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作物学报 ›› 2022, Vol. 48 ›› Issue (1): 99-107.doi: 10.3724/SP.J.1006.2022.04275

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

NtPHYB1与光温信号互作调控烟草种子萌发

李振华1,*(), 王显亚1, 刘一灵2, 赵杰宏3   

  1. 1贵州大学农学院, 贵州贵阳 550025
    2贵州大学山地植物资源保护与种质创新教育部重点实验室, 贵州贵阳 550025
    3贵州省烟草科学研究院 / 烟草行业分子遗传重点实验室, 贵州贵阳 550081
  • 收稿日期:2020-12-15 接受日期:2021-04-14 出版日期:2022-01-12 网络出版日期:2021-06-28
  • 通讯作者: 李振华
  • 基金资助:
    国家自然科学基金项目(31860420);国家自然科学基金项目(32060512);贵州省自然科学基金项目(黔科合基础)([2019]1069);贵州省科技计划项目(黔科合平台人才)([2018]5781号);贵州大学高层次人才引进项目(贵大人基合字)资助([2018]37号)

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 Published:2022-01-12 Published online:2021-06-28
  • Contact: LI Zhen-Hua
  • 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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摘要：

光照和温度是影响种子萌发的2个重要环境因子, 光敏色素是光和温度的受体, 研究光敏色素协同光温信号调控种子萌发对于指导作物播种具有重要意义。本研究旨在探索NtPHYB1基因型如何响应光温环境变化而调控烟草种子萌发。结果表明, 15种光温条件下萌发率的均值为野生型(wild type, WT)种子显著大于NtPHYB1-RNAi和NtPHYB1-OE种子。在持续光照下, NtPHYB1-OE种子萌发被抑制, 而在黑暗下NtPHYB1-RNAi种子萌发被抑制。在15℃, 3种基因型种子萌发均被抑制, NtPHYB1-OE种子萌发率最低; 在20℃和25℃, 光促进或抑制种子萌发作用不明显, 3种基因型种子萌发率均较高, NtPHYB1-OE和WT种子达到最大萌发率; 在30℃和35℃, 光对于种子维持较高萌发率是必不可少的, NtPHYB1-RNAi种子达到最大萌发率。综上所述, NtPHYB1、光周期和温度三者之间在调控烟草种子萌发时存在交互作用, 通过修饰NtPHYB1基因和改善播种环境均可显著提高烟草种子的萌发率。

关键词: 种子萌发, 光温敏感性, 光敏色素, 光温信号, 自然选择, 季节变化

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

表1

基因型、光照、温度及其互作对烟草种子萌发的影响"

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

表2

基因型、光照和温度处理均值的组内差值比较"

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***

表3

NtPHYB1、光和温度互作调控烟草种子萌发的多因素方差分析"

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

图1

在恒定光周期随着温度变化烟草种子萌发的变化(A)和在恒定温度随着光周期变化烟草种子萌发的变化(B)"

表4

NtPHYB1响应光照和温度信号互作调控种子萌发"

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

图2

光敏色素整合光温信号调控种子萌发响应季节变化 粉色模块是指暴露在地表的种子, 灰色模块是指埋在土壤中的种子。在冬季和早春(<15℃), 光敏色素感知低温信号抑制种子萌发, 而光信号的作用微乎其微。在晚春早秋(约20℃), 温度信号独立激活光敏色素促进种子萌发。在夏季和初秋(>30℃), 光是维持光敏色素活性和促进种子萌发所必需的。"

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