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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (8): 2016-2027.doi: 10.3724/SP.J.1006.2022.12041

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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 Online:2022-08-12 Published:2021-12-28
  • Contact: ZHANG Jun-Hua E-mail:zhuchunquan@caas.cn;15955803543@163.com;zhangjunhua@caas.cn
  • 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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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

Table 1

Scheme of experimental designs"

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

Table 2

Sequence of primers in rice"

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

Table 3

Effects of different treatments on rice germination"

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

Table 4

Growth and weight parameters of rice under different temperature treatments"

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

Fig. 1

Rice phenotypic (A), chlorophyll a content (B), and chlorophyll b content (C) under different treatments Treatments are the same as those given in Table 1. Columns with different lowercase letters are significantly different at the 0.05 probability level."

Table 5

Nutrient contents in rice under different temperature treatments"

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

Table 6

Activity of anti-oxidase in rice under different temperature treatments"

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

Table 7

Contents of MDA and H2O2 in rice under different temperature treatments"

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

Fig. 2

Proline content (A), soluble protein content (B), abscisic acid content (C), and GA3 content (D) in rice under different treatments Treatments are the same as those given in Table 1. Columns with different lowercase letters are significantly different at the 0.05 probability level."

Fig. 3

Relative expression patterns of relevant genes in rice under different treatments Treatments are the same as those given in Table 1. Columns with different lowercase letters are significantly different at the 0.05 probability level."

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