水稻种子成熟后期高湿环境下种子休眠相关基因的表达

doi:10.3969/j.issn.1004-1524.2022.06.01

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (6): 1103-1113.DOI: 10.3969/j.issn.1004-1524.2022.06.01

水稻种子成熟后期高湿环境下种子休眠相关基因的表达

董袁袁¹^,²(), 徐恒², 张华², 张恒², 王伏林², 顾娜娜², 朱英²^,^*()

1.浙江师范大学化学与生命科学学院,浙江金华 321004
2.浙江省农业科学院病毒学与生物技术研究所,农产品质量安全危害因子与风险防控省部共建国家重点实验室浙江杭州 310021

收稿日期:2022-02-22 出版日期:2022-06-25 发布日期:2022-06-30
通讯作者: 朱英
作者简介:*朱英,E-mail: yzhuzaas@163.com
董袁袁(1997—),女,安徽合肥人,硕士研究生,主要从事植物抗逆性和稻米品质研究。E-mail: 572604462@qq.com
基金资助:
国家自然科学基金(31871590)

Dynamic profile of genes related to seed dormancy under high humidity condition during late stage of rice grain filling

DONG Yuanyuan¹^,²(), XU Heng², ZHANG Hua², ZHANG Heng², WANG Fulin², GU Nana², ZHU Ying²^,^*()

1. College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China
2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2022-02-22 Online:2022-06-25 Published:2022-06-30
Contact: ZHU Ying

摘要/Abstract

摘要：

为探讨水稻种子成熟后期持续阴雨天气影响种子休眠的分子机制,将成熟后期水稻放置于人工气候室（12 h光照/28 ℃,12 h黑暗/22 ℃,相对湿度为60%,光量子通量密度为600 μmol∙m^-2∙s^-1）并定时喷水（每3 h喷水1次,每桶水稻每次喷水500 mL,共处理5 d）模拟持续阴雨条件来处理开花后26、28、30、32 d的粳稻,检测其穗上发芽的情况。用同样处理条件对开花后30 d的水稻种子进行高湿处理,检测高湿处理后不同时间点的种子中ABA和GA生物合成和代谢,以及α-淀粉水解酶等相关基因的动态表达情况。结果显示,种子成熟度是受高湿诱导的穗发芽的一个重要影响因素,粳稻Dongjin开花后26 d与28 d的种子经高湿处理后少有萌发,开花后30 d及以上的种子高湿处理后大多数能萌发;高湿处理能够快速诱导脱落酸（abscisic acid,ABA）降解途径基因OsABA8ox1、OsABA8ox3和赤霉素（gibberellin,GA）合成基因OsGA3ox2和OsGA20ox1的表达,同时抑制ABA合成关键基因OsNECD1、OsNECD2和OsNECD3的表达;喷水处理72 h后,α-淀粉水解酶基因OsAmy1A、OsAmy3B、OsAmy3C和OsAmy3D的表达量快速上调,促进淀粉水解,为种子萌发提供物质和能量。该研究结果将为解析成熟期水稻在多雨高湿环境下诱发穗发芽的分子机制提供基础。

关键词: 高湿, 穗发芽, 水稻, 基因表达

Abstract:

To investigate the effect of high humidity on pre-harvest sprouting in the late stage of rice grain filling, milky stage rice plants, at 26, 28, 30, 32 DAP (day after pollination) respectively, were undergo high humidity treatment in the artificial climate chamber (28 ℃/22 ℃, 12 h light/12 h dark cycle, relative humidity were 60%, photosynthetic photon flux density were 600 μmol·m^-2·s^-1). Spikelet was sprayed periodically by water (spray 500 mL water once every 3 h per bots, 5 d in total) to mimic continuous rainy conditions, and the viviparity of mature seeds were then investigated. Then, developing seeds at DAP30 were undergo same treatment and the dynamic expression of ABA and GA synthesis and metabolism and α-starch hydrolase and other related genes were investigated at different time under high humidity treatment. The results indicated that seed maturity was an important factor affecting viviparity induced by high humidity. Few seeds were sprouted at DAP26 and DAP28 but most of seeds could be sprouted after DAP30 under high humidity treatments. The results showed that high humidity treatment could rapidly induce the expression of abscisic acid (ABA) degradation related genes, such as OsABA8ox1 and OsABA8ox3, and gibberellin (GA) synthesis genes such as OsGA3ox2 and OsGA20ox1, and simultaneously inhibited the expression of key ABA synthesis genes OsNECD1, OsNECD2 and OsNECD3. Many α-starch hydrolase genes, such as OsAmy1A, OsAmy3B, OsAmy3C and OsAmy3D were activated after 72 h water-spraying treatment and provided material and energy for seed germination. The results of this study would help us to understand the molecular mechanism of pre-harvest sprouting under rainy climate.

Key words: high humidity, pre-harvest spouting, rice, gene expression

中图分类号:

S511

董袁袁, 徐恒, 张华, 张恒, 王伏林, 顾娜娜, 朱英. 水稻种子成熟后期高湿环境下种子休眠相关基因的表达[J]. 浙江农业学报, 2022, 34(6): 1103-1113.

DONG Yuanyuan, XU Heng, ZHANG Hua, ZHANG Heng, WANG Fulin, GU Nana, ZHU Ying. Dynamic profile of genes related to seed dormancy under high humidity condition during late stage of rice grain filling[J]. Acta Agriculturae Zhejiangensis, 2022, 34(6): 1103-1113.

图/表 7

参考文献 24

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基因 Gene	上游引物 Forward primer (5'→3')	下游引物 Reverse primer (5'→3')	产物长度 Product length/bp
OsNCED1	AGCCTCGGTCTTCCAATTTT	CACCCAACACAAAAGCTACG	125
OsNCED2	TCATTCCAAAACACCTTCCA	TCCGGGGACCTCCTATGTAT	113
OsNCED3	ACGTGATCAAGAAGCCGTACCT	GCTGGTCGAGCGGGATCT	87
OsNCED4	GCCGAGACACGCATTGG	GTGAAGGTGGCGACAGCAA	74
OsABA8ox1	AAGCTGGCAAAACCAACATC	CCGTGCTAATACGGAATCCA	146
OsABA8ox2	CTACTGCTGATGGTGGCTGA	CCCATGGCCTTTGCTTTAT	115
OsABA8ox3	AGTACAGCCCATTCCCTGTG	ACGCCTAATCAAACCATTGC	136
OsAmy1A	GATACGACGTCGAACACCTC	CGGATCGGATACAGCTCGTTG	184
OsAmy2A	GCCGATCATCGCACCTCTTC	CGATCCCACATATCAGTGACG	150
OsAmy3A	GAGGGTCATCACCAAGATCG	TGTGTAGCTAGCTTGCGAGC	116
OsAmy3B	GATTGGGACACGGTATGACG	CTGCAGGAACTCTGAGACCG	110
OsAmy3C	AGACTTCCATGTCGTTGCTC	CAGGCACAAAATAGTCCTG	119
OsAmy3D	GTAGGCAGGCTCTCTAGCCT	CCAACGGTTACAAACTGCGTGA	91
OsAmy3E	GAAGGAAGGCCTCAGGGTTC	GCTCGTACACATCTCGCAGCA	153
OsGA20ox1	AATGAGCATGGTGGTGCAGCAGGAGCAG	GTTAACCACCAGGAAGAAGCCGTGCCTC	235
OsGA20ox2	TACTACAGGGAGTTCTTCGCGGACAGCA	TGTGCAGGCAGCTCTTATACCTCCCGTT	268
OsGA3ox1	GACGATTCACCTCAACATGTTCCCT	GGCTCTGCAGGATGAAGGTGAA	104
OsGA3ox2	TCTCCAAGCTCATGTGGTCCGAGGGCTA	TGGAGCACGAAGGTGAAGAAGCCCGAGT	346
OsABI3	GAAGACGGACAAGAACCTGC	ATGTTCCACACCTGAGACGT	177
OsABI4	TTCCATCACCAACCGTTC	TTGAGGAAGAGATCGAACCA	353
OsABI5	ATGGGATCTGGCATGGTCAA	CAATCGCCATCCCGTTGTAC	151
OsLEA3	GCCGTGAATGATTTCCCTTTG	CACACCCGTCAGAAATCCTCC	148
OsSLR1	GATCGTCACCGTGGTAGAGC	GAGGGAATCGAACATGGTGG	103
OsGAMYB	GAATCCACCCCTCCTGTT	GCCCCATTACTTGCTCTCC	106
OsKO2	CGTGGAGAGGCAAAGACATG	GCCAACAATGAGCTGGAACA	116
OsKS1	TCTCATTTGCACTGGGACCT	ACTGTTCAGCTTTCCCTCCA	182
OsPDS	AAACCGTTCAATGCTGGAGTTG	CAAGGTTCACAGTCCGGGATAG	210
OsZDS	CAATCATTGTACCGCGAGTCAC	TCGACAATGAGCTTCTTTCGGA	209
OsCRTISO	GCTGAAGTCCCTTGAGGAGCCTC	CCCCCGAAGTGCCTATCGCAC	240
UBQ10	TGGTCAGTAATCAGCCAGTTTGG	GCACCACAAATACTTGACGAACAG	81

基因 Gene	上游引物 Forward primer (5'→3')	下游引物 Reverse primer (5'→3')	产物长度 Product length/bp
OsNCED1	AGCCTCGGTCTTCCAATTTT	CACCCAACACAAAAGCTACG	125
OsNCED2	TCATTCCAAAACACCTTCCA	TCCGGGGACCTCCTATGTAT	113
OsNCED3	ACGTGATCAAGAAGCCGTACCT	GCTGGTCGAGCGGGATCT	87
OsNCED4	GCCGAGACACGCATTGG	GTGAAGGTGGCGACAGCAA	74
OsABA8ox1	AAGCTGGCAAAACCAACATC	CCGTGCTAATACGGAATCCA	146
OsABA8ox2	CTACTGCTGATGGTGGCTGA	CCCATGGCCTTTGCTTTAT	115
OsABA8ox3	AGTACAGCCCATTCCCTGTG	ACGCCTAATCAAACCATTGC	136
OsAmy1A	GATACGACGTCGAACACCTC	CGGATCGGATACAGCTCGTTG	184
OsAmy2A	GCCGATCATCGCACCTCTTC	CGATCCCACATATCAGTGACG	150
OsAmy3A	GAGGGTCATCACCAAGATCG	TGTGTAGCTAGCTTGCGAGC	116
OsAmy3B	GATTGGGACACGGTATGACG	CTGCAGGAACTCTGAGACCG	110
OsAmy3C	AGACTTCCATGTCGTTGCTC	CAGGCACAAAATAGTCCTG	119
OsAmy3D	GTAGGCAGGCTCTCTAGCCT	CCAACGGTTACAAACTGCGTGA	91
OsAmy3E	GAAGGAAGGCCTCAGGGTTC	GCTCGTACACATCTCGCAGCA	153
OsGA20ox1	AATGAGCATGGTGGTGCAGCAGGAGCAG	GTTAACCACCAGGAAGAAGCCGTGCCTC	235
OsGA20ox2	TACTACAGGGAGTTCTTCGCGGACAGCA	TGTGCAGGCAGCTCTTATACCTCCCGTT	268
OsGA3ox1	GACGATTCACCTCAACATGTTCCCT	GGCTCTGCAGGATGAAGGTGAA	104
OsGA3ox2	TCTCCAAGCTCATGTGGTCCGAGGGCTA	TGGAGCACGAAGGTGAAGAAGCCCGAGT	346
OsABI3	GAAGACGGACAAGAACCTGC	ATGTTCCACACCTGAGACGT	177
OsABI4	TTCCATCACCAACCGTTC	TTGAGGAAGAGATCGAACCA	353
OsABI5	ATGGGATCTGGCATGGTCAA	CAATCGCCATCCCGTTGTAC	151
OsLEA3	GCCGTGAATGATTTCCCTTTG	CACACCCGTCAGAAATCCTCC	148
OsSLR1	GATCGTCACCGTGGTAGAGC	GAGGGAATCGAACATGGTGG	103
OsGAMYB	GAATCCACCCCTCCTGTT	GCCCCATTACTTGCTCTCC	106
OsKO2	CGTGGAGAGGCAAAGACATG	GCCAACAATGAGCTGGAACA	116
OsKS1	TCTCATTTGCACTGGGACCT	ACTGTTCAGCTTTCCCTCCA	182
OsPDS	AAACCGTTCAATGCTGGAGTTG	CAAGGTTCACAGTCCGGGATAG	210
OsZDS	CAATCATTGTACCGCGAGTCAC	TCGACAATGAGCTTCTTTCGGA	209
OsCRTISO	GCTGAAGTCCCTTGAGGAGCCTC	CCCCCGAAGTGCCTATCGCAC	240
UBQ10	TGGTCAGTAATCAGCCAGTTTGG	GCACCACAAATACTTGACGAACAG	81

水稻种子成熟后期高湿环境下种子休眠相关基因的表达

Dynamic profile of genes related to seed dormancy under high humidity condition during late stage of rice grain filling

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