10 February 2023, Volume 37 Issue 2
    

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    Induced Mutations for Plant Breeding·Agricultural Biotechnology
  • FENG Yuan, LIU Yefang, KOU Xiaohong, XUE Zhaohui
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 217-229. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0217
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    NAC transcription factors are involved in plant growth and development, fruit pigment accumulation, cell wall morphogenesis, and plant senescence. To systematically study the precise function of SNAC4SlNAC48, Gene ID: 101247735) and SNAC9SlNAC19, Gene ID: 101248665) in the tomato ripening and aging, we designed SNAC4/9 knockout targets and constructed a single guide RNA (sgRNA) expression cassette through Overlapping Polymerase Chain Reaction (Overlapping PCR) method. Single or multiple sgRNA expression cassettes were assembled into pYLCRISPR/Cas9 vector by the Golden Gate cloning method. PCR and sequencing results showed that the SNAC4/9 knockout vector was successfully constructed. The genes encoding Cas9 protein and sgRNA were introduced into Micro-Tom tomato cells by the Agrobacterium transformation method, and the target and off-target of positive seedlings were detected. The results showed that tomatoes were mutated successfully without off-target. The identification of T1 mutants further demonstrated that CRISPR/Cas9 gene editing targets in tomato could be inherited stably between generations. Compared with wild type, SNAC4/9 knockout fruits had less pigment accumulation and delayed ripening, suggesting that SNAC4/9 played an essential role in tomato fruit ripening. This study provided genetic material and an experimental basis for further exploring the mechanism of SNAC4/9 regulating pigment metabolism and cell wall metabolism in tomatoes.

  • FANG Yuanpeng, WEI Jianming, LI Yunzhou
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 230-240. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0230
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    Tomato brown rugose fruit virus (ToBRFV) belongs to the genus Tobamovirus in the Virgaviridae family and is classified as a quarantine virus in China, posing a serious threat to the safety of tomato production. Signal transduction system plays an important role in plant disease resistance, and phospholipid signaling is crucial in the process of extracellular resistance signal transduction. Phospholipase C (PLC) is critical in the transmembrane signaling of the phospholipid signaling system. In this study, we first identified 10 tomato PLC genes based on bioinformatics, including seven phosphati-dylinositol-specific phospholipase C (PI-PLC) and three non-specific phospholipase C (Non-specifc -PLC, NPC), the seven PI-PLC PLC proteins have three core structural domains (PLC_X c, PLC_Y c, C2) and an EF_hand-like structural domain, and the three NPC proteins only have a phosphoesterase structural domain. 10 tomato PLC proteins can be divided into seven branches according to their structural similarity, namely NPC1, NPC2, NPC6, PI-PLC2, PI-PLC3, PI-PLC4 and PI-PLC6. In addition, the 10 tomato PLC secondary structures were similar in proportion, but there were significant differences in tertiary structures. Co-linearity analysis showed that there were 3, 12 and 16 pairs of co-linear relationships between the distribution of tomato PLC genes and rice, Arabidopsis and Raymond-type cotton PLC genes. Finally, to clarify which tomato PLCs are involved in the plant defense response against ToBRFV, this study examined the relative expression levels of the PLC gene family after ToBRFV inoculation by transcriptome sequencing, and the results showed that SlNPC1SlNPC6 and SlPLC4 were expressed at higher levels in the ToBRFV-inoculated samples, while the expression levels of other PLC genes were reduced after ToBRFV inoculation. This study lays the technical and theoretical foundation for tomato ToBRFV resistance research and breeding.

  • DING Yanqing, XU Jianxia, WANG Can, ZHOU Lengbo, ZHANG Guobing, ZHAO Qiang, SHAO Mingbo, ZHANG Liyi
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 241-250. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0241
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    In order to explore the genetic basis of grain brewing traits of liquor-making sorghum, in this study, 205 recombinant inbred lines (BTx623×HYZ) were genotyped using super genotyping based on sequencing (Super GBS) technology, and a genetic map including 1910 single nucleotide polymorphism (SNP) with a marker interval of 0.47 cM were constructed. Based on the phenotype data in four environments and inclusive composite interval mapping (ICIM) method, 9, 7, 11, 5 and 3 quantitative trait locus (QTLs) related to total starch content, amylopectin content, tannin content, grain hardness and grain color respectively were detected on chromosome 1 to 9. Totally, 35 unique QTLs were identified, of which 15 were consistent with the previous studies. Three important genetic regions on chromosome 1 (66.30-71.55 Mb), chromosome 4 (54.00-62.3 Mb) and chromosome 6 (54.59-57.57 Mb) were detected in multiple traits and environments. Among which including Tan1 and Y1 genes, as well as 6 candidate genes involving in starch and flavonoid biosynthesis pathway, such as β-amylase, flavanone 3-hydroxylase and bHLH transcription factors. Our results not only lay the foundation for further cloning genes related to brewing traits, but also provide a basis for accelerating the breeding of new varieties for liquor-sorghum using marker-assisted breeding.

  • CHEN Na, SHAO Qin
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 251-261. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0251
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    To investigate the role of NAC transcription factor in response to biotic stress in tomato (Solanum lycopersium), SlNAP2 was cloned from tomato using the RT-PCR technique. The full-length of SlNAP2 cDNA was 1 227 bp, containing 142 bp 5' and 257 bp 3' non-coding regions, the open reading frame of 828 bp encodes 275 amino acids. The matching protein contained a single NAM domain. Homologous sequence alignment and phylogenetic tree analysis revealed that SlNAP2 had the most homology with Solanum pennellii, followed by Solanum tuberosum. According to the bioinformatics study, the molecular weight of SlNAP2 is 31 877.23 Da, the theoretical isoelectric point was 8.56, and the encoded protein has no transmembrane structure. SlNAP2 gene expression was tissue-specific, with the maximum expression level in tomato stem. Functional analysis revealed that SlNAP2 participated in response to bacterial wilt stress, salicylic acid and jasmonic acid stimulation. Silencing SlNAP2 via virus induced gene silencing (VIGS) reduced the resistance of tomato to bacterial wilt, indicating that SlNAP2 plays a negative role in tomato resistance to bacterial wilt. This study served as a foundation for further research into the significance of the SlNAP2 gene in tomato bacterial wilt resistance.

  • HU Hui, GAO Ruoyu, LI Zhixin, XU Junying, YANG Longwei, TIAN Yu, QIU Xianjin, XU Jianlong
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 262-273. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0262
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    Ratoon rice has high appearance quality. To understand the genetic basis of appearance quality of ratoon rice, two sets of reciprocal introgression lines derived from Xian variety Minghui 63, and Geng variety 02428 were evaluated for appearance quality in both first and ratoon seasons at Jingzhou for two years. The results indicated that Minghui 63 had significantly higher appearance quality than 02428 in both the first and ratoon seasons,and all appearance quality traits showed continuous distributions in the two sets of populations. There was significantly positive correlation between first and ratoon seasons within one trait, and there were significant correlations among different traits in the first or ratoon seasons. Grain width had the larget effect on appearance quality. Combined with 4 568 Bin genotyping, a total of 57 QTLs were identified in two years and distributed on all 12 chromosomes. Among them, twenty-five QTLs were simultaneously identified in both years, and seventeen QTLs were stably expressed in both backgrounds. Moreover, regions of 16.28~17.33 Mb on chromosome 3, 3.35~4.28 Mb on chromosome 5, 24.68~25.46 Mb of chromosome 7 and 6.19~6.97 Mb on chromosome 11 had pleiotropic effects on more than four traits, the alleles carried by Minghui 63 at all 4 regions could increase appearance quality. Finally, the region of 6.19~6.97 Mb on chromosome 11 for percentage of endosperm with chalkiness and degree of endosperm chalkiness in the first season was validated by an F2 population. Our results provide genetic basis for improving appearance quality of ratoon rice by molecular breeding.

  • ZHOU Bingyue, YUAN Jianlong, ZHANG Yumei, LI Li, LIANG Lijuan, XIA Lulu, ZHANG Feng
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 274-289. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0274
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    The fertility, adaptability and stability of potato are important indicators for potato popularization and application. Because of the differences in the performance of different characters by various environmental factors, it is necessary to screen and identify varieties (lines) in different ecological zones. In this study, the yield and plant traits of advance lines and main cultivars were analyzed by GGE (Genotype + genotypes and environment interactions) biplot, and the potato varieties (lines) with wide adaptability, high yield, stability and suitable for mechanization were selected. The effect of genotype-environment interactions on yield traits and plant traits were detailed, which provide a reference for potato breeding and selection of varieties in different ecological regions. Twenty two advance lines and thirteen main cultivars were selected to plant in three pilot sites in Weiyuan County, Anding District and Yongchang County of Gansu province from 2020 to 2021. Eight plant traits such as number of main stems and nine yield traits such as plot yield were measured. Analysis of variance was applied for analysis of significance. GGE biplot was used for analysis of adaptability, productivity and stability. Concurrently, the distinctiveness and representativeness of pilot sites were evaluated. Variance analysis showed that the effect of genotype on natural and absolute plant height were significant, and the sum of squares of genotype effect and the ratio to total variance was 66.63% and 56.56% respectively. The effect of genotype-environment interactions on peduncle length and yield of large tuber per plant were significant, and the sum of squares of interactions effect and the ratio to total variance was 27.86% and 27.05% respectively. The effect of environment on number of branches and yield of tuber per plant were significant, and the sum of squares of environment effect and the ratio to total variance was 55.82% and 25.52%. GGE analysis showed that yield and stability of varieties (lines) were generally consistent in different years. The genotypes with highest yield were G1, G33 and G10 in Weiyuan County, Anding District and Yongchang County. G32 was the most stable in the three pilot sites. Yongchang County had the strongest distinctiveness among varieties (lines) in 2020. Anding District had the strongest representativeness to target environment in 2021. GGE biplot can intuitively showed the results of varieties (lines) test in three sites between 2020 and 2021, and screen and evaluate varieties (lines) were G33, G12 and G3 were high yield, stable and suitable for mechanization, while G28, G26 and G23 were high yield, unstable and unsuitable for mechanization. The genotype-environment interactions had the greatest influence on peduncle length and yield of large tuber per plant. Combined with distinctiveness and representativeness, Weiyuan county was the best pilot sites in this experiment. According to the adaptability and stability of varieties (lines), the varieties (lines) suitable for planting in different ecological regions with high yield, stability and suitable for mechanization were selected, which will provide reference and theoretical basis for the subsequent registration and promotion of varieties.

  • LU Maoang, PENG Xiaoai, LI Min, SONG Youhong, HE Xianfang, ZHU Yulei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 290-297. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0290
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    In order to screen suitable identification indexes of wheat grain uniformity, and establish an evaluation model of wheat grain uniformity, 150 wheat materials were planted in two locations, the grain morphological indexes such as grain length (X1), grain width (X2), diameter (X3), roundness (X4), grain length width ratio (X5), grain perimeter (X6), grain surface area (X7) and thousand grain weight (X8) were tested. In the present study, the diversity statistics, correlation analysis, cluster analysis, principal component analysis and membership function method were used to comprehensively assess the wheat grain uniformity. The results showed that the variation of each index under two locations ranged from 3.56% to 9.88%, the wheat accessions were clustered into four groups. Two independent comprehensive components were obtained from eight single indexes by a principal component analysis, and their contribution rates were 61.330% and 36.448% respectively, which reflected 97.778% of all indexes information. The comprehensive evaluation values (D) of grain uniformity were calculated from the membership function methods. In this way, the 150 accessions were classified into 4 uniformity types, including 12 ultra-high uniformity materials, 80 high uniformity materials, 56 medium uniformity materials and 2 low uniformity materials. A mathematical evaluation model of wheat seed uniformity was developed by stepwise regression equation: VP=-2.787+0.294 X2+0.007 X8 +0.246 X3+0.869 X4-0.178 X5+0.026 X6+0.019 X7R2=0.999). The optimum regression equation established to predict the uniformity of the accessions showed a highly significant positive correlation (P<0.01) between the predicted value (VP) and the D value. This study shows that grain length can be ignored in screening for high uniformity materials, with emphasis on grain width, diameter, roundness, grain perimeter, grain area and thousand grain weight. This study not only clarifies the influencing factors on wheat seed uniformity, but also provides a new evaluation method of wheat seed uniformity. This study provides a theoretical support for subsequent breeders to select materials with high uniformity.

  • Induced Mutations for Plant Breeding·Agricultural Biotechnology
  • LIU Xiaona, ZHANG Lihua, LI Hongjun
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 298-305. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0298
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    In order to understand patent protection and technical development of crop irradiation mutation breeding technology in China, this paper constructs a retrieval method to obtain patent documents, and then uses the method of eliminating irrelevant documents and indexing documents to statistically analyze patent documents of crop irradiation mutation in China. Analysis shows that the number of applications in this field in China is still on the rise, but the grant rate is low, which is about 30%. The largest number of applications came from Anhui Province, accounting for 11% of the total. The proportion of foreign applicants is only 8%. The specific variety with the most application of irradiation mutation patents is rice, accounting for about 12%. Seeds are still commonly used as irradiation sites, γ-ray is the most widely used radiation source, and most of the patents are related to irradiation mutagenesis for mutants. By analyzing the key patented technologies, and putting forward suggestions on matters that worth noticing in patent application of this field, this paper provides a useful reference for researchers to development technology and protect intellectual property.

  • Food Irradiation·Food Science
  • MA Yanli, SONG Tingting, WEI Chunyan, XU Lihong, WANG Shulin, LI Chunlin, NIE Jing, YUAN Yuwei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 306-313. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0306
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    In order to explore the feasibility of stable isotope in the traceability of Auricularia auricula, 74 representative Auricularia auricula samples were collected from Northeast China (Heilongjiang, Jilin, Liaoning) and Zhejiang, and 11 representative Auricularia auricula samples were collected from Xingjiang as external verification. δ13C value, δ15N value, δ2H value and δ18O value were analyzed by elemental analysis-stable isotope ratio mass spectrometer, and the traceability of Auricularia auricula was determined by chemometrics. The results showed that the range of δ13C was between -24.5‰ and -22.7‰, δ15N was between -0.9‰ and 3.1‰, δ2H was between -62.2‰ and -34.6‰, and δ18O was between 15.9‰ and 19.4‰ for samples from Northeast China(Heilongjiang, Jilin, Liaoning).And the range of δ13C was from -26.2‰ to -24.5‰, δ15N was from -0.9‰ to 1.2‰, δ2H was from -24.9‰ to -9.0‰, and δ18O was from 19.9‰ to 22.2‰ for samples from Zhejiang. There was no significant difference in δ15N between the two habitats (P>0.05), while δ13C, δ2H and δ18O showed significant differences (P<0.05). Partial least square discriminant analysis showed that the overall discriminant rate of the Northeast China(Heilongjiang, Jilin, Liaoning)Auricularia auricula was 98.4% for the training set of discriminant model, and the overall rate was 95.2% for the verification set. The overall discriminant rate of Zhejiang Auricularia auricula was 100% for the discriminant model training set,and the overall discriminant rate of verification set was also 100%, which had good discriminant accuracy. To sum up, stable isotope analysis combined with chemometrics model could well distinguish Auricularia auricula from Northeast China (Heilongjiang, Jilin, Liaoning) and Zhejiang Province, and can trace the origin of different Auricularia auricula, which has certain application value.

  • LU Wenjing, CHEN Di, YE Qin, ZHANG Cen, XIAO Chaogeng, SHEN Zheyi
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 314-320. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0314
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    To investigate the effects of proline and KCl on the quality of sea bass, the sea bass was marinated by different salting combinations (NaCl, NaCl+proline, NaCl+KCl, NaCl+KCl+proline) at 4 ℃. After of salting 18 h and 36 h or air-drying 3 h and 6 h, pH, chromatic aberration, moisture content, salt content, thiobarbituric acid (TBA) value, texture, microstructure, and indices of sensory evaluation of sea bass were determined. The results depicted that the pH value of cured sea bass showed a decreasing trend followed by an increasing trend, the water content of all samples illustrated a decreasing trend, while the salt content described an upward trend. Notably, the lowest TBA value of 1.4157 mg·kg-1 was measured after marinating sea bass with NaCl+proline. The results through transmission electron microscopy (TEM) showed that the addition of proline made the sea bass tissue tighter and better-dissolved myosin. The salt reduction formulation of NaCl+KCl+proline combined with the dry marinade to marinate sea bass resulted in higher hardness and chewiness and effective inhibition of fat oxidation, so that myosin had better agglutination and better flavor, and the quality of sea bass was improved. These results can provide technical support for industrialized salting and production of sea bass.

  • TAO Shijie, SONG Yandong, WU Qinjiao, ZHOU Yufeng, YANG Jie
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 321-328. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0321
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    In order to study the effect of altitudes and bamboo ages on the essential oils quality of Acidosasa edulis leaves, the chemical composition of the essential oils of new and old leaves at different altitudes was determined by gas chromatography-mass spectrometry (GC-MS). The disk-diffusion and DPPH method were used to measure the antibacterial and antioxidant activity of bamboo leaves essential oils. The results showed that the composition of the essential oils of Acidosasa edulis leaves was dominated by alkanes, alcohols, terpenoids and aromatic compounds.The medium to high altitudes were good for the synthesis and accumulation of terpenoids. The content of alcohols, terpenoids and aromatic compounds in the new leaves essential oils was higher than that of old. Bamboo leaves essential oils showed inhibitory effects on both Fusarium and Botryosphaeria dothidea, the higher the altitudes the better the inhibitory effects, and the new leaves essential oils showed stronger inhibitory effects than old. Bamboo leaves essential oils showed high antioxidant activity.The antioxidant capacity of the essential oils was stronger at low altitudes than at middle and high altitudes, and the new leaves was stronger than old. This study results provides data support and new utilization directions for the development of Acidosasa edulis leaves resources.

  • TIAN Lipeng, WAN Qianggui, WANG Ting, LI Chun'ai, CAI Meng, PU Lumei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 329-337. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0329
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    In order to explore the inhibitory effects of olive leaf extracts on B.cinerea and its mechanism, the olive extracts were obtained by solvent extraction, which were then were treated with B.cinerea, Colony diameter and mycelial biomass were detected to explore the inhibitory effect of the extracts on the growth of B.cinerea. The effects of olive extracts on cell membrane integrity were investigated by measuring relative conductivity, nucleic acid and protein leakage. The effects of extracts on cell membrane permeability and ROS metabolism of B.cinerea were studied by propidium iodide (PI) staining and plate culture system. The results showed that B.cinerea was highly sensitive to 90% ethanol extract, and the colony diameter and mycelial biomass of treated B.cinerea were 20% and 8.91% of the control. Extract treatment also significantly enhanced the levels of membrane conductivity, nucleic acid and protein leakage, and improved the content of O2·- and H2O2 in the spores. The activities of enzymes related to ROS metabolism including NOX, CAT and POD were increased, and the activities of SOD, GR and APX decreased significantly. It was concluded that the cell membrane of B.cinerea treated with the extracts was damaged, resulting in the leakage of content and the accumulation of ROS in cells, which aggravated the occurrence of membrane lipid peroxidation, increased the content of MDA, and further inhibited the growth of cells. This study provides a possible way to utilize olive leaf extracts in the fruit and vegetable fungicides.

  • CAO Ronglu, CHEN Xiao’e, SONG Shijun, DONG Ruijuan, ZHANG Fang, MA Yuqiao, WANG Xiaoyan
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 338-345. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0338
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    In order to solve the increasingly prominent problem of adulteration in the fish oil raw material market, this study used gas chromatography to determine the fatty acid composition of sardine and five kinds of potentially adulterated marine fish. The variation pattern of each fatty acid in fish oil was investigated by examining its fatty acid composition and using principal component analysis (PCA) to screen out possible adulterated marine fish oil. The adulteration identification model of sardine oil was developed by using partial least squares discriminant analysis (PLS-DA) and multiple stepwise linear regression analysis. The results showed that the fatty acid compositions of five types of fish oil and sardine oil were different in varing degrees, among which anchovy oil and sardine oil were the closest. The total contribution of PCA analysis was 89.8% indicated that the analysis results were representative and anchoug oil could be proposed as adulterated oil of sardine. PLS-DA analysis showed that six characteristic fatty acids of sardine adulterated oil were eicosapentenoic acid (EPA) ,docosahexenoic acid (DHA), palmitic acid, palmitoleic acid, docosenoic acid and oleic acid, respectively. The correlation coefficients R2 of the sardine oil aduteration identification models were 0.927, 0.963, 0.987 and 0.991, and the average errors of the models were 2.20%, 1.38%, 1.19% and 0.83%, respectively, indicating the model had good accuracy. Therefore, this model can achieve quantitative identification of anchovy oil adulterated with fake sardine oil, and has certain feasibility in identifying adulterated sardine oil.

  • CHEN Qingwei, ZHANG Weiren, LI Qinglong, LI Ronghua, MU Changkao, WANG Chunlin, SONG Weiwei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 346-354. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0346
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    In order to investigate the effects of testosterone propionate on the composition of amino acids and fatty acids of swimming crab (Portunus trituberculatus), the juvenile crab of stage I was randomly divided into control group (no testosterone propionate (TP) in the feed), TP 200 group (200 mg·kg-1 TP in the feed) and TP 400 group (400 mg·kg-1 TP in the feed). After rearing for 90 days, the contents of TP, testosterone (T) and estradiol (E2) in serum of swimming crab were determined by liquid chromatography-mass spectrometry (LC-MS); the growth index and survival rate were recorded; basic nutrient composition, amino acid and fatty acid composition of muscle and hepatopancreas were analyzed for comparison. The results showed that the serum of P. trituberculatus contained a high content of sex hormones, indicating that it has a strong ability to enrich sex hormones. TP had no significant effect on the hepatopancreatic index or molting frequency of P. trituberculatus, but significantly reduced the survival rate (P<0.05). The weight gain rate, specific growth rate and full carapace width of P. trituberculatus in TP 200 group were significantly higher than those in control group and TP 400 group (P<0.05). The contents of total amino acids and essential amino acids in muscle and hepatopancreas of TP 200 and TP 400 groups were not significantly different from those of the control group. However, the content of arginine decreased significantly; there was no significant difference in the composition and content of fatty acids in the muscle between TP 200/400 groups and control group, but the content of some fatty acids in hepatopancreas, especially highly unsaturated fatty acids (C20∶2, EPA and DHA) decreased significantly in the experimental groups, which may be related to the acceleration of fatty acid oxidation and energy supply by sex hormones. These results would provide basic data for the study of the effects of exogenous sex hormones on the metabolism of amino acids and fatty acids in invertebrates.

  • ZHANG Chi, LI Shuhui, ZHOU Hongyuan, GUO Ting, ZHANG Yuhao, MA Liang
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 355-361. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0355
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    To understand the protective effect of quercetin on α-ZOL-induced damage to Caco-2 cells, the effects of quercetin on the oxidative damage and mitochondrial damage were analyzed using reactive oxygen species, mitochondrial membrane potential, malondialdehyde, superoxide dismutase activity and ATP content. AO fluorescence staining and flow cytometry were used to detect apoptosis. The relative expression levels of Nrf2, Bcl-2, Caspase-3 protein were determined by western blot. Results indicated that the degree of oxidative damage of Caco-2 cells increased with the α-ZOL concentration. Compared with model group, quercetin reduced the oxidative stress of α-ZOL on Caco-2 cells by decreasing the level of reactive oxygen species and malondialdehyde, as well as increasing the activity of antioxidant enzymes. The mitochondrial function can be improved by increasing ATP levels and mitochondrial membrane potential. Additionally, by up-regulating the expression of Nrf2 and Bcl-2, as well as down-regulating the expression of Caspase-3, quercetin can reduce apoptosis induced by α-ZOL (P<0.05). The results could be used to prevent and control the toxicity of α-ZOL.

  • Isotope Tracer Technique·Ecology and Environment·Physiology
  • WU Jiale, QI Wenle, WU Jingjing, QUE Renwei, ZENG Yongjun, PAN Xiaohua, XIE Xiaobing
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 362-369. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0362
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    To improve grain yield and quality simultaneously, this study aimed to find the best timing for panicle fertilizer application of high-quality late indica rice (Oryza sativa L.subsp. xian). Three panicle developmental stages (D1: elongation phase of the fourth leaf from the last; D2: elongation phase of the third leaf from the last; D3: elongation phase of the penultimate leaf) were chosen to understand the effects of fertilization timing on yield, nitrogen accumulation and eating quality. The 15N tracing technique was used in field micro-plot experiments with two rice varieties (YLY911 and YXYLS). The grain yield of both varieties increased at first and then decreased during the sequential panicle fertilizer applications and D2 fertilization produced the highest yield with a significant increase of 4.30%~6.39% compared to D1 and D3. Total grain nitrogen accumulation, and 15N labeled fertilizer nitrogen accumulation and its ratio to total grain nitrogen accumulation, 15N labeled fertilizer nitrogen recovery rate, and nitrogen harvest index of 15N labeled fertilizer showed an increasing trend towards the late fertilization. In comparison to D1, significant increases of 19.16%~21.26%, 32.54%~50.75%, 0.91~2.49, 12.42~23.34 and 5.18~8.31 percentage points were observed for these parameters in D2 and D3. There was no significant difference between D2 and D3 in YXYLS. The grain protein content and setback value increased, while the gel consistency, peak viscosity and breakdown value decreased as the panicle fertilization delays and therefore eating quality was reduced, but the breakdown and setback values were not different between D1 and D2 in YXYLS. Moreover, the yield, grain nitrogen accumulation, 15N labeled fertilizer nitrogen recovery rate, and nitrogen harvest index of 15N labeled fertilizer of YLY911 were significantly higher than those of YXYLS. The response of YLY911 to late panicle fertilization was greater than YXYLS. In conclusion, applying panicle fertilizer at D2 stage could synergistically achieve higher yield, better quality and higher nitrogen use efficiency in high-quality late indica rice. The results of this study provide a theoretical basis for optimizing the application period of panicle fertilizer of high-quality late indica rice.

  • QIAO Meixia, YANG Zhenping, YAN Xia, MA Yanling, LIU Yutao, GAO Zhiqiang, CHEN Jie
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 370-378. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0370
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    In order to explore the effects of exogenous actinobacteria on the seed germination and plant growth of foxtail millet at jointing stage, Streptomyces albidoflavus T4 was selected as the tested strain. A petri dish experiment was conducted to analyze the effects of the undiluted, 10-, 100-, and 1 000-times diluted culture filtrates of T4 on the seed germination of foxtail millet, with the sterile water was considered as control; a pot experiment was conducted to investigate the effects of the T4 living inoculum on the plant growth and rhizosphere microorganisms of foxtail millet, with the non-inoculum treatment as control. Results showed that the culture filtrate of T4 increased the germination rate of millet seeds, especially during the early cultivated period. Compared with control, the undiluted, 10 and 100 times diluted culture filtrates of T4 increased the germination rates of foxtail millet seeds after 24 h-cultivation by 5.8, 6.7 and 9.2 percentage points, respectively. After being treated by the 100-times diluted culture filtrate of T4, the germination potential, germination index and radicle length of foxtail millet were increased by 9.2 percentage points, 23.1% and 13.4%, respectively. In the pot experiment, compared with control, the shoot biomass and root biomass of foxtail millet in the T4 seed coating treatment were increased by 22.5% and 32.7%, and the numbers of root tips and forks were increased by 90.9% and 66.9%, respectively. For the rhizosphere microorganisms in the seed coating with T4 treatment, the number of bacteria (B) and all microorganisms were 95.1% and 49.5% higher, with the number of fungi (F) was 52.1% less, with the ratios of actinobacteria number to fungi number (A/F), and bacteria number to fungi number (B/F) were 98.8% and 305.2% higher, respectively, whereas the ratio of actinobacteria number to bacteria number (A/B) was 52.6% less when compared with the non-inoculum adding control. The correlation analysis showed that the shoot biomass and root biomass of foxtail millet were positively correlated with the numbers of bacteria and all microorganisms, A/F and B/F in the rhizosphere soil (R=0.711~0.871, P<0.05 or 0.01), whereas negatively correlated with the number of fungi (R=-0.716~-0.738, P<0.05). The numbers of the root tips and forks were also positively correlated with B/F (R=0.690~0.882, P<0.05 or 0.01), but negatively correlated with A/B (R=-0.748~-0.810, P<0.05 or 0.01) in the rhizosphere soil. In summary, T4 promotes the growth of millet plant at jointing stage by stimulating the early germination of foxtail millet seeds, promoting the root development after germination and optimizing the microbial structure in the rhizosphere soil. This study may provide a new idea and valuable strain for the high-yield cultivation of foxtail millet.

  • HU Caixia, LIU Hong'en, NIE Zhaojun, LI Chang, QIN Shiyu, LIU Haiyang, XU Jiayang, ZHAO Peng
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 379-388. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0379
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    This paper investigated effect of combined application of phosphorus (P) with selenium (Se) on the availability and chemical fractions of Se in root-soil interface of winter wheat, which helped to investigate a possible mechanism for increase of Se uptake by winter wheat through combined application of P with Se, and to provide a theoretical basis through reasonable P and Se application in agricultural production. A rhizobox culture experiment with three P (NaH2PO4·2H2O) application rates 0 (P0), 80 (P80), 160 mg·kg-1 (P160) and two Se (Na2SeO3·5H2O) application rates 0 (Se0), 1 mg·kg-1 (Se1) was conducted to analyze the P and Se concentrations, accumulation, transport coefficient and Se chemical fractions in rhizosphere and non-rhizosphere soil of winter wheat. Results showed that, regardless of Se application, biomass and P concentration in shoots and roots of winter wheat were increased with the increasing P application. Se application (Se1) significantly decreased dry matter weight, P concentration in roots and P accumulation in each tissue. At Se1, P application (P80 and P160) significantly increased Se accumulation in each tissue, but significantly reduced Se concentration in shoots and Se migration coefficient form roots to shoots. At Se0, P160 treatment increased the concentration of exchangeable Se in rhizosphere and non-rhizosphere soil. At Se1, the concentration of exchangeable Se in the rhizosphere soil was higher than in non-rhizosphere soil, but the concentration of iron-manganese and residual Se were lower than that in non-rhizosphere soil. Appropriate combined application of P with Se promoted transformation of chemical forms of Se in soil, enhanced Se absorption and accumulation of winter wheat, which may be due to the combined effect of P application and root activity promoting transformation of Se from iron-manganese and organic sulfide bound Se to exchangeable Se to improve the bio-availability of Se in soil.

  • CHEN Hao, LI Yubo, SHAO Zhiyong, ZHENG Jirong, HU Songshen, WANG Tonglin, LIU Lihong, WANG Qiaomei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 389-396. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0389
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    To explore the effects of chitosan oligosaccharide (COS) and brassinosteroid (BR) treatment on the yield, sensory characteristics, nutritional attributes and flavor quality of tomato fruits, tomato plants at flowering and fruiting stage were foliar sprayed with four solution, i.e., control (CK), 0.5 g·L-1 COS, 0.5 g·L-1 BR, and 0.5 g·L-1 COS + 0.5 g·L-1 BR. The results showed that the combined treatment with COS and BR not only increased tomato fruit yield, but also promoted tomato coloring and softening. Compared with CK, combined treatment with COS and BR synergistically enhanced the content of lycopene, β-carotene, and vitamin C by 88.3%, 76.3%, and 28.6%, respectively. In addition, increased levels of soluble sugars (fructose and glucose) and decreased contents of organic acids (malic acid and citric acid) were observed in tomato fruits upon on COS+BR treatment, leading to higher sugar/acid ratio. The results provide a theoretical basis for improving the yield and quality attributes of tomato fruits under facility cultivation.

  • JIA Ting, CHEN Mengyun, ZHANG Lu, YI Min, GUO Shengmao, CHENG Zishan, LI Xiang, ZHONG Qiuwei
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 397-404. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0397
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    In order to clarify the truth of the nutrient distribution and stoichiometric homeostasis of organs and soil in the Pinus elliottii plantation ecosystem in Jiangxi Province, the sapling forest, young forest and mature forest in Shixi Forest Farm, Taihe County were selected as research objects. The needles, branches, roots and soil of these objects were used to explore the content of carbon (C), nitrogen (N), phosphorus (P), and ecological stoichiometry characteristics, and their stoichiometric homeostasis traits were analyzed. The results showed that the average of C, N and P contents in different organs of slash pine showed a “V” shape change with the increasing of forest age, the lowest were in the young forest which were 518.10, 3.22 and 0.71 g·kg-1, respectively. The ratio of C∶N and C∶P first increased and then decreased with the forest age, when in the young forest stage the ratios reached the peak values, which indicted that the young forest stage was the fast-growing period of slash pine, and the consumption of N and P elements was large. Soil N content and N∶P ratio increased with extension of the forest age, while soil C∶N ratio continued decreasing, indicating that the mineralization ability of soil N gradually increased with the forest age. The C∶P ratio and N∶P ratio of soil in mature forest stage were both higher than other forest ages, which meant that soil P in mature forest stage was relatively scarce. The homeostasis results of N, C∶N ratio, N∶P ratio in needles, C, C∶N ratio, N∶P ratio in branches and C, P, C∶P ratio in roots were not significant (P>0.1), and had absolute stability. The homeostasis grade of P in needles was weakly sensitive, and that in branches was sensitive. The internal stability index of N∶P (HN∶P) among different organs followed the order of roots<branches<needles, so roots were more sensitive to limiting elements in soil could be indicated. In short, during the cultivation process of slash pine plantation, the application of N, P fertilizer should be reasonable to meet the needs of rapid growth of young forest, and P fertilizers should be applied to alleviate P restriction of mature forest. The results of this study provide a theoretical reference for promoting the efficient cultivation of Pinus elliottii plantation.

  • LIU Hanghang, CHONG Peifang, MA Zhiqiang, TAN Bingbing, MA Shuai
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 405-413. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0405
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    H2S (hydrogen sulfide), as novel gas signaling molecule, plays an important role in regulating plant growth and resisting stress. In order to investigate the response mechanism of exogenous H2S to nitrogen metabolism and the optimal foliar spraying concentration of Reaumuria soongorica under salt stress, we choose the current-year R. soongorica seedlings as test material to measure the activities of nitrate reductase (NR), glutamine synthase (GS), glutamate synthase (GOGAT) and concentrations of nitrate nitrogen, soluble protein and free amino acid in leaves and roots by different concentrations of H2S (0, 0.010, 0.025, 0.050, 0.100, 0.250, 0.500 and 1 mmol·L-1 sodium hydrosulfide) under 300 mmol·L-1 NaCl stress in this pot experiment. The results showed that the concentrations of soluble protein, free amino acid and nitrate nitrogen and the activities of NR, GOGAT and GS decreased significantly compared with CK under salt stress in roots and leaves of R. soongorica seedlings. The content of soluble protein in roots and leaves of R. soongorica treated with different concentrations of exogenous H2S significantly decreased compared with the salt stress control alone (CK300), while activities of NR, GOGAT and GS, content of nitrate nitrogen and free amino acid increased significantly. Comprehensive analysis showed that NR and GS could be used as the main evaluation indexes for the alleviating effect of H2S under salt stress on R. soongorica seedlings. And the optimal alleviating concentration of exogenous H2S was 0.025 mmol·L-1 under salt stress for R. soongorica seedlings. In conclusion, the appropriate exogenous NaHS could effectively alleviate the inhibition function of nitrogen metabolism under salt stress on R. soongorica seedlings. This study provided an important theoretical basis for further understanding the mechanism of salt tolerance and breeding of high quality germplasm resources of R. soongorica.

  • WANG Hui, ZHONG Lei, FU Xiaoquan, SHI Xiang, HU Lifang, CAI Yicong, HE Haohua, CHEN Xiaorong
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 414-423. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0414
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    Early rice, especially direct seeding early rice, in the double cropping rice region of South China is vulnerable to low temperature and rainy weather such as “late spring coldness”. After stress, rice often stops growing or recovers slowly, which is easy to aggravate grass damage. In order to investigate the recovery performance of early rice after low-temperature and low-light stress and to select materials with strong rebound, 131 early rice materials were selected for this study, and the growth dynamics of seedlings were tracked for 12 d after treatment by simulating mild late spring coldness (average daily temperature of 12 ℃, 50% light for 4 d) in an artificial climate chamber. The growth recovery ability of each material was comprehensively evaluated by using principal component analysis and affiliation function method, and the representative materials were selected for the simulation test of moderate late spring coldness (average daily temperature of 10 ℃, 50% light for 4 d) to further verify the recovery performance. The results showed that all materials were classified according to growth recovery ability into strong recovery type (B556, B562, B592, etc., total 56, D value 0.549~0.699), medium recovery type (NC39, NC42, NC65, etc., total 49, D value 0.436~0.544), weak recovery type (NC54, B125, B795, etc., total 26, D value 0.265~0.415); the analysis of the differences in recovery indexes among the six high, medium and low D value materials showed that the difference between the height ratio and leaf age ratio of high D value materials and medium D value materials gradually decreased as the recovery time increased, while there was always a significant difference between low D value materials and high D value materials; at the same time, the lower the D value of materials, the later the fastest growth rate appeared and the lower the average growth rate. In the simulated experiment of moderate late spring coldness stress, the strong recovery materials screened for mild late spring coldness stress also showed strong growth recovery characteristics after moderate late spring coldness stress, but the growth indexes of each material were difficult to recover to the control level. This study provides new ideas and perspectives for early rice to cope with moderate and mild late spring coldness stress.

  • HU Yuanfeng, ZHANG Ming, FAN Shuying, GE Fei, ZHU Weifeng, XIAO Xufeng
    Journal of Nuclear Agricultural Sciences. 2023, 37(2): 424-431. https://doi.org/10.11869/j.issn.1000-8551.2023.02.0424
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    To investigate protective effects of puerarin (PUE) on cadmium (Cd) damage in pakchoi, different Puerarin (PUE) treatments with exogenous addition were taken, and the pakchoi were randomly divided into seven groups: control (CK), PUE30 (puerarin 30 μmol·L-1), Cd4 (Cd4 mg·L-1), Cd and PUE co-treatment groups (4+15, 4+30, 4+45, 4+60). After 30 d of treatment, the plant growth, Cd enrichment, stress-resistant physiological active substances including antioxidant enzymes (SOD, APX, CAT, POD) activities and hydrogen peroxide (H2O2), malondialdehyde (MDA), chlorophyll, Vc, soluble protein contents were observed, and the expression of antioxidant enzymes genes (Cu/Zn-SOD, POD, APX, and CAT) were measured by qRT-PCR. The results showed that the Cd enrichment, H2O2 and MDA contents in the roots of pakchoi were significantly higher in the Cd4 group compared with the CK group, while the root weight, fresh weight, antioxidant enzymes (SOD, POD, and APX) activities and chlorophyll content were significantly lower overall (P<0.05). Compared with the Cd4 group, the Cd+PUE co-treatment group reduced oxidative damage, significantly decreased H2O2 and MDA contents, and significantly increased chlorophyll, Vc, and soluble protein contents as well as four antioxidant enzymes activities (P<0.05); the appropriate concentration of PUE could alleviate the inhibition of Cd on the above-ground growth of pakchoi and reduce Cd uptake. In addition, the expression of Cu/Zn-SOD, POD, APX, and CAT genes in the Cd+PUE co-treatment group was higher than that in the Cd4 group, and the expression of Cu/Zn-SOD, POD, and CAT showed a increase followed by a decrease, while APX showed a decrease followed by an increase. In conclusion, the appropriate dose of Puerarin has a specific protective effect against oxidative damage under Cd stress in pakchoi. The results of this study provide a scientific basis for the application of gerbera in the prevention and control of Cd pollution and the improvement of vegetable yield.