蚜害茶树挥发物组分变化的聚类分析

doi:10.11983/CBB21031

植物学报 ›› 2021, Vol. 56 ›› Issue (4): 422-432.DOI: 10.11983/CBB21031

蚜害茶树挥发物组分变化的聚类分析

孙廷哲^1†, 岂泽华^1†, 梁可欣¹, 李沁¹^,², 饶玉春²^,^*(), 穆丹¹^,^*()

¹安庆师范大学生命科学学院, 安庆 246133
²浙江师范大学化学与生命科学学院, 金华 321004

收稿日期:2021-02-07 接受日期:2021-04-19 出版日期:2021-07-01 发布日期:2021-06-30
通讯作者: 饶玉春,穆丹
作者简介:mudansmile@126.com
*E-mail: ryc@zjnu.cn;
第一联系人：
† 共同第一作者
基金资助:
国家自然科学基金(31971185);国家自然科学基金(31800316);安徽省高校优秀青年人才支持计划(gxyqZD2020031);安徽省高校优秀青年人才支持计划(gxyq2018034);安徽省教育厅自然基金(KJ2017A359)

Clustering Analysis of Volatile Components from the Tea Plants Infested by Tea Aphid (Toxoptera aurantii)

Tingzhe Sun^1†, Zehua Qi^1†, Kexin Liang¹, Qin Li¹^,², Yuchun Rao²^,^*(), Dan Mu¹^,^*()

¹School of Life Sciences, Anqing Normal University, Anqing 246133, China
²School of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China

Received:2021-02-07 Accepted:2021-04-19 Online:2021-07-01 Published:2021-06-30
Contact: Yuchun Rao,Dan Mu
About author:First author contact:
† These authors contributed equally to this paper

摘要/Abstract

摘要： 舒茶早(Camellia sinensis cv. ‘shuchazao’)是皖西南产区一种茶树品种。为研究茶蚜取食为害诱导的茶树挥发物释放特征, 运用气相色谱-质谱联用技术, 比较了健康茶梢和蚜害茶梢的挥发物组成和相对含量。结果显示, 健康茶梢挥发物种类(16种)和相对含量较少, 而蚜害茶梢挥发物种类(24种)较为丰富且相对含量增大。对具有显著性差异的挥发物种类进行无监督式聚类分析, 结果表明, 健康茶梢和蚜害茶梢挥发物具有明显的聚类特征。基于差异显著挥发物建立了偏最小二乘法判别分析(PLS-DA)模型。经实验验证, 此模型可较好地区分健康和蚜害茶梢(R²X=0.903, R²Y=0.875)。通过计算变量重要性投影, 结果表明, α-蒎烯、长叶烯-(V4)、苯甲醛、反-2-甲基-2-丁烯酸环丙烯酯、3-己烯醛、莰酮和癸醛这7种重要挥发物的相对含量整体变化对判别健康与蚜害茶梢具有重要作用。研究初步揭示了茶蚜为害茶树挥发物含量变化的特征, 可为制定茶蚜防控策略提供新的理论依据。

关键词: 茶树挥发物, 气相色谱-质谱联用技术, PLS-DA, 茶蚜, 聚类分析

Abstract: Shuchazao tea (Camellia sinensis cv. ‘shuchazao’) is a newly cultivated tea species in southwest Anhui, China. To investigate the volatile release profiles of tea aphid (Toxoptera aurantii) infested tea shoots, gas chromatography-mass spectrometry (GC-MS) analysis was used to compare the volatile compositions and relative contents in healthy and infested tea shoots. 16 volatile organic compounds (VOCs) were detected in healthy tea shoots with fewer relative contents, whereas more volatiles (24 VOCs) were released with increasing relative contents in tea plants induced by tea aphid feeding. Unsupervised clustering analysis based on the significantly different volatiles indicated that the volatile profiles in healthy and tea aphid infested tea shoots apparently showed clustering characteristics. A model was created by supervised partial least square discrimination analysis (PLS-DA) for volatiles with statistical significance. By experimental verification, the model could clearly discriminate healthy tea shoots from aphid infested ones (R²X=0.903, R²Y=0.875). By calculating the variable importance for the projection (VIP), seven important volatiles (α-pinene, longifolene-(V4), benaldehyde, E-5,9-undecadien-2-one,6,10-dimethyl-, 3-hexenal, camphor and decanal) were identified which collectively contribute to discrimination between healthy and infested tea shoots. The current work has preliminarily demonstrated the changing patterns in tea plant volatiles after tea aphid infestation to provide novel theoretical guidance for tea aphid management.

Key words: tea plant volatiles, GC-MS, PLS-DA, tea aphid, clustering analysis

孙廷哲, 岂泽华, 梁可欣, 李沁, 饶玉春, 穆丹. 蚜害茶树挥发物组分变化的聚类分析. 植物学报, 2021, 56(4): 422-432.

Tingzhe Sun, Zehua Qi, Kexin Liang, Qin Li, Yuchun Rao, Dan Mu. Clustering Analysis of Volatile Components from the Tea Plants Infested by Tea Aphid (Toxoptera aurantii). Chinese Bulletin of Botany, 2021, 56(4): 422-432.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB21031

https://www.chinbullbotany.com/CN/Y2021/V56/I4/422

图/表 7

表1 健康和蚜害茶梢挥发物组分(VOCs)和相对含量(平均值±标准差)

Table 1 The relative content of volatile organic compounds (VOCs) from healthy and infested tea shoots by Toxoptera aurantii (means±SE)

No.	Retention time (min)	Volatile organic compounds	Relative content
No.	Retention time (min)	Volatile organic compounds	Healthy	Infested
C1	4.842	3-hexenal	0	1.3204±0.3300**
C2	6.984	Ethylbenzene	0.3174±0.0895	0.8110±0.2546
C3	7.391	Benzene,1,3-dimethy-	0.7028±0.0881	1.5338±0.5370
C4	8.261	p-xylene	0.1596±0.0382	0.5888±0.1525
C5	8.572	2-heptanone	0	0.6570±0.2495**
C6	10.057	α-pinene	0	0.3518±0.0670**
C7	11.485	Benaldehyde	0	0.5694±0.1158**
C8	13.402	Decane	0	0.1562±0.0716**
C9	13.658	Octanal	0.0350±0.0080	0.3076±0.1001
C10	14.796	2-ethyl-1-hexanol	0.4416±0.1631	1.1204±0.2834
C11	16.637	Acetophenone	0.0990±0.0240	0.4676±0.1435
C12	18.471	Undecane	0.0756±0.0166	0.1842±0.0535
C13	18.700	Nonanal	0.1240±0.0379	0.5608±0.2060*
C14	19.071	E-2-butenoic acid, 2-(methylencyclopropyl)prop-2-ylester	0	0.0336±0.0123**
C15	20.682	Camphor	0.1174±0.0318	0.5498±0.1094**
C16	21.912	E-2-nonen-1-ol	0	0.1422±0.0804**
C17	22.387	Naphthalene	0.0552±0.0174	0.2874±0.0868*
C18	23.367	Dodecane	0.0334±0.0092	0.1366±0.0350*
C19	23.666	Decanal	0.1188±0.0332	0.6450±0.3569*
C20	28.033	Tridecane	0.0160±0.0036	0.0384±0.0142
C21	32.427	Tetradecane	0.0358±0.0094	0.1904±0.0688*
C22	32.612	Longifolene-(V4)	0	0.0648±0.0141**
C23	34.363	E-5,9-undecadien-2-one,6,10-dimethyl-	0.0470±0.0118	0.2866±0.1340*
C24	40.538	Hexadecane	0.0408±0.0126	0.2320±0.1007*

图1 健康茶梢(A)和蚜害茶梢(B)挥发物组分的GC-MS总离子流 C1-C24同表1。IS: 内标。

Figure 1 The total ion chromatograms of the volatile organic compounds from healthy (A) and infested (B) tea shoots by Toxoptera aurantii C1-C24 are the same as Table 1. IS: Internal standard.

表2 茶树挥发物分类

Table 2 Classification of volatile organic compounds from tea plants

Volatile classification	Volatile compounds
GLVs	3-hexenal and 2-ethyl-1-hexanol
Aromatics	Ethylbenzene, benzene,1,3-dimethy-, p-xylene, benaldehyde, acetophenone and naphthalene
Terpenes	α-pinene, E-5,9-undecadien-2-one,6,10-dimethyl-, E-2-butenoic acid, 2-(methylen-cyclopropyl)prop-2-ylester and longifolene-(V4)
Alkanes	Decane, undecane, dodecane, tridecane, tetradecane and hexadecane
Other	2-heptanone, octanal, nonanal, camphor, E-2- nonen-1-ol and decanal

图2 健康茶梢和蚜害茶梢挥发物种类和比例 (A) 健康茶梢和蚜害茶梢5种挥发物的相对含量(**P<0.01); (B) 健康茶梢和蚜害茶梢5种挥发物占比。GLVs同表2。

Figure 2 The categories and proportions of volatile organic compounds in healthy and infested tea shoots (A) The relative contents of five kinds of volatiles in healthy and infested tea shoots (**P<0.01); (B) The proportion of five volatile compounds in healthy and infested tea shoots. GLVs is the same as given in Table 2.

图3 茶梢挥发物组成二维嵌入和聚类分析 (A) 5组健康茶梢(编号1-5)和5组蚜害茶梢(编号6-10)挥发物组成(VOCs)的t分布随机邻域嵌入(t-SNE); (B) 10组茶梢挥发物组分聚类分析

Figure 3 Two-dimensional embedding and clustering for volatiles from tea shoots (A) The t-distributed stochastic neighbor embedding (t-SNE) for volatile organic compounds (VOCs) from 5 healthy (No. 1-5) and 5 infested (No.6-10) groups; (B) Clustering analysis for VOCs from 10 groups above

图4 茶树挥发物组成(VOCs)主成分分析 (A) 5组健康茶梢(红色点, 因部分重叠故编号未予显示)和5组蚜害茶梢(编号6-10, 蓝色点)挥发物成分的主成分分析得分图; (B) 第1主成分(红色)和第2主成分(蓝色)载荷图(挥发物编号同表1)

Figure 4 Principal component analysis for volatile organic compounds (VOCs) of tea plants (A) The principal component scores for VOCs from 5 healthy (red points, owing to the partial overlap among points in healthy group, the number was not marked) and 5 infested (No.6-10, blue points) groups; (B) Loading plot for the first (red) and second (blue) principal components (the number of volatiles is the same as Table 1)

图5 茶树挥发物组分的偏最小二乘法判别分析 (A) 前2个隐变量(LVs)累计解释总变异百分比(红色为自变量, 蓝色为因变量); (B) 健康组和蚜害组在前2个隐变量的得分图(椭圆为基于Hotelling T2计算出的95%置信域); (C) 偏最小二乘法判别分析双标图(实线圆半径为1.0, 虚线圆半径为0.5); (D) 各挥发物(编号同表1)的变量重要性投影(VIP)图(虚线为取值1.0引导线)

Figure 5 The partial least square discriminant analysis for volatile contents of tea plants (A) The fraction of cumulative explained variations for the first two latent variables (LVs) (red: predictor, blue: response); (B) Scores for the healthy and infested groups with respect to the LV1 and LV2 (the eclipse denotes the 95% confidence interval based on Hotelling T2); (C) The biplot in partial least squares discrimination analysis (the radius for solid and dashed circle is 1.0 and 0.5, respectively); (D) Variable importance for the projection (VIPs) for each volatile (the number is the same as Table 1) organic compound (the dashed line is a guideline for 1.0)

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蚜害茶树挥发物组分变化的聚类分析

Clustering Analysis of Volatile Components from the Tea Plants Infested by Tea Aphid (Toxoptera aurantii)

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