基于非靶向代谢组学的上部烟叶不同成熟度差异分析

doi:10.13304/j.nykjdb.2024.0226

摘要/Abstract

摘要：

为探究不同采收成熟度对上部叶的内在质量和代谢组学的影响，以云烟87为试验材料，采收叶片SPAD值分别为30~35（BM1）、25~30（BM2）、20~25（BM3）、15~20（BM4）的烤烟上部叶，研究不同采收成熟度对上部叶农艺性状、化学成分协调性、感官质量以及代谢组学的影响。结果显示，BM3处理上部叶的化学成分协调性较好，感官评吸分数最高。4种处理共筛选出230种差异代谢物，主要是有机酸类、生物碱类、糖类、氨基酸类、醇类和核苷类等。生物信息学分析表明，差异代谢物中的L-色氨酸、绿原酸、棕榈油酸、刀豆氨酸、肌酸、山梨醇、葫芦巴碱、α-亚麻酸、甘油酸、创伤性酸、胆碱、马泰树脂醇、脱氧肌苷、芸香苷、L-苯丙氨酸、鸟嘌呤、L-脯氨酸、L-蛋氨酸、葡糖二酸、尼古丁、油酸、L-丝氨酸、黄氧酸、L-谷氨酸、丁香酸、丁香酚、色胺酸等可能是上部烟叶成熟过程中较关键的代谢物。差异代谢物显著富集在氨基酸代谢、核苷酸代谢、碳水化合物代谢、维生素代谢和脂质代谢等通路中，其中甘氨酸、丝氨酸和苏氨酸的代谢，苯丙烷类生物合成，丙氨酸、天冬氨酸和谷氨酸代谢等通路在烟叶成熟过程中油分和香气物质的形成起重要调控作用。

关键词: 烟叶, 代谢组学, 成熟度, 差异代谢物

Abstract:

To explore the effects of different harvest maturity on internal quality of upper leaves and metabolomics of tobacco， Yunyan 87 was used as the experimental material， and the leaves were sampled with SPAD value of 30~35 （BM1）， 25~30 （BM2）， 20~25 （BM3）， 15~20 （BM4）. The effects of different harvest maturity on the agronomic traits， chemical composition coordination， sensory quality and metabolomics of upper leaves were analyzed. The results showed that the upper leaves of BM3 treatment had better coordination of chemical components and the highest sensory evaluation score. A total of 230 differential metabolites were screened from 4 treatments， mainly including organic acids， alkaloids， sugars， amino acids， alcohols and nucleosides. Bioinformatics analysis showed that L-tryptophan， chlorogenic acid， palmitoleic acid， concanavalin， creatine， sorbitol， trigonelline， α-linolenic acid， glyceric acid， traumatic acid， choline， matairesinol， deoxyinosine， rutin， L-phenylalanine， guanine， L-proline， L-methionine， glucaric acid， nicotine， oleic acid， L-serine， flavic acid， L-glutamic acid， syringic acid， eugenol and tryptophan might be the key metabolites during the maturation of upper tobacco leaves. Differential metabolites were significantly enriched in pathways such as amino acid metabolism， nucleotide metabolism， carbohydrate metabolism， vitamin metabolism and lipid metabolism. Among them， glycine， serine and threonine metabolism， phenylpropanoid biosynthesis， alanine， aspartic acid and glutamic acid metabolism pathways played an important regulatory role in the formation of oil and aroma substances during tobacco leaf maturation.

Key words: tobacco leaf, metabolomics, maturity, differential metabolites

中图分类号:

S572

张彩虹, 张力, 王卫民, 赵炯平, 韩丹, 许自成, 张仲文, 邵惠芳. 基于非靶向代谢组学的上部烟叶不同成熟度差异分析[J]. 中国农业科技导报, 2024, 26(10): 58-70.

Caihong ZHANG, Li ZHANG, Weimin WANG, Jiongping ZHAO, Dan HAN, Zicheng XU, Zhongwen ZHANG, Huifang SHAO. Difference Analysis of Different Maturity of Upper Tobacco Leaves Based on Non-targeted Metabolomics[J]. Journal of Agricultural Science and Technology, 2024, 26(10): 58-70.

图/表 12

参考文献 36

1	张树堂,杨雪彪,王亚辉,等.不同成熟度烤烟鲜烟叶的组织结构比较[J].烟草科技,2005,38(1):38-40.
	ZHANG S T, YANG X B, WANG Y H,et al.. Comparison of tissue of fresh flue-cured tobacco leaves with different maturity [J].Tob. Sci. Technol., 2005,38(1):38-40.
2	贾中林,尹启生,戴华鑫,等.烤烟上部烟叶成熟生理特性研究进展[J].烟草科技,2022,55(7):99-112.
	JIA Z L, YIN Q S, DAI H X,et al.. Research progress on physiological characteristics of mature upper leaves of flue-cured tobacco [J]. Tob. Sci. Technol.,2022,55(7):99-112.
3	孙立娟,李虎林,金哲,等.不同成熟度烤烟外观特征及化学成分的变化[J].湖北农业科学,2008,47(3):318-320.
	SUN L J, LI H L, JIN Z,et al.. Study on the main chemical component of flue-cured tobacco during maturity stage [J].Hubei Agric. Sci., 2008,47(3):318-320.
4	刘海业.采收时期对烤烟成熟特性及产质量的影响[D].广州:华南农业大学,2016.
	LIU H Y. Effects of harvest time on maturity characteristics, yield and quality of flue-cured tobacco [D]. Guangzhou: South China Agricultural University, 2016.
5	王勇,周冀衡,肖志新,等.不同成熟度对烤烟烟叶品质和安全性指标的影响[J].中国烟草科学,2007,28(3):26-29.
	WANG Y, ZHOU J H, XIAO Z X, et al.. Influence of the different maturity on indexes of quality and security for flue-cured tobacco leaves [J]. Chin. Tob. Sci., 2007,28(3):26-29.
6	李迪秦,龚湛武,陈一凡,等.不同播种移栽期对烤烟产量与品质影响的多指标模糊评价[J].核农学报,2017,31(11):2258-2264.
	LI D Q, GONG Z W, CHEN Y F,et al.. Multi-index fuzzy evaluation of the effect of different sowing and transplanting date on flue-cured tobacco yield and quality [J]. J. Nucl. Agric.Sci., 2017,31(11):2258-2264.
7	周健民,沈仁芳.土壤学大辞典[M].北京:科学出版社,2013:1-902.
	ZHOU J M, SHEN R F. Dictionary of Soil Science [M].Beijing:Science Press,2013:1-902.
8	ZI X, ZHOU S, WU B. Alpha-linolenic acid mediates diverse drought responses in maize (Zea mays L.) at seedling and flowering stages [J/OL].Molecules,2022,27(3):771 [2024-02-20]. .
9	袁俊,盛莎莎,刘荣鹏,等.镉胁迫对丹参生理特性和代谢特征的影响[J].植物科学学报,2022,40(3):408-417.
	YUAN J, SHENG S S, LIU R P, et al.. Effects of cadmium on physiological characteristics and metabolic profiles of Salvia miltiorrhiza Bunge [J]. Plant Sci. J., 2022,40(3):408-417.
10	彭翠云,汪海燕,史宏志,等.延迟采收对豫中烤烟上部叶生理指标和代谢组学的影响[J].核农学报,2021,35(11):2655-2663.
	PENG C Y, WANG H Y, SHI H Z, et al.. Physiological indexes and metabolomics changes in the upper stalk tobacco leaves under the delaying harvesting in Yuzhong area [J]. J. Nucl. Agric. Sci., 2021,35(11):2655-2663.
11	高云,许自成.江西烤烟生育期主要气象灾害及防御措施分析[J].山西农业科学,2017,45(10):1686-1690.
	GAO Y, XU Z C. Analysis of main meteorological disasters and prevention measures during the growth period of flue cured tobacco in Jiangxi [J]. J.Shanxi Agric.Sci.,2017,45(10):1686-1690.
12	孙阳阳,靳志伟,黄明迪,等.SPAD值与鲜烟叶成熟度及烤后烟叶质量的关系[J].中国烟草科学,2016,37(2):42-46.
	SUN Y Y, JIN Z W, HUANG M D, et al.. The relationship of SPAD value,maturity of fresh tobacco leaves and quality of flue-cured tobacco leaves [J]. Chin. Tob. Sci.,2016,37(2):42-46.
13	陈乾锦,池国胜,袁帅,等.鲜烟不同采收成熟度和SPAD值与烤后烟叶质量关系研究[J].江西农业学报,2021,33(1):55-59.
	CHEN Q J, CHI G S, YUAN S, et al.. Study on relationship among harvest maturity, SPAD values and quality of flue-cured tobacco after curing [J]. Acta Agric. Jiangxi, 2021,33(1):55-59.
14	张威,刘楠,马雁军,等. 烟草及烟草制品总植物碱的测定连续流动(硫氰酸钾)法: [S].北京:中国标准出版社,2022.
15	刘惠民,王芳,李荣,等. 烟草及烟草制品总氮的测定连续流动法: [S].北京:中国标准出版社,2004.
16	何声宝,张威,罗安娜,等. 烟草及烟草制品水溶性糖的测定连续流动法: [S].北京:中国标准出版社,2019.
17	张威,王芳,王颖,等. 烟草及烟草制品钾的测定连续流动法: [S].北京:中国标准出版社,2007.
18	国家烟草专卖局. 烟草及烟草制品氯的测定连续流动法: [S].北京:中国标准出版社,2011.
19	罗登山,王兵,马宇平,等. 烤烟烟叶质量风格特色感官评价方法: [S].北京:中国标准出版社,2015.
20	VASILEV N, BOCCARD J, LANG G,et al..Structured plant metabolomics for the simultaneous exploration of multiple factors [J/OL]. Sci. Rep., 2016, 6:37390 [2016-11-17]. .
21	PRABHU A H, KANT S, KESARWANI P,et al..Integrative cross-platform analyses identify enhanced heterotrophy as a metabolic hallmark in glioblastoma [J]. Neuro-Oncology,2019,21(3):337-347.
22	杨洋,符云鹏,刘新源,等.豫西烟区上部叶成熟度与品质形成的关系[J].江苏农业科学,2023,51(8):79-85.
23	刘国顺.烟草栽培学[M].北京:中国农业出版社,2003:1-335.
	LIU G S.Tobacco Cultivation [M].Beijing:China Agriculture Press,2003:1-335.
24	樊希彬,李丹,左晓晴,等.气候条件对烟草生长的影响分析[J].黑龙江农业科学,2016(4):27-30.
	FAN X B, LI D, ZUO X Q, et al.. Effect of climate conditions on tobacco growth [J]. Heilongjiang Agric. Sci., 2016(4):27-30.
25	宋笑龙,刘芳,宗胜杰,等.不同采收成熟度对烤烟萜类致香化合物及烟叶品质的影响[J].江苏农业科学,2024,52(2):104-111.
26	关辉,周艳宾,朱宏强,等.采收成熟度对烤烟中上部烟叶品质的影响[J].南方农业,2021,15(24):1-3, 7.
27	张丽,姬厚伟,黄锡娟,等.植物代谢组学及其在烟草上的应用进展[J].中国烟草学报,2015,21(5):126-134.
	ZHANG L, JI H W, HUANG X J, et al.. Plant metabolomics and its application in tobacco [J].Acta Tab. Sin., 2015,21(5):126-134.
28	向海英,刘欣,逄涛,等.基于非靶向代谢组学的红花大金元不同生育期烟叶代谢图谱差异分析[J].中国烟草学报,2022,28(4):77-84.
	XIANG H Y, LIU X, PANG T, et al.. Differential analysis of metabolic profiles of Honghua Dajinyuan tobacco leaves in different growth stage based on untargeted metabolomics [J].Acta Tab. Sin.,2022,28(4):77-84.
29	BAN Q, WANG X, PAN C, et al.. Comparative analysis of the response and gene regulation in cold resistant and susceptible tea plants [J/OL].PLoS One,2017,12(12):e0188514 [2017-12-16]. .
30	梁太波,张景玲,田雷,等.干旱胁迫下外源甜菜碱和脯氨酸对烤烟抗氧化代谢的影响[J].烟草科技,2013,46(2):68-71.
	LIANG T B, ZHANG J L, TIAN L, et al.. Effects of exogenous glycine betaine and proline on antioxidant metabolism of flue-cured tobacco under drought stress [J]. Tob. Sci. Technol.,2013,46(2):68-71.
31	宋永骏,刁倩楠,齐红岩.多胺代谢与植物抗逆性研究进展[J].中国蔬菜,2012(18):36-42.
	SONG Y J, DIAO Q N, QI H Y. Research progress on relationship between polyamine metabolism and plant stress resistance [J]. China Veget., 2012(18):36-42.
32	武模戈,袁刘正.多胺与植物抗逆性关系研究进展[J].安徽农业科学,2008,36(9):3516-3518.
	WU M G, YUAN L Z. Research progress in the relationship between polyamine and plant resistance [J]. J. Anhui Agric. Sci., 2008,36(9):3516-3518.
33	OLIVA M, GUY A, GALILI G,et al.. Enhanced production of aromatic amino acids in tobacco plants leads to increased phenylpropanoid metabolites and tolerance to stresses [J/OL].Plant Sci.,2020,11:604349 [2021-1-12]. .
34	王威威,席飞虎,杨少峰,等.烟草烟碱合成代谢调控研究进展[J].亚热带农业研究,2016,12(1):62-67.
	WANG W W, XI F H, YANG S F, et al.. Progress on nicotine metabolism regulation in tobacco [J]. Subtrop.Agric.Res.,2016,12(1):62-67.
35	ZHANG L, ZHANG X T, JI H W, et al.. Metabolic profiling of tobacco leaves at different growth stages or different stalk positions by gas chromatography-mass spectrometry [J].Ind.Crops Prod., 2018,116:46-55.
36	贾中林,郑庆霞,戴华鑫,等.烤烟上部叶成熟过程中代谢组的差异分析[J].烟草科技,2023,56(1):1-10.
	JIA Z L, ZHENG Q X, DAI H X, et al.. Differential analysis of metabolome of upper flue-cured tobacco leaf during ripening process [J]. Tob. Sci. Technol., 2023,56(1):1-10.

指标Index		日期Date （m/d）
指标Index		6/4	6/10	6/16	6/23
长势Growth	株高Plant height/cm	99.69±4.82 a	100.40±5.68 a	100.60±5.56 a	101.16±5.14 a
	茎围Stem circumference/cm	9.16±0.47 a	9.17±0.76 a	9.17±0.76 a	9.20±0.12 a
	有效叶片数Effective leaf number	13.80±0.84 a	13.00±1.04 ab	10.00±1.7 ab	8.00±4.04 b
顶1 叶Top 1 leaf	叶长Leaf length/cm	55.64±3.54 a	56.23±2.13 a	58.93±0.90 a	59.53±4.10 a
	叶宽Leaf width/cm	17.36±3.31 a	17.86±2.34 a	20.00±2.29 a	20.23±3.46 a
	叶面积Leaf area/cm²	965.91±119.80 a	1 004.26±123.75 a	1 178.60±121.91 a	1 204.29±362.19 a
顶3叶 Top 3 leaf	叶长Leaf length/cm	65.31±5.83 a	65.73±4.84 a	66.50±4.82 a	66.56±6.62 a
	叶宽Leaf width/cm	20.23±1.98 b	20.86±1.03 ab	22.33±0.28 ab	23.21±1.17 a
	叶面积Leaf area/cm²	1 321.22±114.80 a	1 371.12±145.79 a	1 484.94±97.06 a	1 544.85±215.26 a
顶6叶 Top 6 leaf	叶长Leaf length/cm	71.86±4.56 a	72.50±4.09 a	73.86±2.96 a	72.96±4.33 a
	叶宽Leaf width/cm	28.10±1.75 b	30.33±0.57 a	30.83±0.28 a	30.86±1.02 a
	叶面积Leaf area/cm²	2 019.26±170.30 a	2 198.92±158.05 a	2 277.10±79.56 a	2 251.54±198.64 a

指标Index		日期Date （m/d）
指标Index		6/4	6/10	6/16	6/23
长势Growth	株高Plant height/cm	99.69±4.82 a	100.40±5.68 a	100.60±5.56 a	101.16±5.14 a
	茎围Stem circumference/cm	9.16±0.47 a	9.17±0.76 a	9.17±0.76 a	9.20±0.12 a
	有效叶片数Effective leaf number	13.80±0.84 a	13.00±1.04 ab	10.00±1.7 ab	8.00±4.04 b
顶1 叶Top 1 leaf	叶长Leaf length/cm	55.64±3.54 a	56.23±2.13 a	58.93±0.90 a	59.53±4.10 a
	叶宽Leaf width/cm	17.36±3.31 a	17.86±2.34 a	20.00±2.29 a	20.23±3.46 a
	叶面积Leaf area/cm²	965.91±119.80 a	1 004.26±123.75 a	1 178.60±121.91 a	1 204.29±362.19 a
顶3叶 Top 3 leaf	叶长Leaf length/cm	65.31±5.83 a	65.73±4.84 a	66.50±4.82 a	66.56±6.62 a
	叶宽Leaf width/cm	20.23±1.98 b	20.86±1.03 ab	22.33±0.28 ab	23.21±1.17 a
	叶面积Leaf area/cm²	1 321.22±114.80 a	1 371.12±145.79 a	1 484.94±97.06 a	1 544.85±215.26 a
顶6叶 Top 6 leaf	叶长Leaf length/cm	71.86±4.56 a	72.50±4.09 a	73.86±2.96 a	72.96±4.33 a
	叶宽Leaf width/cm	28.10±1.75 b	30.33±0.57 a	30.83±0.28 a	30.86±1.02 a
	叶面积Leaf area/cm²	2 019.26±170.30 a	2 198.92±158.05 a	2 277.10±79.56 a	2 251.54±198.64 a

处理Treatment	烟碱Nicotine/%	总氮Total nitrogen/%	总糖Total sugar/%	还原糖Reducing sugar/%	钾Potassium/%	氯 Chlorine/%	糖碱比 Sugar alkali ratio	钾氯比Potassium chloride ratio	两糖比Two sugar ratio
BM1	2.03±0.03 d	2.08±0.02 b	30.77±0.15 a	23.70±0.26 a	1.72±0.13 b	0.19±0.03 b	15.20±0.26 a	9.13±1.08 a	0.77±0.01 b
BM2	2.13±0.01 c	2.34±0.02 a	29.77±0.21 b	22.93±0.76 ab	1.75±0.16 b	0.24±0.02 b	13.98±0.15 b	7.19±0.46 b	0.77±0.02 b
BM3	2.28±0.05 b	1.79±0.03 c	29.10±0.30 c	21.93±0.42 bc	1.93±0.08 ab	0.40±0.05 a	12.77±0.19 c	4.84±0.37 c	0.75±0.01 b
BM4	2.37±0.02 a	1.82±0.02 c	26.63±0.21 d	21.37±0.55 c	2.02±0.08 a	0.37±0.04 a	11.22±0.16 d	5.45±0.43 c	0.80±0.02 a

处理Treatment	烟碱Nicotine/%	总氮Total nitrogen/%	总糖Total sugar/%	还原糖Reducing sugar/%	钾Potassium/%	氯 Chlorine/%	糖碱比 Sugar alkali ratio	钾氯比Potassium chloride ratio	两糖比Two sugar ratio
BM1	2.03±0.03 d	2.08±0.02 b	30.77±0.15 a	23.70±0.26 a	1.72±0.13 b	0.19±0.03 b	15.20±0.26 a	9.13±1.08 a	0.77±0.01 b
BM2	2.13±0.01 c	2.34±0.02 a	29.77±0.21 b	22.93±0.76 ab	1.75±0.16 b	0.24±0.02 b	13.98±0.15 b	7.19±0.46 b	0.77±0.02 b
BM3	2.28±0.05 b	1.79±0.03 c	29.10±0.30 c	21.93±0.42 bc	1.93±0.08 ab	0.40±0.05 a	12.77±0.19 c	4.84±0.37 c	0.75±0.01 b
BM4	2.37±0.02 a	1.82±0.02 c	26.63±0.21 d	21.37±0.55 c	2.02±0.08 a	0.37±0.04 a	11.22±0.16 d	5.45±0.43 c	0.80±0.02 a

处理Treatment	香气质Fragrance temperament	香气量Aroma amount	透发性Permeability	杂气Miscellaneous gas	细腻程度Delicate degree	柔和程度Degree of softness	圆润感Roundness feeling	刺激性Irritation	干燥感Dry feeling	余味Aftertaste	总分Total score
BM1	11.50	11.50	4.00	5.00	4.50	4.50	4.50	4.50	5.50	5.50	61.00
BM2	12.00	12.00	5.00	5.50	5.00	5.00	5.00	5.00	5.50	5.50	65.50
BM3	12.50	12.50	5.00	5.50	5.00	5.00	5.00	5.50	5.00	5.50	66.50
BM4	12.00	12.00	5.00	5.50	4.50	4.50	5.00	5.00	5.50	5.50	64.50