基因转录介导同质园条件下拟南芥不同地理种群的光合能力分化

doi:10.7525/j.issn.1673-5102.2023.01.010

摘要/Abstract

摘要：

不同环境条件决定着植物光合能力的多态性，但在相同环境条件下同种植物不同种群间表现出光合能力分化的内在机制仍不清楚，本文旨在揭示同质园条件下欧洲拟南芥（Arabidopsis thaliana）不同地理种群光合能力的分化以及其基因转录调控机制。在同质园条件下，测定来自欧洲不同气候区的23个拟南芥的地理种群的气体交换参数、叶绿素荧光参数及SPAD值综合比较其光合能力差异。另外，根据测定结果选取光合能力差异的典型种群，利用实时定量PCR技术对光合相关基因表达水平进行验证。比较研究发现欧洲不同气候区拟南芥的地理种群间的气体交换参数差异较大，其中净光合速率的变化范围为2~11 μmol·m^-2·s^-1；而叶绿素荧光参数变异幅度较小，变异幅度几乎不超过10%。聚类分析表明23个拟南芥种群被分为强光合能力和弱光合能力2组，强光合种群主要分布在中欧和西欧地区，净光合速率平均为7.37 μmol·m^-2·s^-1；而弱光合种群则主要分布在东欧和南欧，净光合速率平均为4.46 μmol·m^-2·s^-1。相关性分析发现净光合速率和水分利用效率及SPAD存在显著相关性。冗余分析（RDA）结果表明拟南芥光合能力的分化可能与其所在地区生长季温度与降雨量等环境因素有关。实时定量PCR试验表明，强光合能力的典型种群En-D和Stw-0的PSⅡ和Rubisco相关基因表达水平显著高于弱光合能力的典型种群Wa-1和Per-1，暗示着PSⅡ和Rubisco相关基因的转录差异可能参与了拟南芥种群光合能力分化的调控。本研究明确了同质园中不同拟南芥地理种群的光合能力存在差异，这种差异可能与原产地环境有关，并在长期进化中遗传给了后代。而PSⅡ和Rubisco相关基因转录调控可能参与了拟南芥光合能力的分化。

关键词: 光合能力, 同质园, PSⅡ基因, Rubisco基因, 基因表达

Abstract:

Photosynthesis is a basic metabolism process for plants and provides the material basis for plant growth and development. Different environmental conditions determine the polymorphism of plant photosynthetic capacity， but the mechanisms regulating the divergence of photosynthetic capacity among different populations under the same environmental conditions remain still unknown. This study aims to reveal the divergence of photosynthetic capacity in different geographical populations of Arabidopsis thaliana in Europe under common garden conditions and the mechanisms of their gene transcriptional regulation. A comparative study of photosynthetic characteristics of 23 geographic A. thaliana（Arabidopsis） populations from different regions was conducted under common garden conditions by determining the gas exchange parameters， chlorophyll fluorescence parameters and SPAD values. In addition， the photosynthetic-related gene expressions of the typical populations with differences in photosynthetic capacity was examined using real-time quantitative PCR（RT-qPCR）. The comparative result revealed that gas exchange parameters differed significantly among geographical populations of Arabidopsis from different climatic zones in Europe. The variation range of net photosynthetic rate was 2-11 μmol·m^-2·s^-1.while chlorophyll fluorescence parameters varied to a lesser extent， the variation range was almost no more than 10%. Cluster analysis showed that the 23 Arabidopsis populations were divided into two groups， strong and weak photosynthetic ability respectively. The populations with strong photosynthetic capacity were mainly distributed in central and western Europe， and the average net photosynthetic rate was 7.37 μmol·m^-2·s^-1. The populations with weak photosynthetic capacity were mainly distributed in eastern and southern Europe with an average net photosynthetic rate of 4.46 μmol·m^-2·s^-1. Correlation analysis revealed a significant correlation between net photosynthetic rate， water use efficiency and SPAD. The results of redundancy analysis（RDA） suggested that the divergence of photosynthetic capacity in Arabidopsis might be related to environmental factors such as temperature and rainfall during the growing season in the region. RT-qPCR results showed that the expression of PSⅡ- and Rubisco-related genes were significantly higher in En-D and Stw-0， typical populations with strong photosynthetic capacity， than in Wa-1 and Per-1， typical populations with weak photosynthetic capacity， suggesting that transcriptional differences in PSⅡ and Rubisco genes could be involved in the regulation of photosynthetic capacity differentiation in populations. The results clarified that there were differences in photosynthetic capacity between geographic populations of Arabidopsis under common gardens， and such differences might be related to the environment of origin and have been inherited to future generations during long-term evolution. In contrast， the transcriptional regulation of PSⅡ and Rubisco-related genes might be involved in the differentiation of photosynthetic capacity in Arabidopsis.

Key words: Photosynthesis, common garden conditions, PSⅡ-related gene, rubisco gene, gene expression

中图分类号:

Q945.3

李梦烁, 刘莹泽, 鲁焕, 强胜. 基因转录介导同质园条件下拟南芥不同地理种群的光合能力分化[J]. 植物研究, 2023, 43(1): 90-99.

Mengshuo LI, Yingze LIU, Huan LU, Sheng QIANG. Photosynthetic Capacity Differentiation and Gene Transcription in Different Geographical Populations of Arabidopsis thaliana under Common Garden conditions[J]. Bulletin of Botanical Research, 2023, 43(1): 90-99.

图/表 8

表1

RT-qPCR的引物对信息

基因 Gene	基因ID Gene ID	引物（5′→3′） Primer（5′→3′）	扩增产物大小 Size of amplicon /bp	注释 Annotation
PsbP	AT1G06680	F：5'-CCTCCTAGGGAAACAAGCTTAC-3′ R：5′-CCTGAGATGATGACTCCAGAATG-3′	79	光系统Ⅱ（PSⅡ）的亚单位 Subunits of Photosystem Ⅱ（PSⅡ）
CP22	AT1G44575	F：5′-ATTACCGGGAAAGGAGCATTAG-3′ R：5′-ATGGCAGCGAAGAAGAAGAA-3′	110	PSⅡ相关的色素结合蛋白 PSⅡ-related pigment-binding protein
CAB1	AT1G29930	F：5′-GAAGGTGAAGGAGCTCAAGAA-3′ R：5′-ATGGTCAGCAAGGTTCTCTATC-3′	109	捕光复合体Ⅱ（LHCⅡ）的亚单位 Subunit of light-harvesting complex Ⅱ（LHCⅡ）
LHCB6	AT1G15820	F：5′-ATACCGGCGATCAGGGATA-3′ R：5′-TTCTCAAAGTCCGGCTCATAAA-3′	97	PSII的捕光复合体LHCⅡ The light-harvesting complex LHCⅡ of PSⅡ
Psa2	AT2G34860	F：5′-CCTGCTTCTCCGTTTCTATCTG-3′ R：5′-TGGGTAGTCTCTGAGGAAGAAT-3′	119	调节光系统Ⅰ（PSⅠ）的组装 Regulates the assembly of Photosystem Ⅰ（PSⅠ）
LHCA6	AT1G19150	F：5′-CCGGTGGAAGATTAATGAGAGAG-3′ R：5′-GAGCTACCAGGGAACCATAAAG-3′	112	PSⅠ外周天线蛋白 PSⅠ peripheral antenna protein
CBR	AT5G17770	F：5′-CATTTACGCCAACGTCACATAC-3′ R：5′-CAACACCACCATCCCATACT-3′	134	编码细胞色素还原酶 Encodes cytochrome reductase
RbcS1A	AT1G67090	F：5′-AATTTCCGGACTTAACGTTTGTTT-3′ R：5′-CATCAGACAGTTGAGAATCCGATAGA-3′	69	编码Rubisco小亚基 Encodes the Rubisco small subunit
RbcS1B	AT5G38430	F：5′-GCCAAAGTGAAAAAACTGAAGGTT-3′ R：5′-AAGAGCAGAAATGAAGTGATATGAATAGA-3′	83	编码Rubisco小亚基 Encodes the Rubisco small subunit
RbcS2B	AT5G38420	F：5′-ACCCATTTCTATGTGGTCAATGC-3′ R：5′-TTCACTTTCAAACAATAGTTCCTCAAC-3′	80	编码Rubisco小亚基 Encodes the Rubisco small subunit
RbcS3B	AT5G38410	F：5′-CCTATTGTCTGTGTTCTTTTTCTCTTTATG-3′ R：5′-TCAAGACGCACGGATATATAAATTACA-3′	99	编码Rubisco小亚基 Encodes the Rubisco small subunit
SBP	AT3G55800	F：5′-GGGAAATGGCAGCATGTAAAG-3′ R：5′-TGTATTCGGAGTTGTCGAATGT-3′	97	编码景天庚-1，7-二磷酸酶 Encodes Sedoheptulose-1，7-bisphosphatase
FBP	AT1G43670	F：5′-ATGGTGGCTGCAGGTTATT-3′ R：5′-GAGATGGGTCCAGTGTAAATCC-3′	94	编码果糖-1，6-二磷酸酶 Encodes Fructose-1，6-bisphosphatase
RCA	AT2G39730	F：5′-GGGTATGGTTGACTCTGTCTTC-3′ R：5′-AGGCCTTGGCTAACGTATTC-3′	90	Rubisco活化酶，激活Rubisco Rubisco Activase，activates Rubisco

表1

表2

表3

图1

表4

图2

图3

图4

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种群 Population	北纬度 Latitude /（°）	东经度 Longitude /（°）	净光合速率 Net photosynthetic rate /（μmol·m^-2·s^-1）	蒸腾速率 Transpiration rate /（mmol·m^-2·s^-1）	水分利用效率 Water use efficiency /（μmol·mol^-1）
变异系数 Coefficient of variation	—	—	40.30%	12.79%	40.61%
Aa-0	50.92	9.57	2.38±0.27g	6.27±0.05a	0.38±0.04i
Ak-1	48.07	7.63	4.78±0.32f	5.25±0.14de	0.90±0.05eh
Boot-1	54.49	-3.27	9.65±0.31ab	5.85±0.10ab	1.65±0.05bc
Bs-1	47.51	7.50	6.77±0.48cde	4.91±0.04f	1.37±0.09c
Bsch-0	50.02	8.67	8.94±0.25bc	5.55±0.04bcd	1.62±0.05bc
Ca-0	50.30	8.27	7.28±0.22bc	5.17±0.04def	1.41±0.04c
Col-0	38.30	-92.30	4.30±0.16f	5.54±0.06bcd	0.78±0.03hg
Ema-1	51.30	0.50	4.30±0.38f	3.94±0.04g	1.10±0.10cde
En-D	50.01	8.50	7.48±0.18bc	5.09±0.03def	1.47±0.03c
Hey-1	51.25	5.91	4.11±0.19f	6.12±0.04ab	0.67±0.03hg
Hs-0	52.25	9.44	11.03±027a	5.52±0.11bcd	2.00±0.05a
Kelsterbach-4	50.07	8.53	3.25±0.16fg	5.29±0.05de	0.62±0.03hg
Kz-9	49.50	73.10	5.29±0.22ef	3.77±0.17gh	1.45±0.09c
Ms-0	55.75	37.63	4.28±0.22f	5.20±0.05de	0.83±0.05h
Nw-0	50.50	8.50	6.36±0.44cde	3.40±0.09h	1.89±0.13ab
Per-1	58.00	56.32	3.14±0.38fg	5.60±0.08bcd	0.56±0.06g
Rd-0	50.50	8.50	3.17±0.24fg	5.04±0.26def	0.65±0.04hg
Se-0	38.33	-3.53	8.01±0.41bc	5.77±0.07b	1.40±0.08c
Stw-0	52.03	36.10	6.92±0.25cde	5.52±0.03bcd	1.25±0.04cd
Tscha-1	47.08	9.90	10.73±0.21a	4.98±0.04ef	2.16±0.05a
Tu-0	45.00	7.50	6.83±0.50cde	5.60±0.03bcd	1.22±0.09cde
Ty-0	56.43	-5.23	6.70±0.53cde	5.03±0.05ef	1.33±0.10c
Wa-1	52.30	21.00	4.64±0.42f	5.42±0.11bcd	0.87±0.08h

光合参数 photosynthetic parameter	P_n	T_r	W_UE	S_PAD	Y_Ⅱ	Y_NPQ	Y_NO	Q_NP	Q_N	Q_P	Q_L	R_ET
P_n	1
T_r	0.006	1
W_UE	0.917^**	-0.373	1
S_PAD	0.617^**	0.106	0.520^*	1
Y_Ⅱ	0.218	-0.138	0.253	0.255	1
Y_NPQ	-0.245	0.039	-0.226	-0.218	0.046	1
Y_NO	0.163	-0.163	0.194	0.356	0.396	-0.168	1
Q_NP	-0.391	0.254	-0.436^*	-0.392	-0.400	0.667^**	-0.509*	1
Q_N	-0.336	0.178	-0.346	-0.418^*	-0.363	0.740^**	-0.542^**	0.955**	1
Q_P	-0.079	0.192	-0.110	-0.314	-0.536^**	-0.109	-0.775^**	0.500*	0.525^*	1
Q_L	-0.153	0.212	-0.178	-0.386	-0.532^**	-0.010	-0.654^**	0.599^**	0.623^**	0.963^**	1
R_ET	0.152	0.093	0.113	-0.071	-0.421^*	-0.375	-0.584^**	0.192	0.173	0.817^**	0.705^**	1

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