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作物学报 ›› 2022, Vol. 48 ›› Issue (8): 2016-2027.doi: 10.3724/SP.J.1006.2022.12041

• 耕作栽培·生理生化 • 上一篇    下一篇

褪黑素和茉莉酸甲酯基质育秧对水稻耐低温胁迫的调控作用

朱春权1,**(), 魏倩倩1,2,**(), 项兴佳2, 胡文君3, 徐青山1, 曹小闯1, 朱练峰1, 孔亚丽1, 刘佳4, 金千瑜1, 张均华1,*()   

  1. 1中国水稻研究所 / 水稻生物学国家重点实验室, 浙江杭州 310006
    2安徽大学, 安徽合肥 230039
    3浙江省农业科学院, 浙江杭州 310021
    4江西省农业科学院, 江西南昌 330200
  • 收稿日期:2021-06-16 接受日期:2021-11-29 出版日期:2022-08-12 网络出版日期:2021-12-28
  • 通讯作者: 张均华
  • 作者简介:朱春权, E-mail: zhuchunquan@caas.cn;
    魏倩倩, E-mail: 15955803543@163.com第一联系人：

    ** 同等贡献

  • 基金资助:
    国家自然科学基金项目(31901452);浙江省自然科学基金项目(LQ19C020007)

Regulation effects of seedling raising by melatonin and methyl jasmonate substrate on low temperature stress tolerance in rice

ZHU Chun-Quan1,**(), WEI Qian-Qian1,2,**(), XIANG Xing-Jia2, HU Wen-Jun3, XU Qing-Shan1, CAO Xiao-Chuang1, ZHU Lian-Feng1, KONG Ya-Li1, LIU Jia4, JIN Qian-Yu1, ZHANG Jun-Hua1,*()   

  1. 1State Key Laboratory of Rice Biology / China National Rice Research Institute, Hangzhou 310006, Zhejiang, China
    2Anhui University, Hefei 230039 Anhui, China
    3Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang, China
    4Jiangxi Academy of Agricultural Sciences, Nanchang 330200, Jiangxi, China
  • Received:2021-06-16 Accepted:2021-11-29 Published:2022-08-12 Published online:2021-12-28
  • Contact: ZHANG Jun-Hua
  • About author:First author contact:

    ** Contributed equally to this work

  • Supported by:
    National Natural Science Foundation of China(31901452);Natural Science Foundation of Zhejiang Province(LQ19C020007)

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摘要：

早稻育秧过程中易遭受低温冷害, 引起水稻减产。因此, 有必要研制耐低温的水稻育秧基质来保障早稻生产。本研究以我们实验室自己研制的发酵基质作为研究对象, 外源添加褪黑素和茉莉酸甲酯后, 分别在水稻萌发阶段和水稻生长7 d后进行为期3 d的低温处理, 然后测定水稻的萌发状况、理化性质和基因表达, 从而明确2种不同激素对早稻秧苗耐低温胁迫的调控机制。结果表明, 褪黑素和茉莉酸甲酯均显著提高水稻在低温条件下的发芽率和发芽势; 两者均能提高水稻在低温条件下的生长, 包括提高株高、根长、根条数、干物重、叶龄和养分含量。在低温条件下, 褪黑素和茉莉酸甲酯均能通过调节水稻体内抗氧化系统酶的活性和降低水稻体内过氧化氢与丙二醛含量来缓解低温胁迫导致的脂质过氧化损伤。褪黑素和茉莉酸甲酯均能提高低温条件下水稻体内的脯氨酸含量和叶绿素含量, 降低脱落酸的含量, 而茉莉酸甲酯则单独提高了水稻体内GA3的含量。褪黑素和茉莉酸甲酯对水稻耐冷基因存在不同的调控作用, 在低温条件下, 两者均显著上调OsCDPK7和OsLti6b, 显著下调OsWRKY45基因的表达, 褪黑素单独上调OsFer1基因的表达, 茉莉酸甲酯单独上调OsTrx23基因的表达。以上结果表明, 褪黑素和茉莉酸甲酯均能通过调控水稻体内抗氧化系统酶活、渗透物质含量、叶绿素含量、植物激素含量和耐冷基因表达提高水稻耐低温胁迫能力。

关键词: 基质, 水稻, 低温胁迫, 理化性质, 抗氧化酶, 基因表达

Abstract:

Early rice easily suffers from low temperature injury at seedling raising stage, resulting in yield reduction. Therefore, it is necessary to develop low temperature tolerant rice seedling substrate to ensure early rice production. The fermentation substrate made by ourselves was used in the present study. After melatonin and methyl jasmonate were added in the substrate, the rice was treated with low temperature for 3 days at seed germination stage and 7 days' growth stage. Then, to clarify the regulatory mechanism of these two phytohormones on the tolerance of early rice seedlings to low temperature stress, the germination status, physical and chemical properties, and gene expression of rice were explored. The results showed that both melatonin and methyl jasmonate could significantly improve the seed germination rate and germination potential of rice under low temperature. Both could improve the growth of rice, including plant height, root length, root number, dry matter weight, leaf age, and nutrient content. Under low temperature, melatonin and methyl jasmonate both reduced hydrogen peroxide and malondialdehyde content in rice by regulating the activities of antioxidant system enzymes, so as to alleviate the lipid peroxidation damage caused by low temperature stress. Melatonin and methyl jasmonate both increased the contents of proline and chlorophyll, and decreased the content of abscisic acid in rice under low temperature stress. Methyl jasmonate along increased GA3 content in rice under low temperature conditions. Melatonin and methyl jasmonate had different regulatory effects on cold tolerance genes expression in rice. Both of them significantly up-regulated the relative expression of OsCDPK7 and OsLti6b genes, and down-regulated the expression of OsWRKY45 gene in rice under low temperature conditions. In addition. The relative expression of OsFer1 gene was induced by melatonin alone, while the relative expression of OsTrx23 gene was up-regulated by methyl jasmonate alone in response to low temperature stress. The above results showed that both melatonin and methyl jasmonate could improve the low temperature stress tolerance of rice by regulating the enzyme activity of antioxidant system, osmotic substance content, chlorophyll content, plant hormone content and the expression of cold tolerance genes in rice.

Key words: substrate, rice, low temperature stress, physical and chemical properties, antioxidant enzymes, gene expression

表1

实验设计方案"

处理
Treatment		 温度Temperature (℃) 时间Time (h)
白天Day 晚上Night 白天Day 晚上Night
常温N 30 20 12 12
常温+褪黑素N+T 30 20 12 12
常温+茉莉酸甲酯N+M 30 20 12 12
低温L 13 7 12 12
低温+褪黑素L+T 13 7 12 12
低温+茉莉酸甲酯L+M 13 7 12 12

表2

水稻引物序列"

基因
Gene		 正向引物
Forward sequence (5'-3')		 正向引物
Reverse sequence (5'-3')
OsCOLD1 CAGGATATCAAAAGCTTGGATG GCAGCTATCTTTGCTTGACG
OsLti6b GCCTTAAATTGGAGCTCAGTC GTGCAGAAGATAAACTGGAGAA
OsWRKY45 TTCCTTGTTGATGTGTCGTCTCA CCCCCAGCTCATAATCAAGAAC
OsFer1 GAGGAGACTGTGTGAAGGGC ATATTGTTCCTTATTGGCTGCC
OsCDPK7 GAGGAAATCGCTGGGTTGA TTCTCATTGTTCTTCGTCCGA
OsTrx23 AGAACACCATCGTGAAGCAC CTATCTACAAGCTTGCCAGCAT
OsHistone GGTCAACTTGTTGATTCCCCTCT AACCGCAAAATCCAAAGAACG

表3

不同处理对水稻萌发的影响"

处理
Treatment		 发芽率
Germination rate (%)		 发芽势
Germination potential (%)
常温N 93.25±1.22 a 67.36±2.12 a
常温+褪黑素N+T 92.78±1.38 a 69.24±1.85 a
常温+茉莉酸甲酯N+M 93.74±1.67 a 69.58±1.97 a
低温L 70.08±1.21 d 33.81±1.04 c
低温+褪黑素L+T 77.41±1.42 c 44.52±1.18 b
低温+茉莉酸甲酯L+M 84.23±1.24 b 43.98±1.26 b

表4

不同温度处理后水稻生长及质量"

处理
Treatment		 株高
Plant height
(cm)		 根长
Root height
(cm)		 叶龄
Leaf age
(d)		 地上部干重
Shoot weigh
(g 100 plant-1)		 根部干重
Root weigh
(g 100 plant-1)		 茎基宽
Shoot width
(cm 5 plant-1)		 根条数
Number of roots
常温N 16.02±0.67 a 5.55±0.80 c 1.90±0.02 a 2.04±0.06 a 0.50±0.01 a 0.80±0.00 a 6.80±0.79 a
常温+褪黑素N+T 15.69±0.86 a 6.10±0.77 b 1.86±0.09 a 2.06±0.09 a 0.50±0.01 a 0.80±0.00 a 7.19±0.56 a
常温+茉莉酸甲酯N+M 15.88±0.91 a 7.49±0.85 a 1.90±0.00 a 2.07±0.10 a 0.51±0.01 a 0.80±0.00 a 6.55±0.69 b
低温L 7.68±0.56 c 4.69±0.68 d 1.36±0.15 c 1.40±0.06 c 0.34±0.01 c 0.80±0.00 a 5.27±0.70 d
低温+褪黑素L+T 9.38±0.92 b 6.34±0.99 b 1.71±0.16 b 1.60±0.04 b 0.43±0.01 b 0.80±0.01 a 6.50±0.51 b
低温+茉莉酸甲酯L+M 9.19±0.71 b 7.03±0.48 a 1.71±0.18 b 1.58±0.05 b 0.43±0.01 b 0.78±0.02 a 5.67±0.72 c

图1

不同处理后水稻表型(A)、体内叶绿素a (B)和叶绿素b含量(C) 处理同表1。不同小写字母表示差异达0.05显著水平。"

表5

不同温度处理后水稻植株养分含量"

处理
Treatment		 地上部氮含量
Shoot N content (g kg-1)		 地上部磷含量
Shoot P content (g kg-1)		 地上部钾含量
Shoot K content (g kg-1)
常温N 30.70±2.26 ab 7.24±0.09 a 50.82±1.25 b
常温+褪黑素N+T 30.41±1.86 ab 7.44±0.30 a 49.82±0.50 b
常温+茉莉酸甲酯N+M 32.09±1.08 a 7.34±0.28 a 55.33±1.42 a
低温L 27.35±0.28 c 5.29±0.12 d 41.91±1.22 d
低温+褪黑素L+T 28.12±0.31 c 6.25±0.21 b 45.24±1.38 c
低温+茉莉酸甲酯L+M 32.50±0.54 a 5.75±0.14 c 48.46±2.33 b

表6

不同温度处理后水稻体内抗氧化酶活性"

处理
Treatment		 超氧化物歧化酶
SOD
(U g-1 FW)		 过氧化物酶
POD
(U g-1 FW)		 过氧化氢酶
CAT
(nmol min-1 g-1 FW)		 谷胱甘肽转移酶
GST
(nmol min-1 g-1 FW)		 抗坏血酸过氧化物酶
APX
(μmol min-1 g-1 FW)
常温N 19.82±0.56 e 5782.16±254.16 d 66.26±2.89 d 66.26±1.29 d 0.13±0.01 d
常温+褪黑素N+T 21.40±0.33 d 6446.11±346.21 c 69.46±2.47 d 73.50±2.04 c 0.26±0.02 c
常温+茉莉酸甲酯N+M 21.37±0.49 d 6370.06±155.78 c 106.12±5.44 c 80.80±2.67 b 0.34±0.01 b
低温L 22.58±0.34 c 7122.18±264.79 b 154.32±10.10 b 79.54±5.09 b 0.23±0.02 c
低温+褪黑素L+T 23.44±0.39 b 8713.56±405.39 a 161.34±6.72 b 94.29±3.06 a 0.32±0.01 b
低温+茉莉酸甲酯L+M 24.51±0.41 a 9094.56±322.81 a 212.56±14.71 a 97.89±2.20 a 0.42±0.01 a

表7

不同温度处理后水稻体内丙二醛和过氧化氢含量"

处理
Treatment		 丙二醛含量
MDA content (nmol g-1 FW)		 过氧化氢含量
H2O2 content (μmol g-1 FW)
常温N 7.45±0.31 c 9.89±0.69 c
常温+褪黑素N+T 6.66±0.07 d 10.14±0.74 c
常温+茉莉酸甲酯N+M 6.25±0.31 d 9.92±0.08 c
低温L 22.85±2.72 a 15.84±0.85 a
低温+褪黑素L+T 15.19±0.50 b 12.26±0.56 b
低温+茉莉酸甲酯L+M 15.88±1.21 b 9.68±0.65 c

图2

不同处理后水稻体内脯氨酸含量(A)、可溶性蛋白含量(B)、脱落酸含量(C)和GA3含量(D) 处理同表1。不同小写字母表示差异达0.05显著水平。"

图3

不同处理下水稻体内基因表达 处理同表1。不同小写字母表示差异达0.05显著水平。"

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