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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (1): 99-107.doi: 10.3724/SP.J.1006.2022.04275

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

NtPHYB1 interacts with light and temperature signal to regulate seed germination in Nicotiana tabacum L.

LI Zhen-Hua1,*(), WANG Xian-Ya1, LIU Yi-Ling2, ZHAO Jie-Hong3   

  1. 1College of Agriculture, Guizhou University, Guiyang 550025, Guizhou, China
    2Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou University, Guiyang 550025, Guizhou, China
    3CNTC Key Laboratory of Molecular Genetics, Guizhou Academy of Tobacco Science, Guiyang 550081, Guizhou, China
  • Received:2020-12-15 Accepted:2021-04-14 Online:2022-01-12 Published:2021-06-28
  • Contact: LI Zhen-Hua E-mail:lixing_19841014@126.com
  • Supported by:
    National Natural Science Foundation of China(31860420);National Natural Science Foundation of China(32060512);Natural Science Foundation of Guizhou Province (QKHPT)([2019]1069);Science and Technology Plan Project of Guizhou Province (QKHJC)([2018]5781号);High-level Talents Introduction Project of Guizhou University (GDRJHZ)([2018]37号)

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Abstract:

Temperature and light are two important environmental factors affecting seed germination, and phytochromes are the sensors of light and temperature. It is of great significance to investigate the interaction of phytochromes with light and temperature to regulate seed germination for guiding crop sowing. The aim of this study is to explore how the NtPHYB1 genotype regulates seed germination in response to change in light and temperature environments in tobacco. The average germination frequency of WT seeds was significantly higher than that of NtPHYB1-OE and NtPHYB1-RNAi seeds under 15 environments. The germination of NtPHYB1-OE seeds was inhibited by continuous light exposure, while the germination of NtPHYB1-RNAi seeds was repressed by darkness. At 15℃, the germinations of all three genotypic seeds were inhibited by the low-temperature, and the germination frequency of NtPHYB1-OE seeds was significantly lower than that of WT and NtPHYB1-RNAi seeds; while there was no significant effect in light signal. At 20℃ and 25℃, the temperature signal promoted the ability of seed germination, and the signal of light was dispensable, resulting in the highest germination frequencies for NtPHYB1-OE and WT seeds. At 30℃ and 35℃, the light signal was indispensable to maintain seed germination for all three genotypic seeds, resulting in the highest germination frequency for NtPHYB1-RNAi seeds. To sum up, there were interactions among NtPHYB1, photoperiod, and temperature in the regulation of seed germination in tobacco. The germination frequencies of tobacco seeds could be significantly improved by modifying NtPHYB1 gene or improving the cultivation environment.

Key words: seed germination, photothermal sensitivity, phytochrome, photothermal signal, natural selection, seasonal variation

Table 1

Effects of genotype, light, temperature and their interactions on seed germination in tobacco"

基因型
Genotype (G)		 光
Light treatment (L)		 温度
Temperature
(T, ℃)		 均值(L×T×G)
Mean value (L×T×G)		 标准偏差
Standard
deviation		 样本数
Sample number
NtPHYB1过表达 持续黑暗Dark 15 17.78 qrstu NOP 8.36 9
NtPHYB1-OE 20 72.00 abcdefg ABCDE 9.26 9
25 78.00 abcdef ABC 7.91 9
30 26.78 opqr LMNO 13.14 9
35 2.89 u Q 2.62 9
NtPHYB1 RNA干扰 15 31.83 nopq KLMN 4.92 6
NtPHYB1-RNAi 20 65.67 cdefghi BCDEFG 7.50 6
25 75.00 abcdef ABCD 10.12 6
30 46.00 lmn GHIJKL 8.67 6
35 5.00 tu Q 1.41 6
野生型WT 15 38.00 mno IJKLM 6.93 3
20 85.67 ab AB 2.08 3
25 80.67 abc ABC 8.14 3
30 62.00 fghij CDEFGH 3.46 3
35 5.00 tu Q 2.00 3
NtPHYB1过表达 12 h光照/12 h 黑暗 15 16.22 rstu NOP 10.97 9
NtPHYB1-OE 12 h light/12 h dark 20 48.33 klm FGHIJK 15.36 9
基因型
Genotype (G)		 光
Light treatment (L)		 温度
Temperature
(T, ℃)		 均值(L×T×G)
Mean value (L×T×G)		 标准偏差
Standard
deviation		 样本数
Sample number
25 63.33 efghi CDEFGH 10.55 9
30 74.11 abcdef ABCD 6.41 9
35 73.67 abcdef ABCD 10.32 9
NtPHYB1 RNA干扰 15 18.83 qrst MNOP 5.12 6
NtPHYB1-RNAi 20 55.00 hijkl DEFGHI 7.90 6
25 64.17 defghi CDEFGH 11.96 6
30 78.50 abcde ABC 9.67 6
35 79.33 abcde ABC 4.37 6
野生型WT 15 21.67 pqrs MNOP 4.93 3
20 69.33 cdefgh ABCDE 7.51 3
25 75.00 abcdef ABCD 2.65 3
30 79.67 abcd ABC 4.51 3
35 87.33 aA 3.06 3
NtPHYB1过表达 持续光照Light 15 8.67 stu OP 6.02 9
NtPHYB1- OE 20 38.11 mno IJKLM 18.31 9
25 53.11 jkl EFGHIJ 22.13 9
30 57.22 ghijk DEFGHI 10.93 9
35 65.00 cdefghi BCDEFG 17.68 9
NtPHYB1 RNA干扰 15 20.67 pqrs MNOP 8.98 6
NtPHYB1-RNAi 20 44.83 lmn HIJKL 7.88 6
25 68.33 cdefghi ABCDEF 8.07 6
30 70.33 bcdefgh ABCDE 11.33 6
35 66.17 cdefghi BCDEF 9.79 6
野生型WT 15 11.33 stu OP 3.51 3
20 34.00 mnop JKLMN 5.29 3
25 65.00 cdefghi BCDEFG 2.65 3
30 74.00 abcdef ABCDE 3.00 3
35 70.67 bcdefgh ABCDE 6.43 3

Table 2

Comparisons of mean difference within the genotype, light, and temperature treatment"

来源
Source		 I J 平均差
Mean difference (I-J)
基因型Genotype NtPHYB1过表达NtPHYB1-OE NtPHYB1 RNA干扰NtPHYB1-RNAi -6.30***
野生型WT -10.94***
NtPHYB1 RNA干扰NtPHYB1-RNAi NtPHYB1过表达NtPHYB1-OE 6.30***
野生型WT -4.64*
野生型WT NtPHYB1过表达NtPHYB1-OE 10.94***
NtPHYB1 RNA干扰NtPHYB1-RNAi 4.64*
光Light 持续黑暗Dark 12 h光照/12 h黑暗 12 h light/12 h dark -14.70***
持续光照Light -5.04**
来源
Source		 I J 平均差
Mean difference (I-J)
12 h光照/12 h黑暗 12 h light/12 h dark 持续黑暗Dark 14.70***
持续光照Light 9.66***
持续光照Light 持续黑暗Dark 5.04**
12 h光照/12 h黑暗 12 h light/12 h dark -9.66***
温度Temperature 15℃ 20℃ -36.31***
25℃ -48.74***
30℃ -41.00***
35℃ -30.39***
20℃ 15℃ 36.31***
25℃ -12.43***
30℃ -4.69*
35℃ 5.93**
25℃ 15℃ 48.74***
20℃ 12.43***
30℃ 7.74***
35℃ 18.35***
30℃ 15℃ 41.00***
20℃ 4.69*
25℃ -7.74***
35℃ 10.61***
35℃ 15℃ 30.39***
20℃ -5.93**
25℃ -18.35***
30℃ -10.61***

Table 3

Multi-factor variance analysis of the interactions among NtPHYB1, light, and temperature in the regulation of seed germination in tobacco"

来源
Source		 III 型平方和
III sum of squares		 自由度
DF		 均方差
MS		 F值
F-value		 显著性水平
Significant
基因型Genotype (G) 4800.759 2 2400.380 22.199*** 0.000
光处理Light (L) 7936.154 2 3968.077 36.697*** 0.000
温度处理Temperature (T) 63,436.466 4 15,859.116 146.666*** 0.000
G×L 975.230 4 243.807 2.255 0.064
G×T 1206.704 8 150.838 1.395 0.200
L×T 61,711.869 8 7713.984 71.339*** 0.000
G×L×T 3070.141 16 191.884 1.775* 0.036

Fig.1

Variation of seed germination under fixed temperature with photoperiodic change (A) and fixed photoperiodic with temperature change (B) in tobacco"

Table 4

NtPHYB1 regulates seed germination in response to signals of light and temperature"

不同基因型种子萌发差异
Germination ratio among different genotype (G)		 光和温度的环境组合
Environmental combinations of light and temperature
无显著差异
No significant difference		 NtPHYB1-OE, WT and NtPHYB1-RNAi: T15L12, T15L24, T20L24, T25L0, T25L12, T30L12, T35L0, T35L12, and T35L24.
显著差异
Significant difference		 WT>NtPHYB1-OE: T15L0, T20L12, T25L24,T30L0, T30L24; WT>NtPHYB1-RNAi: T20L0 and T30L0; NtPHYB1-RNAi>NtPHYB1-OE: T25L24.
极大值
Maximum		 NtPHYB1-OE: T20L0, T25L0, T30L12, T35L12; WT: T20L0, T25L0, T25L12, T30L12, T30L24, T35L12; NtPHYB1-RNAi: T30L12, T30L24, T35L12.
极小值
Minimum		 NtPHYB1-OE: T15L0, T15L12, T15L24, T35L0; WT: T15L24, T35L0; NtPHYB1-RNAi: T35L0

Fig. 2

Diagram of phytochrome B integrates light and temperature signal to regulate seed germination in different seasons The pink block are seeds exposed to the ground, while the grey blocks are seeds buried underground. represents quick, slow, and obstructive conversions. In winter and early spring (<15℃), the low-temperature signal sensed by phytochrome B inhibited seeds germination, and the effect of the light signal was almost ineffective. During late spring and autumn (~20℃), temperature signal initiated germination with the combination of the light. In summer and early autumn (>30℃), light is indispensable to maintain seed germination by persistently activating the PHYB."

[1] Jung J H, Domijan M, Klose C, Biswas S, Ezer D, Gao M, Khattak A K, Box M S, Charoensawan V, Cortijo S, Kumar M, Grant A, Locke J C, Schafer E, Jaeger K E, Wigge P A. Phytochromes function as thermosensors in Arabidopsis. Science, 2016, 354:886-889.
[2] Legris M, Klose C, Burgie E S, Rojas C C, Neme M, Hiltbrunner A, Wigge P A, Schafer E, Vierstra R D, Casal J J. Phytochrome B integrates light and temperature signals in Arabidopsis. Science, 2016, 354:897-900.
[3] Lee K P, Piskurewicz U, Tureckova V, Carat S, Chappuis R, Strnad M, Fankhauser C, Lopez-Molina L. Spatially and genetically distinct control of seed germination by phytochromes A and B. Gene Dev, 2012, 26:1984-1996.
[4] de Wit M, Galvao V C, Fankhauser C. Light-mediated hormonal regulation of plant growth and development. Annu Rev Plant Biol, 2016, 67:513-537.
[5] Hennig L, Poppe C, Sweere U, Martin A, Schafer E. Negative interference of endogenous phytochrome B with phytochrome A function in Arabidopsis. Plant Physiol, 2001, 125:1036-1044.
[6] Franklin K A, Praekelt U, Stoddart W M, Billingham O E, Halliday K J, Whitelam G C. Phytochromes B, D, and E act redundantly to control multiple physiological responses in Arabidopsis. Plant Physiol, 2003, 131:1340-1346.
[7] Dechaine J M, Gardner G, Weinig C. Phytochromes differentially regulate seed germination responses to light quality and temperature cues during seed maturation. Plant Cell Environ, 2009, 32:1297-1309.
[8] Strasser B, Sanchez-Lamas M, Yanovsky M J, Casal J J, Cerdan P D. Arabidopsis thaliana life without phytochromes. Proc Natl Acad Sci USA, 2010, 107:4776-4781.
[9] Heschel M S, Selby J, Butler C, Whitelam G C, Sharrock R A, Donohue K. A new role for phytochromes in temperature- dependent germination. New Phytol, 2007, 174:735-741.
[10] Heschel M S, Butler C M, Barua D, Chiang G C K, Wheeler A, Sharrock R A, Whitelam G C, Donohue K. New roles of phytochromes during seed germination. Int J Plant Sci, 2008, 169:531-540.
[11] Donohue K, Heschel M S, Chiang G C, Butler C M, Barua D. Phytochrome mediates germination responses to multiple seasonal cues. Plant Cell Environ, 2007, 30:202-212.
[12] Donohue K, Heschel M S, Butler C M, Barua D, Sharrock R A, Whitelam G C, Chiang G C. Diversification of phytochrome contributions to germination as a function of seed-maturation environment. New Phytol, 2008, 177:367-379.
[13] Borthwick H A, Hendricks S B, Parker M W, Toole E H, Toole V K. A reversible photoreaction controlling seed germination. Proc Natl Acad Sci USA, 1952, 38:662-666.
[14] Takaki M, Heeringa G H, Cone J W, Kendrick R E. Analysis of the effect of light and temperature on the fluence response curves for germination of Rumexobtusifolius. Plant Physiol, 1985, 77:731-734.
[15] Halliday K J, Davis S J. Light-sensing phytochromes feel the heat. Science, 2016, 354:832-833.
[16] Sellaro R, Smith R W, Legris M, Fleck C, Casal J J. Phytochrome B dynamics departs from photoequilibrium in the field. Plant Cell Environ, 2019, 42:606-617.
[17] Arana M V, Tognacca R S, Estravis-Barcala M, Sanchez R A, Botto J F. Physiological and molecular mechanisms underlying the integration of light and temperature cues in Arabidopsis thaliana seeds. Plant Cell Environ, 2017, 40:3113-3121.
[18] 王玉芳, 贺化祥, 张明生, 彭斯文, 徐利, 杨小蕊, 翟欣. 光照、温度和盐胁迫对红花大金元种子萌发的影响. 种子, 2009, 28(12):19-22.
Wang Y F, He H X, Zhang M S, Peng S W, Xu L, Yang X R, Zhai X. Effects of light, temperature and salt stress on seed germination of Hongda (a tobacco variety). Seed, 2009, 28(12):19-22 (in Chinese with English abstract).
[19] 王树会, 黄成江. 低温对烟草种子萌发的影响. 中国种业, 2008, (5):48-49.
Wang S H, Huang C J. Effects of low temperature on seed germination of tobacco. China Seed Ind, 2008, (5):48-49 (in Chinese).
[20] 王树会, 赵高坤, 杨志强, 张红艳. 极端高温对烟草种子萌发的影响. 中国种业, 2009, (9):52-54.
Wang S H, Zhao G K, Yang Z Q, Zhang H Y. Effects of extreme temperature on seed germination of tobacco. China Seed Ind, 2009, (9):52-54 (in Chinese).
[21] 招启柏, 宋平, 王广志, 吕冰, 曹显祖. 光、温、激素对烟草种子萌发与幼苗生长的影响. 中国烟草学报, 2001, 7(4):29-32.
Zhao Q B, Song P, Wang G Z, Lyu B, Cao X Z. Effects of light, temperature and phytohormone on seed germination and seedling growth of Nicotiana tabacum. Acta Tab Sin, 2001, 7(4):29-32 (in Chinese with English abstract).
[22] Zhao J H, Han J, Zhang J, Li Z H, Yu J, Yu S Z, Guo Y S, Fu Y F, Zhang X M. NtPHYB1K326, a homologous gene of Arabidopsis PHYB, positively regulates the content of phenolic compounds in tobacco. Plant Physiol Biochem, 2016, 109:45-53.
[23] Leubner-Metzger G, Frundt C, Vogeli-Lange R, Meins F J. Class Iβ-1,3-Glucanases in the endosperm of tobacco during germination. Plant Physiol, 1995, 109:751-759.
[24] Li Z H, Ren X L, Long M J, Kong D J, Liu Y L. Capsule colour quantification-based evaluation of seed dryness and vigour during natural and artificial drying in Nicotiana tabacum. Seed Sci Technol, 2015, 43:208-217.
[25] Finch-Savage W E, Leubner-Metzger G. Seed dormancy and the control of germination. New Phytol, 2006, 71:501-523.
[26] Shinomura T, Nagatani A, Chory J, Furuya M. The Induction of seed germination in Arabidopsis thaliana is regulated principally by Phytochrome B and secondarily by phytochrome A. Plant Physiol, 1994, 104:363-371.
[27] Shinomura T, Nagatani A, Hanzawa H, Kubota M, Watanabe M, Furuya M. Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana. Proc Natl Acad Sci USA, 1996, 93:8129-8133.
[28] Hennig L, Stoddart W M, Dieterle M, Whitelam G C, Schafe E. Phytochrome E controls light-induced germination of Arabidopsis. Plant Physiol, 2002, 128:194-200.
[29] Appenroth K J, Lenk G, Goldau L, Sharma R. Tomato seed germination: regulation of different response modes by phytochrome B2 and phytochrome A. Plant Cell Environ, 2006, 29:701-709.
[30] 李振华, 徐如宏, 任明见, 李鲁华. 光敏色素感知光温信号调控种子休眠与萌发研究进展. 植物生理学报, 2019, 55:539-546.
Li Z H, Xu R H, Ren M J, Li L H. Advances in phytochrome regulating seed dormancy and germination by sensing light and temperature signals. Plant Physiol J, 2019, 55:539-546 (in Chinese with English abstract).
[31] Alba R, Kelmenson P M, Cordonnier-Pratt M M, Pratt L H. The phytochrome gene family in tomato and the rapid differential evolution of this family in angiosperms. Mol Biol Evol, 2000, 17:362-373.
[32] Sharrock R A, Clack T, Goosey L. Differential activities of the Arabidopsis phyB/D/E phytochromes in complementing phyB mutant phenotypes. Plant Mol Biol, 2003, 52:135-142.
[33] Fragoso V, Oh Y, Kim S G, Gase K, Baldwin I T. Functional specialization of Nicotiana attenuate phytochromes in leaf development and flowering time. J Integr Plant Biol, 2017, 59:205-224.
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