Germination characteristics and antioxidant responses of alfalfa seeds to aging treatment

doi:10.11686/cyxb2024506

Abstract

Abstract:

The aims of this study were to investigate the germination characteristics of aged alfalfa （Medicago sativa） seeds， the physiological changes in their antioxidant system during the early stage of germination， and the responses of antioxidant-related genes to seed aging. We observed seed germination characteristics and determined antioxidant enzyme activities and antioxidant contents during the early stage of germination following an aging treatment， and analyzed the transcript profiles of genes related to the antioxidant system. The results show that an aging treatment not only inhibited the germination of alfalfa seeds， but also significantly affected seedling growth after germination， leading to smaller seedlings. The aging treatment also severely impacted the metabolism of reactive oxygen species， resulting in increased H₂O₂ levels during early germination of aged seeds， a significant decrease in the activities of the antioxidant enzymes peroxidase and glutathione reductase， and a marked reduction in the contents of the antioxidants ascorbic acid and glutathione. The decrease in antioxidant enzyme activity and antioxidant content reduced the seeds’ antioxidant capacity. Additionally， the aging treatment led to significantly lower transcript levels of MsCAT1， MsPOD12， MsDHAR， MsGR1， MsFe-SOD， and MsMn-SOD，which encode important enzymes in the antioxidant system. On the basis of analyses of antioxidant physiology and gene transcript profiles， several key candidate genes in the antioxidant system were identified. These genes will be important targets for further research on the molecular regulation of seed aging. The results of this study lay the foundation for further studies on the precise pathways through which these candidate genes regulate seed vigor.

Key words: alfalfa, germination characteristics, early germination stage, antioxidant enzyme, antioxidant, molecular response

Shou-jiang SUN, Hao-zhen LIU, Shu-han XU, Jing-hong ZHANG, Shu-xia LI, Jin-qing ZHANG, Xue-qin GAO, Bing-zhe FU. Germination characteristics and antioxidant responses of alfalfa seeds to aging treatment[J]. Acta Prataculturae Sinica, 2025, 34(11): 195-204.

Figures/Tables 6

References 32

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编号 No.	基因名称 Gene name	引物序列 Primer sequence （5′-3′）	产物大小 Product size （bp）	溶解温度 Melting temperature （℃）
1	MsCAT1	F： ACAGGGATGAGGAGGTGAACTAC R： CGCTGGATGAAACGGTCTTGC	130	63.6 63.3
2	MsPOD12	F： CCCAAGGCAGAGTCCATCGTG R： CCGTCCAGGAGCACTGAAGC	102	60.0 59.8
3	MsAPX	F： AAGGACTATGCTGAATCACACAAG R： CAACTGCTACGCCGACTGC	111	56.9 58.5
4	MsDHAR	F： AAGACCAAGCCAGCCAAGGAG R： AAGGACAGGACAGAACACAGACG	96	60.2 60.9
5	MsCu/Zn-SOD	F： ACCGCCGTGACAGGAAGC R： GCAACACCATCCGCTCCAG	85	60.7 60.5
6	MsFe-SOD	F： AGAAGCACCACGCCACCTAC R： CGCCGCCGTTGAACTTGATG	112	59.1 59.8
7	MsMn-SOD	F： GCTGATGTGCTTGAATCCTTGAAC R： GTGCTTGTGCTGGTGAACTCC	147	59.7 59.0
8	MsGR1	F： AGAACAGCATATCCAAACGACAG R： CAGCCTTGAGTGCGACAGC	133	60.0 59.7
9	MsAOX1c	F： GCTGGAGGAGGCGGAGAAC R： TCGGTGTAGGAGTGGATGGC	96	58.4 56.5
10	MsNAC67	F： TGCCAAGCCTGCCCTGATATG R： ATTATTGTTGCCCACCTCGTTTCC	142	58.8 59.7
11	MsNAC74	F： CCGCCTTGCCGACACCTC R： TCTTCTCCCATTCGTTCTTCTTGC	101	58.6 59.9
12	MsNAC83	F： CGGAAGGCTGGGTGCTCTG R： TCGTGGACGGGATCTTCTTGG	124	60.3 60.1
13	MsACTIN	F： CAAAAGATGGCAGATGCTGAGGAT R： CATGACACCAGTATGACGAGGTCG	88	59.4 59.5

编号 No.	基因名称 Gene name	引物序列 Primer sequence （5′-3′）	产物大小 Product size （bp）	溶解温度 Melting temperature （℃）
1	MsCAT1	F： ACAGGGATGAGGAGGTGAACTAC R： CGCTGGATGAAACGGTCTTGC	130	63.6 63.3
2	MsPOD12	F： CCCAAGGCAGAGTCCATCGTG R： CCGTCCAGGAGCACTGAAGC	102	60.0 59.8
3	MsAPX	F： AAGGACTATGCTGAATCACACAAG R： CAACTGCTACGCCGACTGC	111	56.9 58.5
4	MsDHAR	F： AAGACCAAGCCAGCCAAGGAG R： AAGGACAGGACAGAACACAGACG	96	60.2 60.9
5	MsCu/Zn-SOD	F： ACCGCCGTGACAGGAAGC R： GCAACACCATCCGCTCCAG	85	60.7 60.5
6	MsFe-SOD	F： AGAAGCACCACGCCACCTAC R： CGCCGCCGTTGAACTTGATG	112	59.1 59.8
7	MsMn-SOD	F： GCTGATGTGCTTGAATCCTTGAAC R： GTGCTTGTGCTGGTGAACTCC	147	59.7 59.0
8	MsGR1	F： AGAACAGCATATCCAAACGACAG R： CAGCCTTGAGTGCGACAGC	133	60.0 59.7
9	MsAOX1c	F： GCTGGAGGAGGCGGAGAAC R： TCGGTGTAGGAGTGGATGGC	96	58.4 56.5
10	MsNAC67	F： TGCCAAGCCTGCCCTGATATG R： ATTATTGTTGCCCACCTCGTTTCC	142	58.8 59.7
11	MsNAC74	F： CCGCCTTGCCGACACCTC R： TCTTCTCCCATTCGTTCTTCTTGC	101	58.6 59.9
12	MsNAC83	F： CGGAAGGCTGGGTGCTCTG R： TCGTGGACGGGATCTTCTTGG	124	60.3 60.1
13	MsACTIN	F： CAAAAGATGGCAGATGCTGAGGAT R： CATGACACCAGTATGACGAGGTCG	88	59.4 59.5