不同地面覆盖方式下新郁葡萄果皮黄酮转录组和代谢组联合分析

doi:10.6048/j.issn.1001-4330.2024.01.008

摘要/Abstract

摘要：

【目的】研究园艺地布覆盖和生草栽培两种不同地面覆盖方式下鲜食葡萄新郁果皮黄酮相关组分及其合成酶基因信息,为筛选新郁葡萄高效优质栽培新模式提供科学依据。【方法】以7年生新郁葡萄为试材,以清耕栽培为对照,分别进行行间铺设园艺地布和行间生草栽培两种地面覆盖方式处理,分析不同地面覆盖方式下葡萄果皮黄酮转录组和代谢物含量的差异。【结果】(1)园艺地布覆盖获得差异表达基因587个,其中上调基因317个、下调基因270个,生草栽培获得差异表达基因177个,其中上调基因56个、下调基因121个。(2)GO数据库将园艺地布覆盖注释的基因涉及45个功能组,将生草栽培覆盖注释的基因组涉及37个功能组,均主要集中在细胞组分、分子功能和生物学过程;KOG数据库分别将其注释到的175个和86个DEGs进行直系同源分类,各自获得22个和16个功能分类,园艺地布覆盖的193个差异基因被注释到KEGG通路中,与黄酮相关富集途径有次级代谢物生物合成、苯丙醇生物合成、类黄酮生物合成、苯丙氨酸生物合成、植物昼夜节律、二苯乙烯类二芳庚类和姜辣素类的生物合成;生草栽培的61个差异基因被注释到KEGG通路中,与黄酮相关富集途径有次级代谢物生物合成。(3)新郁葡萄果皮共检测到12类60个黄酮代谢物,其中地布覆盖差异代谢物7个(下调2个,上调5个),生草覆盖差异代谢物11个(全部下调)。黄烷醇含量占总黄酮含量的52.41%~63.70%,所占比例最高,生草栽培和园艺地布栽培的黄烷醇类含量均高于对照,分别比对照提高了19.42%和7.55%。(4)与黄酮合成相关富集途径主要集中在类黄酮生物合成、黄酮和黄酮醇生物合成,园艺地布覆盖筛选出7个候选基因:C4H、CHS、F3H、LAR、HCT、FLS和C12RT1,生草筛选出5个候选基因:CHS、F3H、F3'5'H、FLS和LAR。【结论】筛选出在地布和生草栽培模式下新郁葡萄黄酮生物合成候选基因。在新疆吐鲁番极端干旱地区鲜食葡萄新郁地面覆盖方式可选择生草栽培。

关键词: 新郁葡萄; 地面覆盖; 黄酮; 转录组; 代谢组

Abstract:

【Objective】 To clarify the flavonoid related components and synthase genes information of table grapes under two different ground covering methods of horticultural cloth covering and grass cultivation in the hope of laying the theoretical foundation for proposing a new mode of high efficiency and high quality cultivation of Xinyu grapes in production. 【Methods】 In this study,the ground mulched by black geotextile and grass,clean tillage was taken as the control,the cDNA of Xinyu grape peel flavonoids were sequenced and were analyzed by using the bioinformatics methods subsequently,such as sequencing assess and gene function annotation,at the same time,the flavonoid metabolites in grape peel were determined by LC-MS/MS. 【Results】 (1) Compared the transcriptome of the black geotextile mulch and grass,587 differentially expressed gene(DEGs) in black geotextile mulch were found,including 317 up-regulated gene and 270 down-regulated genes,177 differentially expressed gene(DEGs) in grass were found,including 56 up-regulated gene and 121 down-regulated genes.(2) GO function annotation gene of black geotextile mulch was divided into 45 function categories,and the GO annotation gene of grass type was divided into 37 function categories,in which many function categories were mainly involved,such as cellular component,molecular function and biological processes. KOG database classified the 175 and 86 DEGs annotated to,respectively,with 22 and 16 functional categories respectively,and 193 DEGs for black geotextile mulch were annotated into KEGG pathway,biosynthesis of secondary metabolites,phenylpropanoid biosynthesis,flavonoid biosynthesis,phenylalanine biosynthesis,circadian rhythm-plant,stilbenoid,diarylheptanoid and gingerol biosynthesis were associated with flavonoid enrichment pathway,61 DEGs for grass were annotated into KEGG pathway,biosynthesis of secondary metabolites was associated with flavonoid enrichment pathway.(3) A total of 60 flavonoids metabolites were detected for Xinyu grape peel and divided into 12 categories,including 7 differential metabolites(2 down-regulated and 5 up-regulated) for black geotextile mulch and 11 differential metabolites(all down-regulated) for grass cultivation. Flavanols proportion content was the highest,with 52.41%-63.70%,flavanols content were higher for grass cultivation and black geotextile mulch than for the control,with 19.42% and 7.55%.(4) The flavonoid biosynthesis related enrichment pathways for black geotextile mulch were mainly concentrated in the flavonoid biosynthesis,while the flavonoid biosynthesis related enrichment pathways of grass mulch were mainly concentrated in the secondary metabolites and flavonoid biosynthesis,seven candidate genes were screened including C4H,CHS,F3H,LAR,HCT,FLS and C12RT1 for black geotextile mulch,five candidate genes were screened including CHS,F3H,F3'5'H,FLS and LAR for grass. 【Conclusion】 By transcriptome and metabolite analysis,candidate genes for flavonoid biosynthesis of Xinyu grape were screened and analyzed for black geotextile and grass mulch,grass cultivation was the preferable cultivation method for Xinyu grape in Xinjiang Turpan extreme drought area.

Key words: Xinyu grape; ground mulch; flavonoids; transcriptome; metabolomics

中图分类号:

S662.1

户金鸽, 白世践, 陈光, 蔡军社. 不同地面覆盖方式下新郁葡萄果皮黄酮转录组和代谢组联合分析[J]. 新疆农业科学, 2024, 61(1): 63-78.

HU Jinge, BAI Shijian, CHEN Guang, CAI Junshe. Transcriptome and metabolome integrated analysis of flavonoids in Xinyu grape peel under different ground mulch 5ypes[J]. Xinjiang Agricultural Sciences, 2024, 61(1): 63-78.

图/表 12

表1 不同地面覆盖方式转录组测序质量变化

Tab.1 Quality analysis of transcriptome sequencing of different mulching type

样本ID Sample ID	原始读数 Raw reads	干净数据 Clean reads (bp)	碱基总数 Clean base (G)	错误率 Error rate (%)	测序准确度在99% Q20(%)	测序准确度在99% Q30 (%)	G和C 2种碱基占总碱基的百分比GC(%)
清耕Q-1 Clean tillage Q-1	57476100	55348990	8.30	0.03	97.00	91.94	46.03
清耕Q-2 Clean tillage Q-2	46791054	41704844	6.26	0.03	98.04	94.21	46.30
清耕Q-3 Clean tillage Q-3	47866130	41706034	6.26	0.03	98.01	94.16	46.10
园艺地布F-1 Horticultural ground cloth F-1	48420302	42003268	6.30	0.02	98.09	94.34	45.95
园艺地布F-2 Horticultural ground cloth F-2	48354116	44252400	6.64	0.03	97.89	93.91	46.46
园艺地布F-3 Horticultural ground cloth F-3	46448184	42513692	6.23	0.03	98.04	94.20	46.55
生草栽培S-1 Sod-culture-1	57847986	55154966	8.27	0.03	96.81	91.59	46.23
生草栽培S-2 Sod-culture-2	51485636	49924206	7.49	0.03	96.90	91.57	44.84
生草栽培S-3 Sod-culture-3	52146666	50057894	7.51	0.03	96.93	91.84	46.16

表2 差异基因数量

Tab.2 Statistics of the number of differential genes

组 Group	差异基因总数 Total number of differentially expressed genes	上调基因数 Upregulated gene count	下调基因数 Number of down- regulated genes
清耕vs园艺地布Q_vs_F Clean tillage vs Horticultural ground cloth Q_vs_F	587	270	317
清耕vs生草栽培Q_vs_S Clean tillage vs Sod-culture Q_vs_S	177	121	56

图1 不同地面覆盖方式下葡萄果皮差异基因二级条目分类注(Note):生物学过程(Biological process);细胞组分(Cellular component);分子功能(Molecular function);细胞膜(Membrane);胞外区(Extracellular region);细胞(Cell);细胞连接(Cell junction);膜内腔(Membrane-enclosed lumen);蛋白复合体(Protein-containing complex);细胞器(Organelle);细胞外基质部分(Extracellular region part);细胞器组件(Organelle part);细胞膜组件(Membrane part);细胞组件(Cell part);共质体(Symplast);超分子复合物(Supramolecular complex);翻译调节活性(Transcription regulator activity);催化活性(Catalytic activity);营养储层活动(Nutrient reservoir activity);结构分子活性(Structural molecule activity);转运器活动(Transporter activity);绑定(Binding);抗氧化活性(Antioxidant activity);分子功能调节(Moleular function regulator);分子传感器活性(Molecular transducer activity);生殖(Reproduction);代谢过程(Metabolic process);生长(Growth);细胞增殖(Cell proliferation);细胞进程(Cellular process);生殖过程(Reproductive process);生物黏附(Biological adhesion);信号(Signaling);多细胞有机体进程(Multicellular organismal process);发育进程(Developmental process);运动力(Locomotion);节律进程(Rhythmic process);生物过程的正调控(Positive regulation of biological process);生物过程的负调控(Negative regulation of biological process);应激反应(Response to stimulus);定位(Localization);多有机体进程(Multi-organism process);生物调节(Biological regulation);细胞成分组织或生物合成(Cellular component organization or biogenesis);解毒(Detoxification);横坐标表示二级GO条目,纵坐标表示GO条目的差异基因的数量(The abscissa represents the second-level GO entry,and the ordinate represents the number of differential genes in the GO entry)

Fig.1 Classification map of secondary entries of grape peel differential genes by different mulch type

表3 GO数据库三大功能主类DEG数

Tab.3 DEG number of three major GO database functions

差异表达基因集 Differentially expressed gene set	DEG数(个)Number of DEG			有GO功能注释的DEG数 Number of DEGs with GO function annotations	DEG总数 Total of DEG
差异表达基因集 Differentially expressed gene set	生物学过程 Biological process	细胞组分 Cellular component	分子功能 Molecular function	有GO功能注释的DEG数 Number of DEGs with GO function annotations	DEG总数 Total of DEG
清耕vs园艺地布Q_vs_F Clean tillage vs Horticultural ground cloth Q_vs_F	425	23	308	155	364
清耕vs生草栽培Q_vs_S Clean tillage vs Sod-culture Q_vs_S	119	15	132	57	108

图2 不同地面覆盖方式下新郁葡萄果皮KOG注释的分类统计注:(Note)A,RNA的加工与修饰(RNA processing and modification);C,能源的产生与转化(Energy production and conversion);D,细胞周期调控,细胞分裂,染色体分配(Cell cycle control,cell division,chromosome partitioning);E,(氨基酸转运与代谢)Amino acid transport and metabolism;F,(核苷酸转运与代谢)Nucleotide transport and metabolism;G,碳水化合物转运与代谢(Carbohydrate transport and metabolism);H,辅酶转运与代谢(Coenzyme transport and metabolism);I,脂质转运与代谢(Lipid transport and metabolism);J,翻译、核糖体结构和生物合成(Translation,ribosome structure and biosynthesis); K,转录(transcription);L,复制、重组和修复(Replication,recombination and repair);M,细胞壁/细胞膜的生物发生(Cell wall/membrane/envelope biogenesis);O,次生代谢物合成、转运和代谢(Posttranslational modification,protein turnover,chaperones);P,无机离子转运与代谢(Inorganic ion transport and metabolism);Q,次级代谢物的生物合成、转运和代谢(Secondary metabolites biosynthesis,transport and cetabolism);R,一般功能预测(General function prediction only);S,未知功能(Function Unknown);T,信号转导机制(Signal transduction mechanisms);U,胞质运输、分泌和囊泡运动(intracellular trafficking,secretion and vesicle transport);V,防御机制(Defense mechanisms); W,细胞外结构(xtracellular structures);Z, 细胞骨架(cytoskeleton)

Fig.2 KOG annotated histogram of classification statistics of grape peel by different ground mulch methods

图3 不同地面覆盖方式下葡萄果皮差异表达基因KEGG pathway代谢通路注(Note):横坐标表示注释到该通路的基因与有注释的基因总数的比例,纵坐标表示 KEGG 通路的名称。图形右侧的标签代表 KEGG 通路所属的分类(The abscissa represents the ratio of genes annotated to the pathway to the total number of annotated genes,and the ordinate represents the name of the KEGG pathway. The label on the right of the figure represents the classification to which the KEGG pathway belongs);环境信息处理(Environmental information processing);新陈代谢(Metabolism);有机系统(Organismal systems);MAPK信号通路-植物(MAPK signaling pathway-plant);植物激素信号转导(Plant hormone signal transduction);次级代谢物生物合成(Biosynthesis of secondary metabolites);类黄酮生物合成途径(Flavonoid biosynthesis);代谢途径(Metabolic pathways);苯丙烷类的代谢途径(Phenylalanine metabolism);苯丙烷类的生物合成(Phenylpropanoid biosynthesis);昼夜节律-植物(Circadian rhythm-plant);植物病原互作(Plant-pathogen interaction)。

Fig.3 Functional classification and pathway assignment of DEGs by KEGG by different mulch type

图4 不同地面覆盖方式下葡萄果皮差异表达基因富集散点图注(Note):纵坐标表示KEGG通路。横坐标表示Rich factor。Rich factor越大,富集的程度越大。点越大,通路富集的差异基因的数量越多。点的颜色越红,代表富集越显著(The ordinate represents the KEGG pathway. Abscissa represents the Rich Factor. The larger the Rich factor,the greater the degree of enrichment. Larger point indicates more number of differential genes enriched by pathway. Color shows significance with most significant in red);酪氨酸代谢(Tyrosine metabolism);莨菪烷、哌啶和吡啶生物碱的生物合成(Tropane,piperidine and pyridine alkaloid biosynthesis);硫胺素新陈代谢(Thiamine metabolism);牛磺酸和次牛磺酸代谢(Taurine and hypotaurine metabolism);二芳基庚烷和姜醇的生物合成(Stilbenoid,diarylheptanoid and gingerol biosynthesis);植物-病原互作(Plant-pathogen interaction);光合作用-天线蛋白(Photosynthesis-antenna proteins);光合作用(Photosynthesis);苯丙烷生物合成途径(Phenylpropanoid biosynthesis);苯丙烷类的代谢途径(Phenylalanine metabolism);代谢途径(Metabolic pathways);异黄酮生物合成途径(Isoquinoline alkaloid biosynthesis);甘油酯代谢(Glycerolipid metabolism);半乳糖代谢(Galactose metabolism);类黄酮生物合成途径(Flavonoid biosynthesis);二萜生物合成(Diterpenoid biosynthesis);昼夜节律-植物(Circadian rhythm-plant);次级代谢物生物合成(Biosynthesis of secondary metabolites);精氨酸和脯氨酸代谢(Arginine and proline metabolism);花青素生物合成(anthocyanin biosynthesis);各种类型的N -聚糖生物合成(Various types of N-glycan biosynthesis);色氨酸代谢(Tryptophan metabolism);酪氨酸代谢(Thiamine metabolism);类倍半萜烯和三萜(Sesquiterpenoid and triterpenoid);内质网中的蛋白质加工(Protein processing in endoplasmic reticulum);N-寡糖生物合成(N-Glycan biosynthesis);柠檬烯和蒎烯的降解(Limonene and pinene degradation);组氨酸代谢(Histidine metabolism);甘油酯代谢(Glycerolipid metabolism);脂肪酸延伸(Fatty acid elongation);角质、亚伯碱和蜡的生物合成(Cutin,suberine and wax biosynthesis);类胡萝卜素合成(Carotenoid biosynthesis);油菜素类固醇生物合成(Brassinosteroid biosynthesis);碱基切除修复(Base excision repair)。

Fig.4 Map of rich distribution points of differentially expressed genes of grape peel by different mulch types

表 4 不同地面覆盖方式下葡萄果皮黄酮代谢物组分和含量变化

Tab.4 Components and contents of flavonoid metabolites of grape peel by different mulch type(mg/kg)

代谢物ID Metabolite ID	物质 Compound	二级代谢产物 Class Ⅱ metabolite	清耕Q (对照,CK) Clean tillage		园艺地布 Horticultural ground cloth		生草栽培 Sod-culture
Flavonoid_145	原花青素B2	花青素	113.161±5.062^a (3.489%)		121.677±14.285^a (3.387%)		76.123±7.106^b (2.302%)
Flavonoid_65		查耳酮	17.546±1.872^a		18.213±2.625^a		15.875±0.745^a
Flavonoid_191	柚皮素查尔酮		0.198±0.058^b		0.345±0.048^a		0.186±0.049^b
Flavonoid_202			0.703±0.117^a		0.626±0.097^a		0.624±0.016^a
Flavonoid_22	新橙皮甙二氢查尔酮		0.023±0.017^ab		0.039±0.015^a		0.004±0.002^b
Flavonoid_31	柚皮苷二氢查尔酮		0.010±0.004^a		0.011±0.002^a		0.006±0.002^a
Flavonoid_177	总含量		0.008±0.001^b 18.848±1.946^a (0.57%)		0.018±0.004^a 19.253±2.706^a (0.54%)		0.005±0.001^b 16.700±0.683^a (0.51%)
Flavonoid_195	没食子儿茶素(-)	黄烷醇类	718.783±77.192^b		792.625±48.743^ab		903.325±68.297^a
Flavonoid_147	(-)-表儿茶素		397.187±41.006^a		455.475±48.453^a		383.830±17.308^a
Flavonoid_55	(-)-儿茶素		385.300±6.333^a		328.490±16.378^b		404.035±10.757^a
Flavonoid_84	(-)-Epigallocatechin		248.105±37.000^c		303.825±4.475^b		393.096±23.182^a
Flavonoid_150	儿茶素没食子酸酯(-)		9.069±1.240^b		10.132±0.997^b		12.403±0.370^a
Flavonoid_80	没食子儿茶素没食子酸酯(-) 总含量		6.282±1.771^b 1 764.726±137.344^b (54.41%)		7.469±0.628^b 1 898.014±115.200^ab (52.83%)		10.787±1.140^a 2 107.475±103.044^a (63.73%)
Flavonoid_176	柚皮素7-O-葡萄糖苷	二氢黄酮	2.280±0.397^a		2.186±0.387^a		1.484±0.028^b
Flavonoid_126	Eriodictyol		0.184±0.026^b		0.263±0.046^a		0.140±0.020^b
Flavonoid_78	圣草次甙		0.043±0.005^ab		0.047±0.014^a		0.026±0.006^b
Flavonoid_108	Pinocembrin		0.026±0.022^a 2.533±0.715^a (0.08%)		0.008±0.001^a 2.503±0.631^a (0.07%)		0.016±0.009^b 1.665±0.024^b (0.05%)
Flavonoid_01	落新妇苷	二氢黄酮醇	232.382±48.012^ab		262.966±16.701^a		176.495±16.215^b
Flavonoid_117	黄杞苷		24.027±6.243^b		33.018±2.330^a		16.988±1.595^b
Flavonoid_56	花旗松素		2.236±0.037^a		2.327±0.048^a		1.735±0.226^b
Flavonoid_40	二氢杨梅素		1.369±0.254^a		1.607±0.221^a		0.668±0.070^b
Flavonoid_139	二氢山奈酚		0.240±0.034^a		0.251±0.055^a		0.208±0.019^a
Flavonoid_04	异水飞蓟宾总含量		0.008±0.001^b 260.262±54.785^ab (8.03%)		0.011±0.002^a 300.181±18.845^a (8.36%)		0.000±0.000^c 196.094±17.889^b (5.93%)
Flavonoid_161	地奥司明	黄酮	82.793±9.443^b		135.293±20.719^a		43.813±5.059^c
Flavonoid_42	Nicotiflorin		13.405±2.472^b		20.647±2.846^a		6.015±0.631^c
Flavonoid_118	水仙苷		2.515±0.464^bA		4.043±0.852^a		1.741±0.480^b
Flavonoid_90	木犀草苷		2.121±0.035^a		1.932±0.411^a		1.790±0.241^a
Flavonoid_203	柠檬黄素		0.928±0.105^b		1.658±0.296^a		0.576±0.179^b
Flavonoid_165	高车前苷		0.217±0.017^a		0.201±0.026^a		0.157±0.014^b
Flavonoid_160	芹菜素7-葡萄糖苷		0.048±0.006^a		0.039±0.016^ab		0.014±0.014^b
Flavonoid_187	Tricetin		0.025±0.007^a		0.032±0.006^a		0.037±0.012^a
Flavonoid_50	木犀草素		0.007±0.002^a		0.008±0.001^a		0.009±0.004^a
Flavonoid_61	Chrysin		0.007±0.013^a		0.000±0.000^a		0.001±0.001^a
Flavonoid_06	香叶木素总含量		0.002±0.001^a 102.069±12.266^b (3.15%)		0.003±0.001^a 163.857±24.220^a (4.56%)		0.002±0.001^a 54.156±5.802^c (1.64%)
Flavonoid_02	槲皮素-3-O-葡萄糖醛酸苷	黄酮醇	705.296±59.981^ab		726.203±15.629^a		645.872±30.336^b
Flavonoid_98	金丝桃苷		84.123±16.962^ab		100.957±22.073^a		52.459±10.554^b
Flavonoid_57	芦丁		72.699±16.037^ab		86.735±11.412^a		51.383±3.789^b
Flavonoid_115	槲皮苷		42.880±9.875^a		39.482±4.102^ab		29.499±1.936^b
Flavonoid_197	异鼠李素-3-O-葡萄糖苷		35.671±4.306^b		65.029±19.704^a		19.015±4.202^b
Flavonoid_52	堪非醇3-新橙皮糖苷		5.296±1.359^a		5.693±0.679^a		2.181±0.177^b
Flavonoid_119	紫云英苷		4.158±0.833^ab		6.389±2.894^a		1.958±1.081^b
Flavonoid_194	槲皮素-7-O-β-D-葡萄糖苷		1.165±0.196^ab		1.420±0.329^a		0.878±0.168^b
Flavonoid_138	Baimaside		0.840±0.196^b		1.627±0.336^a		0.317±0.068^c
Flavonoid_54	异鼠李素-3-O-新橙皮苷		0.486±0.020^a		0.618±0.124^a		0.205±0.040^b
Flavonoid_23	槲皮素		0.326±0.043^b		0.537±0.090^a		0.285±0.018^b
Flavonoid_178	杨梅素		0.193±0.025^b		0.516±0.169^a		0.124±0.064^b
Flavonoid_92	杨梅素			0.167±0.044^b		0.518±0.199^a		0.063±0.018^b
Flavonoid_175	Afzelin		0.117±0.050^a		0.134±0.035^a		0.037±0.023^b
Flavonoid_58	异鼠李素		0.092±0.010^b		0.207±0.041^a		0.066±0.003^b
Flavonoid_188	西伯利亚落叶松黄酮		0.000±0.000^b		0.058±0.007^a		0.000±0.000^b
Flavonoid_123	2'-O-没食子酰基金丝桃苷总含量		0.000±0.000^a 953.342±39.099 ^ab (29.40%)		0.14±0.024^a 1036.137±62.291^a (28.84%)		0.000±0.000^a 804.342±39.099^b (24.32%)
Flavonoid_153	Glycitin	异黄酮	0.614±0.143^a		0.691±0.070^a		0.726±0.065^a
Flavonoid_73	Genistin		0.220±0.039^a		0.246±0.095^a		0.147±0.034^a
Flavonoid_96	毛蕊异黄酮苷		0.005±0.005^a 0.839±0.185^a (0.03%)		0.000±0.000^a 0.937±0.028^a (0.03%)		0.000±0.000^a 0.873±0.035^a (0.03%)
Flavonoid_122	7-Hydroxy-4H-chromen-4-one	酚酸类	0.396±0.116^a		0.302±0.085^a		0.395±0.050^a
Flavonoid_79	女贞苷		0.052±0.012^a		0.045±0.010^a		0.031±0.013^a
Flavonoid_35	丁香醛总含量		0.000±0.000^a 0.448±0.124^a (0.01%)		0.000±0.000^a 0.347±0.077^a (0.01%)		0.153±0.266^a 0.580±0.236^a (0.02%)
Flavonoid_157	异芒果苷	口山酮	0.024±0.004^a (0.00%)		0.021±0.001^a (0.00%)		0.021±0.001^a (0.00%)
Flavonoid_102	茶黄素	-	27.176±3.262^b (0.84%)		49.820±1.176^a (1.39%)		48.822±5.892^a (1.48%)
Flavonoid_154	黄酮碳糖苷	-	0.007±0.002^a (0.00%)		0.003±0.001^b (0.00%)		0.009±0.001^a (0.00%)

表 4 不同地面覆盖方式下葡萄果皮黄酮代谢物组分和含量变化

Tab.4 Components and contents of flavonoid metabolites of grape peel by different mulch type(mg/kg)

代谢物ID Metabolite ID	物质 Compound	二级代谢产物 Class Ⅱ metabolite	清耕Q (对照,CK) Clean tillage		园艺地布 Horticultural ground cloth		生草栽培 Sod-culture
Flavonoid_145	原花青素B2	花青素	113.161±5.062^a (3.489%)		121.677±14.285^a (3.387%)		76.123±7.106^b (2.302%)
Flavonoid_65		查耳酮	17.546±1.872^a		18.213±2.625^a		15.875±0.745^a
Flavonoid_191	柚皮素查尔酮		0.198±0.058^b		0.345±0.048^a		0.186±0.049^b
Flavonoid_202			0.703±0.117^a		0.626±0.097^a		0.624±0.016^a
Flavonoid_22	新橙皮甙二氢查尔酮		0.023±0.017^ab		0.039±0.015^a		0.004±0.002^b
Flavonoid_31	柚皮苷二氢查尔酮		0.010±0.004^a		0.011±0.002^a		0.006±0.002^a
Flavonoid_177	总含量		0.008±0.001^b 18.848±1.946^a (0.57%)		0.018±0.004^a 19.253±2.706^a (0.54%)		0.005±0.001^b 16.700±0.683^a (0.51%)
Flavonoid_195	没食子儿茶素(-)	黄烷醇类	718.783±77.192^b		792.625±48.743^ab		903.325±68.297^a
Flavonoid_147	(-)-表儿茶素		397.187±41.006^a		455.475±48.453^a		383.830±17.308^a
Flavonoid_55	(-)-儿茶素		385.300±6.333^a		328.490±16.378^b		404.035±10.757^a
Flavonoid_84	(-)-Epigallocatechin		248.105±37.000^c		303.825±4.475^b		393.096±23.182^a
Flavonoid_150	儿茶素没食子酸酯(-)		9.069±1.240^b		10.132±0.997^b		12.403±0.370^a
Flavonoid_80	没食子儿茶素没食子酸酯(-) 总含量		6.282±1.771^b 1 764.726±137.344^b (54.41%)		7.469±0.628^b 1 898.014±115.200^ab (52.83%)		10.787±1.140^a 2 107.475±103.044^a (63.73%)
Flavonoid_176	柚皮素7-O-葡萄糖苷	二氢黄酮	2.280±0.397^a		2.186±0.387^a		1.484±0.028^b
Flavonoid_126	Eriodictyol		0.184±0.026^b		0.263±0.046^a		0.140±0.020^b
Flavonoid_78	圣草次甙		0.043±0.005^ab		0.047±0.014^a		0.026±0.006^b
Flavonoid_108	Pinocembrin		0.026±0.022^a 2.533±0.715^a (0.08%)		0.008±0.001^a 2.503±0.631^a (0.07%)		0.016±0.009^b 1.665±0.024^b (0.05%)
Flavonoid_01	落新妇苷	二氢黄酮醇	232.382±48.012^ab		262.966±16.701^a		176.495±16.215^b
Flavonoid_117	黄杞苷		24.027±6.243^b		33.018±2.330^a		16.988±1.595^b
Flavonoid_56	花旗松素		2.236±0.037^a		2.327±0.048^a		1.735±0.226^b
Flavonoid_40	二氢杨梅素		1.369±0.254^a		1.607±0.221^a		0.668±0.070^b
Flavonoid_139	二氢山奈酚		0.240±0.034^a		0.251±0.055^a		0.208±0.019^a
Flavonoid_04	异水飞蓟宾总含量		0.008±0.001^b 260.262±54.785^ab (8.03%)		0.011±0.002^a 300.181±18.845^a (8.36%)		0.000±0.000^c 196.094±17.889^b (5.93%)
Flavonoid_161	地奥司明	黄酮	82.793±9.443^b		135.293±20.719^a		43.813±5.059^c
Flavonoid_42	Nicotiflorin		13.405±2.472^b		20.647±2.846^a		6.015±0.631^c
Flavonoid_118	水仙苷		2.515±0.464^bA		4.043±0.852^a		1.741±0.480^b
Flavonoid_90	木犀草苷		2.121±0.035^a		1.932±0.411^a		1.790±0.241^a
Flavonoid_203	柠檬黄素		0.928±0.105^b		1.658±0.296^a		0.576±0.179^b
Flavonoid_165	高车前苷		0.217±0.017^a		0.201±0.026^a		0.157±0.014^b
Flavonoid_160	芹菜素7-葡萄糖苷		0.048±0.006^a		0.039±0.016^ab		0.014±0.014^b
Flavonoid_187	Tricetin		0.025±0.007^a		0.032±0.006^a		0.037±0.012^a
Flavonoid_50	木犀草素		0.007±0.002^a		0.008±0.001^a		0.009±0.004^a
Flavonoid_61	Chrysin		0.007±0.013^a		0.000±0.000^a		0.001±0.001^a
Flavonoid_06	香叶木素总含量		0.002±0.001^a 102.069±12.266^b (3.15%)		0.003±0.001^a 163.857±24.220^a (4.56%)		0.002±0.001^a 54.156±5.802^c (1.64%)
Flavonoid_02	槲皮素-3-O-葡萄糖醛酸苷	黄酮醇	705.296±59.981^ab		726.203±15.629^a		645.872±30.336^b
Flavonoid_98	金丝桃苷		84.123±16.962^ab		100.957±22.073^a		52.459±10.554^b
Flavonoid_57	芦丁		72.699±16.037^ab		86.735±11.412^a		51.383±3.789^b
Flavonoid_115	槲皮苷		42.880±9.875^a		39.482±4.102^ab		29.499±1.936^b
Flavonoid_197	异鼠李素-3-O-葡萄糖苷		35.671±4.306^b		65.029±19.704^a		19.015±4.202^b
Flavonoid_52	堪非醇3-新橙皮糖苷		5.296±1.359^a		5.693±0.679^a		2.181±0.177^b
Flavonoid_119	紫云英苷		4.158±0.833^ab		6.389±2.894^a		1.958±1.081^b
Flavonoid_194	槲皮素-7-O-β-D-葡萄糖苷		1.165±0.196^ab		1.420±0.329^a		0.878±0.168^b
Flavonoid_138	Baimaside		0.840±0.196^b		1.627±0.336^a		0.317±0.068^c
Flavonoid_54	异鼠李素-3-O-新橙皮苷		0.486±0.020^a		0.618±0.124^a		0.205±0.040^b
Flavonoid_23	槲皮素		0.326±0.043^b		0.537±0.090^a		0.285±0.018^b
Flavonoid_178	杨梅素		0.193±0.025^b		0.516±0.169^a		0.124±0.064^b
Flavonoid_92	杨梅素			0.167±0.044^b		0.518±0.199^a		0.063±0.018^b
Flavonoid_175	Afzelin		0.117±0.050^a		0.134±0.035^a		0.037±0.023^b
Flavonoid_58	异鼠李素		0.092±0.010^b		0.207±0.041^a		0.066±0.003^b
Flavonoid_188	西伯利亚落叶松黄酮		0.000±0.000^b		0.058±0.007^a		0.000±0.000^b
Flavonoid_123	2'-O-没食子酰基金丝桃苷总含量		0.000±0.000^a 953.342±39.099 ^ab (29.40%)		0.14±0.024^a 1036.137±62.291^a (28.84%)		0.000±0.000^a 804.342±39.099^b (24.32%)
Flavonoid_153	Glycitin	异黄酮	0.614±0.143^a		0.691±0.070^a		0.726±0.065^a
Flavonoid_73	Genistin		0.220±0.039^a		0.246±0.095^a		0.147±0.034^a
Flavonoid_96	毛蕊异黄酮苷		0.005±0.005^a 0.839±0.185^a (0.03%)		0.000±0.000^a 0.937±0.028^a (0.03%)		0.000±0.000^a 0.873±0.035^a (0.03%)
Flavonoid_122	7-Hydroxy-4H-chromen-4-one	酚酸类	0.396±0.116^a		0.302±0.085^a		0.395±0.050^a
Flavonoid_79	女贞苷		0.052±0.012^a		0.045±0.010^a		0.031±0.013^a
Flavonoid_35	丁香醛总含量		0.000±0.000^a 0.448±0.124^a (0.01%)		0.000±0.000^a 0.347±0.077^a (0.01%)		0.153±0.266^a 0.580±0.236^a (0.02%)
Flavonoid_157	异芒果苷	口山酮	0.024±0.004^a (0.00%)		0.021±0.001^a (0.00%)		0.021±0.001^a (0.00%)
Flavonoid_102	茶黄素	-	27.176±3.262^b (0.84%)		49.820±1.176^a (1.39%)		48.822±5.892^a (1.48%)
Flavonoid_154	黄酮碳糖苷	-	0.007±0.002^a (0.00%)		0.003±0.001^b (0.00%)		0.009±0.001^a (0.00%)

图5 不同地面覆盖方式下葡萄果皮差异代谢物KEGG富集图注(Note):纵坐标表示每个通路对应的Rich factor,纵坐标为通路名称,点的颜色为P value,越红表示富集越显著。点的大小代表富集到的差异代谢物的个数多少(The ordinate represents the Rich factor corresponding to each pathway,the ordinate represents the pathway name,and the color of the point is P value. The redder indicates the more significant enrichment. The size of the dot represents the number of enriched differential metabolites);类黄酮生物合成(Flavonoid biosynthesis); 黄酮和黄烷醇代谢物生物合成(Flavone and flavonol biosynthesis); 次级代谢物生物合成(Biosynthesis of secondary metabolites)

Fig.5 KEGG enrichment map of differential metabolites of grape peel by different mulch type

图6 不同地面覆盖方式下葡萄果皮差异代谢物差异倍数

Fig.6 The difference multiples analysis of anthocyanin by different mulch type

表5 新郁葡萄果皮黄酮生物合成途径中候选基因

Tab.5 Candidate gene involved in flavonoid biosynthesis pathway in Xinyu grape peel

序号 No.	清耕vs园艺地布Q_vs_F Clean tillage vs Horticultural ground cloth Q_vs_F
序号 No.	生物合成途径 Biosynthesis	名称 Name	缩写 Abbreviations	KEGG注释到 unigene数量 KEGG notes the number of unigene	备注 Note
1	类黄酮代谢生物合成	反式肉桂酸-4-单加氧酶	CYP73A(C4H)	1	上调
2		查尔酮合成酶	CHS	6	上调
3		黄酮-3-脱氢酶	F3H	6	上调
4		无色花色素还原酶	LAR	3	上调
5		莽草酸羟基肉桂酰基转移酶	HCT	4	上调和下调
6		黄酮醇合成酶	FLS	4	上调和下调
7		黄烷酮-7-氧-葡萄糖苷 2″-氧-β-L-鼠李糖基转移酶	C12RT1	2	下调
8	黄酮和黄酮醇生物合成	黄烷酮-7-氧-葡萄糖苷 2″-氧-β-L-鼠李糖基转移酶	C12RT1	2	下调
序号 No.	清耕vs生草栽培Q_vs_S Clean tillage vs Sod-culture Q_vs_S
1	类黄酮代谢生物合成	查尔酮合成酶	CHS	6	下调
2		黄酮-3-脱氢酶	F3H	6	下调
3		类黄酮3',5'异构酶	CYP75A(F3',5'H)	8	下调
4		黄酮醇合成酶	FLS	3	下调
5		无色花色素还原酶	LAR	3	下调
6	黄酮和黄酮醇生物合成	类黄酮3',5'异构酶	CYP75A(F3',5'H)	3	下调

图6 不同地面覆盖方式下葡萄果皮差异代谢物差异倍数

Fig.6 The difference multiples analysis of anthocyanin by different mulch type

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