水杨酸引发提高低温下水稻种子萌发活力的生理与分子效应

doi:10.3864/j.issn.0578-1752.2024.07.002

Abstract

Abstract:

【Objective】The study investigated the impact of salicylic acid (SA) priming on the germination vigor and physiological response of rice seeds under low temperatures. It aimed to reveal the expression patterns of genes related to abscisic acid (ABA) and gibberellin (GA) metabolic pathways as well as cell wall relaxation genes by SA priming. This research provided a theoretical basis for the study of rice seed germination at low temperatures.【Method】Using indica three-line hybrid rice Taifengyou 208 seeds as materials, the effects of SA on seed germination vigor and physiology responses under low temperature were analyzed through seed priming treatment, and the expression patterns of genes related to ABA, GA and expansin in response to SA were analyzed by qRT-PCR.【Result】Low temperature (15 ℃) significantly delayed the germination process of rice seeds. In seeds germinated at low temperatures for one day, the endogenous SA concentration was 1.7 times higher than that at normal temperatures (28 ℃). However, for five-day-old seedlings, the SA concentration under low temperature was only 0.6% of that at normal temperatures. SA could effectively enhanced germination vigor of seeds at low temperature, with the most significant effects observed at 2 000 μmol·L^-1 SA. This concentration significantly increased the germination index, vigor index, shoot length, root length, fresh weight, and dry weight of seeds under low temperature conditions. Notably, the vigor index was three times that of non-primed seeds (CK1) and two times that of water-primed seeds (CK2). In terms of physiological indexes, SA priming increased the contents of soluble sugar, proline and active oxygen, enhanced the activities of total amylase, β-amylase, superoxide dismutase (SOD) and catalase (CAT), and decreased the content of malondialdehyde (MDA). Compared with CK1, 2 000 μmol·L^-1 SA decreased the ABA content by 79%, and increased the IAA and GA₁ contents by 32.2% and 2.66 times, respectively. In terms of gene expression, the expression levels of ABA synthesizing genes OsNCED2 and OsNCED3 were decreased by 94.26% and 90.24% compared with CK1 in seeds primed by 2 000 μmol·L^-1 SA, respectively, whereas the expression levels of ABA decomposing genes OsABA8’ox2 and OsABA8’ox3 were 5.9 and 3.9 times higher than that of CK1, respectively. Compared with CK1, SA priming significantly upregulated the expression of GA synthesizing genes OsCPS1, OsKAO and OsGA20ox1, while it significantly downregulated the expression of GA decomposing genes OsGA2ox2 and OsGA2ox6. In several candidate genes encoding cell wall relaxation protein, e.t. expansin, all but OsEXPB11 were significantly upregulated to some extent by priming. Compared with CK1, 2 000 μmol·L^-1 SA increased the expression levels of OsEXPA2, OsEXPB4 and OsEXPB6 to 12.2, 5.9 and 6.1 times, respectively.【Conclusion】SA priming can significantly alleviate the impact of low temperatures on rice seed germination and seedling growth, which is likely due to SA enhancing the activity of antioxidant enzymes such as SOD and CAT, reducing the production of MDA, and increasing the content of soluble sugars and proline, thereby strengthening the tolerance of seeds and seedlings to low temperatures. On the other hand, SA priming decreases endogenous ABA content, increases GA₁ content, enhances the activities of total amylase and β-amylase, and promotes the expression of genes related to cell wall relaxation, thus facilitating seed germination and seedling growth at low temperature.

Key words: rice, low temperature stress, seed priming, salicylic acid (SA), physiological index, gene expression

CHEN BingXian, ZHANG Qi, DAI ZhangYan, ZHOU Xu, LIU Jun. Physiological and Molecular Effects of Salicylic Acid on Rice Seed Germination at Low Temperature[J].Scientia Agricultura Sinica, 2024, 57(7): 1220-1236.

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References 52

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基因名称 Gene name	基因登录号 Gene ID	正向引物 Forward primer (5′-3′)	反向引物 Reverse primer (5′-3′)
OsNCED1	LOC_Os02g47510	GAACTTCGACTTCCCCGTGA	ACGTAGCACACCAAGTACCC
OsNCED2	LOC_Os12g24800	GAGCCTTGAGTTCGGTGTCA	CAGCAAAGCACCCTAGACCA
OsNCED3	LOC_Os03g44380	ATATGGCGACGATCACGACG	CGCGGAGAATCTCACCGAAT
OsNCED4	LOC_Os07g05940	TCGGGAGGTACGACTTCCAT	TTGAGGTACGGCTTGGACAC
OsNCED5	LOC_Os12g42280	CGAGCTCACCAAGTTCGAGT	TTGATGAAGGTGCCGTGGAA
OsABA8’ox1	LOC_Os02g47470	AAAACCAACATCAACGGCGG	GATTGCCAACCGTGGTCCTA
OsABA8’ox2	LOC_Os08g36860	CGTGTGTGTGGATGCAATGG	AGTGCTACACTAGGCACACC
OsABA8’ox3	LOC_Os09g28390	CTGGTCACTGGCTACAGGTG	TGCTACGCCATTGTCGTCAT
OsCPS1	LOC_Os02g17780	TCAAGAGACACCGCCAGTTC	ACAGTGCATGACCCTGGATG
OsKAO	LOC_Os06g 02019	CTCCTTCGTGTCCTTCCGTC	ACCACAGCTGAACCTTCCAC
OsGA20ox1	LOC_Os03g63970	TTCTTCCTCTGCCCGGAGAT	CATGTCGGCCCTGTAGTGG
OsGA3ox2	LOC_ Os01g08220	CTTCTGTGACGTGATGGAGGAG	CTCAAGAACAACCTCAGCAACTC
OsGA2ox1	LOC_Os05g06670	ACCCGCAGATCCTTAGCTTG	TGAACCCACATCTCCTTGCC
OsGA2ox2	LOC_Os01g22910	TGTTTGGTTGAGGGGTGAGT	CATTTTCCCCGATCAGTTGGT
OsGA2ox3	LOC_Os01g55240	ACTCGTTGCAGGTTCTGACC	GTGGCAATGGTGCAATCCTC
OsGA2ox6	LOC_Os04g44150	GGCTATCGGTGGCCTACTTC	TTGTCCTGACGTCTTCCTGC
OsEXPA2	LOC_Os01g60770	GCGGCCAGTTCTGATCGAGTA	GCAGCCTCAGAATAGCCAAAGC
OsEXPA4	LOC_Os05g39990	CCGTCTCCGACACCCACATAT	TGGACGAAGTCCAGAGAAGGAA
OsEXPB4	LOC_Os10g40730	CCCAACACATTCTACCGCTCCT	ACAGACCGACCACACAATCCC
OsEXPB6	LOC_Os10g40700	AATTTGCGTGGGATTGAGGTGT	TGGGTAGTACAGTGACAGTGGG
OsEXPB11	LOC_ Os02g44108	TGCAGTGCAGAGTTGCGGTAA	CAGAGACCGTGGAGGGAAGAAC
OsEXPLA1	LOC_Os03g04020	ACACGCACGAGTGGAAGTAGAA	TGCCGAGGGATTAGGAGGACT
OsGAPDH1	LOC_Os02g38920	GCAATCAAGGAGGAGGCTGA	ACGTGTCGCTCAAAGCAATG

引发方式 Priming method	发芽势 Germination potential (%)	发芽率 Germination rate (%)	发芽指数 Germination index	活力指数 Vigor index	芽长 Seedling length (cm)	根长 Root length (cm)	鲜重 Fresh weight (g)	干重 Dry weigh (g)
CK1	43.75±3.11e	84.75±3.30d	15.26±2.84d	19.70±2.84e	1.13±0.29d	1.20±0.21d	0.44±0.01d	0.40±0.01bc
CK2	52.25±3.56d	84.25±3.20d	16.81±2.21cd	30.25±3.21d	1.21±0.07d	1.77±0.12c	0.47±0.01c	0.38±0.02c
50 μmol·L^-1 SA	55.00±3.92d	85.50±1.29d	16.17±1.90d	31.60±3.90d	1.47±0.21c	1.83±0.22c	0.43±0.02d	0.39±0.01c
500 μmol·L^-1 SA	90.75±4.27a	90.75±2.75c	17.32±1.98bc	36.37±4.98c	1.49±0.18c	2.10±0.22b	0.54±0.02b	0.42±0.02b
2000 μmol·L^-1 SA	83.25±3.30b	96.50±1.73a	21.59±2.67a	60.20±4.67a	1.65±0.05a	2.76±0.14a	0.63±0.02a	0.55±0.04a
5000 μmol·L^-1 SA	63.50±3.30c	92.50±1.29b	18.36±1.85b	48.40±3.05b	1.57±0.19b	2.69±0.24a	0.61±0.03a	0.54±0.02a

Physiological and Molecular Effects of Salicylic Acid on Rice Seed Germination at Low Temperature

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