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作物学报 ›› 2019, Vol. 45 ›› Issue (10): 1595-1603.doi: 10.3724/SP.J.1006.2019.94024

• 研究简报 • 上一篇    下一篇

DNA甲基化参与调控马铃薯干旱胁迫响应

李鹏程1,2,毕真真1,2,梁文君1,2,孙超1,2,张俊莲1,白江平1,2,*()   

  1. 1甘肃省干旱生境作物学重点实验室 / 甘肃省作物遗传改良与种质创新重点实验室, 甘肃兰州 730070
    2甘肃农业大学农学院, 甘肃兰州 730070
  • 收稿日期:2019-02-16 接受日期:2019-05-12 出版日期:2019-10-12 网络出版日期:2019-09-10
  • 通讯作者: 白江平
  • 基金资助:
    本研究由国家自然科学基金项目(31660432);本研究由国家自然科学基金项目(31460369);国家现代农业产业技术体系(马铃薯)建设专项(CARS-09-P14);甘肃省马铃薯产业体系(GARS-03-P1);中国科学院“西部之光”人才培养计划(2014-01);兰州市科技发展计划项目(2015-3-62);甘肃省科技厅项目(18JR3RA170);甘肃省教育厅高校科研项目资助(2018A-040)

DNA methylation involved in regulating drought stress response of potato

LI Peng-Cheng1,2,BI Zhen-Zhen1,2,LIANG Wen-Jun1,2,SUN Chao1,2,ZHANG Jun-Lian1,BAI Jiang-Ping1,2,*()   

  1. 1Gansu Provincial Key Lab of Aridland Crop Science / Gansu Key Lab of Crop Improvement & Germplasm Enhancement, Lanzhou 730070, Gansu, China
    2College of Agronomy, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2019-02-16 Accepted:2019-05-12 Published:2019-10-12 Published online:2019-09-10
  • Contact: Jiang-Ping BAI
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31660432);This study was supported by the National Natural Science Foundation of China(31460369);the China Agriculture Research System(CARS-09-P14);the Gansu Potato Industry System (GARS-03-P1)(GARS-03-P1);the “Light of the West” Talent Training Program of the Chinese Academy of Sciences(2014-01);the Lanzhou Science and Technology Development Plan(2015-3-62);the Gansu Science and Technology Fund(18JR3RA170);the Gansu Provincial Department of Education University Research Project(2018A-040)

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摘要：

非生物胁迫下表观遗传对调控植物基因表达起重要作用, 但是有关马铃薯干旱胁迫下的表观遗传研究甚少。本研究以马铃薯品种大西洋、费乌瑞它、C119、C16和青薯9号为试验材料, 以MS培养基为对照以及分别添加200 mmol L -1甘露醇、60 μmol L -1甲基化抑制剂(5-azadC)和60 μmol L -1甲基化抑制剂+200 mmol L -1甘露醇, 处理24 d后对试管苗表型性状和生理指标进行综合分析。结果发现, 不同品种马铃薯对甘露醇和甲基化抑制剂响应程度趋势类似。在干旱和DNA甲基化抑制剂分别处理下, 马铃薯植株干鲜重、株高、叶片数和叶绿素含量均显著减少(P<0.05), SOD、POD、CAT活性和Pro、MDA含量均显著增加(P<0.05), 而分枝数、根长、平均根粗均无明显变化, 表明马铃薯不同性状指标在响应干旱胁迫和DNA去甲基化时, 受到的调控通路可能不同。进一步比较干旱胁迫和DNA去甲基化共同处理与分别处理下表型性状和生理指标的差异发现, 共同处理使马铃薯植株表型性状受到进一步抑制, 同时活化了SOD、POD、CAT, 并且使Pro、MDA含量增加, 表明马铃薯在响应干旱胁迫过程中, 部分表型的形成与DNA甲基化调控相关。这将为深入研究马铃薯干旱胁迫响应与表观遗传学之间的调控网络通路提供初步的理论基础。

关键词: 马铃薯, 干旱胁迫, DNA甲基化, 形态生理, 综合评价

Abstract:

Although epigenetics is essential for regulating gene expression in plant under abiotic stress, there are few reports regarding the epigenetics of potato under drought stress. In this study, five potato cultivars including Atlantic, Favorita, C119, C16, and Qingshu 9, were used to verify that DNA methylation is involved in potato drought stress response. The plantlets were cultured for 24 days on MS medium, MS medium supplemented with 60 μmol L -1 methylation inhibitor (5-azadC), 200 mmol L -1 mannitol, and 60 μmol L -1 methylation inhibitor combined with 200 mmol L -1 mannitol, respectively. Phenotypes, and physiological and biochemical traits were recorded and comprehensively analyzed. The response of five potato cultivars to mannitol and 5-azadC was similar. With treatments of mannitol or 5-azadC, the dry/fresh weight, shoot height, leaf number and chlorophyll content decreased significantly (P < 0.05), while the activities of SOD, POD, CAT, and contents of proline and MDA increased significantly (P < 0.05). No significant changes were observed in branch number, root length and average root diameter, indicating that potato traits might have different regulatory pathways in response to drought stress and DNA demethylation. Compared with the individual treatment, combined-treatment further inhibited the growth of plantlets, and increased activities of SOD, POD, CAT, and contents of MDA and Pro under drought stress and 5-azadC treatment, indicating that the formation of some phenotypes (not all) in response to drought was regulated through the DNA methylation. The results provide preliminary data for further study on epigenetics regulatory pathway of potato under drought stress.

Key words: potato, drought stress, DNA methylation, morphological physiology, comprehensive analysis

图1

不同处理下马铃薯试管苗生长状况 A~E分别为大西洋、费乌瑞它、C119、C16和青薯9号处理24 d后结果。T0: CK; T1: 5-azadC; T2: 甘露醇; T3: 5-azadC+甘露醇。每个处理选取3株用于照相。标尺为2 cm。"

表1

不同品种马铃薯各指标对甘露醇和5-azadC分别处理的响应程度"

指标
Index		 大西洋
Atlantic		 费乌瑞它Favorita C119 C16 青薯9号
Qingshu 9		 平均值
Mean
抗旱系数 Coefficients of resistance to drought
鲜重 FW 0.3685 0.3872 0.2977 0.2954 0.7387 0.4175
干重 DW 0.4218 0.6099 0.3273 0.3579 0.7604 0.4955
株高 SH 0.3537 0.4247 0.5353 0.5650 0.2972 0.4352
分枝数 NB 0.2500 0.5455 0.2545 0.4167 0.8889 0.4711
叶片数 NL 0.3421 0.3953 0.3978 0.4359 0.9615 0.5065
总根长 RL 0.7010 0.8807 0.5532 0.7902 0.7817 0.7414
平均根粗 ARD 0.9785 1.0468 0.9003 0.8730 0.9584 0.9514
超氧化物歧化酶 SOD 1.2969 1.2707 1.2415 1.4528 1.8126 1.4149
过氧化物酶 POD 1.2224 1.2177 1.4230 1.4113 2.2462 1.5041
过氧化氢酶 CAT 1.6252 1.7085 1.8966 1.7844 3.5571 2.1144
游离脯氨酸 Pro 1.2920 1.3428 1.6898 1.9928 2.3975 1.7430
丙二醛 MDA 2.9195 2.3183 2.8036 2.1572 1.7162 2.3830
叶绿素 Chl 0.5980 0.8280 0.5491 1.0459 0.9492 0.7940
综合响应系数CRC 0.9515 0.9982 0.9900 1.0445 1.3897 1.0748
指标
Index		 大西洋
Atlantic		 费乌瑞它Favorita C119 C16 青薯9号
Qingshu 9		 平均值
Mean
5-azadC响应系数 Coefficients of response to 5-azadC
鲜重 FW 0.7594 0.7300 0.4364 0.6180 0.3968 0.5881
干重 DW 0.5741 0.7590 0.4719 0.6882 0.4547 0.5896
株高 SH 0.5488 0.5679 0.5620 0.6179 0.6226 0.5839
分枝数 NB 0.5000 0.9091 0.5455 0.8333 0.4444 0.6465
叶片数 NL 0.8333 0.6357 0.6935 0.8462 0.8077 0.7633
总根长 RL 0.8466 0.6714 0.6726 0.8118 0.8412 0.7687
平均根粗 ARD 0.9325 1.1128 0.9718 0.9821 0.9572 0.9913
超氧化物歧化酶 SOD 1.1333 1.1594 1.1133 1.1076 1.5522 1.2131
过氧化物酶 POD 1.0898 1.0486 1.3302 1.1552 1.7649 1.2777
过氧化氢酶 CAT 1.5630 1.5851 1.5766 1.3145 2.1498 1.6378
游离脯氨酸 Pro 1.0258 1.0236 1.2417 1.7473 2.1548 1.4386
丙二醛 MDA 1.1743 1.0931 1.0387 1.1621 0.9300 1.0797
叶绿素 Chl 1.2042 1.7738 0.7249 1.3946 1.3714 1.2938
综合响应系数CRC 0.9373 1.0053 0.8753 1.0214 1.1114 0.9902

图2

不同处理下马铃薯试管苗表型和生理生化指标的变化 显著性分析使用Duncan’s法进行多重比较(P < 0.05)。"

表2

各综合指标特征值及贡献率"

主成分
Principal component		 抗旱性 Drought resistance 5-azadC响应性 Response to 5-azadC
PC1 PC2 PC3 PC1 PC2 PC3
特征值 Eigenvalue 8.595 2.460 1.719 6.750 2.757 2.456
贡献率 Contribution rate (%) 66.12 18.92 13.22 51.92 21.20 18.89
累计贡献率 Cumulative contribution rate (%) 66.12 85.04 98.26 51.92 73.12 92.01

表3

各表型因子载荷矩阵"

主成分
Principal component		 抗旱性 Drought resistance 5-azadC响应性 Response to 5-azadC
PC1 PC2 PC3 PC1 PC2 PC3
鲜重 FW 0.322 -0.105 0.214 -0.316 -0.114 0.207
干重 DW 0.294 -0.313 0.026 -0.322 0.185 0.288
株高 SH -0.205 0.377 -0.341 0.217 0.201 0.400
分枝数 NB 0.331 -0.092 -0.093 -0.294 0.279 0.210
叶片数 NL 0.332 0.086 0.145 0.159 -0.383 0.415
总根长 RL 0.148 -0.343 -0.546 0.187 -0.291 0.434
平均根粗 ARD 0.039 -0.627 0.022 -0.233 0.479 -0.023
超氧化物歧化酶 SOD 0.327 0.137 0.018 0.327 0.237 0.112
过氧化物酶 POD 0.315 0.179 0.200 0.374 0.138 -0.039
过氧化氢酶 CAT 0.322 0.095 0.221 0.312 0.221 -0.069
游离脯氨酸 Pro 0.273 0.382 -0.027 0.311 0.123 0.290
丙二醛 MDA -0.309 -0.068 0.304 -0.304 -0.319 0.198
叶绿素 Chl 0.214 0.111 -0.571 -0.123 0.364 0.409

表4

供试品种的综合性状指标、权重、μ(X)及综合评价值(D)"

品种
Variety		 CI(1) CI(2) CI(3) μ(X1) μ(X2) μ(X3) 综合评价值
Comprehensive
assessment value (D)		 综合排名
Order
抗旱性Drought resistance
大西洋 Atlantic -1.9419 -1.0192 0.9505 0.0421 0.2893 0.9236 0.2083 4
费乌瑞它 Favorita -0.4128 -2.0934 -1.0274 0.2524 0.0000 0.2496 0.2035 5
C119 -2.2478 1.3289 1.1746 0.0000 0.9216 1.0000 0.3120 3
C16 -0.4186 1.6201 -1.7600 0.2516 1.0000 0.0000 0.3619 2
青薯9号 Qingshu 9 5.0211 0.1637 0.6623 1.0000 0.6078 0.8254 0.9010 1
权重 Weight 0.6729 0.1926 0.1345
5-azadC响应性Response to 5-azadC
大西洋 Atlantic -0.8022 -2.2132 0.1744 0.2937 0.0000 0.6020 0.2893 5
费乌瑞它 Favorita -2.8667 2.1615 -0.1491 0.0000 1.0000 0.5272 0.3387 4
C119 0.3390 -0.4624 -2.4315 0.4560 0.4002 0.0000 0.3495 3
C16 -0.8330 -0.4878 1.8975 0.2893 0.3944 1.0000 0.4594 2
青薯9号 Qingshu 9 4.1629 1.0020 0.5086 1.0000 0.7350 0.6792 0.8731 1
权重 Weight 0.5643 0.2304 0.2053

表5

甘露醇和5-azadC单独与共同处理下马铃薯试管苗各指标比较分析"

指标
Index		 平均值 Mean 变幅 Range
T1/T0 T2/T0 T3/T0 T2/T0 vs. T1/T0 (%) T3/T0 vs. T1/T0 (%) T3/T0 vs. T2/T0 (%)
鲜重 FW 0.417 b 0.589 a 0.316 c -46.40 -29.19 -24.30
干重 DW 0.498 b 0.589 a 0.381 c -35.25 -15.46 -23.41
株高 SH 0.435 b 0.585 a 0.346 c -40.83 -25.54 -20.54
分枝数 NB 0.510 ab 0.697 a 0.339 b -51.37 -26.85 -33.52
叶片数 NL 0.508 b 0.764 a 0.364 c -52.41 -33.55 -28.39
总根长 RL 0.741 a 0.773 a 0.653 b -15.43 -4.05 -11.86
平均根粗 ARD 0.958 a 0.996 a 0.921 a -7.56 -3.84 -3.87
超氧化物歧化酶 SOD 1.426 b 1.221 c 1.656 a 35.63 16.79 16.13
过氧化物酶 POD 1.521 b 1.284 c 1.675 a 30.45 18.46 10.12
过氧化氢酶 CAT 2.190 a 1.691 b 2.570 a 51.98 29.51 17.35
游离脯氨酸 Pro 1.752 a 1.463 b 1.825 a 24.74 19.75 4.17
丙二醛 MDA 2.409 a 1.090 b 2.614 a 139.82 121.01 8.51
叶绿素 Chl 0.795 c 1.262 a 1.165 b -7.69 -37.00 46.54
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