Characterization of mulberry leaf instant tea and evaluation of its hypolipidemia effect via regulation of intestinal microbiota

The purpose of this study was to characterize mulberry leaf instant tea(MLIT)powder prepared from the'Longsang No.1'(Morus abla L.cv.Longsang 1)mulberry leaves in Heilongjiang Province(China)and assess its obesity-preventing/relieving effects.A total of 174 compounds including quercetin,chlorogenic acid,1-deoxyecomycin(1-DNJ)related to antihyperlipidemia effects were identified from the MLIT powder.MLIT treatment reversed the Lee's index,fat coefficient,and serum biochemical parameters in both the obesity relieving and obesity preventing mice fed with high-fat diet.In the obesity relieving experiment,the relative abundance of Desulfovibrio in mouse feces decreased after both 0.5%and 1%MLIT treatments.In obesity preventing experiments,mouse with different amount of MLIT treatments showed increased relative abundance of Akkermansia,Bifidobacterium and Lactobacillus,while Deferribacteres,Desulfobacterota decreased.The beneficial bacteria in the intestinal tract of mice treated with MLIT increased.This study proved that MLIT had antihyperlipidemia potential via modulating intestinal microbiota in mice.

Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus

Two oligosaccharide fractions(MLO 2-1 and 2-2)were purified from enzymatic hydrolysate of mulberry leaf polysaccharide.The results of simulated digestion showed that MLO 2-2 was a digestible oligosaccharide,which could be degraded by human digestive juice;while MLO 2-1 possessed the non-digestible property in the upper gastrointestinal tract and performed the function by regulating the gut microbiota.Hence,MLO 2-1 was selected for the further analysis.The structure of MLO 2-1 was elucidated as follow:α-T-Glcp-(1→3)-β-Glcp-(1→5)-α-Araf-(1→5)-α-Araf-1→5)-α-Araf-(1→3)-α-(2-OAc)-Glcp-1.The in vitro fecal fermentation results showed that MLO 2-1 could modulate the composition of gut microbiota.Meanwhile,MLO 2-1 was effectively metabolized by fecal bacteria to produce lactate and short chain fatty acids,especially acetate and butyrate.The specific metabolic pathways of MLO 2-1 by gut microbiota were further illuminated.Gut microbiota analysis revealed that MLO 2-1 selectively promoted the growth of Ligilactobacillus murinus,a commensal bacterium presented a reduced level in T2DM mice.Animal experiments indicated that MLO 2-1 and L.murinus exhibited hypoglycemic activities.These results demonstrated that MLO 2-1 might alleviate T2DM by selectively accelerating the proliferation of L.murinus.

Response Mechanisms to Flooding Stress in Mulberry Revealed by Multi-Omics Analysis

Abiotic stress,including flooding,seriously affects the normal growth and development of plants.Mulberry(Morus alba),a species known for its flood resistance,is cultivated worldwide for economic purposes.The transcriptomic analysis has identified numerous differentially expressed genes(DEGs)involved in submergence tolerance in mulberry plants.However,a comprehensive analyses of metabolite types and changes under flooding stress in mulberry remain unreported.A non-targeted metabolomic analysis utilizing liquid chromatographytandem mass spectrometry(LC-MS/MS)was conducted to further investigate the effects of flooding stress on mulberry.A total of 1,169 metabolites were identified,with 331 differentially accumulated metabolites(DAMs)exhibiting up-regulation in response to flooding stress and 314 displaying down-regulation.Pathway enrichment analysis identified significant modifications in many metabolic pathways due to flooding stress,including amino acid biosynthesis and metabolism and flavonoid biosynthesis.DAMs and DEGs are significantly enriched in the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways for amino acid,phenylpropanoid and flavonoid synthesis.Furthermore,metabolites such as methyl jasmonate,sucrose,and D-mannose 6-phosphate accumulated in mulberry leaves post-flooding stress.Therefore,genes and metabolites associated with these KEGG pathways are likely to exert a significant influence on mulberry flood tolerance.This study makes a substantial contribution to the comprehension of the underlying mechanisms implicated in the adaptation of mulberry plants to submergence.

Synergistic effects of three traditional herbs green tea,mulberry leaf and corn silk on glucose uptake level of L6 myoblasts and the hypoglycemic mechanism

Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.

Identification of Mulberry Bacterial Blight Caused by Klebsiella oxytoca in Bazhong,Sichuan,China

To provide a scientific basis for controlling mulberry bacterial blight in Bazhong,Sichuan,China(BSC),this study aimed to isolate and purify pathogenic bacteria from diseased branches of mulberry trees in the region and to clarify their taxonomic status using morphological observation,physiological and biochemical detection,molecular-level identification,and the construction of a phylogenetic tree.A total of 218 bacterial strains were isolated from samples of diseased mulberry branches.Of these,7 strains were identified as pathogenic bacteria based on pathogenicity tests conducted in accordance with Koch’s postulates.Preliminary findings from the analysis of the 16S rRNA sequence indicated that the 7 pathogenic bacteria are members of Klebsiella spp.Morphological observation revealed that the pathogenic bacteria were oval-shaped and had capsules but no spores.They could secrete pectinase,cellulase,and protease and were able to utilize D-glucose,D-mannose,D-maltose,and D-Cellobiose.The 7 strains of pathogenic bacteria exhibited the highest homology with Klebsiella oxytoca.This study identifies Klebsiella oxytoca as the causative agent of mulberry bacterial blight in BSC,laying the foundation for the prevention and control of this pathogen and further investigation into its pathogenic mechanism.

Quality Evaluation of Fermented Mulberry Wine

[Objectives]This study to conducted to clarify various nutritional components and the composition and contents of aromatic substances in mulberry wine prepared by fermentation,and promote the quality improvement and product development of mulberry wine.[Methods]Mulberry wine was prepared with mulberry Shengguo 1 as the material using Saccharomyces cerevisiae,and its contents of total sugars,total phenols,total acids,resveratrol,protein,dry extract,ethanol content,Fe,methanol,sulfur dioxide,volatile acids,total flavonoids and proanthocyanidins were determined.Aromatic substances were extracted by n-hexane,and their components and relative contents were determined by gas chromatography-mass spectrometry(GC-MS).[Results]The physical and chemical indexes of the mulberry wine prepared by fermentation were in accordance with the national standards,and it was rich in resveratrol(10.5±0.6 mg/L),protein(381±11 mg/L),total flavonoids(406±18 mg/L)and proanthocyanidins(855±22 mg/L),which were 8.95,19.49,3.76 and 1.63 times higher than those of grape wine,respectively.The aromatic substances were mainly composed of ethyl formate(9.9%),ethyl acetate(7.6%),acetal(6.2%),propanol(5.7%),isobutanol(5.6%)and isoamyl acetate(5.3%).[Conclusions]The results of this study demonstrated that mulberry wine was rich in active substances,which suggested that mulberry wine has great potential in the fruit wine market.

Effects of Nitrogen Amount and Nitrogen Form on 1-deoxynojimycin Content in Mulberry Leaf

[Objective] Effects of nitrogen amount and nitrogen form on 1-deoxynojimycin（DNJ）content in mulberry leaf was explored in order to provide a reference for improving DNJ content in mulberry leaf and its medicinal value.[Method] Effects of nitrogen amount（NO-3-N and NH＋4-N）and nitrogen form（NO-3-N/NH＋4-N）on DNJ content in mulberry leaf were studied by changing nitrogen amount and nitrogen form in water culture solution.[Result] DNJ contents in mulberry leaf both increased at first and then decreased with the increase of nitrogen amount,furthermore compared with NH＋4-N,NO-3-N was more beneficial to the accumulation of DNJ;DNJ content in mulberry leaf obviously increased at first and then decreased with the decrease of NO-3-N/NH＋4-N,among them when its rate was 25/75,DNJ content reached the highest.[Conclusion] Proper nitrogen amount and nitrogen form could effectively improve the DNJ content in mulberry leaf.

Effects of N,P and K Fertilizers on Photosynthetic Characteristics of Mulberry

[Objective] The aim was to research the effects of N, P and K fertilizers on photosynthetic characteristics of mulberry. [Method] In the test, Chuan 826, a va- riety of mulberry, was taken as an example to explore the relationships of chloro- phyll content, leaf area index, net photosynthetic rate, stomatal conductance of leaf, CO2 concentration of cells in leaves and transpiration rate with N, P and K fertilizers at different quantities. [Result] N, P and K fertilizers at different quantities have ef- fects on photosynthetic indices. In detail, N, P and K fertilizer at appropriate quanti- ties enhanced LAI, Chla＋b, Pn, Cond and Tr, Ci of mulberry leaves in control group without fertilizers was significantly higher than that in treatment groups. N, P and K at proper quantities （N at 600 kg/hm2, P at 210 kg/hm2 and K at 300 kg/hm2） would keep functional leaves well and higher photosynthetic period for a long term. [Conclusion] The research explored adjustment of photosynthetic characteristics and differ- ent treatments on photosynthesis of mulberry, providing reference for construction of excellent and highly-qualified mulberry garden.

Induced Expression of Cytochrome P450 CYP305 B1V1 Gene in Different Tissues of Wild Mulberry Silkworm(Bombyx mandarina)

[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA sequence of CYP305 B1 V1 Gene published in GenBank for wild mulberry silkworm, one pair of primers was designed, and the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm treated by NaF, rutin, cypermethrin and ecdysone was also analyzed by the semi - quantitative RT - PCR. Furthermore, homology comparison and phylogenetic analysis for amino acid sequences of this gene were studied. [ Result] Rutin, cypermethrin and NaF had effects on the expression of P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm, while ecdysone had no significant effect. Homology comparison for amino acids indicated that the amino acid sequence of this gene was the most similar to that of CYP305 B1 gene in Bombyx mori with 100% amino acid identity, and highly similar to those of Tribolium casmneum CYP305A1, Apis mellifera CYP305A1, Drosophi- la melanogaster CYP305A1, Anopheles gambiae CYP305A2and Culex pipiens quinquefasciatus CYP2LI. [ Conclusion] CYP305 B1 V1 Gene of wild mulberry silkworm is likely to mainly take part in the metabolism of exogenous compounds, which is of great significance for revealing the function of cytochrome P450 and the metabolic mechanism of different drugs.

Effects of Exogenous Hormones on Subculture Multiplication and Root Induction of Tissuecultured Seedlings of Mulberry

Objective The purpose of this experiment was to investigate the effects of exogenous hormone factors on subculture multiplication and root induction of tis-suecultured seedlings in mulberry. [Method] The aseptic seedlings of Guisangyou 12 were used as the materials and the comparative experiment on subculture multi-plication and root induction used different kinds,concentrations and combinations of exogenous hormones. [Result] The hormone combination of 0.1 mg/L IBA＋1.5 mg/L 6-BA＋0.03 mg/L TDZ showed the best effects to bud multiplication, and the multiple of bud multiplication was 5.28. The effects of NAA was better than IBA and IAA on root induction. The hormone combination of 2.0 mg/L NAA＋1.0 mg/L PP333 was most suitable to the root induction, in which the rooting rate arrived to 100%, the root number was 7.01 and the root length was 1.38 cm on average. [Conclusion] The results wil provide some technical reference for large-scale propagation of mul-berry seedlings in vitro.

Effects of Balanced Fertilization on Mulberry Leaf Quality and Its Active Substance Content

This paper study the effect of nitrogen （N, X1）, phosphorus （P, X2） and potassium （K, X3） in different amounts on crude protein, soluble sugar, total flavonoid and 1-deoxynojirimycin contents in mulberry leaves, with mulberry trees in spring and autumn as the material and as per ＂3414＂ experimental design. The results showed that the qualities and active substance content of mulberry leaves changed from increasing to decreasing with its development; crude protein and solu-ble sugar achieved the peak on August 20; total flavonoid and 1-deoxynojirimycin was the highest on May 15. Fertilizations with N, P and K fertilizers at different amounts had significant effects on quality of mulberry leaf and content of active substances. Specifically, as fertilizer amount increased, the content of active sub- stances grew dramatically and achieved the highest at level 2 （X^2X=X~_）. Based on fertilizer effect functions of objective yield, the recommended amounts of N, P and K fertilizers based on crude protein, soluble sugar, flavonoid content and DNJ in test sites were 718.46, 220.11 and 305.23 kg/hm2, when the highest of crude protein in mulberry leaf was 1 813.83 kg/hm2. When N, P and K fertilizers were recommended at 666.54, 204.41 and 243.18 kg/hm2, soluble sugar in mulberry leaf achieved the peak at 1 042.65 kg/hm2. When N, P and K fertilizers were at 675.96, 326.49 and 462.90 kg/hm2, flavonoid content achieved the maximum at 147.90 kg/hm2. When N, P and K fertilizers were at 720.9, 225.11 and 323.63 kg/hm2, DNJ content was the highest at 13.55 kg/hm2.

Study on Rational Fertilization Scheme for Mulberry(Morus alba Linn) Used for Tea

[Objective] This study was conducted to investigate effects of application amounts of N, P and K and their ratio on tea quality components in mulberry leaves. [Method] Effects of sole application and combined application of N, P and K on the yield and tea quality components of spring mulberry leaves were studied by 3-factor 3-level experiment, based on previous experiments. [Result] Combined ap- plication of 3 of N, P and K was better than sole application and combined appli- cation of 2 from them. Insufficient or excessive fertilization both significantly reduced yield of mulberry leaves and contents of tea quality components, such as 1-de- oxynojimycin （DNJ）, flavonoids, tea polyphenols and amino acids. With the yield of spring mulberry leaves as a target, it is advised to apply N, P and K at rates of 240, 150 and 60 kg/hm^2 , respectively. [Conclusion] This study provides reference and theoretical basis for the establishment of raw material base of high-quality mul- berry tea.

Effect of Nitrogen, Phosphorus and Potassium on the Content and Yield of DNJ of Mulberry Leaves in Spring

[Objective] This study aimed to scientifically improve the 1-deoxynojimycin （DNJ） yield of mulberry leaves in spring. [Method] ＂3414＂ experimental design was adopted to investigate the effects of different fertilizer application rates of nitrogen, phosphorus and potassium on the DNJ content in mulberry leaves. The regression relationship was further simulated with DNJ yield as the objective function and fertil- izer application rates of nitrogen, phosphorus and potassium as the regulatory factors. [Result] The results showed that nitrogen, phosphorus and potassium all have great effects on the DNJ content in mulberry leaves at different parts of mulberry tree （Morus alba）; the DNJ content in mulberry leaves increases with the increasing single-factor fertilizer application rate and reaches the maximum under N2P2K2 level; different fertilizer application rates and combinations of nitrogen, phosphorus and potassium can significantly affect the DNJ content in mulberry leaves; according to the simulated fertilizer effect function, the optimal economic yield of total DNJ in mul- berry leaves was 23.23 kg/hm^2, the recommended fertilization indicators were 286.11 kg/hm^2 N, 95.22 kg/hm^2 P2O5 and 159.62 kg/hm^2 K20. [Conclusion] After confirmation in productive practice, this optimal fertilization program can provide reference for the construction of mulberry plantation with high yield and high quality of DNJ.

Effects of Nitrogen,Phosphorus and Potassium Application on Growth and Dry Matter Accumulation in Mulberry

In this paper, the effect of different fertilizer treatments on the main indexes of growth and development and dry matter accumulation of perennial mulberry, which is cut and pruned in summer, were studied with ‘3414’ field experiment design. The results showed that N, P, and K at proper amounts could promote mulberry growth and development, improve its dry matter content, increase its dry matter accumulation. The effects of N, P, K on leaf yield per plant were respectively expressed as follows： N2 N3 N1 N0 , P2 P1 P3 P0 , K2 K1 K3 K0 . It can be concluded that the recommended fertilizer amounts of dry matter accumulations in mulberry leaf were 694.36 kg/hm2 of N, 198.15 kg/hm2 of P, and 274.26 kg/hm2 of K and the dry matter achieved the maximum at 8 045.04 kg/hm2 . The recommended fertilizer amounts of accumulated dry matter in branch were 1 000.05 kg/hm2 of N, 242.04 kg/hm2 of P, and 218.01 kg/hm 2 of K, and the dry matter achieved the maximum at 5 969.05 kg/hm2 . The recommended fertilizer amount in young shoots were 883.76 kg/hm2 of N, 204.48 kg/hm2 of P and 426.59 kg/hm2 of K and dry matter achieved the maximum at 1 410.24 kg/hm2 . This paper could provide reference for the construction of highly-qualified mulberry field in Sichuan hilly area.

Effect of Mulberry(Morus alba Linn) on Soil and Water Conservation

Rational development and utilization of hilly land is an effective measure for raising land productivity, developing countryside economy, increasing peasants＇ income, conserving soil and water, and improving eco-environment. The growth and root distribution of mulberry trees were studied. Various indexes including soil physical characters and hydrological status, water infiltration rate, water and soil loss and soil erosion amount were tested. The results indicated that mulberry trees had the effects of intercepting rainfall, improving soil infiltration rate, decreasing soil erosion amount, and conserving water and soil

Research of Ecological Restoration of Mycorrhizal Mulberry in Karst Rocky Desertification Area

Mycorrhizal mulberry is a plant, whose mulberry root and Arbuscular Myc-orrhizal fungi came into being a symbiotic relationship by a mean of natural or artifi-cial inoculation. Mycorrhizal mulberry coupling with main ecological barriers, such as barren soil drought in rocky desertification area, wil be beneficial to recovering the karst degraded ecosystem by removing these stresses. So, mycorrhizal mulberry is considered a new effective way in ecological control of karst rocky desertification. The research summarized the structures and the characteristic eco-physiological functions of arbuscular mycorrhizal fungi （AMF）, and preliminarily discussed its availability and research in karst rocky desertification restoration ,including investiga-tion of AMF diversity, the key affecting factor of AMF species diversity, the sym-biosis mechanism between AMF and mulberry, promoting the development of agri-cultural economy in Karst Rocky Desertification area by mycorrhizal mulberry. These elaborate efforts are to demonstrate mycorrhizal mulberry adapting to karst environ-ment from a new view, lay a theoretical basis for karst degraded-ecosystem restoration, and final y guarantee sustainable development of mycorrhizal mulberry in rocky desertification area.

Screening of Field Efficacy of Special Pesticides for Mulberry and Residual Toxicity of Silkworm

[Objective] The paper was to screen the pesticides with good control effects against pests in mulberry and small impact on silkworm. [Method] The combined method of indoor toxicity test, field efficacy test, fast testing of pesticide residues and residual toxicity determination were adopted to carry out test. [ Re- suit] Indoor toxicity results showed that LC50 of mulberry No. 1 against Hyalopterus amygdale was （316.1±57.3） mg/kg, and the control effect of mulberry No. 1 at 14 d reached 62.58% ; fast testing of pesticide residues at 7 d showed that the mulberry leaves were safe to silkworms, which reached the standard of leaves for silkworm. [ Conclusion] Mulberry No. 1 was recommended to be the Special pesticide to apply in mulberry field.

Scheme of Nitrogen Reasonable Fertilization in Mulberry Gardens of Sichuan Hilly Areas

Abstract In the research, five test groups were set, including urea at O, 120, 240, 360 and 480 kg/hm^2, respectively, and analysis was conducted on mulberry leaf yield and some characters of the quality in order to establish utility functions of ap-plied N in mulberry gardens with different production purposes and to provide schemes of rational N fertilization. The results indicated that yield of mulberry leaf and content and yield of active nutritional substances in the leaves would be reduced either by insufficient or excessive N quantity. In addition, 240 kg/hm^2 of applied N significantly improved active nutritional substances in leaf and 360 kg/hm^2 of applied N significantly improved yields of mulberry leaf and active nutritional sub-stances. Therefore, utility functions of applied N quantity with leaf yield, DNJ content and yield were established accordingly. The optimal N quantity in mulberry gardens for sericulture is recommended 360 kg/hm^2 and the optimal N quantities in mulberry gardens for increase of yield and content of DNJ are 299.56 and 294.72 kg/hm^2, respectively.

Observation and Comparison of Pollen Submicroscopic Morphology of Seven Mulberry(Morus alba Linn.) Cultivars

[Objective] This study aimed to investigate the submicroscopic structure and morphological characteristics of mulberry pollen. [Method] Mulberry pollen of seven cultivars were treated by glutaraldehyde fixing, alcohol gradient dehydration, freeze-drying and metal plating to observe the size and surface ornamentation under the scanning electron microscope （SEM）. [Result] The pollen surface characteristics and ornamentation were clearly visible, indicating that this method is suitable for SEM observation of mulberry pollen; experimental mulberry pollen was nearly spher- ical, with two germination apertures and processes on aperture membrane; polar axis was 15.99-22.63 μm long, and equatorial axis was 14.98-20.78 μm long; the pollen volume of ＂Yu No.2＇ was the maximum, while that of ＂Jinxuan No.7＇ was the minimum; the surface of mulberry pollen showed smooth and tumor-like protuberance ornamentations, covered with different densities of particles. [Conclusion] Glutaralde- hyde fixative-alcohol gradient dehydration is an ideal method for morphological char- acteristics of mulberry pollen. This study may provide useful information for palynology identification of Morus plants or even species-level systematic classification.

Establishment of a Mathematical Model for Mulberry Balanced Fertilization in Hilly Areas of Sichuan Province

In order to increase mulberry leaf yield and production efficiency in hilly areas of Sichuan Province through scientific fertilization,＂3414＂,a quadratic regression design for modern fertilization,was employed to simulate the regression relationship between mulberry leaf yield and major fertilizer elements including nitrogen（N）,phosphor（P） and potassium（K） among which mulberry leaf yield was the objective function and N,P and K were the regulatory factors to fertilization level.As a result,7 types of ternary,binary and monadic quadratic fertilizer effect functions of N,P and K fertilizer elements were established.The weight of fertilizer factors in the optimal mulberry leaf yield obtained by various types of functions was in decreasing order of KPKPNPKNKNNP,and the weight of fertilizer factors in the ratio of leaf yield to input was in decreasing order of K〉PK〉NPK〉P〉NK〉NP〉N.To be specific,the ternary quadratic fertilizer effect function（Y=20 313＋0.25X1＋43.48 X2＋34.47X3-0.01X1^2-0.14X2^2-0.06X3^2＋0.05X1X2＋0.02X1X3-0.07X2X3,in which X1,X2 and X3 indicate the fertilizer factors N,P and K,respectively） was a model containing all factors,thereby being a recommended fertilization program for mulberry field.Single factor effect analysis indicated that K fertilizer had the highest influence on mulberry leaf yield and ratio of leaf yield to input.Therefore,among all the simulation functions,the monadic quadratic fertilizer effect function of K fertilizer factor（Y=25 002.27＋44.728X3-0.084X32） leads to the optimal leaf yield and maximum ratio of leaf yield to input.By using the optimal fertilization program with 600 kg/hm2 N,210 kg/hm2 P2O5 and 254.33 kg/hm2 K2O,the optimal economic leaf yield obtained is 30 945.00 kg/hm2,the cost of fertilizer input is 4 403.65 yuan/hm2,and the ratio of leaf yield to input is as high as 17.57.After verification by production practices,the recommended fertilization indicators in the optimal mulberry fertilization program can provide reference for the construction of high-yield mulberry field in hilly areas of Sichuan Province.