Journal of Henan Agricultural Sciences

Progress on Nutritional Value of Schizochytrium limacinum and Its Application in Animal Production

ZHANG Xinming, ZHAO Lu, HU Xiaowen

2021, 50(3): 1-10. DOI: 10.15933/j.cnki.1004-3268.2021.03.001

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Schizochytrium limacinum is rich in nutritional value,especially essential fatty acids such as DHA required for animal growth,so it has good application prospects in animal production.The basic nutritional value and biologically active substances of Schizochytrium limacinum were reviewed,and the influence of environmental factors on the production of Schizochytrium limacinum DHA was analyzed.The application of Schizochytrium limacinum in improving the growth performance and survival rate of animals,increasing the nutritional value of products,and promoting the fat metabolism and improving non-specific immunity of the body was reviewed.The application prospects of Schizochytrium limacinum in animal feed and additives were prospected, aiming to provide reference for the development and application of Schizochytrium limacinum.

Mechanism of Protein Kinase SAPK2 in Response to Potassium Deficiency Stress in Rice

LOU Dengji, CHEN Zhen, LI Qinyan, CHEN Qin, ZHAO Qiuhong, YANG Xiaoyan, YU Diqiu

2021, 50(3): 11-16. DOI: 10.15933/j.cnki.1004-3268.2021.03.002

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In order to reveal the function and mechanism of SAPK2(osmotic stress/ABA-activated protein kinase 2) involved in rice(Oryza sativa L.) response to K deficiency,the sapk2 mutants constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/Cas9) technology and wild type rice seeds were used to study SAPK2 expression profiles, germination, growth of seedling and the expression levels of K⁺metabolism-related genes under K deficiency condition. The results showed that the N,P and K deficiency treatments caused a significant decrease in the expression level of SAPK2 in rice;compared with the wild type, the germination of the sapk2 mutants was significantly delayed, the germination rate was significantly reduced,and the seedling growth was severely inhibited under the K deficiency condition; compared with wild-type,the expression levels of K⁺ metabolism-related genes such as OsKAT1(potassium channel gene1),OsKAT2(potassium channel gene 2),OsAKT1(shaker K⁺channel1) and OsHAK1 (high-affinity potassium transporter1) in sapk2 mutants were significantly downregulated.These results indicated that SAPK2 played an important positive regulatory role in the process of seed germination and seedling vegetative growth of rice under K deficiency environment,and the mechanism may be that SAPK2 directly or indirectly regulated the expression level of K⁺ metabolismrelated genes to participate in the K deficiency response.

Effects of Combined Application of Nitrogen and Potassium on Lodging Resistance of Rice in Cold Region under Condition of Returning Straw to Field

ZHANG Xuesong, ZHANG Peng, WANG Xiuzhi, CHEN Rongfa, WANG Wenyu, WAN Siyu, SONG Weimin, ZHANG Gongliang, LI Yi, QIAN Yongde

2021, 50(3): 17-24. DOI: 10.15933/j.cnki.1004-3268.2021.03.003

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In order to solve the problem of stem lodging caused by returning straw to the field and improper fertilizer application in the northern cold regions,the rice variety Kenjing No.8 was used as the material,a two-factor random block design was adopted,the effect of combined application of different nitrogen and potassium( N1:62. 592 kg/ha,N2:125.184 kg/ha,N3:187.776 kg/ha;K1:45 kg/ha,K2:90 kg/ha,K3:135 kg/ha)on lodging resistance of rice was studied under the condition of returning straw to the field in rice main production region in cold regions.The results showed that under the condition of full amount of straw returning to the field,nitrogen fertilizer had significant or extremely significant effects on the height of gravity center, ear length, length of second and third internodes,diameter of third and fourth internodes,thickness of stem wall,breaking resistance,anti-breaking moment and lodging index; potassium fertilizer had significant or extremely significant effects on height of gravity center,length of third and forth internodes,diameter of third internode,thickness of stem wall,breaking resistance and anti-breaking moment,bending moment of second and third intemodes and anti-breaking moment;the interaction of nitrogen and potassium mainly affected the length of internodes, height of gravity center,breaking resistance and anti-breaking moment.Under low and middle nitrogen(N1,N2) conditions,additional application of potassium fertilizer was conducive to improve the breaking resistance,reduce lodging index; under high nitrogen(N3) condition,excessive application of potassium fertilizer reduced the plant’ s breaking resistance,and increased lodging index.Among the different combination treatments,the lodging index of N2K3 treatment was the smallest,followed by N1K3 treatment.Under the condition of full amount of straw returning to the field in the cold area,the optimal application plan resistant to lodging of nitrogen and potassium for Kenjing No.8 was N2K3 treatment (N:125.184 kg/ha+K₂O 135 kg/ha),and the ratio of nitrogen and potassium was 0.927∶1.

Mechanism of High Nitrogen Utilization Efficiency of Indica Rice Yliangyou 886

QI Xuejiao, XIE Qiankun, XIE Yuxin, LIANG Xiaoya, LI Junzhou, SUN Huwei

2021, 50(3): 25-32. DOI: 10.15933/j.cnki.1004-3268.2021.03.004

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To determine the mechanism of high nitrogen utilization efficiency of indica rice Yliangyou 886,the biomass, total nitrogen ( N) content, root development, nitric oxide(NO) levels of root and expression levels of N transport genes in root were analyzed in Nipponbare (japonica) and Y liangyou 886(indica) under two N concentrations(normal N concentration and low N concentration) by hydroponics and soil culture experiments.The results showed that the biomass,total N content and nitric oxide level of root were increased significantly in Yliangyou 886 compared with those in Nipponbare under the two N concentrations.However,the lateral root density of the two cultivars did not change significantly under two N concentrations. qRT-PCR results showed that the expression levels of several N transport genes in the roots were significantly up-regulated in Yliangyou 886 compared with Nipponbare under two N concentrations.To sum up,root of Yliangyou 886 has a strong ability to synthesize nitric oxide,thereby promoting the expression of several N transport genes and improving the N use efficiency.

Effect of Chemical Regulator on Photosynthetic Characteristics,Leaf Senescence and Yield of Spring Maize under Density-Increasing Condition

LI Chao, WANG Ji, ZHU Min, LI Fenghai

2021, 50(3): 33-41. DOI: 10.15933/j.cnki.1004-3268.2021.03.005

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To explore the regulation mechanism of chemical regulator on photosynthetic characteristics and leaf senescence of spring maize under density-increasing condition,a field experiment was conducted using Zhengdan 958 (ZD958,compact type) and Liangyu 99 (LY99,semi-compact type) with chemical regulator( Gaoyubao, active ingredient was 40% ethephon) under different planting densities [52 500(D1),60 000 (D2),67 500 ( D3),75 000 (D4)plants/ha],and the effect of chemical regulator on photosynthetic characteristics, leaf senescence and yield of spring maize was studied under densityincreasing
condition.The results showed that spraying chemical regulator significantly reduced plant height and ear height of spring maize. The SPAD value of ear leaf also increased after spraying chemical regulator at silking and filling stages, which delayed leaf senescence. Spraying chemical regulator improved the photosynthetic performance of ear leaf, increased photosynthetic rate, transpiration rate,stomatal conductance, activities of antioxidant enzymes (peroxidase, superoxide dismutase) and key enzymes(nitrate reductase, glutamine synthetase)related with nitrogen metabolism, promoted nitrogen assimilation and metabolism of leaf,and delayed leaf senescence at late stage. For CK,the yields of ZD958 and LY99 increased first and then decreased with the increase of planting density;the yield of ZD958 was the highest under D3 density,and which of LY99 was the highest under D2 density. After spraying the chemical regulator,overall the yields of both two spring maize hybrids were the highest under D2 density.Thus,reasonable planting density combined with spraying chemical regulator could be an important technique for high-yield cultivation of spring maize in Northeast China.

Effect of Late Spring Coldness on Quality of High-Quality Strong Gluten Wheat Zhengmai 366

QIN Maomao, LIU Yanxi, ZHANG Kun, ZHAO Mengqi, ZHOU Zhengfu, WU Zhengqing, CHEN Feng, LEI Zhensheng

2021, 50(3): 42-48. DOI: 10.15933/j.cnki.1004-3268.2021.03.006

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In order to understand the effect of late spring coldness on processing and nutrient quality of high-quality strong gluten wheat Zhengmai 366,quality indexes of Zhengmai 366 with different degree of freezing damage were analyzed.The results showed that overall the crude protein content, wet gluten content and dough water absorption increased with the deepening of freezing damage; however,the volume weight and dough stability time decreased with the deepening of freezing damage,and the processing quality of flour decreased.For nutritional quality, the flour total protein content increased with the deepening of freezing damage;compared with the control,the amylose contents of Ⅲ,Ⅴ types of wheat significantly decreased,and amylopectin contents of Ⅲ,Ⅴ types of wheat significantly increased; the total starch contents of Ⅲ,Ⅳ types of wheat and ratios of amylose to amylopectin of Ⅲ,Ⅴ types of wheat significantly decreased compared with the control.The starch pasting properties results showed that the peak viscosity,trough viscosity,breakdown value,final viscosity and setback value decreased with the deepening of freezing damage except for Ⅳ type,Ⅰ and Ⅳ types of wheat had greater influence on the starch pasting properties,and the gelatinization quality decreased under late spring coldness.

Investigation and Utilization of Selenium-Enriched Medicinal Plants in Enshi,Hubei Province

ZHOU Wuxian, ZHANG Yajuan, ZHANG Meide

2021, 50(3): 49-58. DOI: 10.15933/j.cnki.1004-3268.2021.03.007

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In order to utilize the selenium ( Se)-enriched soils and medicinal plants and promote the development of Se-enriched medicinal products in Enshi Tujia and Miao Autonomous Prefecture(hereafter it will be abbreviated as Enshi Autonomous Prefecture).An investigation was conducted to test the Se concentration of soils(187 samples) and medicinal plants (243 samples) from twelve sampling sites in Enshi Autonomous Prefecture, and an experiment was conducted to learn the effects of foliar Se application on the growth of three medicinal plants(Gynostemma pentaphyllum,Atractylodes macrocephala and Codonopsis tangshen)from 2016—2018.The results showed that the soil Se content of 83.3% of the twelve investigated sampling sites met the standard of high level (Se>0.4 mg/kg),and the Se content of soil from Yutangba (Xintang Township,Enshi) reached extremely high level (Se>3 mg/kg).The plant Se content of 23.8% of the 168 investigated medicinal plants met the standard of Se-enriched agricultural products (Se≥0.2 mg/kg),among which Cardamine violifolia was superior in accumulation of Se with Se content of 311.00 mg/ kg and Se accumulation coefficient (SAC) of 76.60.There were differences in SACs among different species of medicinal plants which were cultivated in the same sampling site,and the same medicinal plants cultivated in different sampling sites also had different SACs.It was an effective way to produce Se-enriched medicinal products through cultivating medicinal plants in typical seleniferous region or spraying Se on plant leaves in Se deficient region. In conclusion, there were numerous Seenriched soils and medicinal plants in Enshi Autonomous Prefecture,and cultivating medicinal plants in typical seleniferous region or spraying Se on plant leaves in Se deficient region could be beneficial for the development of safe Se-enriched medicinal products in Enshi Autonomous Prefecture.

Effect of Seed Dressing with Ammonium Molybdate on Development and Nitrogen Accumulation of Peanut in Poor Dry Land

ZHANG Jun, HAO Xi, LIU Juan, ZANG Xiuwang, ZHANG Man, TANG Fengshou, DONG Wenzhao

2021, 50(3): 59-66. DOI: 10.15933/j.cnki.1004-3268.2021.03.008

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In order to improve the yield and quality of peanut in dry and thin land and alleviate the lack of molybdenum in peanut planting soil,pot experiments were conducted to set four ammonium molybdate seed dressing amounts of 0(CK),0.5,1.0 and 1.5 g/kg to analyze the effects of ammonium molybdate seed dressing on peanut growth and nitrogen accumulation, and to explore the optimal ammonium molybdate seed dressing amount.The results showed that the seed dressing with ammonium molybdate could promote the vegetative growth and reproductive growth of peanut, improve the leaf area and photosynthetic rate,increase the nitrogen content of organ and yield,and improve the quality of peanut.Among them,application of 1.0—1.5 g/kg ammonium molybdate had the best effect.Compared with the control group, application of 1.5 g/kg ammonium molybdate could increase the flower number, fruit number,pod number per plant and pod volume by 48.43,19.46%,26.49% and 16.39%,improve the leaf area,leaf net photosynthetic rate and productivity per plant by 25.20%,51.08% and 32.74% respectively.Tests had shown that the application of ammonium molybdate could significantly increase peanut vegetative and reproductive growth, promote nitrogen accumulation in various organs, increase peanut yield,and improve peanut quality.

Phosphorus Pool and Agronomic Effects of Phosphorus Fertilizer Reduction in Rice-Wheat Rotation Field along the Yellow River and Huai River of China

ZHANG Yangyang, ZHANG Shuli, XIE Yingxin, KANG Guozhang, CHEN Bo, MA Dongyun, WANG Chenyang, GUO Tiancai

2021, 50(3): 67-73. DOI: 10.15933/j.cnki.1004-3268.2021.03.009

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In order to explore soil phosphorus(P)deliverbility in rice-wheat rotation area along Yellow and Huai Valleys in Henan Province and suitable P reduction regimes, a field fixed experiment was carried out to study effect of P reduction by investigating soil P pool status and analyzing the soil available P and grain yield in three typical soils (Yuanyang,Fengqiu and Xinyang of Henan Province) in rice wheat rotation system along the Yellow River and Huai River of China. The experiment had four fertilization regimes,P fertilization during the rice and wheat season(WRP,current farming practice),Pfertilization only during the wheat season(WP),P fertilization only during the rice season(RP),and no P fertilization during either season (NP ).The results showed that the available P content presently reached 15. 10 mg/ kg in topsoil in Henan Province, which was 9.40 mg/kg (the increase range of 165.0%)and 2.37 mg/kg(the increase range of 18.6%)higher than those of the second soil survey and the national soil testing formula survey,respectively.Moreover,the available P content in topsoil had significant differences among three experimental sites,the highest was in Yuanyang,followed by Fengqiu,and the lowest was in Xinyang. And the available P content in topsoil at rice maturity was significantly higher than that at wheat maturity,but significant differences among RP,WP and WRP treatments at rice maturity were not found. In addition,no significant differences were also found in the annual grain yield of rice and wheat among three P fertilization treatments(WRP,WP and RP),especially between WRP and WP treatments.In a conclusion,based on current status of the soil P in the typical farmland of rice-wheat rotation along the Yellow River and Huai River of China,the soil P content has basically met the needs of crop growth.Therefore,the regime of P fertilizer application only in wheat season(WP)not only ensures adequate soil P supply to maintain grain yield of rice and wheat,but also has the potential to save P resources and reduce the risk of P pollution in water body.

Effect of Chemical Fertilizer Substitution by Organic Fertilizer and Water Saving Irrigation on Runoff Loss of Nitrogen and Phosphorus in Vegetable Field

LÜ Hongwei, LUO Xiaosheng, KOU Changlin, WANG Hongke, CHEN Songqin, LÜ Jing

2021, 50(3): 74-80. DOI: 10.15933/j.cnki.1004-3268.2021.03.010

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In order to clarify the characteristics of nitrogen and phosphorus runoff loss in vegetable fields in Xinye County, Henan Province,the effects of organic fertilizer instead of chemical fertilizer and watersaving irrigation on nitrogen and phosphorus runoff in open vegetable field (two seasons in one year,cabbage-eggplant rotation)were studied by field location experiment from 2016 to 2018. Three treatments were set up,namely conventional fertilization (for short,conventional),organic fertilizer replacing 27.5%chemical fertilizer (organic fertilizer replacing),organic fertilizer replacing 27. 5% chemical fertilizer+30% water-saving irrigation (organic fertilizer replacing +water-saving irrigation).The results showed that there were 10 runoff occurences in total (1 in 2016, 6 in 2017 and 3 in 2018),and the annual runoff production was 586.3—3497.3 m³/ha during the 3 years experiment. During the 3-year experiment,the average contents of nitrate nitrogen in runoff water were 9.55,8.14,7.33 mg/L for conventional,organic fertilizer replacing,and organic fertilizer replacing +water-saving irrigation treatments, respectively.The contents of soluble total phosphorus were 0.96,0.85,0.65 mg/L,respectively.The average runoff losses of nitrate nitrogen were 4.87,4.07,3.58 kg/ha,respectively.The runoff loss of soluble total phosphorus were 0.49,0.43,0.32 kg/ha.Compared with the conventional treatment, organic fertilizer replacing treatment reduced nitrate nitorgen and soluble total phosphorus runoff by 16.43% and 12.24% on average in 3 years.The organic fertilizer replacing treatment and water-saving irrigation treatment reduced the runoff of nitrate nitrogen and soluble total phosphorus by 26.48% and 34.69% on average in 3 years.There was no significant difference in vegetable yield among different treatments.The results indicated that organic fertilizers instead of partial chemical fertilizers and proper water-saving irrigation could reduce nitrogen and phosphorus loss in vegetable fields on the basis of maintaining vegetable yield in the open vegetable fields in southwest Henan Province.

Experiment and Simulation of Nitrate and Ammonium Nitrogen Transport under Wide Ridge and Furrow Irrigation with Different Fertilizer Content

WANG Shunsheng, YAN Yongfang, CHEN Chunlai, KOU Jianhui, HAO Li

2021, 50(3): 81-90. DOI: 10.15933/j.cnki.1004-3268.2021.03.011

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In order to explore the distribution of nitrate and ammonium nitrogen in broad ridge and furrow irrigation and the effect of different fertilizer solution contents on nitrogen content,the nitrogen transport under three different fertilizer solution contents (300,600 and 900 mg/L) was simulated by using the software of HYDRUS-2D,and the distribution of nitrate and ammonium nitrogen under different fertilizer solution contents was analyzed. The results showed that nitrate nitrogen and ammonium nitrogen had the same trend in the infiltration stage under different fertilizer liquid contents.With the increase of fertilizer liquid from 300 mg/L to 900 mg/L,the distribution range of nitrate nitrogen and ammonium nitrogen increased with the process of fertilizer liquid infiltration,and the nitrate nitrogen and ammonium nitrogen content decreased gradually from near to far away from the irrigation ditch;The nitrate nitrogen content tended to be 29.3 mg/kg at the distance of 50—60 cm from the center point D of irrigation ditch,and the ammonium nitrogen content tended to be 7.7 mg/kg at the distance of 20—40 cm from the center point D of irrigation ditch,which indicated that ammonium nitrogen had a faster rate of deceleration with the increase of distance than nitrate nitrogen in the process of migration,and the effect of adsorption on ammonium nitrogen content was more significant; the HYDRUS-2D software had good results in the simulation of nitrogen migration and distribution,which provided a scientific basis for the study of nitrogen migration and reasonable fertilization in wide ridge and furrow irrigation.

Control Effect of Burkholderia pyrrocinia YZU-S377 on Cotton Verticillium Wilt and Its Growth Promotion Function

LIU Lu, WAN Weijie, ZHENG Tongwen, LONG Xinyu, SUN Zhengxiang, ZHOU Yi

2021, 50(3): 91-96. DOI: 10.15933/j.cnki.1004-3268.2021.03.012

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Burkholderia pyrrocinia YZU-S377(S377) is a rice rhizosphere bacterium obtained in the early stage,which has broad-spectrum inhibitory effect on plant pathogens.To verify the biocontrol potential of S377 against Verticillium dahliae,the control effect of S377 on cotton Verticillium wilt,the colonization dynamics and growth promoting effect of S377 on cotton were tested in pot. The results showed that the relative inhibition rate of the strain to V.dahliae was 89.22%,and the control effect of pot test was 62.53%.After treated with S377 sterile filtrate the germination rate of cotton increased by 22.22 percentage points,the root length of cotton seedlings increased by 27.78 mm,and the fresh weight of aboveground part increased by 0.73 g.The results of crystal violet staining showed that strain S377 had the ability to form biofilm.The biofilm formation intensity of strain S377 at OD₆₀₀=0.15 and 0.25 was significantly more obvious than that under other concentrations.The colonization experiment showed that S377 could colonize cotton roots for 21 days,and the number increased first and then decreased. The
results indicated that S377 was a potential biocontrol strain against cotton Verticillium wilt.

Identification and Analysis of Pathogen of Kiwifruit Rot in Guizhou Province

SU Wenwen, WU Di, HAN Zhencheng, LI Liangliang, LI Weijie, REN Chunguang

2021, 50(3): 97-102. DOI: 10.15933/j.cnki.1004-3268.2021.03.013

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In order to clarify the types of the pathogens of fruit rot disease of main kiwifruit varieties in Guizhou Province,and provide theoretical basis for the prevention and control of field fruit diseases,the diseased fruits of Hongyang and Guichang kiwifruit collected in Shuicheng County and Xiuwen County of Guizhou Province from 2018 to 2019 were used as experimental materials.The pathogens were identified through pathogen isolation,morphological observation,molecular detection and pathogenicity detection.The phylogenetic analysis of pathogen species based on their sequences were carried out.The results showed that four kinds of pathogens including Phomopsis cauloides, Fusarium, Alternaria alternata,Plectosphaerella cucumerina were isolated, and their detection rates were 10%, 60%, 20%, 10%respectively. The fruit disease of kiwifruit caused by the infection of Plectosphaerella cucumerina was reported for the first time in China.

Study on the Preparation Methods of Zoospores of Phytophthora infestans

YANG Zhuqiang, WANG Hongyang, TANG Wei, LI Canhui, LIU Jing

2021, 50(3): 103-108. DOI: 10.15933/j.cnki.1004-3268.2021.03.014

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Phytophthora infestans is the pathogen of potato late blight, which poses a serious threat to potato production.In order to explore the optimal preparation conditions for the zoospores of P.infestans,after cultivation for 7—12 days,Petri’s solution was added to cultures,and sporangia was collected and counted.Five different low-temperature induction methods were used to prepare zoospores,and the effects of different culture time and different preparation methods on zoospore yield were studied.The lowtemperature induction methods were as follows:M1,the sample was stood at 10℃ for 30 min and then at 18℃ for 30 min;M2,the sample was stood at 4℃ for 30 min and then at 10℃ for 90 min;M3,the sample was left at 4℃ for 30 min and then at 10℃ for 60 min;M4,the sample was stood at 4℃ for 3 h;M5,the sample was stood at 4℃ for 2 h and then at 18℃ for 1 h.The results showed that with the extension of the cultivation time, the yield of sporangia increased rapidly first and then slowed down,finally basically stopped. As the cultivation time of sporangia increased,the zoospore yields obtained by different preparation methods increased first and then decreased.The yield of zoospores was the highest when sporangia were harvested from 9 day culture and there was no significant difference among different preparation methods.In summary,to quickly obtain zoospores in large quantities,the sporangia of P.infestans cultured for 9 days was stood at 10℃ for 30 min and then at 18℃ for 30 min.Or regardless of the preparation time,the 9-day-old sporangia was stood at 4℃ for 2 h and then at 18℃ for 1 h to obtain the largest number of zoospores.The yield of sporangia is not completely positively correlated with the yield of zoospores.

Study on the Inhibition Mechanism of Fermentation Material of Pleurotus ostreatus on Mold

CUI Xiao, LIU Qin, DING Yatong, KANG Yuanchun, HU Sujuan, SONG Kaibo, ZHANG Yuting, KONG Weili

2021, 50(3): 109-116. DOI: 10.15933/j.cnki.1004-3268.2021.03.015

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After fermentation of culture meterial,Pleurotus ostreatus was cultivated by open inoculation method,which could effectively inhibit the growth of mold and improve the survival rate of the bag,but its antibacterial mechanism was still unclear.In this study,corn cob was used as the main material to study the effect of its extract on mycelium growth,mycelium biomass,spore germination rate,cell morphology,the enzyme activity of succinodehydrogenase,adenosine triphosphatase,and enzyme content of alkaline phosphatase of Trichoderma atroviride and Penicillium glaucum.The results indicated that during the fermentation period,the inhibitory effect of culture material on T.atroviride and P.glaucum increased gradually with the extension of fermentation time,and the highest inhibition rate of the extract on the two kinds of mycelium was 100.0% and 97.9%, respectively at the T4 stage.With the increase of the concentration of the extract liquid,the inhibition rate increased,the mycelium biomass decreased,and the inhibition rate of 100% treatment was 81.1%、75.9% and the hyphae biomass of T.atroviride and P.glaucum reduced by 66.2% and 64.6% respectively.Microscopic findings showed that after the culture of 80% treatment,T.atroviride mycelium cell wall and cell membrane were broken,vacuole and mitochondrial membrane were broken,ribosome distribution was uneven,cell content decreased,and the normal growth was hindered;and the edge of P.glaucum mycelium cell wall was rough and the diaphragm was broken.With the increase of extract concentration,the AKP content of T.atroviride and P.glaucum increased,which was 333.3% and 560.0% higher than that of the control.The activity of SDH enzyme related to respiration decreased to the lowest in 48 hours,which was 8.1 and 10.3 U/mg respectively.Both of the activity of ATP was lower than that of control group,which was 51.3% and 55.8% lower than that of the control group.The study showed that after fermentation of P.ostreatus culture material,the extract of P.ostreatus destroyed the cell wall and cell membrane of two kinds of mycelium,which resulted in the outflow of intracellular lysates and reduced mycelium metabolism and respiratory intensity,so the mycelium growth and spore germination of Trichoderma atroviride and Penicillium glaucum were inhibited.

Evaluation of Drought Resistance of Different Walnut Varieties Based on Leaves Structure Analysis

YUN Linying, LIU Kai, LI Linqing, ZHAO Shugang, WANG Hongxia

2021, 50(3): 117-124. DOI: 10.15933/j.cnki.1004-3268.2021.03.016

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The leaf structure(19 indexes included leaf thickness,the upper epidermis thickness and the lower epidermis thickness,palisade tissue thickness,spongy tissue thickness,main vein thickness,vascular bundle thickness,phloem thickness,stomatal size,stomata density,and so on) of 5 walnut varieties[Black walnut(Juglans regia),Qingxiang(J.regia),Liaoning 1(J.regia),Bread crossing walnut(J.major×J.regia),YZ2 (J.regia)] were studied by paraffin sectioning and glue imprinting. Based on principal component analysis to screen drought indexes, drought resistance of different walnut varieties was evaluated, which would provide a theoretical basis for drought-resistant breeding of varieties and rootstock.The results showed that there were some differences among the indexes except for the upper epidermis thickness in different walnut varieties,and the coefficient of variation was between 9.07% and 48.10%.The leaf thickness, palisade tissue thickness, sponge tissue thickness, main vein thickness,vascular bundle thickness and xylem thickness of Black walnut and Breedcrossing walnut were significantly shorter than those of Qingxiang cultivars;Leaves thickness and palisade tissue thickness of Liaoning 1 were significantly larger than those of the other two Black walnut cultivars;Vein thickness,vascular bundle thickness and stoma density of Qingxiang were significantly larger than those of the other varieties.Based on principal component analysis, leaf thickness, vascular bundle thickness, stomatal length,the lower phloem thickness and the lower parenchyma thickness were screened as indexes for comprehensive evaluation of drought resistance. Drought resistance of five walnut varieties was evaluated by membership function analysis.The order of drought resistance was: Black walnut> Qingxiang>Liaoning1>Breed-crossing walnut>YZ2.

Study on Application of Honeybee Activity Enhancer on Sweet Cherry

WANG Chengxiang, HU Haiyan, ZHANG Ruihua, LIU Ying, LIU Zhenlong

2021, 50(3): 125-129. DOI: 10.15933/j.cnki.1004-3268.2021.03.017

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In order to improve the yield and quality of sweet cherry, the effects of honeybee activity enhancer on honeybee activity,fruit setting rate,yield and commodity rate of sweet cherry were studied with sweet cherry variety Meizao as the experimental material.The results showed that the number of flower-visiting Osmia muricata was increased obviously,and the number of flower-visiting Osmia muricata per unit time on fruit trees in the experimental area(hanging Baolirui) which were 15 meters away from the hive,was the highest with 4.22 heads/(plant·min),2.14 heads/(plant·min) more than the control(no Baolirui),and 1.77 heads/(plant·min)more than the control on the 6th day of observation.The fruit setting rate and yield of cherry were also significantly affected by Baolirui. The fruit setting rate in the test area 15 meters away from the hive increased by 10.77 percentage points,the yield per plant increased by 3.52 kg and equivalent yield increased by 2 640 kg/ha.There was no significant effect on the commercial rate of cherry fruit,but the fruit quality of treated area was slightly better than that of control area.To sum up,we can use honey bee activity enhancer to improve the yield and quality of cherry in the future production, and then promote the development of cherry industry.

Effects of Foliar Application of Organic Selenium on Total Flavonoids Content in Ginkgo biloba Leaves

XIAO Xian, LI Li, LI Linling, ZHA Sanxing, YUAN Honghui, DENG Kun, CHENG Shuiyuan, CHENG Hua

2021, 50(3): 130-141. DOI: 10.15933/j.cnki.1004-3268.2021.03.018

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In order to study the effect of selenium on the content of total flavonoids in Ginkgo biloba leaves,the four-year-old field ginkgo seedlings were sprayed with seven kinds of treatments once or twice on leaves,the sprayed materials were no fertilizer (CK), 0.835 g/L organic selenium (T1), 1 g/L chlorocholine chloride(T2),2 g/L chlorocholine chloride(T3),1 g/L chlorocholine chloride and 0.835 g/L organic selenium(T4),2 g/L chlorocholine chloride and 0.835 g/L organic selenium(T5),as well as 3.335 g/L potassium dihydrogen phosphate and 0.835 g/L organic selenium(T6) respectively.Both the changes of the content of total flavonoids of Ginkgo biloba leaves and the expression level of genes related to the synthesis of flavonoids were detected and analyzed.The results showed that T1 treatment had the best increase effect on total flavonoids content.The Ginkgo biloba leaves(treated by organic selenium once) sampled on June 30th,August 1st,August 15th,August 31st and September 15th increased the contents of total flavonoids by 36.71%,4.93%,19.28%,21.91% and 17.68% compared with CK,respectively.In addition,those treated by organic selenium twice and sampled on August 1st, August 15th,August 31st and September 15th improved the contents of total flavonoids by 44.02%,39.86%,6.29% and 27.35%, respectively.The results of expression level showed that PAL(Phenylalanine ammonia lyase),C4H(Cinnamic acid 4-hydroxylase),4CL(4-coumaric acid coenzyme A ligase),CHS(Chalcone synthase),CHI(Chalcone isomerase),IFS(Isoflavone synthase),FS(Flavonoid synthase),F3H(Flavanone 3-hydroxylase ),FLS(Flavonol synthase ),FL3H(Naringenin 2-oxoglutarate 3-dioxygenase),OMT(Quercetin O-methyltransferase) and F3′5′H (Flavonoid 3′,5′ hydroxylase) genes were up-regulated under T1 treatment.The results also indicated that the expression level peaks of PAL,C4H,OMT and CHI genes were earlier than CK,and 4CL,CHS,FLS genes were kept higher expression level than CK.Besides,organic selenium combined with chlorocholine chloride could effectively promote the early expression of key genes related to the synthesis of flavonoids in Ginkgo leaves,which maintaed a high level,then improving the contents of total flavonoids.

Isolation,Identification and Nuclease Activity of Extracellular Secretory Protein of Klebsiella pneumoniae in Mink

JIA Yanyan, LI Qi, WANG Xiaoli, NIU Junhui, MAO Jing, LIAO Chengshui, ZHANG Haokai, ZHANG Mingliang

2021, 50(3): 142-147. DOI: 10.15933/j.cnki.1004-3268.2021.03.019

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In order to identify the pathogens causing mink pneumonia,a suspected strain of Klebsiella pneumoniae was isolated from clinical death cases.The nuclease activity of extracellular protein of Klebsiella pneumoniae was detected by agarose gel electrophoresis, and the effects of temperature, pH value and metal ions on the activity of nuclease were investigated. The results showed that Klebsiella pneumoniae was identified by microscopic examination and PCR.The exocrine protein of Klebsiella pneumoniae showed the activity of nuclease for degradation of λDNA.The optimal temperature range was 30—37 ℃ ,and the optimum pH value was 7.0.The activity of nuclease was weak under high temperature and alkaline conditions.Univalent metal ions (Na⁺and K⁺) and some divalent metal ions (Ca²⁺,Ba²⁺,Ni²⁺,Zn²⁺and Cu²⁺) had no significant effect on the nuclease activity of extracellular secretory protein of Klebsiella pneumoniae.The 5.00—10.00 mmol/L Co²⁺and Fe³⁺and 10. 00 mmol/L Mg²⁺and Mn²⁺could promote the activity of extracellular nuclease cleavaging λDNA.In this study,a Klebsiella pneumoniae strain was isolated from clinical mink pneumonia,and its exocrine protein had nuclease activity.

Effects of Lactobacillus plantarum LR-39 on Growth and Intestinal Microbiota Regulation of Dogs

CHEN Zedong, ZHU Jianping, GUO Shuyu, LU Wei, LIU Lei

2021, 50(3): 148-156. DOI: 10.15933/j.cnki.1004-3268.2021.03.020

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To evaluate the growth-promoting and intestinal flora regulation effects of Lactobacillus plantarum LR-39 isolated from fermented pickles,ten 3.5-month-old beagle dogs were divided into control group and experimental group. The control group was fed with basic dog food,while the experimental group was daily given LR-39 with the basic dog food for 28 days.The average daily gain,feed weight ratio,nutrient digestibility and other indicators were measured at the end of the test. Based on highthroughput sequencing,the relative abundance of intestinal flora at the phylum level and the differences in the characteristic microbes of each group of intestinal flora were analyzed. PICRUSt was performed to further analyze the expression difference of microbial functional genes in the intestinal tract.The whole genome sequencing of LR-39 and subsequent COG and CAZy annotation analysis were performed.The results of the growth-promoting test showed that the average daily gain of the experimental group was significantly(P<0. 01) higher than that of the control group;the feed weight ratio of the experimental group was significantly(P<0.05) lower than that of the control group.The digestibility of crude protein,crude fat and calcium in the experimental group was significantly(P<0.05) higher than those of the control group.High-throughput sequencing results showed that at the phylum level,the relative abundance of Fusobacteria in the experimental group was significantly(P<0.05) higher than that in the control group,while the relative abundance of Actinobacteria was significantly(P<0.05) lower than that in the control group.LDA effect size(LEfSe) analysis showed that the characteristic bacterial generus of the intestinal flora in the experimental group was Fusobacterium,while the characteristic bacterial genera of the control group were Bulleidia, Catenibacterium,Eubacterium,Blautia,and Faecalibacterium.At the species level,the relative abundance of Lactobacillus helveticus in the experimental group was significantly(P<0.01)increased.Intestinal functional gene difference analysis showed that the relative expression abundance of intracellular transport and extracellular structure genes in the experimental group was significantly(P<0.05) higher than that in the control group,and the relative expression abundance of secondary metabolite gene was significantly(P<0.05) lower than that in the control group.Whole genome sequencing showed that the strain had a full length of 247 554 bp, no plasmid, with GC content of 44.26%.The abundance of genes related to carbohydrate metabolism was the highest,followed by amino acid metabolism related genes,the abundance of genes related to lipid metabolism was lower,and the genes related to antagonistic mechanism had a certain proportion.The enzyme system mainly included glycoside hydrolases, glycosyltransferases, carbohydrate esterases, carbohydrate binding molecules and auxiliary enzymes.In summary,feeding LR-39 can promote the growth of dogs,change the structure of the intestinal flora,regulate the expression of functional genes in intestinal flora,and it has the potential to develop into probiotic products.

Prokaryotic Expression and Antigenicity Analysis of Outer Membrane Protein TolC of Fish Pathogenic Aeromonas hydrophila

RONG Na, JIAN Sijie, SUN Wei, KANG Chao, CHEN Chen, LIU Xiang

2021, 50(3): 157-164. DOI: 10.15933/j.cnki.1004-3268.2021.03.021

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The aim was to explore the immunogenicity and antigenicity of outer membrane protein TolC of fish pathogenic Aeromonas hydrophila(A.hydrophila),evaluate its immunological function,and provide a theoretical basis for the development of Aeromonas hydrophila subunit vaccine. Bioinformatics method was utilized to analyze the physicochemical properties of TolC protein and analyze its homology and evolutionary relationship among bacteria. The molecular cloning method was used to develop a recombinant strain expressing TolC protein, which was purified by inclusion body washing and SDSPAGE,and was used to immunize mice to prepare TolC antiserum.Western blot was used to detect the specificity and titer of the antiserum, and enzyme-linked immunosorbent assay (ELISA) was used to simulate the recognition effect between TolC antiserum and different aquatic pathogens to explore the antigenicity of TolC protein.The results showed that TolC was a porin protein,which had high conservative evolution among Aeromonas.The TolC protein was expressed and purified successfully,and the optimal expression condition for TolC protein was strain OD₆₀₀ of 1.0, final isopropyl-β-d-thiogalactoside(IPTG)concentration of 0.1 mmol/L,inducing temperature of 37℃ and inducing time of 8 hours.Western blot and ELISA analysis showed that the antiserum of TolC held high specificity and titer(1∶3 200),and the TolC antiserum could recognize with Aeromonas hydrophila, Pseudomonas fluorescens and Vibrio alginolyticus.In summary,the outer membrane protein TolC of A.hydrophila had high immunogenicity and antigenicity,and might provide cross immune protection for different aquatic pathogens.

Preparation and Characterization of Nanocellulose from Tobacco Straw

ZHENG Jianqiang, LIU Bin, WANG Xiaofei, JIA Xuewei, XU Chunping

2021, 50(3): 165-173. DOI: 10.15933/j.cnki.1004-3268.2021.03.022

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In order to effectively utilize agricultural waste tobacco straw (TS), TS was used as raw material,cellulose nanocrystal ( CNC) was prepared by ultrasonic assisted sulfuric acid method.The products were observed by optical microscope, scanning electron micrograph, FT-IR spectra, X-ray diffraction (XRD),thermogravimetric (TG) and rheological analysis.The results showed that the yield of CNC was 22.4% when the ratio of TS to 64% sulfuric acid was 1∶15 (g∶mL),and the reaction time was 60 min at 45℃ .The prepared CNC was short rod-like structure with a diameter of 10—20 nm; XRD analysis showed that it retained natural cellulose Ⅰ structure with a relative crystallinity of 58.5%; CNC had better thermal stability, and the initial pyrolysis temperature was 252.36℃.The steady-state rheological test showed that the viscosity of TS-CNC suspension increased with the increase of solid content,and the shear thinning behavior strengthened with the increase of the viscosity.The dynamic strain test showed that CNC suspension solution exhibited obvious gel behavior when the solid content was1.00%.

Research on Identification Accurate Rate of Soybean Leaf Diseases Based on UAV Image Processing

TAN Qinhong

2021, 50(3): 174-180. DOI: 10.15933/j.cnki.1004-3268.2021.03.023

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In order to balance the contradiction between unmanned aerial vehicle(UAV) recognition coverage and recognition accuracy, and realize the identification accurate rate of large-area soybean diseases,SLIC super-pixel method was used to process soybean leaf images acquired by UAV at 1 m、2 m、4 m、8 m、16 m.After extracting the color,gradient,texture and shape characteristics of leaves,sequential minimal optimization,J48 decision tree,k-nearest neighbors and random forest algorithm were used based on the feature vectors of these attributes to identify soybean leaf diseases,and the accuracy of different algorithms for disease identification was obtained.The influence of input parameters on disease identification was analyzed by comparing the accuracy of disease recognition with feature fusion and single feature.The results showed that the accurate rates of the four algorithms were more than 90% when the UAV image shooting height was 1 m and 2 m,and the SMO algorithm and random forest algorithm had higher identification accurate rate.The identification accurate rate of soybean disease with fusion feature vector as input parameter was higher than that of single feature vector. Color was the attribute that could best reflect the actual situation of leaf disease.The optimal shooting height of UAV was 1—2 m.

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