Scientia Agricultura Sinica

Cloning and Heat Tolerance Function of Wheat TaGRAS34-5A Gene

DIAO DengChao, LI YunLi, MENG XiangYu, JI SongHan, SUN YuChen, MA XueHong, LI Jie, FENG YongJia, LI ChunLian, WU JianHui, ZENG QingDong, HAN DeJun, $\boxed{\hbox{WANG ChangFa}}$, ZHENG WeiJun

Scientia Agricultura Sinica. 2025, 58(4): 617-634. doi:10.3864/j.issn.0578-1752.2025.04.001

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【Objective】The GRAS family constitutes a unique class of plant-specific transcription factors that play a pivotal role in plant development and stress response. To elucidate the function of GRAS family genes in wheat heat tolerance，which can provide genetic resources and theoretical foundation for wheat heat-resistant breeding.【Method】A potential heat stress-responsive transcription factor gene, TaGRAS34-5A, was identified through transcriptome analysis of TAM107 and Chinese spring wheat seedlings under high-temperature conditions. Subsequently, a bioinformatics analysis was performed on TaGRAS34-5A, and a phylogenetic tree was constructed to elucidate its molecular characteristics. The expression pattern of TaGRAS34-5A under various stresses, including high temperature, abscisic acid (ABA), ethylene (ETH), and salicylic acid (SA) treatments, were examined using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) method. The subcellular localization of the TaGRAS34-5A protein was determined using wheat protoplast transient expression technique. Furthermore, the heat tolerance function of TaGRAS34-5A was validated using the heterologous expression system of Saccharomyces cerevisiae and the BSMV:VIGS (Barley stripe mosaic virus: Virus-Induced Gene Silencing) silencing technique. potential interacting proteins of TaGRAS34-5A were screened using yeast two-hybrid technology, and the heat tolerance function was verified, providing preliminary insights into its heat tolerance mechanism.【Result】TaGRAS34-5A, equipped with a characteristic GRAS domain and belongs to the GRAS transcription factor family, is localized to both the cell nucleus and cytoplasm. Bioinformatics analysis indicates that the TaGRAS34-5A promoter contains a large number of hormone response elements and light response elements, and it is most closely related to TaSCL14, OsGRAS23, and AtSCL14 in terms of phylogenetic relationships, suggesting its potential function in responding to oxidative stress. Its expression is upregulated under high-temperature, ethylene (ETH), abscisic acid (ABA), and salicylic acid (SA) treatments, peaking at 4, 6, 0.5, and 12 hours post-treatment, respectively, with the most significant induction observed under heat stress and SA. Functional assays in yeast demonstrated that heterologous expression of TaGRAS34-5A enhances the heat tolerance of the yeast. The results of BSMV:VIGS transient silencing experiment showed that after the 42 ℃ high-temperature treatment, TaGRAS34-5A silenced plants exhibited decreased chlorophyll content, reduced POD enzyme activity, increased cellular peroxidation, and decreased heat tolerance compared to the control. Preliminary studies on the heat tolerance mechanism suggest that TaGRAS34-5A exhibits strong transcriptional self-activation activity.it may modulate wheat heat tolerance by interacting with proteins such as the bZIP family transcription factor HBP-1b and the E3 ubiquitin ligase hel2, thereby regulating cellular redox homeostasis and detoxification processes, positively influencing the heat tolerance of wheat.【Conclusion】TaGRAS34-5A is induced by heat, ABA, ETH, and SA, and its encoded protein is located in the nucleus and cytoplasm. It exhibits transcriptional activation activity. Heterologous overexpression of TaGRAS34-5A enhances the heat tolerance of Saccharomyces cerevisiae. Silencing TaGRAS34-5A in wheat plants increases cellular peroxidation, decreases chlorophyll content, and reduces heat tolerance. TaGRAS34-5A may regulate the heat tolerance of wheat by modulating cellular redox state and detoxification processes.

QTL Mapping of Quality Traits for A Peanut Germplasm SW9721-3 with Ultra-High Oil Content

YANG YongQing, HU PengJu, SONG YaHui, JIN XinXin, SU Qiao, WANG Jin

Scientia Agricultura Sinica. 2025, 58(4): 635-646. doi:10.3864/j.issn.0578-1752.2025.04.002

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【Objective】High quality is a critical objective in peanut breeding. However, the poor genetic base of cultivated peanuts has significantly limited the breeding efficiency for high-quality peanut. Therefore, unearthing valuable allelic gene resources associated with quality traits would provide basis for expanding genetic diversity of cultivated peanut germplasm resources.【Method】A wild peanut (Arachis villosa), PI 210553, carried diploid A type genome, was used as a donor parent for crossing with cultivated peanuts. Ultra-high oil peanut germplasm with wild ancestry was selected from the progeny, and a recombinant inbred line (RIL) population, derived from this germplasm and the cultivated variety Jihua 5, was used for genetic dissection and QTL mapping of four quality traits across three environments.【Result】Four ultra-high oil accessions of the SW9721 series were selected from the cross population between Yueyou 551 and PI 210553. Notably, SW9721-3 showcased an oil content of 62.50%, exceeding the national high-oil-content peanut standard by 7.5 percent points. Phenotypic analysis of the RIL population revealed that environmental influences caused variations in the mean values of quality traits from 11.02% to 40.80%. Correlation coefficients among these traits varied from 0.23 to 0.97 (P<0.001), with significant associations observed between oil and protein contents, and between oleic and linoleic acid contents. Furthermore, the phenotypic variation of quality traits was found to be between 3.78% and 10.61%, with heritability values above 0.6, absolute values of skewness and kurtosis were less than 1. These phenotypic values were approximately normally distributed, indicating that these traits are quantitatively inherited. In addition, a total of 12 QTLs were identified across the three environments, with LOD scores ranging from 3.26 to 17.82, accounting for 1.97% to 24.56% of the phenotypic variation. Particularly, The QTL locus qPOC_7 was consistently detected in all environments, with LOD scores between 6.01 and 17.82, explaining 4.59% to 24.56% of the phenotypic variation in both protein and oil content. The oil content increase was attributed to the allele from SW9721-3, highlighting the significant breeding value of this QTL. The physical position of flanking molecular markers for qPOC_7 suggested that the corresponding genes are located within a 180 kb interval from 426 363 to 606 659 bp at the end of chromosome 7. Within this candidate region, 22 annotated genes were identified, including two glucose metabolism- related genes, Ah.CKCA5J and Ah.805HV8, which are considered the high-potential candidate genes. 【Conclusion】In summary, this study successfully developed an ultra-high oil peanut germplasm, SW9721-3, and identified a major QTL locus, qPOC_7, with significant implications for peanut breeding.

Effects of Fallow Weeds Returning to the Field on Photosynthetic Matter Production and Yield of Rice

CHEN Ge, GU Yu, WEN Jiong, FU YueFeng, HE Xi, LI Wei, ZHOU JunYu, LIU QiongFeng, WU HaiYong

Scientia Agricultura Sinica. 2025, 58(4): 647-659. doi:10.3864/j.issn.0578-1752.2025.04.003

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【Objective】This study aimed to investigate the effects of different organic materials returning to the field on photosynthetic matter production and yield formation in an indica three-line hybrid rice population, to provide a theoretical basis for the rational utilization of straw resources and the improvement of rice yield. 【Method】 A field experiment was conducted in Matang Town, Yueyang City, Hunan Province, China from 2022 to 2023. Chuanzhongyou 464, a widely cultivated hybrid rice variety, was used as the test material. Six treatments were designed, including conventional nitrogen fertilizer (N), rice straw returning+conventional nitrogen fertilizer (RN), rape straw returning+conventional nitrogen fertilizer (ON), Chinese milk vetch returning+conventional nitrogen fertilizer (MN), fallow weed returning+conventional nitrogen fertilizer (WN), and no nitrogen fertilizer and no straw returning as control (CK). At various growth stages, the tiller dynamics, leaf area index, aboveground dry matter accumulation, SPAD value of flag leaves at heading stage, net photosynthetic rate, and yield and yield components of rice were measured under different modes of straw returning to the field. 【Result】Compared with N treatment, the effective panicles per hm², total spikelets per hm², and grain yield under WN treatment increased significantly by 13.65%, 16.85%, and 9.65%, respectively. There was a slight increase in spikelets per panicle and grain weight, while the seed setting rate decreased, but the difference was not significant. In terms of photosynthetic matter production characteristics, under WN treatment, the percentage of productive tiller increased significantly by 13.08%, and the SPAD value in the flag leaf, net photosynthetic rate, and stomatal conductance of flag leaves at heading stage were enhanced significantly by 3.57%, 9.16%, and 16.68%, respectively. Meanwhile, the dry matter accumulation in the aboveground part measured at mature stage increased by 13.79%, photosynthetic potential, crop growth rate, and dry matter accumulation in the aboveground part from heading to mature stage increased by 9.20%, 33.12%, and 33.21%, respectively, and the harvest index improved by 12.31%. In addition, the leaf area index, net assimilation rate, post-heading leaf and stem sheath output and export rate also showed an increase trend. Correlation analysis results indicated that leaf SPAD value, net photosynthetic rate and stomatal conductance significantly and positively correlated with effective panicles per hm², spikelets per panicle, total spikelets per hm², and grain yield. 【Conclusion】Fallow weeds returning to the field improved the photosynthetic characteristics of the rice at heading stage, enhanced the production capacity of photosynthetic matter after heading, promoted dry matter accumulation after heading stage, and optimized population quality. Simultaneously, by increasing the ratio of productive tiller, the suitable effective panicles per hm² were constructed and the total spikelets per·hm² increased, it thereby achieved higher grain yield.

Antioxidant Characterization of Nitrogen Application for Mitigating Potato Senescence Post-Flowering Under High Temperature Stress

SU Ming, LI FanGuo, HONG ZiQiang, ZHOU Tian, LIU QiangJuan, BAN WenHui, WU HongLiang, KANG JianHong

Scientia Agricultura Sinica. 2025, 58(4): 660-675. doi:10.3864/j.issn.0578-1752.2025.04.004

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【Objective】In order to explore the mechanism of nitrogen application under high temperature on the antioxidant characteristics of dryland potato in the mountainous area of southern Ningxia, and to elucidate the mechanism of nitrogen regulation, so as to provide the reference for the local development of nitrogen application measures favorable to alleviate high temperature stress. 【Method】 A 2-year field in situ experiment was conducted in Dazui Village, Haiyuan County, Ningxia, from 2020 to 2021, using a split-zone experimental design with four N application levels as the main zones, namely 0 (N0), 75 kg·hm^-2 (N1), 150 kg·hm^-2 (N2), and 225 kg·hm^-2 (N3), and two temperature gradients as the sub-zones, namely (35±2) ℃ (HT) and (30±2) ℃ (CK). The effects of post-flowering high temperature stress on potato leaf area index (LAI), relative chlorophyll content (SPAD), antioxidant properties, membrane lipid peroxidation products and non-enzymatic protective substances were analyzed. 【Result】 The 2-year results showed that potato LAI, SPAD, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) showed a decreasing trend from 30d-35d after flowering under the nitrogen fertilizer×temperature intercropping condition; at the same time, the cell membrane permeability, malondialdehyde content (MDA) and proline content (Pro) of potato leaves showed an increasing trend. Taking 35d after flowering as an example, after high temperature stress, potato LAI decreased by 11.22%-21.20%, SPAD decreased by 23.29%-26.05%, SOD decreased by 12.27%-16.87%, POD decreased by 13.69%-17.71% and CAT decreased by 13.80%-18.39% compared with room temperature; while cell membrane permeability, MDA content and Pro content increased significantly. Meanwhile, after high temperature stress, LAI and SPAD were significantly increased, while SOD, POD, CAT as well as yield reached the highest under N2 treatment (150 kg·hm^-2) compared with other treatments and N2 could also reduce cell membrane permeability and MDA content to alleviate high temperature hazards and increase potato yield. To further explore the correlation between potato yield and N application, a quadratic relationship between yield and N application was found, which led to the derivation of the corresponding parameter values for the economically optimal N application rates of 132-142 kg·hm^-2 (HT) and 185-210 kg·hm^-2 (CK). Pearson's correlation analysis showed that under high temperature stress, yield was only related to leaf LAI, POD, and CAT. LAI, POD and CAT reached significant positive correlation and significant negative correlation with MDA and Pro, while it did not reach significant level with SOD, SPAD and cell membrane permeability. Meanwhile, through the principal component analysis, it was found that after 2 years of high temperature stress, the composite scores of different nitrogen application levels were N2>N3>N1>N0. 【Conclusion】 The application of nitrogen at 150 kg·hm^-2 could continue to improve the physiological and antioxidant characteristics of potato leaves and to optimize its yield effectively, and it was also consistent with the theoretical estimation of 2 years of post-flowering high temperature. It was found that the N application rate of 150 kg·hm^-2 could continuously improve the physiological characteristics of potato leaves and effectively optimize the yield, and the difference was very small with the theoretically estimated 2-year economic optimum N application rate (132-142 kg·hm^-2). Therefore, the present experiment could also take 150 kg·hm^-2 as the recommended N application rate for safe potato production in Ningnan mountainous area to cope with the increasingly serious local high temperature hazard.

The Physiological Response of Longzhong Alfalfa to Exogenous Spermine Under Drought Stress

WANG WenJuan, SHI ShangLi, KANG WenJuan, DU YuanYuan, YIN Chen

Scientia Agricultura Sinica. 2025, 58(4): 676-691. doi:10.3864/j.issn.0578-1752.2025.04.005

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【Objective】Spermine (Spm) can improve the tolerance of plants to abiotic stresses and reduce the damage caused by abiotic stresses to plants. To investigate the effect of exogenous Spm on drought resistance of alfalfa under drought stress, to provide a theoretical basis for exogenous Spm to improve drought resistance of alfalfa, and then to provide a scientific basis for the application of utilizing exogenous Spm to enhance drought resistance of plants. 【Method】Longzhong alfalfa (Medicago sativa L. cv. Longzhong) was used as the experimental material, and polyethylene glycol 6000 was used to simulate drought stress. Under normal water and drought stress after the root application of 0.1, 0.5 and 1.0 mmol·L^-1 concentration of Spm, the change characteristics in Longzhong alfalfa were studied, including plant height, leaf relative water content (RWC), photosynthetic pigments content, gas exchange parameters, antioxidant enzymes activity, antioxidants content, superoxide anion ($O^{\bar{.}}_{2}$) content, hydrogen peroxide (H₂O₂) content, malondialdehyde (MDA) content and osmoregulatory substances content. 【Result】Under drought stress, exogenous Spm increased plant height and leaf RWC of Longzhong alfalfa, slowed down the degradation of chlorophyll a, chlorophyll b (Chlb), and carotenoids, elevated the net photosynthetic rate, transpiration rate, and stomatal conductance of leaves, reduced the concentration of intercellular carbon dioxide, enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), raised the ascorbic acid (ASA) content, ASA/ dehydroascobic acid (DHA) ratio, glutathione (GSH) content and GSH/ glutathione disulfide (GSSG) ratio, decreased DHA and GSSG content, enhanced ascorbate peroxidase and glutathione reductase activities, decreased $O^{\bar{.}}_{2}$ and H₂O₂ production, inhibited MDA accumulation, and increased proline and soluble sugar content. The results of principal component analysis and calculation of average affiliation function values of all physiological indexes under each treatment showed that Spm application under normal water conditions had no effect on the growth of alfalfa, whereas Spm application under drought condition improved the drought tolerance of Longzhong alfalfa under drought stress, with the best enhancement effect of 0.5 mmol·L^-1 Spm. A further redundancy analysis showed that 0.5 mmol·L^-1 Spm enhanced the drought tolerance of Longzhong alfalfa mainly by slowing down the degradation of leaf Chlb, increasing the ASA-GSH cycle and CAT activity.【Conclusion】Exogenous Spm could reduce the degradation of photosynthetic pigments, alleviate the non-stomatal restriction of drought stress on alfalfa leaves, increase the activities of SOD, POD and CAT, promote the ASA-GSH cycle, increase the content of osmoregulatory substances, reduce the production of $O^{\bar{.}}_{2}$ and H₂O₂, and decrease the degree of membrane lipid peroxidation, thus increasing the plant height and leaf RWC of Longzhong alfalfa under drought stress, thereby enhancing its drought resistance. Exogenous application of 0.5 mmol·L^-1 Spm was an effective method to improve the drought resistance of alfalfa under drought stress, and it could be applied to improve the drought resistance of alfalfa in dry areas for practical production.

Effects of Tomato-Rice Rotation on Physicochemical Properties and Microbial Communities of Soil with Continuous Cropping Obstacles in Cangnan, Zhejiang

WANG ShaoHua, SHEN NianQiao, CHU TianRan, WU YongHan, LI KangNing, SHI YanXia, XIE XueWen, LI Lei, FAN TengFei, LI BaoJu, CHAI ALi

Scientia Agricultura Sinica. 2025, 58(4): 692-703. doi:10.3864/j.issn.0578-1752.2025.04.006

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【Objective】Long-term continuous cropping of tomato in Cangnan, Zhejiang Province leads to a series of soil continuous cropping obstacles, such as soil acidification, secondary salinization, soil compaction and serious soil-borne diseases. In this paper, the effects of tomato-rice rotation on soil physicochemical properties, enzyme activity, microbial biomass and microbial community structure under long-term continuous cropping were analyzed, aiming to provide a method to alleviate soil continuous cropping obstacles and improve soil environment.【Method】Three treatments were set up and soil samples were collected. T1 was tomato continuous monoculture for 18 years, T2 was tomato continuous monoculture for 17 years followed by one year of tomato-rice rotation, and T3 was tomato continuous monoculture for 15 years followed by three years of tomato-rice rotation. Soil pH and EC values were measured by pH meter and conductivity meter. Soil physicochemical properties such as total carbon, total nitrogen and ammonium nitrogen were determined by high temperature combustion method, Kjeldahl nitrogen fixation method and potassium chloride solution leaching method. Soil enzyme activities were determined using enzyme activity kits. Total and harmful microorganism concentrations were detected using qPCR, and soil culturable microorganisms were counted using selective medium plate smear counting method. The MiSeq PE3000 high-throughput sequencing platform was used for sequencing, splicing and assembly, sequence comparison and functional annotation of the soil microgenome.【Result】There were significant differences in soil physicochemical properties among different treatments. One and three years of tomato-rice rotation treatments increased soil pH from 5.20 to 6.04 and 6.73, and reduced EC from 558 μS·cm^-1 to 417 and 445 μS·cm^-1, and increased C﹕N ratios from 9.16 to 10.45 and 10.74. Soil enzyme activities were increased in the rotations, with catalase, urease, polyphenol oxidase activities increased from 11.72 μmol·d^-1·g^-1, 10.76 μg·d^-1·g^-1, and 22.67 mg·d^-1·g^-1to 58.58 μmol·d^-1·g^-1, 142.48 μg·d^-1·g^-1, and 37.10 mg·d^-1·g^-1. After crop rotation, the microbial population and community structure changed, harmful microorganisms decreased. The content of soil culturable actinomycetes increased, the fungi/bacteria ratio decreased. Chloroflexi, Acidobacteria, and Proteobacteria increased significantly, and Actinobacteria decreased significantly. The contents of Ralstonia solanacearum, Pectobacterium carotovorum subsp. carotovorum, P. c. subsp. brasiliensis and Fusarium sp. decreased to 1.76×10³, 7.28×10², 3.94×10³ and 3.07×10³ copies/g in the three years of crop rotation. The potential functions of soil microorganisms changed after crop rotation, with an increase in the abundance of genes related to carbohydrate metabolism, energy metabolism, and other related pathways. Specifically, genes associated with carbohydrate metabolism in metabolism were up-regulated.【Conclusion】Tomato-rice rotation can improve soil acidification, salinization and soil element imbalance caused by long-term monoculture, increase soil C﹕N ratio and enzyme activity, reduce the occurrence of soil-borne diseases, transform the soil from fungal type to bacterial type, change the structure of soil microbial community, and promote soil microbial carbohydrate metabolism, which is important for improving soil continuous cropping obstacles.

Genome-Wide Identification and Expression Analysis of HMG Family Genes in Botrytis cinerea

ZHANG TianYu, LI Bai, ZANG JinPing, CAO HongZhe, DONG JinGao, XING JiHong, ZHANG Kang

Scientia Agricultura Sinica. 2025, 58(4): 704-718. doi:10.3864/j.issn.0578-1752.2025.04.007

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【Objective】High mobility group protein (HMG), as a widespread eukaryotic protein, plays an important role in the regulation of fungal growth and development. This study aims to identify the HMG family members of Botrytis cinerea through the genome-wide identification, and analyze their expression rules during the growth and development, infection and response to stress, so as to lay a foundation for further investigation of the function of this family gene in the growth and development and pathogenic mechanism of B. cinerea.【Method】HMMER software was used to screen out possible HMG gene IDs in B. cinerea protein database, and the online tools Pfam and SMART were used to verify the possible HMG family members. Using ProtParam, TBtools, MEME, MEGA, MCScanX and other tools, the physicochemical characteristics, chromosome localization, gene structure, phylogenetic tree and collinearity of B. cinerea HMG gene family members were analyzed. Based on transcriptome sequencing data of B. cinerea during conidial development and infection, the expression of HMG family genes in B. cinerea during conidial development and infection was analyzed by using TBtools software. In addition, the expression of HMG family genes in B. cinerea under NaCl, KCl, H₂O₂ and other stresses was observed by real-time quantitative PCR (qPCR).【Result】Nine HMG family members were identified in B. cinerea, the number of encoded amino acids ranged from 101 to 1076, and the molecular weight ranged from 11.48 to 120.64 kDa. They could be divided into HMGA and HMGB subfamilies, which had conserved HMG domain, distributed on six chromosomes, and had gene collinearity with Fusarium graminearum and Fusarium oxysporum. The expression pattern analysis showed that the expression levels of this family gene showed obvious changes during conidial development and B. cinerea infection. Among them, Bcin01g08840, Bcin07g04610, Bcin11g04340 and Bcin11g03320 continued to be highly expressed during conidial development. The expression levels of Bcin07g04610, Bcin11g04340, Bcin11g05790 and Bcin01g08840 were significantly down-regulated during B. cinerea infection. qPCR analysis showed that the gene expression of this family was significantly down-regulated after NaCl and KCl stress treatment.【Conclusion】B. cinerea contains nine HMG family members, which can be divided into HMGA and HMGB subfamilies, the members distribute on six chromosomes, and have different expression patterns in the process of conidial development, infection and stress response of the pathogen. It is speculated that the family genes play an important role in the growth and development, pathogenicity and stress response of B. cinerea.

The Variation Characteristics of Soil Organic Carbon Fractions Under the Combined Application of Organic and Inorganic Fertilizers

SHI Fan, LI WenGuang, YI ShuSheng, YANG Na, CHEN YuMeng, ZHENG Wei, ZHANG XueChen, LI ZiYan, ZHAI BingNian

Scientia Agricultura Sinica. 2025, 58(4): 719-732. doi:10.3864/j.issn.0578-1752.2025.04.008

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【Objective】This experiment was conducted to study the content characteristics of soil organic carbon components under long-term organic and inorganic fertilizers and the contribution of each component to the yield of winter wheat in dryland, with a view to providing an important theoretical basis and practical value for the increase of wheat yield and soil fertilization in Northwest dryland. 【Method】This study was based on a nine-year long-term positioning experiment, using a fissure design, with two organic fertilizer levels (M0 and M1) for the main treatment, and five nitrogen levels (N0, N75, N150, N225, N300) for the side treatment. The variation characteristics of winter wheat grain yield and soil organic carbon and its components were analyzed, including dissolved organic carbon (DOC), microbial biomass carbon (MBC), particulate organic carbon (POC), and mineral organic carbon (MOC), as well as the differences in the mass fractions of the components, and the contribution of each component of organic carbon to wheat grain yield was quantified. 【Result】With the increase of nitrogen application rate, the wheat grain yield increased first and then decreased, while the grain yield of wheat increased by 4.80% under organic fertilizers than that under chemical fertilizers alone. The highest yield (8 143.2 kg·hm^-2) was obtained under M1N150 treatment, which increased by 85.36% compared with M0N0 treatment, and this fertilizer application decreased by 75 kg·hm^-2 compared with the local conventional nitrogen application. After 9 years of continuous fertilizer application, the SOC content under M1N150 treatment was significantly increased by 103.30% compared than the soil at the initial stage of the experiment (2014). That is, the nitrogen rate of 150 kg·hm^-2 combined with 30 t·hm^-2 organic fertilizer would not only significantly improve the soil fertilizer cultivation effect, but also increase the winter wheat yield on the basis of reducing nitrogen application. The study of soil organic carbon fractions under different fertilization treatments showed that, compared with chemical fertilizers alone, the organic and inorganic fertilizers combined treatments increased the content of POC, DOC and MBC fractions as well as the proportion of POC fractions in SOC. The sensitivity indices of SOC and the contents of each organic carbon component showed that the soil active organic carbon components (DOC, MBC and POC) responded significantly to the organic and inorganic fertilizer treatments, among which the POC and DOC components were the most sensitive to the response of farmland management measures. The correlation analysis between soil organic carbon components and wheat yield showed that POC, DOC and MBC components had positive effects on yield increase; the results of Random Forest Analysis (RFA) further proved that the POC and DOC components contributed more to wheat yield under the combined application of organic and inorganic fertilizers. Therefore, it could be inferred that the increase of crop productivity and soil fertility by organic and inorganic fertilization was mainly achieved by increasing the DOC and POC content of organic carbon in the soil. 【Conclusion】When 150 kg·hm^-2 of nitrogen was applied with 30 t·hm^-2 of organic fertilizer, it was more conducive to the enhancement of the content of soil organic carbon and reactive organic carbon fractions in wheat fields in the drylands of Northwest China, which in turn improved the yield of winter wheat in dryland, with the DOC and POC fractions contributing the most to the yield of wheat.

Evaluation of Carbon Footprint and Economic Benefit of Different Tobacco Rotation Patterns

ZHENG Yu, CHEN Yi, TI JinSong, SHI LongFei, XU XiaoBo, LI YuLin, GUO Rui

Scientia Agricultura Sinica. 2025, 58(4): 733-747. doi:10.3864/j.issn.0578-1752.2025.04.009

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【Objective】This study aimed to clarify the economic benefits and carbon emission characteristics under different cropping patterns, so as to provide a scientific basis for the selection of regional dominant cropping rotation patterns.【Method】Based on the survey data of farmers in Xiangcheng County, Xuchang City, central Henan Province from 2018 to 2022, this study calculated the carbon emissions of four different rotation patterns: cereal-vegetable-tobacco (wheat-pepper-wheat-pepper-wheat-pepper-tobacco), wheat-maize-tobacco (wheat-maize-wheat-maize-wheat-maize-tobacco), cereal-soybean-tobacco (wheat-maize-wheat-soybean- wheat-soybean-tobacco), and cereal-sweet potato-tobacco (wheat-maize-wheat-sweet potato-wheat-sweet potato-tobacco). The life cycle assessment was used to calculate the carbon emissions and carbon footprint of these four cropping models. This study also clarified their composition and comprehensively evaluated the economic and ecological benefits of four crop rotation patterns by considering input costs and harvest output values.【Result】(1) The output value and profit ranking under different planting patterns were as follows: cereal-vegetable-tobacco>cereal-sweet potato-tobacco>wheat-maize-tobacco>cereal-soybean-tobacco. Additionally, the total cost of the grain-vegetable- tobacco model was significantly higher than that of the other three patterns. (2) The carbon footprint per unit area under different rotation patterns was as follows: wheat-maize-tobacco>cereal-sweet potato- tobacco>cereal-soybean-tobacco>cereal-vegetable- tobacco, with the value of 32 391.10, 31 042.64, 30 583.80, and 26 524.57 kg·hm^-2, respectively. The carbon footprint per unit yield for different crop rotation patterns followed this order: cereal-soybean- tobacco (0.51 kg·kg^-1), wheat-maize-tobacco (0.51 kg·kg^-1), cereal-sweet potato-tobacco (0.39 kg·kg^-1), and cereal-vegetable-tobacco (0.29 kg·kg^-1). The carbon footprint per unit output value of cereal-soybean-tobacco, wheat-maize-tobacco, cereal-sweet potato-tobacco, and cereal-vegetable-tobacco systems were 0.17, 0.17, 0.13, and 0.10 kg/yuan, respectively. (3) Fertilizer accounted for 50.6%, 56.4%, 57.2%, and 57.0% of carbon emissions in cereal-vegetable-tobacco, wheat-maize-tobacco, cereal-soybean-tobacco, and cereal-sweet potato-tobacco, respectively, making it the primary contributor to carbon emissions in each rotation pattern. The second largest contributor was tobacco curing electricity, which accounted for 15.2%, 14.5%, 13.5% and 13.0% in the above rotation patterns. 【Conclusion】Under the four crop rotation patterns, the cereal-vegetable-tobacco model demonstrated high economic benefits and low carbon emission. However, the input cost of this model was the highest, while the output value of tobacco was the lowest. Therefore, reducing labor input with improving tobacco benefits was crucial for promoting this rotation pattern in major tobacco producing areas. Tobacco of cereal-sweet potato-tobacco model had the highest output value and the lowest cost, but the high carbon footprint of this rotation pattern might be a constraint to the promotion. Fertilizer and tobacco curing electricity were the main sources of carbon emissions for each rotation patterns. Therefore, improving fertilizer utilization rate, and promoting clean energy barns to reduce the carbon footprint of tobacco leaf curing were the keys to reduce carbon emissions from various crop rotation patterns.

Suitability Evaluation of Late Maturing Citrus in Dongpo District, Meishan City, Sichuan Province Based on Maximum Entropy Model (MaxEnt) and Geographic Information System (GIS)

YAO Li, DENG ChunXiu, TANG Biao, WANG Hong, WU YueYing, ZHANG Qi, LI YuanHong, LIU ZhongYou, LU ChangAi, LIN ChaoWen

Scientia Agricultura Sinica. 2025, 58(4): 748-758. doi:10.3864/j.issn.0578-1752.2025.04.010

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【Objective】Carrying out the suitability evaluation of late-maturing citrus in Dongpo District, Meishan City, can provide scientific references for the reasonable cultivation and optimization of production space of local late-maturing citrus. 【Method】Based on 14 climatic indicators, 3 topographic indicators and 22 soil physicochemical indicators, as well as data from 372 sampling points of late-maturing citrus planting areas, the article utilizes the Maximum Entropy Model (MaxEnt) and Geographic Information System (GIS) in combination with correlation analysis to screen key environmental factors affecting the distribution of late-maturing citrus in Dongpo District, Meishan City, Sichuan Province, and constructs a model for evaluating the suitability of late-maturing citrus planting. The model was constructed to evaluate the suitability of late-maturing citrus planting.【Result】The MaxEnt model was very effective, and the area under the curve (AUC) in the characteristic working curve (ROC curve) of the subjects reached 0.843, which was highly accurate and reliable. The MaxEnt analysis showed that the average annual air temperature, elevation, total phosphorus, pH, average annual relative humidity, pulverization, annual precipitation, annual sunshine hours, autumn precipitation, soil exchangeable calcium, total potassium and organic matter were the main environmental factors affecting the distribution of late-maturing citrus; the cumulative contribution of climatic variables affecting the distribution of citrus was 43.0%, of which mean annual temperature (22.9%), mean annual relative humidity (7.0%) and annual precipitation (4.6%) were the three most important climatic factors affecting the distribution of late-maturing citrus; the cumulative contribution of soil variables affecting the distribution of late-maturing citrus was 41.3%, of which total phosphorus (13.8%), pH (10.4%) and chalkiness (5.7%) were the three most important soil factors affecting the distribution of late-maturing citrus; elevation was a topographic factor affecting the distribution of citrus, with a contribution rate of 15.7%. By statistically analyzing the parameter values of each dominant environmental factor in different levels of distribution areas, combining the response curves of each ecological factor, the field survey and the review of the literature, a comprehensive analysis was made to conclude that the basic climatic and topographical environmental requirements for the distribution of late-maturing citrus are as follows: the average annual air temperature is about 17 ℃, the average annual relative humidity is 80%, the annual rainfall is about 1 300 mm, the annual sunshine hours are 1 350 h, and the rainfall in autumn is about 390 mm, altitude of about 400-700 m; soil pH of 4.5-7.5, while the soil nutrient status is affected by the combination of standing conditions and cultivation and management measures. Of the 34 673.29 hm² of gardenable land in the region, 23.51%, 61.37% and 15.12% were in the dominant, suitable and unsuitable zones, respectively.【Conclusion】This study objectively reflects the changing characteristics of late-maturing citrus planting suitability, suitable area and distribution in Dongpo District under the influence of climate, topography and soil factors.

Dynamics of Fruit Hollowness and Browning and Associated Lignin Accumulation and Its Genome-Wide Identification of Ps4CL Gene Family in Huangguan Plum

LI JianKun, PENG Chao, ZHANG ZiYang, LIANG Xi, WEI MingKang, YANG Qiang, LI BinQi, Muhammad Moaaz Ali, Viola Kayima, CHEN FaXing, DENG HongHong

Scientia Agricultura Sinica. 2025, 58(4): 759-778. doi:10.3864/j.issn.0578-1752.2025.04.011

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【Objective】The objective of this study was to examine the temporal dynamics of hollowness and browning (HB) in Nai plum during its fruit development and ripening, to evaluate the relationship between HB and lignin accumulation, and to identify the 4CL gene family in Huangguan plum, a critical gene in the lignin biosynthetic pathway, at the whole-genome level. 【Method】Huangguan plum fruits were sampled at 10, 25, 40, 55, 70, and 80 days after full bloom to investigate the dynamics of HB. Anatomical changes in fruit structure induced by HB were observed by using paraffin sectioning. Lignin monomers were identified by using gas chromatography-mass spectrometry (GC-MS), while lignin content and associated enzyme activities were measured via UV-spectrophotometer. Gene expression was quantitatively analyzed through quantitative reverse transcription polymerase chain reaction (qRT-PCR). The genome-wide identification of the 4CL gene family was conducted utilizing tools, such as NCBI, TBtools, MEME, MEGA11, SOPMA, SWISS-MODEL, and PlantCARE. 【Result】HB initiated at the veraison stage, with the HB rate reaching 37.73% at the mature stage. Additionally, the fruit shape index of HB plum was significantly higher than that of non-HB plum. Histological analysis using paraffin section showed that the HB pulp was stained red by Saffron-O and Fast Green Stain and the HB area increased as the fruit developed. The total lignin content exhibited a gradual increase during fruit development and ripening, with the total lignin content in HB fruit being significantly higher than that in non-HB fruit (P≤0.001). The lignin monomers in HB fruit were predominantly composed of guaiacyl lignin (22.6%) and syringly lignin (10.8%). Both 4CL and PAL enzyme activities were significantly elevated in HB fruit compared with non-HB fruit during fruit development and ripening period. Expression levels of eight Ps4CLs were higher in HB fruit compared to non-HB fruit, with similar trends observed for two PsPALs which showed slightly higher expression in HB fruit during veraison stage. Ps4CL1, Ps4CL2, Ps4CL7, and Ps4CL8 were potentially implicated in the biosynthesis of lignin of plum fruit. The Ps4CL gene family characterized by a significant number of cis-acting elements associated with plant endogenous hormones. Collinearity analysis indicated the presence of two and eight pairs of 4CL homologous genes between plum and the Arabidopsis and peach, respectively. Weighted gene co-expression network analysis (WGCNA) identified core hub genes (Ps4CL5, Ps4CL7, and Ps4CL8), each exhibiting co-expression with at least 30 other genes. The qRT-PCR results demonstrated that Ps4CL genes exhibit tissue-specific and temporally specific expression profiles. 【Conclusion】This paper identified the critical period for HB occurrence (the veraison stage) and characterized the dynamics of lignin content, monomer type, enzyme activity, and related gene expression in Huangguan plum. WGNCA highlighted key genes involved in the lignin biosynthetic pathway in Huangguan plum, which laid a foundation for further exploration of regulatory mechanisms, control strategies, and related molecular mechanisms of HB in plum fruit.

Regression Analysis of the Differences in the Bioaccessibility of Six Nutrients in the Flesh of Melons from Different Production Areas and Their Relationship with Soil Element Content

WANG XiaoTing, SHEN Qi, PAN JuXiu, MA Lei, HE WeiZhong, WANG Cheng

Scientia Agricultura Sinica. 2025, 58(4): 779-791. doi:10.3864/j.issn.0578-1752.2025.04.012

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【Objective】This study aimed to clarify the differences in the bioaccessibility of four sugars and two vitamins in melon pulp from different producing areas, and to explore the influence of soil element content on the formation of this difference, so as to provide suggestions and dietary references for melon cultivation and consumers.【Method】Taking the pulp of 30 samples of the same variety of melon Nasimi and the corresponding rhizosphere soil from Turpan, Hami, Yining producing areas as the research object, the content of reducing sugar, fructose, glucose, sucrose, VB₁, VB₂ and 22 elements (such as Fe, Mn, and Zn) in soil samples were analyzed with test equipment, such as high-performance liquid chromatograph (HPLC) and inductively co,upled plasma mass spectrometer (ICP-MS). In vitro simulated digestion was used to detect the content of related nutrients, and the bioaccessibility of nutrients was calculated. Statistical analysis methods such as correlation analysis, orthogonal partial least squares regression PLS-DA, and multiple linear regression analysis were used to compare the differences in the bioaccessibility of the above four sugars and two vitamins in melon pulp from three producing areas, and to explore the influence of soil element content on this difference.【Result】The bioaccessibility of reducing sugar and fructose in Turpan melon pulp was 118.35% and 113.02%, respectively, which was higher than that in the other two producing areas. The bioaccessibility of glucose in Hami melon pulp was higher than that in the other two producing areas, reaching 195.27%. The content and bioaccessibility of VB1 and VB2 in Yining melon pulp were higher than those in the other two producing areas. The results of correlation analysis showed that the bioaccessibility of reducing sugar in melon pulp was significantly negatively correlated with the contents of seven elements (Cr, Ni, Sn, Sb, Ba, Tl, and Pb) in soil, and positively correlated with the content of B element. The bioaccessibility of glucose was negatively correlated with the content of Be, Al, Sr, Sn, Sb, Ba, Tl, and Pb in soil, and positively correlated with the content of Ti, V, Co, Cu, As, Se, and Mo. The bioaccessibility of VB₁ was positively correlated with the content of Cr, Co, Ni, Cu, B, and Sr in soil. The bioaccessibility of VB₂ was positively correlated with the content of Al and Sr in soil. Regression analysis results showed that soil element content was an important factor affecting the bioaccessibility of four nutrients in melon pulp, and the influence degree was in the order of glucose > VB₁ > reducing sugar > VB₂. The content of Tl, V, Zn, Sn; Al, Zn, Sb, Tl, Fe, Mn, Zn; Al, V, Sr, Cd, and Ba in soil were important factors affecting the bioaccessibility of glucose, reducing sugar, VB₁, and VB₂ in melon pulp.【Conclusion】The technology for improving the bioaccessibility of glucose, reducing sugar, VB1, and VB2 in melon pulp could be explored by regulating the content of elements, such as Tl, V, Zn, Sn; Al, Zn, Sb, Tl, Fe, Mn, Zn; Al, V, Sr, Cd, and Ba in soil.

Research Progress on the Roles and Mechanisms of Autophagy Involved in Porcine Reproductive and Respiratory Syndrome Virus Infection

LI Rui, LIANG Yue, BAI Yang, ZHANG GuiYue, WANG NanNan, QIAO SongLin, ZHANG GaiPing

Scientia Agricultura Sinica. 2025, 58(4): 792-801. doi:10.3864/j.issn.0578-1752.2025.04.013

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Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped single-stranded positive sense RNA arterivirus. Infection by PRRSV results in porcine reproductive and respiratory syndrome (PRRS), characterized by abortions, stillbirths, weak-born piglets, and mummified fetuses in sows as well as respiratory diseases in pigs of all ages. PRRSV has caused tremendous economic losses to the global swine industry. Comprehensive investigation on PRRSV infection would be beneficial for the prevention and control of PRRS, and contribute to the high-quality development of the swine industry. Autophagy is a lysosome-dependent degradation and recycling process for abnormal proteins, damaged organelles, and invading pathogens. It comprises macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA),among which macroautophagy has been most extensively investigated so far, and is classified as non-selective autophagy and selective autophagy. Autophagy plays crucial roles in maintaining cellular homeostasis, controlling the quality of organelles, and transporting intracellular materials. Furthermore, autophagy is involved in diverse diseases, such as neurodegenerative diseases, autoimmune diseases, and cancers. Moreover, autophagy is a potent defense response to eliminate invading viruses, while viruses have evolved multiple strategies to subvert the autophagic degradation or even exploit autophagy for their own benefits. This review systematically scrutinized recent research progress on the roles and mechanisms of autophagy involved in PRRSV infection, and elaborated that autophagy played a dual role during PRRSV infection. On the one hand, PRRSV non-structural and structural proteins triggered macroautophagy (including reticulophagy, mitophagy, aggrephagy, and lipophagy) and CMA via a variety of mechanisms; in turn, macroautophagy and CMA facilitated PRRSV infection by participation in viral replication, antagonism of apoptosis, and inhibition of host immunity. On the other hand, selective autophagy (eg., reticulophagy) and CMA suppressed PRRSV infection by degradation of viral non-structural proteins and activation of antiviral immune responses. This review actually deepend the understanding of PRRSV infection from the autophagy perspective. In addition, this review summarized and raised certain controversial or unsolved issues, which provided clues for future studies on the roles and mechanisms of autophagy involved in PRRSV infection; in the meantime, this review illustrated and supported potential targets against autophagy for developing antiviral interventions to restrain PRRS epidemics.

Effect of VdF3′5′H2 Overexpression on the Accumulation of Anthocyanin Composition in Spine Grape Cells

GUO AoLin, LIN JunXuan, LAI GongTi, HE LiYuan, CHE JianMei, PAN Ruo, YANG FangXue, HUANG YuJi, CHEN GuiXin, LAI ChengChun

Scientia Agricultura Sinica. 2025, 58(4): 802-818. doi:10.3864/j.issn.0578-1752.2025.04.014

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【Objective】This study aims to clone the key gene VdF3′5′H2 involved in anthocyanin biosynthesis and analyze its regulatory role of anthocyanin biosynthesis and component content in spine grape (Vitis davidii Foëx) cells, thereby providing technical support and a theoretical foundation for constructing spine grape cells with high-yield anthocyanin production and enabling targeted regulation of anthocyanin biosynthesis.【Method】Using spine grape cells as materials, the VdF3′5′H2 was cloned and a plant expression vector was constructed. This vector was subsequently transformed into spine grape cell, with positive cell lines were screened in antibiotic media and identified through fluorescence observation and PCR analysis. The phenotypic characteristics of the transgenic spine grape cells were analyzed, along with quantification of anthocyanin, flavonoid, and proanthocyanidin. Additionally, the metabolite profile of anthocyanins was detected by UPLC-MS/MS. The expression levels of genes associated with anthocyanin biosynthesis were assessed using real-time quantitative PCR (RT-qPCR), and a comprehensive analysis of gene expression and differential metabolites was carried out by the O2PLS method.【Result】The open reading frame (ORF) of VdF3′5′H2 gene is 1 527 bp, encoding a protein comprising 508 amino acid residues. The VdF3′5′H2 protein exhibits high homology with its counterparts in related plants within the same family, containing the CYP75 conserved domain, a heme-binding site, and three characteristic conserved motifs. Phylogenetic analysis revealed that F3′H and F3′5′H cluster within the same branch, implying an evolutionary relationship from F3′H to F3′5′H, with VdF3′5′H2 positioned at a more ancestral state compared to VdF3′5′H1. Furthermore, subcellular localization indicated that the VdF3′5′H2 protein resides in the cytoplasm. The contents of anthocyanins, flavonoids and proanthocyanidins in two transgenic spine grape cells overexpression VdF3′5′H2 showed a significant increase compared to the wild type control. The content ratio of delphinidin-anthocyanins in the two transgenic cells increased to 7.82% (T6) and 14.32% (T10), respectively, Petunidin-anthocyanins increaseed to 7.30% and 10.16%, respectively, while the content ratio of malvidin- anthocyanins increased to 58.08% and 42.30%, respectively. In contrast, the content ratios of the three types of anthocyanins in the wild-type cells were 1.92%, 1.48%, and 8.49%, respectively. Additionally, the content ratios of cyanidin- and paeonidin-anthocyanins in the two transgenic cells decreased significantly. Overexpression of VdF3′5′H2 led to the downregulation of PAL, CHS, CHI and F3H expression levels, while upregulating the expression levels of VdF3′5′H1, LDOX and UFGT genes. The comprehensive analysis integrating gene expression data with differential metabolites showed that the overexpression of VdF3′5′H2 modulated the transcriptional regulation of genes involved in anthocyanin biosynthesis, thereby influencing both the synthesis pathways and accumulation patterns of various types of anthocyanins.【Conclusion】The overexpression of VdF3′5′H2 gene significantly enhanced the biosynthesis and accumulation of anthocyanins in spine grape cells, by regulating gene expression and altering the metabolic flux of the anthocyanin biosynthesis pathway, there by modulating the composition and content ratio of antocyanins in spine grape cells.

Effect of FGF5 and FGF21 on Proliferation of Dermal Papilla Cells in Cashmere Goat

WANG Niu, SHI XinRan, ZHANG WeiDong, WANG Xin

Scientia Agricultura Sinica. 2025, 58(4): 819-830. doi:10.3864/j.issn.0578-1752.2025.04.015

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【Background】The hair follicle cycle in goat is divided into three phases: the growth phase (anagen), the regression phase (catagen), and the resting phase (telogen). Fibroblast growth factor 5 (FGF5) and fibroblast growth factor 21 (FGF21) are important regulators in goat hair cycle. However, the precise localization of FGF5 and FGF21 in goat skin tissue and their regulatory mechanisms remain unclear during hair follicle cycle transition. Dermal papilla cells (DPCs) located in the dermal papilla region, are essential for hair follicle development. When DPCs lose their function, hair follicle remains in the telogen, leading to hair loss.【Objective】Therefore, this study aimed to identify the expression and localization of FGF5 and FGF21 in Cashmere goat skin tissue, understand their roles in DPCs, and to analyze their regulatory mechanisms. This would enrich the knowledge of fibroblast growth factor family proteins in hair follicle cycle regulation and provide a theoretical basis for further elucidating the molecular mechanisms of Cashmere goat hair follicle cycle transition. 【Method】Real-time quantitative PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression levels of FGF5, FGF21 and their receptor 1 (FGFR1) in the skin tissue of Cashmere goats at anagen and telogen. Single-cell transcriptome sequencing data were analyzed to determine the expression and localization of FGF5, FGF21 and FGFR1, which were validated through immunofluorescence methods. Adenovirus was explored to overexpress FGF5 and FGF21 in DPCs. The effects on DPCs proliferation were measured using MTT, EdU and flow cytometry. The expressions of FGFR1 and proliferation-related genes were detected using RT-qPCR, immunofluorescence and western blotting methods. RT-qPCR, immunofluorescence and western blotting were used to investigate the effect of FGF5 and FGF21 on the proliferation of DPCs. 【Result】FGF5, FGF21 and FGFR1 exhibited higher expression levels in anagen than that in telogen, all of which were located within DPCs. Both FGF5 and FGF21 restrained DPCs viability, which significantly reduced the number of EdU-positive cells. FGF5 decreased the percentage of DPCs in G2 and S phases, while FGF21 specifically affected the G2 phase. The expression of FGFR1 was significantly increased following the overexpression of FGF5 and FGF21. Moreover, the expressions of cell proliferation markers KI67 and PCNA were also significantly inhibited with the overexpression of FGF5 and FGF21. Additionally, the expression of β-catenin, a key component of Wnt/β-catenin signaling pathway, was reduced. Thereby the activity of transcription factors TCF3 and JUN was inhibited, which were the downstream effectors of Wnt/β-catenin signaling pathway. Thus the inhibition further led to the decreased expression of proliferation-related genes MYC and CYCLIND1. 【Conclusion】FGF5 and FGF21 played a critical role as regulatory factors in controlling the biological function of DPCs. This study provided the first evidences that FGF21, similar to FGF5, restrained the proliferation of DPCs by suppressing the Wnt/β-catenin signaling pathway. These findings contributes to our understanding of how fibroblast growth factors influence hair follicle cycle transition, particularly in the context of Cashmere goat hair growth and cashmere production.

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