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01 July 2024, Volume 57 Issue 13

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Scientia Agricultura Sinica. 2024, 57(13):  0. 

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Analysis of Heterosis and Combining Ability of Lodging Resistance Traits of Spring Wheat Varieties (Lines) in the Ningxia Yellow River Irrigation Area

SUN WeiHao, LIU Ting, SANG YiNan, YANG ZhengWei, ZHANG GaiSheng, SONG YuLong, ZHANG ShuangXi

Scientia Agricultura Sinica. 2024, 57(13):  2497-2508.  doi:10.3864/j.issn.0578-1752.2024.13.001

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【Objective】 The essence of conventional wheat breeding involves the continuous multi-generation selection and maintenance of hybrid F₁ trait heterosis. Exploring the heterosis and combining ability of lodging-related traits among spring wheat varieties (lines) in the Yellow River irrigation area of Ningxia, and clarifying significantly agronomic indexes related to anti-lodging traits in wheat that can provide a certain theoretical basis for utilizing heterosis in lodging traits of spring wheat, and for the screening and maintenance of its descendants in Ningxia.【Method】 This study utilized 14 spring wheat varieties (lines) from Ningxia Yellow River Irrigation Area as parent materials. According to NCⅡ incomplete diallel crossbreeding design, forty-five combinations were produced. Meanwhile, heterosis, combining ability, and correlation analyses were performed on 13 traits related to lodging resistance, such as plant height and culm type index, in their parents and their F₁ generation.【Result】 Significant differences in lodging resistance trait were observed among different spring wheat varieties (lines). Meanwhile, there is a certain level of heterosis in lodging resistance traits was evident in the F₁ generation. The main stem thrust resistance of NZ42, M6445, and M8887 showed high general combining ability (GCA) values of 11.68, 8.00, and 10.67 respectively. Significant combinations for main stem thrust resistance included M6445×M8887, M6445×MJ48, NZ42×N2038, NZ42×M7723, NZ42×NZ39, and H3015×Ningchun50. Correlation analysis revealed that plant height was significantly positively correlated with the first internode length and second internode weight, and highly significantly positively correlated with the second and fourth internode lengths. Besides, the main stem thrust resistance was highly significantly positively correlated with fresh weight of the main stem, stem diameter, bending moment and culm type index, but significantly negatively correlated with the fourth internode length. However, it was not significantly correlated with the second internode weight and fullness.【Conclusion】 The excellent parent varieties identified were NZ42, M6445, and M8887. Meanwhile, the excellent combinations of NZ42 × NZ39, M6445 × MJ48, and M6445 × M8887 were also observed in this study. In addition, Heterosis was present among most lodging resistance traits in different combinations, but the heterobeltiosis was less pronounced. The resistance to lodging was influenced by both additive and non-additive effects of parental genes, with a greater influence from the maternal genetic background. Furthermore, the plant height and main stem thrust resistance are significantly correlated with the fourth internode length, bending moment, and culm type index, serving as important references for selection of lodging-resistant generations.

Identification of Resistant Germplasms and Mining of Candidate Genes Associated with Resistance to Stalk Rot Caused by Synergistic Infection with Fusarium spp. in Maize

GUI CuiLin, MA Liang, WANG YinYing, XIE FuGui, ZHAO CaiHong, WANG WenMiao, LI Xin, WANG Qing, GAO XiQuan

Scientia Agricultura Sinica. 2024, 57(13):  2509-2524.  doi:10.3864/j.issn.0578-1752.2024.13.002

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【Objective】 Maize stalk rot is one of the common and most devastating diseases in major maize production areas in China. Under natural conditions, maize stalk rot is mostly caused by the synergistic infection of various pathogens. The purpose of this study was to screen elite maize germplasms resistant to the synergistic infection with F. graminearum and F. verticillioides. Meanwhile, QTN associated with resistance to the infection by synergistic infection of two Fusarium species and the candidate resistance genes were identified, which will provide gene resources and theoretical reference for molecular breeding of maize variety resistant to synergistic infection of different pathogens.【Method】 Using a maize natural population as experimental materials, the maize stalk rot phenotypes were investigated upon simultaneous infection with F. graminearum and F. verticillioides. Furthermore, GWAS analysis was conducted to identify significant resistance SNP and to predict candidate genes for stalk rot resistance.【Result】 Through the phenotypic analysis of stalk rot caused by synergistic infection under both field and laboratory conditions, it was found that inbred lines from different sources and subgroups showed significant phenotypic variation. More specifically, the field assay results showed that the inbred lines collected from China were more resistant and that from USA were more susceptible to synergistic infection. Moreover, the inbred lines of tropical and subtropical subgroup were more resistant, whereas the inbred lines of Mixed subgroup more susceptible. The seedling assay results under laboratory condition showed that the lines collected from USA were more resistant, whereas that from CIMMYT were more susceptible. The lines of SS subgroup were more resistant, and that of Mixed subgroup were more susceptible. By integrating field and laboratory phenotypic data, 29 and 16 lines with higher levels of resistance to synergistic infection were screened out respectively, and 6 resistant lines were identified under both conditions. Moreover, based on the field phenotype GWAS, 18 QTNs associated with the resistance were identified, and 93 candidate genes associated with stalk rot resistance to synergistic infection were mined. Among these candidate genes, four genes showed haplotype variation, whose expression levels were up-regulated in disease-resistant lines.【Conclusion】 Using the natural population of maize with great diversity in genetic background, 6 resistant lines to the synergistic infection with Fusarium spp. were identified under two conditions, which can be used as potential germplasm resources for maize stalk rot resistance in the future. Four candidate genes that might be involved in the resistance to synergistic infection were identified by GWAS, which will provide genetic resources for the breeding maize varieties with enhanced resistance to stalk rot caused by F. graminearum and F. verticillioides.

Jasmonic Acid Regulates the Changes of Major Metabolites in Potato Tuber Development in vitro

LIANG LiJuan, CHENG LiXiang, YUAN JianLong, SA Gang, ZHANG Feng

Scientia Agricultura Sinica. 2024, 57(13):  2525-2538.  doi:10.3864/j.issn.0578-1752.2024.13.003

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【Objective】 Jasmonic acid (JA) is one of key regulatory hormones during potato tuber development. Study on JA regulating tuber development mechanism would provide an important theoretical basis for tuber yield and quality formation.【Method】 In this study, potato stolons cultured in vitro were exogenously applied with JA (0, 0.5, 5, 50 μmol·L^-1) to analyze the phenotype, tissue microstructure, carbohydrate accumulation and proteome change of tubers.【Result】 With the increase of JA concentration, the tuber number formed per stolon, tuber diameter, fresh and dry weight, cell area of perimedullary zone, starch and soluble sugar content were gradually increased at 0.5 and 5 μmol·L^-1 JA (P<0.05), whereas the tuber diameter, fresh and dry weight, starch content were significantly decreased at 50 μmol·L^-1 JA (P<0.05). Lipoxygenase activity was gradually decreased with JA concentration increasing (P<0.05). Thirty-five differentially abundant proteins (P<0.05 and fold change≥2.5) closely related to tuber development regulating by JA were identified by 2-DE and MALDI-TOF/TOF-MS. They were mainly involved in bioenergy and metabolism (28.6%), cell defense and rescue (28.6%), protein biogenesis and storage (11.4%), signaling (8.6%), transcription (8.6%), unknown (8.6%) and miscellaneous (5.6%). The differential expression patterns of these proteins were clustered into three categories by hierarchical clustering analysis. Cluster 1 included 17 proteins that down-regulated at 0.5 μmol·L^-1 JA and up-regulated at 5 μmol·L^-1 JA, which mainly involved in bioenergy and metabolism, protein biogenesis and storage, signaling and transcription. Cluster 2 included 10 proteins that gradually up-regulated with JA concentration increasing, which were mainly involved in cell defense and rescue, bioenergy and metabolism, transcription. Cluster 3 included 8 proteins that down-regulated after JA treatments, which were mainly involved in bioenergy and metabolism, cell defense and rescue, protein biogenesis and storage. 【Conclusion】 Low JA concentration (0.5, 5 μmol·L^-1) might promote tuber morphogenesis by inducing cell enlargement in tuber perimedullary zone, intracellular sucrose and polysaccharide accumulation, and cell defense ability, whereas high JA concentration (50 μmol·L^-1) showed inhibitory effects.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

The Differences of Plant Nitrogen Transport and Grain Nitrogen Accumulation in Different Yield Levels of Wheat Field

ZHANG Zhen, SHI Yu, YU ZhenWen, ZHANG YongLi

Scientia Agricultura Sinica. 2024, 57(13):  2539-2548.  doi:10.3864/j.issn.0578-1752.2024.13.004

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【Objective】 Improving the nitrogen transport and accumulation of nutrient organs after wheat flowering, as well as improving the nitrogen accumulation characteristics of wheat grains, is beneficial for increasing the nitrogen accumulation and yield of wheat. This study aimed to clarify the physiological mechanisms underlying the differences in nitrogen utilization efficiency among wheat plants at different yield levels, so as to provide a theoretical basis for reducing yield differences and improving nitrogen utilization efficiency and to achieve high yield and efficient production of wheat.【Method】 The field experiments were conducted in Shiwang Village, Xiaomeng Town, Shandong Province for two consecutive years from 2020 to 2022. Using Tobacco Farmer 1212 as the planting material, three yield levels of wheat fields with a perennial winter wheat yield of around 10 500 kg·hm^-2 (S), 9 000 kg·hm^-2 (H), and 7 500 kg·hm^-2 (M) were selected to compare and analyze the differences in plant nitrogen accumulation and transport, grain nitrogen accumulation characteristics, and grain yield in wheat fields with different yield levels.【Result】 Compared with H and M wheat fields, S wheat field significantly increased the number of spikes per unit area and thousand grain weight, with grain yield 19.64%-27.91% and 51.68%-80.87% higher than H and M wheat fields, respectively, resulting in the highest nitrogen absorption efficiency, nitrogen partial productivity and nitrogen harvest index. The nitrogen accumulation in the nutrient organs of S wheat field during the flowering period increased by 14.22-42.11 kg·hm^-2 and 53.74-103.16 kg·hm^-2, respectively. The nitrogen accumulation in mature nutrient organs was S, H>M. Compared with H and M wheat fields, S wheat field significantly increased the nitrogen transport of pre flowering nutrient organs and nitrogen accumulation after flowering, and increased the contribution rate of nitrogen accumulation in post flowering nutrient organs to grains, thereby achieving the highest nitrogen accumulation in mature grains. The S wheat field significantly increased the content of free amino acids and soluble proteins in flag leaves after flowering, for promoting the inter source sink transport of nitrogen. According to the logistic equation fitting, the S wheat field significantly increased the maximum and average accumulation rates of grain nitrogen, prolonged the duration of grain nitrogen accumulation, and was the main reason for obtaining the highest grain nitrogen accumulation.【Conclusion】 Suitable soil environment in S wheat field could promote the transfer of stored nitrogen from vegetative organs to grain, and increase the rate of grain nitrogen accumulation and prolong the duration of grain nitrogen accumulation, which was the main reason for the highest grain yield and nitrogen use efficiency.

Effects of Partial Replacement of Chemical Fertilizers with Green Manure on Dry Matter Accumulation and Yield Formation of Maize

QIN WenLi, ZHI JianFei, XIE Nan, ZHANG LiFeng, LIU ZhongKuan, LIU ZhenYu, FENG Wei, PAN Xuan, DAI YunXia

Scientia Agricultura Sinica. 2024, 57(13):  2549-2567.  doi:10.3864/j.issn.0578-1752.2024.13.005

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【Objective】 Replacing some chemical nitrogen fertilizers with leguminous green manure is an important technical measure to achieve reduced fertilizer application. The effects of hairy vetch which was planted to replace winter wheat and nitrogen reduction on the accumulation and transportation of dry matter accumulation, yield formation, and post flowering leaf senescence characteristics of maize in North China were studied, so as to provide a scientific basis for leguminous winter green manure substitution for chemical nitrogen fertilizer and optimized management of nitrogen resources in maize. 【Method】 The two-factor split plot field experiment was carried out from 2020 to 2022 at Shenzhou Experimental Station of Dryland Farming Institute of Hebei Academy of Agriculture and Forestry Sciences. The two modes, including the winter fallow field (FF) and hairy vetch being planted in the winter fallow field and total returning (HV), were set as main treatment, and the five nitrogen application rates of maize were set as sub-treatment, including no nitrogen application (0 N), 67.5 kg·hm^-2 (25% N), 135.0 kg·hm^-2 (50% N), 202.5 kg·hm^-2 (75% N), and 270.0 kg·hm^-2 (100% N, the conventional nitrogen application level). Yield and yield components, ear agronomic traits, dry matter accumulation and transport, leaf senescence characteristics of maize, and changes in soil nutrients and enzyme activity were investigated, moreover, the nitrogen surplus in farmland were analyzed.【Result】 Returning hairy vetch to the field significantly increased maize yield and could compensate for the reduction in maize yield caused by chemical nitrogen reduction. From 2021 to 2022, the return of hairy vetch dramatically raised maize yield (8.15%-9.21%), which could compensate for the grain yield loss caused by 25%-50% reduction in nitrogen fertilization. Continuous return of hairy vetch significantly reduced the bare top length, dramatically increased the ear length, the ear diameter, and yield components, such as row number, kernels number per row, and hundred-grain weight. After returning hairy vetch to the field, the dry matter accumulation before and after flowering, the dry matter accumulation rate after flowering, and the contribution rate of dry matter after flowering to grains significantly increased by 10.21-12.32 g/plant, 39.94-72.37 g/plant, 4.67%-4.78%, and 3.31%-3.99%, respectively, which could compensate for the negative effect of reducing nitrogen fertilizer application by 25%-50% on the dry matter accumulation before flowering and the contribution rate of dry matter after flowering to grains of maize. The incorporation of hairy vetch prominently postponed the post-anthesis leaf senescence of maize. In 2021 and 2022, the green leaf area of maize treated with HV was 303.44-1 115.10 and 266.23-837.62 cm²/plant higher than that under FF, respectively. The relative green leaf area after flowering increased by 1.12% -13.84% and 0.56% -9.13%, respectively. V_max decreased by 0.30% and 0.05%, respectively. T_max was delayed by 6.01 days and 3.56 days, respectively. The organic matter content, total nitrogen content, alkaline nitrogen content, as well as the activities of urease, sucrase, amylase, and protease in the soil significantly increased after continuous returning of hairy vetch to the field. In 2021 and 2022, the nitrogen rates under HV could increase by 40.92 and 72.79 kg·hm^-2, respectively; the optimal chemical nitrogen applications under HV could be reduced by 40.96 and 48.90 kg·hm^-2, respectively; the nitrogen surplus under HV could be decreased by 7.94 and 0.14 kg·hm^-2, respectively; HV could replace a maximum nitrogen application rate of 15.71% and 19.71%, respectively, which could replace conventional nitrogen application rates of 15.71% and 26.23%, respectively. 【Conclusion】 The incorporation of hairy vetch could activate soil enzyme activity, effectively enhance soil nitrogen nutrient supply capacity, delay post popcorn leaf senescence, increase the contribution rate of post flowering dry matter accumulation to grains, and improve ear traits and coordinate yield composition, which all were beneficial for increasing and stabilizing maize yield after nitrogen reduction. The input of soil nitrogen by hairy vetch was the basis for its partial replacement of chemical nitrogen fertilizer and compensation for the yield loss caused by chemical nitrogen reduction. The use of hairy vetch instead of partial chemical nitrogen fertilizer in the seasonal fallow areas of winter wheat in North China was a sustainable nitrogen reduction measure.

PLANT PROTECTION

Stem Rust Resistance Genes Identification and Evaluation of 88 Wheat Cultivars (Lines) in Heilongjiang Province

GAO Fu, WANG Rui, LIU DongJun, SUN HuiYan, WANG ZiYe, SONG WeiFu, LI TianYa

Scientia Agricultura Sinica. 2024, 57(13):  2568-2582.  doi:10.3864/j.issn.0578-1752.2024.13.006

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【Objective】 In recent years, the new races of Puccinia graminis f. sp. tritici (Pgt), Ug99, TKTTF, and TTRTF, have emerged and continuously spread. Heilongjiang Province is an area where wheat stem rust occurred frequently. The aim of this study is to analyze the resistance genes contained in wheat cultivars in this region and their resistance to predominant races of Pgt, and to provide a theoretical basis for the rational regional layout of resistance genes and the breeding of disease resistant cultivars in China. 【Method】 Based on the response types of 43 monogenic lines and 88 wheat cultivars (lines) to 7 different virulence races (21C3CTTSC, 21C3CTHQM, 34MKQSM, 34C3MTGQM, 34MKGQM, 34MTGSM, and RTGRM) of Pgt, the stem rust resistance genes contained in these 88 cultivars (lines) were postulated; Further, the presence of Sr31, Sr24, Sr25, Sr26, and Sr38 in the tested cultivars (lines) was detected by using molecular markers closely linked with these five genes. At the same time, the resistance of these cultivars (lines) to two predominant races 21C3CTHQM and 34MKGQM was evaluated at the adult-plant stage during the period of 2021-2022.【Result】 The results of gene postulation showed that 37 wheat cultivars might contain one or more of the wheat stem rust resistance genes Sr9e, Sr11, Sr17, Sr23, Sr30, Sr35, Sr37, Sr39, Sr40, and SrWld. In addition, molecular testing confirmed that 10 wheat cultivars (Kenmai 16, Longchun 204, Longfumai 8171, Longmengmai 2305, Longfumai 7166, Longchun 218, Longmai 95, Longfumai 19-92, Kechun 171286, and Longfumai 194) contained the resistance gene Sr31, but none of wheat cultivars (lines) carried Sr24, Sr25, Sr26, and Sr38. In field trials in 2021 and 2022, 86 (97.7%) and 83 (94.3%) wheat cultivars showed resistance to race 21C3CTHQM and 34MKGQM, respectively. Among them, 62 (70.5%) wheat cultivars showed good resistance to both tested races for two consecutive years, while the other cultivars (lines) showed different degrees of susceptibility to a single race.【Conclusion】 Wheat cultivars (lines) in Heilongjiang Province have good resistance to the tested races at adult-plant stage, and 37 wheat cultivars may carry one or more of the 10 resistance genes (Sr9e, Sr11, Sr17, Sr23, Sr30, Sr35, Sr37, Sr39, Sr40, and SrWld). There are 10 cultivars (lines) containing Sr31, and no wheat cultivars (lines) containing Sr24, Sr25, Sr26 and Sr38 resistance genes were detected.

The Dynamic Wetting and Spreading Behavior of Pesticide Droplet on Rice Leaf Surface

ZHANG JianTao, HUANG LuSheng, LIU GuangBin, LAN YuBin, WEN Sheng

Scientia Agricultura Sinica. 2024, 57(13):  2583-2598.  doi:10.3864/j.issn.0578-1752.2024.13.007

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【Objective】 The objective of this study is to investigate the effects of surface tension, droplet size, and leaf angle on the dynamic wetting and spreading behavior of pesticide droplets on both adaxial and abaxial surfaces of rice leaf, and to provide a basis for realizing the “reduce application and increase efficiency” of rice spraying by regulating the dynamic wetting and spreading behavior of pesticide droplets on the rice leaf surface.【Method】 A full factorial experiment was designed to investigate the dynamic wetting and spreading behavior of single droplet on both adaxial and abaxial surfaces of rice leaf in this study. The Silwet-408 solutions with surface tension of 21.4, 33.2, and 43.7 mN·m^-1 were formulated by adjusting the concentration of Silwet-408 to replace the pesticide solutions. A droplet generator was used to produce the single droplets of 532, 627, 746, 830, and 957 μm. The leaf angles were set as 40°, 65°, and 85°.【Result】 There were significant effects of surface tension, droplet size, and leaf angle on the rate of change of droplet contact angle on the rice leaf surface (P<0.05), and the overall trend of the influences on the adaxial and abaxial surfaces was basically the same; increasing leaf angle or reducing droplet size or lowering surface tension could increase the rate of change of contact angle and promote the wetting and spreading of droplet. Among them, the effect of lowering surface tension was the most significant. When the surface tension was decreased from 33.2 to 21.4 mN·m^-1, which was close to the critical surface tension of rice leaf surface, the rate of change of the contact angle (advancing contact angle and receding contact angle) increased by 7.49 and 6.22 times for the adaxial surface, and 11.13 and 7.61 times for the abaxial surface, and the wettability of the droplets was changed from relatively poor or poor (80°≤contact angle<100° or contact angle≥100°) to medium or positive (60°≤contact angle<80° or contact angle<60°) within 75 s; when the surface tension was much larger than the critical surface tension of rice leaf surface, the rate of change of contact angle increased with the increase of leaf angle, and increased with the decrease of droplet size, but the effect of leaf angle was less than that of droplet size, and droplets of almost all particle sizes still maintained relatively poor or poor wettability (80°≤contact angle<100° or contact angle≥100°) after 75 s. In addition, the analysis of droplet wetting hysteresis phenomenon showed that the roughness of the rice leaf surface was relatively small, and the droplet wetting hysteresis phenomenon was not serious. The droplet with surface tension close to the critical surface tension of rice leaf was driven by the dynamic surface tension of the solid-liquid-gas three-phase system to wet and spread on the leaf surface, and the dynamic change of the contact angle with time could be fitted by the model θ=θ_e+Aexp(-Kt); although the droplet with a surface tension much higher than the critical surface tension of rice leaf could steadily adhere to the leaf surface without the phenomenon of roll-off, it was never able to break through the pinning effect and retention resistance of the leaf surface, and could not achieve wetting and spreading.【Conclusion】 All three factors, surface tension, droplet size, and leaf angle, significantly affect the dynamic wetting and spreading behavior of pesticide droplets on the rice leaf surface. In the actual application scenarios, since the leaf angle cannot be adjusted artificially, the surface tension and droplet size of the pesticide solution can be adjusted according to the purpose of the application to regulate the dynamic wetting and spreading behavior of the droplet. The results of this study are helpful to understand the mechanism of dynamic wetting and spreading of pesticide droplet on the rice leaf surface, and can provide theoretical support and guidance for the rational selection of surface tension and droplet size in rice pesticide application scenarios.

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

Long-Term Application of Chemical Fertilizers Induces Soil Acidification and Soil Exchangeable Base Cation Loss on Paddy in Southern China

JI JianHua, LÜ ZhenZhen, LIU ShuZhen, HOU HongQian, LIU YiRen, LIU XiuMei, LI XuHua, LAN XianJin

Scientia Agricultura Sinica. 2024, 57(13):  2599-2611.  doi:10.3864/j.issn.0578-1752.2024.13.008

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【Objective】 The effects of nitrogen, phosphorus, and potassium fertilizer application on soil acidification, exchangeable aluminum production, and loss of basic ions were assessed, so as to provide the theory basis for maintaining soil health and sustainable development of farmland. 【Method】 The field experiment, established in 1984, was designed to have five treatments, that is, CK (an early rice-late rice rotation without fertilizer), PK (N deficiency), NP (K deficiency), NK (P deficiency), and NPK (balanced chemical fertilizer application), and all the treatments, except CK, were the same rate in N nutrient supply. Soil samples of 0-10 cm, 10-20 cm, 20-40 cm and 40-60 cm collected for each treatments after late rice harvest in 2016 were used to measure soil pH, exchangeable acidity, exchangeable base cation, accumulation of exchangeable base cation, pH buffer capacity, and acidification rate, etc.【Result】 After 33 years of continuous application of chemical fertilizer, the soil pH under CK, PK, NP, NK and NPK treatments decreased significantly by 0.82, 0.91, 1.13, 0.8 and 1.19 pH units compared with an initial pH of 6.5 in the 0-20 cm soil layers, respectively, which resulted in obvious acidification of cultivated soil, and the acidification rates reached 1.1, 1.22, 1.46, 1.13 and 1.58 kmol·hm^-2·a^-1, respectively. Different fertilization treatments were different from CK treatment. Compared with CK treatment, the soil pH of the 0-40 cm layer significantly decreased by 0.28-0.38 units under NP treatment and by 0.35-0.46 units under NPK treatment. The exchangeable acidity of the soil increased by 35.5%-110.0% under NP treatment and by 30.4%-120.5% in the NPK treatment, with a significant increase in exchangeable aluminum by 56.2%-157.6% and 73.7%-189.8%, respectively. The total content of exchangeable bases in the soil decreased by 6.3%-14.9% under NP treatment and by 9.9%-13.2% under NPK treatment, resulting in a decrease in base saturation by 2.9-14.9 and 2.6-15.4 percentage points, respectively. The NK treatment slightly acidified the 0-20 cm soil layer, with an increase in exchangeable acidity by 53.5%-55.0% and a decrease in base saturation by 6.0-7.1 percentage points. The PK treatment did not show significant soil acidification in the 0-60 cm layer, and there was no significant difference in the increase of exchangeable acidity and the decrease of exchangeable bases. 【Conclusion】 Long-term application of fertilizers showed significant differences in the degree of soil acidification and loss of basic ions. Among them, the long-term application of NPK and NP fertilizers intensifies the soil acidification process, with acidification depth reaching 40 cm, and there was a substantial increase in the loss of basic ions and the production of exchangeable aluminum. Preliminary estimation showed that the application of fertilizer could lower the soil pH by one unit, and the loss of exchangeable base in soil was approximately twice the increase in exchangeable acid. Long-term application of PK and NK fertilizers had a relatively smaller impact on soil acidification, with less increase in the loss of basic ions and exchangeable aluminum.

Effects of Long-Term Application of Biochar on Nutrients, Fractions of Humic in Brown Soil

WANG QingYang, CAO DianYun, WANG Di, ZHAN ZengYi, HE WanYing, SUN Qiang, CHEN WenFu, LAN Yu

Scientia Agricultura Sinica. 2024, 57(13):  2612-2622.  doi:10.3864/j.issn.0578-1752.2024.13.009

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【Objective】 The aim of this study was to investigate the effects of long-term application of biochar on nutrient content and humus component content in brown soil, so as to provide the scientific basis for evaluating the long-term effects of biochar on improving soil fertility level and regulating soil humus composition and stability.【Method】 Based on the field positioning experiment of biochar, using a randomized block design four carbon application treatments were set up: 0 (CK), 15.75 t·hm^-2 (BC1), 31.5 t·hm^-2 (BC2) and 47.25 t·hm^-2 (BC3). The soil organic carbon, available nitrogen, phosphorus, potassium nutrients, total nitrogen, phosphorus, potassium nutrients and soil fulvic acid (FA), humic acid (HA), humin (HM) content were measured after 4 and 8 years of biochar application, respectively.【Result】 The application of biochar could increase the SOC content, and the SOC content under BC1, BC2, and BC3 treatments increased by 37.35% to 72.97% than that under CK treatment. The application of biochar significantly increased the soil available potassium (AK) content; compared with CK treatment, the soil AK content of BC1, BC2 and BC3 treatments increased by 11.67-14.00 mg·kg^-1, 19.33-22.33 mg·kg^-1 and 12.33-35.33 mg·kg^-1, respectively. The application of biochar had little effect on soil total nitrogen (TN), alkali hydrolyzed nitrogen (AN), total phosphorus (TP), available phosphorus (AP) and total potassium (TK) content (except for individual treatments). The application of biochar significantly increased the content of soil HA, humus-extractable (HE), and HM. Compared with CK treatment, the soil HA content under BC1, BC2, and BC3 treatments increased by 39.68%-40.91%, 30.91%-50.79% and 34.55%-57.14%, respectively. Compared with CK treatment, the soil HA content under BC1, BC2, and BC3 treatments increased by 39.68%-40.91%, 30.91%-50.79% and 34.55%-57.14%, respectively. Compared with CK treatment, the soil HE content of BC1, BC2 and BC3 treatments increased by 18.02%-29.74%, 16.81%-30.48% and 15.92%-24.91%, respectively. Compared with CK treatment, the soil HM content of BC1, BC2, and BC3 treatments increased by 48.39%-58.94%, 13.57%-89.23% and 82.36%-105.82%, respectively. After applying biochar for 4 years, there was no significant effect on soil FA content, but after applying biochar for 8 years, the soil FA content significantly increased, with BC1, BC2, and BC3 treatments increasing by 22.01%, 30.19% and 18.24%, respectively, compared with CK treatment. In 2016, the various carbon application treatments increased soil HA/HE and HA/FA, but decreased HE/HM; but in 2020, only BC1 treatment significantly increased soil HA/FA, while BC3 treatment significantly reduced HE/HM. Through redundancy and correlation analysis, there was a significant positive correlation between SOC, TK, AK, TP and AP and the content of humic substances; There was a significant positive correlation between SOC and TK and the stability of soil humus substances, however, there was a significant negative correlation between SOC, AK, TP, and AP and the activity of soil humus substances.【Conclusion】 Long term application of biochar could improve soil nutrient status, mainly manifested in increasing SOC and AK content. Long term application of biochar could increase the content of humus components in soil, mainly manifested in increasing the content of soil HA and HM, however, the impact on the stability of soil humus would weaken over time. There was a significant positive correlation between soil nutrient content and soil humus component content and stability.

Screening of Deodorizing Bacteria and Its Application in Composting

WEI QiHang, FENG Yao, WANG XiaoXing, ZHU HongGang, FANG Zhao, LI ZhaoJun

Scientia Agricultura Sinica. 2024, 57(13):  2623-2634.  doi:10.3864/j.issn.0578-1752.2024.13.010

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【Objective】 The aim of this study was to screen new deodorizing bacteria for solving the problem of odor emission from organic fertilizer plants with chicken manure as the main raw material, and providing the theoretical basis and support for aerobic composting biological deodorization technology.【Method】 The chicken manure was used as the sample to screen deodorizing bacteria to meet multiple goals by qualitative preliminary screening, domestication and enrichment, separation and purification, quantitative re-screening, antagonism test and drug-resistance test. The strains were identified by morphological observation and 16S rDNA sequence, and the compound bacteria agent BH was prepared and applied to chicken manure composting. Three treatments were set up, including raw materials without biological agents (CK), raw materials mixed with 1% biological deodorizing agent BH (DT1), and raw materials mixed with 1% purchased biological deodorizing agent (DT2). Some indexes were detected, including temperature, pH, electroconductibility (EC), gentamicin (GM) content and NH₃ emission, to explore the effect of BH during composting.【Result】 15 strains of bacteria were screened and isolated, named as BH1-BH15, but no fungi were screened. After rescreening, it was found that strains BH2, BH5, BH9, BH11, BH12 and BH15 had higher removal rates of NH₃, which were 48.8%, 49.4%, 45.8%, 48.3%, 51.0% and 51.8%, respectively, and there was no antagonism among the strains. The study on the drug-resistance of deodorizing strains showed that strains BH11 and BH12 had strong resistance to gentamicin. BH11 was identified as Bordetella sp. and BH12 was identified as Weeksella massiliensis by morphology and molecular biology. Then strain BH11 and BH12 were made into compound bacterial agent BH, and its deodorizing effect was significantly better than that of single bacterial agent, with the removal rate of NH₃ was 65.8%. During composting, the thermophilic phase of each treatment lasted for more than 9 days, and the harmless treatment of chicken manure was achieved. At the end of composting, the pH value of each treatment was stable at 8.40, between 5.5 and 8.5, and the EC was between 2.73 and 3.43 mS·cm^-1, lower than 4 mS·cm^-1, indicated that the material could be used as plant fertilizer and met the organic fertilizer standards. It was found that the GM degradation efficiency of DT2 treatment was significantly higher than that of CK treatment, indicated the commercial bacterial agent could promote the degradation of gentamicin. In addition, the emission of NH₃ during composting mainly occurred in mesophilic phase and thermophilic phase, and the emission in thermophilic phase was higher than that in mesophilic phase. Compared with CK treatment, the bacterial agent BH significantly inhibited the emission of NH₃ in mesophilic phase, and the deodorization effect was better than that of the commercial bacterial agent. After entering thermophilic phase, the deodorization effect of commercial bacterial agent was better than that of bacterial agent BH.【Conclusion】 Two strains of drug-resistant deodorizing bacteria were screened and the removal rates of NH₃ were 48.3% (BH11) and 51.0% (BH12), respectively, and the removal rate of NH₃ by the compound bacterial agent BH was 65.8%. The application of bacterial agent BH to chicken manure composting could effectively reduce the emission of NH₃ during composting, and all indicators met the standards of composting maturity.

HORTICULTURE

Differential Analysis of Aroma Substance Content and Gene Expression in the Berry Skins of Different Grape Germplasms

XU MengYu, WANG JiaYang, WANG JiangBo, TANG Wen, CHEN YiHeng, SHANGGUAN LingFei, FANG JingGui, LU SuWen

Scientia Agricultura Sinica. 2024, 57(13):  2635-2650.  doi:10.3864/j.issn.0578-1752.2024.13.011

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【Objective】 This study aimed to explore the differences in the types, content, and related gene expression levels of volatile aroma compounds in the mature grape skins of different grape germplasms, so as to provide a reference basis for the analysis of the metabolic regulation mechanism of grape volatile aroma substances and the full utilization of aroma resources. 【Method】 In this study, GC-MS was used to identify the types and content of volatile aroma substances in the pericarp of 13 grape germplasms. The transcriptional level of the entire genome was also analyzed by using Illumina high-throughput sequencing platform.【Result】 A total of 154 volatile aroma substances were detected in the 13 grape germplasms, mainly including aldehydes, esters, terpenoids and other compounds. The germplasm with the highest content of volatile aroma substances in the skin was Kyoho, while the lowest was Cabernet Sauvignon. Esters were the most abundant volatile aroma substances in the skins of Kyoho and Petit Verdot, while aldehydes were the most abundant volatile aroma substances in the other germplasms skins. Twelve characteristic aroma substances such as hexanal, ethyl caprylate and linalool contributed more to the aroma of grape skins. The results of principal component analysis showed that there were large differences in volatile aroma substances in the skins of Kyoho, Thompson Seedless, Red Globe and other germplasm. The results of transcriptome analysis showed that the number of differentially expressed genes (DEGs) varied considerably among different germplasms. DEGs were significantly enriched in the pathways of linoleic acid metabolism, α-linolenic acid metabolism, phenylalanine metabolism, tyrosine metabolism, and monoterpene biosynthesis. The expressions of lipoxygenase (LOX), alcohol acyltransferase (AMAT), deoxyd-xylose-5-phosphate reductase (DXS), alcohol dehydrogenase (ADH) and other genes were highly correlated with the content of aroma compounds. Further analysis revealed that the transcription factors WRKY7, WRKY28, ARF4, and ARF22 were potential regulators of aroma synthesis in grapes.【Conclusion】 The content of aroma substances and the expression of related genes in the pericarp of different grape germplasm varied greatly, and the results of the joint multi-omics analysis provided a reference for further research on the synthesis mechanism of volatile aroma substances in grapes.

Cloning and Activity Analysis of U6 Promoter in Rosa chinensis Old Blush and Rosa multiflora

QIN YuQi, SU JianXin, CAO XueMin, ZHU Wan, CHENG WenHan, ZHANG Wei

Scientia Agricultura Sinica. 2024, 57(13):  2651-2661.  doi:10.3864/j.issn.0578-1752.2024.13.012

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【Background】 The U6 promoter is an important element for driving sgRNA transcription in the CRISPR/Cas9 gene editing system, and the transcriptional activity of U6 promoter in various species may be different, while endogenous U6 promoters of species may have higher efficiency than that of exogenous U6 promoters. So far, there have been only sporadic reports on the U6 promoter in genus Rosa, and no promoter with stronger transcriptional activity than Arabidopsis U6 has been obtained. 【Objective】 This study aimed to screen out the U6 promoter of Rosa plants with high transcriptional activity to optimize the CRISPR/Cas9 gene editing system, so as to lay the foundation for molecular breeding in Rosa plants, such as R. chinensis and R. multiflora in future. 【Method】 Six OBU6 promoters with the highest similarity were cloned from the genome of R. chinensis Old Blush (OB), and seven RmU6 promoters with the highest similarity were cloned from the genome of R. multiflora using conserved 102 bp U6 snRNA sequence of Arabidopsis thaliana. The fusion expression vectors of LUC and GUS reporter genes driven by OBU6 promoter and RmU6 promoter were constructed, respectively. Tobacco (Nicotiana benthamiana) leaves and tissue cultured plantlets of R. Samantha were transfected through Agrobacterium-mediated transient transformation. The transcriptional activities of promoters were determined according to the luciferase activity and GUS histochemical staining. 【Result】 Six OBU6 promoters and seven RmU6 promoters all had two essential elements that affected the transcriptional activity of U6 promoter, the USE and TATA boxes. The results of the luciferase activity and GUS histochemical staining showed that all of these U6 promoters had transcriptional activity, however, the transcriptional activity of OBU6 promoters were weaker than that of AtU6-1 promoter. Considering that an excessively long U6 promoter might weaken its transcriptional activity, OBU6-4 with relatively high transcriptional activity was selected for 5' end truncation (1 507 bp, 1 076 bp, 574 bp and 187 bp), but the truncated promoter failed to increase the transcriptional activity. And yet, RmU6-2 promoter (630 bp) was successfully identified from R. multiflora, with significantly higher transcriptional activity compared with AtU6-1 promoter. 【Conclusion】 Six OBU6 promoters were cloned from the genome of R. chinensis Old Blush and seven RmU6 promoters were cloned from the genome of R. multiflora. Finally, an RmU6-2 promoter with significantly higher transcriptional activity than AtU6-1 was obtained, which could be served as a highly promising promoter for the construction of CRISPR/Cas9 genome editing system in Rosa plants.

FOOD SCIENCE AND ENGINEERING

Effects of Semi-Solid Fermentation with Lactobacillus on the Bitterness and Active Components of Shatianyu (Citrus grandis L. Osbeck) Fruit Powder

YE JiaMin, ZHANG MingWei, LU Qi, ZHANG RuiFen, DENG Mei

Scientia Agricultura Sinica. 2024, 57(13):  2662-2673.  doi:10.3864/j.issn.0578-1752.2024.13.013

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【Objective】 The effects of semi-solid fermentation with different lactobacillus on the bitterness and active components in Shatianyu (Citrus grandis L. Osbeck) fruit powder (SFP) were explored. The aim of the present study was to establish the debitterizing technology and prepare low-bitterness SFP enriched in flavonoids and dietary fiber (DF), for providing the theoretical basis and technical support for the application of SFP in healthy food.【Method】 Shatianyu septum and pulp were dried and crushed to generate SFP, then, the bitterness of SFP and the equivalent main flavonoid glycosides (naringin and melitidin) in SPF were evaluated by sensory evaluation to clarify the bitter substance base. Lactobacillus plantarum GIM 1.1516 (LP1.1516), Lactobacillus casei GIM 1.411 (LC1.411), Lactobacillus acidophilus GIM 1.731 (LA1.731) and Lactobacillus rhamnosus GIM 1.325 (LR1.325) were used to ferment Shatianyu homogenate. During fermentation, the reducing sugar content, pH and acidity were dynamically monitored, the naringin and melitidin degradation rate were analyzed by HPLC, and the bitterness of Shatianyu homogenate after fermentation was also evaluated to determine the optimal fermentation conditions. Then, the fermented SFP was prepared, and its bitterness, together with total phenolic, total flavonoids, total DF (TDF), soluble DF (SDF), and insoluble DF (IDF) content were determined, respectively. Furthermore, the oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), ABTS free radical scavenging ability, together with water retention (WRC), water swelling (WSC), and oil adsorption capacity (OAC) of fermented SFP were determined, respectively.【Result】 SFP exhibited a more bitter taste, which was mainly caused by the co-flavor of naringin and melitidin, the main flavonoid components in SFP. During the first 12 h-fermentation, LP1.1516, LC1.411, LA1.731 and LR1.325 could use the reducing sugar in the homogenate and produce acids, while during 12-24 h-fermentation, only LR1.325 could still use the reducing sugar and produce acids. Shatianyu homogenate fermented by LR1.325 for 12 h showed the highest total degradation rates of naringin and melitidin (41.1%), also showed the highest sensory bitter removal rates (66.0%). Compared with unfermented SFP, the taste of SFP prepared by 12 h-LR1.325 fermentation changed from bitterer to between little bitter and slightly bitter, with a sensory bitter removal rate of 60.8%. Furthermore, the later showed higher total flavonoid content (increasing 19.8%), lower IDF (decreasing 9.6%), and higher SDF content (increasing 19.6%). Also, SFP prepared by 12 h-LR1.325 fermentation showed higher ORAC antioxidant activity (increasing 24.1%), higher water retention (increasing 20.8%) and water swelling capacity (increasing 32.0%).【Conclusion】 12 h-LR1.325 fermentation of Shatianyu homogenate consisting of septum and pulp generates low-bitterness SFP enriched in flavonoids and DF, which could be used in the development of related functional foods.

ANIMAL SCIENCE·VETERINARY SCIENCE

Cloning, Expression Characterization, and Functional Analysis of the Snail1 in Qinchuan Cattle and Its Impact on Proliferation of Bovine Adipocytes

ZHU BingLin, YU JiaLi, CHEN JiaYue, TIAN Yuan, WAN Yuan, LIU ChenYang, WANG XiaoYu, WANG MiaoLi, CHENG Gong

Scientia Agricultura Sinica. 2024, 57(13):  2674-2686.  doi:10.3864/j.issn.0578-1752.2024.13.014

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【Objective】 The deposition of intramuscular fat in beef cattle is closely associated with the flavor, juiciness, and tenderness of beef, characterized by the proliferation (increased in number) and differentiation (lipid deposition) of fat cells, which is regulated by multiple genes. The previous studies have indicated that Snail1 is involved in muscle development and lipid homeostasis in mice; however, its role in bovine adipogenesis remains unknown, which warrants further investigation. 【Method】 Snail1 gene CDS region sequence was cloned from Qinchuan cattle, Snail1 gene temporal and spatial expression profile was constructed, and its functional structure and target gene were predicted by bioinformatics software. Subsequently, the impact of Snail1 on the proliferation of bovine adipocytes was explored through RNAi interference, along with CCK8, EdU, cell flow cytometry, and RT-qPCR methods. 【Result】 Compared with the sequence published by NCBI, the Snail1 gene in Qinchuan cattle had two synonymous mutations. The Snail1 gene exhibited higher abundance expression in newborn cattle lung, perirenal fat, and small intestine. However, in the adult cattle, the expression level of the Snail1 gene was the highest in perirenal fat tissue, followed by longissimus dorsi muscle, and lowest in lung tissue. Bioinformatics analysis revealed a 651 bp CpG island in the Snail1 promoter region, along with binding sites for transcription factors related to fat formation, such as C/EBP and PPARα. CKⅠ (Ser92/96), CKⅡ (Ser25/119, Thr89), CDK1 (Ser13/104/112/119/143/221/183/214), CDK5 (Ser105/107), and other cell cycle related kinases could participate in the Snail1 phosphorylation of proteins. By extracting promoter regions of bovine annotated genes, predicting target genes and constructing KEGG dynamic networks, it was found that lipid formation related signaling pathways, such as MAPK, PI3K-Akt and mTOR, were potential node signaling pathways involving in Snail1’s involvement in adipogenesis. Further functional studies through RNAi interference demonstrated that downregulation of the Snail1 gene promoted the proliferation of bovine preadipocytes, increased the proportion of cells in the S phase (P<0.01), and facilitated G1/S cell cycle transition. RT-qPCR and Western blot analysis indicated that interfering with the Snail1 gene significantly upregulated the expression of proliferation- regulating genes, including CCNB1, CCND2, CDK2, and CDK4 (P<0.05), at both the genes and protein levels. 【Conclusion】 The expression of Snail1 gene was relatively high in neonatal and adult perirenal fat and longissimus dorsi muscle. Interfering with the Snail1 gene promoted the proliferation of bovine preadipocytes and the G1/S cell cycle transition, as well as the expression of CCNB1, CCND2, CDK2, CDK4, and other proliferation-related genes. Cell cycle-related kinases, including CKⅠ, CKⅡ, and CDK1/5, could participate in the regulation of cell proliferation through the phosphorylation modification of Snail1 protein. Furthermore, the MAPK, PI3K-Akt, and mTOR pathways were potential node signaling pathways influenced by Snail1. This study established a foundation for further exploration of the mechanism by which Snail1 participated in bovine fat formation.

Research Progress of ApiAP2 Transcription Factors in Regulating the Growth and Development of Toxoplasma gondii

HU DanDan, LUO RunQi, LIANG RuiYing, WANG Lei, LIANG Lin, SI HongBin, DING JiaBo, TANG XinMing

Scientia Agricultura Sinica. 2024, 57(13):  2687-2697.  doi:10.3864/j.issn.0578-1752.2024.13.015

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Toxoplasmosis is one of the most serious zoonotic parasitic diseases in the world. WHO data show that the positive rate of Toxoplasma gondii antibody in the global population is 25%-50%. The vertical transmission caused by T. gondii infection in pregnant women seriously endangers the health of fetuses and infants, and T. gondii infection is also the main cause of death in immunocompromised patients. In livestock production, toxoplasmosis causes abortion and stillbirth in pregnant animals, which is seriously harmful. T. gondii is an obligate intracellular protozoan parasite that belongs to the apicomplexan protozoa, and its life cycle is complex, including asexual reproduction in intermediate hosts (humans and other warm-blooded animals) and sexual reproduction in terminal hosts (cats). In order to complete the complex life cycle of T. gondii in the intermediate and terminal hosts, its transformation and growth at different developmental stages require strict and accurate gene regulation. Therefore, revealing the regulatory mechanism of T. gondii growth and development is of great significance for the development of therapeutic drugs and vaccines. ApiAP2 transcription factor is a kind of protein family with AP2 domain and strong regulatory function, which plays a core regulatory role in the growth and development of parasites such as Plasmodium and T. gondii. It is a breakthrough to elucidate the regulation mechanism of development and transformation of T. gondii at different life cycle stages. As a model organism for the study of apicomplexan, 67 transcription factors containing AP2 domain have been annotated in T. gondii genome. Some of these transcription factors play crucial regulatory roles in the growth, development, and transformation processes of T. gondii throughout its life cycle. However, the biological functions of most AP2 transcription factors are unknown, especially the research on the development and regulation of sexual reproduction stage is relatively scarce. In this paper, the research methods, biological functions, regulatory interaction genes and networks of AP2 transcription factors of T. gondii have been systematically reviewed. The aim was to explore the genes or molecules that play a core regulatory role in the important nodes of T. gondii growth and development. The regulatory mechanism of different growth and development stages of T. gondii complex life cycle was preliminarily outlined at the micro molecular level, and its role and potential in the development of new drugs and vaccines were prospected. To provide new ideas and entry points for the prevention and control of animal toxoplasmosis and block the source of human toxoplasmosis infection, we should practice the concept of “disease prevention in animals and front-line defense” and strive to achieve “one world, one health” for everyone’s well-being.

PERSPECTIVE

China Urgently Needs to Transform from Mainland Agriculture to Cross-Sea Agriculture

REN JiZhou, JIAO Hong, YANG RuiXue, XU Gang, ZHAO An

Scientia Agricultura Sinica. 2024, 57(13):  2698-2702.  doi:10.3864/j.issn.0578-1752.2024.13.016

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Relying on its special geographical advantages, China has established a continental agricultural country. Chinese land area was nearly 10 million square kilometers, while its population had never exceeded 80 million from the Qin and Han Dynasties to the Ming Dynasty, so known as “a vast territory with abundant resources”. However, during the Qianlong period in the 19th century, its population suddenly increased to 400 million, and the land area was already insufficient. In the early days of the founding of the People’s Republic of China, the population reached 650 million, and further reached 1.41 billion in 2020 based on The Seventh Census. The per capita water and soil resources are only about 1/3 to 1/4 of the world level, which making it a resource-poor country. It is not easy to obtain enough food and even more difficult to achieve a moderately prosperous lifestyle relying solely on domestic resources. The shortage of water and soil resources is an urgent problem despite the huge potential for technological innovation. In nearly a century and a half, the historical tragedy of the collision between China’s agricultural civilization and the world’s industrial civilization has reminded us to break through the mentality of “domestic is the world” fostered by land-based agriculture and to set sail to the sea. China needs to transform from “self-sufficient” mainland agriculture to “shared supply and common sustenance” cross-sea agriculture, establishing itself as the main base, utilizing the world’s agricultural resources, and building world agriculture. It is an urgent need to set up a long-term national policy in order to transform mainland agriculture into cross-sea agriculture, to build a global agricultural database in order to respond to the demand of the international food market, to adjust accurately and timely the domestic agricultural structure in order to reduce international trade risks, and to make a global strategic deployment as soon as possible for China’s agriculture.