Table of Content

20 October 2025, Volume 34 Issue 10

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The impact of shrub encroachment on species niches and interspecific associations of the Artemisia scoparia community in desert grassland

Ze-hua LIU, Lin CHEN, Ya-qi ZHANG, Jin-xiao LONG, Xue-bin LI, Dan-bo PANG

2025, 34(10):  1-15.  DOI: 10.11686/cyxb2024477

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The impact of shrub encroachment on species niches and interspecific associations within the Artemisia scoparia community was investigated through vegetation surveys conducted in typical Ammopiptanthus mongolicus shrub-encroached sites in desert grasslands. Species niches and interspecific associations within the A. scoparia community were analyzed using niche width， niche overlap， detrended correspondence analysis （DCA）， variance ratio， chi-square testing， association coefficient （AC）， Ochiai （OI） and Spearman rank correlation methods. The results reveal an A. scoparia community consisting of 17 herbaceous species and 3 semi-shrub species， predominantly from the Poaceae and Fabaceae families， highlighting the complexity of the community composition. The importance value and niche width of the A. scoparia community exhibited significant changes with both parameters gradually increasing as the degree of shrub encroachment decreased. In plots with moderate shrub cover， A. scoparia displays a high niche overlap with most species， particularly with Salsola collina （0.87）， suggesting a strong similarity in their resource requirements. The DCA ordination results indicate clear clustering of species distributions in the plots， while the species distribution in less encroached plots is relatively more dispersed. No significant positive or negative associations were observed in the communities of heavy shrub， moderate shrub， light shrub， and shrub-free grasslands. The results of the chi-square testing， AC， OI and Spearman rank correlation analysis were consistent， reinforcing the overall pattern of weak species associations， low independence and an immature community structure. In conclusion， shrub encroachment alters the species niches and interspecific associations within the A. scoparia community in desert steppe ecosystems， contributing to the instability of the community. During ecosystem restoration efforts， it is recommended to select species combinations that exhibit positive associations with dominant species such as A. scoparia， thereby fostering a more stable community structure. This approach will enhance the ecological stability and recovery of the grassland.

An investigation into the health of synthetic grassland ecosystems in high-alpine regions under varied fertilization and mixed planting circumstances recovering from coal mining damage

Ting LI, Xin-guang YANG, Cheng-wei DUAN, Hua-fang SUN, Tao GAO, Tong-de CHEN, Kai YANG, Qian-hui YANG

2025, 34(10):  16-29.  DOI: 10.11686/cyxb2024455

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The restoration of ecosystems in damaged alpine coal mining areas is a global problem. It is important to ensure the stability of ecosystems in mining areas by optimizing restoration measures. In this study， three sowing methods were implemented， namely： i） revegetation by sowing four gramineous species （Elymus breviaristatus， Poa crymophila， Poa pratensis， Festuca sinensis； gramineous grasslands， designated H）， ii） sowing three Fabaceae species （Medicago ruthenica， Melilotus officinalis， Medicago sativa； leguminous grassland， designated B）， and iii） sowing four Poaceae and three Fabaceae species （gramineous and leguminous grasslands， designated HB）. Additionally， utilizing different proportions of sheep manure and slag soil， four fertilization levels were implemented， namely （sheep manure ： slag soil） heavy fertilization （1∶5， F₁）， medium fertilization （1∶8， F₂）， light fertilization （1∶11， F₃）， and no fertilization （F₄）. Hence， 12 different restoration treatments were generated by factorial combination of these three sowing and four fertilizing methods. The variation in herbage growth and soil properties under the different restoration treatments was evaluated and compared over a number of years. The ecosystem health evaluation was quantitatively judged by using the grassland ecosystem health assessment model OCVOR. The results showed that vegetation cover and aboveground biomass were optimized by the mixed sowing of Poaceae and Fabaceae under the heavy and medium fertilization levels （HBF₁， HBF₂） and increased with time from establishment. Levels of soil organic carbon， available nitrogen， available phosphorus， and available potassium were all substantially higher on the restoration plots after two years of restoration than after one year， but after three years had declined again to levels similar to those measured after one year. In the third year of restoration， the levels of soil organic carbon and available nitrogen were progressively increased （P<0.05） with increased fertilization level， while the law of changes in available phosphorus and available potassium were not significant. The OCVOR index of gramineous grassland under the heavy， medium， and light fertilization （HF₁， HF₂， HF₃）， and also of the leguminous grassland under the heavy fertilization （BF₁） increased significantly （P<0.05） with the years of restoration. Additionally， the OCVOR index of gramineous and leguminous grassland increased significantly under the heavy， medium and light fertilization （HBF₁， HBF₂， HBF₃） （P<0.01） over time. Except for the treatment BF₁， the ecosystem health status and recovery under the treatments HF₁， HF₂， HF₃， HBF₁， HBF₂ and HBF₃ were determined to be unhealthy. Corresponding to these evaluation results， the OCVOR index under the same six treatments increased by 28.61%， 33.03%， 11.46%， 38.96%， 33.79% and 9.83%， respectively， after 3 years of restoration， compared to the first year of restoration. Moreover， the biggest increases were seen under the HBF₁ treatment. Overall， the treatment comprising mixed sowing of Poaceae and Fabaceae under the heavy fertilization level （HBF₁） was found to be most conducive to the health and stability of the ecosystem in the Muli mining area.

Application of desulphurization gypsum with fly ash improves saline-alkali soils

Yu-long YAN, Xue-jun DU, Yuan-yue WANG, Jian-li LIU, Yin-gui DING, Yuan-song WEI

2025, 34(10):  30-40.  DOI: 10.11686/cyxb2024448

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Saline-alkali soil is prevalent in arid and semiarid areas of Dingbian county. The aim of this study was to determine whether adding desulfurization gypsum， fly ash， or a combination of these two materials can improve saline-alkali soil. Pot experiments were conducted with alfalfa （Medicago sativa） growing in saline-alkaline soil with 10 treatments， including a control （CK）， soil with the addition of desulfurization gypsum （1%， 2%， 4%； D₁， D₂， D₃）， soil with the addition of fly ash （1%， 2%， 4%； F₁， F₂， F₃）， and soil with the addition of mixture of desulfurization gypsum and fly ash （0.5%+0.5%， 1%+1%， 2%+2%； DF₁， DF₂， DF₃）. The salinity， sodicity， and nutrient profile of the soils and plant growth were analyzed. The results showed that the addition of desulfurization gypsum reduced the soil pH， increased the total phosphorus content in soil， and increased plant height and biomass. The contents of SO₄^2-， K⁺， Ca²⁺， and Mg²⁺ in soil increased with the addition of desulfurization gypsum. The addition of fly ash increased soil pH， SO₄^2?-， Ca²⁺， and total iron contents， decreased the soil K⁺ content， and resulted in decreased plant height and biomass. The mixture of desulfurization gypsum and fly ash significantly reduced the total iron content and increased the total P content in soil， but did not significantly affect plant growth. The results indicate that， compared with the addition of desulfurization gypsum and fly ash to saline-alkali soil， the addition of desulfurization gypsum alone more effectively improved soil quality and promoted plant growth. These results provide a scientific basis for the improvement of saline-alkali soil and for the comprehensive utilization of coal-based solid waste in arid and semiarid areas.

Response of alfalfa hay yield and nutritional quality to altitude in the Yili River basin

Jin-feng HUI, Kong-qin WEI, Yan-liang SUN, Chun-hui MA, Qian-bing ZHANG

2025, 34(10):  41-50.  DOI: 10.11686/cyxb2024454

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To investigate the effects of altitude on the yield and nutritional quality of alfalfa （Medicago sativa）， we planted four alfalfa varieties in 12 representative plots in the altitude ranges of 1300-1500 m （H₁）， 900-1100 m （H₂） and 500-700 m （H₃） in the Yili River basin， and monitored their growth. Four alfalfa varieties were included in this experiment， namely Algonquin （A₁）， Magnum Ⅶ（A₂）， WL363HQ （A₃）， and Xinmu 4 （A₄）. After harvest， the alfalfa plants were analyzed to determine the contents of crude protein （CP）， neutral detergent fiber （NDF）， and acid detergent fiber （ADF）， the relative feeding value （RFV）， ether extract （EE）， crude ash （Ash）， total phosphorus content （TP） and soluble sugars content （SSC）， and the field hay yield （HY）. The relationship between HY and nutritional indexes of alfalfa was analyzed， and the altitude and varieties suitable for high quality alfalfa feed production were selected based on the results of principal component analysis and membership function analysis. The results show that the CP， EE， RFV， Ash， and HY of alfalfa gradually increased as the altitude decreased， whereas NDF， ADF， and SSC gradually decreased as the altitude decreased. At the same altitude， the RFV， TP， SSC， and HY were significantly higher for A₃ than for the other three alfalfa varieties， and the NDF and ADF contents were lowest in A₃. Pearson’s correlation analyses showed that the HY of alfalfa was positively correlated with CP， RFV， and TP （P<0.05）， but negatively correlated with NDF and ADF （P<0.01）. The CP was positively correlated with TP， EE， RFV， and SSC （P<0.05）. Based on the results of the principal component and membership function analyses， the best performance was observed in A₃ grown at 500-700 m， followed by A₃ grown at 1300-1500 m， and then A₃ grown at 900-1100 m. In conclusion， the yield and nutritional quality of alfalfa plants grown at lower altitudes were superior to those of alfalfa plants grown at higher altitudes in the Yili River basin. Alfalfa variety WL363HQ， coded A₃ in this research，_{can be recommended as suitable for planting at various altitudes in the Yili River basin.}

Quantitative analysis of nitrogen transfer pathways in intercropping systems between alfalfa and Korla pear

Yan MING, Zi-yi DOU, Wei ZHENG, Ning-xin WANG, Xue CHEN

2025, 34(10):  51-61.  DOI: 10.11686/cyxb2024469

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The husbandry methodology of increasing nitrogen supply to Korla pear （Pyrus sinkiangensis） by planting legume forage between rows of Korla pear has been well established， but the specific pathways of nitrogen utilization and transfer under such intercropping are not clear. This study， focused on alfalfa （Medicago sativa） and Korla fragrant pear intercropping. Four nitrogen transfer pathways were established： complete root isolation （PS）， inorganic nitrogen ion transfer only （NS）， hyphal channel transfer （J）， and no isolation （S）. The ¹⁵N isotope tracing method was used to quantify the biological nitrogen fixation capacity， nitrogen transfer effciency， and influencing factors in the intercropping system， as well as to analyze the contributions of each pathway. The results showed： 1） The nitrogen fixation of intercropped alfalfa was 4.74 g·m^-2， which was 0.18 g·m^-2， greater than mono-cropped alfalfa. 2） Compared with PS， the amount of nitrogen fixation increased under S increased by 33.6%， increased under NS by 5.4% and decreased under J by 71.7%， while the amount of nitrogen transfer increased by 224.8%， 119.8% and 4.7%， respectively. The nitrogen transfer efficiencies under the four pathways ranked S （0.92%）>NS （0.63%）>J （0.30%）>PS （0.28%）. 3） Nitrogen fixing enzyme activity had a highly significant effect on the amount of nitrogen fixation by the leguminous intercrop （P<0.01）， while soil hyphal density significantly positively effect on the nitrogen fixation rate （P<0.05） and the intensity and frequency of AM fungal infestation was significantly positively correlated with the rate and amount of nitrogen transfer （P<0.05）. Therefore， the nitrogen transfer pathway under intercropping of alfalfa and pear was mainly a dual pathway of inorganic nitrogen ions and AM fungal mycelium， and Korla pear gained a total of 33.61% of its total nitrogen from intercropping with alfalfa. This study offers a novel approach to enhancing nitrogen use efficiency， conserving land resources， and addressing fertilizer shortages in tree-forage tercropping system.

Effects of different forage reseeding patterns on plant community structure and soil characteristics in desert steppe

Ping-an BAO, Zhi-lin WEN, Yan WANG, Yan-hu CHEN, Bo JI, Zhan-jun WANG, Xu-dong WU, Qi JIANG

2025, 34(10):  62-73.  DOI: 10.11686/cyxb2024438

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We conducted an in-depth analysis of reverse succession in degraded desert grasslands to identify its driving factors. A field experiment was conducted in degraded desert grasslands of Ningxia， and the changes in plant community characteristics， soil physicochemical properties， and their correlations under different forage reseeding configurations （a grass mixture P₁， a legume mixture P₂， and a grass-legume mixture P₃） were investigated. The effects of reseeding configurations on plant community structure and soil nutrient contents were also evaluated. The results show that forage reseeding of the degraded pasture increased the proportion of legumes in plant communities， and resulted in increased vegetation cover and increased values of Simpson’s， Shannon-Wiener’s， and Margalef’s indexes. Reseeding with the grass-legume mixture increased the aboveground biomass of the plant community. Reseeding with the grass mixture significantly reduced soil bulk density， and reseeding with either the grass mixture or the grass-legume mixture decreased soil electrical conductivity. All the reseeding treatments resulted in increased total phosphorus and potassium contents in the 0-10 cm and 10-20 cm soil layers， increased organic carbon and total nitrogen contents in the 0-10 cm soil layer， and increased pH in the 10-20 cm layer； and decreases in soil pH in the 0-10 cm soil layer and organic carbon content in the 10-20 cm soil layer. The organic carbon and total nitrogen contents in the 0-10 cm soil layer showed positive correlations with plant community diversity indexes （Simpson’s， Shannon-Wiener’s， and Margalef’s indices）. Key factors driving vegetation restoration included organic carbon and total phosphorus contents in surface soil （0-10 cm）， along with pH and total phosphorus content in the subsurface soil layer （10-20 cm）. The grass-legume mixed configuration demonstrated optimal performance in enhancing grassland productivity， improving plant communities， and optimizing the soil nutrient profile. Therefore， reseeding with a grass-legume mixture is the preferred approach for restoration of degraded desert grasslands.

Planting and incorporation of smooth vetch together with reduced nitrogen fertilizer application enhances tobacco yield and soil quality

Wen-jun ZHAO, Ting LIANG, Jian-song WANG, Kui LIU, Yu FENG, Zheng-xu WANG, Zi-he XU, Yun-cong ZHU, Meng-meng SUN, Xiang-wei LI, Li-bo FU, Mei YIN, Guo-peng ZHOU, Hua CHEN, Wei-dong CAO

2025, 34(10):  74-84.  DOI: 10.11686/cyxb2024453

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The overall aim of our research was to provide a theoretical basis and technical approach to reduce fertilizer use and improve fertilizer use efficiency during tobacco production in Yunnan. To this end， we conducted an experiment in which different amounts of smooth vetch （Vicia villosa） were incorporated together with reduced application rates of nitrogen fertilizer to tobacco （Nicotiana tabacum）， and the yield and soil quality were evaluated. The field experiment commenced in 2017， and the impacts of planting and incorporating varying amounts of smooth vetch together with 30% and 15% reductions in the nitrogen fertilizer application rate on tobacco yield and soil quality were evaluated in 2022. Compared with the conventional fertilization practice of fallow-tobacco （control-F₁₀₀）， incorporating 22500 to 30000 kg·ha^-1 green manure with a 15% and 30% reduction in the nitrogen fertilizer application rate significantly increased tobacco yield by 4.85%-9.94% （P<0.05）. The incorporation of smooth vetch with reduced nitrogen fertilizer application also improved soil nutrient properties. Specifically， the treatment with 30000 kg·ha^-1 green manure incorporation and a 30% reduction in the nitrogen fertilizer rate resulted in the highest soil organic matter and soluble organic nitrogen contents， which were increased by 28.2% and 242.6%， respectively， compared with those in the control （P<0.05）. Incorporation of smooth vetch at rates of 22500 and 30000 kg·ha^-1， in conjunction with a 30% reduction in the nitrogen application rate， increased the soil quality index by 49.1% and 72.9%， respectively， compared with that in the control （P<0.05）. Under different nitrogen application levels， the activities of hydrolytic enzymes associated with soil carbon， nitrogen， and phosphorus increased with increasing amounts of incorporated green manure. Partial least squares path modeling and random forest analyses indicated that tobacco yield was primarily influenced by the soil organic matter content and enzyme activity. Therefore， planting and incorporating green manure with the linked reduction of nitrogen fertilizer enhanced both tobacco yield and soil quality， with outstanding effects observed at 22500 to 30000 kg·ha^-1 of smooth vetch incorporation with a 30% reduction in nitrogen fertilizer.

Effects of pine needle essential oil on the quality and aerobic stability of Broussonetia papyrifera silage

Qing-yuan DENG, Dong-qing FU, Rong-zheng HUANG, Fan-fan ZHANG, Guo-jun SUN

2025, 34(10):  85-94.  DOI: 10.11686/cyxb2024460

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This study aimed to elucidate the effects of pine needle （Pinus） essential oil on the quality and aerobic stability of Broussonetia papyrifera silage， and thereby provide technical data to facilitate its processing and utilization. B. papyrifera was fermented spontaneously with no addition of pine needle oil （CK）， or with addition of 160 mg·kg^-1 （Ⅰ）， 320 mg·kg^-1 （Ⅱ） or 480 mg·kg^-1 （Ⅲ） pine needle oil in a single-factor experiment. Various parameters， including the nutritional composition， fermentation quality and microbial count of the B. papyrifera silage were evaluated throughout the ensiling process （3， 7， 15， 30 and 60 days）. Additionally， aerobic stability was assessed at 60 days. It was found that on day 3 of ensiling， the water-soluble carbohydrate （WSC） content， lactic acid （LA） concentration， and acetic acid （AA） concentrations in groups Ⅰ， Ⅱ， and Ⅲ were significantly higher than those of CK （P<0.05）. Conversely， pH level， ammonia nitrogen∶total nitrogen （NH₃-N∶TN）， propionic acid （PA） concentration， and mold count were significantly lower in the treated silages than in CK （P<0.05）. Lactic acid bacteria counts and yeast counts in groups Ⅱ and Ⅲ differed significantly from those of CK （P<0.05）. On day 7 of ensiling， lactate concentration， acetic acid concentration， and lactic acid bacteria count in groups Ⅰ， Ⅱ， and Ⅲ were significantly higher than those in the CK group （P<0.05）. In contrast， pH level， NH₃-N∶TN， PA concentration， yeast counts and mold counts on day 7 were significantly lower in these groups than in CK （P<0.05）. Additionally， the dry matter （DM） content and WSC content in groups Ⅱ and Ⅲ were also significantly higher than those observed in CK （P<0.05）. On day 15， the crude protein （CP） content， WSC content， lactate concentration， acetic acid concentration and lactic acid bacteria count in groups Ⅰ， Ⅱ， and Ⅲ were significantly higher than in CK （P<0.05）. Conversely， pH level， NH₃-N∶TN， PA concentration， yeast count and mold count in groups Ⅰ， Ⅱ， and Ⅲ were significantly lower than in CK （P<0.05）. On day 30， the WSC contents， lactate concentrations， acetic acid concentrations， and lactic acid bacteria counts in groups I， Ⅱ， and Ⅲ were significantly elevated compared to CK （P<0.05）. Conversely， there was a significant reduction in the pH level， NH₃-N∶TN， PA concentration， and yeast counts compared to the CK （P<0.05）. On day 60， the CP content， WSC content， lactic acid concentration， acetic acid concentration， and lactic acid bacteria counts in groups I， Ⅱ， and Ⅲ were significantly higher than in CK （P<0.05）. As earlier， pH level， NH₃-N∶TN， PA concentration， and yeast counts were significantly decreased compared with the CK （P<0.05）. The aerobic stability of groups Ⅱ and Ⅲ was notably superior to that of both group I and CK （P<0.05）. In summary， pine needle essential oil effectively reduced the loss of CP and WSC during silage production. It increased lactate concentration， acetic acid concentration， and lactic acid bacteria count， while lowering pH level， and inhibiting mold proliferation. Furthermore， it significantly improved the aerobic stability of B. papyrifera silage. The addition of 480 mg·kg^-1 produced the most pronounced silage fermentation improvement effect.

Effects of exogenous ABA on growth and physiological characteristics of sainfoin seedlings under NaCl stress

Ge TIAN, Li-li NAN, Li-qun WANG, Xiang-xiang MA, Rong HE, Jia-yu GUO

2025, 34(10):  95-106.  DOI: 10.11686/cyxb2025022

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Soil salinization is a significant factor that limits plant nutrient uptake and affects plant growth and development. Sainfoin （Onobrychis viciifolia） is a native fabaceae species in saline-alkali regions of China， and exogenous abscisic acid （ABA） plays a positive regulatory role in enhancing plant stress resistance. This study investigates the effects of exogenous ABA on the growth and physiological characteristics of sainfoin seedlings subjected to salt stress. Using a nutrient solution sand culture method， various concentrations of ABA were sprayed on the leaves of both salt-tolerant （‘GH’， ‘GN’， ‘1994’） and salt-sensitive （‘10295’， ‘2323-2’， ‘2668’） sainfoin seedlings under a 0.8% NaCl treatment. Specifically， one control group （CK） and four treatment groups were established： 0.8% NaCl （T₁）， 0.8% NaCl+0.1 mmol·L^-1 ABA （T₂）， 0.8% NaCl+0.2 mmol·L^-1 ABA （T₃） and 0.8% NaCl+0.3 mmol·L^-1 ABA （T₄）. The optimal concentration of ABA for alleviating salt stress was identified. The results indicate that ABA has a mitigating effect on salt stress. That plant height， leaf area， aboveground biomass， underground biomass， relative leaf water contents， root activity， K⁺ contents， gibberellin contents in leaves and roots， zeatin contents in leaves and roots and auxin （IAA） contents in leaves decreased with increasing salt stress. The changes in the salt-tolerant material ‘GH’ were quite significant. The plant height increased by 20.48%， while the leaf area increased by 4.63%； Additionally， the aboveground biomass saw an increase of 26.75%， and the underground biomass increased significantly by 56.66%； The relative leaf water content rose by 28.19%， and root activity experienced a substantial increase of 65.41%. Furthermore， K⁺ content increased by 24.36%， leaf gibberellin content rose by 47.55%， and root gibberellin content increased by 36.05%； The leaf zeatin content increased by 24.39%， while leaf zeatin saw a notable rise of 80.53%， and leaf IAA content increased by 19.20%， respectively， in 0.8% NaCl， compared to control. Additionally， malonaldehyde contents， superoxide dismutase activity， peroxidase activity， catalase activity， Na⁺ contents， Ca²⁺ contents， ABA contents in leaves and roots and IAA contents in roots were reduced in response to salt. The changes in the salt-sensitive material ‘10295’ were relatively insignificant， with malonaldehyde contents， superoxide dismutase activity， peroxidase activity， catalase activity， Na⁺ contents， Ca²⁺ contents， ABA contents in leaves， ABA contents in roots and IAA contents in roots increasing by 14.91%， 18.64%， 26.15%， 10.08%， 24.47%， 30.24%， 8.72%， 27.64%， and 30.87%， respectively， in 0.8% NaCl stress. These results indicate that exogenous ABA can mitigate oxidative damage to cells caused by salt stress， maintaining the integrity of the cell membrane system and cell osmotic pressure. In this study， 0.2 mmol·L^-1 ABA demonstrated the most effective results， with a more pronounced response observed in salt-tolerant materials compared to salt-sensitive materials.

Effects of interactions among fertilizer， water retention agent， and seeding rate on the yield production performance of oat （Avena sativa）

Wen-yan MA, Jie-dong LI, Zhen-lei ZHOU, Dong CAO, Bao-long LIU, Huai-gang ZHANG, Dong-xia WANG

2025, 34(10):  107-119.  DOI: 10.11686/cyxb2024353

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The aim of this study was to determine the effects of interactions among fertilizer， polyacrylamide water-retention agent （PAC）， and seeding rate on the yield， yield components and nutritive traits of oat （Avena sativa） in the Hulunbuir region. A field experiment was conducted with an orthogonal design using the oat cultivar ‘Qinghai 444’. The experiment was an incomplete 3×3×3 factorial design with three fertilizer treatments （A₁， A₂， A₃）， three levels of PAC （B₁， B₂， B₃）， and three seeding rates （C₁， C₂， C₃） with nine of the 27 available treatment combinations implemented. The three fertilizer treatments were： A₁=N， 44.00 kg·ha?¹ and P， 83.00 kg·ha?¹， A₂=N， 44.00 kg·ha?¹ and P， 41.00 kg·ha?¹ and A₃=N， 89.00 kg·ha?¹， P： 41.00 kg·ha?¹. The three PAC treatments were 60， 75 and 90 kg·ha?¹ PAC for B₁， B₂ and B₃， respectively， and the three seeding rates were calculated to deliver 2.5， 3.5 and 4.5 live seeds·ha?¹ for C₁， C₂ and C₃， respectively. Agronomic traits， individual plant biomass， forage yield， and nutritional quality indicators were analyzed for each combination of treatments. The results revealed that the total plant biomass， stem biomass， crude protein， calcium， and phosphorus content were significantly higher with the A₃ level of fertilizer than with the A₁ and A₂ levels of fertilizer. As the concentration of PAC increased， the seedling number， spike number， tiller survival rate， sub-spike internode length， plant height， center of gravity height， lodging rate， and ash content increased， and total plant biomass， root biomass， and leaf biomass decreased. As the seeding rate increased， the stem number， spike number， center of gravity height， root biomass， and neutral detergent fiber content increased， whereas the grain number per plant， sub-spike internode length， spike length， plant height， spike position height， total plant biomass， spike biomass， and acid detergent fiber content decreased. Among the nine treatment combinations， the A₃B₁C₃ treatment resulted in the highest stem number， second and third internode length， stem diameter， stem wall thickness， crude protein， ether extract， calcium， and phosphorus content， along with the lowest acid detergent fiber content. The A₃B₂C₁ treatment resulted in the highest spike position， total plant biomass， stem biomass， and spike biomass. The results of a range analysis indicated that fertilizer application had the greatest impact on oat yield and quality at the flowering stage， followed by the water-retention agent， while seeding rate had the smallest effect. A multivariate evaluation using the membership function method identified the optimal combination for oat yield and quality performance as A₃B₁C₃； i.e.， a fertilizer ratio of N： 89.00 kg·ha?¹， P： 41.00 kg·ha?¹， K： 41.00 kg·ha?¹， application of PAC at 60.00 kg·ha?¹， and a seeding rate of 157.05 kg·ha?¹. These findings provide a theoretical basis and technical guidance for sustainable and high-efficiency oat cultivation in the Hulunbuir region.

Effects of nitrogen fertilizer reduction combined with humic acid on soil fungal communities in drip irrigated maize fields in northern Xinjiang

Xiao-qian LU, Jin-lu CHEN, Wei-jun YANG, Qing-yun GUO, Dan-li WANG, Hong-mei ZHAO

2025, 34(10):  120-131.  DOI: 10.11686/cyxb2024472

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The objective of this study was to evaluate the effect of humic acid addition on soil fungal communities in drip irrigated maize （Zea mays） fields in northern Xinjiang under reduced nitrogen fertilizer supply， and to explore the optimal application of nitrogen fertilizer in farmland. In this study， a randomized block experiment was adopted， and six treatments were set up： No nitrogen fertilizer （CK， 0 kg·ha^-1）， conventional nitrogen fertilizer （T₁， 300 kg·ha^-1）， humic acid application alone （T₂， 90 kg·ha^-1）， conventional nitrogen fertilizer combined with humic acid （T₃）， nitrogen fertilizer reduction of 15% combined with humic acid （T₄）， and nitrogen fertilizer reduction of 30% with humic acid （T₅）. Illumina MiSeq high-throughput sequencing technology was used to analyze the changes in fungal diversity and community structure under the six different treatments， and to explore the effects of nitrogen fertilizer reduction combined with humic acid application on soil fungal community composition. Supporting data on soil nutrient levels and maize yield were also collected. It was found that the soil electrical conductivity （EC） of T₄ and T₅ was significantly reduced by 18.35% and 24.85%， respectively， compared with T₁， and the soil alkaline hydrolyzable nitrogen content was significantly increased by 32.00% and 18.40%， respectively under the same treatments. The maize yield was highest （18038.75 kg·ha^-1） in the T₄ treatment； however， the treatments had no significant effects on the soil fungal community Alpha and Beta diversity. With respect to soil fungi， the relative abundance of Ascomycota under T₄ and T₅ was significantly increased by 21.85% and 24.59%， respectively， compared with T₂， but the relative abundance of Mortierellomycota and Chytridiomycota was significantly reduced. Pearson correlation analysis showed that soil conductivity was significantly positively correlated with Ascobolus and negatively correlated with Botryotrichum， while soil organic matter was negatively correlated with Fusarium presence， and soil alkaline hydrolyzable nitrogen was significantly positively correlated with Cladosporum presence. Further redundancy analysis showed that soil available phosphorus was the main environmental factor affecting the community structure of fungi in this drip irrigated maize field， while soil EC value and alkali hydrolyzable nitrogen were the main environmental factors affecting the horizontal community structure of soil fungi. In conclusion， nitrogen fertilizer reduction combined with humic acid changed the soil fungal community structure， promoted the growth of beneficial bacteria， and inhibited the reproduction of pathogenic fungi by causing changes in soil nutrient content. This information is of great significance for improving the soil microbial environment， maintaining the balance of the soil microbial community and increasing crop yield.

Studies of the difference in symbiotic interaction between Sinorhizobium meliloti strain QL2 and different alfalfa varieties

Yi-lin HAN, Wen-juan KANG, Shang-li SHI, Yuan-yuan DU, Fu-qiang HE, Yan WANG, Wen-lu HOU, Xi-lin XIE

2025, 34(10):  132-150.  DOI: 10.11686/cyxb2025030

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Inocculum of Sinorhizobium meliloti strain QL2 was applied to plants of eight alfalfa varieties （three imported varieties： WL168HQ， WL298HQ， and WL319HQ； three domestically bred varieties： Gannong No. 3， Gannong No. 5， and Gannong No. 9； and two local varieties： Qingshui and Longzhong） to clarify the differences in nitrogen fixation effects， with a view to improving the nodule formation and nitrogen fixation capacity of alfalfa and the efficiency of natural nitrogen utilization. Indicators such as nodule formation， nitrogen fixation， forage biomass， and nutritional quality were evaluated to study the differences in symbiosis， nitrogen fixation， and growth-promoting effects after inoculation with S. meliloti QL2. Results demonstrated that in domestically bred varieties， inoculating with S. meliloti QL2 significantly increased the weight of individual root nodules， as well as the root nodule diameter and the number of infected cells in the root nodules of by over 70% compared to both local and imported varieties. Regarding nitrogen fixation efficiency， the imported varieties （52.62% to 63.49%） and domestically bred varieties （53.30% to 62.41%） exhibited significantly higher fixation rates compared to local varieties （43.05% to 46.72%）. Hierarchical segmentation analysis showed that nodulation factors （the number of infected cells in the root nodules， the number of effective root nodules per plant， the single effective root nodule weight and root nodule diameter） and nitrogen fixation factors （nitrogenase activity， nitrogen fixation percentage and nitrogen fixation amount） explained 63% of the variation in the above-ground dry weight. Of the explained variation， 89.56% was contributed by nitrogen fixation factors， and only 13.55% by nodulation factors. A correlation analysis revealed that after inoculation with S. meliloti QL2， the nitrogen fixation percentage of different categories of variety was significantly positively correlated with above-ground dry weight. That is， the growth rate （as reflected by above-ground dry weight） of imported varieties was the largest compared with CK， of which WL319HQ-QL2 had the highest growth rate of 48.08 %， followed by domestic bred varieties and local varieties. The nitrogen fixation percentage was also significantly positively correlated with the forage nutritional quality. Inoculation with rhizobia generally increased the crude protein yield of alfalfa with the highest increase being 51.08% （P<0.05）， while reducing the yields of neutral detergent fiber and acid detergent fiber by 40.55% and 53.96%， respectively. These changes in nutritional composition resulted in an elevation of the forage quality score by 1 to 2 grades. In summary， a high nitrogen fixation rate has a significant positive impact on both above-ground dry weight and forage nutritional quality， meaning that it plays the critical role in improving the yield and nutritional quality of alfalfa. This study lays a foundation for development of materials to optimize nitrogen fixation efficiency in legume-rhizobia symbioses and improve alfalfa forage quality.

Optimization of culture conditions for the sophoridine-producing new fungal species Achaetomium sophor strain HY17 isolated from seeds of Sophora alopecuroides

Zhi-yun TANG, Wen-kai WANG, Guan-lan LIU, Pei-wen GU

2025, 34(10):  151-163.  DOI: 10.11686/cyxb2024444

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The aim of this study was to screen for endophytic fungal strains of Achaetomium sophora infecting Sophora alopecuroides that produce sophoridine alkaloid， classify and identify them， and then identify in-vitro culture conditions that optimize the alkaloid production of the endophyte fungus. The overall goal was to improve alkaloid yield and obtain excellent strains for culture. Fifty strains of A. sophora isolated from healthy S. alopecuroides seeds were screened using alkaloid precipitation， acid dye colorimetry， and high-performance liquid chromatography analyses. The strains were identified through morphological and molecular analyses. Subsequently， a series of methods including the single factor test， Plackett-Burman （PB） test， steepest climb test， and response surface test （Box-Behnken test） were used to investigate the effects of medium composition （medium type， carbon and nitrogen sources）， culture conditions （incubation time and pH）， and precursors and inducers on the alkali yield of selected fungal strains. From the results of these analyses， the optimal alkali-producing medium， culture conditions， and precursors for sophoridine alkaloid production were determined. The fungal strain HY17 producing sophoridine at high levels was identified as a new species， Achaetomium sophora. The optimal culture conditions for A. sophora HY17 were determined to be SDY liquid medium with an initial pH of 6 and a culture period of 8 days， with maize flour as the carbon source and casein as the nitrogen source. Tyrosine， L-lysine， L-piperidinic acid， and phenylalanine had significant effects on sophoridine production by strain HY17 during culture. The addition of L-lysine （1.044 g·L^-1）， L-piperidinic acid （0.081 g·L^-1）， and phenylalanine （1.995 g·L^-1） resulted in a maximum alkaloid yield of 1.369 mg·g^-1 mycelium under these optimized conditions， representing a remarkable increase of 61.95% compared with that obtained under the original culture conditions. A. sophora HY17 produced sophoridine at a high and stable rate. Optimization of the culture conditions resulted in a notable increase in alkaloid production. The results of this study offer a novel approach for sophoridine production through in-vitro culture of the fungal endophyte， A. sophora strain HY17 isolated from S. alopecuroides.

Functional identification of the role of soybean gene GmPP2C28 in the nitrogen-fixation process of Lotus japonicus

Dan-xia KE, Shi-bo HOU, Zhao-yuan ZHOU, Yun-hao MA, Zhi-jie CHEN, Xiao-li SONG, Jia-nuo LIN

2025, 34(10):  164-173.  DOI: 10.11686/cyxb2024427

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Protein phosphatase 2C （PP2C） is a crucial serine/threonine protein phosphatase in plants. These enzymes are encoded by a large gene family， and they play key roles in plant development and various stress responses. However， there is relatively little research on the role of PP2Cs in the formation of nodules in the symbiotic relationship between leguminous plants and rhizobia. Previously， a soybean （Glycine max） PP2C family gene GmPP2C28 was cloned and its expression was confirmed to be induced by rhizobia. In this study， we constructed a plant overexpression vector p1302G-GmPP2C28 and obtained transgenic Lotus japonicus plants using Agrobacterium tumefaciens LBA1334-mediated transformation of hairy roots. The transgenic GmPP2C28-expressing plants formed significantly more nodules than did those transformed with the empty vector， and the transcript level of the nodule indicator gene also tended to be higher in the transgenic plants than in the empty vector control plants. Methylamine blue staining of nodule slices revealed that overexpression of GmPP2C28 significantly increased the number of bacteroids in the infected area of the nodules. Analyses of nodule nitrogenase activity revealed that overexpression of GmPP2C28 significantly increased nitrogenase activity in mature and senescent nodules. Together， these results show that heterologous expression of GmPP2C28 in L. japonicus significantly increased the numbers of nodules and filamentous bacteria in the root nodules. In addition， overexpression of GmPP2C28 significantly increased nitrogenase activity in mature and decaying nodules， thereby greatly delaying nodule aging. The findings of this study identify a new candidate gene for creating excellent varieties of L. japonicus and other legume species with strong biological nitrogen fixation.

Ion balance mechanism and transcriptome analysis of Elytrigia elongata in response to NaHCO₃ stress

Ran ZHANG, Chen-zhuo LIU, Feng YUAN, Ya-ling LIU, Di DONG, Si-ning WANG, Bo-kun ZOU, Xiao-xia LI

2025, 34(10):  174-186.  DOI: 10.11686/cyxb2024456

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Elytrigia elongata， due to its strong salt-alkali tolerance， is widely used to establish salt alkali pastures. This research explored the ion balance mechanism of E. elongata in response to salt-alkali stress， using the cultivar ‘Orbit’ as the experimental material， and 150 mmol·L^-1 NaHCO₃ to simulate alkali stress. The growth indexes and mineral contents （Na⁺， K⁺， Ca²⁺， Mg²⁺， Cl^-， SO₄^2- and NO₃^-） of seedlings were determined， and high-throughput Illumina Hiseq sequencing technology was used to perform transcriptomic analysis on leaves and roots after exposure to contrasting normal （CK） growth conditions and NaHCO₃ treatment for 24 hours. It was found that the root biomass and root-shoot ratio of E. elongata increased significantly under NaHCO₃ stress. RNA-Seq results showed that there were 1833 differentially expressed genes （DEGs） in the leaves and 1536 DEGs in the roots under NaHCO₃ treatment， and 140 genes were differentially expressed in both leaves and roots. GO and KEGG enrichment analysis revealed that DEGs in leaves and roots were significantly enriched in metabolic pathways related to antioxidant synthesis， ion binding， and phenylalanine， and phenylpropanoid biosynthesis. DEGs related to ion binding pathways included BAK1， CIPK10， STRK1， WAK8， and multiple laccase genes （laccase-11 and laccase-3）， which may be involved in the response process of E. elongata to NaHCO₃ stress. In addition， physiological test results further demonstrate that the ion transport and distribution in the leaves and roots of E. elongata are affected. This was evidenced by a large accumulation of Na⁺ and a decrease in ability to absorb K⁺， resulting in a decrease in the ratio of Na⁺∶K⁺. The roots maintained tissue ion balance by improving the absorption and distribution capacity of other cations such as Ca²⁺， Mg²⁺ and Fe³⁺， so as to better adapt to the saline-alkali environment. The results of this study identify specific genes involved in salt-tolerance， and will be useful to inform molecular breeding of forage and other crops. These results also provide a theoretical underpinning for the promotion of E. elongata and the improvement and utilization of saline-alkali land.

Identification of the chitinase gene family and functional analysis of TrChit3 from white clover

Xiao-fang QIN, Zhi-rui HE, Tong JIA, Yu-jiao YANG, Wei FU, Hang LI, Yan PENG

2025, 34(10):  187-201.  DOI: 10.11686/cyxb2024424

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Chitinases are glycoside hydrolases that play an important role in plant growth and development. Relatively few studies have focused on the responses of chitinases to various biotic and abiotic stresses in white clover （Trifolium repens）. Therefore， in this study， we searched the whole genome of white clover to identify and characterize its chitinase gene family. A total of 44 chitinase genes were identified in the whole genome of white clover， and they were unevenly distributed on 16 chromosomes. Phylogenetic analysis of these chitinase genes classified them into two mega-groups （GH18 and GH19）， with five separate clusters （I-V）. Twenty-nine chitinase genes belonged to the GH18 subfamily and 15 belonged to the GH19 subfamily. The genes within the same subfamily genes encoded proteins with similar structures and functions. Promoter analysis showed that the promoter regions of these 44 chitinase genes contained a variety of cis-acting elements related to growth and development， hormones， and stress. Furthermore， quantitative reverse-transcription polymerase chain reaction analyses showed that salt stress and drought stress resulted in significant increases in the transcript levels of chitinase genes， which further verified their roles in the plant stress response. TrChit3 was selected for a detailed analysis of its role in drought resistance. Overexpression of TrChit3 in Arabidopsis thaliana enhanced its resistance to drought stress， as measured by changes in physiological indexes such as relative conductivity and malondialdehyde content. These results lay a foundation for further studies on the roles of chitinases in plants.

Effect of different levels of purple maize anthocyanin extract on in vitro gas production and rumen fermentation in beef cattle

Qi LU, Ji-xiao QIN, Yi-ming BAN, Cheng-cheng GAO, Rong YANG, Pei-yao LI, Yi-qing XU, Shuang-long XIE, Rui CHEN, Di ZHOU, Xing-zhou TIAN

2025, 34(10):  202-212.  DOI: 10.11686/cyxb2024485

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The object of this study was to investigate the effect of purple maize anthocyanin extract on in vitro gas production and rumen fermentation in beef cattle. A completely randomized design was used. Levels of 0， 0.2%， 0.4%， 0.6%， and 0.8% purple maize anthocyanin extract were added to the substrate， and total gas production， gas production kinetic parameters， volatile fatty acid， methane production， cellulase activity， and nutrient degradation rate were measured. It was found that： 1） Total gas production after fermentation for 72 h tended to be reduced where anthocyanin was added， and gas production at 72 h for the 0.6% anthocyanin group was significantly lower （P<0.05） than that of the control group. 2） By contrast， gas production from the immediately soluble fraction tended to be higher with anthocyanin present and was significantly higher （P<0.05） in the 0.4% anthocyanin group than in the control group. The gas production rate constant decreased with increase in the purple maize anthocyanin extract supplemental level， and the control group was significantly higher （P<0.05） than the other four anthocyanin groups， and the curve defining the trend across anthocyanin levels had both linear and quadratic significant （P<0.05） terms. 3） The contents of acetic and butyric acid at 24 h decreased with increase in added anthocyanin， with significant （P<0.05） linear and quadratic terms for the fitted curve. Accordingly， the contents of acetic and butyric acids in the control group were significantly higher （P<0.05） than those of the 0.4%， 0.6%， and 0.8% anthocyanin groups. In contrast， the content of propionic acid increased with increase in supplemental purple maize anthocyanin extract， with both linear and quadratic terms of the fitted curve being significant （P<0.05）， and accordingly， the content of propionic acid in the control group was significantly lower （P<0.05） than other anthocyanin groups. The acetic：propionic acid ratio and methane production both decreased with increase in the level of anthocyanin extract and in both cases the fitted curve had significant （P<0.05） linear and quadratic terms. Accordingly， acetic：propionic acid ratio and methane production in the anthocyanin groups were significantly lower （P<0.05） than that of the control group. 4） At 12 h fermentation time， carboxymethyl cellulose and cellobiase were increased with increase in the supplemental anthocyanin extract level and both linear and quadratic terms were significant （P<0.05）. Accordingly carboxymethyl cellulose and cellobiase levels in the 0.8% anthocyanin group were significantly higher （P<0.05） than in the other four groups. Meanwhile， the values of xylanase in the 0.2%， 0.4%， and 0.6% anthocyanin groups were significantly higher （P<0.05） than those in the control group at 24 h fermentation time. 5） Both crude protein degradation rate and acid detergent fiber degradation rate increased with increase in supplemental anthocyanin extract level， and linear and quadratic terms of the fitted curves were significant （P<0.05） increasing. Accordingly， the crude protein degradation rate in the anthocyanin groups was significantly higher （P<0.05） than that of the control group. Taken together， purple maize anthocyanin extract had the ability to improve rumen fermentation mode， increase propionic acid， cellulase activity， and nutrient degradation rate， and decrease methane production of beef cattle. Under the conditions of this experiment， the optimal purple maize anthocyanin extract level was 0.4%.

Effects of substituting mulberry silage for soybean meal on blood indexes， rumen fermentation， and rumen bacteria of goats

Peng-fei CAO, Shui-ping WANG, Qiao-shen HUANG, Shi-long ZHOU, Zhuan LUO, Ying REN, Yong LIU, Tie-jun LI, Shao-xun TANG

2025, 34(10):  213-228.  DOI: 10.11686/cyxb2024426

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We investigated the effects on the blood biochemical indexes， antioxidant indexes， rumen fermentation indexes， and rumen bacteria of two different ratios of mulberry silage as a replacement for soybean meal in the diet of Xiangdong black goats. Forty-five healthy Xiangdong black goat rams aged 5-6 months with similar body weight （18.2±1.6 kg） were randomly divided into three groups： a control group （group CK）， a 50% replacement group （group S₁） and a 100% replacement group （group S₂）. The pre-test period was 7 days， and the formal experimental period was 55 days. The remaining feed from the previous day was weighed every morning before feeding and the goats were weighed every 2 weeks. On the morning of the 54^th day of the formal test period， blood samples were collected from the jugular vein of each goat. Rumen fluid was collected orally 3 hours after morning feeding on the 54^th day and before morning feeding on the 55^th day of the formal experimental period for the determination of blood indexes， rumen fermentation indexes， and rumen microbial composition. The results showed that the final body weight， average daily gain， and dry matter intake were significantly higher in the S₁ and S₂ groups than in the CK group （P<0.05）. The serum low density lipoprotein cholesterol concentration was significantly lower in group S₂ than in the CK group and group S₁ （P<0.05）， and serum albumin and cholinesterase concentrations were significantly higher in group S₂ than in the CK group （P<0.05）. The total antioxidant capacity was significantly higher in groups S₁ and S₂ than in the CK group （P<0.05）， but the malondialdehyde content was significantly higher in group S₂ than in the CK group （P<0.05）. As the amount of mulberry protein in the diet increased， the molar ratio of acetic acid to isovaleric acid decreased significantly （P<0.05）， and the molar ratio of propionic acid to valeric acid increased. The replacement of soybean meal in the diet with mulberry silage did not affect the alpha diversity of rumen microorganisms （P>0.05）. The dominant taxa in the three groups were Bacteroidota and Bacillota. With increasing amounts of mulberry silage in the diet， the abundance of Bacillotaincreased significantly （P<0.05）， and the abundance of Bacteroidota decreased significantly （P<0.05）. The dominant bacteriain the threegroups were Xylanibacter， Butyrivibrio， Segatella，Succiniclasticum， Fibrobacter， and Olivibacter. The abundance of Butyrivibrio was significantly higher in group S₂ than in the CK group and group S₁ （P<0.05）， and that of Olivibacter was significantly lower in group S₂ than in the CK group and group S₁ （P<0.05）. These results show that replacing soybean meal with mulberry silage in the diet of goats improved their metabolism and increased their antioxidant capacity without affecting the alpha diversity of their rumen microorganisms. Therefore， this resource has promising applications in ruminant breeding.