外源亚精胺对盐胁迫下茶树光合特性及关键酶基因表达的影响

doi:10.16420/j.issn.0513-353x.2020-0946

摘要/Abstract

摘要：

为了探讨外源亚精胺（Spd）对盐胁迫下茶树光合特性的调控机理,以茶树品种‘中茶108’为试验材料,通过Spd不同浓度（0、0.5、1.0、1.5 mmol · L^-1）的喷施处理,研究盐胁迫（200 mmol · L^-1 NaCl）下外源Spd对茶树的光合参数、叶绿素含量、气孔开度及关键酶基因表达的影响。结果显示：盐胁迫处理显著降低了茶树叶片的净光合速率（P_n）、蒸腾速率（T_r）及叶绿素含量等,提高了叶片的胞间CO₂浓度（C_i）,处理初期（1、3 d）促进气孔开度,后期（5、9 d）逐渐抑制气孔开度;盐胁迫下,外源Spd显著提高了茶树的P_n、T_r、气孔导度（G_s）、叶绿素a、b及总含量;外源Spd可以不同程度地诱导CsRbcL、CsTK、CsFBPase、CsPRK等光合碳同化基因的表达,并且缓解了盐胁迫对大多叶绿素合成基因抑制的影响。研究表明叶面喷施外源Spd可以有效缓解叶绿素的降解,促进茶树的光合作用,提高茶树的耐盐性,其中以喷施0.5 mmol · L^-1亚精胺的缓解效应最为显著。

关键词: 茶树, 亚精胺, 盐胁迫, 气孔开度, 光合特性

Abstract:

In order to explore the regulatory effects of exogenous spermidine（Spd）on the photosynthetic characteristics under salt stress of tea plant,tea variety‘Zhongcha 108’was used as the experimental material in this study. Through different concentrations（0,0.5,1.0,1.5 mmol · L^-1）Spd spray treatments,the effects of exogenous Spd on tea photosynthetic parameters,chlorophyll content,stomatal aperture and the expression levels of key enzyme genes under salt stress（200 mmol · L^-1NaCl）were detected and analyzed. The results showed that the net photosynthetic rate（P_n）,transpiration rate（T_r）,and chlorophyll content were significantly reduced,and the intercellular CO₂concentration（C_i）was increased under salt stress. Stomatal aperture was promoted the initial treatment（1,3 d）,and inhibited gradually in the later period（5,9 d）. After spraying exogenous Spd,P_n,T_r,stomatal conductance（G_s）,chlorophyll a,chlorophyll b and Chlorophyll（a + b）contents were significantly increased under salt stress. Exogenous Spd can induce the up-regulation of CsRbcL,CsTK,CsFBPase,CsPRK and other photosynthetic carbon assimilation genes to varying degrees,and alleviate the influence of salt stress on the inhibition most chlorophyll synthesis genes. The study demonstrated that spraying exogenous Spd can effectively alleviate the degradation of chlorophyll,promote photosynthesis,and improve the salt tolerance of tea plant,of which 0.5 mmol · L^-1Spd was the most effective.

Key words: tea plant, spermidine, salt stress, stomatal aperture, photosynthetic characteristics

中图分类号:

S571.1

杨妮, 万绮雯, 李逸民, 韩妙华, 滕瑞敏, 刘洁霞, 庄静. 外源亚精胺对盐胁迫下茶树光合特性及关键酶基因表达的影响[J]. 园艺学报, 2022, 49(2): 378-394.

YANG Ni, WAN Qiwen, LI Yimin, HAN Miaohua, TENG Ruimin, LIU Jiexia, ZHUANG Jing. Effects of Exogenous Spermidine on Photosynthetic Characteristics and Gene Expression of Key Enzymes Under Salt Stress in Tea Plant[J]. Acta Horticulturae Sinica, 2022, 49(2): 378-394.

图/表 8

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基因 Gene	名称 Name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
CsACT7	肌动蛋白 Actin	TTGGATTCTGGGGATGGTGTTAGC	AGCAAGTTTCTCTTTCACATCACGG
CsADC	精氨酸脱酸酶 Arginine decarboxylase	TCAAGGAAGGGTCTTTGGGTA	GGTATGTGCGAATGGGGTCT
CsCHLM	Mg-原卟啉 Ⅸ 甲基转移酶 Magnesium protoporphyrin Ⅸ methyltransferase	CTCTATTGCCTCATTCCTC	ATTTAGTGTTTGGGTTGGT
CsCRD1	Mg-原卟啉 Ⅸ 单甲基酯环化酶 Magnesium protoporphyrin Ⅸ monomethyl ester cyclase	CAATGACTGGAAGGCTAA	ATTCTTTGGTGTTGAGGC
CsFBPase	果糖-1,6-二磷酸酶 Furetose-1,6-bisphosphate phosphatase	GCAGATTGCTTCGTTGGTTC	TGCTATTATCCCTGTCCTCCC
CsHEMA1	谷氨酸-tRNA还原酶 Glutamyl-tRNA reductase	ATTCGTTGCGAGATTGTT	GCTGCTCCTTTCCTTTGT
CsHEMC	胆色素原脱氨酶 Porphobilinogen deaminase	TGACCGCCATTCTTTCTA	GCTAATCTTGTTTCCTCGT
CsHEMD	尿卟啉原 Ⅲ 合成酶 Uroporphyrinogen Ⅲ synthase	TGTCTGGGCTGTCTTCGA	CAAATCAGGCAACCGTGT
CsHEME2	尿卟啉原 Ⅲ 脱羧酶 Uroporphyrinogen Ⅲ decarboxylase	ACATTCGCTTCTGTTCCC	TTTCTACTTCCAGCCCTC
CsHEMG2	原卟啉原氧化酶 Protoporphyrinogen oxidase	TCTGTGGAAGAAACGGAACT	CCGCAACGAAAGGGTCAA
CsODC	鸟氨酸脱羧酶 Ornithine decarboxylase	TTCCAAACATGGGTGCGTAT	TGTGAGGTGGGTCGTAATCG
CsPRK	核酮糖-5-磷酸激酶 Ribulose5-phosphate kinase	AGTATTGGAGCCCGAAAGCC	CAAACAGGTAAACTGGGGTGAA
CsPPOX	原卟啉原氧化酶 Protoporphyrinogen oxidase	GAAGCAGTTGACCGTGAC	AAAGCCCTCTGAACCCAC
CsRbcL	核酮糖-1,5-二磷酸羧化/加氧酶大亚基 Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit	TGGCTGGAGATGGGACGA	CCTCTGGTAATCAGAACAGGGTT
CsSAMDC	S-腺苷甲硫氨酸脱羧酶 S-adenosylmethionine decarboxylase	TACTCTGCTTCTGCTGAATCGA	GCCTTCTTCCTGTCCAAACC
CsSPDS	亚精胺合成酶 Spermidine synthases	AACCCAATAGACGCCGATGA	CGCAAAAGATGGCAAACAGA
CsSAMS	S-腺苷甲硫氨酸合成酶 S-adenosylmethionine synthases	CGATGAGACCGTGACAAATGA	AGGGTTGAGGTGGAAGATGG
CsTK	转酮醇酶 Transketolase	TGCCCAATGTTTTGATGCTACG	ATCCACCCTTTTCCACTCCCTC

基因 Gene	名称 Name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
CsACT7	肌动蛋白 Actin	TTGGATTCTGGGGATGGTGTTAGC	AGCAAGTTTCTCTTTCACATCACGG
CsADC	精氨酸脱酸酶 Arginine decarboxylase	TCAAGGAAGGGTCTTTGGGTA	GGTATGTGCGAATGGGGTCT
CsCHLM	Mg-原卟啉 Ⅸ 甲基转移酶 Magnesium protoporphyrin Ⅸ methyltransferase	CTCTATTGCCTCATTCCTC	ATTTAGTGTTTGGGTTGGT
CsCRD1	Mg-原卟啉 Ⅸ 单甲基酯环化酶 Magnesium protoporphyrin Ⅸ monomethyl ester cyclase	CAATGACTGGAAGGCTAA	ATTCTTTGGTGTTGAGGC
CsFBPase	果糖-1,6-二磷酸酶 Furetose-1,6-bisphosphate phosphatase	GCAGATTGCTTCGTTGGTTC	TGCTATTATCCCTGTCCTCCC
CsHEMA1	谷氨酸-tRNA还原酶 Glutamyl-tRNA reductase	ATTCGTTGCGAGATTGTT	GCTGCTCCTTTCCTTTGT
CsHEMC	胆色素原脱氨酶 Porphobilinogen deaminase	TGACCGCCATTCTTTCTA	GCTAATCTTGTTTCCTCGT
CsHEMD	尿卟啉原 Ⅲ 合成酶 Uroporphyrinogen Ⅲ synthase	TGTCTGGGCTGTCTTCGA	CAAATCAGGCAACCGTGT
CsHEME2	尿卟啉原 Ⅲ 脱羧酶 Uroporphyrinogen Ⅲ decarboxylase	ACATTCGCTTCTGTTCCC	TTTCTACTTCCAGCCCTC
CsHEMG2	原卟啉原氧化酶 Protoporphyrinogen oxidase	TCTGTGGAAGAAACGGAACT	CCGCAACGAAAGGGTCAA
CsODC	鸟氨酸脱羧酶 Ornithine decarboxylase	TTCCAAACATGGGTGCGTAT	TGTGAGGTGGGTCGTAATCG
CsPRK	核酮糖-5-磷酸激酶 Ribulose5-phosphate kinase	AGTATTGGAGCCCGAAAGCC	CAAACAGGTAAACTGGGGTGAA
CsPPOX	原卟啉原氧化酶 Protoporphyrinogen oxidase	GAAGCAGTTGACCGTGAC	AAAGCCCTCTGAACCCAC
CsRbcL	核酮糖-1,5-二磷酸羧化/加氧酶大亚基 Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit	TGGCTGGAGATGGGACGA	CCTCTGGTAATCAGAACAGGGTT
CsSAMDC	S-腺苷甲硫氨酸脱羧酶 S-adenosylmethionine decarboxylase	TACTCTGCTTCTGCTGAATCGA	GCCTTCTTCCTGTCCAAACC
CsSPDS	亚精胺合成酶 Spermidine synthases	AACCCAATAGACGCCGATGA	CGCAAAAGATGGCAAACAGA
CsSAMS	S-腺苷甲硫氨酸合成酶 S-adenosylmethionine synthases	CGATGAGACCGTGACAAATGA	AGGGTTGAGGTGGAAGATGG
CsTK	转酮醇酶 Transketolase	TGCCCAATGTTTTGATGCTACG	ATCCACCCTTTTCCACTCCCTC

处理后天数/d Days after treatment	处理/（mmol · L^-1） Treatment		气孔横径/μm Stomatal transverse diameter	气孔纵径/μm Stomatal longitudinal diameter	气孔开度/μm² Stomatal aperture
处理后天数/d Days after treatment	NaCl	Spd	气孔横径/μm Stomatal transverse diameter	气孔纵径/μm Stomatal longitudinal diameter	气孔开度/μm² Stomatal aperture
1	0（对照Control）	0	8.75 ± 0.13 lm	17.81 ± 0.71 def	122.54 ± 6.67 jk
	0	0.5	11.88 ± 0.16 abcde	20.21 ± 0.41 abcd	188.49 ± 6.36 b
	0	1.0	9.60 ± 0.29 ijklm	18.24 ± 0.73 bcdef	137.16 ± 2.38 hi
	0	1.5	12.51 ± 0.22 a	18.79 ± 0.20 abcde	184.48 ± 1.27 bc
	200	0	10.44 ± 0.31 efghijk	18.83 ± 0.46 abcde	154.03 ± 1.55 fg
	200	0.5	8.69 ± 0.19 lm	17.15 ± 0.29 ef	116.85 ± 1.28 k
	200	1.0	10.87 ± 0.32 cdefghi	16.11 ± 0.21 f	137.47 ± 4.17 hi
	200	1.5	9.88 ± 0.29 hijkl	18.85 ± 0.74 abcde	145.80 ± 2.59 gh
3	0（对照Control）	0	10.39 ± 0.01 efghijk	17.61 ± 0.38 def	143.64 ± 3.33 gh
	0	0.5	10.05 ± 0.09 ghijkl	19.25 ± 0.45 abcde	151.90 ± 2.27 g
	0	1.0	11.10 ± 0.27 abcdefgh	19.64 ± 0.85 abcde	171.00 ± 3.29 de
	0	1.5	12.32 ± 0.62 abc	19.54 ± 0.60 abcde	188.59 ± 6.56 b
	200	0	11.74 ± 0.55 abcde	19.16 ± 0.99 abcde	175.68 ± 1.93 bcde
	200	0.5	9.11 ± 0.35 klm	21.08 ± 0.48 a	150.63 ± 2.34 g
	200	1.0	9.09 ± 0.15 klm	17.53 ± 0.48 def	125.04 ± 1.40 ijk
	200	1.5	9.81 ± 0.66 hijkl	17.91 ± 1.08 cdef	136.88 ± 1.09 hi
5	0（对照Control）	0	10.92 ± 0.41 bcdefghi	20.09 ± 1.14 abcd	171.96 ± 8.84 cde
	0	0.5	11.16 ± 0.68 abcdefgh	20.54 ± 0.47 abc	179.77 ± 6.74 bcd
	0	1.0	10.64 ± 0.53 defghij	20.17 ± 0.70 abcd	167.52 ± 2.31 de
	0	1.5	12.39 ± 0.25 ab	21.29 ± 0.35 a	206.93 ± 1.20 a
	200	0	10.98 ± 0.32 bcdefghi	19.89 ± 0.59 abcd	171.24 ± 0.81 de
	200	0.5	10.72 ± 0.56 defghij	20.99 ± 1.62 a	175.85 ± 4.47 bcde
	200	1.0	11.55 ± 0.16 abcdef	19.15 ± 0.17 abcde	173.63 ± 4.02 cde
	200	1.5	11.53 ± 0.63 abcdefg	19.09 ± 1.07 abcde	171.28 ± 3.86 de
9	0（对照Control）	0	11.50 ± 0.21 abcdefg	18.25 ± 0.34 bcdef	164.74 ± 3.18 ef
	0	0.5	9.27 ± 0.47 jklm	17.69 ± 1.03 def	128.15 ± 5.21 ijk
	0	1.0	12.27 ± 0.33 abc	19.54 ± 0.74 abcde	187.86 ± 2.88 b
	0	1.5	11.79 ± 0.08 abcde	18.78 ± 0.51 abcde	173.70 ± 3.57 cde
	200	0	8.28 ± 0.41 m	20.08 ± 0.73 abcd	129.99 ± 1.68 ij
	200	0.5	11.56 ± 0.34 abcdef	20.72 ± 0.45 ab	187.98 ± 1.47 b
	200	1.0	11.97 ± 0.39 abcd	19.10 ± 0.59 abcde	178.85 ± 2.61 bcd
	200	1.5	10.13 ± 0.22 fghjkl	18.94 ± 0.82 abcde	150.29 ± 3.32 g

处理后天数/d Days after treatment	处理/（mmol · L^-1） Treatment		气孔横径/μm Stomatal transverse diameter	气孔纵径/μm Stomatal longitudinal diameter	气孔开度/μm² Stomatal aperture
处理后天数/d Days after treatment	NaCl	Spd	气孔横径/μm Stomatal transverse diameter	气孔纵径/μm Stomatal longitudinal diameter	气孔开度/μm² Stomatal aperture
1	0（对照Control）	0	8.75 ± 0.13 lm	17.81 ± 0.71 def	122.54 ± 6.67 jk
	0	0.5	11.88 ± 0.16 abcde	20.21 ± 0.41 abcd	188.49 ± 6.36 b
	0	1.0	9.60 ± 0.29 ijklm	18.24 ± 0.73 bcdef	137.16 ± 2.38 hi
	0	1.5	12.51 ± 0.22 a	18.79 ± 0.20 abcde	184.48 ± 1.27 bc
	200	0	10.44 ± 0.31 efghijk	18.83 ± 0.46 abcde	154.03 ± 1.55 fg
	200	0.5	8.69 ± 0.19 lm	17.15 ± 0.29 ef	116.85 ± 1.28 k
	200	1.0	10.87 ± 0.32 cdefghi	16.11 ± 0.21 f	137.47 ± 4.17 hi
	200	1.5	9.88 ± 0.29 hijkl	18.85 ± 0.74 abcde	145.80 ± 2.59 gh
3	0（对照Control）	0	10.39 ± 0.01 efghijk	17.61 ± 0.38 def	143.64 ± 3.33 gh
	0	0.5	10.05 ± 0.09 ghijkl	19.25 ± 0.45 abcde	151.90 ± 2.27 g
	0	1.0	11.10 ± 0.27 abcdefgh	19.64 ± 0.85 abcde	171.00 ± 3.29 de
	0	1.5	12.32 ± 0.62 abc	19.54 ± 0.60 abcde	188.59 ± 6.56 b
	200	0	11.74 ± 0.55 abcde	19.16 ± 0.99 abcde	175.68 ± 1.93 bcde
	200	0.5	9.11 ± 0.35 klm	21.08 ± 0.48 a	150.63 ± 2.34 g
	200	1.0	9.09 ± 0.15 klm	17.53 ± 0.48 def	125.04 ± 1.40 ijk
	200	1.5	9.81 ± 0.66 hijkl	17.91 ± 1.08 cdef	136.88 ± 1.09 hi
5	0（对照Control）	0	10.92 ± 0.41 bcdefghi	20.09 ± 1.14 abcd	171.96 ± 8.84 cde
	0	0.5	11.16 ± 0.68 abcdefgh	20.54 ± 0.47 abc	179.77 ± 6.74 bcd
	0	1.0	10.64 ± 0.53 defghij	20.17 ± 0.70 abcd	167.52 ± 2.31 de
	0	1.5	12.39 ± 0.25 ab	21.29 ± 0.35 a	206.93 ± 1.20 a
	200	0	10.98 ± 0.32 bcdefghi	19.89 ± 0.59 abcd	171.24 ± 0.81 de
	200	0.5	10.72 ± 0.56 defghij	20.99 ± 1.62 a	175.85 ± 4.47 bcde
	200	1.0	11.55 ± 0.16 abcdef	19.15 ± 0.17 abcde	173.63 ± 4.02 cde
	200	1.5	11.53 ± 0.63 abcdefg	19.09 ± 1.07 abcde	171.28 ± 3.86 de
9	0（对照Control）	0	11.50 ± 0.21 abcdefg	18.25 ± 0.34 bcdef	164.74 ± 3.18 ef
	0	0.5	9.27 ± 0.47 jklm	17.69 ± 1.03 def	128.15 ± 5.21 ijk
	0	1.0	12.27 ± 0.33 abc	19.54 ± 0.74 abcde	187.86 ± 2.88 b
	0	1.5	11.79 ± 0.08 abcde	18.78 ± 0.51 abcde	173.70 ± 3.57 cde
	200	0	8.28 ± 0.41 m	20.08 ± 0.73 abcd	129.99 ± 1.68 ij
	200	0.5	11.56 ± 0.34 abcdef	20.72 ± 0.45 ab	187.98 ± 1.47 b
	200	1.0	11.97 ± 0.39 abcd	19.10 ± 0.59 abcde	178.85 ± 2.61 bcd
	200	1.5	10.13 ± 0.22 fghjkl	18.94 ± 0.82 abcde	150.29 ± 3.32 g