Table of Content

01 March 2024, Volume 57 Issue 5

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Seedling Characterization and Genetic Analysis of Low Phosphorus Tolerance in Shanxi Varieties

WEI NaiCui, TAO JinBo, YUAN MingYang, ZHANG Yu, KAI MengXiang, QIAO Ling, WU BangBang, HAO YuQiong, ZHENG XingWei, WANG JuanLing, ZHAO JiaJia, ZHENG Jun

Scientia Agricultura Sinica. 2024, 57(5):  831-845.  doi:10.3864/j.issn.0578-1752.2024.05.001

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【Objective】In arid and semi-arid regions, the water and nutrients are scarce in the soil. The phosphorus use efficiency between different wheat genotypes varies greatly. Therefore, identification of low phosphorus-tolerant germplasm and mapping of related loci is helpful for genetic improvement of wheat. 【Method】Using 282 Shanxi wheat varieties as materials, twelve seedling morphological indicators were investigated under three phosphorus concentrations, including SDW, RDW, DW, SFW, RFW, FW, MRL, TRL, RS, RV, RD, and RN. Principal component analysis, membership function analysis, and cluster analysis were used to comprehensively evaluate the low phosphorus tolerance characteristics of different varieties at the seedling stage. On this basis, the trait evolution trend and biomass allocation at seedling stage were analyzed. At the same time, GWAS was used to identify significant loci related to the low phosphorus-related traits. 【Result】The response of different traits to low phosphorus at the seedling stage was different. Lower phosphorus concentrations led to changes in biomass allocation strategy, and shoot growth was less affected by change in phosphorus concentrations than root growth. The decrease in phosphorus concentration inhibited the growth of shoot, and SDW and SFW were significantly reduced. In contrast, low phosphorus promoted root growth, and the indicators of RDW, RFW, MRL, TRL, RV and RN increased significantly. According to the correlation analysis between D-value and morphological indicators, it was found that MRL and RD could be used as selection indicators for low phosphorus tolerance at seedling stage. Based on D-value clustering analysis, 9 low phosphorus tolerant varieties were selected, including Jinmai 46, Jinmai 61, Youmangdahongjing, Hongtumai, Hongheshang, Baikehong, Baixianmai, Huoshaotou, Baishanmai. Analysing trends in trait evolution showed that cultivars were not directly selected for their ability to tolerate low phosphorus. The ability to tolerate low phosphorus decreased first and then increased over time. Before 2010, there was a decreasing trend in the ability of varieties to tolerate low phosphorus, and after 2010, there was an increase in the ability of varieties to tolerate low phosphorus. GWAS stably detected eight loci with R²>10% in three environments, in which 1A_545074550, 2B_489279799, 6A_166899658 and 6A_273060644 were not reported previously.【Conclusion】The MRL and RD can be used as selection indicators for low phosphorus tolerance at seedling stage. A total of nine varieties were selected through comprehensive evaluation of ability in Shanxi wheat to tolerate low phosphorus during seedling stage. Association analysis detected four novel loci associated with low phosphorus tolerance on chromosomes 1A, 2B and 6A, and the results provide germplasm resources and QTL for future low phosphorus tolerance wheat breeding.

Current Situation of Breeding and Popularization of Short-Growth- Period Winter Rapeseed Varieties for Rice-Rice-Rapeseed Mode

LI RongDe, HE Ping, LUO LiXia, SHI MengYa, HOU Qian, MA ZhenGuo, GUO RuiXing, CHENG HongTao

Scientia Agricultura Sinica. 2024, 57(5):  846-854.  doi:10.3864/j.issn.0578-1752.2024.05.002

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The self-sufficiency rate of edible vegetable oil is less than 31 percent in China, with a high degree of import dependence. Rapeseed is the only winter oilseed crop with a wide range of suitable planting region, and it is an important source of edible vegetable oil in China. Planting more rapeseed is an important measure to guarantee national edible oil supply security. Making full use of the winter fields in the southern double cropping rice area to promote “rice-rice-rapeseed” production is an important approach to expand the planting area of rapeseed. The area suitable for the “rice-rice-rapeseed” production mode is mainly distributed in the double cropping rice area of Hunan, Jiangxi, Guangxi and Hubei provinces in China, with a potential area of about 1.87 million hm². According to the conditions of temperature and light resources, three suitable areas for the “rice-rice-rapeseed” production include the ample area, tightly balanced area and the constrained area. All the areas require early-maturing rapeseed varieties with a growth period of around 180 days, which are suitable for being sown in mid- to late October and harvested in mid- to late April. Among a total of 75 new rapeseed lines participated in the early-maturity group of the national rapeseed variety trials from 2013 to 2022, the average growth period ranged from 169.3 to 185.5 days, and the average yield was 1 635.90-2 228.55 kg·hm^-2, with 22 varieties out yielded the check variety. There are 72 early-maturing winter oilseed rape varieties with a growth period less than 190 days were registered by the end of May 2023. These varieties are suitable to be used in the “rice-rice-rapeseed” mode, and most of them are hybrid varieties with low erucic acid and low glucosinolate quality. 11 varieties, Yangguang 131, Fengyou 730, Fengyou 320, Fengyou 847, Ganyouza 906, Shengguang 127, Xiangyou 420, Jingyou 69, Fengyou 112, Huayouza 652, Ganyouza 1009, are the most promoted and applied early-maturing winter rapeseed varieties in the “rice-rice-rapeseed” production area, with more than 135 hm² each in 2022. The present varieties can basically meet the early-maturation demand in the ample area. In the tightly balanced and constraint areas, however, the growth period of these varieties is too long. To expand the production and efficiency of rapeseed industry, it is urgently needed to strengthen the policy and financial security in the future, carry out joint breeding projects for short-growth-period winter rapeseed varieties to further improve the yield in the ample area and shorten the growth period in the tightly balanced and constrained areas. Meanwhile, to strengthen the research and promotion of supporting cultivation technology for elite varieties, match well early rice, late rice, and rapeseed varieties are also good measures to support the expansion of rapeseed production in the “rice-rice-rapeseed” production area. In addition, improving agricultural socialized services, expanding agricultural insurance and increasing subsidies for rapeseed planting to ensure production benefit will enhance the farmers’ enthusiasm for the “rice-rice-rapeseed” production.

Identification of the Bru1 Genomic Region for Brown Rust Resistance and Functional Analysis of Candidate Resistance Genes in Sugarcane

WU QiBin, XIE WanJie, ZHONG Hui, FENG ChunYan, PAN HaoRan, QI YiYing, ZHANG JiSen, WANG HengBo

Scientia Agricultura Sinica. 2024, 57(5):  855-867.  doi:10.3864/j.issn.0578-1752.2024.05.003

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【Objective】Sugarcane brown rust, caused by Puccinia melanocephala H. Sydow & P. Sydow, is one of the most destructive fungal diseases in sugarcane industry, leading to a reduction in sucrose content by 10% to 36%. Previous studies revealed that the major gene(s) for brown rust resistance was located in the Bru1 genomic region. The cloning of crucial candidate genes and their functional investigation should provide important candidate gene resources for breeding new sugarcane cultivars resistant to brown rust.【Method】In this study, the contig-level genomic information of sugarcane cultivar ROC22 was obtained by utilizing the PacBio Sequel Ⅱ sequencing platform, and the Bru1 region associated with brown rust candidate resistance gene was identified, annotated, cloned, and analyzed for tissue specificity, expression patterns in resistant and susceptible sugarcane cultivars, subcellular localization, and transient overexpression.【Result】The results demonstrated that, firstly, using tightly associated molecular markers R12H16 and 9O20-F4 within the Bru1 region, a total of 33 genes were annotated from this region, and five candidate resistance genes (Brrg99, Brrg103, Brrg108, Brrg115, and Brrg116) were selected based on the typical/conserved domains of the resistance genes. Secondly, the full-length sequence cDNA sequence of the Brrg116 gene with an open reading frame of 729 bp and encoding 242 amino acid residues, was cloned from sugarcane cultivar ROC22. The gene sequence was aligned with the genomic databases of Saccharum spontaneum, S. officinarum, and the closely related diploid species sorghum. A high degree of sequence similarity was observed between S. spontaneum and S. officinarum, exceeding 98%. In contrast, its similarity with sorghum was 93.77%. Phylogenetic tree analysis suggests that this gene may originate from S. spontaneum species during sugarcane domestication. qRT-PCR analysis showed its constitutive expression in various tissues of sugarcane cultivars, particularly with the highest expression level in the +1 leaf, which was 5.2 times higher than in the bud. Furthermore, significantly differential expression of Brrg116 was observed at 6 h and 72 h post-inoculation with the brown rust pathogen in resistant and susceptible sugarcane cultivars. Subcellular localization analysis indicated that the encoded protein of this gene was located on the cell membrane. Finally, the Brrg116 gene was transiently overexpressed in Nicotiana benthamiana leaves, followed by inoculation with Fusarium solani var. coeruleum. The color of Nicotiana benthamiana leaves showed a gradual deepening phenotype by 3,3'-diaminobenzidine (DAB) staining. The genes related to hypersensitive response, salicylic acid, and ethylene synthesis had a sustained upregulation pattern, evidencing that the expression of these genes can enhance plant disease resistance.【Conclusion】Brrg116 was constitutively expressed in different sugarcane tissues and in response to brown rust pathogen infection, it showed a rapidly induced expression pattern in the resistant sugarcane cultivar. Overexpression of Brrg116 could trigger defense responses through hormone signaling pathways such as salicylic acid and ethylene. It is thus hypothesized that this gene may play a significant regulatory role in enhancing plant disease resistance.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Nitrogen Fertilizer Reduction and Postponing for Improving Plant Photosynthetic Physiological Characteristics to Increase Wheat- Maize and Annual Yield and Economic Return

ZHAO KaiNan, DING Hao, LIU AKang, JIANG ZongHao, CHEN GuangZhou, FENG Bo, WANG ZongShuai, LI HuaWei, SI JiSheng, ZHANG Bin, BI XiangJun, LI Yong, LI ShengDong, WANG FaHong

Scientia Agricultura Sinica. 2024, 57(5):  868-884.  doi:10.3864/j.issn.0578-1752.2024.05.004

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【Objective】Under the traditional nitrogen (N) fertilizer application technology in Huang-Huai-Hai Plain, insufficient soil N supply in the late growth stages of the wheat-maize double cropping system has resulted in significant yield reduction issues. In this study, the effects of N fertilizer reduction and postponing on plant photosynthetic physiological characteristics, aboveground dry matter accumulation, annual grain yield, and economic benefits were investigated, in order to provide the theoretical basis for further optimization of N fertilizer management.【Method】From 2020 to 2023, the field experiment was conducted in the typical wheat-maize double cropping system in Huang-Huai-Hai Plain. The following treatments are set up in the experiment: annual N fertilizer application 400 kg·hm^-2of traditional farmer treatment (CK, wheat season 200 kg·hm^-2: 65% basal fertilizer+35% regreening fertilizer, maize season 200 kg·hm^-2: 100% basal fertilizer), 10% reduction of annual N fertilizer (SN, wheat season 180 kg·hm^-2: 50% basal fertilizer+50% regreening fertilizer, maize season 180 kg·hm^-2: 100% basal fertilizer), 20% reduction of annual N fertilizer (NH, wheat season 180 kg·hm^-2: 22.2% basal fertilizer+33.3% regreening fertilizer+44.5% heading fertilizer, maize season 140 kg·hm^-2: 28.6% basal fertilizer+71.4% pre-tasseling fertilizer), and 30% reduction of annual N fertilizer (NL, wheat season 140 kg·hm^-2: 43% regreening fertilizer+57% heading fertilizer, maize season 140 kg·hm^-2: 100% pre-tasseling fertilizer). The photosynthetic physiological characteristic, grain yield, and economic benefits of the wheat-maize double cropping system were tested. 【Result】Under the condition of N reduction, the photosynthetic physiological characteristic of crops were optimized by N fertilizer postponing. Averaged across the three years, NL significantly increased leaf area index by 19.0%-40.1% and 21.6%-36.7% at jointing and anthesis of winter wheat, respectively, compared with CK, SN, and NH treatments. Meanwhile, NL was 6.8%-7.3% higher at silking of summer maize than CK and SN. Similarity, for winter wheat, averaged across the three years, NL significantly increased SPAD value by 7.7%-10.0% and 7.4%-12.9% at jointing and anthesis, respectively, compared with CK and SN. Additionally, it showed a 5.2%-16.2% increase at mid-filling compared with CK, SN, and NH, respectively. Meanwhile NL and NH were 9.0%-9.4% and 6.7%-7.1% higher at pre-tasseling of summer maize, respectively, compared with CK and SN, and 5.1%-9.4% and 4.1%-9.2%at silking and mid-filling under NL treatment, respectively, compared with CK, SN, and NH. Averaged across the three years, NL significantly increased Pn by 8.9%-13.3%, 14.0%-18.1%, and 20.1%-24.4% at jointing, anthesis, and mid-filling of winter wheat, respectively, compared with CK and SN. Meanwhile NL was 4.2%-5.7%, 8.7%-13.4%, and 7.7%-12.8% higher at pre-tasseling, silking, and mid-filling, respectively, compared with CK, SN, and NH, respectively. NL treatment improved the aboveground dry matter accumulation rate and stabilized or increased the aboveground dry matter accumulation at different growth stages by N fertilizer postponing. Average across the three years, the aboveground dry matter under NL significantly increased by 26.7%, 27.4%, and 18.1% at jointing, anthesis, and maturity of winter wheat, respectively, compared with CK. Additionally, there was a 14.4% increase in dry matter accumulation at silking of summer maize under NL compared with CK. N fertilizer postponing improved the photosynthetic physiological characteristics and promoted aboveground dry matter accumulation at each growth stage under the condition of N reduction, and increased winter wheat, summer maize, and annual grain yield and economic benefits. Compared with CK, the results averaged from 2020 to 2023 showed that NL treatment significantly increased the grain yield and economic benefits by 20.5%, 18.1%, 19.1% and 32.4%, 23.8%, 27.9%, respectively.【Conclusion】In the wheat-maize double cropping system on Huang-Huai-Hai Plain, a 30% reduction of annual N fertilizer by N fertilizer postponing could optimize crops photosynthetic physiological characteristics (such as leaf area index, leaf SPAD value, and Pn), as well as enhance aboveground dry matter accumulation rate, and aboveground dry matter accumulation at different growth stages. This would ultimately lead to improved winter wheat, summer maize, and annual grain yield, resulting in enhanced economic benefits.

The Effect of Nitrogen Fertilizer on Nitrogen Use Efficiency and Yield of Foxtail Millet in Ridge-Furrow Rainwater Harvesting Planting Model

ZHOU HaoLu, SHEN ZhaoYang, LUO XinYu, HUANG YingHui, WANG KeXin, WANG YunHao, GAO XiaoLi

Scientia Agricultura Sinica. 2024, 57(5):  885-899.  doi:10.3864/j.issn.0578-1752.2024.05.005

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【Objective】To explore the response of foxtail millet growth and yield to nitrogen fertilizer under ridge-furrow rainwater harvesting planting model, in order to provide a theoretical basis for high yield and efficient production of foxtail millet in the semi-arid regions of the Loess Plateau. 【Method】Shaan-Yugu No. 3 variety was selected for field experiments from 2022 to 2023, and split plot design was adopted. The main plot were different planting models (Ridge-furrow rainwater harvesting planting model, R; Traditional plain planting model, T), and the sub-plot were different nitrogen application levels (0 kg·hm^-2, N0; 60 kg·hm^-2, N1; 135 kg·hm^-2, N2; 210 kg·hm^-2, N3). The net photosynthetic rate and chlorophyll content of flag leaves of foxtail millet, dry matter accumulation and nitrogen accumulation of aboveground population at different growth stages, agronomic traits and yield at maturity stage, and nitrogen utilization rate were measured under different planting models. 【Result】Under the same nitrogen application rate, compared with the traditional plain planting model, the net photosynthetic rate and chlorophyll content of flag leaves under the ridge-furrow rainwater harvesting planting model increased only at the filling stage in 2022 with more rainfall and increased at the heading, flowering, and filling stages in 2023 with less rainfall. The ridge-furrow rainwater harvesting planting model enhanced the photosynthetic capacity of foxtail millet in the middle and later stages of growth and delayed leaf senescence during the two years. The dry matter accumulation and nitrogen accumulation of the aboveground population of foxtail millet increased during the jointing, filling, and maturity stages in 2022, and increased at all growth stages in 2023. There was no significant difference between N1, N2, and N3, and the aboveground dry matter accumulation and nitrogen accumulation increased by 0.6% to 39.5% and 0.9% to 51.1%, respectively. The grain nitrogen accumulation, post-flowering nitrogen accumulation, and post-flowering nitrogen contribution rate to grain have significantly increased over the past two years, meeting the nutrient needs of foxtail millet throughout its entire growth stage and improving insufficient nitrogen supply in the later growth stage of foxtail millet under traditional plain planting model. Nitrogen partial productivity increased between two years, increasing by 3.7% to 14.3% and 2.8% to 27.6%, respectively. Nitrogen agronomic efficiency slightly decreased at a nitrogen application rate of 210 kg·hm^-2 in 2022, while the nitrogen agronomic efficiency and nitrogen use efficiency both decreased when the nitrogen application rate exceeded 135 kg·hm^-2 in 2023. Meanwhile, nitrogen partial productivity, nitrogen agronomic efficiency, and nitrogen use efficiency decreased with the increase in nitrogen application rate. The ear diameter and single ear weight of foxtail millet increased between two years, while the ear length decreased. The yield has significantly increased between two years, increasing by 3.7% to 14.3% and 2.8% to 27.6%, respectively. The yield increase effect was more significant in 2023 with less rainfall, and there was no significant increase in yield when the nitrogen application rate exceeded 60 kg·hm^-2. 【Conclusion】In the semi-arid regions of the Loess Plateau, the ridge-furrow rainwater harvesting planting model combined with 60 to 135 kg·hm^-2 nitrogen fertilizer is more beneficial to the nitrogen utilization and yield formation of foxtail millet.

PLANT PROTECTION

Identification and Gene Function Analysis of StCks1, a Cyclin- Dependent Kinase Subunit of Setosphaeria turcica

ZHANG BoWen, ZHAO LiWen, XU Lu, LI Pan, ZENG FanLi, MENG YaNan, DONG JinGao

Scientia Agricultura Sinica. 2024, 57(5):  900-908.  doi:10.3864/j.issn.0578-1752.2024.05.006

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【Objective】Cks1 is a structural subunit of the cyclin-dependent kinase complex CDK, and is a key gene in the process of cell cycle regulation. The aim of this article is to identify the StCks1 of Setosphaeria turcica, analyze the differences in StCks1 expression and its interacting proteins during the development of appressorium of S. turcica, and to lay the foundation for further research on the role of StCks1 in the development of appressorium.【Method】By analyzing the entire genome data of the wild-type strain 01-23 of S. turcica, the StCks1 protein sequence was obtained. Using software MEGA 5.0 and online databases Pfam and SMART, phylogenetic tree construction and conserved domain analysis of Cks1 proteins from different species such as Saccharomyces cerevisiae, Schizosaccharomyces cerevisiae, and Arabidopsis thaliana were conducted. Samples were collected from different stages of appressorium of S. turcica for transcriptome sequencing and expression analysis. Using a prokaryotic induction expression system, a prokaryotic expression vector pGEX-6p-3-StCks1 was constructed, and transformed the E. coli expression strain BL21 to induce expression and purification of the recombinant protein GST-StCks1. The interaction protein of StCks1 was identified through GST pull-down, Western blot, and yeast two-hybrid assay.【Result】The only StCks1 protein coding gene was identified in the entire genome of S. turcica, which is composed of 130 amino acids. Its tertiary structure mainly consists of four strands of β-fold and three α helices, containing HxPEPH (His-any-Pro-Glu-Pro-His) conserved sites and Cks conserved domains. Through transcriptome sequencing and expression analysis, it was found that the expression level of StCks1 was significantly up-regulated at different stages of appressorium development. A large amount of soluble protein could be obtained by inducing recombinant protein GST-StCks1 at 1 mmol·L^-1 IPTG and 30 ℃ for 2 h. A large number of interacting proteins were obtained using GST pull-down technology and identified by mass spectrometry. Western blot and yeast two-hybrid experiments were conducted to verify the interaction between StCks1 and the cyclin-dependent kinase CDK1.【Conclusion】There is a unique StCks1 in S. turcica, which regulates the development of appressorium through interaction with the cyclin-dependent kinase CDK1.

The Predation Behavior of Odontomachus monticola and Camponotus japonicus on Black Cutworm (Agrotis ipsilon)

GAO DongMei, HUANGFU JiaYi, GUO Xiao

Scientia Agricultura Sinica. 2024, 57(5):  909-918.  doi:10.3864/j.issn.0578-1752.2024.05.007

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【Objective】Black cutworm (Agrotis ipsilon) is one of the important underground pests of food and cash crops. The objective of this study is to clarify the predation characteristics of Odontomachus monticola and Camponotus japonicus on A. ipsilon, enrich the biological control techniques of A. ipsilon, and to reduce the cost of A. ipsilon control and the amount of chemical pesticides.【Method】Predation selectivity, predation mode, predation lethality on different A. ipsilon stages by O. monticola and C. japonicus were observed and counted under two natural conditions, soil cover and bare prey, for 1, 6, 12, 24 and 48 h. Sequence characteristics of predation behavior were carried out indoors using experimental populations of the two ant species. Statistical analyses were performed to clarify the differences in predation behavior and pest control characteristics of the two ant species.【Result】C. japonicus was less lethal to A. ipsilon eggs, and the cumulative 48 h lethality was only 12.67%. O. monticola did not feed on A. ipsilon eggs. The control effect of the two ant species on A. ipsilon larvae and pupae was over 49% and up to 98.67%, and the control effect decreased with the age of A. ipsilon. There were some differences in the sequence characteristics of predation behavior of O. monticola and C. japonicus: for A. ipsilon low instar larvae, both ants mainly carried directly; for the larger A. ipsilon elder instar larvae, C. japonicus had behaviors of probing, attacking, calling companions, cutting prey and carrying in groups, which showed obvious group behavior. On the other hand, the predation behavior of O. monticola on large prey was simple, and behaviors such as calling companions and group handling were not observed, the group predation behavior was not obvious. The differences in the predation behavior of the two ant species on A. ipsilon were also reflected in the mode of predation and lethality on A. ipsilon: Most of the low instar A. ipsilon larvae were carried directly by C. japonicus, the predation efficiency was very high, and C. japonicas usually preyed A. ipsilon elder instar larvae in groups. O. monticola had a higher proportion of discarded elder A. ipsilon larvae, but A. ipsilon larvae that escaped predation by biting and stinging had a higher mortality. In addition, soil cover strongly influenced the detection of A. ipsilon larvae by ants, the two ant species could only detect surface prey and not subterranean prey.【Conclusion】C. japonicus and O. monticola have better control effect on both A. ipsilon larvae and pupae, and the lower the age of A. ipsilon, the better the control effect. There are some differences in the predation behavior of the two ant species on the elder A. ipsilon larvae, but it does not affect the control effect on A. ipsilon.

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

Effects of Localized Nitrogen Supply on Plant Growth and Water and Nitrogen Use Efficiencies of Maize Seedling Under Drought Stress

WANG Yu, ZHANG YuPeng, ZHU GuanYa, LIAO HangXi, HOU WenFeng, GAO Qiang, WANG Yin

Scientia Agricultura Sinica. 2024, 57(5):  919-934.  doi:10.3864/j.issn.0578-1752.2024.05.008

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【Objective】Frequent spring drought and excessive application of nitrogen (N) fertilizer would hinder maize seedling growth and development, and may negatively affect plant growth during middle-late periods and final grain yield. This study was conducted to determine the effects of localized supply of N fertilizer on maize seedling growth, root morphology, water and N utilization under drought stress conditions, ait of nitrogen inputs on graming to provide a technical basis for promoting root development, improving water and N use efficiencies, and achieving high and stable yields in maize production.【Method】The split-root pot experiments with different water and N levels were conducted in 2021 and 2022. Five N supply patterns were designed: no N supply (N0/N0), uniform low N supply (LN/LN), localized low N supply (LN/N0), uniform high N supply (HN/HN), and localized high N supply (HN/N0). The N rates were 0.12 and 0.24 g N·kg^-1 dry soil for LN and HN levels, respectively. Water management was initiated from 3-leaf stage and lasted for 3 weeks, including three soil moisture content: severe water-stress (35% of field capacity, W0), moderate water-stress (55% of field capacity, W1), and well-watered conditions (75% of field capacity, W2). At the end of the water management period, plant growth traits, shoot dry matter (DM), shoot N uptake, root DM, root length, and water and N use efficiency (WUE, NUE) were measured. 【Result】 The water stress significantly limited plant growth, DM accumulation, and N uptake in maize seedlings, but increased root shoot ratio. Compared with W2, the total root length under W0 was reduced by 48.0%, while the effects were relatively less under W1. The water stress reduced NUE in maize, with average decreases of 10.1 and 4.6 percentage points under W0 and W1, respectively, compared to W2, and the WUE was decreased by 19.4% under W0 and increased by 11.9% under W1. The N supply patterns also significantly affected plant growth, DM accumulation, N uptake and utilization in maize seedlings, and the two factors of water and nitrogen showed significant interaction. Based on the results obtained in 2022, the highest shoot DM was observed in LN/LN, and which were 8.3%, 12.6% and 23.6% higher than those in N0/N0 under W0, W1 and W2, respectively. Nevertheless, the highest root DM was observed in LN/N0, with the increases of 9.5%, 17.0%, and 31.2% higher than that in N0/N0 under the three water levels. Moreover, the LN/N0 increased significantly root﹕shoot ratio under both W1 and W2. The HN/HN had the most severe negative impacts on maize seedling growth, with 30.1%, 14.6%, and 7.0% lower shoot DM as compared with N0/N0 under W0, W1 and W2, respectively. The larger decreases in root DM (41.0%, 44.2% and 34.9%, respectively) were observed in HN/HN, thus resulting in significant reductions in root﹕shoot ratio. The HN/N0 showed a relatively less effect on shoot DM, but significantly reduced root DM and root﹕shoot ratio. Compared with N0/N0, both HN/HN and HN/N0 resulted in significant decreases in root length, whereas LN/N0 significantly increased total root length by promoting root proliferation on the side without N supply. The application of N fertilizer significantly increased plant N uptake, with higher value in LN/LN but lower in LN/N0 across all water levels. With regard to NUE, which was higher in LN/LN while lower in HN/HN. The differences in N uptake and NUE among treatments increased with reducing water stress levels. Both plant water consumption and WUE were higher in LN/LN and LN/N0, followed by HN/N0, whereas the lowest values were observed in HN/HN. Compared with N0/N0, uniform or localized low N supply showed promoting effects on maize seedling growth, while uniform or localized high N supply had negative effects that intensified with increasing water stress levels. Overall, localized N supply had a greater effect on root growth relative to uniform N supply, inducing a morphological plasticity response in the root system. The correlation analysis results indicated that shoot DM, WUE and NUE had positive and significant relationships with total root length of maize seedlings across different water levels and N supply patterns. For the localized N supply treatments, the correlation between root length on the side without N supply and shoot DM, WUE and NUE was higher.【Conclusion】In comparison to uniform N supply, the localized N supply effectively promoted root proliferation on the side without N supply during maize seedling stage, resulting in increased root DM and total root length. Hence, the localized supply with a lower N rate was recommended to enhance drought tolerance of maize seedlings and improve WUE.

Effects of Biochar and Straw Return on Greenhouse Gas Emissions and Global Warming Potential in the Farmland

GAO ShangJie, LIU XingRen, LI YingChun, LIU XiaoWan

Scientia Agricultura Sinica. 2024, 57(5):  935-949.  doi:10.3864/j.issn.0578-1752.2024.05.009

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【Objective】The global warming potential of agro-ecosystem is an important part of affecting global climate change. This study clarified the effects of biochar and straw application on the global warming potential of farmland through field experiments, in order to provide theoretical basis for mitigating climate change and agricultural waste resource utilization. 【Method】In our study, a three-year field positioning experiment was carried out in the winter wheat-summer maize rotation farmland in the Agro-ecosystem Experimental Station in Huantai County, Shandong Province. Four treatments were set up in the experiment: ① control (CK); ② biochar (C); ③ straw return(S); ④ straw return combined with biochar (CS). The nitrogen, phosphorus and potassium fertilizers were applied in all treatments. The nitrogen fertilizer was 200 kg·hm^-2·a^-1 urea, the phosphate fertilizer was 55 kg·hm^-2·a^-1 superphosphate and the potassium fertilizer was 40 kg·hm^-2·a^-1 potassium sulfate. We measured the flux of greenhouse gases (CO₂, N₂O and CH₄) by static chamber-gas chromatography method, calculated the net global warming potential (NGWP) and greenhouse gas emission intensity (GHGI), and analyzed the effect of biochar and straw on the greenhouse gas emissions and the net global warming potential. 【Result】(1) From the emission of greenhouse gases in the three-year experiment, the average annual cumulative Re emissions of S and CS treatments increased by 47.8% and 67.9% (P<0.05), respectively, compared with CK. The average annual cumulative N₂O emission under C treatment reduced by 20.3% than CK treatment (P<0.05), and the cumulative N₂O emission under S and CS treatments increased by 23.6% and 41.4% (P<0.05), respectively. (2) In comparison with CK treatment, soil organic carbon change (ΔSOC) of C, S and CS treatments increased significantly, and the largest increase was in the CS treatment by 150.6% (P<0.05). Compared with the first year, soil organic carbon change (ΔSOC) of C, S and CS treatments increased by 21.7%, 20.8% and 17.8% (P<0.05), respectively. There were significant differences in net global warming potential (NGWP) and greenhouse gas emission intensity (GHGI) among all treatments. Compared with CK treatment, the average annual NGWP of C, S and CS treatments decreased by 163.5%, 171.7% and 273.0% (P<0.05). Compared with the first year, the NGWP of C, S and CS treatments in the third year decreased by 73.4%, 58.8% and 54.7% (P<0.05), respectively. The annual average GHGI of C, S and CS treatments decreased by 236.2%, 253.3% and 388.9% than CK treatment (P<0.05), respectively. In comparison with the first rotation, the GHGI of C, S and CS treatments in the third rotation decreased significantly by 126.3%, 98.2% and 108.6% (P<0.05), respectively. The yield of C, S and CS treatments remained stable with a slight increase, but there was no significant difference between the yield of CK, C, S and CS treatments. 【Conclusion】Compared with applying chemical fertilizer alone, the methods of adding biochar, straw return and straw return combined with biochar could prevent the NGWP without reducing crop yield. Among these treatments, straw return combined with biochar could reduce the net global warming potential to the greatest extent. Therefore, straw return combined with biochar is an effective measure to enhance carbon sequestration and mitigate climate change.

Spatial Distribution and Driving Factors of Winter Wheat Irrigation Carbon Emission Intensity at Township Level in Henan Province

ZHU RuiMing, ZHAO RongQin, JIAO ShiXing, LI XiaoJian, XIAO LianGang, XIE ZhiXiang, YANG QingLin, WANG Shuai, ZHANG HuiFang

Scientia Agricultura Sinica. 2024, 57(5):  950-964.  doi:10.3864/j.issn.0578-1752.2024.05.010

Abstract ( 67 )   HTML ( 6 )   PDF (2852KB) ( 55 )   Save

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【Objective】The aim of this study was to explore the spatial distribution pattern and driving mechanism of agricultural carbon emissions of different townships and irrigation modes, and to reveal the regional differences of agricultural water and energy resources consumption combination characteristics, so as to provide necessary references for agricultural irrigation mode optimization, water and energy resources conservation and low-carbon agricultural sustainable development. 【Method】The irrigation carbon emission intensity and its spatial distribution of winter wheat at township level were calculated by using multi-source data, including remote sensing data, statistical data, meteorological data, and agricultural irrigation survey data. In addition, the geographical detector was employed to discuss the influencing factors of carbon emission intensity under different irrigation modes (Irrigation mode with rain-fed agriculture as the primary method, PI mode; Irrigation mode with surface water irrigation as the primary method, SWI mode; Irrigation mode with diversion and irrigation projects as the primary method, WDI mode; Irrigation mode with groundwater irrigation as the primary method, GI mode). 【Result】(1) In 2018, the average carbon emission intensity of winter wheat irrigation at the township level in Henan Province was 15.05 kg∙t^-1, which showed a west-to-east decreasing trend in the longitudinal direction and a north-high-south-low spatial distribution pattern in the latitudinal direction. The mean value of mean carbon emission intensity at township level was 15.05 kg∙t^-1. The high value townships was concentrated in the western mountainous region, while the lower gathered in the southeast region. (2) By comparing four irrigation modes, the irrigation mode with highest carbon emission intensity was the irrigation mode with rain-fed as the primary method (PI mode, 22.22 kg∙t^-1), while the irrigation mode with groundwater irrigation as the primary method was the least ideal (SWI mode, 11.05 kg∙t^-1). (3) The average elevation and effective precipitation explained 49% and 39% of the spatial differentiation pattern of carbon emission intensity, respectively, which were key factors influencing the carbon emission intensity of winter wheat irrigation. The effect of the same driving factor under different irrigation modes was variable. The influence of energy consumption on the spatial differentiation pattern of winter wheat irrigation carbon emission intensity under the irrigation mode dominated by water diversion irrigation projects (WDI mode) was the strongest, and the driving effect of land input intensity under the GI mode was the most significant. In addition, the interaction of different drivers had double-factor enhancement effects and non-linear enhancement effects. 【Conclusion】There was spatial heterogeneity and differences in irrigation mode in the carbon emission intensity of winter wheat irrigation at the township level in Henan Province. The PI mode was the mode with the highest winter wheat irrigation carbon emission intensity and the WDI mode was the mode with the highest carbon emissions. The average elevation and effective precipitation were the main reasons for the formation of spatially divergent patterns of irrigation carbon emission intensity of winter wheat at township scale in Henan Province, while anthropogenic factors were the dominant forces under different irrigation patterns.

HORTICULTURE

Light Interception Rate and Canopy Structure Optimization of Korla Fragrant Pear

YAN Pan, WANG ZhenDong, DENG YongHui, CHEN QiLing, ZHENG QiangQing

Scientia Agricultura Sinica. 2024, 57(5):  965-979.  doi:10.3864/j.issn.0578-1752.2024.05.011

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【Objective】 This study investigated the canopy structure of five tree shapes of Korla fragrant pears, analyzed the differences in light distribution within different tree shapes, established a daily change model of light interception, calculated the light interception rate, clarified the regulation methods and target parameters of the canopy structure, to provide a reference basis for cultivating high-light-efficiency tree shapes. 【Method】 Korla fragrant pear trees with different shapes were chosen as the test materials, the canopy structural parameters and photosynthetic-effective-radiation (PAR) passing through different tree shapes were measured. Based on a quadrant approach, a three-dimensional space was constructed, the light distribution maps were drawn, an evaluation index system of light interception was established, and the light interception amount and light interception rate were calculated. Through correlation analysis and principal component analysis, the main canopy structure and regulation methods that affect light interception were identified. 【Result】 (1) The ground diameter and average branch diameter of large cylindrical-shaped trees were larger, with main branches with imbalanced branch-to-trunk ratios. The number of branches and branch diameter through the canopy of the cylindrical-shaped trees were more uniform. The number of branches, total length of branches, and average branch length of the narrow cylindrical-shaped trees were significantly lower than those of the cylindrical-shaped trees. The height of trees with the short cylindrical-shaped trees was significantly lower and other parameters were similar to the narrow cylindrical-shaped trees. The ground diameter, average branch length, average branch diameter, average branch angle, and average distance of branches of slender- spindle-shaped trees were significantly lower than those of cylindrical-shaped trees, but the proportion of short branches was significantly larger. (2) The distance from the trunk and height were the main factors which affected the canopy light interception (LI), and the average light interception(ALI) at a distance of 100 cm from the trunk increased significantly, reached 572 μmol·m^-2·s^-1, which was approximately twice that of the inner chamber. The lighting conditions below 220 cm were poor. The south, north, southeast, and southwest sides were high-light areas, while the west, east, northeast, and northwest sides were low-light areas. The daily variation in ALI can be roughly divided into five periods. (3)As the tree shape became narrower and shorter, the ALI significantly increased in the inner chamber, in each or partial layer, in all or partial directions, and in partial periods. (4) The single-day cumulative light interception (CLI) of slender-spindle-shaped trees was 22.2 mol·m^-2, the group CLI was 3 712 mol/667 m², and the light interception rate (LIR) was 35.6%, which was significantly higher than that of other tree shapes. The proportion of low-light area (PLL) was 50.9%, which was significantly lower than that of other tree shapes. (5) Five canopy structural parameters were significantly positively correlated with the LIR while fifteen canopy structural parameters were significantly negatively correlated. 【Conclusion】 The proportion of short branches is the most important parameter affecting the light interception in Korla fragrant pear trees, while the length of branches is the most important parameter affecting the proportion of low-light area. Control the tree height and canopy width can improve the light interception rate and the uniformity of light distribution. The slender-spindle-shaped canopy has a more uniform light distribution and the highest light interception rate. It can maintain a larger number of branches and a larger proportion of short branches with reduced average branch length to improve the lighting level of the canopy.

Isolation and Functional Verification of Genes Mediating Mineral Element Stress Tolerance in Lotus

HU HengLiang, GU TianYu, CHEN SiYing, WANG Yao, PENG JiaShi

Scientia Agricultura Sinica. 2024, 57(5):  980-988.  doi:10.3864/j.issn.0578-1752.2024.05.012

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【Objective】 Lotus (Nelumbo nucifera) is a traditional edible and medicinal homologous crop in China, yet its abundant genetic resources lack functional characterization. This research aimed to functionally identify genes involving in the accumulation/tolerance of trace mineral nutrients and heavy metal elements in lotus, thereby accumulated gene resources that facilitated increased nutrient efficiency and stress tolerance, so as to provide a theoretical basis for the genetic improvement of lotus. 【Method】The representative Xianglian lotus variety, Cunsanlian was used as the experimental material. Initially, a yeast expression cDNA library of lotus was constructed and transformed into Saccharomyces cerevisiae. It was then screened on plates containing excessive cadmium (Cd), manganese (Mn), zinc (Zn), copper (Cu), iron (Fe), and aluminum (Al) stress to isolate target genes mediating stress tolerance in positive yeast clones. Finally, the functional verification of the selected tolerance genes was conducted through combining bioinformatics analysis and yeast complementation verification. 【Result】A yeast cDNA library of lotus was constructed with a capacity exceeding 10⁶ yeast monoclonal clones, a recombination rate of 100%, and an average insertion fragment size greater than 1 000 bp. Following the screening on stress plates, 13 genes for Cd tolerance, 4 for Mn tolerance, 4 for Zn tolerance, 3 for Cu tolerance, 7 for Fe tolerance, and 1 for Al tolerance were identified. Among these, 3 genes were able to mediate tolerance to both Fe and Mn. These genes were distributed on all lotus chromosomes except chromosome 6.【Conclusion】A high-quality yeast expression cDNA library of lotus was constructed, and screened 29 genes mediating resistance to excess trace mineral nutrients or heavy metal elements, accumulating gene resources for lotus genetic improvement for enhanced nutrient efficiency and the prevention of heavy metal accumulation.

FOOD SCIENCE AND ENGINEERING

Effects of Novel Phase Change Coolant on the Postharvest Quality of Shiitake Mushrooms

HUANG Hao, WU QingHong, ZHANG Yu, WANG Ye, LIU QingE, FANG YiDa, LUO ZiSheng

Scientia Agricultura Sinica. 2024, 57(5):  989-999.  doi:10.3864/j.issn.0578-1752.2024.05.013

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【Background】Edible mushrooms with unique flavor are rich in protein, vitamins, and a variety of bioactive components, such as polysaccharides, phytosterol, etc. China is the major country in the cultivation and consumption of edible mushrooms, among which shiitake mushroom (Lentinula edodes) is the most widely cultivated. The current coolants exhibit low phase change temperature, which may result in chilling injury of shiitake mushrooms.【Objective】 Based on the freezing temperature of shiitake mushrooms, a novel phase change coolant specially for shiitake mushrooms was developed to maintain the postharvest quality.【Method】The main component of the coolant was screened by differential scanning calorimetry (DSC) based on the phase change latent heat and phase change temperature. Potassium sulfate, nano titanium dioxide, and superabsorbent resin were mixed to prepare the coolant. The effects of coolant on the qualities of shiitake mushrooms were studied by the weight loss rate, color, hardness and other indexes. The activities of enzymes related to glutamate metabolism and energy metabolism were also measured to explore the mechanism of preservation.【Result】 The aqueous solution of maltitol was selected as the main component by DSC. The optimal compositions of coolant were 1.85% of maltitol, 2.35% of potassium sulfate, 0.02% of nano titanium dioxide, and 0.80% of super absorbent resin, leading to the phase change latent heat and temperature at 405.26 J·g^-1 and -1.8 ℃, respectively. Compared with the control group, under the low-temperature conditions provided by the refrigerant, the weight loss rate of shiitake mushrooms decreased by 51.92%, and the hardness and brightness increased by 66.67% and 41.94%, respectively. The shelf life of shiitake mushrooms was extended. The low temperature also regulated the activities of enzymes, which were related to glutamate metabolism and energy metabolism, the glutamic acid and energy charge level increased by 36.64% and 54.76%, respectively. Finally the freshness of shiitake mushroom was maintained.【Conclusion】The novel phase change coolant based on the freezing temperature of shiitake mushrooms could maintain the weight, color, hardness and other indexes, regulate the level of glutamic acid and energy, and delay the deterioration of shiitake mushrooms. Therefore, the coolant was effective for the cold-chain preservation of shiitake mushrooms.

ANIMAL SCIENCE·VETERINARY SCIENCE

miR-34a Induces Early Apoptosis of Porcine Ovarian Granulosa Cells by Activating lncRNA NORHA Transcription

WANG SiQi, ZHOU ChunXue, LI YuQi, DU Xing, PAN ZengXiang, LI QiFa

Scientia Agricultura Sinica. 2024, 57(5):  1000-1009.  doi:10.3864/j.issn.0578-1752.2024.05.014

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【Objective】 miR-34a has been shown to be a pro-apoptotic factor in porcine ovarian granulosa cells (GCs) in our previous study. The aim of this study was to explore the RNA activation mechanism by which miR-34a induced apoptosis in the nucleus, so as to provide a basis for uncovering the molecular mechanism of porcine follicular atresia, and to screen small nucleic acid regulator that regulated procine reproduction. 【Method】 Bioinformatics methods were used to analyze the characteristics of the porcine miR-34a, the binding sites of miR-34a to the promoter region of target lncRNA NORHA, and the binding capacity. The subcellular localization of miR-34a was analyzed in porcine GCs by using nucleocytoplasmic fractionation. The effects of miR-34a on the expression of NORHA, apoptosis marker genes BCL-2 and BAX in porcine GCs were determined by qPCR. Luciferase assay was used to verify the regulatory relationship of miR-34a on the transcriptional activity of the NORHA promoter. Chromatin immunoprecipitation (ChIP) was utilized to detect the enrichment of AGO2 and histone modifiers at the miRNA response element (MRE) in the NORHA promoter in porcine GCs by miR-34a. The regulatory effect of miR-34a on downstream factor FOXO1 expression and GC apoptosis via NORHA were analyzed by western blot and flow cytometry. 【Result】 The porcine miR-34a was an intergenic miRNA, and its sequence was highly conserved among mammals. Subcellular localization analysis showed that miR-34a was distributed in both nucleus and cytoplasm in porcine GCs. Bioinformatic analysis revealed that the MRE of miR-34a was located at -194 nt - -173 nt in the NORHA promoter, and the binding free energy of the two compounds was -23.9 kcal·mol^-1. qPCR showed that miR-34a significantly upregulated NORHA expression in porcine GCs. Luciferase assay revealed that miR-34a significantly upregulated the activity of the reporter vector with NORHA promoter, whereas it had no significant effect on the activity of promoter with the mutated MRE of miR-34a. ChIP assay showed that miR-34a increased the enrichment of AGO2, a core member of RNA induced transcriptional activation (RITA) complex, and H3K4me3, a transcription activated histone modifier, while or decreased the enrichment of H3K9me3, a transcription repressed histone modifier at MRE motif of miR-34a in the NORHA promoter. Co-transfection assay showed that knockdown of NORHA significantly reversed the upregulation of FOXO1 protein level and early apoptosis rate caused by miR-34a overexpression, as well as the downregulation of the BCL-2/BAX ratio. 【Conclusion】 Nuclear miR-34a was an endogenous small activating RNA (saRNA) in porcine GCs, and might be involved in the initiation of porcine follicular atresia by inducing early apoptosis in porcine GCs through target activating the transcription of the pro-apoptotic lncRNA NORHA.

The Potential and Mechanisms of Apigenin to Relieve Heat Stress and Hypoxia in Dairy Cows Based on Network Pharmacology and Molecular Docking

LIU ZhuoLin, LIU HongYun

Scientia Agricultura Sinica. 2024, 57(5):  1010-1022.  doi:10.3864/j.issn.0578-1752.2024.05.015

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【Objective】 The study aimed to predict the mechanisms of apigenin in relieving heat stress and hypoxia stress in dairy cows by using network pharmacology and molecular docking so as to provide a reference for the application of apigenin.【Method】 Firstly, the targets related to apigenin, heat stress, and hypoxia stress were obtained from TCMSP and GeneCards databases, and apigenin targets in relieving dual stress were obtained by Venn intersection. The protein-protein interaction (PPI) network was constructed using the STRING database, and the core targets were identified by analyzing the network node centrality using the CytoNCA plugin in Cytoscape 3.9.1 software. Autodock vina program was used to perform molecular docking of apigenin with the identified targets. Perform gene ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the top 30 ranked targets based on degree centrality using Functional Annotation, the analysis tool of David database. Additionally, Molecular complex detection analysis (MCODE) of the above targets yielded the core gene clusters, then GO and KEGG analysis were performed. Apigenin-dual stress targets-signaling pathway network model were constructed through the KEGG pathway enrichment results and core target information. Finally, apigenin was docked to the core targets using the Autodock vina program, and the results were visualized using the Pymol software and the PLIP website.【Result】 The TCMSP database retrieved 68 targets of apigenin, 6 661 hypoxia-related genes and 9 046 heat stress-related genes were obtained in GeneCards database, and 56 apigenin targets in relieving dual stress were obtained. The number of nodes in the target PPI network was 56, the number of edges was 436, the average node degree value was 15.6, the mean local clustering coefficient was 0.728, and PPI enrichment P value was less than 1.0×10^-16. The core targets were AKT1 (RAC-alpha serine/ threonine-protein kinase), TP53 (Cellular tumor antigen p53), TNF (Tumor necrosis factor), CASP3 (Caspase-3), INS (Insulin), BCL2L1 (Bcl-2-like protein 1), VEGFA (Vascular endothelial growth factor A), HIF1A (Hypoxia-inducible factor 1-alpha), PTGS2 (Prostaglandin G/H synthase 2), and SREPINE1 (Plasminogen activator inhibitor 1). The molecular docking results showed that apigenin could be stably bound to the above core targets. A total of 54 Biological Processes (BP), 6 Molecular Functions (MF), and 11 Cellular Components (CC) were obtained by GO functional enrichment analysis (P<0.01), and KEGG were enriched to 98 signaling pathways. MCODE analysis showed two major gene clusters. 【Conclusion】 Apigenin mainly regulated cell proliferation, apoptosis, oxygen sensing and inflammation through PI3K-AKT, p53, HIF-1, NF-kappa B and other pathways, to further alleviate heat stress and hypoxia stress. The study provided a theoretical reference for the application of apigenin and apigenin-rich plants in dairy cow production.