外源物质对茶树耐寒及蔗糖代谢关键基因表达的影响

doi:10.11983/CBB19024

摘要/Abstract

摘要： 冬春季节低温伤害是影响茶树(Camellia sinensis)生产的重要因素。以茶树盆栽苗为试验材料, 通过喷施不同浓度的γ-氨基丁酸(GABA)、绿藻粉和竹醋液溶液, 研究低温胁迫下3种外源物质对茶树耐寒能力的影响, 并对蔗糖代谢关键基因SPS、SUS4、INV4和INV5表达量及耐寒相关生理指标进行分析, 解析外源物质影响茶树耐寒性的生理与分子机制。结果表明, 低温胁迫下, 用不同浓度GABA、绿藻粉和竹醋液喷施处理茶树盆栽苗叶片, 其冻害指数和相对电导率显著低于对照; 可溶性糖含量显著高于对照, 以10 mmol·L ^-1GABA、0.22 mg·mL ^-1绿藻粉及2.5 mg·mL ^-1竹醋液处理效果最佳。低温胁迫下, 与清水处理相比, 分别用10 mmol·L ^-1GABA、0.22 mg·mL ^-1绿藻粉或2.5 mg·mL ^-1竹醋液处理茶树盆栽苗后, 茶树苗丙二醛含量显著降低, 抗氧化酶活性显著提高; 叶片叶绿素、可溶性糖和脯氨酸含量显著增加, 蔗糖含量在处理72小时后分别增加了15.24%、11.39%和5.97%; SPS、SUS4、INV4和INV5基因的表达量显著升高。实验结果表明, GABA、绿藻粉和竹醋液能显著增强茶树的耐寒性。研究结果可为茶树抗寒剂的筛选提供理论依据。

关键词: 茶树, 低温胁迫, 可溶性糖, 蔗糖, 抗寒剂

Abstract: In recent years, low temperature damage in winter and spring has become an important factor affecting tea production. We used tea tree (Camellia sinensis) potted seedlings to test the effect of GABA, chlorella powder and bamboo vinegar on sucrose-related gene expression (SPS, SUS4, INV4, INV5) and cold-tolerant related physiological indexes. Different concentrations of the three kinds of exogenous substances were sprayed on seedlings under cold stress. We analyzed the expression of SPS, SUS4, INV4 and INV5 of the key sucrose-coding genes as well as the physiological indexes related to cold tolerance to determine the possible physiological and molecular mechanisms of the effect of exogenous substance spraying on cold tolerance of tea trees. The frost index and relative conductivity were significantly lower after application of GABA, chlorella powder or bamboo vinegar than control values and the soluble sugar content was increased. The optimal application concentration for GABA, chlorella powder and bamboo vinegar was 10 mmol·L ^-1, 0.22 mg·mL ^-1 and 2.5 mg·mL ^-1, respectively. Malondialdehyde content was significantly decreased and antioxidant enzyme activity was increased; chlorophyll, soluble sugar and proline content was also increased. Sucrose content was increased by 15.24%, 11.39% and 5.97%, respectively, after 72 h treatment. The expression of SPS, SUS4, INV4 and INV5 in leaves was significantly increased with 10 mmol·L ^-1 GABA, 0.22 mg·mL ^-1 chlorella powder and 2.5 mg·mL ^-1 bamboo vinegar. Thus, GABA, chlorophyll powder and bamboo vinegar can significantly improve the cold tolerance of tea plants. We provide a theoretical basis for the cryoprotectant selection for tea plants.

Key words: tea tree, low temperature stress, soluble sugar, sucrose, cryoprotectant

杨小青,黄晓琴,韩晓阳,刘腾飞,岳晓伟,伊冉. 外源物质对茶树耐寒及蔗糖代谢关键基因表达的影响. 植物学报, 2020, 55(1): 21-30.

Xiaoqing Yang,Xiaoqin Huang,Xiaoyang Han,Tengfei Liu,Xiaowei Yue,Ran Yi. Effect of Exogenous Substances on Cold Tolerance and Key Sucrose Metabolic Gene Expression in Camellia sinensis. Chinese Bulletin of Botany, 2020, 55(1): 21-30.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB19024

https://www.chinbullbotany.com/CN/Y2020/V55/I1/21

图/表 8

表1 本研究所用引物

Table 1 Primers used in this study

Primer name	Sequences (5′-3′)
ACTIN-F	GCCATCTTTGATTGGAATGG
ACTIN-R	GGTGCCACAACCTTGATCTT
SPS-F	ACCTGGAGGCGATTCTGGATG
SPS-R	TTCCAAATCCGCCAGCACATA
SUS4-F	TCCTACACTCCGCACCAAGG
SUS4-R	AGCATCTCTTCATGTAGTAA
INV4-F	TGACAAAACAAAGGCTCGACT
INV4-R	CTAAAACACCATAGCCAAGA
INV5-F	CTAAATCTCGATCCTCCTAC
INV5-R	CCAGATAGCAGGAGGACACG

图1 低温胁迫下不同浓度GABA、绿藻粉和竹醋液处理对茶树叶片冻害指数的影响 CK: 清水; T1-1: 5 mmol·L-1 GABA; T1-2: 10 mmol·L-1 GABA; T1-3: 20 mmol·L-1 GABA; T2-1: 0.67 mg·mL-1绿藻粉; T2-2: 0.33 mg·mL-1绿藻粉; T2-3: 0.22 mg·mL-1绿藻粉; T2-4: 0.17 mg·mL-1绿藻粉; T3-1: 5.0 mg·mL-1竹醋液; T3-2: 3.3 mg·mL-1竹醋液; T3-3: 2.5 mg·mL-1竹醋液; T3-4: 2.0 mg·mL-1竹醋液。不同小写字母表示各处理间差异显著(P<0.05)。

Figure 1 The effect of GABA, chlorella powder and bamboo vinegar treatments on cold injury index of tea leaves under low temperature stress CK: Water; T1-1: 5 mmol·L-1 GABA; T1-2: 10 mmol·L-1 GABA; T1-3: 20 mmol·L-1 GABA; T2-1: 0.67 mg·mL-1 chlorella powder; T2-2: 0.33 mg·mL-1 chlorella powder; T2-3: 0.22 mg·mL-1 chlorella powder; T2-4: 0.17 mg·mL-1 chlorella powder; T3-1: 5.0 mg·mL-1 bamboo vinegar; T3-2: 3.3 mg·mL-1 bamboo vinegar; T3-3: 2.5 mg·mL-1 bamboo vinegar; T3-4: 2.0 mg·mL-1 bamboo vinegar. Different lowercase letters indicate significant differences among the treatments (P<0.05).

图2 低温胁迫下不同浓度GABA (A)、绿藻粉(B)和竹醋液(C)处理的茶树叶片相对电导率数据为3次重复的平均值。柱上不同小写字母表示各处理间差异显著(P<0.05)。

Figure 2 The relative conductivity of tea leaves treated with different concentrations of GABA (A), chlorella powder (B) and bamboo vinegar (C) under low temperature stress The data are means of three replicates. Different lowercase letters above the bars indicate significant differences among the treatments (P<0.05).

图3 低温胁迫下不同浓度GABA (A)、绿藻粉(B)和竹醋液(C)处理的茶树叶片可溶性糖含量数据为3次重复的平均值。柱上不同小写字母表示各处理间差异显著(P<0.05)。

Figure 3 The soluble sugar content of tea leaves treated with different concentrations of GABA (A), chlorella powder (B) and bamboo vinegar (C) under low temperature stress The data are means of three replicates. Different lowercase letters above the bars indicate significant differences among the treatments (P<0.05).

图4 低温胁迫下GABA、绿藻粉和竹醋液处理的茶树叶片叶绿素含量数据为3次重复的平均值。柱上不同小写字母表示各处理间差异显著(P<0.05)。

Figure 4 The chlorophyll content of tea leaves treated with GABA, chlorella powder and bamboo vinegar under low temperature stress The data are means of three replicates. Different lowercase letters above the bars indicate significant differences among the treatments (P<0.05).

图5 低温胁迫下GABA、绿藻粉和竹醋液处理的茶树叶片可溶性糖(A)、脯氨酸(B)和蔗糖(C)含量

Figure 5 The soluble sugar (A), proline (B) and sucrose (C) content of tea leaves treated with GABA, chlorella powder and bamboo vinegar under low temperature stress

图6 低温胁迫下GABA、绿藻粉和竹醋液处理的茶树叶片丙二醛(MDA)含量(A)以及过氧化氢酶(CAT) (B)、过氧化物酶(POD) (C)和超氧化物歧化酶(SOD) (D)活性

Figure 6 The malondialdehyde (MDA) content (A) and activities of catalase (CAT) (B), peroxidase (POD) (C), superoxide dismutase (SOD) (D) of tea leaves treated with GABA, chlorella powder and bamboo vinegar under low temperature stress

图7 低温胁迫下GABA、绿藻粉和竹醋液处理的茶树叶片SPS (A)、SUS4 (B)、INV4 (C)和INV5 (D)基因表达量数据为3次重复的平均值。柱上不同小写字母表示各处理间差异显著(P<0.05)。

Figure 7 The effect of GABA, chlorella powder and bamboo vinegar on the expression of SPS (A), SUS4 (B), INV4 (C), and INV5 (D) genes in tea leaves under low temperature stress The data are means of three replicates. Different lowercase letters above the bars indicate significant differences among the treatments (P<0.05).

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