Effect of Exogenous Substances on Cold Tolerance and Key Sucrose Metabolic Gene Expression in Camellia sinensis

doi:10.11983/CBB19024

Abstract

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

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[J]. Chinese Bulletin of Botany, 2020, 55(1): 21-30.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB19024

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

Figures/Tables 8

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

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).

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).

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).

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).

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

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

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).

References 30

[1]	艾沙江·买买提, 张校立, 梅闯, 闫鹏, 王继勋 (2016). 保水剂、绿藻粉处理对库尔勒香梨抗冻能力的影响. 新疆农业科学 53, 445-454.
[2]	陈贵, 胡文玉, 谢甫绨, 张立军 (1991). 提取植物体内MDA的溶剂及MDA作为衰老指标的探讨. 植物生理学通讯 27, 44-46.
[3]	陈杭芳 (2007). NaHSO₃提高春茶产量、品质和抗冻性机制探讨. 硕士论文. 杭州: 浙江大学. pp. 27-35.
[4]	丁菲 (2012). 低温胁迫下与茶树糖代谢相关基因的克隆与表达. 硕士论文. 合肥: 安徽农业大学. pp. 1-8.
[5]	葛菁 (2012). HPLC-ELSD测定茶树可溶性糖含量及GS基因的克隆与表达. 硕士论文. 合肥: 安徽农业大学. pp. 3-5.
[6]	黄娟 (2015). 外源GABA对温度胁迫下黄瓜生长的影响. 硕士论文. 重庆: 西南大学. pp. 25-42.
[7]	黄伟峰, 商晗武, 高永生, 胡剑光, 余秋珠, 王道泽 (2012). 竹醋基液抗寒剂对茶树抗寒性的影响. 茶叶科学 32, 432-440.
[8]	李合生 (1999). 植物生理生化实验原理和技术. 北京: 高等教育出版社. pp. 164-165.
[9]	林郑和, 钟秋生, 游小妹, 陈志辉, 陈常颂, 单睿阳, 阮其春 (2018). 低温胁迫对茶树抗氧化酶活性的影响. 茶叶科学 38, 363-371.
[10]	刘慧英, 朱祝军 (2002). 转化酶在高等植物蔗糖代谢中的作用研究进展. 植物学报 19, 666-674.
[11]	刘凌霄, 沈法富, 卢合全, 韩庆点, 刘云国 (2005). 蔗糖代谢中蔗糖磷酸合成酶(SPS)的研究进展. 分子植物育种 3, 275-281.
[12]	孙鲁龙, 耿庆伟, 邢浩, 杜远鹏, 翟衡 (2017). 低温处理葡萄根系对叶片PSII活性的影响. 植物学报 52, 159-166.
[13]	孙世利, 骆耀平, 潘顺顺, 凌彩金, 苗爱清, 赵超艺 (2008). 壳聚糖对茶树抗寒性相关指标的影响研究. 广东农业科学 ( 12), 34-37.
[14]	田景花, 王红霞, 张志华, 高仪 (2015). 低温逆境对不同核桃品种抗氧化系统及超微结构的影响. 应用生态学报 26, 1320-1326.
[15]	王连荣, 刘铁铮 (2010). 果树抗寒生理与防寒措施的研究进展. 安徽农业科学 38, 9483-9484.
[16]	王平盛, 虞富莲 (2002). 中国野生大茶树的地理分布、多样性及其利用价值. 茶业科学 22, 105-108, 134.
[17]	王素平, 郭世荣, 胡晓辉, 李璟, 焦彦生 (2006). 盐胁迫对黄瓜幼苗叶片光合色素含量的影响. 江西农业大学学报 28, 32-38.
[18]	王育平 (2014). 茶树冻害成因及预防补救措施. 福建农业科技 ( 5), 47-49.
[19]	熊先军, 刘明月 (2005). 辣椒抗寒性生理生化研究进展. 辣椒杂志 1, 9-12.
[20]	许英, 陈建华, 朱爱国, 栾明宝, 王晓飞, 孙志民 (2015). 低温胁迫下植物响应机理的研究进展. 中国麻业科学 37, 40-49.
[21]	岳川 (2015). 茶树糖类相关基因的挖掘及其在茶树冷驯化中的表达研究. 博士论文. 北京: 中国农业科学院. pp. 52-75.
[22]	张贱根 (2006). 茶树冻害的发生与预防补救措施. 茶业通报 28(1), 12-13.
[23]	张丽, 周欣, 蒋家月, 江昌俊, 朱政 (2012). 外源ABA对茶树抗寒生理指标的影响. 茶业通报 34, 72-74.
[24]	周琳, 陈暄, 王玉花, 朱旭君, 黎星辉, 房婉萍 (2014). 超敏蛋白对低温胁迫下茶树生理特性的影响. 园艺学报 41, 746-754.
[25]	Baud S, Vaultier MN, Rochat C (2004). Structure and expression profile of the sucrose synthase multigene family in Arabidopsis. J Exp Bot 55, 397-409.
[26]	Benina M, Obata T, Mehterov N, Ivanov I, Petrov V, Toneva V, Fernie AR, Gechev TS (2013). Comparative metabolic profiling of Haberlea rhodopensis, Thellungiella halophyla, and Arabidopsis thaliana exposed to low temperature. Front Plant Sci 4, 499.
[27]	Dhindsa RS, Matowe W (1981). Drought tolerance in two mosses: correlated with enzymatic defence against lipid peroxidation. J Exp Bot 32, 79-91.
[28]	Dong HZ, Sun LH, Jian LC (1980). Ultrastructural changes in leaf cells of wheat varieties with different cold resistance after freezing-thawing of the plants. Acta Bot Sin 22, 339-342.
[29]	Otting G, Liepinsh E, Wuthrich K (1991). Protein hydration in aqueous solution. Science 254, 974-980.
[30]	Ruan YL (2014). Sucrose metabolism: gateway to diverse carbon use and sugar signaling. Ann Rev Plant Biol 65, 33-67.