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    Effects of Exogenous Hormones on Growth,Yield and Physiological Characteristics of Tigernut(Cyperus esculentus L.)under Drought Stress
    LIU Jiayao, CHENG Yan, WEI Zunmiao, LONG Wei, WANG Liang, MU Zhongsheng
    Journal of Henan Agricultural Sciences    2022, 51 (11): 42-49.   DOI: 10.15933/j.cnki.1004-3268.2022.11.005
    Abstract (1938)      PDF (7617KB)(108)       Save
    In order to determine the optimal spraying concentration of exogenous hormones on tigernut under drought stress,Jisha No.2 was used as material,drought stress was simulated by 15% polyethylene glycol(PEG 6000)to study the effects of different concentrations of gibberellin(GA3,0.1,0.2,0.3,0.4,0.5 μg/L),melatonin(MT,20,40,60,80,100 μmol/L)and salicylic acid(SA,0.25,0.50,0.75,1.00,1.25 mmol/L) on the growth,yield,relative conductivity,chlorophyll content,proline content,malondialdehyde(MDA)content,superoxide dismutase(SOD)and peroxidase(POD)activities of tigernut seedlings.The results showed that under drought stress,spraying the three kinds of exogenous hormones could promote the growth and yield of tigernut,G3(0.3 μg/L),M3(60 μmol/L),S2(0.50 mmol/L)treatments had the best effect,the yields of which significantly increased by 38. 32%,31.73% and35.90% compared with the drought stress treatment(CK2)respectively. Under drought stress,spraying the three kinds of exogenous hormones could reduce the relative conductivity and MDA content in leaves,and increase the chlorophyll content,proline content,SOD and POD activities,alleviating the damage caused by drought stress to the seedlings of tigernut,the best treatments were G3,M3 and S2,respectively.Compared with CK2,for G3,M3 and S2 treatments,the relative conductivities significantly decreased by 54.92%,34.44% and 27.86%,chlorophyll contents obviously increased by 33.83%,19.03% and 15.84%,proline contents significantly increased by 478.88%,627.62% and 412.87%,MDA contents significantly decreased by 53.51%,53.43% and 54.75%,SOD activities increased by 0.95%,13.51%and 16.25%,POD activities increased by 1.80%,10.36% and 12.84%,respectively.In conclusion,under drought stress,spraying appropriate concentrations of GA3,MT and SA could promote the growth of tigernut,alleviate the damage caused by drought stress,and then increase the yield,overall the best concentrations of GA3,MT and SA were 0.3 μg/L,60 μmol/L and 0.50 mmol/L,respectively.

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    Effect of Colchicine Mutagenesis on Emergence Rate and Quality of Tigernut(Cyperus esculentus L.)
    WEI Zunmiao, LIU Jiayao, CHENG Yan, WANG Zhanhai, XU Fangda, CHEN Xin, MU Zhongsheng
    Journal of Henan Agricultural Sciences    2022, 51 (11): 50-55.   DOI: 10.15933/j.cnki.1004-3268.2022.11.006
    Abstract (1613)      PDF (5164KB)(100)       Save
    In order to determine the half‑lethal dose for colchicine mutagenesis of tigernut(Cyperus esculentus L.)tuber and obtain high quality variant germplasm resources,the tubers before and after germination were mutagenized with different concentrations(0,1,2,3,4 g/L)of colchicine,the emergence rate was calculated one month later,and the quality was tested by the near‑infrared(NIR)grain analyzer after maturity.The results showed that whether mutagenesis was done before or after germination,the emergence rate of tigernut decreased with the increase of colchicine concentration and mutagenesis time,colchicine concentration had extremely significant effect on the emergence rate,mutagenesis time had significant effect on the emergence rate.Before germination,tigernut tuber was more sensitive to colchicine,the half‑lethal dose of colchicine was lower than that of mutagenesis after germination,and mutagenesis before germination could cause bud deformity,but buds of mutagenesis treatment after germination did not obviously change.On the principle of low mutagenesis concentration and short mutagenesis time to half‑lethal dose,the optimal half‑lethal dose of colchicine was colchicine concentration of 1 g/L,mutagenesis for 24 hours before germination of tigernut tuber.By testing,one germplasm resource with starch content higher than 32% and six germplasm resources with total sugar content higher than 36% in tuber were obtained,which layed the foundation for high‑quality breeding.

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    Genome Size,Ploidy and Phylogeny of Cyperus esculentus L.
    WANG Huiwei, ZHU Shixin, ZHANG Xinyou, WANG Yan, YANG Tiegang, ZHANG Xiangge, WANG Shufeng, LI Chunxin
    Journal of Henan Agricultural Sciences    2023, 52 (1): 34-41.   DOI: 10.15933/j.cnki.1004-3268.2023.01.004
    Abstract (2023)      PDF (2024KB)(881)       Save
    In order to clarify the genome characteristics of the main varieties and characteristic germplasm of Cyperus esculentus L. in the Huang Huai region and the phylogenetic relationship between them and their related species,the genome size and ploidy of six C. esculentus L. materials were evaluated by flow cytometry and genome survey analysis,and the phylogenetic relationship between them and their related groups was analyzed based on the sequences of the external transcribed spacer(ETS) and internal transcribed spacer(ITS) of ribosomal DNA. The results of flow cytometry analysis showed that the genomes of six C.esculentus L. materials were 0.808 6—0.858 5 Gb,with an average of 0.826 4 Gb;the results of genome survey analysis showed that the genome size,proportion of repetitive sequences,GC content and heterozygous ratio of the three tuber types of C.esculentus L. were 0.697 9 Gb,81.02%,34.7% and 0.28% for Yuyousha 2,0.778 7 Gb,84.45%,36.4% and 0.08% for Yuyousha 3.and 0.790 6 Gb,83.75%,34.9% and 0.19% for YYS‑4,respectively;genome survey analysis and pollen staining results showed that the three tuber types of C.esculentus L. were triploid. Phylogenetic analysis showed that six C. esculentus L. were clustered into C.esculentus L.branch,this branch was closely related to the branch composed of C.rotundus L. and C.glomeratus L.,they formed sister group,and formed C4 plant branch with C.iria L. ,C.glomeratus L. and C.cuspidatus H.B.K.

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    Tissue Screening and Condition Optimization of Protoplast Preparation of Cyperus esculentus
    ZHANG Xiangge, ZHU Yajing, LU Lili, SONG Wanxian, ZHANG Xinyou, HU Haiyan, YU Meiqin, LI Juzheng, WANG Huiwei
    Journal of Henan Agricultural Sciences    2023, 52 (12): 49-56.   DOI: 10.15933/j.cnki.1004-3268.2023.12.006
    Abstract (1504)      PDF (2734KB)(240)       Save
    In order to obtain Cyperus esculentus protoplasts efficiently,the tissue parts including young shoots,young roots,young leaves,young leaf sheaths,tillering nodes and stolons were screened for protoplast preparation by single factor test method,and the enzymatic hydrolysis conditions such as enzyme combination dosage,enzymatic hydrolysis time,osmotic pressure stabilizer dosage and pH value of the enzyme solution were also optimized.The results showed that the best tissue part for protoplast preparation of Cyperus esculentus was young tillering node;and the optimal enzymatic hydrolysis conditions were as follows:the enzyme combination dosage was 2.0% cellulase+0.9% macerozyme,the enzymatic hydrolysis time was 6 h,the osmotic pressure stabilizer(mannitol)dosage was 11%,and the pH value of the enzyme solution was 6.0.Under these conditions,0.2 g of Cyperus esculentus tillering nodes were added to 4 mL of enzyme solution,and then the protoplast yield could reach 22.91×105 per gram and the protoplast activity could reach 90.72%.In this study,an efficient method for protoplast preparation of Cyperus esculentus was obtained,which could lay a foundation for subsequent protoplast fusion,germplasm innovation and hybrid breeding of Cyperus esculentus.
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    Metabolites Mining and Path Enrichment Analysis of Tigernut Tuber Formation
    GUO Xiaoyang, LA Guixiao, XU Xinran, YU Yange, DAI Dandan, LI Yanpeng, WANG Yanhong, GUO Hongxia, YANG Tiegang
    Journal of Henan Agricultural Sciences    2024, 53 (8): 30-43.   DOI: 10.15933/j.cnki.1004-3268.2024.08.004
    Abstract (1327)      PDF (4749KB)(1019)       Save
    In order to dig out the key metabolites in the process of forming tubers by the stolons of tigernut and regulate the stolons to form more tubers,the stolons and new tubers were taken as the research objects,and untargeted metabolomics analysis was carried out by high performance liquid chromatography Abstract:In order to dig out the key metabolites in the process of forming tubers by the stolons of tigernut and regulate the stolons to form more tubers,the stolons and new tubers were taken as the research objects,and untargeted metabolomics analysis was carried out by high performance liquid chromatography combined with triple quadrupole mass spectrometer.And then,the differential metabolites related to the formation of tubers were screened by multivariate statistical method,and KEGG metabolic pathways associated with differential metabolites were enriched.The results showed that a total of 1 217 metabolites were detected in unexpanded stolons(PFJ1),expanded stolons(PFJ2)and new tubers(JD1).According to the conditions of VIP(Variable importance in the projection)>1,P<0.05 and FC(Fold change)>2 or FC<0.5,35 differential metabolites were screened for JD1 vs PFJ1,of which 26 were up‑regulated and nine were down‑regulated;JD1 vs PFJ2 identified 57 differential metabolites,of which 50 were up‑regulated and seven were down‑regulated;PFJ2 vs PFJ1 identified 39 differential metabolites,of which three were up‑regulated and 36 down‑regulated. The metabolites were identified as amino acids and their derivatives,nucleotides and their derivatives,organic oxygen compounds,flavonoids,phenylpropanes,phenols and so on. The relative content analysis of differential metabolites showed that the content of glutathione in JD1 was 14.021 times that in PFJ1 and 29.195 times that in PFJ2.The content of 3‑methy‑L‑histidine in JD1 was 10.463 times that in PFJ1 and 129.931 times that in PFJ2.The content of kaempferol in JD1 was 0.421 times that in PFJ1 and 0.358 times that in PFJ2.KEGG pathway analysis of differentially expressed metabolites was further conducted,and they were mainly concentrated into KEGG pathways such as aminoacyl‑tRNA biosynthesis,arginine and proline metabolism,alanine,aspartic acid and glutamate metabolism,phenylalanine metabolism,flavonoid and flavonol biosynthesis.In summary,amino acids and flavonoids were the main metabolites related to the formation of tubers,and the metabolic pathways were mainly concentrated in aminoacyl‑tRNA biosynthesis and flavonoid and flavonol biosynthesis.

     
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    Establishment of Callus Induction and Regeneration System of Cyperus esculentus
    ZHU Yajing, ZHANG Xiangge, XU Xinran, HU Haiyan, CHENG Shan, CHEN Chen, LI Chunxin, SONG Wanxian, SONG Wenxu, WANG Huiwei
    Journal of Henan Agricultural Sciences    2024, 53 (10): 73-81.   DOI: 10.15933/j.cnki.1004-3268.2024.10.009
    Abstract (1935)      PDF (3147KB)(745)       Save
    In order to establish an efficient callus induction and regeneration system of Cyperus esculentus,the explants(young root,young leaf,young leaf sheath and tillering node)for callus induction were screened,and the effects of different plant growth regulator combinations on callus induction,proliferation and differentiation were explored.The results showed that the best explant for callus induction of Cyperus esculentus was tillering node;the optimal plant growth regulator combination for callus induction was 2.5 mg/L 2,4⁃D+0.6 mg/L 6⁃BA+0.6 mg/L NAA+0.9 mg/L ZT,and the induction rate could reach 65.58%;the optimal plant growth regulator combination for callus proliferation was 0.5 mg/L 2,4⁃D+0.9 mg/L 6⁃BA+1.2 mg/L NAA+1.2 mg/L ZT,and the proliferation rate could reach 68.12%;the optimal plant growth regulator combination for callus differentiation was 0.9 mg/L 6⁃BA+0.6 mg/L NAA,and the differentiation rate could reach 66.67%.In this study,the best explants suitable for callus induction of Cyperus esculentus were screened,and the conditions of callus induction,proliferation and differentiation of Cyperus esculentus were optimized,which could lay a foundation for the subsequent research and breeding application of biotechnology such as somatic hybridization and genetic transformation of Cyperus esculentus.
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