Journal of Henan Agricultural Sciences

CONTENTS

2024, 53(11): 0.

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Application Status and Research Progress of Bioaugmentation Technology in Agricultural Waste Composting Treatment

TIAN Jingjie, CUI Erping, LIU Chuncheng, HU Chao, LI Zhongyang, CUI Bingjian

2024, 53(11): 1-16. DOI: 10.15933/j.cnki.1004-3268.2024.11.001

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Composting treatment is one of the crucial approaches to achieve the harmlessness and resource utilization of agricultural wastes.Nevertheless,conventional composting methods suffer from issues such as a prolonged fermentation period,low conversion efficiency,significant nutrient loss，difficult control of secondary pollution,as well as the emission of odors and greenhouse gases.Microorganisms have emerged as effective means to address the aforementioned problems due to their characteristics of promoting decomposition,deodorization,emission reduction,and enhancing fertilizer efficacy.Regarding the bioaugmentation technology for composting using agricultural wastes as raw materials，this paper summarizes the application and research progress of bioaugmentation technology in the composting process of agricultural wastes at home and abroad from aspects such as the types of functional microbial strains and screening,the construction of complex microbial agents,the functions of microorganisms,and the influencing factors and mechanisms of bioaugmentation.Moreover,it explores the existing problems in the composting process of agricultural wastes and proposes the development trends and countermeasure suggestions for the resource treatment and utilization of agricultural wastes.

Research Progress of Swine Body Mass Estimation Based on Machine Vision

WANG Xingjia, WEI Qinggang, MU Yuanjie, SHENG Qingkai, YANG Liang, AN Jing, ZHANG Xia, ZHENG Jiye

2024, 53(11): 17-26. DOI: 10.15933/j.cnki.1004-3268.2024.11.002

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In swine breeding,body mass is one of the most important index parameters,so it is of great significance to obtain timely and accurately body mass information of swines. With the rapid development of machine vision technology,the non‑contact body mass estimation of live swines has been widely concerned.At present,non‑contact body mass estimation has achieved some research results,but because of the complexity of the real breeding environment,the real‑time estimation of swine body mass in the farm still faces various challenges.This paper reviews the research progress and application of machine vision technology in swine body mass estimation in the past 20 years at home and abroad,and analyzes the existing problems and future research priorities,aiming to provide references for the intelligent transformation and upgrading of modern animal husbandry in China.

Cloning of γ‑Gliadin 1 Gene and Screening of Its Interacting Proteins in Wheat

WANG Shasha, HE Ning, HUANG Chao, WANG Gang, WANG Qingchang, CHAO Yueen

2024, 53(11): 27-33. DOI: 10.15933/j.cnki.1004-3268.2024.11.003

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In order to further study the molecular mechanism of γ‑gliadin 1 gene in regulating flour quality of wheat,the γ‑gliadin 1 gene was cloned using Zhengmai 158(low protein content and high dough strength)as materials,bait vector pGBKT7‑γ‑gliadin 1 was constructed,the interacting proteins of wheat γ‑gliadin 1 were screened from Zhengmai 158 cDNA library by the yeast two‑hybrid system,and rotary test was used to validate the interaction between key candidate proteins and γ‑gliadin 1.The results showed that the γ‑gliadin 1 gene was successfully cloned with the cDNA length of 1 020 bp.A total of 117 blue clones were screened by the yeast two‑hybrid system.Ten possible candidate interacting proteins of γ‑gliadin 1 were obtained through colony PCR detection,sequencing,and homology analysis using the BLAST on NCBI,which were cysteine‑rich and transmembrane domain‑containing protein 1‑like(XM_044546254.1),alpha‑amylase/trypsin inhibitor‑like(XM_044509330.1),lichenase 2(XM_044550285.1,XM_044594213.1),sucrose 1‑fructosyltransferase‑like(XM_044554537.1),pullulanase 1(XM_044577151.1),ervatamin‑B‑like(XM_044577994.1),cell number regulator CNR8‑like(XM_044483337.1,XM_044491902.1)and CNR2‑like(XM_044468782.1)related to plant reproductive growth(especially development of fruit or grain),and ubiquitin domain‑containing protein DSK2b‑like（dominant suppressor of KAR 2b like,XM_044546840.1)and CIP73(CCa MK‑interacting protein of 73 ku,XM_044554473.1),respectively.The rotation validation assays of candidate interacting proteins indicated that four candidate proteins were probably interacted with γ‑gliadin 1,which were cysteine‑rich and transmembrane domain‑containing protein 1‑like,alpha‑amylase/trypsin inhibitor‑like,lichenase 2 and ervatamin‑B‑like,respectively.Therefore,it is speculated that γ‑gliadin 1 may mainly interact with these candidate proteins to be involved in the synthesis and degradation of storage proteins(such as cysteine)and starch in wheat grains.

Comprehensive Evaluation of Stem Lodging Resistance of 145 Maize Inbred Lines

ZHANG Fengqi, ZHANG Jun, MA Zhiyan, WU Xiaopan, ZHOU Qingwei, MU Xinyuan, TANG Baojun, LI Xin, XIA Laikun, DING Yong

2024, 53(11): 34-40. DOI: 10.15933/j.cnki.1004-3268.2024.11.004

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The plant height(PH),ear height(EH),stem rind penetrometer resistance(RPR),internode diameter(ID)and internode length(IL)were investigated under multiple environments with 145 maize inbred lines from home and abroad as materials.The excellent inbred lines with strong stem lodging resistance were screened out through correlation analysis,principal component analysis and cluster analysis,so as to provide excellent germplasm resources for maize lodging resistance breeding.The results indicated that PH,EH,RPR,ID and IL were extremely significantly different among different maize inbred lines.The heritability of PH with 0.99 was the highest,followed by EH and RPR all with 0.97.The correlation analysis results showed that RPR was extremely significantly positively correlated with PH.The principal component analysis results showed that the first three principal components represented 77.02% of the information of the five traits of 145 maize inbred lines,and their contribution rates were 37.08%,21.94% and 18.00%,respectively. Based on the comprehensive evaluation and cluster analysis of lodging resistance of each inbred line,145 inbred lines were divided into 6 groups,13 inbred lines with strong lodging resistance,such as S268,S112,S135,S270,etc,were screened out.

Effect of Reducing Nitrogen and Increasing Density on Yield and Nitrogen Absorption and Utilization of Rice in Cold Region

JIAO Feng, ZHANG Yuhan, DONG Wenjun, TANG Ao, LIU Youhong, MENG Ying, WANG Shuo

2024, 53(11): 41-49. DOI: 10.15933/j.cnki.1004-3268.2024.11.005

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The long‑term experiment with reduced nitrogen and increased density for rice in cold region under straw return was started in 2017. A total of the following five treatments were set up，CK：conventional density（30 cm×13. 3 cm），no N fertilizer；T1：conventional density（30 cm×13.3 cm），conventional N application rate（180 kg/ha）；T2：increasing density by 32%（30 cm×10 cm），conventional N application rate（180 kg/ha）；T3：increasing density by 32%（30 cm×10 cm），20% reduction in total conventional N application rate by spike fertilizer（total N application rate of 144 kg/ha）；T4：increasing density by 32%（30 cm×10 cm），20% reduction in total conventional N application rate by basal fertilizer（total N application rate of 144 kg/ha）. The experiment data in 2023 was used to study the effect of reducing nitrogen and increasing density on yield and nitrogen absorption and utilization of rice.The results showed that T4 treatment had the highest yield，which was significantly increased by 39.6%，9.8% and 13.3% compared with CK，T1 and T2 treatments respectively；T3 treatment was the second，showing no significant difference from T4 treatment. The increase of yield of the two treatments was mainly attributed to the increase of effective spikes number，and T4 treatment significantly increased by 64.0% and 18.5% compared with CK and T1 treatment respectively.The harvest indexes of T3 and T4 treatments were higher，and the difference between them was not significant，but both were significantly higher than the other treatments，among which T4 treatment was 30.4%，53.3% and 15.6% higher than CK，T1 and T2 treatments respectively. From jointing stage to mature stage，T4 treatment generally had the highest SPAD value of leaves，the lowest attenuation rate of SPAD，and higher dry weight of above‑ground part. T1—T4 treatments significantly increased nitrogen uptake by 53.6%—94.2% compared with CK，and the increase was the greatest in T4 treatment. Compared with the T1 treatment，the nitrogen fertilizer uptake and use efficiency，nitrogen uptake rate，nitrogen fertilizer agronomic utilization rate and nitrogen partial productivity of T4 treatment significantly increased by 36.1%，30.1%，82.4% and 37.3%，respectively，whereas apparent surplus and deficit of soil nitrogen and nitrogen surplus rate significantly decreased by 87.5% and 103.8%，respectively；the nitrogen fertilizer uptake and use efficiency of T3 treatment significantly decreased by 22.5%，while nitrogen fertilizer agronomic utilization efficiency and nitrogen partial productivity significantly increased by 71.2% and 34.9%.Nitrogen partial productivity，nitrogen fertilizer agronomic utilization rate，nitrogen uptake rate were significantly positively correlated with yied，and nitrogen surplus rate and soil nitrogen apparent surplus and deficit were significantly negatively correlated with yield. In conclusion，increasing density while reducing the amount of basal nitrogen fertilizer can improve rice yield as well as nitrogen uptake and utilization rate，which is more suitable for the growth of rice in cold region.

Effects of Sowing Date and Enhancing Fragrance Fertilizer on Yield，Quality and Fragrance of Aromatic Rice with High Quality

2024, 53(11): 50-62. DOI: 10.15933/j.cnki.1004-3268.2024.11.006

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The effects of sowing dates［conventional sowing date（S1）and delayed sowing date（S2）］and enhancing fragrance fertilizer［no enhancing fragrance fertilizer（conventional fertilization，F1） and enhancing fragrance fertilizer（F2）］ on yield，quality and fragrance of major aromatic rice varieties ［Yexiangyouyousi（V1），19xiang（V2）and Huaxiaxiangsi（V3）］were studied in Hubei Province，so as to provide a scientific basis for the production of high‑quality fragrant rice. The results showed that the high temperature and heat damage could be avoided or alleviated by delaying the sowing date，and the initial heading stage was postponed to mid‑to‑late August after delaying the sowing date.The processing quality and appearance quality of aromatic rice improved，and the content of aromatic substance 2‑AP in brown rice significantly increased by 12.14%—162.84% after delaying the sowing date.The processing quality of aromatic rice could be improved by adding enhancing fragrance fertilizer，but the appearance quality was slightly affected.Meanwhile，the content of aroma substance 2‑AP in brown rice could be increased by 6.19%—40.60%.Daily maximum temperature was negatively correlated with chalky grain rate and chalkiness，total rainfall was extremely positively correlated with yield and 2‑AP content in brown rice，and daily maximum temperature，daily minimum temperature and days of daily maximum temperature≥35 ℃ were extremely negatively correlated with 2‑AP content in brown rice. Overall，delayed sowing date and enhancing fragrance fertilizer are beneficial to improving the processing quality and enhancing fragrance，but the enhancing fragrance fertilizer needs to be properly combined according to the characteristics of aromatic rice varieties and sowing date to achieve better results.

Identification and Expression Analysis of PAL Gene Family in Tartary Buckwheat

WANG Qinghai, JIA Yanhua, LUO Yanran, WANG Anhu

2024, 53(11): 63-73. DOI: 10.15933/j.cnki.1004-3268.2024.11.007

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Based on bioinformatics methods，the PAL gene family in tartary buckwheat was identified and analyzed at the whole genome level. The gene expression levels of PAL family in different tissues were detected using quantitative real‑time PCR（qRT‑PCR），so as to provide a reference for further investigation into the fundamental characteristics and gene functions of this gene family.The results showed that the PAL gene family in tartary buckwheat consisted of five members distributed across four chromosomes，with amino acids of encoding proteins ranged from 688 to 722 aa. All proteins exhibited
similar physicochemical properties，including molecular weights ranging from 74 453.74 u to 78 377.58 u and isoelectric points ranging from 5.64 to 6.28.Furthermore，PAL members were stable and hydrophilic proteins，and were located in the cytoplasm. Collinearity analysis showed that there were six collinear gene pairs between tartary buckwheat PAL genes and soybean PAL genes，and three collinear gene pairs between tartary buckwheat PAL genes and alfalfa PAL genes. PAL gene family shared high protein sequence homology，with strictly conserved secondary structures and motifs. All PAL proteins contained a MIO（4‑methylidene‑imidazole‑5‑one）active group composed of Ala‑Ser‑Gly.PAL proteins from different species were relatively independent and conserved during evolution. PAL gene family member possessed many cis‑acting elements regulating light response，hormone response，stress response and development in promoter.Expression pattern analysis results showed that five PAL genes were expressed in different degrees in tartary buckwheat tissues.Notably，the FtPAL2 gene showed specific and highly elevated expression levels in flower organs.

Bioinformatics and Abiotic Stress Response of Tobacco NtTCHQD1

XIE Min, ZHANG Jingyao, WU Guizhi, CHEN Gang, YANG Shuxian, TANG Qin, LIU Renxiang

2024, 53(11): 74-82. DOI: 10.15933/j.cnki.1004-3268.2024.11.008

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Glutathione S‑transferase（GST）plays a crucial role in plant resistance to abiotic stress.To investigate the potential biological function of the NtTCHQD1 gene in the tetrachlorohydroquinone dehalogenase（TCHQD）subfamily in tobacco，with the aim of providing a novel gene resource for the development of tobacco resistance germplasm and variety selection，the biological information of NtTCHQD1 was analyzed，and the tobacco variety K326 was used as material to be treated by nine kinds of stress，including water，temperature and hormone.The response characteristics of NtTCHQD1 gene in different treatments were analyzed by measuring the expression levels of NtTCHQD1 in different treatments.The results indicated that the NtTCHQD1 protein was an unstable，non‑secretory，and hydrophilic protein with an instability coefficient of 40. 52 and no signal peptide.The NtTCHQD1 protein mainly comprised serine，threonine，and tyrosine phosphorylation sites，which might be phosphorylated by these protein kinases，thereby participating in various physiological and biochemical processes of tobacco.The analysis of promoter cis‑acting elements revealed that NtTCHQD1 contained hormone，anaerobic，drought，light，and other response elements，which might be involved in plant hormone signal transduction，light and temperature regulation，plant growth and development，and abiotic stress responses.The expression of NtTCHQD1 was elicited under diverse stress treatments，particularly in response to drought，high temperature，and low temperature.The responses of tobacco plants to various stresses exhibited tissue specificity，among which the response of roots to salicylic acid（SA）and gibberellin（GA₃）was the most susceptible，and the response of leaves to other stress treatments except for SA and GA₃ was the most vigorous. In conclusion，NtTCHQD1 might regulate the response of tobacco to hormone and abiotic stress during its growth and development，and could be employed as a novel gene resource for tobacco and other crops to breed outstanding varieties resistant to drought and extreme
temperature.

Effect of Continuous Application of Municipal Standardized Sewage Sludge on the Risk of Heavy Metal Pollution in Apples and Orchard Soil

BAI Jiayun, XU Junxiang, SUN Qinping, YAO Hai, YANG Yingjie, HE Liang, GAO Shitao, XIONG Jianjun, LIU Yanpeng

2024, 53(11): 83-92. DOI: 10.15933/j.cnki.1004-3268.2024.11.009

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To investigate the impact of continuous application of municipal sludge on the risk of heavy metal contamination in apples and orchard soil，a field experiment was conducted with four treatments：conventional fertilization（T1），moderate sludge application（T2），high sludge application（T3）and low sludge application（T4）.The study analyzed the risk of heavy metal contamination in apples and soil under different sludge application rates and durations.The results indicated that the concentrations of Cd，Cr，Pb，Hg，and As in apples did not exceed the regulatory limits.After three years of sludge application，the single‑factor pollution indexes for heavy metals in the soil were below the warning thresholds by 71.98% to 75.98%，66.57% to 69.90%，92.32% to 94.01%，78.59% to 90.40%，and 17.22% to 23.29%，respectively.The Nemerow comprehensive pollution index and potential ecological risk index were also below the warming thresholds by 9.51% to 16.08% and 82.50% to 86.75%，respectively.Following three years of sludge application，the single‑factor pollution index for Hg and As increased by 29.52 times and 83. 14% compared to background levels. The potential ecological risk index for treatments T2 to T4 increased by 78.38%，95.69% and 48.18% respectively，compared to background values. The potential ecological risk contribution rate of Hg increased from 11.00%—17.02% to 19.32%—33.70%，while As increased from 21.55%—24.61% to 31.54%—39.17%. Based on the calculations for Cd and Hg，the safe application duration for sludge was relatively shorter，ranging from 32.30 to 116.31 years and from 36.07 to 129.91 years，respectively. In summary，after the continuous application of sewage sludge for three years，both apple fruit and soil remained in an uncontaminated state. However，with the increasing of sludge application duration，the single‑factor pollution index for Hg and As，comprehensive pollution index and potential ecological risks all increased，with Hg and As being the primary contributing factors.Based on the safe application duration of sludge，Cd and Hg emerged as the most significant limiting elements.The rational utilization of sludge does not lead to heavy metal contamination in the soil.

Diversity of Arthropods and Population Dynamics of Dominant Pests on Polygonatum kiugiauum under Photovoltaic Panels

SUN Ganlin, CAI Haosheng, XIAO Guanli, HE Fengchun, HE Yanyan, YANG Huquan, YU Zhigang, CHEN Bin

2024, 53(11): 93-100. DOI: 10.15933/j.cnki.1004-3268.2024.11.010

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To clarify the structure and diversity of the arthropod community on Polygonatum kiugiauum planted under photovoltaic panels，and provide technical support for the integrated pest management on P.kiugiauum under photovoltaic panels and the development of photovoltaic agriculture，a systematic investigation on the species was conducted using the visual survey and blue sticky card trapping in P.kiugiauum planting areas under photovoltaic panels and open‑field planting in 2023，the community diversity of arthropods was evaluated using the community diversity index and population dynamics of major pests was analyzed.The results showed that there were 27 species of arthropods belonged to 27 genera，25 families，10 orders and 2 classes on P.kiugiauum under the photovoltaic panels.Additionally，there were 30 species of arthropods in 30 genera，28 families，10 orders and 2 classes under open‑field planting of P.kiugiauum.During the whole investigation period，the diversity index and evenness index of arthropod community on P.kiugiauum planted under photovoltaic panels were higher than those planted in the open‑field and followed a dynamics of decrease‑increase，and the dominance index followed an increase‑decrease dynamics.The richness index of arthropod community on P. kiugiauum under photovoltaic planting followed a dynamics of increase‑decrease‑increase，while open‑field planting followed a dynamics of increase‑decrease‑constancy.The similarity of arthropod communities on P.kiugiauum was high with a value of 0.84 under the two planting modes.The dominant species of arthropods was Tetranychus kanzawai，whose population showed a single‑peak pattern. However，the population under the photovoltaic panels was significantly lower than that in open‑field planting，with a peak of 6.77 in May under the photovoltaic panels，and a peak of 14.75 in July under open‑field planting，respectively.In conclusion，the diversity of arthropod communities on P.kiugiauum planted under photovoltaic panels was higher than that under open‑field planting，and it was favourable for the control of the dominant pest T.kanzawai.

Screening and Safety Evaluation of Post‑emergence Herbicides for Controlling Broadleaf Weeds in Sesame Fields

SU Wangcang, NIU Yujia, YAN Zhaoling, DU Li, XUE Fei, SUN Lanlan, XU Hongle, WU Renhai, LIU Hongyan

2024, 53(11): 101-108. DOI: 10.15933/j.cnki.1004-3268.2024.11.011

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To explore safe and efficient chemical weed control technologies for sesame after emergence，the herbicides that exhibit high safety for sesame and have good control effects on broadleaf weeds were screened through indoor potting experiments and field trials. The results of the indoor trials showed that among the 17 broadleaf herbicides，only isoproturon and bentazone were discovered to be highly safe for sesame.Futher，the safety of isoproturon and bentazone to sesame and their control efficacy against broadleaf weeds were evaluated through field trials. The results showed that the treatment of isoproturon at 420—630 g/ha（active ingredient，the same below）caused phytotoxicity in sesame，such as the emergence of drug spots and growth inhibition，etc. Nevertheless，sesame growth recovered to normal within 30 d after the treatment. Sesame treated with 336—420 g/ha bentazone had significantly higher plant height than the control treatment at 30 d after the treatment，but was significantly lower than the artificial weeding treatment. The total plant control efficacy of isoproturon at 420—630 g/ha against broadleaf weeds such as Acalypha australis L.and Clinopodium polycephalum Vaniot was 84.00%—90.40%，the total fresh weight control efficacy was 87.62%—94.30%，and the sesame seed yield increased by more than 37%.The control efficacy of bentazone at a dosage of 336—504 g/ha against Acalypha australis L.was unsatisfactory，with the total plant control efficiency being lower than 40% and the total fresh weight control efficiency being lower than 57%，and sesame yield was merely increased by 11.53%—14.72%.In conclusion，the application of isoproturon at 420—630 g/ha during the 5—6 leaves stage of sesame is safe and can be used for the control of broadleaf weeds in sesame fields，presenting significant application prospects.

Distinguishment of the Instars of Macrosiphoniella yomogicola in Artemisia argyi

XU Lanjie, AN Sufang, YU Yongliang, YANG Qing, TAN Zhengwei, LI Chunming, SU Xiaoyu, SUN Yao, LIANG Huizhen

2024, 53(11): 109-116. DOI: 10.15933/j.cnki.1004-3268.2024.11.012

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The morphologic indicators of body length，body width，head width，antenna length，cornicle length，cauda length，and hind tibia length for Macrosiphoniella yomogicola at 1st，2nd，3rd，4th instars and adult were measured by stereoscopy，the data were analyzed by correlation analysis and regression analysis，and then the characteristics of Macrosiphoniella yomogicola at different instars were identified，to lay technical support for aphid integrated control in Artemisia argyi of Chinese medicinal materials.The results showed that morphologic indicators of Macrosiphoniella yomogicola had significant differences at different instars，and showed significantly positive correlation among them. Regression analysis indicated that head width，antenna length，cornicle length and hind tibia length of Macrosiphoniella yomogicola were linearly positively related to instars，but body length，body width and cauda length showed no linear significant correlation with instar. Further data analysis showed that cornicle length of wingless and winged Macrosiphoniella yomogicola displayed no overlap between adjacent instars，hind tibia length displayed no overlap between 1st and 2nd instar nymphs，antenna length displayed no overlap between 1st，2nd and 3rd instar nymphs，but body length，body width，head width and cauda length had overlap between adjacent instars. In conclusion，cornicle length can be screened as a main indicator for identification of instars of Macrosiphoniella yomogicola，and antennae length and hind tibia length can be as assistant indexes.

Identification and Expression Analysis of Auxin Response Factor（ARF）Gene Family in Tree Peony

ZHAO Yinge, YUAN Xin, GAO Jie, WANG Huijuan, LI Yanmin, WANG Xiaohui, SHI Man, FENG Naixi, FU Zhenzhu, JIA Wenqing

2024, 53(11): 117-126. DOI: 10.15933/j.cnki.1004-3268.2024.11.013

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To study the auxin response factor（ARF）gene family in tree peony，the gene family was identified and bioinformatics analysis was conducted.The results showed that a total of 19 ARF genes were identified in P.sufruticosa‘Shima Nishiki’，P.ostii and P.sufruticosa‘Luoshen Xiaochun’，and the ARF gene family members of tree peony were divided into four subfamilies based on the evolutionary classification of Arabidopsis thaliana ARF gene family，and according to the phylogenetic tree，there was a close evolutionary relationship between peony and Arabidopsis thaliana ARF homologous genes.There were differences in the length and isoelectric point of ARF proteins among different tree peony cultivars.All PsARFs contained conserved B3 and Auxin_resp domains，and most PsARFs also contained Aux/IAA domains.Conserved motif analysis showed that the number of Motif was 4—12，and gene structure analysis showed that the number of introns in the ARF gene of peony was 1—19，and there were some differences in the number of Motif and introns among different subfamilies.Tissue expression characteristics analysis showed that all PsARF genes were expressed in roots，stems and leaves of peony tissue culture seedlings，except that PsARF2b was not expressed in various tissues，but the expression levels in different tissues were very different.The expression levels of PsARF7，PsARF19，PsARF8a，PsARF8b，PsARF6，PsARF11a，PsARF11b，PsARF9，PsARF16b，PsARF17，PsARFx were the highest in roots，PsARF5，PsARF3 and PsARF4 were the highest in stems，and PsARF2a，PsARF1，PsARF16a and PsARF10 were the highest in leaves.

Effect of Selfing on Soluble Sugar Content of Chestnut

MA Yate, YUAN Guomei, LI Zhe, LI Huiying, YU Liyang, HUANG Ruimin, BAO Minhu, DU Hongru, WANG Dongsheng

2024, 53(11): 127-134. DOI: 10.15933/j.cnki.1004-3268.2024.11.014

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This study explored the effect of self‑pollination and natural pollination on the content of soluble sugar components of chestnut in Yanshan area，and excavated the information of phenotypic traits，to provide theoretical support for the breeding of high‑sugar varieties of chestnut. Yanshan Zaofeng，Yanlu，Tafeng，Yanzi and Beijing 8 were self‑pollinated and naturally pollinated，respectively.The contents of sucrose，maltose，inositol，glucose，D‑galactose，D‑fructose and raffinose were determined and the biochemical sweetness was calculated.Then the difference analysis，variation analysis，correlation analysis，OPLS‑DA analysis and cluster analysis were carried out.The results showed that the content of soluble sugar components in most chestnuts was significantly different between self‑pollination and natural pollination treatments. The inositol content of chestnut was significantly increased after selfing.The content of sucrose was the highest and the content of D‑galactose was the lowest in the soluble sugar components of chestnut.The variation coefficient of raffinose was the highest，which was 72.197 7% in self‑pollination and 68. 826 8% in natural pollination.After selfing，the coefficient of variation of D‑fructose decreased，and the coefficient of variation of other soluble sugar components increased.Sucrose was significantly positively correlated with biochemical sweetness，and D‑galactose was significantly positively correlated with raffinose. After chestnut selfing，glucose and maltose also showed a very significant positive correlation.The biochemical sweetness of chestnut increased after selfing.Four differential soluble sugar components were screened by OPLS‑DA analysis：sucrose，raffinose，glucose and inositol.Cluster analysis showed that the content of differential soluble sugar components in chestnut after selfing was generally higher than that of natural pollination. The variation of soluble sugar components in chestnut after selfing is rich，and the soluble sugar components of different chestnut resources are quite different，which provides theoretical support for the optimization of germplasm resources and the improvement of chestnut sugar content.

Development of Indirect Enzyme‑Linked Immunosorbent Assay Based on the Dimeric Nucleocapsid Protein for Detecting Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

ZHAO Xiao, QIAO Songlin, SUN Yaning, QIAN Qisheng, LI Rui, ZHANG Gaiping

2024, 53(11): 135-146. DOI: 10.15933/j.cnki.1004-3268.2024.11.015

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In order to prepare N protein with good stability and immunoreactivity，N‑dimer was designed and expressed，and an iELISA detection method for PRRSV antibody was established based on this protein.Firstly，the NADC30‑like N protein gene sequence of the current dominant PRRSV strain in China was selected，and the N protein gene sequence was concatenated through a（GGGGS）3 linker.After codon optimization and synthesis，an N‑dimer recombinant expression vector was constructed.The recombinant protein was expressed using prokaryotic expression system and purified by nickel ion chelation affinity chromatography.Its reactivity with PRRSV positive and negative sera was analyzed by Western‑blot.Its stability after freezing and thawing and immunoreactivity were further compared with those of N‑monomer.An iELISA was subsequently developed based on N‑dimer as coating antigen，and evaluated for the specificity，sensitivity，and repeatability.The developed iELISA was finally applied to clinical serum sample detection.The results showed that under the conditions of 16 ℃ and 0.2 mmol/L isopropylthiogalactoside，N‑dimer was obtained with a large amount of soluble expression after 16 hours of induction.Western‑blot results showed that N‑dimer could specifically react with PRRSV positive serum；Compared to N‑monomer，N‑dimer coule be stably stored and had better immunoreactivity.We successfully established a PRRSV antibody iELISA detection method based on N‑dimer，which had good specificity，sensitivity，and repeatability；Compared with the results of commercial test kits，the positive agreement rate was 96.88%，and the overall agreement rate was 96.86%，indicating a high agreement rate.In summary，this study successfully prepared N‑dimer with good stability and excellent immunoreactivity；A PRRSV antibody iELISA detection method was established based on N‑dimer.

Expression Analysis of gga‑miR‑103‑3p in Different Tissues and Cells in Chicken and Screening of Its Key Target Genes

HUANG Huayun, LI Ruirui, YANG Miaomiao, ZHAO Zhenhua, WANG Qianbao, LIANG Zhong, HUANG Zhengyang, WU Zhaolin, LI Chunmiao, HAN Wei

2024, 53(11): 147-155. DOI: 10.15933/j.cnki.1004-3268.2024.11.016

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In order to preliminarily elucidate the regulatory role of gga‑miR‑103‑3p in fat deposition and muscle development in chickens，fluorescence quantitative PCR（qPCR）was used to detect the expression of gga‑miR‑103‑3p in abdominal fat，liver，leg muscle，abdominal adipocytes，intramuscular adipocytes and adult myoblasts in chickens of the S3 lineage and recessive white‑feathered（RR）chickens，and its key target genes were screened.The results showed that gga‑miR‑103‑3p was expressed in liver，abdominal fat，leg muscle，abdominal adipocytes，intramuscular adipocytes and adult myoblasts，and there were differences among tissues and breed（line）.In the liver，gga‑miR‑103‑3p expression level was significantly higher in S3 line chickens than that in RR chickens at 0 week of age（0W）.For S3 chickens，gga‑miR‑103‑3p expression level was the highest at 16 weeks of age（16W），which was significantly higher than those at 0W，2 weeks of age（2W），8 weeks of age（8W）and 14 weeks of age（14W）；For RR chickens，gga‑miR‑103‑3p expression level was significantly higher at 16W and 14W than those at 0W and 8W. In abdominal fat，gga‑miR‑103‑3p expression level was significantly higher in RR chickens than that in S3 chickens at 0W. gga‑miR‑103‑3p expression level was significantly higher at 0W than those at 2W，8W，14W and 16W in S3 and RR chickens，and decreased in S3 chickens with the advancement of developmental period.In the leg muscle，the expression level of gga‑miR‑103‑3p was significantly higher in RR chickens than that in S3 chickens at 16W，and vice versa at 0W；The expression level of gga‑miR‑103‑3p in S3 chickens generally declined with the advancement of developmental period，and the expression level was significantly higher at 0W than those at other weeks of age；The expression level of gga‑miR‑103‑3p in RR chickens first declined and then increased with the advancement of developmental period，and the expression level was significantly higher at 16W than those at other weeks of age. In abdominal adipocytes and intramuscular adipocytes，the expression levels of gga‑miR‑103‑3p at 4 d of differentiation and 6 d of differentiation were significantly higher than that at the proliferative stage；In adult myoblasts，the expression level of gga‑miR‑103‑3p increased with the extension of differentiation，and the expression levels of gga‑miR‑103‑3p were significantly higher at 5 d of differentiation and 7 d of differentiation than that at the proliferative stage.The GO，KEGG enrichment and protein interaction analyses of gga‑miR‑103‑3p target genes showed that FBXW7，CDK6，NF1 and PIK3R1 were important target genes of gga‑miR‑103‑3p，of which FBXW7 and PIK3R1 were particularly critical.In conclusion，gga‑miR‑103‑3p may affect muscle development and fat deposition through FBXW7，CDK6，NF1 and PIK3R1，of which FBXW7 and PIK3R1 are particularly critical.

Quantitative Detection Method of Grain Storage Height in Grain Silos Based on U‑Net

REN Feiyan, ZHANG Hui, LI Zhi, YANG Weidong, ZHANG Yanfei, CHEN Weidong, TAN Yunjian, LIU Ruiyun

2024, 53(11): 156-163. DOI: 10.15933/j.cnki.1004-3268.2024.11.017

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Due to the distribution of grain storage in China is characterized by many points，long lines and wide areas，the traditional method of monitoring grain quantity is struggling with low efficiency，high costs，and significant delays.There is an urgent need to research and develop a new technology that can perform real‑time and rapid detection of the quantity of grain stored in grain silos.To address the problem，we proposed a quantitative detection method of grain storage height in grain silos based on U‑Net.By segmenting the grain surface and ventilation windows in pictures captured by monitoring cameras in the grain warehouse，we obtained pixel values of the edge of the grain surface and the upper and lower edges of the ventilation window.Taking into account the height of the ventilation window’s upper and lower edges above and below the ground，we could determine the actual height of grain storage in the grain silo. Subsequently，merged with the length，width，grain density，and other fundamental data，we obtained the actual quantity of grain stored in the grain silo.In this study，the height of grain storage was calculated by analyzing and processing the segmented soil images using U‑Net，DeepLabV3+，and PSPNet algorithms respectively.The experimental results showed that the mean intersection over union（MIoU）reached 93.25%，which was 1.82 and 2.69 percentage points higher than that of DeepLabV3+and PSPNet，respectively.The mean pixel accuracy（MPA）reached 95.88%，which was 2.42 percentage points higher compared with PSPNet.The quantitative analysis error of U‑Net was 3.51%，which was 1.34 and 0.43 percentage points lower compared with DeepLabV3+ and PSPNet，respectively.U‑Net was more suitable as a segmentation algorithm for quantitative calculation of grain storage height in granaries.It is not necessary to set up the measuring scale beforehand and with this method.By relying solely on the monitoring camera inside the warehouse，the height measurement of grain pile can be achieved，which can effectively meet the fast detection requirements for the quantity of grain stored in the warehouse.

Effects of Different Harvesting Periods on the Quality of Suli Pear and Fresh‑keeping Effect in Semi‑underground Ventilation Storeroom

WU Zhongying, ZHANG Sipu, CUI Wei, GUO Chaofeng, LU Yunfeng, XIE Yuanming, NIU Jiajia

2024, 53(11): 164-172. DOI: 10.15933/j.cnki.1004-3268.2024.11.018

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In order to determine the suitable harvesting date of Suli pear，Jindingxiehua Suli pear from Ningling County was taken as the raw material. The storage was carried out in semi‑underground ventilation storeroom after harvest in five periods（at 140，152，166，180，and 190 days after full flowering，respectively numbered 1—5）.Color，hardness，marketable fruit rate，total soluble solid（TSS）content，titratable acid（TA）content，malondialdehyde（MDA） content and polyphenol oxidase（PPO） activity were test during storage. Meanwhile，temperature and humidity in the ventilation storeroom during storage were monitored.The storage property was evaluated by membership function method.The results showed that with the delay of harvest date，fruit hardness decreased，TSS content increased first and then decreased，TA content decreased，and single fruit weight，L^*，a^*and b^* values increased. When stored for 130 days，the good fruit rate of the other harvesting periods was above 80% except for harvesting period 2.At 160 days of storage，harvest period 3 had best storage performance，with the highest values of marketable fruit rate（80.6%），L^*（79.00）and TSS content（11.53%）.The PPO activity［21.4 U（/ min·g）］was the lowest.The a* value was 7.38，the b^* value was 47.68，the hardness was 4.94 kg/cm²，the TA content was 0.056%，and the MDA content was 12.81 mmol/g.The average membership value of the indicators related to storage property of the five periods was calculated，and then their storage property was ranked as harvest period 3>harvest period 2>harvest period 1>harvest period 4>harvest period 5.Therefore，it is recommended that for Ningling Jindingxiehua Suli pear with semi‑underground ventilation storeroom storage，the suitable harvest date is September 15，166 d after the flowering stage，and the suitable storage period is mid‑February of the next year，with early March of the next year to complete sales.

Effect of Mechanical Properties of Flue‑cured Tobacco on the Quality of Threshing and Redrying Lamina

ZHOU Lijun, WANG Xun, ZHU Bo, GUAN Xin, HU Zongyu, GAO Yiping, LI Huan, ZHAO Sensen, ZHANG Yuhai

2024, 53(11): 173-180. DOI: 10.15933/j.cnki.1004-3268.2024.11.019

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In order to investigate the effect of mechanical properties of flue‑cured tobacco on the quality of threshing and redrying processing，C3F grade tobacco leaves produced from three production areas，Chenzhou in Hunan Province，Bijie in Guizhou Province and Sanming in Fujian Province were selected to analyze their mechanical properties，the area distribution and shape characteristics of the tobacco leaves after threshing and redrying processing were tested，and the relationships between the mechanical properties and the quality indicators of the tobacco leaves were analyzed using relevant analysis.The results showed that other mechanical indicators of tobacco leaves from three different production areas showed significant differences，except for tensile strength，elongation，and stem content.Tobacco leaves in Chenzhou had higher shear strength and penetration strength，tobacco leaves in Bijie had the highest adhesion force，while tobacco leaves in Sanming had higher leaf stem binding force and branch binding force.The overall area of tobacco leaves in Chenzhou was relatively larger，with a high proportion of area over 500 mm².Tobacco leaves in Bijie had good uniformity，with a high proportion of tobacco with an area of 300—500 mm².The area of tobacco leaves in Sanming was relatively smaller，the proportion of tobacco with an area of less than 300 mm2 was significantly higher than other areas.There were no significant differences in rectangularity，thinness，and circularity. The proportion of lamina area distribution in different production areas showed a trend of first increasing and then decreasing with the increase of lamina area，and the difference in shape characteristic indicators（Xc）was not significant，with slight differences in the proportion peak value. Among them，tobacco in Bijie had the highest proportion peak value with a rectangular degree of 0.63 and a contour temperature characteristic value of 3.75，while tobacco in Sanming had the highest proportion peak value with a circular degree characteristic value of 2.79.There was no significant difference in the proportion peak value of ellipticity characteristic values among different areas of tobacco.The characteristic area of tobacco was significantly positively correlated with shear strength，was weakly correlated with tensile strength，adhesion，stem ratio，and strongly correlated with other indicators；The uniformity coefficient was significantly negatively correlated with tensile strength，strongly correlated with adhesive strength，weakly correlated with extensibility，stem ratio，stem binding force，and not correlated with other indicators；The rectangularity，slenderness，and contour temperature of tobacco had no correlation with branch adhesion，roundness had no correlation with stem ratio，while other mechanical indicators had certain impact on them. In summary，tobacco leaves from Chenzhou，Hunan had strong processing resistance and a relatively larger overall area.Tobacco leaves from Bijie，Guizhou had good uniformity in area and were mostly irregular in shape.Tobacco leaves from Sanming，Fujian had strong stem and branch binding force，but the overall area was relatively smaller.

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