Journal of Henan Agricultural Sciences

Research Progress on Fine Mapping of QTLs for Yield Components of Rice

BAI Tianliang, LI Jie, RAN Jie, YANG Hui, QIAO Chengbin, LI Peifu, TIAN Lei

2022, 51(8): 1-8. DOI: 10.15933/j.cnki.1004-3268.2022.08.001

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Rice is one of major food crops，and its yield is very important for world food security.The yield components are key regulating factors for improvement of rice yield，which are often manifested as complex quantitative traits controlled by multiple genes. It is of great significance to explore favorable yield genes for high yield of rice. QTL mapping is an important method for identifying genes associated with excellent agronomic traits in crops.This paper elaborated the strategies of QTL fine mapping and population selection，reviewed the research progress on QTL fine mapping of three key yield components of rice，namely panicle number，grain number per panicle and grain weight，map⁃based cloning and functional analysis，and put forward the breeding strategy of rational utilization of rice yield component genes，so as to provide reference and theoretical basis for cloning excellent yield traits genes and unraveling the genetic mechanisms of yield traits in rice.

Identification and Analysis of WRKY Transcription Factors Differentially Expressed in Maize under Drought⁃Rewatering Treatment

FU Jiaxu, YAN Yali, XIE Xiaowen, ZHANG Xinyue, WEN Pengfei, GUAN Xiaokang, WANG Tongchao, WEI Li

2022, 51(8): 9-19. DOI: 10.15933/j.cnki.1004-3268.2022.08.002

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In order to explore WRKY transcription factors related to drought resistance in maize（Zea mays），differentially expressed WRKY genes were identified under drought⁃rewatering treatment，and their physicochemical characteristics，phylogenetic evolution，chromosome distribution，gene duplication，gene structure，conserved motif，cis⁃elements in promoter region and gene expression level were analyzed.The results showed that a total of 51 differentially expressed ZmWRKY genes were identified，the amino acid number，molecular weight and isoelectric point of 51 ZmWRKY proteins ranged from 99 to 729 aa，11.22 to 78.73 ku，and 4.58 to 12.26，respectively.ZmWRKY genes were divided into three groups Ⅰ，Ⅱ and Ⅲ，ZmWRKY genes of group Ⅱ were divided into Ⅱa，Ⅱb，Ⅱc，Ⅱd and Ⅱe.ZmWRKY genes were unequally distributed on 10 chromosomes，including 2 pairs of tandem duplication and 16 pairs of segmental duplication.The ZmWRKY genes contained 1—12 exons，most of ZmWRKY proteins（41）contained 2—4 conserved motifs，and WRKY members in the same group had similar motif compositions.Many cis⁃elements related to plant hormone and abiotic stress，such as ABRE，AuxRR⁃core，TCA⁃element，TC⁃rich repeats，TGACG⁃motif，LTR，MBS，TATC⁃box，P⁃box，CGTCA⁃motif，GC⁃motif，TGA⁃element and GARE⁃motif were identified in ZmWRKY gene promotor region.Under drought⁃rewatering treatment，51 ZmWRKY genes exhibited different expression patterns，among them，fifteen genes positively responded to drought stress，including ZmWRKY1，ZmWRKY10，ZmWRKY16，ZmWRKY28， ZmWRKY30， ZmWRKY33， ZmWRKY42， ZmWRKY65， ZmWRKY68， ZmWRKY78，ZmWRKY96，ZmWRKY99，ZmWRKY100，ZmWRKY102 and ZmWRKY111，and one gene negatively responded to drought stress，which were candidate genes for further study of drought response in
ZmWRKY family.

Effects of Arbuscular Mycorrhizal Fungi on Wheat Physiological Characteristics and Rhizosphere Soil Enzyme Activities under Cadmium Stress

GUO Hui, ZHOU Hui, ZHUANG Jingjing, WANG Xiaobing

2022, 51(8): 20-27. DOI: 10.15933/j.cnki.1004-3268.2022.08.003

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In order to explore the response of arbuscular mycorrhiza fungi（AMF）⁃wheat symbiote to Cd stress and provide theoretical basis for immobilization remediation in Cd polluted farmland，the photosynthetic and physiological characteristics，antioxidant enzyme activities and rhizosphere soil enzyme activities of wheat without AMF（CK），inoculated with Rhizophagies intraradices（Ri），inoculated with Funneliformis mosseae（Fm）and inoculated with Ri and Fm at 1∶1（RF）were studied under Cd stress［0 mg/kg（Cd0），5 mg/kg（Cd5），10 mg/kg（Cd10）and 20 mg/kg（Cd20）Cd］.The results showed that no matter whether AMF was inoculated，specific leaf weight，chlorophyll content，net photosynthetic rate（Pn）of wheat and rhizosphere soil enzyme activities decreased with the increase of Cd content，while malondialdehyde（MDA） content，catalase（CAT），superoxide dismutase（SOD） and peroxidase（POD） activities of wheat leaves increased. AMF inoculation could effectively improve photosynthetic and physiological characteristics，antioxidant enzyme activities and rhizosphere soil enzyme activities of wheat under Cd stress，and the effect was more significant under higher Cd content.RF treatment had the best improving effect on photosynthetic characteristics of wheat under Cd stress，but there was not significant difference in improving antioxidant enzyme and rhizosphere soil enzyme activities among different AMF treatments. Under Cd20，specific leaf weight，chlorophyll a content，chlorophyll b content and Pn of AFM treatment increased most by 14.56%，13.79%，13.68%，16.92% compared with CK respectively；leaf CAT，SOD，POD activities increased most by 33.69%，24.54%，14.99% respectively；soil sucrase（SC），CAT，urease（UE），dehydrogenase（DHA）activities increased most by 190.56%，185.66%，19344%，190.00% respectively；while MDA content decreased by 14.22%—17.30%.In conclusion，under Cd stress，AMF inoculation could effectively improve the photosynthetic and physiological characteristics，antioxidant enzyme activities and rhizosphere soil enzyme activities of wheat，and RF treatment was better in general.

Comparison and Comprehensive Evaluation of Quality of 20 Potato Varieties

ZHANG Fan, LI Yuan, CHEN Mengru, XING Yingying, WANG Xiukang

2022, 51(8): 28-36. DOI: 10.15933/j.cnki.1004-3268.2022.08.004

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The tuber yield per plant，starch content，reducing sugar content，soluble sugar content，vitamin C content and protein content of 20 potato varieties were analyzed，and the comprehensive evaluation of quality was done by principal component analysis，membership function method and cluster analysis，so as to screen out potato varieties with good quality and suitable for different processing purposes and cultivation in Yulin City. The results showed that there were great differences in quality indexes among different potato varieties.The variation coefficients of tuber yield per plant，starch content，reducing sugar content，soluble sugar content，vitamin C content and protein content were 46.76%，50.07%，50.66%，64.79%，33.72% and 68.10% respectively.The tuber yield per plant and starch content of Longshu 11 were the highest，the reducing sugar content of LZ112 was the highest，the soluble sugar content of Longshu 9 was the highest，the vitamin C content and protein content of Laohongpi were the highest.The results of principal component analysis showed that the quality indexes of 20 potato varieties could be expressed by four principal components（the cumulative variance contribution rate was 90.448%），and the representative indexes of each component were protein content，starch content，soluble sugar content and reducing sugar content respectively. Then，comprehensive evaluation was carried out by membership function method and cluster analysis.The results showed that seven varieties with better quality were screened out，accounted for 35.0% of the tested varieties，one variety（Dingshu 3）could be used for both vegetable and starch processing，one variety（Ganyinshu 1）could be used for whole flour processing and fried food processing，one variety（Longshu 11）could be used for starch processing after large⁃scale cultivation，two varieties（L08104⁃12 and Feurita）could be processed as whole flour，and two varieties（Jizhangshu 12 and Longshu 9）could be used for fried food processing.

Effect of Brassinolide on Root Architecture of Tobacco Seedlings and Regulation Mechanism

LIANG Dong, LIU Guangliang, SHI Yi, WANG Shusheng, WANG Chengdong, WANG Xiaolin

2022, 51(8): 37-44. DOI: 10.15933/j.cnki.1004-3268.2022.08.005

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In order to explore the regulation mechanism of brassinolide（BRs）on root architecture of tobacco seedlings，the differences of root architecture under different concentrations of exogenous EBR（2，4⁃epibrassinolide） and BRZ（brassinolide synthesis inhibitor） were investigated using the flue⁃cured tobacco Nongda 202.The changes of root tip cross⁃section of secondary lateral roots were observed by electron microscope，and the auxin content，GUS（β⁃glucosidase）staining and relative gene expression in lateral roots were determined to analyze the relationship between root architecture and auxin.The results showed that EBR could affected root length，root diameter and secondary lateral root density of tobacco seedlings.Lower concentration of EBR（0.1 nmol/L）increased the total root length by 29.0%，but higher EBR，such as 0.5，1，5，10 and 100 nmol/L declined the primary lateral root length by 4.6%，9.6%，18.5%，29.6% and 36.9% respectively.As the EBR concentration（0.1—10 nmol/L）increased，the density of secondary lateral roots increased，but the average root diameter gradually decreased.The average root diameter of tobacco increased significantly in BRZ treatment（100—500 nmol/L）.EBR affected the root diameter of tobacco seedlings mainly by regulating the number of tobacco lateral root cells and the diameter of column，the cross cutting diameters of secondary lateral root and column under 10 nmol/L EBR treatment were significantly reduced by 32.5% and 25.0% compared with CK（No EBR and BRZ）. The auxin content of lateral root decreased by 7.1% significantly under 10 nmol/L EBR treatment，but there was no change of lateral root auxin under 500 nmol/L BRZ treatment. EBR treatment increased the expression of PIN3aT，PIN3bS，PIN3bT and PIN11T in tobacco，which was considered to change the distribution of auxin in roots.Therefore，brassinolide can regulate the level of auxin in different parts of roots，and then affect the changes of tobacco root architecture.

Analysis of Flavor Characteristics and Microbial Community of Cigar Tobacco Leaves from Different Regions

2022, 51(8): 45-54. DOI: 10.15933/j.cnki.1004-3268.2022.08.006

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Twenty⁃four cigar tobacco leaves from three cigar tobacco producing areas，including Dominica，Indonesia and America，were used as experimental materials to investigate their flavor characteristics by sensory evaluation，and the microbial communities were determined by microbial high⁃throughput sequencing.By analyzing the differences in flavor and microbial community composition of cigar tobacco leaves from different regions，this study explored the relationship between microbial community and flavor characteristics of cigar tobacco leaves，and identified the key microorganisms in cigar tobacco leaves，to lay a foundation for the utilization of raw materials，product development and application innovation of domestic cigar industry. The results showed that Dominican cigar tobacco leaves had the flavors of nut，bean，wood，caramel，baked and hay；Indonesian cigar tobacco leaves had high honey and sour aromas，but also had more offensive odor；American cigar tobacco leaves had no prominent flavor and offensive odor.The dominant bacterial genera in cigar tobacco leaves from the three regions were Staphylococcus，Pseudomonas，and Sphingomonas，and the dominant fungal genera were Aspergillus，Sampaiozyma，and Alternaria.There were significant differences in microbial communities in tobacco leaves from different producing areas，and the difference in bacterial community in cigar leaves from different origins was significantly greater than that in fungal communities. Through the association network，it was found that bacteria Staphylococcus，Corynebacterium_1，Tetragenococcus and Atopostipes were closely related to the flavor of cigar tobacco leaves.The bacterial community had closer relationship with flavor characteristics than fungal community，and had an important contribution to the flavor formation of cigar tobacco leaves.

Comprehensive Evaluation of Wan Artemisia argyi Germplasm Based on Morphological traits and Quality traits

XU Lanjie, LIANGHuizhen, YUYongliang, YANG Qing, LI Lei, AN Sufang, YANG Hongqi, TANZhengwei, LIChunming, DONGWei, LU Dandan

2022, 51(8): 55-63. DOI: 10.15933/j.cnki.1004-3268.2022.08.007

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Scientific and reasonable methods were used to comprehensively evaluate and screen excellent germplasms of Wan A.argyi，so as to provide parental materials and technical support for the breeding of new varieties of Wan A.argyi.Thirty Wan A.argyi germplasms from Nanyang City were used as experimental materials to comprehensively evaluate their agronomic and quality traits by Shannon⁃weaver diversity index，membership function method，correlation analysis，principal component analysis and cluster analysis.The results showed that Wan A.argyi germplasms had high genetic diversity，the diversity index of different traits was 0.53—1.58，and the coefficient of variation was 16.95%—160.71%.Correlation analysis showed that most agronomic traits had significant or extremely significant correlations，and some quality traits had significant correlations，but the correlations between agronomic traits and quality traits were poor.Principal component analysis converted 15 traits into 5 comprehensive factors，with a cumulative contribution rate of 85.12%.The first，second and fourth principal components mainly reflected agronomic traits，the fifth principal component mainly reflected quality traits，and the third principal component mainly reflected agronomic traits and quality traits. Based on the square sum of deviation method，the 30 Wan A.argyi germplasms were clustered into three categories at the Euclidean distance of 14.0.The first category was high⁃quality type and included 8 germplasms；the second category was high⁃yield type，which contained 8 germplasms；the traits of germplasms of the third category were between the first and the second categories. Based on the membership function method，5 germplasms with excellent comprehensive performance in terms of yield and quality were screened out， which could provide parental material support for the breeding of new varieties of Wan A.argyi.

Genetic Diversity Analysis of Pseudostellaria heterophylla Based on ISSR and RAPD Molecular Markers

SHI Hui, CHEN Meixi, LI Pingping, YE Zuyun

2022, 51(8): 64-73. DOI: 10.15933/j.cnki.1004-3268.2022.08.008

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In order to explore the genetic diversity and genetic relationship of Pseudostellaria heterophylla germplasm resources，inter⁃simple sequence repeat（ISSR）and randomly amplified polymorphic DNA（RAPD） markers were used to analyze the genetic diversity of 10 Pseudostellaria heterophylla germplasms from different habitats. The results showed that 13 ISSR primers and 16 RAPD primers obtained 101 and 89 amplification sites in 10 germplasms of Pseudostellaria heterophylla.The number of polymorphism loci（NPL）were 58 and 63，and the percentage of polymorphism loci（PPL）were 57.37％and 70.32％ respectively.The average values of observed number of alleles（Na），effective number of alleles（Ne），Nei’s genetic diversity index（H） and Shannon’s information index（I） among Pseudostellaria heterophylla germplasms based on ISSR markers were 1.573 7，1.344 3，0.353 2 and 0.529 7，respectively.The average values of Na，Ne，H and I among Pseudostellaria heterophylla germplasms based on RAPD markers were 1.703 2，1.418 1，0.354 9 and 0.532 1，respectively.The GS between Pseudostellaria heterophylla germplasms ranged from 0.668 to 0.911 based on the two markers.The clustering results based on the synthesis of the two markers showed that 10 germplasms of Pseudostellaria heterophylla could be clustered into 4 groups when GS was 0.780，and the clustering results were significantly correlated with the geographical distribution of Pseudostellaria heterophylla germplasms.

Effects of Different Wheat Special Fertilizers on Yield，Economic Benefit and Nutrient Uptake of Wheat in Northern Henan

HU Feng, ZHANG Yunhong, MAO Jiawei, LIU Xiaoqi, QIAN Kai, ZHANG Jie

2022, 51(8): 74-83. DOI: 10.15933/j.cnki.1004-3268.2022.08.009

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Field experiments were conducted to study the effects of eight kinds of wheat special fertilizers （Woertian wheat special fertilizer，Xinlianxin wheat special fertilizer，Huatuo wheat special fertilizer，Duoweibao wheat special fertilizer，Jiashili wheat special fertilizer，Kebang wheat special fertilizer，common wheat special fertilizers and Yubei wheat special fertilizer）on the yield，economic benefits andnutrient uptake of wheat（Triticum aestivum L.）in Northern Henan，in order to provide a basis for the selection and rational application of wheat special fertilizers in Northern Henan.The results showed that，compared with the control without fertilization，the spike number and yield under the treatments of wheat special fertilizers were increased by 18.9%—41.8% and 17.6%—32.4% respectively，and those of Huatuo wheat special fertilizer and Duoweibao wheat special fertilizer treatments were relatively higher.Moreover，there were the highest output value，increased output value and net income in the treatment of Duoweibao wheat special fertilizer，which were 24 626.72，6 031.03 and 4 351.03 Yuan/ha respectively，and the output⁃input ratio of fertilization was also the highest，with value of 3.59.Among the eight kinds of wheat special fertilizers，total accumulation of nitrogen，phosphorus and potassium in wheat under the treatments of four kinds of wheat special fertilizers with high⁃nitrogen（N） and low⁃phosphorus（P）（Woertian wheat special fertilizer，Xinlianxin wheat special fertilizer，Huatuo wheat special fertilizer and Duoweibao wheat special fertilizer）（517.02—590.46 kg/ha）was higher than that of four kinds of wheat special fertilizer with low⁃N and high⁃P（481.76—502.77 kg/ha）（Jiashili wheat special fertilizer，Kebang wheat special fertilizer，common wheat special fertilizer and Yubei wheat special fertilizer），and the treatment of Duoweibao wheat special fertilizer had the highest fertilizer agronomy efficiency and partial productivity，with values of 8.66 kg/kg和35.38 kg/kg.The application of wheat special fertilizers increased the contents of alkali⁃hydrolyzed N，total and available P in soil after wheat harvest to different degrees，and those of common and Duoweibao wheat special fertilizer treatments were higher，increased by 150.0%，91.2%，71.6% and 114.3%，82.1%，54.9% compared with the control without fertilization.In general，the one⁃time application effects of high⁃N and low⁃P fertilizers were better than those of low⁃N and high⁃P fertilizers in maintaining high yield and efficient utilization of nutrients in this experiment，and Duoweibao wheat special fertilizer had the best application effects.

Isolation，Identification and Determination of Control Effect of Antagonistic Microorganisms against Sesame Fusarium Wilt

CHANG Shuxian, MA Qin, QU Wenwen, ZHANG Haiyang, MIAO Hongmei, DUAN Yinghui

2022, 51(8): 84-91. DOI: 10.15933/j.cnki.1004-3268.2022.08.010

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Sesame Fusarium wilt（SFW），caused by Fusarium oxysporum f.sp.sesami（FOS），is one of destructive fungal diseases in sesame. In order to screen antagonistic microorganisms for the biocontrol of SFW，the isolation，purification and antagonistic effect test of rhizospheric microorganisms were carried out by using root soil samples of sesame，hispid arthraxon and Chinese violet from different ecological regions in China. And the control effect of important antagonists on SFW at seedling stage was verified by pot experiment. Furthermore，the antagonistic strains were identified through morphological observation，physiological and biochemical characteristics assay and 16S rDNA sequence analysis.The results showed that a total of 1 077 bacteria，515 fungi，and 554 actinomycetes strains were isolated from 33 soil samples from six provinces in China，and 17 bacteria，10 fungi，and 51 actinomycetes were screened with antagonistic effect against F.oxysporum strains.Among them two bacterial isolates，SF3⁃33 and SF4⁃11 had the most significant antagonistic effect with maximum antagonistic distance being 1.0 cm，and their maximum control effects on SFW caused by the high pathogenic FOS strains reached 83.74% and 63.13% respectively.Further，SF3⁃33 was identified as Bacillus licheniformis，while SF4⁃11 was as Paenibacillus peoriae.In conclusion，a large number of rhizosphere microorganisms are screened，which can inhibit the mycelial growth of F.oxysporum.Among them，two bacterial strains SF3⁃33 and SF4⁃11 can significantly reduce the occurrence of SFW at seedling stage，which provides the important materials with application potential for further exploring the biological control technology of SFW.

Effect of Powdery Mildew Pathogens on Photosynthetic Characteristics and Chloroplast Ultrastructure of Pumpkin Leaves during Fruiting Stage

YE Jiajing, ZHAO Jinpeng, BIAN Shijie, GUO Weili, LI Xinzheng

2022, 51(8): 92-98. DOI: 10.15933/j.cnki.1004-3268.2022.08.011

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In order to explore the physiological effects of powdery mildew on pumpkin photosynthesis，the photosynthetic indexes，chlorophyll content，as well as cell necrosis and chloroplast ultrastructural changes were studied，with the high powdery mildew⁃resistant pumpkin inbred line 112⁃2 and the highly susceptible variety Jiujiang Jiaoding（JJJD） as test materials by spraying powdery mildew pathogens during the fruiting period.The results showed that compared with CK（without powdery mildew inoculation），the net photosynthetic rate（Pn），stomatal conductance（Gs），transpiration rate（Tr）and chlorophyll content of pumpkin leaves（112⁃2 and JJJD）inoculated with powdery mildew pathogens all decreased.The reduction in Pn of the resistant material was smaller than that of the susceptible material，which decreased by 45.4% and 63.8% at 2 days after inoculation respectively，and the total chlorophyll content decreased by 57.0% and 49.0% at 12 days after inoculation respectively，while intercellular CO2 concentration（Ci）increased.At 4 days after inoculation，necrotic spots near the infection point of both two pumpkin materials appeared，and chloroplasts were swollen and deformed，stromal lamella and basal lamella loosely arranged，and osmophilic bodies and starch granules increased. The area of necrotic spots and the degree of chloroplast deformation in JJJD were bigger than those of 112⁃2.It is concluded that the reduction in Pn of powdery mildew pathogens⁃infected pumpkin leaves at fruiting stage is mainly caused by cell necrosis occurrence，chlorophyll decrease and chloroplast structure destruction.The lesser decrease in Pn of the resistant material 112⁃2 is related to milder cell necrosis and chloroplast structure damage.

Analysis of Yield，Quality and Potassium Absorption and Utilization of Different Genotypes of Melon under Low Potassium Condition

KANG Liyun, LI Xiaohui, GAO Ningning, LIANG Shen, LI Hailun, WANG Huiying, CHANG Gaozheng, XU Xiaoli, ZHAO Weixing

2022, 51(8): 99-106. DOI: 10.15933/j.cnki.1004-3268.2022.08.012

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To explore the differences in yield，quality and potassium absorption of different genotypes of melon under low potassium condition，the field experiment was conducted with low potassium tolerant varieties（lines）Xuemi and Jiangjunyu and low potassium sensitive varieties（lines）T221 and T120 of melon as test materials. The differences of yield，quality，potassium accumulation and distribution and potassium utilization efficiency of melon under low potassium（0 kg/ha，LK）and appropriate potassium（300 kg/ha，CK）were studied. The results showed that，compared with CK，the yield，central and edge soluble solids，solid⁃to⁃acid ratio，soluble protein，and vitamin C content of T221 and T120 in low potassium treatment were significantly reduced（P<0.05） by 7.62%，15.85%，16.54%，26.24%，14.39%，23.51% and 10.22%，14.42%，17.82%，24.99%，18.97%，27.80%，respectively，and the titratable acid content had the opposite trend，with a significant increase of 12.86% and 14.00%（P<0. 05），while the differences of Xuemi and Jiangjunyu were not significant.Compared with CK，the potassium accumulation of stems，leaves，fruits and plants of T221 and T120 in low potassium treatment was significantly reduced（P<0.05）by 17.94%，20.32%，24.02%，21.49% and 20.58%，18.96%，23.26%，21.44%，respectively，while Xuemi and Jiangjunyu were not significantly different. Xuemi and Jiangjunyu significantly increased potassium utilization efficiency compared with T221 and T120（P<0.05），with an increase of 73.67%，while the potassium production efficiency and potassium fertilizer utilization rate were significantly reduced（P<0.05），which decreased by 21.76% and 55.37%，respectively，indicating that Xuemi and Jiangjunyu could absorb and utilize more potassium，obtain higher yield and quality，and transfer more absorbed potassium to the fruit under low potassium condition.Based on the above analysis，Xuemi and Jiangjunyu were not sensitive to potash fertilizer and were suitable for planting under low⁃potassium conditions.For low potassium tolerant varieties，potash input can be appropriately reduced for cost savings and pollution reduction，so as to achieve high⁃yield and high⁃efficiency cultivation of melon.

Effects of Spraying Ammonium Polyphosphate on Yield，Quality，Absorption and Transport of Cd in Pepper

CHAI Guanqun, LIU Guihua, LUO Muxinjian, WANG Li, JIANG Ya, FAN Chengwu

2022, 51(8): 107-116. DOI: 10.15933/j.cnki.1004-3268.2022.08.013

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In order to provide a scientific basis for the safe production of pepper in Cd⁃safely⁃utilized farmland and the development of foliar phosphate fertilizer，Cd⁃safely⁃utilized soil and the pod pepper variety‘Qing Hong Yuan Shuai’were selected as test materials，and a pot experiment was conducted to study the effects of foliar spraying of 0，0.1，0.2，0.5 g/L ammonium polyphosphate（expressed as CK，T1，T2，and T3，respectively）on the yield，quality，and Cd uptake and translocation of pepper.The results showed that with the increase of ammonium polyphosphate concentration on leaf surface，the yield of pepper fruit showed an increasing trend. Compared with CK，the yields of fresh fruit and dry fruit in T3 treatment increased by 11.11% and 11.53%，respectively.The contents of soluble sugar，soluble protein，amino acid，Vc and capsaicin in pepper fruits treated with T3 were significantly higher than those in CK.The Cd content（dry basis）in different parts of pepper was as follows：leaf on main stem>root>fruit>leaf on fork stem>fork stem>main stem.The bioaccumulation factor of pepper for soil Cd（BCF）reached 193.77%—351.82%，which decreased with the increase of ammonium polyphosphate concentration on leaf surface，and T3 treatment decreased by 158.05 percentage points compared with CK.The Cd content of pepper fruit decreased with the increase of ammonium polyphosphate concentration on leaf surface.The Cd content of pepper fruit in T3 treatment was significantly lower than that in CK，and the decrease rates of fresh fruit and dry fruit were 16.67% and 11.84%，respectively.Correlation analysis showed that‘mass dilution’and increasing Cd accumulation and retention ability of leaves on fork stem and main stem of pepper by foliar application of ammonium polyphosphate might be two main reasons for the decrease of Cd content in fruit. In conclusion，foliar spraying of 0.5 g/L ammonium polyphosphate has the best effect on yield and quality improvement and Cd reduction of pepper fruit.

Effects of Ultrasound on Seed Germination and Seedling Physiological Characteristics of Echinacea purpurea under Salt Stress

YIN Juan, ZHU Qingsong, ZHAO Jingjing, CAO Yongyi, MA Guihua

2022, 51(8): 117-127. DOI: 10.15933/j.cnki.1004-3268.2022.08.014

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E.purpurea seeds were taken as materials，seed germination and seedling physiological characteristics of E.purpurea under different salt stress（0，0.45%，0.90% NaCl）were studied by using ultrasonic pretreatment with different time（0，5，25，45 min），filter paper bed method and potted seedling growth experiments.The results showed that the germination of E.purpurea seeds was significantly inhibited under salt stress，and the higher the salt content，the stronger the stress inhibition.Seed germination rate，germination potential，germination index，vigor index，aboveground dry weight and root dry weight of seedlings，and the activities of superoxide dismutase（SOD），peroxidase（POD）and catalase（CAT）in seedling leaves were significantly improved after ultrasonic treatment.In addition，appropriate ultrasonic treatment under salt stress could alleviate the stress by adjusting the content of proline，soluble sugar and soluble protein in roots.Compared with 0 min（UaS1），under low salt（0.45% NaCl）stress，the content of proline and soluble sugar decreased by 27.23% and 12.23% respectively，and the content of soluble protein increased by 92.81%.Compared with 0 min（UaS2），under high salt（0.90% NaCl）stress，the content of proline and soluble sugar decreased by 11.43% and 12.75% respectively，and the content of soluble protein increased by 87.83%.In conclusion，proper amount of ultrasonic treatment can promote the seed germination and seedling growth of E.purpurea，and enhance the resistance of seedlings to salt stress.The optimum ultrasonic treatment time was 25 min.

Seed Germination Characteristics and Salt Resistance of Wild Leptodermis oblonga Bunge

GUO Li, SUO Sumin, YANG Hongli, XU Minghui, CHANG Jietian

2022, 51(8): 128-133. DOI: 10.15933/j.cnki.1004-3268.2022.08.015

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In order to develop local characteristic wild plants，seeds of Leptodermis oblonga Bunge were used as test materials，by setting different treatments，including culture temperature of 15，20，25，30，35 ℃ and variable temperature 15℃/25℃，buried depth of 0.5，1.0，1.5，2.0，2.5，3.0 cm，GA₃ mass concentration of 0.5，1.0，1.5，2.0，3.0 mg/L，NaCl solution of 50，100，150，200，250，300 mmol/L，seed germination and salt resistance during seed germination stage of Leptodermis oblonga Bunge were studied.The results showed that the best temperature for seed germination of Leptodermis oblonga Bunge was 25℃ ，and the variable temperature 15℃/25℃ was bad for seed germination. The best cover thickness of seeds was 1.0 cm，and no seedlings emerged under 2.5 cm soil. GA3 could promote seed germination，and 1.5 mg/L was the optimal concentration for germination of Leptodermis oblonga Bunge seeds.The seedlings of Leptodermis oblonga Bunge could normally grow under 50—100 mmol/L NaCl，while 150 mmol/L NaCl inhibited the growth of seedlings，indicating that the seeds had a certain ability to adapt to low⁃concentration NaCl stress，and high⁃concentration NaCl stress could cause plant death or germination failure.

Effects of Silage Mulberry Leaves on Production Performance and Meat Quality of Hu Sheep

MA Huihui, XU Bin, WEI Fengxian, LI Wenjia, SUN Quanyou, WANG Linyi, DENG Wen, FU Chen, LI Shaoyu

2022, 51(8): 134-142. DOI: 10.15933/j.cnki.1004-3268.2022.08.016

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This study was conducted to study the effects of silage mulberry leaves on growth performance，meat quality of Hu sheep. 160 healthy Hu sheep with average weight of（19.12±1.04）kg were randomly divided into four groups with five replicates in each group and eight sheep in each replicate.The control group（group Ⅰ）was fed with peanut seedlings，and the mulberry leaves and peanut seedlings were mixed at 15∶85（group Ⅱ），30∶70（group Ⅲ）and 45∶55（group Ⅳ）in treatment groups，respectively.The experimental period lasted for 60 days. The results showed that average daily feed intake，average daily gain，feed to gain ratio and slaughter performance indexes were not significantly different among the groups（P>0.05）.The muscle cooking loss and shear force of experimental groups Ⅱ，Ⅲ andⅣ were reduced by 17.14%，19.54%，16.57% and 7.92%，9.50%，6.75% compared with those in group Ⅰ，respectively. The protein content in the muscle of Hu sheep in groups Ⅲ and Ⅳ were increased by 9.31%and8.27% compared with that in group Ⅰ，respectively. The content of palmitic acid in the muscle of group Ⅲ was significantly lower than that of groups Ⅰ，Ⅱand Ⅳ.The contents of a⁃linolenic acid and arachidonic acid in muscle of groups Ⅲ，Ⅳ were significantly higher than those of group Ⅰ.The contents of saturated fatty acids in the muscles of groups Ⅱ，Ⅲ and Ⅳ were reduced by 6.00%，10.29% and 7.03% compared with that in group Ⅰ，respectively. The content of monounsaturated fatty acids in group Ⅲ was significantly higher than that in other groups，and the content of polyunsaturated fatty acids in group Ⅳ was significantly higher than that in other groups（P<0.05）.The contents of aspartic acid，glutamic acid，leucine，lysine，tryptophan，total amino acids，and flavor amino acids in the longissimus dorsi muscle of Hu sheep in groups Ⅲ and Ⅳ were significantly higher than those in groups Ⅰ and Ⅱ（P<0.05）.In conclusion，silage mulberry leaves have certain promoting effects on meat quality and contents of amino acid and unsaturated fatty acid in muscle of Hu sheep，and the supplementation of 30% mulberry leaves has the best effect.

Construction of Eukaryotic Expression Vectors of Brucella Secretory Proteins BspA and BspB and Expression in Embryonic Trophoblast Cells

WANG Shuli, WEI Shujuan, LI Menghan, ZHENG Hao, SI Lifang, LI Zhiqiang

2022, 51(8): 143-149. DOI: 10.15933/j.cnki.1004-3268.2022.08.017

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To study the role of Brucella secretory proteins BspA and BspB，two pairs of primers were designed according to BspA and BspB gene sequences of B.abortus S2308 from GenBank，and S2308 genome was used as template to amplify BspA and BspB genes by PCR.The fragments were cloned into pEGFP⁃N1 vector to obtain eukaryotic vectors pEGFP⁃BspA and pEGFP⁃BspB. After identification by enzyme digestion and sequencing analysis，LipofectamineTM 2000 was used to transfect the eukaryotic vectors into embryonic trophoblast cells（HPT⁃8）.BspAand BspB protein expressions were detected by Western blot（WB）.The secretion level of cytokines was detected by ELISA kit，and the level of cytotoxicity was detected by lactate dehydrogenase（LDH）cytotoxicity detection kit.The results showed that the full length of BspA gene was 576 bp and BspB gene was 564 bp. Eukaryotic vectors were verified by restriction enzyme EcoR Ⅰ and Xba Ⅰ digestion，and sequencing analysis showed that the homology was 100% between the sequencing information and sequence published by GenBank.WB detection results showed that the transfection groups of pEGFP⁃BspA and pEGFP⁃BspB dispalyed bands at 70 ku and 68 ku respectively，which was completely consistent with the expected results，indicating that BspA and BspB proteins could be expressed in HPT⁃8 cells.Cytokines test results showed that BspA and BspB proteins could induce HPT⁃8 cells to secrete IFN⁃γ，IL⁃2，IL⁃4 and IL⁃5.BspA and BspB proteins were non⁃toxic to HPT⁃8 cells.In summary，pEGFP⁃BspA and pEGFP⁃BspB eukaryotic expression vectors were successfully constructed，and BspA and BspB proteins could be expressed in HPT⁃8 cells and induce cytokines secretion.

Analysis of Climate Change Effects on Winter Wheat Sowing Area and Yield in Northern Henan Based on GEE

LI Feifei, TANG Jun, GAO Xianjun, YANG Yuanwei, ZHAN Yangying

2022, 51(8): 150-165. DOI: 10.15933/j.cnki.1004-3268.2022.08.018

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In order to study the impact of climate change on the sown area and yield of winter wheat in Northern Henan，the Landsat image data from 1985 to 2019 were constructed by using Google Earth Engine （GEE）cloud platform.The distribution information of winter wheat was extracted by random forest（RF）algorithm，and the changes in the sowing area of winter wheat in northern Henan were analyzed.Combined with the meteorological observation data of 9 known meteorological stations in northern Henan，spatiotemporal trends of the average temperature，precipitation，sunshine hours at the sowing date，jointing⁃heading stage，and heading⁃milk ripening stage were analyzed.The data of sowing area and yield in the study area were quantitatively analyzed.The correlation analysis，importance evaluation and sensitivity analysis of winter wheat yield to climate were carried out.The results showed that，the relative errors of the classification and extraction results of RF algorithm were all within 3.5%，the average accuracy of sample points was 94.94%，and the Kappa coefficient was between 0.88 and 0.94，reaching a high precision level.The total sowing area of winter wheat showed an increasing trend from 1985 to 2019，mainly distributed in the central and eastern regions，and the sowing frequency was higher in the central region.The temperature increasing at sowing date had a positive effect on the sowing area（P=0.013<0.05）.The importance of climate factors on the sowing area was ranked as follows：average temperature > sunshine hours > precipitation at sowing date. Winter wheat yield showed a trend of increasing in northern Henan.Precipitation at jointing⁃heading stage significantly promoted winter wheat yield（P=0.003<0.01）with a relatively higher contribution rate，while sunshine hours significantly inhibited winter wheat yield（P=0.022<0.05）.The yield of winter wheat was most sensitive to sunshine hours during heading⁃milk ripening stage. The order of importance of climate factors on winter wheat yield in northern Henan was as follows：precipitation at jointing⁃heading stage>average temperature at jointing⁃heading stage>precipitation at heading⁃milk ripening stage>sunshine hours at jointing⁃heading stage>sunshine hours at heading⁃milk ripening stage>average temperature at heading⁃milk ripening stage

Optimization of the Formula for Nutritive Cornus officinalis Jujube Slices

WANG Anjian, GAO Shuaiping, CUI Guomei, LI Shunfeng, WEI Shuxin, TIAN Guangrui, LIU Lina, XU Fangfang

2022, 51(8): 166-173. DOI: 10.15933/j.cnki.1004-3268.2022.08.019

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In order to optimize the formula of nutritive Cornus officinalis jujube slices，Cornus officinalis and jujube were used as main raw materials to prepare composite Cornus officinalis jujube slices through the steps of blending，beating，boiling，drying and slicing.With the texture and sensory score as the indexes，the optimum formula parameters of composite Cornus officinalis jujube slices were determined through single factor test and orthogonal optimization test.The optimal formula was showed as follows：jujube 100 g，Cornus officinalis 16.7 g，starch 5.0 g，citric acid 0.6 g and refined cane sugar 25 g.The prepared composite Cornus officinalis jujube slices had unique flavor，sweet and sour taste and moderate hardness，and the sensory score was 92.63，which was similar to the texture characteristics and sensory score of the commercial jujube slices.The results showed that the optimized nutritive Cornus officinalis jujube slices had a certain market acceptance.

Variation Analysis of pH Values among Different Sections of Flue⁃cured Tobacco Leaf from Guangdong

PAN Gaowei, GUO Weixiong, HAN Jiatong, HE Feng, LIANG Miao, BAI Guoqiang

2022, 51(8): 174-180. DOI: 10.15933/j.cnki.1004-3268.2022.08.020

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In order to study the variation of tobacco leaf pH values at different cutting locations of flue⁃cured tobacco in Guangdong Province，eighteen tobacco leaf samples of four flue⁃cured tobacco cultivars from three regions in Guangdong were selected.And the pH variation behavior of water extracts at ten cutting locations from leaf tip to base area was analyzed.The results showed that，the pH values of 180 leaf portions at different locations ranged from 5.31 to 5.96，and 73.3% of the samples were concentrated in the range of 5.49—5.67.There was a significant positive correlation between leaf pH value and cutting location，and the pH value from leaf tip to base showed a parabola trend of first increasing and then decreasing.The pH value of flue⁃cured tobacco in Guangdong Province showed significant or extremely significant differences among the producing areas，varieties and parts.In particular，a highly significant correlation existed between the tobacco grade and pH value. On the whole，the pH value of flue⁃cured tobacco of different grade presented the following characteristic：X2F > C3F >B2F.The cluster analysis based on tobacco leaf pH value difference indicated that the three⁃stage cutting scheme was optimal for different positions of tobacco.The leaf location of TL1—TL3 was the first section，TL4—TL7/8 was the second section，and the rest of the leaf base was the third section.

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