Journal of Henan Agricultural Sciences

Research Progress on Preparation and Application of Straw Wood Vinegar

ZHAO Shuheng, LI Zhuojie, LIU Xiaofeng, HU Jianjun, DING Yanbo

2022, 51(1): 1-10. DOI: 10.15933/j.cnki.1004-3268.2022.01.001

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Wood vinegar，as a valuable acidic by‐product in the process of biomass pyrolysis for carbon production，is rich in various organic compounds such as acids，alcohols，phenols，aldehydes，esters，ethers and a small amount of metal elements，which has a high application value.In the raw material of wood vinegar，because of low lignin content and poor quality in straw，the development and application of straw wood vinegar are slow.However，the study of straw wood vinegar is of great significance to the utilization of agricultural waste and has a broad development prospect.In order to provide references for the further development and efficient utilization of straw wood vinegar，the crude preparation and refining technology of straw wood vinegar were introduced，the composition and physicochemical properties of straw wood vinegar were summarized，and the research and application progresses of straw wood vinegar in agriculture，medicine and health and other fields were reviewed.

Research Progress on the Effects of Biochar on the Growth of Medicinal Plants

ZHOU Ying, ZHOU Xinwei, ZHENG Ting, XIONG Pengfei, ZHANG Jiachun

2022, 51(1): 11-20. DOI: 10.15933/j.cnki.1004-3268.2022.01.002

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Based on the research on the application of biochar in the cultivation of medicinal plants at home and abroad，this paper summarizes the physical and chemical properties of 7 kinds of biochar used in the cultivation of medicinal plants，and systematically expounds the effects of biochar on the physical and chemical properties of soil，the form and content of nitrogen，phosphorus and potassium nutrients，the availability of heavy metals，enzyme activity and microbial diversity，as well as the growth and chemical composition content of medicinal plants.The mitigation effects of biochar on continuous cropping obstacles，fluoride toxicity and waterlogging are reviewed. The application prospect and future research direction of biochar in medicinal plant cultivation are put forward.

Preliminary Study on Mechanism of Auxin and Strigolactone Involved in Phosphate‐Modulated Tillering of Rice

ZHAO Chenyun, GUO Xiaoli, CHEN Yake, GONG Siyuan, LI Junzhou, ZHAO Quanzhi, SUN Huwei

2022, 51(1): 21-26. DOI: 10.15933/j.cnki.1004-3268.2022.01.003

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The changes of tiller number and tiller bud length of rice under low phosphate（LP，10 μmol/L）and normal phosphate（NP，300 μmol/L） conditions were studied，and the effect of application of exogenous auxin （naphthylacetic acid，NAA，10 nmol/L） and auxin polar transport inhibitor（N‐1‐naphthylphalamic acid，NPA，0.5 μmol/L） on tillering of wild type and mutant of D14（key strigolactone signal transduction gene）was explored，so as to reveal the mechanism of rice tillering in response to LP stress and the relationship between strigolactone and auxin in this process. The results showed that，compared with NP treatment，the tiller number and tiller bud length of LP treatment significantly reduced. The application of NAA inhibited tillering of rice under NP condition，and the application of NPA could not promote tillering of rice under LP condition. The mutant d14 was insensitive to phosphate，and had more tillers under LP and NP conditions，suggesting that D14 played an important role in phosphate‐modulated tillering of rice. The application of NAA significantly reduced tiller number of mutant d14 under LP and NP conditions. In conclusion，both auxin and strigolactone are involved in LP‐inhibited tillering of rice，but they may have a certain independence in this process.

Role of Laccase Gene OsLAC6 in Response to Arsenite Stress in Rice

XU Yiqun, GUO Yao, LIU Qingpo

2022, 51(1): 27-33. DOI: 10.15933/j.cnki.1004-3268.2022.01.004

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The expression pattern of laccase gene OsLAC6，as well as laccase activity，lignin content in the roots，phenotypes of arsenite tolerance，and the changes of contents of chlorophyll and H₂O₂ of wild type（WT）and transgenic rice lines overexpressing OsLAC6 gene under arsenite stress were examined，so as to uncover the role of OsLAC6 gene in response to arsenic stress in rice.The results showed that the OsLAC6 gene exhibited apparently temporal and spatial expression pattern，which was mainly expressed in the roots，stems，leaves and nodes of rice plants at tillering stage and roots，stems leaves of rice plants at mature stage.Overexpression of OsLAC6 gene extensively significantly increased the laccase activity，but decreased lignin content in roots and the arsenite tolerance of transgenic rice lines.Under arsenite treatment，transgenic rice lines overexpressing OsLAC6 gene exhibited significantly shorter relative root elongation，lower chlorophyll content in leaves，and higher H₂O₂ content compared with WT.Therefore，OsLAC6 negatively regulates the arsenite tolerance in rice.

Optimization of CRISPR/Cas9 Genome Editing Systems in Protoplasts of Setaria italica

LIU Guangyu, XU Xiaojing, XIA Keke, SUN Haixi, TAO Yueru, CUI Zhen, GU Ying

2022, 51(1): 34-42. DOI: 10.15933/j.cnki.1004-3268.2022.01.005

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To optimize and screen efficient genome editing systems in millet（Setaria italica），multiple CRISPR/Cas9（clustered regularly interspaced short palindromic repeats/CRISRP‐associated nuclease 9）genome editing systems were constructed with six kinds of gRNAs designed for phytoene desaturase（SiPDS） gene（gRNA1—gRNA2 for exon 1，gRNA3—gRNA6 for exon 12），and transformed into protoplasts of millet by polyethylene glycol‐mediated method. Then the mutation efficiency against SiPDS gene was rapidly detected by large‐scale parallel sequencing. The results showed that the protoplast transformation system established by the young stems of 7 day‐old millet etiolated seedlings had high transformation efficiency from 50. 44% to 57. 36%. Using this method，the gene editing systems with Cas9 gene driven by Super and Ubi promoters were transferred into millet protoplasts，the mutation efficiencies of SiPDS gene were 0. 5% and 5. 5% respectively. Thus，the Cas9 gene of gene editing systems with single gRNA，double gRNA or tRNA cassettes were driven by Ubi promoter，and mutation efficiency of SiPDS gene induced by these systems was compared. It was found that the mutation efficiencies of gene editing system with double gRNA cassettes such as Ubi‐dgRNAE1 and Ubi‐dgRNAE12 were 1. 66 and 1. 11 times higher than that with single gRNA system. The mutation efficiency induced by tRNA‐based system Ubi‐tRNA was 51. 24%，which was 5. 87 times higher than that with single gRNA system. Moreover，Ubi‐tRNA could simultaneously edit multiple sites of SiPDS gene，the multi‐site mutation frequency was 3. 23%. The cleavage activities in vitro of ribonucleoprotein（RNP）complexes which were mixture of gRNA3，gRNA4 or gRNA5 transcripts with Cas9 protein respectively were compared，it was found that RNP‐gRNA5 complex had the highest cleavage activity in vitro，the mutation efficiency of SiPDS gene was 2. 0%，and the dominant mutation type was deletion less than 3 bp.

Effects of Different Tillage Methods on Wheat Grain Quality under Fertilization and Non‐fertilization Conditions

NIE Shengwei, ZHANG Qiaoping, XU Jidong, ZHANG Yuting, ZHENG Nian, PAN Xiuyan

2022, 51(1): 43-49. DOI: 10.15933/j.cnki.1004-3268.2022.01.006

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The effects of vertical rotary tillage［FL，（30±5）cm］and traditional rotary tillage［XG，（12±5）cm］on wheat grain yield，quality and dough rheological properties were studied under fertilization（F）and non‐fertilization（F0）conditions in two consecutive wheat seasons（2017—2018 and 2018—2019），so as to provide reference for high‐quality and green wheat production. The results showed that the yield of FL treatment increased by 2.35%—34.13% compared with XG treatment，and the grain protein content and bulk density of FL treatment increased too，which were significantly increased by 17.51%and 0.87% under F0 condition in 2019 respectively. Compared with XG treatment，grain water content，wet gluten content，water absorption of flour，stability time of dough，energy，constant deformation tensileresistance，extensibility and maximum tensile resistance of FL treatment increased in 2018 and decreased
in 2019 under F0 condition；softening degree of dough decreased in 2018 but significantly increased in 2019；the flour extraction rate significantly decreased by 1.52%（in 2018） and 3.35%（in 2019）respectively；development time of dough shortened，and significantly shortened by 17.86% in 2018.Under F condition，the grain water content，flour extraction rate and stability time of dough of FL treatment increased；softening degree of dough decreased or kept equal，wet gluten content and extensibility decreased in 2018 and increased in 2019；water absorption of flour，development time of dough，energy，constant deformation tensile resistance，maximum tensile resistance decreased.In conclusion，vertical rotary tillage can obtain higher yield，increase grain protein content and bulk density，and improve or increase some rheological properties under non‐fertilization in the first season，but there is no obvious regularity under fertilization condition.

Analysis of Yield and Quality of Soybean Varieties in Regional Trial in Henan Province from 1985 to 2020

LI Jinhua, SHU Wentao, CHANG Shihao, YANG Qingchun, ZHANG Donghui, ZHANG Baoliang, GENG Zhen

2022, 51(1): 50-59. DOI: 10.15933/j.cnki.1004-3268.2022.01.007

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To study the trend of changes in the yield and quality of soybean varieties approved by Henan Province，we analyzed yield，protein and oil content in regional test of 126 soybean varieties approved by Henan Province from 1985 to 2020. The results showed that the highest yield of certified soybean varieties was 3 438.75 kg/ha，the lowest was 1 973.70 kg/ha，and the average yield was 2 789.40 kg/ha.The average yield of annual certified varieties in regional trials was 1 995.53—3 339.50 kg/ha，and the overall trend was increasing. The protein content of certified soybean varieties was 33.25%—50.18%，with an average of 42.89%，and the average protein content of annual certified varieties was 39.16%—47.29%，showing an overall trend of decreasing. The fat content of the certified soybean varieties was 17.10%—22.85%，with an average of 19.75%.The average fat content of annual certified varieties was17.80%—21.87%，showing an upward trend.The sum of protein and fat content was 50.35%—73.03%，and the sum of protein and fat content of annual certified varieties was 56.96%—69.16%，showing a slow downward trend. There were 25 high‐protein（≥45%）soybean varieties and 14 high‐oil（≥21.5%）soybean varieties among the 126 soybean varieties registered in Henan Province. Correlation analysis showed that soybean yield was extremely significantly negatively correlated with protein content，with a correlation coefficient of -0.310**，and was extremely significantly positively correlated with fat content，with a correlation coefficient of 0.357**. In the future，soybean breeding in Henan Province should pay attention to improvement of the quality based on increasing the yield of varieties to make the coordinated development of yield increase and high quality.

Effects of Straw Microorganisms Returning on Soil Nutrients，Rhizosphere Soil Bacterial Community Diversity and Yield of Winter Wheat

2022, 51(1): 60-70. DOI: 10.15933/j.cnki.1004-3268.2022.01.008

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To explore the effects of straw microorganisms entering soil on the rhizosphere microbial diversity and yield formation of the next crop，corn straw was used as the experimental material，under potted test conditions，with no straw returning to the field（SR0）as the control，two treatments of straw sterilization returning（SRD）and straw direct returning（SR）were designed，and the effects of different treatments of straw on soil nutrient content，rhizosphere bacterial community diversity and winter wheat growth were studied. The results showed that，straw returning promoted the contents of soil organic matter and alkaline nitrogen，with no significant differences between SRD and SR treatments. SR significantly increased the contents of soil available phosphorus and available potassium，but SRD decreased the contents of soil available phosphorus and available potassium.The plant height，tillering number，dry matter accumulation and yield components of wheat were all increased after straw returning. Among them，the plant height，tillering number，dry matter accumulation，effective panicles and yield were increased more in the SR treatment than in the SRD treatment.A total of 1 878 opentional taxonomic units of bacteria were detected in the samples，belonging to 23 phyla，81 classes，128 orders，223 families and 373 genera.The total OTUs，richness index and diversity index of wheat rhizosphere soil bacteria in the three treatments showed SR>SRD>SR0，with no significant differences among the three treatments.The analysis of the species community composition showed that Proteobacteria，Acidobacteria，Actinobacteria，Bacteroidetes，Gemmatimonadetes and Chloroflexi were the dominant phyla of rhizosphere soil bacteria，accounting for more than 90% of all bacterial communities. Under the class classification level，Alphaproteobacteria，Subgroup_6，Actinobacteria，Gammaproteobacteria and Sphingobacteria were the dominant classes of rhizosphere soil bacteria. The results of principal coordinate analysis showed that principal component 1（PC1）and principal component 2（PC2）explained 47.11% and 14.27%，and there were significant differences in the composition of rhizosphere bacterial communities among the three treatments. Redundancy analysis revealed that the first two ranking axes together explained 42.79% of the variation in the bacterial community. Soil organic matter content and alkaline nitrogen content were important factors affecting the bacterial community.In conclusion，straw microorganisms significantly promoted soil nutrient content，rhizosphere soil bacterial diversity and winter wheat yield under the condition of straw returning，so straw direct returning with microorganisms had a better effect.

Analysis of Histone H4 K16 Acetylation in Genome‐Wide Promoter Region in Saccharomyces cerevisiae under Acetic Acid Stress

MA Tian, WANG Yujing, CHENG Shuang, ZHANG Xiaohua, LIU Xiangyong

2022, 51(1): 71-78. DOI: 10.15933/j.cnki.1004-3268.2022.01.009

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For the exploration of new epigenetic regulation mechanism of yeast tolerance to acetic acid，changes of the histone H4 K16 acetylation modification in genome‐wide promoters in Saccharomyces cerevisiae under acetic acid stress were analyzed using the chromatin immunoprecipitation‐chip technique（ChIP‐chip）. Results showed that the acetylation levels of histone H4 K16 of 73 yeast genes in the promoter region changed in contrast to those in the control group after acetic acid（60 mmol/L for 30 min）stress treatment. A total of 19 genes with increased histone H4 K16 acetylation levels were mainly distributed on chromosomes 2，4，5，7，13，15 and 16，and 54 genes with decreased histone H4 K16 acetylation levels were mainly distributed on chromosomes 1，2，4，5，6，7，8，10，11，12，13，14，15 and 16.Gene function analysis showed that the genes were mainly concentrated in base‐excision repair，cytoskeleton organization，cell cycle process，and protein‐containing complex assembly，classified by biological process；The genes were mainly concentrated in vacuolar proton‐transporting V‐type ATPase，DNA polymerase complex，proton‐transporting two‐sector ATPase complex/proton‐transporting domain，mitotic spindle，and nuclear chromosome，classified by cellular component；The genes were mainly concentrated in the single‐stranded DNA 3′—5′ exodeoxyribonuclease activity，proteasome binding，and ATPase activity/coupled to transmembrane movement of ions，classified by molecular function.Three genes（ATP12，VPS55，and DLT1） with histone H4 K16 acetylation alterated in their gene promoter regions were randomly selected for chromatin immunoprecipitation‐quantitative real‐time PCR detection，and the results were consistent with the ChIP‐chip analysis，verifying the accuracy of ChIP‐chip data.Overall，these results indicate that the acetylation of histone H4 K16 in the gene promoter region is closely related to acetic acid tolerance in Saccharomyces cerevisiae.

Isolation and Compost Application of High‐Activity Cellulose‐degrading Strains from Distiller’s Grains

WU Yaoling, XI Xiaoli, ZENG Xianglian, LÜ Xibin, WANG Heyu, WANG Li

2022, 51(1): 79-88. DOI: 10.15933/j.cnki.1004-3268.2022.01.010

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In order to efficiently degrade cellulose in distiller’s grains，promote the rapid composting of distiller’s grains，and improve the quality of lees organic fertilizer，cellulose‐congo red medium and Hutcherson filter paper strip liquid medium were used to isolate high‐activity cellulose‐degrading strains from distiller’s grains. The results showed that three high‐activity cellulose‐degrading strains（MM2，MM6 and MX8）were obtained from distiller’s grains，which were identified as Paecilomyces varioti，Aspergillus fumigatus and Bacillus licheniformis，respectively.The results of straw‐degrading test showed that there was a natural synergy between any two of the three strains in cellulose degradation.Three kinds of strains were inoculated respectively on the bran to propagate，and then mixed to prepare a complex microbial community（CMC）according to the ratio of 1∶1∶1，and 0.6%，0.8% and 1.0% of CMC were added to the distiller’s grains to verify the compost function of CMC. The results showed that the CMC could quickly colonize and grow in the high‐acid environment of distiller’s grains，and had strong adaptability to the acid environment. The addition of CMC during the composting process of distiller’s

Variation of Microbial Flora in Vermiculite Matrix after Synergistic Control of Potato Common Scab Disease

WU Xiaoying, YANG Fangyan, LI Shouru, JIA Jingli, YU Xiumei, LIU Daqun, ZHAO Weiquan

2022, 51(1): 89-96. DOI: 10.15933/j.cnki.1004-3268.2022.01.011

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In this study，four treatments with different control effects［bacterial agent ZWQ‐1（abbreviated as LNBX1，with control effect of 33. 26%），fludioxonil+ZWQ‐1（LNBX3，72. 60%），thiram+ZWQ‐1（LNBX5，57. 92%）and tetramycin（LNBX8，24. 75%）］and one water treatment（control）were set up，and their vermiculite matrix were quantitatively sampled and analyzed by Illumina MiSeq high‐throughput sequencing to explore the correlation between the change of microbial community structure and the occurrence and development of potato scab. The results showed that the bacterial abundance and diversity of the control treatments in vermiculite substrates were lower than the water control. The LNBX3 and LNBX5 treatments with better control effects had higher fungal diversity than the control，while the LNBX1 and LNBX8 treatments with poorer control effects were lower than the control. After comparing the number of bacterial groups at different classification levels，it was found that bacteria of LNBX3 and LNBX5 treatments increased slightly at the species level compared with the control，while the number of groups at phylum，class，order，family and genus levels showed a decreasing trend；LNBX1 and LNBX8 treatments showed a decreasing trend in the number of bacteria at all classification levels. For the fungal groups number in vermiculite substrates，the analysis result showed that all of the four contol treatments increased at the genus and species levels compared with the control. The best control treatment LNBX3 was selected to analyze the composition and abundance of the dominant flora，and it was found that the abundance of Proteobacteria，Basidiomycota，Glomeromycota，Chytridiomycota and Rozellomycota increased，among which the abundance of Proteobacteria increased by 31.8%，whereas the relative abundance of Actinobacteria and Ascomycota decreased by 18.3% and 48.1%，respectively.The relative abundance of Pseudomonas，Bradyrhizobium，Burkholderia，Rhodanobacter，Archaeorhizomycetales and Chaetomium increased，while the relative abundance of Pedobacter，Mycoplana，Streptomyces，Monographella，Acremonium and Aspergillus decreased in varying degrees. The results indicate that the synergistic control with biocontrol bacteria and pesticide can significantly affect the composition of microbial flora in minituber vermiculite matrix，and the abundance of some beneficial microorganisms，including Pseudomonas，Brachyrhizobium，Burkholderia，Chaetomium，is increased.

Biogeographical Regionalization of Phycitinae in China Based on Multivariate Similarity Clustering Analysis

YANG Linlin, LIU Hongxia, REN Yingdang

2022, 51(1): 97-106. DOI: 10.15933/j.cnki.1004-3268.2022.01.012

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The purpose of this study was to propose the distribution pattern and summarize distribution characteristics of the subfamily Phycitinae in China. The distribution pattern of Phycitinae in China was analyzed based on bibliographical materials of 107 genera and 480 species by using multivariate similarity clustering analysis（MSCA）.Fauna components and distribution pattern of the subfamily were also discussed. The result showed that the total similarity coefficient of 64 basic geographical units in China was 0.100，and at the similarity level of 0.200，eight unit groups were clustered to represent eight geographical regions in China：Northwestern region，Northeastern region，Northern region，Changjiang‐Huaihe region，Central region，Southwestern region，Eastern region and Southern region；the Chinese Phycitinae fauna was comprised of 20.83% East Palaearctic component，4.79% Northwest Palaearctic component，26.88% Indo‐Pacific component，and 47.50% Eurytopic component，among which the transitional component consisting of East Palaearctic+Indo‐Pacific（23.54%）was the main component.In the world entomogeographical divisions，Chinese Phycitinae was mainly distributed in the Indo‐Pacific kingdom，followed by the East Palaearctic kingdom at genera level，and at species level，they mainly distributed in the East Palaearctic kingdom，followed by the Indo‐Pacific kingdom.In the Chinese geographical divisions of Phycitinae，the Northwestern China region showed lower genera/species richness but higher uniqueness，Northeastern China and Northern China regions showed similar genera/species richness and higher commonness，Changjiang‐Huaihe region showed the lowest uniqueness，which indicated a transitional region，Southern region showed both the highest genera/species richness and uniqueness，Eastern China region showed higher genera/species richness but lower uniqueness，Central China region showed lower uniqueness，which had higher commonness with Eastern China region，and Southwestern China region showed higher commonness with Southern China region. It is concluded that the distribution pattern of Phycitinae is similar to that of Insecta in China，but with some differences，which are caused by insufficient investigation and the much smaller sample of Phycitinae than that of Insecta；the main fauna components of Phycitinae in China are Palaearctic，Indo‐Pacific and their transitional groups；the distribution of Phycitinae in China is not uniform，faunas with high similarity in ecological environment have high similarity in genera and species distribution，and vice versa；the changes of the faunal components among regions are continuous，the farther the geographical distance，the greater the difference.

Photosynthetic Characteristics and Yield Components of Different Tree Crowns of Armeniaca sibirica

ZHANG Jian, CHEN Jianhua, SUN Yongqiang, DING Shifu, ZHENG Yongtao, DONG Shengjun

2022, 51(1): 107-115. DOI: 10.15933/j.cnki.1004-3268.2022.01.013

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In order to reveal the spatial variation of photosynthetic characteristics and yield of different crown of Armeniaca sibirica，and to filter the types of high yield crown of A.sibirica，this paper used A.sibirica as experimental materials based on the probability classification of branch angle and dividing tree shapes，measured the photosynthetically active radiation（PAR），net photosynthetic rate（Pn），transpiration rate（Tr），stomatal conductance（Gs）and yield of different tree shapes，canopies of A. sibirica and analyzed the diurnal changes of photosynthesis and yield differences. The results showed that A.sibirica was divided into compact shape（α<30°），uniform shape（30°≤α≤39°），and open shape（α>39°）based on the branch angle.The Pn，Tr，and Gs of different tree shapes，canopies and growth periods mainly displayed June>July，September，outer>middle>inner，open shape>uniform shape>compact shape.The diurnal variation of Pn of open shape showed unimodal curve in each month. The Pn of each canopy of uniform shape showed unimodal curve in June and September and bimodal curve in July.The Pn of each canopy of compact shape showed unimodal curve in June and bimodal curve in July and September.Among different canopies，the max of Pn of the outer canopy of different tree shapes was extremely significantly higher than the inner canopy，except that the max of Pn of compact shape outer was not significantly different with its inner in July，and the max of Pn of uniform shape outer was significantly higher than that in the inner in September. Among different tree shapes，the max of Pn of open shape was extremely significantly or significantly higher than other tree shapes in June.In July，the max of Pn of open shape was higher than other tree shapes，and there were significant differences in the middle and outer.However，in September，there was no significant difference in max of Pn between the different tree shapes. For Tr，except that different canopies of open shape in June and different canopies of compact shape in September were bimodal curve，different canopies of other tree shapes showed unimodal curve.For Gs，the canopies with different tree shapes showed unimodal curve in June and bimodal curve in July and September.The highest yield of 1 006.88 g/plant was found in the outer canopy，accounting for 53.61%，which was extremely significantly higher than in the middle canopy or inner canopy. The yield of open shape（1 303.00 g/plant）was extremely significantly higher than the uniform shape and compact shape.

Effect of Low Temperature Treatment on Nuclear Morphology and Physiological Indexes of Peony Petals

CHEN Yue, LIU Huichao, JIA Wenqing, HE Songlin, GUO Yingzi, ZHU Xiaopei, WANG Yanli, DU Xiaohua, WANG Zheng

2022, 51(1): 116-124. DOI: 10.15933/j.cnki.1004-3268.2022.01.014

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To analyze the cytological mechanism and physiological reasons of peony cut flower senescence at different temperatures，the effect of（25±1）℃（room temperature，contrast），2℃，4℃ and 8℃ on the vase life of peony was studied.The changes of nucleus morphology were observed and analyzed by DAPI，and the variations of protective enzyme（SOD，POD，CAT）activities，malondialdehyde（MDA），soluble protein content and superoxide anion（O-·2）generation rate during the process of flower arrangement in vase were analyzed.The results showed that the longest cut flower life was 17 d at 4，2℃.The order of the effect of different temperature treatments on the vase life of cut flowers was 4℃ =2℃ >8℃ .Low temperature treatment affected the normal rate of nucleus of cut flowers to different degrees. The normal rate of nucleus was basically 4℃>2℃>8℃，and the normal rate of nuclear could be used as an index to measure the health of cut flowers.The activities of SOD，CAT and POD of petals maintained a relatively high level，the petals could effectively remove excess reactive oxygen species and free radicals，and the metabolism was basically balanced，which was the physiological reason for the longer lifespan of cut flowers at 4℃.

Effect of Herbal Feed Additive as Dietary Supplement on the Quality and Nutritional Value of Gushi Chicken in Late Laying Period

ZHANG Yihui, ZHAO Xinlong, FU Yawei, KANG Xiangtao, LI Xiaofa, LI Donghua, JIANG Ruirui

2022, 51(1): 125-133. DOI: 10.15933/j.cnki.1004-3268.2022.01.015

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The purpose of this study was to explore the effects of Mengsheng herbal feed additive（Mengsheng grass for short）as a dietary supplement on carcass quality，meat quality and serum index of Gushi spent laying hens，so as to improve the economic value of spent laying hens. Six hundred 42‐week‐old Gushi chickens with similar body weight were randomly divided into two groups with 6 replicates in each group and 50 chickens in each replicate. Chickens in the control group were fed a basal diet，those in the experimental group were fed a basal diet supplemented with 1% Mengsheng grass，and the feeding period lasted for 45 days.The results showed that compared with the control group，the dressed percentage，percentage of half‐eviscerated yield and percentage of eviscerated yield of the experimental group increased by 0.58，1.99 and 0.93 percentage points respectively（P>0.05），the pH value of thigh muscle increased significantly at 45 min，and the drip loss of pectoral muscle and thighmuscle decreased by 1.18 and 1.55 percentage points respectively（P<0.05）.There was no significant difference in amino acid composition between the experimental group and the control group（P>0.05）.Compared with the control group，lignoceric acid（C24∶0）and docosahexaenoic acid（C22∶6n3）of the experimental group decreased by 20.75% and 17.21% respectively（P<0.05），the proportion of unsaturated fatty acids in total fatty acids increased by 3.59 percentage points，and the proportion of saturated fatty acids in total fatty acids decreased by 3.59 percentage points（P<0.05）. The Low density lipoprotein（LDL）of the experimental group decreased by 18.28% compared with the control group（P<0.05）. In conclusion，the diet supplemented with Mengsheng grass could improve the slaughter performance of Gushi chickens in the late laying stage to a certain extent，effectively improve the lipid metabolism，increase the fat content and tenderness of thigh muscle，reduce drip loss，optimize fatty acid composition，and then improve meat quality.

Effects of Different Additives on Silage Quality of Stem and Leaf of Tagetes erecta L

HOU Zhijiang, LIU Yanpei, CAI Ming, GAO Yuee, ZHANG Jicai, WANG Ling, LIU Jianyong, HUANG Bizhi

2022, 51(1): 134-140. DOI: 10.15933/j.cnki.1004-3268.2022.01.016

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The effects of lactic acid bacteria，glucose and formic acid additives on fermentation quality and nutritional components of marigold stem and leaf silage were studied in this experiment to comprehensively evaluate their regulation effect.Four treatment groups were set up with control group without any additives，lactic acid bacteria group was added at 5.0×106 cfu/g，glucose group was added at 2 g/kg，and formic acid group was added at 6 mL/kg，respectively. Samples were taken at 7 time points on the 3rd，6th，9th，12th，1th，30th and 45th days of silage to analyze the changes of dry matter and water soluble carbohydrate，the pH，contents of crude protein，acid washing fiber，organic acid，ammonia nitrogen and other indexes were analyzed after 45 days of ensilage. The results showed that with the extension of ensilage time，the dry matter and water soluble carbohydrate content of the lactic acid bacteria group and the glucose group decreased significantly（P<0.05），while dry matter and soluble carbohydrate content were lost less in the formic acid group and the control group due to slow fermentation（P>0.05）. After ensilaged for 45 days，the crude protein content of the lactic acid bacteria group and the glucose group was significantly higher than that of the control group and the formic acid group（P<0.05），while the ratio of neutral detergent fiber，acid detergent fiber and ammonia nitrogen to total nitrogen were significantly lower than that of control group and formic acid group（P<0.05）.The content of lactic acid in lactic acid bacteria group was significantly higher than that in other treatments（P<0.05），the content of acetic acid in formic acid group was significantly lower than that in other treatments（P<0.05），and no propionic acid and butyric acid were produced in all experimental groups. In conclusion，the three additives can improve the silage quality of marigold stems and leaves，among which lactic acid bacteria and glucose have the best effect，however，all the sensory quality cannot reach the first grade due to the biological characteristics of marigold.

Research on Linezolid Resistance and Mechanism in Clinical Isolates of Streptococcus suis

YANG Yingying, YI Kaifang, ZHANG Junkai, LUO Xingwei, LIU Peiyi, HAN Rongjia, HU Gongzheng

2022, 51(1): 141-145. DOI: 10.15933/j.cnki.1004-3268.2022.01.017

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AThe aim was to determine the resistance of Streptococcus suis clinical isolates to linezolid in seven provinces including Henan，Jiangxi，Hunan，Hubei，Anhui，Shanxi and Shaanxi，and analyze the mechanism of linezolid resistance. S. suis isolates collected from seven provinces in China between 2016 and 2019 were used as research subjects. BHI plates containing linezolid were prepared to screen linezolid‐resistant isolates. PCR method was used to detect linezolid resistance genes［optrA，cfr，cfr（B），cfr（C）and poxtA］.Antimicrobial susceptibility test was conducted by broth microdilution for the multidrug resistance of strains containing cfr or optrA genes. Results showed that 13 of 178 isolates were able to grow on the selective medium containing linezolid（4 mg/L）.The PCR results showed that all 13 linezolid‐resistant strains carried optrA gene，among which two strains carried cfr gene.The antimicrobial susceptibility test results showed that all cfr or optrA‐positive S. suis isolates were severely resistant to amikacin，gentamicin，erythromycin，azithromycin，bacitracin，doxycycline，clindamycin，florfenicol and chloramphenicol. In conclusion，all linezolid resistant strains carried optrA gene，which was dominant in mediating linezolid resistance，and all cfr or optrA‐positive S.suis stains were multiple resistant strains.Therefore，the routine surveillance of optrA gene of S. suis should be performed and the standard use of
antibiotics should be strengthened in pig industry.

Protective Effect and Mechanism of Proanthocyanidins B2 on Oxidative Damage in Porcine Granulosa Cells

XING Baosong, ZHANG Jiaqing, REN Qiaoling, LÜ Lingyan, WANG Xianwei, CHEN Junfeng, GAO Binwen, MA Qiang

2022, 51(1): 146-153. DOI: 10.15933/j.cnki.1004-3268.2022.01.018

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In order to study the protective effect of proanthocyanidins B2（GSPB2）on oxidative damage of porcine granulosa cells（PGCs），the model of the damage on PGCs induced by H2O2 was made by primary culture.At the same time，different concentrations of GSPB2 were used to intervene.The cellular apoptosis was detected by TUNEL method. The changes of intracellular ROS were observed by fluorescence staining. The activities of superoxide dismutase（SOD），glutathione peroxidase（GSH‐Px），catalase（CAT）and the content of malondialdehyde（MDA）were measured by chemochromatometry methods. The apoptosis related genes，such as Bcl-2，Bax，Fas，Puma and Sirt1 were detected by quantitative PCR.The senescence related proteins，such as Sirt1，p16 and p21 were detected by Western blot.The results showed that the cell apoptosis and ROS level increased significantly，SOD，CAT and GSH‐Px activities decreased，and MDA content increased（P<0. 05）after treatment with 200 μmol/L H₂O₂ for 12 h. Furthermore，the 10 μmol/L GSPB2 significantly alleviated the above conditions induced by H₂O₂.Further research found that the expression ratio of Bcl-2/Bax was up‐regulated，Puma and Fas expression were down‐regulated，Sirt1 mRNA and protein expression was up‐regulated，and the protein expression of p16 and p21 was down‐regulated.The above results indicate that GSPB2 has a protective effect on oxidative damage in PGCs，which could be related with the up‐regulation of Sirt1 protein，down‐regulation of p16 and p21 protein expression，increase in Bcl-2/Bax ratio，and the inhibition of Fas and Puma expression.

Estimation of Heading Time for Rice of Different Cultivars Based on UAV Remote Sensing Images

MO Jiacai, PENG Yi, LIU Xiaojuan, WANG Jing

2022, 51(1): 154-161. DOI: 10.15933/j.cnki.1004-3268.2022.01.019

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For the purpose of remotely monitoring and estimating heading time for rice of different cultivars，based on the multi‐temporal and multi‐spectral UAV remote sensing images of 1 102 rice cultivars under different climatic regions，we analyzed rice spectrum and vegetation index，fit the original multi‐temporal normalized difference red edge index（NDRE）data using single Gaussian function，and extracted the characteristic peak value of fitting curve. The results showed that the Pearson correlation coefficient between peak time and heading time reached 0. 906，based on which we established the remote sensing estimation model of multi‐cultivar rice heading time IHD_DAS=1.3×PEAK_DAS-18（DAS means the days after sowing，IHD DAS means DAS of initial heading date，PEAK_DAS means DAS of curve peak）.The model worked well for estimating the measured heading time.The root mean square error of 1 014 varieties in the modeling experiment was 3.15 d，and 5.02 d and 2. 99 d in the two verification experiments with 48 and 40 varieties，respectively. The model has good applicability to rice under different regions，climate and cultivars.

Research on Depth Model Compression Method Based on Joint Pruning for Seed Sorting

DONG Yan, LI Huanyu, LI Weijie, LI Chunlei, LIU Zhoufeng

2022, 51(1): 162-170. DOI: 10.15933/j.cnki.1004-3268.2022.01.020

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The crop seed sorting methods based on deep learning have achieved ideal recognition accuracy. However，their performance depends on the width and depth of the CNN（Convolutional neural network）model，which leads to a significant improvement in the number of model parameters，so it is difficult to deploy the larger‐scale model on the resource‐constrained edge devices. To address this issue，this paper proposes a depth model compression method based on joint pruning of channels and convolution layers.In the stage of channel pruning，the parameters of the BN layer trained by sparse regularization were used as the index to measure the importance of channels，to achieve the maximum channel pruning compression on the premise of keeping the accuracy of the model. Then，a layer pruning method based on a linear probe was proposed to compress the model and reduce memory access，to improve the reasoning speed of the model. Finally，the knowledge distillation technology was used in this paper to transfer the knowledge of pruning networks，thereby further compensating for the precision loss caused by pruning.The experimental results showed that，as far as the datasets of red kidney beans and maize seeds were concerned，the proposed method could improve the actual reasoning speed by 2.1 and2.8 times，while reducing the model calculation by 86. 55% and 91.55%. In the meantime，it still maintained good recognition accuracy（97.38% and 96.56%）.All these provide technical support for the deployment of the model in the actual seed sorting system.

Effects of Different Yellowing Treatments on the Quality of Flue‐cured Tobacco by Using Partial Least Squares‐Discrimination#br# Analysis

CHEN Feicheng, YANG Yide, DUAN Yufeng, LI Shengdong, YANG Yang, YAN Min, DUAN Weidong

2022, 51(1): 171-179. DOI: 10.15933/j.cnki.1004-3268.2022.01.021

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This research aims to study the effects of different yellowing methods on the quality of flue‐cured tobacco，and to clarify the appropriate yellowing method for flue‐cured tobacco.The effects of four types of yellowing methods，stage reduction of wet bulb，stress dehydration yellowing，medium temperature and medium humidity and high temperature moisturizing on the quality of flue‐cured tobacco leaves were determined，including the appearance and color，sensory quality，phenolic content，conventional chemical composition，free amino acid content and organic acid content. The partial least squares‐discrimination analysis（PLS‐DA）was used to comprehensively analyze the experiment results.Research indicated that，compared with other treatments，the color of flue‐cured tobacco treated with stress dehydration yellowing was the closest to the international orange color standard with better aroma characteristics，smoke characteristics，taste characteristics，and had the highest comprehensive sensory quality score（68.18）.In addition，the content of chlorogenic acid，rutin，nicotine，total nitrogen，potassium and proline of flue‐cured tobacco after the treatment of stress dehydration yellowing was the highest compared with other treatments. The PLS‐DA results showed that there were significant differences in the quality of tobacco leaves under the four yellowing methods，and nicotine，proline，and chlorogenic acid had higher variable importance in the projection（VIP），which could be used to improve tobacco quality as the key indicators of curing.

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