia, SI Tong

Scientia Agricultura Sinica. 2025, 58(15):  2993-3006.  doi:10.3864/j.issn.0578-1752.2025.15.005

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【Objective】 As a novel carbon nanomaterial, graphene oxide (GO) is rapidly emerging as a promising tool in modern agriculture. This study aimed to investigate the effects of GO on seed germination and seedling salt tolerance by integrating physiological indices with transcriptomic analyses. The findings would establish a theoretical foundation for the salt tolerance-enhancing effects of GO in peanut cultivation, and provide a scientific basis for its application in improving plant salt tolerance under saline environments. 【Method】 The Q7 peanut variety was used for the experiments, treated with 200 mg·L^-1 GO, and subjected to seed germination and seedling salinity tolerance tests. The changes in seed germination indices, seedling growth characteristics, photosynthetic characteristics, antioxidant characteristics, and root transcriptomics were analyzed to explore the physiological mechanisms by which GO priming enhances peanut salinity tolerance. 【Result】 During the seed germination stage, seeds treated with GO showed higher exposure rate of the white embryo and germination rate. At the seedling stage under normal conditions, compared with the control treatment, GO priming increased root volume, root surface area, chlorophyll a and chlorophyll b content by 12.96%, 12.59%, 52.33% and 25.19%, respectively, while the dry weight of aboveground and underground parts decreased by 6.01% and 17.82% compared with the control treatment, respectively. Under salt stress, GO priming significantly increased plant height, dry weight of aboveground and underground parts, total root length, and root surface area by 30.67%, 20.25%, 27.70%, 18.04% and 17.78%, respectively; total chlorophyll, chlorophyll a content, chlorophyll b content, and maximum photochemical efficiency (Fv/Fm) were significantly increased by 149.39%, 126.27%, 126.27% and 18.26%, respectively, and the net photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr) of leaf gas exchange parameters were increased by 66.69%, 138.99% and 173.39%, respectively. The relative water content of leaves was increased by 83.32%; the activities of antioxidant enzymes SOD and APX in leaves were significantly increased by 59.62% and 104.70%, respectively, while the intercellular CO₂ concentration (Ci), relative electrical conductivity, malondialdehyde (MDA) content, and relative electrical conductivity (REC) were significantly decreased by 19.58%, 14.73%, 56.23% and 72.87%, respectively. Moreover, after GO priming, the differentially expressed genes in peanut seedlings were mainly involved in plant secondary metabolites, photosynthesis, and carbon and nitrogen metabolisms. 【Conclusion】 GO priming enhanced seed germination, and under salt stress, GO priming promoted the growth of peanut seedlings, including higher plant height, root length, and plant biomass. In addition, GO priming mediated the photoprotection of photosynthetic mechanisms, which was manifested as higher Pn, Fv/Fm, and total chlorophyll content under salt stress. Meanwhile, the activities of antioxidant enzymes in GO-primed peanut leaves were significantly increased, thereby reducing salt-induced MDA content and REC to maintain the integrity of the plasma membrane. GO played a unique role in promoting seed germination, alleviating salt stress, and increasing peanut pod yield by regulating multiple physiological processes.

The Impact of Nitrogen Addition on the Species Stability of Artificial Grasslands Under Different Planting Patterns

YUAN Bo, XU LiJun, CUO MeJi, NIE YingYing, ZHANG HongZhi, LIU XinWei, YANG Min, GUO MingYing

Scientia Agricultura Sinica. 2025, 58(15):  3007-3019.  doi:10.3864/j.issn.0578-1752.2025.15.006

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【Objective】 The agro-pastoral ecotone in northern China is an important production base for agricultural and livestock products and a crucial ecological security barrier. This study analyzed the effects of different planting patterns on pasture yield, interspecific compatibility, and yield stability, aiming to provide the important scientific evidence for revealing the sustainable utilization of artificially established grasslands under different planting patterns with nitrogen addition.【Method】 Based on the long-term cultivated grassland core experimental platform at the Hulunbuir National Field Station, three planting patterns were established: single sowing of Medicago varia (M), single sowing of Bromus inermis (B), and a 1:1 mixed sowing of Medicago varia and Bromus inermis (M:B). Additionally, three nitrogen application levels were set: no nitrogen (CK), 75 kg N·hm^-2 (N₇₅), and 150 kg N·hm^-2 (N₁₅₀). Indicators, such as yield, relative yield (RY), relative yield total (RYT), competition ratio (CR), stability, and asynchrony, were utilized to investigate the response of forage yield to nitrogen addition under different planting patterns in artificial grasslands.【Result】 (1) Through eight years of long-term site-specific observations, the results showed that both N₇₅ and N₁₅₀ treatments significantly increased the forage yield of mixed-sown grasslands in the early stages. Specifically, the N₁₅₀ treatment promoted the forage yield of monoculture grasslands. Moreover, under different nitrogen application levels, the forage yield and stability of mixed-sown grasslands were consistently higher than those of monoculture grasslands. (2) In the mixed-sown grassland of variegated alfalfa and smooth brome, the competition ratio of variegated alfalfa was greater than 1 in the first to fifth year after establishment, and less than 1 in the sixth to eighth year. This indicated that variegated alfalfa was highly competitive in the early stages, while smooth brome maintained system balance through continuous nutrient utilization in the later stages. This temporal complementarity enhanced the forage yield stability of the mixed-sown grassland by 1.41-1.53 times and the asynchrony by 1.27-1.57 times, thereby buffering yield fluctuations. Furthermore, there was a positive correlation between stability and asynchrony.【Conclusion】 Species yield asynchrony and stability were key factors determining the stability of mixed-sowing yields. For establishing monoculture artificial grasslands of variegated alfalfa or smooth brome in the Hulunbuir region, it was recommended to apply 150 kg N·hm^-2 of nitrogen. For establishing mixed-sown grasslands of variegated alfalfa and smooth brome, it was advisable to increase the nitrogen application rate to 75 kg N·hm^-2 during the initial establishment phase. Through complementary symbiosis, legume-grass mixed-sown pastures could effectively utilize soil nutrients, increase forage yield, and simultaneously reduce the amount of exogenous nitrogen applied, thereby achieving the goal of promoting and maintaining high pasture productivity.

PLANT PROTECTION

Function of Effector Pg00778 Regulation on the Pathogenicity of Pyrenophora graminea to Barley

YANG WenJuan, GAO JiaCheng, WANG YanTing, LI Yan, GUO Ming, WANG JunCheng, MENG YaXiong, WANG HuaJun, SI ErJing

Scientia Agricultura Sinica. 2025, 58(15):  3020-3035.  doi:10.3864/j.issn.0578-1752.2025.15.007

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【Objective】 Barley leaf stripe is a significant agricultural disease caused by Pyrenophora graminea, which severely affects the yield and quality of barley. During the process of infecting its host, P. graminea secretes a variety of effector proteins to modulate the plant’s defense system, thereby enhancing its pathogenicity. The objective of this study is to clarify the function and mechanism of action of the P. graminea effector protein Pg00778, and to provide a theoretical reference for the study of P. graminea.【Method】 Based on the previously sequenced genome data of the highly virulent P. graminea strain QWC from our research group, combined with transcriptomic analysis of the P. graminea-highly susceptible barley interaction, a candidate effector protein, Pg00778, was identified. Using the P. graminea wild-type strain QWC as the experimental material, the Pg00778 was cloned, sequence characteristics were analyzed, and a phylogenetic tree was constructed using MEGA11.0 with homologous proteins from P. graminea and other pathogenic fungi. The virulence function of Pg00778 was validated through transient expression in Nicotiana benthamiana leaf cells, while its subcellular localization was determined using tobacco leaf localization assays. Both RNAi mutants and overexpression transformants of Pg00778 were generated via CaCl₂-PEG4000-mediated protoplast transformation. Subsequent assessments of mycelial growth and pathogenicity were conducted to elucidate the function of Pg00778 during P. graminea infection in barley.【Result】 Pg00778 lacks signal peptides, transmembrane domains, and known functional domains, categorizing it as a non-classical effector protein. Phylogenetic analysis indicated that Pg00778 has the closest evolutionary relationship with the P. tritici-repentis (XM_066107220.1). The transient expression of Pg00778 in N. benthamian leaves was ineffective in inducing cell death and inhibiting cell death caused by BAX. Pg00778 is mainly localized in the nucleus, cell membrane and cytoplasm. Compared with the wild-type strain QWC, the relative gene expression levels of the Pg00778 in RNAi mutants were reduced by 41.6% and 54.0%, respectively, while the relative gene expression levels of the Pg00778 in OE mutants increased by 290.0%, 397.4%, and 263.7%, respectively. A significant reduction was detected in the growth rate of the RNAi mutants by 21.4% to 30.1%, and an increase in the relative content of chlorophyll in barley leaf by 34.1% to 39.1% after infection. The colony growth rate of the Pg00778-OE mutants was significantly faster than that of the wild-type strain by 7.3% to 12.6%, and the relative content of chlorophyll in barley leaf infected by the OE mutants was significantly lower than that of the wild-type strain by 20.0% to 23.0%. There was no significant difference in the microscopic morphology of mycelium. Pathogenicity test revealed that the incidence of Pg00778-RNAi mutants decreased by 51.5% and 49.0% compared to the QWC, while the Pg00778-OE mutants increased by 19.0%, 20.3%, and 21.2%. Compared with ‘Alexis’ infected by wild-type strain QWC, ‘Alexis’ infected by Pg00778-OE mutants showed more severe stripe disease symptoms, deeper and bluer trypan blue staining, and significantly increased accumulation of reactive oxygen species in the leaves; the stripe invasion symptoms of leaf infected by Pg00778-RNAi mutants were significantly reduced, the cell membrane was more intact, and the accumulation of reactive oxygen species was significantly reduced.【Conclusion】 Effector protein Pg00778 positively regulates the pathogenicity of P. graminea and serves as a crucial effector protein influencing its infection to the host.

SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT

Effect of Applying Selenium Fertilizer to Soil on Selenium Enrichment of Winter Wheat in the Different Wheat-Producing Areas of China

LI HaiFeng, LI WenHu, LI YuKe, WANG ZhaoHui, LIU JinShan

Scientia Agricultura Sinica. 2025, 58(15):  3036-3050.  doi:10.3864/j.issn.0578-1752.2025.15.008

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【Objective】 Based on varying climate, soil, tillage, and field management conditions in different wheat-producing regions of China, this study aimed to investigate the impacts of soil-applied selenium (Se) fertilizer on Se enrichment in wheat and the available Se content in soil. Furthermore, the regional factors contributing to differences in the enhancement of Se content in wheat grain across these diverse areas were analyzed. These findings would provide a foundation for the efficient utilization of Se fertilizer and the scientific advancement of biofortified Se in wheat grain. 【Method】 The experiment was conducted from 2022 to 2024 in four representative wheat-producing regions of China, including Yongshou County of Shaanxi Province, Baixiang County of Hebei Province, Zitong County of Sichuan Province, and Shucheng County of Anhui Province. Five different levels of selenite fertilizer were applied: 0 (Se0), 100 (Se100), 200 (Se200), 300 (Se300), and 400 (Se400) g·hm^-2. At the flowering and maturity stages of wheat, wheat plant and soil samples in each plot were collected. The wheat biomass, grain yield, Se content of different wheat organs, Se uptake and soil available Se content in different wheat-producing regions were determined and analyzed. 【Result】 Soil-applied Se fertilizer had no significant effect on wheat yield and aboveground biomass. Selenium content and accumulation in wheat organs were significantly increased with the increase of Se application rate. There was a linear positive correlation between Se content and Se application rate, and the selenium enrichment effect was in the order of Baixiang of Hebei Province > Yongshou of Shaanxi Province > Shucheng of Anhui Province > Zitong of Sichuan Province, and the Se content of grain was increased by 1.03, 0.57, 0.35, and 0.33 μg·kg^-1, respectively with each application of 1 g·hm^-2 selenium. Under the same Se application level, TF_{spike/stem and leaf} value at flowering stage was higher than TF_{stem and leaf/root} value, and TF_{grain/stem and leaf} value at maturity stage was increased with the increase of Se application rate, indicating that selenite was not easily transferred from root to stem and leaf, but easily transferred from stem and leaf to ear. The soil available Se content was significantly increased with the increase of Se application rate and the increasing effect was in the order of Yongshou of Shaanxi Province > Baixiang of Hebei Province > Shucheng of Anhui Province > Zitong of Sichuan Province, and the soil available Se content at the four sites was increased by 1.141, 0.077, 0.008, and 0.008 μg·kg^-1, respectively with a rate of 1 g·hm^-2 selenium application. The Se fertilizer application rate to meet the enrichment of grain selenium (150 μg·kg^-1) in Yongshou of Shaanxi Province, Baixiang of Hebei Province, Zitong of Sichuan Province and Shucheng of Anhui Province were 232, 0, 376, and 354 g·hm^-2, respectively. 【Conclusion】 In the wheat fields with low soil Se content, the application of Se fertilizer could lead to the production of Se-enriched wheat grain and an increase in the soil Se content in various wheat-producing regions of China. However, in alkaline soils, the enrichment of Se in wheat grain was more effective and the enhancement of soil available Se content was greater. The input rate of Se fertilizer for selenium enrichment requirement of wheat (>150 μg·kg^-1) was lower in the alkaline soil than that in the acid soils.

Conservation Tillage and Nitrogen Application Promote Soil Carbon and Nitrogen Mineralization and Improve Maize Photosynthetic Characteristics and Yield

LU YiNing, GU XiaoBo, DU YaDan, LI XiaoYan, YAN TingLin, ZHAO TongTong

Scientia Agricultura Sinica. 2025, 58(15):  3051-3063.  doi:10.3864/j.issn.0578-1752.2025.15.009

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【Objective】 Exploring the effects of conservation tillage and nitrogen application on soil carbon and nitrogen mineralization characteristics, as well as their impact mechanisms on maize photosynthetic characteristics and yield, in order to provide a basis for improving soil fertility and promoting maize production in arid areas of Northwest China.【Method】 This study conducted a two-year (2019-2020) maize field location experiment, with traditional tillage (CT) as the control, and set up three protective tillage measures (no tillage: NT; no tillage in wheat season and rotary tillage in maize season: OT; ridge cultivation with no tillage: RNT) and two nitrogen application levels (N0: 0; N2: 170 kg N·hm^-2), for a total of 6 treatments. The effects of conservation tillage and nitrogen application on soil nutrient content, carbon and nitrogen mineralization characteristics, maize photosynthetic physiological characteristics, and yield were investigated systematically. 【Result】 Conservation tillage and nitrogen application significantly increased soil nutrient content (P<0.05). Under N2 level, compared with CT treatment, NT, OT, and RNT treatments increased soil organic carbon (SOC), microbial biomass carbon (MBC), total nitrogen (TN), and microbial biomass nitrogen (MBN) content by 8.6%-24.7%, 18.9%-27.0%, 8.9%-20.2%, and 0.3%-24.9%, respectively. The application of nitrogen significantly increased the accumulation of soil carbon mineralization (C_min), nitrogen mineralization accumulation (N_min), and their mineralization rates. Conservation tillage further improved the soil carbon and nitrogen mineralization characteristics. C_min and N_min reached their maximum values under RNTN2 and OTN2 treatments, respectively, which increased by 4.0%-30.2% and 8.0%-52.4% compared with other treatments. Conservation tillage and nitrogen application significantly increased the net photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs) of maize leaves (P<0.05). The Pn, Tr, and Gs of maize reached their maximum values in the OTN2 treatment for two years. The yield of maize in two years showed the order of OTN2>RNTN2>NTN2>CTN2>NTN0>CTN0, with the highest yields of 10.52 and 10.91 t·hm^-2, respectively, which increased by an average of 24.5% and 27.5% compared with other treatments. Based on structural equation modeling analysis, it was found that conservation tillage mainly increased soil nutrient content, promoted soil organic carbon and nitrogen mineralization, enhanced soil available nitrogen supply capacity, and thus promoted the enhancement of maize photosynthetic capacity, achieving maize yield increase. 【Conclusion】 In the arid northwest region, conservation tillage and nitrogen application were of great significance in promoting soil carbon and nitrogen mineralization, increasing maize yield, and maintaining soil productivity. It was recommended that no tillage in wheat season and rotary tillage in maize season combined with nitrogen application was the optimal management measures for increasing maize yield and efficiency.

HORTICULTURE

Effects of Streptomyces sp. TOR3209 on Chlorophyll Synthesis and Polyamine Content in Tomato Plants Under Low Temperature Stress

MA Jia, PENG JieLi, WANG Xu, JIA Nan, LI MengKai, HU Dong

Scientia Agricultura Sinica. 2025, 58(15):  3064-3080.  doi:10.3864/j.issn.0578-1752.2025.15.010

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【Objective】 This study aims to elucidate the biological mechanisms underlying the mitigation of low temperature-induced damage in tomato plants by Streptomyces sp. TOR3209, with particular emphasis on evaluating the impact of TOR3209 inoculation on light-harvesting complex of PSII (LHCII). The investigation focuses primarily on the alterations in chlorophyll biosynthesis pathways and polyamine accumulation.【Method】 Tomato seedlings at the quadrifoliate stage were inoculated with Streptomyces sp. TOR3209 during transplantation into the pots. Low-temperature stress (5 ℃) was conducted 30 days post-transplantation. The experimental design comprised four treatment groups: TOR3209-inoculated plants (TOR3209), non-inoculated plants (NI), TOR3209-inoculated plants exposed to cold stress (TOR3209+C), and non-inoculated plants exposed to cold stress (NI+C). The conformational alterations in LHCII were analyzed through sucrose density gradient centrifugation. Chlorophyll content, precursor metabolite levels, and enzymatic activities in chlorophyll biosynthesis pathways were quantified using microplate-based enzymatic assays. Polyamine speciation profiles were determined by high performance liquid chromatography, while the expression differences of key genes involved in chlorophyll and polyamine biosynthesis metabolism were examined via quantitative real-time PCR.【Result】 Low temperature stress markedly decreased the abundance of LHCII monomers and trimers in thylakoid membranes, while TOR3209 inoculation effectively alleviated the degradation. The chlorophyll content in tomato leaves showed a pronounced decrease by low temperature, concurrent with substantial accumulation of the chlorophyll precursors porphobilinogen (PBG). This accumulation coincided with a marked decline in the level of downstream chlorophyll precursors, including uroporphyrinogen III (Urogen III), protoporphyrin IX (Proto IX), Mg-protoporphyrin IX (Mg-proto IX), and protochlorophyll (Pchl). The reduction in chlorophyll content was mainly caused by the blocked site, which was from PBG to Urogen III in chlorophyll synthesis. Streptomyces sp. TOR3209 enhanced porphobilinogen deaminase (PBGD) enzymatic activity and upregulated expression of the encoded gene HemC. This regulatory effect mitigated the inhibitory impact of low temperature stress on the metabolic conversion from PBG to Urogen III, consequently facilitating the recovery of the previously impaired chlorophyll biosynthesis pathway. Concurrently, the inoculation suppressed both chlorophyllase (Chlase) activity and Clh gene expression levels, promoting chlorophyll accumulation. The contents of free and conjugated polyamines were all decreased significantly under low temperature stress, accompanied by a decreased putrescine (Put)/spermine (Spm) ratio. TOR3209 inoculation alleviated the decline of polyamines, and the levels of different forms of polyamines were significantly elevated at the early stage of stress. Extended stress duration activated the metabolic conversion from Put to spermidine (Spd) through Odc gene upregulation, resulting in progressive accumulation of free/conjugated Put and conjugated Spd. Notably, while Spm levels showed no significant variation relative to NI+C, the elevated ratio of Put/Spm contributed to enhancing the abundance of LHCII and stabilized the aggregation state of LHCII. In addition, TOR3209 effectively reversed the cold stress-induced reduction in transglutaminases (TGase) activity while stimulating polyamine conjugation processes.【Conclusion】 TOR3209 treatment enhanced chlorophyll biosynthesis in tomato leaves through relieving the hindrance on the transformation from PBG to Urogen III caused by low temperature stress, and elevating Put accumulation and the Put/Spm ratio. These biochemical modifications collectively stabilized monomeric and trimeric LHCII configurations, providing comprehensive protection for the photosynthetic apparatus against low temperature stress.

SnRK2.6 Regulates Pollen Tube Growth and ABA Response in Distant Hybridization in Paeonia lactiflora

ZHOU PingXi, WANG JingKun, YOU XiaoLong, HUA Chao, GUO HaoNan, ZHANG MingXing, LIU YiPing, HE Dan, HE SongLin

Scientia Agricultura Sinica. 2025, 58(15):  3081-3096.  doi:10.3864/j.issn.0578-1752.2025.15.011

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【Objective】 This study aims to identify SnRK2 genes from the Paeonia transcriptome to investigate the molecular mechanism of SnRK2 mediates the ABA signaling pathway in regulating pollen germination and pollen tube growth during distant hybridization of Paeonia lactiflora (P. lactiflora). The goal is to provide a theoretical basis for overcoming hybridization incompatibility in P. lactiflora.【Method】 Based on the transcriptome data of P. lactiflora stigmas (NCBI accession number: PRJNA592882), ABA signaling pathway genes and group Ⅲ SnRK2 genes (PlSnRK2.6) were screened and identified. The PlSnRK2.6 gene was cloned and analyzed for bioinformatics analysis and subcellular localization analysis. ABA content in stigmas at 4, 8, 12, 24, and 36 hours post-pollination (self- and cross-pollination) was measured using enzyme-linked immunosorbent assay (ELISA), along with relative gene expression using quantitative real-time PCR (qPCR). The relative expression levels of PlSnRK2.6 in stigmas under exogenous ABA and its inhibitor treatments were analyzed, and yeast two-hybrid (Y2H) assays were conducted to identify PlSnRK2.6-interacting proteins.【Result】 In vitro pollen culture revealed that pollen germination rates and pollen tube lengths were significantly reduced in ABA-supplemented media compared to normal media, indicating that ABA inhibits pollen germination and pollen tube growth in P. lactiflora. Transcriptome analysis identified key genes in the ABA signaling pathway, including KAT1 (ion transport), RBOHF (ROS production), SnRK2 (protein kinase), PP2C (protein phosphatase), ABFs, and ABI family genes. qPCR analysis showed that, compared to self-pollination, the expression of PlSnRK2.6 and PlRBOHF was significantly upregulated, while PlKAT1 expression was downregulated. PlABF2 and PlABI5 exhibited a rise-then-fall expression trend, consistent with transcriptome FPKM trends. The cloned PlSnRK2.6 CDS was 1 092 bp in length, encoding a protein of 363 amino acids. Gene structure analysis revealed a conserved STKc_SnRK2-3 domain. Phylogenetic analysis showed that the SnRK2 family is divided into three subgroups, with PlSnRK2.6 clustering into group Ⅲ alongside SnRK2.6 from Paeonia suffruticosa and Arabidopsis thaliana. PlSnRK2.6 was most closely related to SnRK2.6 in Potentilla indica, with a highly conserved SnRK2-3 domain. Subcellular localization indicated that PlSnRK2.6 is localized in the nucleus. The ABA content in hybrid stigmas was generally higher than in self-pollination. Similarly, PlSnRK2.6 expression was significantly higher during hybridization, particularly at 24 and 36 hours post-pollination, with expression levels 8.94- and 5.07-fold higher than in self-pollination, respectively. ABA responsiveness assays confirmed that PlSnRK2.6 is strongly responsive to ABA, with a more pronounced expression trend in the late stages of pollination. Yeast two-hybrid assays showed that PlSnRK2.6 lacks self-activation activity and interacts with PlACP1 and PlHMGB3.【Conclusion】 An ABA-induced protein kinase, PlSnRK2.6, was identified to be highly expressed in the stigmas of P. lactiflora during distant hybridization and interacts with PlACP1 and PlHMGB3. It is proposed that PlSnRK2.6 and its interacting proteins regulate pollen germination and pollen tube growth via the ABA signaling pathway, thereby playing a key role in modulating hybrid incompatibility in P. lactiflora.

FOOD SCIENCE AND ENGINEERING

The Potential of Alternative Proteins from Edible Fungi Based on Amino Acid and Physicochemical Characterization

YU YunYan, MA GaoXing, DUAN YaNing, TAO Qi, LI XinYi, HU QiuHui, MA Ning

Scientia Agricultura Sinica. 2025, 58(15):  3097-3117.  doi:10.3864/j.issn.0578-1752.2025.15.012

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【Objective】 From the perspectives of nutrition and processing, the amino acid composition, physicochemical properties, and structural characteristics of common edible fungal proteins were analyzed to elucidate the nutritional benefits of proteins derived from fungi, and to explore their prospective applications within the realm of food processing.【Method】 From the vantage points of raw material accessibility and economic advantages, eight species of edible fungi were chosen as subjects for this study: Lentinula edodes, Agaricus bisporus, Hypsizigus marmoreus (white cultivar), Pleurotus ostreatus, Hypsizygus marmoreus (brown cultivar), Agrocybe aegerita, Flammulina filiformis, and Pleurotus eryngii. To evaluate the disparities in amino acid composition and digestibility of their proteins, by utilizing metrics such as ratio of amino acid and amino acid ratio coefficient, the nutritional value and potential benefits of these fungal proteins as substitutes for conventional proteins were assessed by comparing them with whole egg protein, wheat protein, and soybean protein. The physicochemical properties of various edible fungal proteins were analyzed, including solubility, water-holding property, oil-holding property, emulsifying property, and foaming property. The structural characteristics were analyzed by Fourier-transform infrared spectroscopy and scanning electron microscopy. The correlation between physicochemical properties and structural characteristics were established, and the potential applications of edible fungal proteins within the food industry were assessed.【Result】 The EAA/TAA ratios of all eight edible fungal proteins were approximately 40%, while the EAA/NEAA ratios exceeded 60%, aligning with the ideal protein model. The amino acid profile of L. edodes protein exhibited the greatest similarity to the amino acid pattern spectrum and the reference protein's amino acid composition, boasting an essential amino acid index of 66.31. Upon adjusting the amino acid score to account for digestibility, L. edodes protein continued to hold the highest overall score, establishing it as a high-quality protein source. Furthermore, the essential amino acids of A. aegerita protein constituted 47.12% of its total amino acid profile, a figure comparable to that of whey protein, thereby rendering it an exceptional supplementary source of these vital essential amino acids. The digestibility of the protein from H. marmoreus (white cultivar) was 89.59%, and as a protein that is rapidly digestible, it would be suitable for facilitating swift physical recovery post-exercise. The physicochemical properties of the eight edible fungal proteins also exhibited considerable differences. Both H. marmoreus (white cultivar) and H. marmoreus (brown cultivar) displayed favorable water-holding, oil-holding and emulsifying properties, suggesting extensive potential applications within the realm of food processing.【Conclusion】 Utilizing the nutritional evaluation system for edible fungi, the amino acid profile of L. edodes protein has been determined to possess the highest overall rating, thus establishing L. edodes protein as a viable standalone protein supplement. The proteins of P. ostreatus, P. eryngii, H. marmoreus (white cultivar) and H. marmoreus (brown cultivar) possessed processing advantages and structural similarities in various aspects, rendering them suitable for application in the production and processing of meat products, flour-based products, and baked goods, thereby optimizing the utilization of their nutritional and processing benefits.

Effects of Application of the Soy Sauce Residual Squeezed Oil in Diet of Fatten Pigs on Qualities of Pork Meat

TANG JiaLing, ZHANG YuLin, YANG Yong, FANG ZhengFeng, HAN GuoQuan, HUI Teng

Scientia Agricultura Sinica. 2025, 58(15):  3118-3133.  doi:10.3864/j.issn.0578-1752.2025.15.013

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【Objective】 This study aimed to explore the effects of soy sauce residual squeezed oil adding in diet to replace soybean oil on the nutritional value and edible quality of raw pork.【Method】 A total of Duroc × Landrace × large 12 early-weaned piglets of the same age and similar initial weight were selected for unified fattening, which were fed with commercial feed between 25-100 kg. When pig weigh arrive 100 kg, they are randomly divided into 2 groups, 6 replicates in each group, fattening with formula feed; soybean oil was used as the base oil in the diet of the control group, while the experimental group added soy sauce residue pressed oil as the base oil in the base diet, and the additive amount was 2 kg (oil)/100 kg (diet). The test lasted for 52 days until the pig reached 140 kg, which was slaughtered. Following a 24-hour period of acid excretion, the pig's right longissimus dorsi muscle was harvested, and its fat and connective tissue were removed, which were sent to the laboratory via cold chain for analysis of the two groups of raw meat in terms of pH, moisture, protein, fat, lipid composition, reducing sugar, amino acids, color, and electronic nose, among other indicators. 【Result】 In terms of nutritional qualities, the application of oils from soy sauce residual did not affect the content of moisture, protein, fat, 16 amino acids, neutral fat, phospholipids and glucose significantly (P>0.05), but improved the pH of raw meat, while reduced the content of ash, ribose, phospholipid polyunsaturated fatty acids and free fatty acids by approximately 24%, 17%, 30% and 38%, respectively, and the content of saturated fatty acids and n-3 PUFA in neutral lipids were increased by 0.40 and 1.76 times. In terms of eating qualities, the application of oils from soy sauce residual had no significant effect on the texture characteristics, the value of a^* and b^*, however, the L^* value was decreased by 8%, the response of electronic nose changed from alcohols to aldehydes similar compounds, and its water holding capacity had increased.【Conclusion】 The experimental group, it was shown that the soy sauce residual squeezed oil had a certain effect on improving the quality of pork.

ANIMAL SCIENCE·VETERINARY SCIENCE

Expression Pattern of gga-miR-30a-5p and Its Regulation of Abdominal Fat and Intramuscular Fat Deposition in Chicken

HUANG HuaYun, LIU Xing, WANG QianBao, LI RuiRui, YANG MiaoMiao, LI ChunMiao, WU ZhaoLin, KONG LingLin, ZHAO ZhenHua

Scientia Agricultura Sinica. 2025, 58(15):  3134-3144.  doi:10.3864/j.issn.0578-1752.2025.15.014

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【Objective】 miR-30a-5p is a member of the miR-30 family, and its role in chicken fat deposition has not been reported. This study focused on the role of gga-miR-30a-5p in chicken abdominal fat and intramuscular fat deposition, which could lay a foundation for the analysis of the mechanism of chicken abdominal fat and intramuscular fat deposition. 【Method】 This study used high-quality yellow-feathered broilers (dwarf line, S3) and recessive white-feathered chickens (RR) as experimental subjects. Quantitative real-time PCR (qPCR) was employed to detect the expression changes of gga-miR-30a-5p in abdominal fat, liver, and leg muscle tissues at 0, 2, 4, 8, 14, and 16 weeks (0, 2, 4, 8, 14, and 16W), respectively, as well as during the proliferation and differentiation stages of abdominal and intramuscular adipocytes. Abdominal and intramuscular adipocytes were transfected with gga-miR-30a-5p mimics and inhibitors, and qPCR was used to measure the expression changes of gga-miR-30a-5p. Oil Red O staining and isopropanol extraction were performed to assess lipid droplet deposition after transfection. Bioinformatics analysis was conducted to predict the target genes of gga-miR-30a-5p. 【Result】 The expression of gga-miR-30a-5p in different tissues (abdominal fat, liver, and leg muscle) showed significant breed-specific differences (P<0.05). In abdominal fat tissue, the expression of gga-miR-30a-5p in recessive white-feathered chickens at 0W was significantly lower than that in S3 chickens (P<0.05). In both S3 and RR chickens, gga-miR-30a-5p expression in abdominal fat was the highest at 0W, significantly higher than that at other weeks (P<0.05), with no significant differences among other weeks (P>0.05). In liver tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in liver tissue at 16W was significantly higher than that at other weeks (P<0.05), while in RR chickens, its expression at 16W was significantly higher than that at 0 and 8W (P<0.05). In leg muscle tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in leg muscle was the lowest at 14W, significantly lower than that at 2, 8, and 16W (P<0.05), whereas no significant differences were observed among weeks in RR chickens (P>0.05). In abdominal adipocytes, gga-miR-30a-5p expression during the proliferation stage was significantly lower than that at differentiation days 4 and 6 (P<0.05). In intramuscular adipocytes, no significant difference was observed between the proliferation stage and differentiation day 1 (P>0.05), but expression at differentiation days 4 and 6 was significantly higher than during proliferation (P<0.05), gradually increasing with prolonged differentiation. After transfection with gga-miR-30a-5p mimics or inhibitors for 24 hours, gga-miR-30a-5p expression in abdominal adipocytes significantly increased or decreased, confirming successful transfection. Transfection with gga-miR-30a-5p mimics for 3 days significantly enhanced lipid droplet deposition in abdominal adipocytes, whereas transfection with inhibitors significantly reduced deposition (P<0.05). Similarly, in intramuscular adipocytes, transfection with mimics or inhibitors for 24 hours significantly altered gga-miR-30a-5p expression. Mimics transfection for 3 days significantly increased lipid droplet deposition, while inhibitor transfection significantly decreased it (P<0.05). Target gene prediction via GO, Pathway, and protein-protein interaction analyses suggested that UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 were potential target genes of gga-miR-30a-5p in regulating fat deposition.【Conclusion】 gga-miR-30a-5p exhibited significant breed-specific expression differences across tissues, which promoted abdominal and intramuscular fat deposition, with UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 identified as key candidate target genes.

Genome-Wide Association Study of Egg Production Traits in Jinding Duck

LIU HongXiang, ZHANG XuePing, WANG YiFei, WANG ZhiCheng, GU HaoTian, SONG WeiTao, TAO ZhiYun, XU WenJuan, ZHANG ShuangJie, LU LiZhi, LI HuiFang, ZHU ChunHong

Scientia Agricultura Sinica. 2025, 58(15):  3145-3158.  doi:10.3864/j.issn.0578-1752.2025.15.015

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【Objective】 This study aimed to screen and identify single nucleotide polymorphisms (SNPs) and candidate genes associated with egg production traits in Jinding ducks using genome-wide association study (GWAS) technology, providing the reference information for molecular breeding of Jinding ducks. 【Method】 A group of 441 healthy female Jinding ducks from the same batch was used as the research subjects, and the number of eggs laid by each duck from the first egg until 43, 55, and 72 weeks of age (referred to W43, W55 and W72, respectively) was collected. Blood samples were collected from the wing veins of all 441 individuals for whole-genome resequencing. The sequencing data was aligned with the duck reference genome (ZJU1.0) to identify SNPs across the genome. After quality control using plink software to obtain high-quality SNPs, the heritability of egg production at 43, 55 and 72 weeks of age as well as the genetic and phenotypic correlations between each pair of traits were estimated using multi-trait animal model of the ASReml-R 4.2 software. Gemma software was used to separately conduct genome-wide association analyses for the three egg production traits, identifying the associated marker loci shared by them. Then, Bedtools software was employed to annotate these loci and mine for candidate genes near the markers. 【Result】 The heritability of egg production at 43, 55, and 72 weeks of age in Jinding ducks was relatively low, ranging from 0.17 to 0.30, and decreased gradually with increasing age. There was a high genetic positive correlation between egg production at different ages, with higher genetic correlation coefficients between adjacent time points than between non-adjacent time points. The genetic correlation coefficient between 43 and 55 weeks of age was 0.75, and between 55 and 72 weeks of age was 0.89. The GWAS results showed that 174 suggestively significant SNPs (P<9.43×10^-7) were identified for the egg production trait at 43 weeks of age, distributed on chromosomes 3, 13, and 21; 297 suggestively significant SNPs (P<9.43×10^-7) were identified for the egg production trait at 55 weeks of age, distributed on chromosomes 3 and 13; 36 suggestively significant SNPs (P<9.43×10^-7) were identified for the egg production trait at 72 weeks of age, all located on chromosome 3. 20 overlapping suggestively significant SNPs (P<9.43×10^-7) were shared among the three egg production traits, all located on chromosome 3. These loci formed two large haplotype blocks and involved 5 candidate genes, including the VSNL1 gene involved in calcium ion action, the MSGN1 gene involved in mesoderm formation, the KCNS3 gene with potassium ion channel function, and the SMC6 and GEN1 genes involved in DNA repair.【Conclusion】 The genetic parameters of egg production traits at 43, 55 and 72 weeks of age in Jinding ducks were estimated, and 20 suggestively significant SNPs and 5 candidate genes affecting egg production traits at 3 different ages were identified through GWAS, providing the reference information for molecular breeding of egg production traits in Jinding ducks.