In order to explore the effects of mulching measures(such as straw returning and plastic film mulching)on soil aggregates and organic carbon in semi-arid rain-fed agricultural areas.The potato-wheat rotation system in the dryland of Longzhong was used as the research object.The effects of straw stubble mulching(SR),straw strip mulching(SM),plastic film mulching(PM)and open field(CK)on soil organic carbon components(SOC,POC,MAOC),aggregate stability and aggregate carbon contribution rate were systematically analyzed.By evaluating the soil improvement effect of different mulching measures,integrating the intrinsic indicators such as soil organic carbon component dynamics,aggregate stability and crop productivity and water use efficiency data reported in the literature,the optimal mulching mode that can synergistically improve organic carbon and aggregate stability was screened.The results showed that the organic carbon content and aggregate stability of potato and wheat were improved by three mulching measures in potato-wheat rotation,and the effect of SR treatment was the best.Compared with CK treatment,the SOC content of SR and SM treatment increased by 15.38% and 7.78%,and the SOC content of PM treatment did not change significantly.SR and SM treatment increased the POC content in 0—20 cm soil layer by 18.54%,16.62%,and PM decreased the POC content in 0—20 cm soil layer by 3.06%.The overall trend of MAOC increased in each treatment was SR>SM>PM>CK.Compared with CK treatment,SR and SM treatments increased the proportion of MWD,GMD,>2 mm aggregates and SOC content,while PM significantly reduced MWD and GMD.SR increased the SOC enrichment coefficient of>2 mm aggregates by 17.26% in 0—20 cm soil layer,and SR and SM treatments significantly increased the SOC contribution rate of>2 mm aggregates.In summary,under the conditions of this experiment,all three mulching measures can increase soil organic carbon content,and straw stubble mulching returning to the field has the best effect in improving soil organic carbon content and aggregate stability,effectively improving soil fertility.

To clarify the effects of different nitrogen forms combined with magnesium fertilization on potato in the northern foothills of Yinshan Mountains,a field experiment was conducted in Hohhot,Inner Mongolia.The experiment included two nitrogen forms(nitrate nitrogen and ammonium nitrogen)and two magnesium application rates(0,45 kg/ha),resulting in four treatments.The impacts of these treatments on potato yield and its components,dry matter,nitrogen,magnesium,and potassium contents,their accumulation,and nitrogen use efficiency throughout the growth period were analyzed.The results showed that,compared with the application of nitrate nitrogen alone,the combined application of nitrate nitrogen and magnesium significantly increased potato leaf dry matter(21.24%—75.60%)and tuber dry matter(22.04%—62.36%)from tuber initiation stage to harvest stage,as well as whole-plant dry matter(32.54%—70.28%)from seedling stage to harvest stage.It also enhanced nitrogen accumulation(44.01%—69.84%)in tuber initiation stage,starch accumulation stage and harvest stage,whole-plant magnesium accumulation(37.66%—84.21%)during the entire growth period,whole-plant potassium accumulation (24.30%—54.95%) from tuber initiation stage to harvest stage, nitrogen uptake efficiency (30.77%—66.67%) during the entire growth period,and partial factor productivity of nitrogen fertilizer at harvest stage(35.13%).Correlation analysis indicated that potato dry matter,whole-plant nitrogen,magnesium,and potassium accumulation were significantly positively correlated with nitrogen uptake efficiency during the entire growth period(r was all greater than 0.78).At harvest stage,whole-plant potassium content showed a highly significant negative correlation with yield(r was -0.78).Principal component analysis further demonstrated that the total variation among treatments during the entire growth period was highly dispersed,indicating significant differences between nitrogen form combined with magnesium fertilization treatments and those without magnesium application.These findings provide a new strategy for improving nitrogen use efficiency and optimizing nitrogen-magnesium management in potato production in the northern foothills of Yinshan Mountains.

To clarify the basic characteristics and tissue expression pattern of the light signal transduction core bZIP transcription factor StHY5 gene,using potato cultivar Désirée as the experimental material,the full-length CDS sequence of the StHY5 gene was cloned.The sequence characteristics and evolutionary relationship were clarified by bioinformatics analysis.The expression patterns in different potato tissues were analyzed by Quantitative Real-time PCR(qRT-PCR).The results showed that the StHY5 protein consisted of 158 amino acids,with a molecular weight of 17.32 ku and a theoretical isoelectric point of 9.78.It contained one basic leucine zipper(bZIP)domain.The secondary structure of StHY5 was composed of 44.3% α-helices and 55.7% random coils,while the tertiary structure was dominated by α-helices,using the chicken CREB3 bZIP protein as the template.Phylogenetic tree analysis revealed that StHY5 clusters with HY5 proteins from Solanaceae species,showing the closest genetic relationship to CaHY5 in capsicum.qRT-PCR quantification demonstrated that StHY5 was expressed in all tested tissues with significant expression differences,among them the highest expression in stolons,and the lowest expression in tubers.This study clarifies the sequence characteristics and tissue expression pattern of the potato StHY5 gene,providing an experimental foundation for subsequent functional verification and pathway analysis in light signal regulation.

To investigate the effects of salicylic acid(SA)spraying on potato physiology and yield under reduced basal fertiliser conditions,with a view to providing technical reference for hormone regulation to improve potato yield.The potato variety Dongnong 310 was used as the test material,and the field experiment was conducted using the randomised block group method.Five treatments were set:conventional basal fertiliser(T1),50% reduction of basal fertiliser(T2),50% reduction of basal fertiliser+spraying of 6.9 mg/L SA(T3),50% reduction of basal fertiliser+spraying of 13.8 mg/L SA(T4)and 50% reduction of basal fertiliser+spraying of 27.6 mg/L SA(T5).Samples were taken at the early tuber formation,late tuber formation,tuber growth stage and starch accumulation stage,and plant height,stem thickness,relative chlorophyll content,photosynthetic parameters,fluorescence kinetic parameters,canopy coverage and yield were determined.The results showed that under weight loss treatment,spraying salicylic acid could promote potato growth and increase yield per unit area.Among the four SA treatments in the same period,T3 and T4 treatments could increase potato plant height,stem thickness,Net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), photochemical quenching coefficient(qP)and starch content;and T4 and T5 treatments could increase potato canopy coverage,Fv/Fm,maximum electron transport rate(ETRmax), and half-saturated light intensity(Ik).Spraying different concentrations of SA had a significant effect on the yield of potato tuber under the weight-loss treatments,and T1,T3,T4,and T5 treatments were significantly higher than T2,increasing 36.74%,27.35%,29.83%,22.96%.The number of potatoes produced per plant was highest in T1 treatment,followed by T4,and the difference between the two was not significant,and T1,T3,and T4 treatments were 33.47%,28.28%,and 30.29% higher than T2.Single plant yield of T1,T3,T4 and T5 treatments increased by 35.19%,27.87%,28.76%,23.39% compared to T2.The commercial potato rate T3 and T4 increased by 3.67, 3.72 percentage points compared with T2,and the differences were significant.In summary,exogenous SA increased the Pn,Tr and Gs of potato leaves under 50% reduction of basal fertilizer treatment,and increased the yield per unit area,and the spraying of 13.8 mg/L SA in the pre-tuber formation stage could effectively alleviate the adverse effects of reduced basal fertilizer on the growth and yield of potato.

AINTEGUMENTA-LIKE 5(AIL5)is one of the key genes regulating plant growth and development during somatic embryogenesis.To explore the function of the AbAIL5 gene in the somatic embryogenesis of Amorphophallus bulbifer and to accelerate its molecular breeding process,the AbAIL5 gene was cloned from the bulbils of A.bulbifer,and its nucleotide sequence and protein structure were analyzed using bioinformatics tools.The open reading frame(ORF)of AbAIL5 was 654 bp in length,encoding 217 amino acids.Bioinformatics analysis indicated that the AbAIL5 protein was a hydrophilic protein containing a conserved AP2 superfamily domain,without transmembrane domains or signal peptides.Phylogenetic analysis showed that AbAIL5 was most closely related to that of Platanthera zijinensis.Subcellular localization analysis revealed that the protein resides specifically within the cell nucleus.Regarding expression patterns,qRT-PCR analysis revealed that AbAIL5 was expressed in the aerial parts(leaves,petioles,and bulbils)of A.bulbifer,with higher expression levels observed in the early stages of bulbil development.The relative expression levels in petioles,leaves,and bulbils were 1.00,0.55,and 1.42,respectively.For functional verification,an AbAIL5 overexpression vector was constructed and transformed into A.bulbifer callus mediated by Agrobacterium tumefaciens.After 30 days of culture,the AbAIL5 overexpression group showed significantly increased growth rates and germination numbers,as well as an improved overall survival rate compared with the control group.At 70 days,plant growth in the overexpression group was significantly superior to that of the control.qRT-PCR results confirmed that the expression level in the overexpression group was 5.34-fold higher than that of the control.These results indicated that AbAIL5 facilitates adventitious bud differentiation and callus proliferation,providing a theoretical basis for revealing the function of AbAIL5 in the development of A.bulbifer and for improving its genetic transformation efficiency.

In order to reveal the response characteristics of potato growth,development,and photosynthetic traits to different water gradients during the critical growth period in semi-arid regions,this study conducted simulated experiments involving waterlogging (soil moisture maintained at 85% field capacity)and drought stress (drought initiated when soil moisture reached 65% field capacity)to investigate various growth traits and photosynthetic parameters of potatoes.The results showed that the plant height and biomass accumulation of potatoes under waterlogging were not different from those under normal irrigation.The plant height growth and biomass accumulation of potato under drought stress were significantly decreased.Meanwhile,the photosynthetic characteristics of potato leaves were significantly decreased under drought stress,including a 43% reduction in net photosynthetic rate and a 55% reduction in stomatal conductance.There was no significant difference in photosynthetic characteristics(except for stomatal conductance)between waterlogging treatment and normal irrigation treatment.Drought stress and waterlogging treatment decreased the photosynthetic capacity and light energy utilization of potato leaves to different degrees.Among them,under waterlogging conditions,the growth and development of potato leaves were more easily inhibited when exposed to strong light,and the activity of potato leaves was reduced.The growth and development of potato leaves were more easily inhibited when exposed to weak light under drought conditions,but the activity of potato leaves was not reduced.Comprehensive analysis showed that potato was more susceptible to drought stress in tuber expansion stage in semi-arid region,followed by longer waterlogging duration affecting potato growth and development.

To reveal the effects of konjac infection with southern blight disease on the defense enzymes and the phyllosphere microorganism of petiole,the activities of catalase (CAT),peroxidase (POD),superoxide dismutase (SOD),polyphenol oxidase (PPO),and chitinase (CHI) in konjac petioles after inoculation with Sclerotium rolfsii (Sr) were determined using a defense enzyme activity kit (spectrophotometer method),and the structure and diversity of phyllosphere bacterial and fungal communities were investigated using Illumina high-throughput sequencing.The results indicated that the defense enzyme activities in konjac were significantly enhanced after Sr inoculation.Specifically,the activities of CAT and PPO peaked on the third day after Sr inoculation,reaching 6.34 and 5.29 times that of the control group,respectively.The activity of POD continued to increase significantly until the sixth day,reaching 36.98 times that of the control group.The activity of SOD was approximately 1.91 times that of the control group after Sr inoculation,while the activity of CHI was about 5 times that of the control group after Sr inoculation.Analysis of the phyllosphere microbiome revealed that,compared to healthy konjac,the diversity and richness indices of bacterial communities in the leaf petiole phyllosphere showed no significant differences after Sr inoculation,but the richness index exhibited a continuous downward trend.For fungal communities in the leaf petiole phyllosphere,there were no significant differences in diversity and richness indices on the 1st day after Sr inoculation.By the 3rd day,the fungal community diversity index showed no significant difference,while the richness index significantly decreased.By the 6th day,both the diversity and richness indices of the fungal communities were significantly reduced. In healthy A. konjac, the dominant bacterial genera in the petiole phyllosphere were unclassified Enterobacteriaceae, Planomicrobium, and Pseudomonas, while the dominant fungal genera were Phoma, Plectosphaerella, and Cladosporium. On day 1 after Sr inoculation, the dominant bacterial genera shifted to Enterobacteriaceae and Pantoea. By days 3 and 6, the dominant bacterial genus became Alcaligenaceae, and the dominant fungal genus shifted to Sclerotinia. The functional pathways of bacterial communities in the leaf petiole phyllosphere of both Sr-inoculated and healthy konjac primarily included secondary metabolite biosynthesis pathways,microbial metabolism in diverse environments,ABC transport,and two-component system metabolic pathways.The fungal communities in the petiole phyllosphere of healthy konjac mainly consisted of ten functional groups,including endophytes-saprotrophs-lichen parasites-litter saprotrophs-plant pathogens-soil saprotrophs-wood saprotrophs (31.29%),plant pathogens (27.68%),and animal pathogens-endophytes-lichen parasites-plant pathogens-wood saprotrophs (20.27%).As the duration of Sr inoculation increased,the functional groups of fungi gradually became more homogeneous,with the relative abundance of the functional group comprising fungi-leaf saprotrophs-lichen parasites-lichenized plant pathogens-wood saprotrophs continuously increasing.Correlation analysis revealed that the activity of POD,PPO,CAT,and CHI were positively correlated with most phyllosphere bacteria.The activity of POD and CAT activities were negatively correlated with some phyllosphere fungi,while the activity of POD and SOD were positively correlated with the Sclerotium.

To investigate the structure and function of the potato molybdenum cofactor sulfurase(LOS5)gene,StLOS5 was cloned from the potato cultivar Zicai No.3 using PCR,followed by bioinformatics analysis,subcellular localization assays,yeast stress-tolerance assays,and expression pattern analysis under drought stress.The results showed that the CDS region of the potato StLOS5 gene was 2 460 bp in length,encoding 819 amino acids,with a molecular weight of 91.50 ku,a theoretical isoelectric point of 6.78,and a GRAVY value of -0.268,indicating that the protein was hydrophilic.Phylogenetic analysis revealed that the StLOS5 protein had the highest homology with tomato LOS5,suggesting that the two proteins may possess similar functions.Multiple cis-acting elements related to abiotic stress were detected in the promoter region.Subcellular localization showed that StLOS5 was primarily localized in the nucleus.Yeast stress-tolerance assays indicated that StLOS5 enhances tolerance to osmotic stress.Quantitative Real-time PCR analysis demonstrated that StLOS5 exhibits tissue-specific expression,with higher transcript levels in leaves,and responds rapidly to drought stress,with its expression in leaves peaking 12 h after treatment with 8%PEG.These results indicated that StLOS5 plays a role in the potato response to drought stress.

In order to clarify the types of pathogens causing potato common scab in double-cropping cultivation region of Hebei Province,we isolated different types of symptoms tubers which diseased with potato common scab.Morphological identification,PCR molecular identification,and detection of pathogenic genes textAB,necl,and tomA were performed on the isolated strains.The pathogenicity of the strains containing pathogenic genes was further validated using potato chip and radish seedling methods.Finally, the isolated strains were identified at the species level by 16S rDNA sequencing, and a corresponding phylogenetic tree was constructed.The results showed that a total of 6 strains of Streptomyces were isolated from 19 diseased potatoes with different symptom.Through 16S rDNA sequence and amplification analysis,SJZ-1 and CL-1 were identified as S.scabies,SJZ-2 was identified as S.anthocyanicus,CL-2 was identified as S.albogriseolus,SFC was identified as S.thermocarboxydus,QY was identified as S.europaeiscabiei.Strain SJZ-1 and CL-1 contained three pathogenic genes:textAB,necl and tomA.Strain QY only contained the tomA gene.Strain SJZ-2,CL-2 and SFC did not contain any of these three pathogenic genes.The potato chip method showed that QY and SJZ-1 could colonize and produce necrotic spots on small potato chips,the radish seedling method showed that SJZ-1 and QY could inhibit the growth of radish seedlings,while SJZ-2,CL-2,and SFC had no significant pathogenicity in potato chip and radish seedling.This study have preliminary identified the types and composition of potato scab pathogen in double-cropping cultivation region of Hebei Province,and providing a theoretical basis for targeted prevention and potato common scab.

The aim was to explore the role of the small G protein StRab5b in potato resistance to Verticillium wilt.The relative expression levels of the StRab5b gene in the roots of potato tissue culture seedlings(Hutou and Xiapodi)after inoculation with Verticillium dahliae were determined by qRT-PCR.The biomass of V.dahliae in the roots,the lesion area on potato leaves,the content of hydrogen peroxide(H₂O₂),the content of malondialdehyde(MDA),and the activity of antioxidant enzymes were detected in the transgenic potato lines L7,L8,and L10 overexpressing StRab5b(StRab5b-L7,StRab5b-L8,StRab5b-L10)and the non-transgenic plants(CK)after inoculation with V.dahliae.The disease index was calculated using the pot culture method with the StRab5b-L7 transgenic line and the non-transgenic plants(CK).The results showed that the expression of the StRab5b gene was induced in potato after inoculation with V.dahliae.Compared with 0 h after inoculation,the relative expression levels of Hutou and Xiapodi increased significantly by 701.12% and 649.41% at 72 h after inoculation,respectively.After inoculation with V.dahliae,overexpression of StRab5b enhanced the resistance of potato to Verticillium wilt.Compared with CK,the lesion area on potato leaves in StRab5b-L7,StRab5b-L8,and StRab5b-L10 decreased significantly by 78.56%,66.56%,and 59.76% at 96 h after inoculation,respectively;the biomass of V.dahliae in the roots decreased significantly by 73.07%,51.69%,and 23.38% at 168 h after inoculation,respectively;the MDA content decreased significantly by 47.13%,33.08%,and 14.70% at 96 h after inoculation,respectively;the H₂O₂ content increased significantly by 83.67%,48.38%,and 20.15% at 72 h after inoculation,respectively,and the CAT activity increased by 53.54%,33.28%,and 12.62%,respectively;the SOD activity increased significantly by 53.01%,36.36%,and 16.54% at 48 h after inoculation,respectively;the POD activity increased by 55.58%,27.18%,and 1.73% at 96 h after inoculation,respectively;the APX activity increased significantly by 63.69%,44.27%,and 17.11% at 72 h after inoculation,respectively.Compared with CK,the disease rate and disease index of the StRab5b-L7 transgenic line decreased significantly by 43.33 percentage points and 86.07%,respectively.In conclusion,overexpression of the StRab5b gene enhanced the resistance of potato to Verticillium wilt.

To investigate the function of StIMPα in potato, this study used the potato cultivar Kexing No.1 as material and successfully cloned the StIMPα2 gene via PCR. Bioinformatic analysis revealed that the coding sequence (CDS) of StIMPα2 had a length of 1 590 bp, encoding a protein containing the typical domains of the IMPα family. Phylogenetic tree analysis indicated that StIMPα2 was most closely related to AtIMPα-1 and AtIMPα-2 from Arabidopsis thaliana, suggesting functional conservation. Furthermore, analysis of the StIMPα2 promoter region identified multiple cis-acting elements associated with responses to biotic and abiotic stresses. To determine its subcellular localization, a StIMPα2-GFP fusion expression vector was constructed and transiently expressed in leaves of Nicotiana benthamiana via Agrobacterium-mediated transformation. Confocal laser scanning microscopy showed that the GFP fluorescence signal was specifically enriched in the nucleus, confirming that StIMPα2 is a nuclear-localized protein. Expression pattern analysis demonstrated that StIMPα2 expression was significantly induced by abiotic stresses such as low temperature, high salinity, and drought, as well as by BTH (benzothiadiazole). For functional validation, StIMPα2 was overexpressed in N. benthamiana via Agrobacterium infiltration, followed by inoculation with Phytophthora infestans. Pathological phenotype analysis showed that compared with the control, the lesion area on leaves overexpressing StIMPα2 was significantly reduced. Meanwhile, Quantitative Real-time PCR detection of P. infestans biomass confirmed a significant decrease in pathogen biomass in StIMPα2-overexpressing plants. In conclusion, these results indicate that StIMPα2 is a nuclear-localized protein induced by various biotic and abiotic stresses, and it enhances resistance to P. infestans by positively regulating plant immune responses.

To explore the expression pattern of potato gene StEXLB1a and provide a theoretical basis for the resistance to potato disease,based on the previous research of the Laboratory of Plant Resources and Molecular Biology at Northeast Agricultural University,the study using the Atlantic variety of potato as material, cloned the StEXLB1a gene, and preliminarily analyzed its bioinformatics, expression pattern, and disease resistance. The results showed that the full-length cDNA sequence of StEXLB1a gene was 768 bp,encoding 255 amino acids.The annotation indicated that it belonged to the extend protein family genes.Subcellular localization showed that the protein was localized to the cell wall.Among different tissues of potato,StEXLB1a gene expression was highest in leaves,followed by roots and stems,and lowest in flowers and tubers.The expression level of StEXLB1a gene changed significantly under various stress treatments,such as hormone(indoleacetic acid,gibberellin,abscisic acid),stress(high temperature,low temperature,salt,drought),fungal disease dry rot(Fusarium avenaceum),bacterial disease soft rot(Erwinia carotovora subsp,Ecc)and bacterial wilt(Ralstonia solanacearum,RS).The overexpressed transgenic potato plants were inoculated with the dry rot pathogen Fusarium avenaceum,and the transgenic plants were more seriously infected than the wild-type plants.The enzyme activities of active oxygen scavenging system in overexpressed plants were significantly changed,POD and SOD activities were inhibited,MDA content was higher than that of wild-type plants,and the damage degree of plants was aggravated.The results showed that transgenic potato plants with overexpression of StEXLB1a had lower resistance to dry rot than those with wild type.

Catalase(CAT)plays a critical role in plant resistance against biotic stresses.To investigate the functions of StCAT1 and StCAT3 in potato defense against Alternaria solani infection,this study cloned the cDNA sequences of StCAT1 and StCAT3 via RT-PCR.Sequencing results demonstrated that both genes were 1 479 bp in length,encoding 493 amino acids.Phylogenetic tree construction revealed that StCAT1 shared high homology with SlCAT1 from tomato,while StCAT3 exhibited high homology with both SlCAT3(tomato)and SpCAT3 from wild tomato.Following A.solani infection for 6 days,the expression of StCAT1 and StCAT3 was significantly upregulated by approximately 3.9-fold and 8.7-fold,respectively.A co-silencing vector targeting both StCAT1 and StCAT3 was constructed,and transient co-silencing was achieved using virus-induced gene silencing(VIGS).qRT-PCR screening identified an effective co-silenced potato line(VIGS-3)with suppressed StCAT1 and StCAT3 expression.Upon A.solani inoculation,the silenced line exhibited 42% larger lesion areas compared to the control,indicating that co-silencing StCAT1/StCAT3 compromised potato resistance to early blight.DAB(3,3'-diaminobenzidine)staining further demonstrated that H₂O₂ accumulation in co-silenced leaves was significantly elevated,with staining intensity being 3.8-fold higher than that in the control.These results demonstrate that StCAT1 and StCAT3 regulate H₂O₂ homeostasis to mediate potato resistance against A.solani,providing a theoretical foundation for elucidating the molecular mechanisms of CAT genes in potato-pathogen interactions.

To investigate the molecular mechanisms of the potato miR7997 family in response to Phytophthora infestans infection,this study analyzed the sequence characteristics,target gene prediction,expression patterns,and stress-responsive expression dynamics of Stu-miR7997 and its targets using the potato cultivar Desiree.The results revealed that the potato miR7997 family comprised three members(Stu-miR7997a/b/c)distributed across two chromosomes,with Stu-miR7997a/b sharing identical mature sequences.All precursor sequences formed canonical stem-loop secondary structures,with minimum folding free energies ranging from -37.00 to -49.50 kcal/mol,and mature sequences were located on the 5' arm.Stu-miR7997c exhibited distinct sequence length and functional element distribution compared to Stu-miR7997a/b.Promoter analysis identified light-responsive,hormone-responsive,transcription factor-binding,and stress defense-related cis-regulatory elements;target prediction identified 19 genes predominantly co-regulated by the miR7997 family.Tissue-specific expression profiling showed that Stu-miR7997a/b were highly expressed in stems,while Stu-miR7997c accumulated predominantly in roots.Upon P.infestans infection,all miR7997 members were significantly downregulated,whereas their target genes-including the transcription factor MYB92 gene,NAD(P)H-quinone oxidoreductase gene,and pectin lyase gene-were markedly upregulated.These findings suggest that the Stu-miR7997 family may indirectly modulate potato disease resistance by negatively regulating target genes.This study provides a theoretical foundation for further exploration of the miR7997 family in potato late blight resistance.

The TCP transcription factor family is unique to plants and plays a crucial role in key biological processes such as plant growth,metabolic activities,and stress responses.To investigate the quantity,distribution and expression patterns of TCP gene family in the Amorphophallus konjac genome,bioinformatics approaches were utilized to identify the AkTCP gene family,followed by a comprehensive analysis of the physicochemical properties,subcellular localization,collinearity,gene structure,and evolutionary relationships.Additionally,gene expression in response to biotic and abiotic stresses was examined by qRT-PCR.The findings were summarized as follows:the A.konjac genome contained 30 AkTCP family genes,with protein lengths ranging from 135 to 562 amino acids.These proteins had molecular weights between 15.08 and 57.20 ku,and isoelectric points between 4.96 and 11.49.Most of these were basic,hydrophilic,and unstable proteins,predominantly localized in the nucleus.These genes were unevenly distributed across the chromosomes.The collinear AkTCP genes were unevenly distributed across eight chromosomes,comprising four inter-chromosomal and five intrachromosomal collinearity events.The AkTCP gene structure was relatively simple,with most genes lacking introns,and all AkTCP transcription factors exhibit a highly conserved TCP domain.Phylogenetic classification separates the 30 AkTCP genes into two major classes,Class Ⅰ and Class Ⅱ,with Class Ⅱ further subdivided into the CIN and CYC/TB1 subclasses.The promoters of AkTCP genes contain cis-elements related to physiological response,light-responsive,phytohormone-responsive and stress-responsive elements.The results of qRT-PCR analysis indicated that AkTCP family members were involved in and responded positively to drought stress, salt stress, methyl jasmonate and soft rot pathogens of A. konjac.

To understand the changes in endogenous hormones and related gene expression levels after konjac infection with southern blight disease and reveal the main hormone pathways involved in konjac's response to southern blight disease,the ultra performance liquid chromatography (UPLC) and tandem mass spectrometry (MS/MS) was employed to detect the changes in the contents of phytohormone (auxin,abscisic acid,trans-zeatin nucleoside,jasmonic acid,and salicylic acid) in the three-month-old Amorphophallus muelleri,which was infected with southern blight disease for 0,1,3 and 6 days.Moreover,the gene expression levels of abscisic acid,jasmonic acid,and salicylic acid pathways was analyzed by Real-time Quantitative PCR.The results showed that southern blight disease caused changes in the levels of various endogenous hormones in konjac.The content of indole-3-acetic acid tended to increased and then decreased with the southern blight disease infection,while the content of indole-3-butyric acid showed tended to decrease;the content of trans-zeatin nucleoside significant decreased with the infection of southern blight disease;the content of abscisic acid showed a significant increase followed by a decrease with the infection of southern blight disease;the content of jasmonic acid metabolites and salicylic acid metabolites were significantly increased than the control group with the infection of southern blight disease.The content of jasmonic acid of konjac infected with southern blight disease 1,3,and 6 d was 2.31,2.31,and 5.08 times that of the control group,and the content of salicylic acid was 5.53,4.60,and 7.38 times that of the control group,respectively.Eight genes related to abscisic acid,jasmonic acid,and salicylic acid pathways were activated and participated in the response process of konjac to southern blight disease.The above results indicated that the endogenous hormone homeostasis was disrupted after konjac infection with southern blight disease,activated the expression of plant hormone pathway related genes,and jasmonic acid and salicylic acid might play an important role in konjac's resistance to southern blight disease.

PHR1 is a crucial factor in balancing plant disease resistance and low phosphorus stress resistance.To investigate the nature and function of the StPHR1 gene in potato and to explore the role of StPHR1 in the process of potato resistance to Alternaria solani infection,the CDS sequence of the StPHR1 gene was cloned by PCR technology using potatoes as the research material,and the structural,physicochemical properties,and phylogenetic relationships of StPHR1 were analyzed and predicted using bioinformatics software,then,the expression level of StPHR1 during the infection of potatoes by A.solani and under different hormone treatments was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the CDS of the StPHR1 gene was 1 353 bp,encoding 450 amino acids.The protein had a molecular formula of C₂₁₄₇H₃₃₉₉N₅₉₅O₇₁₁S₁₈,a molecular weight of 49.51 ku,and a theoretical isoelectric point of 5.07,encoding a hydrophilic,unstable protein with no signal peptide and no transmembrane structure.Its secondary structure consisted mainly of random coil and α-helix.Phylogenetic tree analysis revealed that the StPHR1 protein was most closely related to Arabidopsis thaliana; conservative domain analysis revealed that the StPHR1 protein,like other PHR1s,possesses both MYB-CC and MYB conserved structural domains at its C-terminus.Relative expression analysis found that StPHR1 was significantly induced by A.solani and salicylic acid,and it was hypothesized that StPHR1 played an important role in A.solani infection of potato and in the response to salicylic acid; and the subcellular localization showed that the StPHR1 protein was localized in the nucleus.It is hypothesized that StPHR1 may regulate potato resistance to A.solani through its MYB transcription factor activity and response to salicylic acid.

Chalcone synthase(CHS)is an important structural gene that regulates the early biosynthesis of flavonoid pathways in plants,and plays a role in plant growth and development and stress response.Previously,the key genes StCHS4 and StCHS5 for anthocyanin biosynthesis were identified in the potato CHS family by expression analysis.To further explore the function of potato StCHS4 and StCHS5 in the biosynthesis of flavonoids and anthocyanins,the characterization of StCHS4 and StCHS5 proteins was analyzed by online website.The 35S∷StCHS4-GFP and 35S∷StCHS5-GFP recombinant vectors were constructed by homologous recombination method based on the pRI101 binary vector,and then were transformed into Agrobacterium GV3101 strain.The subcellular localization of StCHS4 and StCHS5 proteins was determined by transient transformation of Nicotiana benthamiana.N.tabacum was used as the experimental material for transient overexpression and stable genetic transformation to analyze the content of total flavonoids and anthocyanins after overexpression of StCHS4 and StCHS5 genes.The results showed that the secondary structures of StCHS4 and StCHS5 proteins were mainly α-helix and random coil.StCHS4 was an unstable hydrophilic protein,and StCHS5 was a stable hydrophilic protein.The sequence alignment revealed that StCHS4 and StCHS5 had a close relationship with the CHSs of Capsicum annuum and Solanum lycopersicum,respectively.The results of subcellular localization revealed that StCHS4 and StCHS5 proteins were localized in the cytoplasm and cell membrane.In transient overexpression of tobacco,StCHS4 and StCHS5 genes significantly enhanced anthocyanin accumulation at 3—5 days after injection.Three transgenic N.tabacum lines of StCHS4 and StCHS5 gene were obtained,respectively.Compared with the wild type,the expression of StCHS4 and StCHS5 in transgenic plants was significantly higher,and the contents of total flavonoids and total anthocyanins were higher than those in the wild type.The total flavonoid content in StCHS4-OE3 and StCHS5-OE1 transgenic plants was significantly increased.The anthocyanin content in StCHS5-OE1 and StCHS5-OE2 plants increased by 89%,131%,respectively.The above results demonstrated that StCHS4 and StCHS5 were the key CHS genes in the flavonoid pathway of Solanum tuberosum,and the overexpression of StCHS4 and StCHS5 contributed to the biosynthesis of anthocyanins and flavonoids.

In order to investigate the effects of reactive oxygen species(ROS)on the growth and antioxidant system of sweetpotato under potassium deficiency stress,and further reveal the defense mechanism of ROS regulation in sweetpotato under potassium deficiency stress.This study selected two sweetpotato varieties,Xushu32(low potassium tolerant)and Ningzi 1(low potassium sensitive),as experimental materials.The experiment was conducted in an artificial climate chamber using hydroponic cultivation to ensure consistent environmental conditions.After pretreatment with exogenous H₂O₂ and diphenyl chloroiodate(DPI),ROS inhibitor,the sweetpotato seedlings were subjected to potassium deficiency treatment.The changes in relative H₂O₂ content,antioxidant gene expression levels,and enzyme activity indicators in sweetpotato roots at different time points after potassium deficiency treatment were measured.The morphological characteristics of sweetpotato were recorded to explore the effects of ROS signaling on the growth of sweetpotato seedlings under potassium deficiency stress.The results showed that under potassium deficiency treatment,the biomass of sweetpotato decreased and leaf color lightened.The relative concentration of ROS in sweetpotato seedling roots showed the highest fluorescence intensity on the 14th day.The expression levels of antioxidant-related genes in sweetpotato roots generally increased,with a more significant increase in expression levels after H₂O₂ pretreatment.Compared with-K treatment,both H₂O₂ and DPI pretreatment could enhance the activity of antioxidant enzymes in sweetpotato leaves,and different enzyme activities showed different trends within 21 days of potassium deficiency treatment.There were varietal differences in the effects of different treatments.The experimental results showed that H₂O₂ could act as a stress signaling molecule to increase the expression level of antioxidant genes and the enzymatic activities of APX,SOD and POD in sweetpotato seedlings,thus scavenging excess ROS and effectively alleviating the inhibition of growth of sweetpotato seedlings by potassium deficiency stress.And DPI could inhibit the production of endogenous H₂O₂,maintain the antioxidant enzyme activity,protect the ROS enzymatic scavenging system of sweetpotato seedlings and avoid oxidative damage to tissues.

In order to study the role of phototrapping pigment binding protein gene in potato photosynthesis,the phototrapping pigment binding protein gene StCP24 (LOC102586836)was bioinformatics analyzed and cloned into potato variety Dongnong 310 to obtain transgenic potato.The morphological indexes,leaf physiological indexes,fluorescence parameters,and gene relative expression of transgenic and control lines were measured under different light intensities after clonal propagation.The function of StCP24 gene was analyzed.The effect of this gene on chloroplasts was determined by ultrastructural observation.The results of bioinformatics analysis showed that StCP24 protein was a hydrophilic protein,and the promoter of StCP24 gene contained elements such as photoresponse,defense and stress response.Phylogenetic analysis showed that the amino acids encoded by StCP24 gene in potato were closely related to those encoded by StCP24 gene in cultivated tomato.Under different light intensity treatments,StCP24 gene overexpressed potato significantly increased stem diameter,leaf area,leaf weight,root length,chlorophyll a,b content,Fv/Fm,ETR,ETRmax and qP compared with wild-type potato.The overexpression of StCP24 gene can make the lamellar stacking of thylakoid granules more tightly and to a higher degree.In conclusion,the overexpression of StCP24 gene can increase the content of chlorophyll a and b in leaves,improve the electron transfer rate in photo cooperative photoreaction,and promote the growth of transgenic potato plants.

To explore the expression patterns of potato genes and provide theoretical basis for the subsequent study of gene resistance to potato diseases.Three endogenous small G protein genes StRac5,StRac7 and StRac13 were cloned from potato,and their bioinformatics and expression patterns were analyzed.The results showed that StRac5,StRac7 and StRac13 contained similar conserved sequences to the small G proteins in Arabidopsis and rice,all belonging to ROP proteins.The protein domains of StRac5 and StRac13 were identical with AtRop1,AtRop3,AtRop5,AtRop6 of Arabidopsis and rice OsRac5,OsRac6,OsRac7,and StRac7 was identical with AtRop9.Phylogenetic tree analysis showed that StRac7 belonged to class Ⅱ and was closest to Arabidopsis AtRop9.StRac13 belonged to class Ⅲ and was the closest genetic relative to Arabidopsis AtRop7.StRac5 belonged to class IV and was the closest genetic relative to rice OsRac5 and OsRac6.The relative expression levels of StRac5,StRac7 and StRac13 genes in different tissues of potato were leaf>root>stem.Compared with the expression in stems,the expression of StRac5,StRac7 and StRac13 in leaves were increased by 1 222.4%,2 531.3% and 468.2%,respectively.After inoculation,the relative expression of StRac5,StRac7 and StRac13 genes increased first and then decreased,and the relative expression of StRac5 and StRac13 genes increased by 62.2%,40.4%,respectively,compared with 0 h after inoculation 24 h.The relative expression of StRac7 gene was increased by 827.8% at 72 h after inoculation compared with 0 h.After ABA,SA,GA and 6-BA treatment,the relative expression of StRac5,StRac7 and StRac13 genes showed a downward trend.After JA and IAA treatment,the relative expression of StRac5,StRac7 and StRac13 genes showed an up-regulated trend.Therefore,StRac5,StRac7 and StRac13 genes can respond to potato late blight infection,and their expression patterns are different in different tissues and under different hormone treatments.

In order to investigate how to alleviate the effects of insufficient light in potato production,the potato variety Dongnong 310 was used as the test material,and three shading treatments were set up:normal light(Z₀,blackout rate is 0),single-layer blackout screen covering treatment(Z₁),double-layer blackout screen covering treatment(Z₂).Different concentrations of epbrassinolide(0 mg/L,denoted as CK)were sprayed in the potato tuber formation stage(0.02 mg/L,denoted as E₁;0.10 mg/L,labeled E₂).Three measuring periods were set,the plant height,chlorophyll a,b content,photosynthetic parameters,fluorescence kinetic parameters,and yield per plant were measured and analyzed.The relative expression levels of LHcb genes in 12 leaves were measured.The results showed that Z₁ conditions were more suitable for the growth of Dongnong 310,while Z₀ conditions produced photoinhibition,and the indexes were lower than Z₁.Under the condition of Z₂,the illumination was insufficient,and the indexes were lower than that of Z₁.EBR spray could effectively increase the yield per plant,and E₂ treatment under Z₀ condition significantly increased by 32.62% compared with CK,E₁ and E₂ treatment under Z₁ condition significantly increased by 48.38% and 24.24% compared with CK,respectively,E₂ treatment under Z₂ condition significantly increased by 18.45% compared with CK.In terms of weight of dry matter per tuber,E₂ treatment under Z₀ condition increased by 27.24% compared with CK,and E₁ and E₂ treatment under Z₁ condition increased by 40.09% and 26.56% compared with CK,and the above differences were significant.The relative expression of 12 LHcb genes could be increased by increasing the shading degree and spraying EBR,which was conducive to improving the light energy capture and utilization in leaves.

To clarify the effect of callus treatment on the quality of sweet potato,provide a theoretical basis for high quality storage of sweet potato.Two edible varieties of sweet potato were stored and taken at different storage periods(0,7,15,30,60 and 90 d).Different degree of injury were carried out on them.The experiment was divided into four treatments: CK, high temperature callus,callus after shallow injury,and callus after deep injury.Then put materials in an incubator with relative humidity of 85% at 35 ℃ for 48 h to healing the wound.The diameter of soft rot spots,the activity of epidermal defense enzymes(PPO,POD,PAL)and the content of nutrients in potato pieces were measured.The results showed that the spot diameter of sweet potato root increased with the extension of storage period after inoculation of Rhizopus stolonifera. Sweet potato treated with callus was more resistant to soft rot than sweet potato stored in ordinary environment.After storage for 90 days,the diffusion diameter of soft rot spots of the two varieties after callus treatment with shallow injury was only 74.6% and 71.1% of that of the control treatment.At the early stage of storage, the activity of defense enzymes in each treatment remained at a high level from 0 to 30 d. After 90 d of storage, the activity of defense enzymes after callus injury treatment was significantly higher than that of control treatment. At 60 d of storage, the sucrose content of root tuber of two varieties with treatments of callus after shallow and deep injury was significantly higher than that of CK. Callus treatment of sweet potato after different degrees of injury during storage period can significantly increase the activity of defense enzymes in sweet potato epidermis,reduce the spread speed of soft rot infection,and improve the resistance of sweet potato to soft rot.The degree of injury had influence on the callus effect of sweet potato.The callus after shallow injury was better than that of deep injury in terms of defense enzyme activity and storage quality.

To express the protein StSPS1 encoding potato sucrose phosphate synthase(SPS)in E.coli and prepare polyclonal antibodies.The StSPS1 gene was cloned from the tubers of tetraploid potato variety Chuanyu 10,with a total coding region of 3 165 bp and a protein length of 1 055 aa.Subsequently,based on the constructed His tag fusion expression vector PET30a-StSPS1,the protein induction,denaturation,purification,renaturation,and rabbit immune tests of StSPS1 protein were carried out.The results showed that the molecular weight of StSPS1 protein was approximately 119.62 ku,and its expression was minimal in soluble supernatant,mainly in insoluble precipitates.The optimal induction conditions were induced with 0.5 or 1.0 mmol/L IPTG at 37 ℃ for 4 h.Due to StSPS1 being an inclusion body protein,it was subjected to inclusion body denaturation and purified using His tags to match the size of the target band.At the same time,a protein immunoblotting(WB)test was performed using His antibodies,and the target band was detected at 119.62 ku,indicating the successful purification of StSPS1 inclusion body protein.Finally,by injecting the dialyzed and refolded StSPS1 protein into the subcutaneous tissue of rabbits,two antibodies against StSPS1 were successfully immunized.After WB identification,it was found that both antibodies could hybridize target bands in the antigen and the total protein of leaves in Chuanyu 10.In summary,potato StSPS1 protein was induced and purified,and rabbit polyclonal antibodies against StSPS1 protein were successfully prepared.

Anthocyanins are a kind of important flavonoids produced during the secondary metabolism of plants.Anthocyanidin synthase(ANS)catalyzes the conversion of leucoanthocyanidins to anthocyanidins in the anthocyanin synthesis pathway.Previous studies have shown that the anthocyanidin synthase gene plays an important role during the coloring process of plant organs.To investigate whether the gene can promote anthocyanin synthesis in potato tubers,using the red-skinned and red-fleshed colored potato variety Hongmei as the material,anthocyanidin synthasegene was isolated from tubers of color potato by RT-PCR, the characterization analyzed by bioinformatics and function validated by overexpression into Arabidopsis thaliana.The results showed that sequence 1 406 bp cDNA of the gene was amplified from the potato,including 1 368 bp open reading frame(ORF).The ORF encoded a deduced protein of 455 amino acids.The gene was named as StANS1a by bioinformatics analysis.GenBank accession number is ON512347.The amino acid sequence analysis revealed that the deduced protein contained a DIOX_N'domain located at 52—166 resides and a 2OG_FeⅡ_Oxy domain located at 215—312 resides.Arabidopsis thaliana plants was transformed with constructs overexpression vector StANS1a.The transcriptional expression of StANS1a in transgenic lines were tested by RT-PCR.The results showed that in the same population,the transcriptional expression of StANS1a showed an increase in different transgenic lines compared with the wild-type(Wt).The anthocyanin content was further analyzed in transgenic lines.Compared to the Wt,the anthocyanin contents of the seven lines(T₁—T₇)were increased from 2.17% to 54.61%. These results strongly demonstrated that overexpression of the StANS1a gene could promote anthocyanin synthesis in transgenic plants.The colored potato StANS1a gene plays an important role in the anthocyanin metabolic pathway.

In order to study the effect of different irrigation methods and ridge planting experiments on water use efficiency and yield of potato in dryland,and to identify an optimal ridge planting and irrigation combination mode of potato with high yield and water use efficiency simultaneously,and further provide a theoretical basis for the production efficiency of dryland potato.The two-factor experiment was persistently performed in Lanxian County Shanxi Province during two potato growing seasons(2019—2020).The three ridge planting,including conventional ridge cultivation(M₁),concave ridge cultivation technique(M₂)and outdoor planting(M₃);the two irrigating including hydraulic driving zonal sprinkler(G₁)and drip irrigation(G₂),deeply exploring the effects of ridge planting and irrigation and their interaction effects on water use efficiency and yield of dry potato.The results showed that irrigation and ridge planting had obvious regulation effects on water use efficiency and yield of dry potato.Compared with M₁ and M₃,M₂ could significantly increase soil use efficiency water storage consumption.Under G₁ treatment,the water use efficiency and yield of M₁ and M₂ treatments increased by 14.08%,13.58%,and 23.28%,21.92%,respectively,compared to M₃;under G₂ treatment,the water use efficiency and yield of M₁ and M₂ treatments increased by 11.88%,11.50%,and 22.05%,20.45%,respectively,compared to M₃,and the combination of concave ridge cropping + hydraulic driving zonal sprinkler irrigation treatment combination G₁M₂ had the highest yield.Under the same ridge planting,the water use efficiency and yield of hydraulically driven zonal sprinkler irrigation(G₁)were the highest.Based on the regulating effects of ridge planting and irrigation on water use efficiency and grain yield,the G₁M₂ treatment could help synchronously to improve the grain yield and water use efficiency.This can be an optimal model of ridge planting and irrigation combination treatment of dry potato area of Shanxi Province.

The study aims to understand the biological process and metabolic pathway of konjac under different root separation modes from black locust,in order to provide molecular basis for the expression regulation and response mechanism of related genes in intercropping konjac roots.Illumina NovaSeq 6000 sequencing technology was used for transcriptome analysis of konjac roots separated by plastic membrane(FS),nylon mesh(MS),and non-separation(NS).A total of 59.82 Gb data were obtained,and 92 358 transcript sequences corresponding to 40 122 Unigenes were generated after de novo assembly and quantitative assessment.The most annotated Unigene in the NCBI-Nr database was 23 960,accounting for 59.72% of the total Unigenes.The transcriptome data were pair-wise compared,and the total quantities of differential expression genes of FS_vs_MS,FS_vs_NS and MS_vs_NS were 1 776,733 and 896,respectively.Of which,their corresponding up-regulated genes accounted for 33.1%,22.8% and 50.8%.GO enrichment analysis indicated that significantly differential expressed genes were mainly involved in plasma membrane,cell membrane,cell periphery,cell wall,encapsulating structure,intrinsic components of membrane,and intrinsic components of plasma membrane.KEGG enrichment analysis showed that the ribosome,oxidative phosphorylation and plant hormone signal transduction were the most significant metabolic pathways in the roots of konjac separated by nylon mesh and non-separation compared with plastic membrane.Transcriptome analysis of konjac roots provided the basic data for studying response mechanism of intercropping konjac.

RPP13 is a typical R gene that can induce plant immune response by recognizing of the effectors.The transcriptome sequencing analysis of five potato cultivars infected with Potato spindle tuber viroid (PSTVd)showed that RPP13 genes were up-regulated in the five potato cultivars infected with PSTVd.In order to study whether this gene was related to PSTVd resistance in potato,DNAMAN 8.0 software was used to perform sequence alignment analysis on RPP13 upregulated gene sequences obtained by transcriptome sequencing.The common conserved region was selected as the silencing object,and a stem-loop structure inverted repeat with the conserved region as the arm and the intron from somatic embryogenesis receptor-like kinase 1 (SERK1)(Accession number EF175216)as the loop was inserted in plant expression vector pROKⅡ to construct RNAi vector named pROKⅡ-RPP13.The pROKⅡ-RPP13 vector was transferred into Agrobacterium tumefaciens LBA4404 by freeze-thaw method,and the potato cultivar Minfenghong was transformed by Agrobacterium tumefaciens mediated genetic transformation method. Healthy potato leaves of 28 days of seedling age were used as materials. After 2 days of pre-culture, 10 minutes of infection, 2 days of co-culture under dark conditions, roots and buds were induced by the resistance screening medium. The regenerated plants were screened by PCR, and three positive transgenic plants were obtained.Using efla gene of potato as the reference gene,Real-time RT-PCR was used to detect the expression of RPP13 gene in potato transformed plants.The results showed that the expression level of RPP13 gene in the three transgenic plants obtained was significantly different from that in the control without transgenic.In the obtained RNAi transgenic potato plants,the expression level of RPP13 gene was decreased by 35%—60%.

In order to explore the molecular mechanism of diploid potato material response to drought stress under different drought stress time treatment,and explore drought resistance related genes.The high drought-resistant diploid resource A90 was used as experimental material,and transcriptome sequencing technology was used to analyze the differentially expressed genes under PEG-6000 stress at different times.Differentially expressed genes involved in drought stress response of potato were predicted by GO enrichment,KEGG pathway analysis and transcription factors,and key genes of drought resistance regulation were preliminarily explored.The expression of three candidate genes was analyzed by RT-qPCR.The results showed that there were 2 519 differentially expressed genes in the A90 treated with 20% PEG-6000 for 3,6,24 h compared with the control,and these genes were mainly enriched in plant hormone signal transduction,glutathione metabolism,phenylpropanoid biosynthesis,pentose and glucuronate interconversions and other drought-related processes.In addition,among 340 codifferentially expressed genes significantly enriched in the pathway,53 genes were annotated to 15 transcription factor families including RLKs,AP2/ERF and Tify.Among which StST/K1,StERF1 and StTify1 had higher expression levels,which might be key genes in drought resistance regulation.StST/K1 and StERF1 were mainly up-regulated in roots and leaves under drought and low temperature stress,StERF1 was up-regulated in roots,stems and leaves under salt stress.StTify1 was up-regulated in stems and leaves of the material under drought,and it was up-regulated in roots,stems and leaves under salt stress.But it was down-regulated only in stems of the material under low temperature stress.These results indicate that the three selected genes can respond to drought,salt and low temperature stress,it can provide a theoretical basis for study on drought resistance candidate genes in potato molecular breeding in the future.

In order to explore the important role of SBP1(S-RNase-binding protein 1)gene in plant self-incompatibility,a diploid wild potato was used as the material to obtain the full-length cDNA sequence of potato SpSBP1 (GenBank: MZ803088)by using RT-PCR cloning technique,and performed bioinformatics analysis and CRISPR/Cas9 vector construction for SpSBP1 gene.The results showed that the full length cDNA of SpSBP1 gene was 1 176 bp,containing 92 bp 5' non-coding region and 163 bp 3' non-coding region.The maximum open reading frame(ORF)of SpSBP1 gene was 921 bp,which encoding 306 amino acids.Protein domain analysis showed that SpSPB1 protein included Smc superfamily domain,RING finger domain and Zinc finger domain.The homologous sequence alignment and phylogenetic tree analysis showed that SpSBP1 had the highest homology with S.chacoense,followed by Nicotiana alata.Bioinformatics analysis showed that the molecular weight of SpSBP1 was 34.731 44 ku,with a theoretical isoelectric point of 5.10,and it was speculated that SpSBP1 was an acid unstable hydrophilic protein. The secondary structure predicted that the protein was mainly composed of α helix and random curling. And the protein was a non-secretory protein and had no transmembrane structure. SpSBP1 gene was cloned from wild diploid potato,and CRISPR/Cas9 vector was successfully constructed,and also the genetic transformation was carried out.

In order to clarify the genetic differences of the six new-colored potato strains NNCS-1,NNCS-5,NNCS-6,NNCS-7,NNCS-33 and NNCS-37 at the cytogenetic and DNA molecular level,this experiment used material MIN-021 as control,and the regular staining and slides preparing and SSR marker technology were used to analyze the pollen fertility,chromosome configuration at meiosis metaphase Ⅰ of pollen mother cells and SSR polymorphism of the above six varieties.The results shaved that the pollen fertility of the six new-colored potato strains were 44.00%,66.78%,74.44%,60.70%,78.10% and 80.22%,respectively.And the chromosome pairing configurations of these test materials were 8.45Ⅰ+6.09Ⅱ+7.35Ⅲ+1.33Ⅳ,6.91Ⅰ+7.13Ⅱ+5.73Ⅲ+2.41Ⅳ,5.19Ⅰ+9.16Ⅱ+4.51Ⅲ+2.74Ⅳ,6.55Ⅰ+8.35Ⅱ+4.97Ⅲ+2.46Ⅳ,4.02Ⅰ+10.15Ⅱ+3.40Ⅲ+3.37Ⅳ and 3.31Ⅰ+10.42Ⅱ+2.99Ⅲ+3.72Ⅳ,respectively,indicating that there were certain differences among the strains at the cytological level.Ten suitable SSR primers were selected to amplify the genomic DNA of each new strain and control material,and 151 SSR polymorphic sites were obtained,accounting for 86.78% of the total number of polymorphic loci.The genetic differences among the materials were revealed.In addition,a SSR fingerprint was amplified with the selected specific primers C42,which could identify six new-colored potato strains and control.Based on the average genetic distance(GD=0.62),the seven materials were divided into three categories: the new strain NNCS-37,NNCS-6,NNCS-7,NNCS-5 and the control(MIN-021)belong to one categorie.The new strains NNCS-33 and NNCS-1 were separately classified as a categorie.

To investigate the effect of different field water capacity on potato root growth and improve potato yield in the Northwest. The potato variety Hermes was grown in the field under a rain shelter and film mulching with drip irrigation under six field water capacity gradient treatments(85%-95%,75%-85%,65%-75%,55%-65%,45%-55% and non-irrigation)in 2018 to investigate the effects of different field water capacity on root system characteristics,tuber yield and relevance. The results showed that the root distribution gradually deepened as the field water holding capacity decreased,and when the field water holding capacity was greater than 45%,the roots were mainly distributed at 0-10 cm of the monopoly surface;when the field water holding capacity was less than 45%,the roots were mainly distributed at 20-40 cm of the monopoly side. Tuber yield and large potato rate were higher under the field water capacity of 65%-95% than that under other treatment,and the compensation effect was significant(P<0.05). Among them,the field water capacity of 75%-85% treatment had the highest yield,with an average yield of 884.06 g/plant,which was 257.61% higher than the T6 treatment. Correlation analysis revealed that field water capacity was significantly(P<0.05)positively correlated with top root length,root surface area,large potato rate and tuber yield,and top root length and surface area were significantly(P<0.05)positively correlated with potato tuber yield. In summary,field water capacity greater than 65% significantly increased root length,root surface area and yield on the monopoly surface,with the highest yields achieved under the 75%-85% water capacity treatment.

In order to study the molecular mechanism of sodium silicate enhancing risistance of potato to Rhizoctonia solani,a gene StWRKY11 with high expression level in potato transcriptome induced by sodium silicate was cloned and its bioinformatics was analysed.Total RNA was extracted from potato,amplified by RT-PCR and cloned.Through bioinformatics related software,the structure prediction and prediction analysis were carried out.The results showed that StWRKY11 gene with a open reading frame of 1 005 bp was cloned from potato Atlantic,encoding 334 amino acids.The molecular formula of the expressed protein was C₃₀₁₃H₅₀₂₃N₁₀₀₅O₁₂₆₀S₁₉₉,the molecular weight was 81.867 94 ku,the theoretical isoelectric point(pI)was 5.09,and the total number of atoms was 10 500.The expressed protein contained a typical WRKYGQK conserved domain,and the zinc finger structure was CX₅CX₂₃HXH,belonging to the second Ⅱ d subfamily.The secondary structural elements were α-helix,extended chain,β-folding and random coiling,among which the proportion of random curl was the highest,up to 61.68%.There were 29 phosphorylation sites in total,which might be located in the nucleus.There were cis-regulatory elements upstream of the promoter that might related to resistance stress response and cis-regulatory elements related to growth and development and hormone response.The gene was closely related to potato StWRKY5 gene,and the amino acid homology of the coding protein reached 95%.

A multiplex RT-PCR detection system was established and studied to target four sweet potato RNA viruses,Sweet potato feathery mottle virus (SPFMV),Sweet potato chlorotic stunt virus (SPCSV), Sweet potato virus G (SPVG)and Sweet potato virus C (SPVC),which caused serious damage in sweet potato production. Firstly,the specific primers for four viruses were designed by Premier 5.0 software and synthesized according to the conserved sequences of coat protein(CP)gene of these viruses from GenBank. The cDNA synthesized was used as a template,and 4 couples of selected primers were used simultaneously to amplify the target fragments by PCR. A best multiplex RT-PCR reaction system was tested by optimizing the annealing temperature,extension time,dNTPs amount,template amount,primer concentration and so on and an optimal quadruple RT-PCR system that could simultaneously detect these four sweet potato RNA viruses was established. The results showed that an accurate and sensitive multiplex RT-PCR method for detecting the four sweet potato viruses mentioned above with respective target band had been successfully established by challenge tests in which there would be no crossover among the virus primers. The multiplex RT-PCR method established in this study could detect four kinds of sweet potato virus at the same time. It was not only accurate and sensitive,but also reduced the cost of detection and improved the efficiency of virus detection.

The purpose was to prepare polyclonal antibody against the recombinant double CP of Potato leafroll virus (PLRV)and Potato virus S (PVS),and apply the polyclonal antibody to indirect ELISA and DAS-ELISA detections of PLRV and PVS.Prokaryotic expression vector pET22b-LRCP/SCP of the fused double CP gene of PLRV and PVS was constructed.After replacement of lysozyme treatment by ultrasonic disruption,inclusion body protein was extracted from the recombinant strain BL21(pET22b-LRCP/SCP),the target protein(recombinant double CP)was purified by nickel ion affinity chromatography and high-purity target protein of 51.2 ku was obtained.The high-purity recombinant double CP was used as antigen to immunize rabbits to prepare an antiserum with a titer of 1∶128 k.Specific reactions were respectively observed between the purified polyclonal antibody (IgG) against the recombinant double CP and the positive standard of PLRV or PVS and no cross-reaction was found between the purified IgG and other four potato major viruses (PVX, PVY, PVA and PVM). The purified IgG against the recombinant CP with the diluted concentration of 1∶3 200 still positively reacted with PLRV or PVS in indirect ELISA detection.The purified IgG and the alkaline phosphatase-conjugated IgG both with the diluted contraction of 1∶100 also positively reacted with PLRV or PVS positive standard in DAS-ELISA detection.The results showed that one type of the prepared IgG against the recombinant double CP could detect two viruses of PLRV and PVS by DAS-ELISA or indirect ELISA.

Small GTPases Rabs is a kind of monomer GTP binding protein,which can act as a molecular switch involve in variety of physiological and biochemical reactions in plant cells. In order to explore the relationship between potato small G protein StRab5b and anthocyanin synthesis,the StRab5b gene was overexpressed in potato by Agrobacterium mediated genetic transformation,try to study the effects of StRab5b gene on anthocyanin content and PAL activity. The result showed that overexpression of StRab5b gene significantly increased the anthocyanin content in potato leaves,stems and tubers. Compared with the control,the anthocyanin content in leaves of transgenic potatoes StRab5b-L2,StRab5b-L3 and StRab5b-L5 increased by 42%,24% and 54%,respectively;in stems was increased by 59%,37% and 32%,respectively;in tubers was increased by 118%,82% and 105%,respectively. At the same time,StRab5b gene also promoted the PAL activity,a key enzyme of plant anthocyanin synthesis. Compared with the control,the PAL activity in leaves of transgenic potatoes StRab5b-L2,StRab5b-L3 and StRab5b-L5 increased by 88%,63% and 66%,respectively;in stems increased by 48%,38% and 31%,respectively;in tubers increased by 98%,78% and 64%,respectively. In summary,small GTPases StRab5b increased the content of anthocyanin in potato by regulating PAL activity.

In this study,the transcription level of resistance genes to the infection of Colletotrichum gloeosporioides were explored at the molecular level and provided candidate genes for further study of disease resistance mechanism.Suyu 8 is a new yam variety highly resistant to anthracnose,which was bred by tissue culture mutagenesis of the highly susceptible strain 024.In order to find out the anthracnose resistance genes,Suyu 8 and strain 024 leaves were inoculated for different time by Colletotrichum gloeosporioides strain 4-2,the differentially expressed genes were analyzed by transcriptome sequencing.Concurrently,the uninoculation leaves of Suyu 8 were used as control.The number of differentially expressed genes,inoculation for 24,48,72,96 h,were 197,132,187 and 313,respectively.After removing the common differentially expressed genes at each time point,we obtained 711 differentially expressed genes.Go enrichment showed that the differential genes were mainly related to response to biological stimulation,defense response,cell wall metabolism and oxidation-reduction process.KEGG enrichment analysis revealed that many metabolic pathways related to plant disease resistance were changed.The expression of multiple genes varied which defense-related plant hormone signal transduction pathways,such as auxin,jasmonic acid,and ethylene.Among them,five early auxin-responsive genes,the key genes of jasmonic acid and abscisic acid biosynthesis were up-regulated,while ERF036 was down-regulated,which might negatively regulate the infection of Colletotrichum gloeosporioides.Several cytochrome P450 genes,ubiquitin ligases involved secondary metabolite modification and phytosterol synthetases,defensins and lectins were up-regulated.WRKY,MYB and TIFY transcription factors positively or negatively regulate the expression of disease resistance genes.Under the regulation of transcription factors,PR protein,NBS-LRR disease resistance genes,receptor kinases were highly expressed,the expressions of CAT and SOD genes of antioxidant protective enzyme systems were activated by reactive oxygen species.In addition,the enzymes related to starch and sucrose synthesis were up-regulated,and enzymes related to starch degradation were down regulated.The expression trends of LAX4,IAA4,IAA21 were consistent with that of transcriptome sequencing.The relative expression of LAX4,IAA4 and IAA21 in resistant varieties were significantly higher than susceptible varieties,suggesting that auxin signaling pathway is beneficial to the immune response to anthracnose for yam.

The aim of this research was to clone IPT(Isopentenyl transferase,IPT ) genes in yam,predict the structure and property of DoIPT proteins and identify the expression of DoIPTs during bulbil sprouting.DoIPTs were cloned by homologous cloning techniques.The structure and property of DoIPT proteins were predicted by bioinformatics analysis.Real-time PCR technique was applied to identify the expression of these genes.Three nucleic acid sequences of 1 398,1 044,1 349 bp were obtained from yam.The three sequences were named DoIPT1,DoIPT5a and DoIPT5b,and the registration numbers of these genes were MW353160,MW353016 and MW353017,respectively.The reading frame lengths were 1 137,861,1 011 bp respectively,and the encoded amino acid lengths were 378,286,336 aa,respectively.Bioinformatics analysis showed that DoIPTs were unstable,hydrophilic and external family proteins.There were two signal peptides in DoIPT5b,and no signal peptide was found in the other two proteins.The secondary structures in the three proteins were consistent with the prediction models of the tertiary structures of three genes.The encoded regions of DoIPTs had high similarity with the isopentenyl transferase protein of other plants.Coding areas of DoIPTs showed that the N-terminal regions of DoIPTs contained the P-loop NTPase structural domain GXXGXGKS(T),which was able to catalyse cytokinin synthesis.The results of Real-time PCR technique showed that the expression of DoIPT5a in sprouting bulbils was highest under the condition of water shortage at 22 ℃;and the expression of DoIPT1 and DoIPT5b genes was the highest in a humid environment;the expression of DOIPTs genes raised with paclobutrazol treatment.These results laid a foundation for further study on the function of DoIPTs in yam.

In order to explore the effects of different concentrations of exogenous melatonin on the growth of potato seedlings under low temperature stress after cold acclimation,using Yunnan main potato variety Hezuo 88 as material,the effects of different concentrations of exogenous melatonin(50,100,150 μmol/L)on the growth and active oxygen metabolism system of potato seedlings under low temperature stress(4 ℃ 14 d ,-2 ℃ 12 h)were studied.The purpose of this study was to explore the feasibility of exogenous MT in alleviating the injury of potato seedlings caused by low temperature stress.The results showed that compared with potato seedlings under low temperature stress,the survival rate of seedlings treated with 50 μmol/L MT increased by 38.89 percentage point,and the plant height and stem diameter increased by 2.92% and 1.86%,the contents of SS,SP and Pro increased by 12.66%,13.80% and 1.96%,the activities of POD and CAT increased by 29.24% and 351.62%.The plant height and stem diameter of seedlings treated with 100 μmol/L MT increased by 16.97% and 2.82%,the contents of MDA and H₂O₂ decreased by 19.35% and 0.48%,the activities of POD and CAT increased by 55.44% and 213.56%.The survival rate of seedlings treated with 150 μmol/L MT increased by 27.78 percentage point,the plant height and stem diameter increased by 5.62% and 13.20%,the content of MDA decreased by 9.45%,the content of Pro increased by 38.70%,SOD,POD and CAT activity increased by 52.61%,25.25% and 300.94%,and the content of AsA and GSH increased by 3.48% and 3.97%.The results showed that exogenous MT could promote the growth of potato seedlings,reduce the content of MDA and H₂O₂ in leaves,promote the accumulation of SS,SP and Pro,increase the activities of SOD,POD and CAT,and increase the content of AsA and GSH.Thus,it can alleviate the damage of low temperature stress to potato seedlings,and then improve the ability of potato seedlings to resist low temperature stress.In summary,50 μmol/L MT treatment could better improve the stress resistance of potato seedlings under low temperature stress.

This article aims to explore the control methods of sweetpotato storage and provide a theoretical basis for regulating sweetpotato storage.Pre-production regulation is an important part of sweetpotato storage regulation, and fertilization is one of the important means of pre-production regulation. This study starts with fertilizer and uses two varieties of Shangshu 19 and Xinxiang as experimental materials. Nitrogen and phosphate fertilizers, and applying different gradients of potassium fertilizer. After uniform harvesting, select sweetpotatoes with the same size and no damage, and store them in a cold storage at 14℃ and 85% humidity. Samples were taken during the storage period of 0, 15, 30, 60, 90 days, and the decay rate, cell wall composition and enzyme activity were measured respectively, so as to explore the effect of different potassium application on the decay rate, cell wall composition and degrading enzymes of sweetpotato. The results of the study showed that the sweetpotatoes with different potassium application rates showed different degrees of decay during storage. Among them, the potassium application rate of Shangshu 19 was 150 kg/ha(Shangshu 19 K₂)and the potassium application rate of Xinxiang was 225 kg/ha(Xinxiang K₃), which had better storability and the lowest rot rate. Respectively, 18.25% and 23.00%, and Shangshu 19 K₂ had a higher cellulose content, which was as high as 80.63 mg/g, the original pectin content of Xinxiang K₃ is significantly higher than other treatments, with an average of up to 1.14%, and β-Gal enzyme activity is significantly lower than other treatments. Applying potassium treatment to sweetpotato can slow down the degradation of sweetpotato cell wall components and improve the ability to maintain the integrity of the cell wall, thereby reducing the rate of decay.

In order to reveal the response relationship between potato yield and rhizosphere soil microbiological characteristics under plastic film mulching and supplementary irrigation technology in semi-arid area,field experiments of potato mulching and supplementary irrigation were carried out in Dingxi irrigation experimental station of Gansu Province,which included six treatments,drip irrigation under plastic film(PFID),drip irrigation under semi mulching(PHID),furrow irrigation with full film mulching(PFIF),furrow irrigation without film mulching(PNIF),border irrigation with plastic film mulching(PFIB),and no irrigation in border field covered with plastic film(PFIBN).Flat planting without film and no irrigation(PNIN) was used as control.The effects of different treatments on potato yield,rhizosphere soil organic matter(TOM),organic carbon(TOC),microbial biomass carbon(MBC),microbial biomass nitrogen(MBN),urease and catalase were analyzed.The results showed that mulching methods had significant effects on rhizosphere soil microbiological properties under drip irrigation and furrow irrigation.MBC and MBN of full film mulching treatment were significantly higher than that of half mulching treatment,while urease and catalase activities of semi mulching or no mulching treatment were significantly higher than that of full mulching treatment(P <0.05).On the other hand,the activity of urease and catalase of all supplementary irrigation treatments(PFID,PHID,PFIF,PNIF,PFIB) were significantly higher than control treatment(PNIN).The potato yield,number of tubers per plant and tuber weight per plant of supplemental irrigation treatment were also significantly higher than control treatment.In particular,the yield of semi mulched drip irrigation treatment(PHID) was 44 603.70 kg/ha,which was 51.00% higher than control.Correlation analysis showed that potato yield was highly significant correlated with MBC,urease activity and catalase(P <0.01),and positively correlated with TOM and TOC(P <0.05).

In order to clarify the diversity and differences of endophytic bacteria in leaves of different potato varieties and provide basis for studying the mechanism of potato stress tolerance and the development of endophytic bacteria.The endophytic bacteria from the leaves of four main potato varieties in Yunnan Province were isolated and cultured on LB medium,all those strains were identified based on the morphological characteristics and the 16S rDNA sequence.There were 10 species of endophytic bacteria belonging to 9 genera,8 families and 3 phyla were isolated from Qingshu No.9 leaves. Staphylococcus and Exiguobacterium were the dominant genera,and the relative abundance of both genera were 18.18%.There were 12 species of endophytic bacteria belonging to 11 genera,11 families and 4 phyla were isolated from Hui-2 leaves. Acinetobacter and Pseudomonas were the dominant genera,and the relative abundance of both genera were 15.38%.There were 13 species of endophytic bacteria belonging to 10 genera,9 families and 4 phyla were isolated from Hezuo No.88 leaves. Acinetobacter were the dominant genera,with a relative abundance of 23.53%.There were 17 species of endophytic bacteria belonging to 8 genera,7 families and 4 phyla were isolated from Lishu No.6 leaves. Staphylococcus were the dominant genera,with a relative abundance of 22.22%.There were 37 species of culturable bacteria were isolated from the leaves of four potato varieties.There were 4,8,4,10 unique endophytic bacteria from the leaves of Qingshu No.9,Hui-2,Hezuo No.88 and Lishu No.6,respectively.The species composition and dominant genera of endophytic bacteria in leaves of different potato varieties were different,the results would provide bacteria resources for studying the function of endophytic bacteria in potato leaves.

In order to study the molecular mechanism of purple sweet potato leaf in response to enhanced UV-B radiation, the leaf of Dianziganshu 24(DZS24) was used as the test material and transcriptome sequencing analysis was performed by high-throughput sequencing technology under natural light and artificial supplementary the UV-B(T=7.2 kJ/(m² · d)). The results showed that the expression of 477 genes(DEGs) were changed significantly, among which 382 and 95 were up-and down-regulated ones. GO analysis showed that these DEGs were mainly significantly enriched in oxidation-reduction process of biological process and oxidoreductase activity of molecular function. KEGG pathway analysis showed that a total of 9 pathways were significantly enriched, among which the biosynthesis pathway of secondary metabolites contained the largest number of DEGs. The DEGs of purple sweet potato leaf in response to enhanced the UV-B were mainly CHS (Tai6.1732, Tai6.53503, Tai6.45381) and DFR (Tai6.3019) in the flavonoid synthesis pathway as well as the CYP83B1 (Tai6.44115, Tai6.18064, Tai6.41206) in the glucosinolate biosynthesis and CYP85A1 (Tai6.2806, Tai6.19253, Tai6.15801) in brassinosteroid biosynthesis. The main transcription factors were C2H2, NAM, bHLH and RR-A-type. In summary, the key genes might be IbCHS, IbDFR, IbCYP83B1 and IbCYP85A1, as well as the main transcription factors might be C2H2, NAM, bHLH and RR-A-type in the purple sweet potato leaf response to the UV-B enhancement.

Enhanced UV-B radiation can cause damage to plants, and melatonin can enhance plant resistance. At present, there is no research report on the effect of melatonin on potatoes under UV-B radiation, so this experiment measured plant height, photosynthetic parameters(Pn, Gs, Ci, and Tr), fluorescence parameters(Fv/Fm, ETR, qP, and NPQ), 1, 5-bisphosphate ribulose carboxylase(Rubisco) and phosphoenolpyruvate carboxylase(PEPC), to analyze the effect of melatonin on the photosynthetic and fluorescence characteristics of potatoes under UV-B radiation, provide scientific basis for revealing the defense mechanism of melatonin against potato plants under UV-B radiation. The potato variety Hezuo 88 planted in greenhouse was used as experimental material. The concentrations of melatonin were set at 0, 25, 50, 100, 150, 200 μmol/L respectively under natural light(CK) and three enhanced UV-B radiation((2.5, 5.0, 7.5 KJ/(m²·d)).The plant height, Rubisco, PEPC, photosynthetic parameters and fluorescence parameters (except NPQ) of potato decreased after exposure to enhanced UV-B radiation, but increased after exposure to melatonin.Melatonin can improve the resistance of potato to UV-B radiation, increase the plant height and enhance its photosynthetic capacity. However, the trend of these indicators rising first and then decreasing also indicates that excessive melatonin concentration will also cause certain damage to plants.

In order to obtain potato plants which could be resistant to multiple viruses (viroid) simultaneously, P25, HC-Pro, and Virp1 gene targeting PVX, PVY, and PSTVd proteins were cloned, respectively. Three types of amiRNAs targeting sequences encoding the silencing suppressor P25, HC-Pro and Virp1 were designed by using Arabidopsis thaliana miR159a as backbone. These three amiRNA sequences were connected by Overlapping PCR. The synthetic of P25, HC-Pro and Virp1 gene was inserted into the expression vector pCAMBIA1300-221 to form p1300-221-preamiR-P25-HCPro-Virp1, and the vector was verified by PCR and restriction enzyme digestion. Pre-amiR-P25-HCPro-Virp1 was transformed into minituber of potato cultivar Favorita by Agrobacterium tumefaciens infection. 15 transformed plants were obtained through regenerating, pressure screening and differentiation.PCR results showed that 10 of them were detected the same size fragment (729 bp) as the target gene. Further qRT-PCR testing confirmed that amiR-P25-HCPro-Virp1 gene was expressed in 10 transformed plants, and the relative expression was between 7.68-21.37. Inoculated the T₀ transgenic plants with the mixture virus of PVX, PVY and PSTVd by friction into the leaves of the plants at the 6-8 leaf stage. The plant growth was observed after 20 days and the results showed that the control plants were severely susceptible, with symptoms such as short plants and mottled leaves, while the transformed plants did not show infection symptoms, and growth normally. There was no virus detected by RT-PCR either. This indicated that the transfected amiR-P25-HCPro-Virp could be stably expressed in the transformed plants, and the transformed plants were resistant to PVX, PVY and PVSTd viruses (viroid). Transformed plants resistant to PVX, PVY and PVSTd were obtained, and the resistance was significant, which provided new genetic resources for potato virus-resistant breeding.

In order to investigate the function of StTCP13 in salt stress and lay the foundation for the functional study of StTCP13 in response to abiotic stress of potato. The coding sequence of StTCP13 was obtained by homologous cloning. The protein domain and phylogenetic tree were analyzed by bioinformatics.The results showed that the StTCP13 gene was cloned from the potato variety Favorita.The coding sequence length of StTCP13 was 669 bp, encoding a protein composed of 222 amino acid residues.The results of prokaryotic expression suggested that the molecular weight of fusion protein was 51 ku.Protein domain analysis suggested that the protein contained a conserved TCP domain.The results of phylogenetic tree revealed that the protein had a high homology with the TCP13 in Solanum pennellii and Capsicum annuum.Expression pattern of StTCP13 was analyzed by qRT-PCR.Expression analysis showed that the StTCP13 exhibited different expression levels in root, stem, leaf, bud and bud eye in potato. The StTCP13 had high expression level in root, low expression level in bud eye, in addition,the FPKM of StTCP13 in Spud DB showed that the expression of StTCP13 was induced by high-salinity, drought and ABA treatments.The StTCP13-pGEX6P1 prokaryotic expression vector was constructed by double enzyme digestion. Prokaryotic expression results showed that the StTCP13-GST tag fusion protein was successfully expressed in Escherichia coli BL21 . Salt stress phenotype analysis suggested that StTCP13 could improve the tolerance to salt stress in E.coli BL21 . Above all, the StTCP13 might play a role in response to salt stress in potato.

In order to determine the difference of molecular and cellular genetics of six new potato strains (NNS-P 11, NNS-H 6,NNS-H 4, NNS-C 10, NNS-L 9 and NNS-Y 40), with MIN-021 as control material, conventional compression and SSR molecular labeling were used to compare and analyze the traits of the six new strains, include pollen fertility, PMCMⅠ chromosome pairing behavior, SSR fingerprint characteristics. This would provide the basis for further breeding, identification and registration of new potato varieties. The results showed that:There were some differences in pollen fertility among six new potato strains. The new strain NNS-Y 40 had the lowest pollen fertility (39.95%), the new strains NNS-P 11, NNS-H 4, NNS-C 10 had the highest pollen fertility (81.12%, 77.94%, 77.76%), the new strains NNS-H 6 and NNS-L 9 pollen fertility in the middle of the lowest and highest (67.27%, 69.71%). The chromosome pairing configuration of six new strains NNS-P 11, NNS-H 6, NNS-H 4, NNS-C 10, NNS-L 9 and NNS-Y 40 were 1.08Ⅰ+14.89Ⅱ+1.18Ⅲ+3.4 0Ⅳ, 2.35Ⅰ+11.96Ⅱ+3.59Ⅲ+2.74Ⅳ, 2.10Ⅰ+14.21Ⅱ+1.35Ⅲ+3.02Ⅳ, 3.31Ⅰ+13.98Ⅱ+2.99Ⅲ+1.94Ⅳ, 3.45Ⅰ+12.94Ⅱ+2.81Ⅲ+2.56Ⅳ and 8.23Ⅰ+6.92Ⅱ+6.07Ⅲ+1.93Ⅳ. The genomic DNA of six potato strains and control sample was amplified with 10 pairs SSR selected primers and 82 polymorphic bands were obtained, with the polymorphic ratio was 78.10%. The genetic distance (GD) of new potato strains ranged from 0.244 4-0.687 5. Base on 0.40 genetic distance, seven test materials were divided into three categories:the first category was new strains NNS-P 11, NNS-L 9, NNS-Y 40 and MIN-021, the second category was new strain NNS-H 6 and NNS-H 4, and the third category was new strain NNS-C 10. In addition, a SSR fingerprint was established to distinguish 7 test materials.

In order to explore the function of StWRKY57 (PGSC0003DMT400072958) in potato, a full-length cDNA sequence of the homologous StWRKY57 gene in Solanum tuberosum PB06 named StWRKY57-PB was cloned by RT-PCR and bioinformatics methods were used to analyze the sequence characteristic of StWRKY57-PB. For prokaryotic expression and purify, the prokaryotic expression vector of StWRKY57-PB was constructed based on pGEX-KG, then fusion protein was induced in Escherichia coli strains. The results showed that the StWRKY57-PB gene contained a 981 bp open reading frame(ORF), and the coding protein belonged to the Ⅱ c class of the WRKY transcription factor family, containing 326 amino acid residues with a WRKY(WRKYGQK) domain and a zinc-finger-like motif(C-X₄-C-X₂₃-H-X₁-H). StWRKY57-PB was determined structurally to be 16.56% a-helix, 13.50% extended strand, 7.98% β-turn and 61.96% random coil. Compared with StWRKY57(XP_006348711.1), there were 4 amino acid mutations in StWRKY57-PB. Multiple alignment and phylogenetic tree analyses indicated that StWRKY57-PB showed the next-closest relationship with SlWRKY57 with 93% similarity and was farthest to AtWRKY57. StWRKY57-PB protein was stably expressed in three Escherichia coli strains of Rosetta, BL21 and Tuner with 0.5 mmol/L IPTG. Fusion protein of GST-StWRKY57-PB was purified through GST SefiroseTM resin, and then Western Blot was carried to verify the successful expression and purification of the protein. It is of great significance to further determine the activity and biological function of this protein in vitro.

In order to reveal the genetic diversity of sweet potato and its wild species and analyze the genetic relationship between sweet potato and its wild species, SCoT molecular markers were used to analyze the results of PCR amplification, genetic diversity, evolutionary relationship and population structure of 22 germplasm of sweetpotato and its wild species I.trifida and I.triloba in the present study. The results showed that 278 stable bands were detected by 43 primers, 4-12 bands could be detected by each primer, and five primers with rich polymorphism were screened, which were SCoT-8, SCoT-20, SCoT-26, SCoT-36 and SCoT-35. These primers could be used in the study of sweet potato molecular markers in the future. The analysis of genetic diversity based on the results of molecular markers showed that the highest genetic diversity was in I.trifida, followed by I.batatas, and the lowest in I.triloba. According to the results of genetic cluster analysis and population genetic structure analysis, 22 germplasm were divided into three groups, and the classification of each group was basically the same as its species. There was less blood exchange between different species, and the genetic composition was relatively simple. In comparison, the blood exchange between sweet potato and I.trifida was more than that between I.triloba. In conclusion, the results of this study preliminarily proved the potential application of SCoT molecular markers in the study of sweet potato evolution, and revealed the different genetic relationships between sweet potato and its wild relatives, which provided theoretical support for introducing I. trifida and other wild species into sweet potato breeding to improve the genetic diversity of sweet potato in the future.

In order to further clarify the spatial and temporal accumulation characteristics of cadmium in sweetpotato and to explore the causes of low cadmium accumulation in fleshed storage root,the temporal and spatial differences of cadmium accumulation in Xiangshu 20 were detected by atomic absorption spectrometry(AAS)in this study,and the expression difference of homologous genes related to cadmium accumulation in sweetpotato were analyzed by Fluorescence quantitative PCR.The results showed that the cadmium accumulation of sweetpotato followed the characteristics of root>stem>leaf. When plants were growth in the soil with 0.6 mg/kg cadmium,the increase rate of cadmium concentration in sweetpotato range from 3.05 to 8.75,and when plants growth in the soil with 2.0 mg/kg cadmium,the increase rate of cadmium concentration in sweetpotato range from 6.25 to 26.89. The accumulation rate of cadmium in shoots and leaves was higher than that in fleshed storage roots. The cadmium accumulated in leaves was not easy to transfer again,so cadmium concentration in new leaves was lower than that in old leaves. The homologous genes related to plant cadmium accumulation and transport existed in sweetpotato genome,and most of them had multiple homologous copies. Although these genes were induced to express with the increase of cadmium accumulation in plants,the spatiotemporal expression patterns of these genes were different. Compared with the expression pattern of genes related to cadmium accumulation,the expression of cadmium stress response related genes was been induced by cadmium accumulation more obvious. The cadmium accumulation in sweetpotato increased with the increase of soil cadmium concentration,and increased with the extension of growth time. The accumulation of cadmium in fleshed storage roots was lower than that in peels,stems and leaves,and the accumulation rate of cadmium in fleshed storage roots were much lower than that in shoots with the extension of growth time. The homologous genes related to plant cadmium accumulation and transport existed in sweetpotato genome,and most of them had multiple homologous copies. The spatiotemporal expression patterns of these genes were different,it was suggested that this expression difference was related to the cadmium accumulation characteristics of sweetpotato.