Seed vigor testing by low temperature germination in early rice

doi:10.3785/j.issn.1008-9209.2019.02.142

Journal of Zhejiang University (Agriculture and Life Sciences)

2019, Vol. 45

Issue (6): 657-666 DOI: 10.3785/j.issn.1008-9209.2019.02.142

Crop cultivation & physiology

Seed vigor testing by low temperature germination in early rice

Dongdong CAO^1,²(

),Wei WU³,Shanyu CHEN¹,Yebo QIN⁴,Guanhai RUAN¹,Min LU⁵,Peili QIAN⁵,Yutao HUANG¹(

)

^1.Institute of Crop and Nuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
^2.Zhejiang Nongke Seed Co. , Ltd. , Hangzhou 310021, China
^3.Seed Management Station of Zhejiang Province, Hangzhou 310020, China
^4.Agricultural Technology Extension Center of Zhejiang Province, Hangzhou 310020, China
^5.Huzhou Keao Seed Co. , Ltd. , Huzhou 313000, Zhejiang, China

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Abstract

Taking the seeds of six early rice cultivars ‘Zhuliangyou 06’, ‘Zhuliangyou 101’, ‘Zhongzao 39’, ‘Jinzao 47’, ‘Zhongjiazao 17’ and ‘Yongxian 15’ as materials, the seed vigor of these early cultivars was determinated by the standard germination test, low temperature germination test (12 ℃, 15 ℃ and 18 ℃) and field emergence test. The results showed that the germination percentages at 18 ℃ after 4-5 days and 15 ℃ after 5-7 days had significantly positive relationship to the field emergence (FE). Especially, the correlation coefficient between germination percentage at 18 ℃ after 4 days and FE reached 0.932, revealing a fast and accurate method for measuring the seed vigor at a low temperature. The endogenous ABA content of seed at 18 ℃ after 4 days were negatively related to the germination percentages and FE. The real-time fluorescence quantitative polymerase chain reaction analysis showed the expression levels of ABA-synthesis related genes in high-vigor seeds were significantly lower than those in the low-vigor seeds. However, the most genes involved in ABA-catabolism and signal transduction related genes were highly expressed in the high-vigor seeds. Different responses in ABA metabolism and chilling signal transduction to low temperature stress might explain the seed vigor differences in the 12 early rice seed samples. The present study further reveals the internal mechanism of low temperature stress regulating seed vigor of early rice, which could provide a new theoretical basis for testing seed vigor of early rice with low temperature germination test.

Key words： early rice low temperature germination germination percentage seed vigor abscisic acid signal transduction

Received: 14 February 2019 Published: 20 January 2020

CLC:

S 311

Corresponding Authors: Yutao HUANG E-mail: jellycao@163.com;hytcsy@zju.edu.cn

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	Dongdong CAO
	Wei WU
	Shanyu CHEN
	Yebo QIN
	Guanhai RUAN
	Min LU
	Peili QIAN
	Yutao HUANG

Cite this article:

Dongdong CAO,Wei WU,Shanyu CHEN,Yebo QIN,Guanhai RUAN,Min LU,Peili QIAN,Yutao HUANG. Seed vigor testing by low temperature germination in early rice. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(6): 657-666.

URL:

http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2019.02.142 OR http://www.zjujournals.com/agr/Y2019/V45/I6/657

低温发芽测定早稻种子活力

以‘株两优06’‘株两优101’‘中早39’‘金早47’‘中嘉早17’和‘甬籼15’6个早稻品种种子为试验材料，采用标准发芽试验、低温发芽试验（12 ℃、15 ℃和18 ℃）和田间出苗试验等方法测定水稻种子活力。结果表明，18 ℃发芽4～5 d、15 ℃发芽5～7 d的发芽率与田间出苗率有着显著的正相关关系，其中18 ℃发芽4 d的发芽率与田间出苗率相关系数达到了0.932，能够有效地测定水稻种子活力。此外，18 ℃发芽4 d种子的内源脱落酸（abscisic acid, ABA）含量与当天的发芽率、田间出苗率呈显著的负相关关系。相比于低活力种子样品，各品种中高活力种子样品的ABA合成相关基因表达量大都显著降低，而ABA分解相关基因及ABA信号传导相关基因表达量大都显著提高。12个早稻种子样品之间ABA代谢、信号传导及其对低温胁迫响应基因表达量的差异，可能是低温胁迫下种子活力差异的重要原因。本研究结果进一步揭示了低温胁迫调控早稻种子活力的内在机制，为低温发芽试验测定早稻种子活力的方法提供了新的理论依据。

关键词： 早稻, 低温发芽, 发芽率, 种子活力, 脱落酸, 信号传导

Fig. 1 Change of daily maximum temperature and minimum temperature during the field emergence test

基因名称 Gene name	基因登录号 Gene accession No.	引物序列 Primer sequence
18S rRNA	AK059783	F: 5 $′$ -CTACGTCCCTGCCCTTTGTACA-3 $′$ R: 5 $′$ -ACACTTCACCGGACCATTCAA-3 $′$
OsNCED1	XM_015771176.2	F: 5 $′$ -TCCATGAGGCTTTCCTACATC-3 $′$ R: 5 $′$ -GCGTTCTTCTTCCTGCCATA-3 $′$
OsNCED3	XM_015776052.1	F: 5 $′$ -TCGCCATCACCGAGAACTA-3 $′$ R: 5 $′$ -TCTCCTGGAGCTTGAACACC-3 $′$
OsNCED4	XM_015790372.2	F: 5 $′$ -CCTCCTATAAATCTCCCCATGA-3 $′$ R: 5 $′$ -GGATTCGAATAATTTCGAGGAG-3 $′$
OsZEP1	XM_015835992.1	F: 5 $′$ -AATGCCTTTGATGTTGACCTG-3 $′$ R: 5 $′$ -TGGTCATTTGCCTTGTCAGA-3 $′$
OsAAO2	XM_006650643.2	F: 5 $′$ -TGATCCAGGGAGGGTTCAC-3 $′$ R: 5 $′$ -CTTCTCTTTGATGGGCTTCAG-3 $′$
OsAAO3	XM_015774952.1	F: 5 $′$ -TCAAGCGAATGGTGACACTT-3 $′$ R: 5 $′$ -AACTCCTCGACGCTCTTGG-3 $′$
OsABA8ox1	XM_015772227.2	F: 5 $′$ -GATGAAGACTTACGCGCTGA-3 $′$ R: 5 $′$ -CTCCTCGATGTAGCGCATCT-3 $′$
OsABA8ox2	XM_015784024.2	F: 5 $′$ -TTTGATCCTTCTAGGTTCGACTG-3 $′$ R: 5 $′$ -CCTGCTCCAAATACCAGCA-3 $′$
OsPYL3	XM_015771557.1	F: 5 $′$ -TCGTGGGAGGTGATCATAGG-3 $′$ R: 5 $′$ -TTCGGACAAGTGGTTTCCAT-3 $′$
OsPYL5	XM_015785221.2	F: 5 $′$ -TCCTGTTCACCTCGTGTGG-3 $′$ R: 5 $′$ -CTGACGAACGGCTTGTACCT-3 $′$
OsPYL9	XM_015769300.2	F: 5 $′$ -TCCGTCACCGAGTTCTCCT-3 $′$ R: 5 $′$ -ACTCGACGACGACGCAGTA-3 $′$
OsPP2C2	XM_015770920.2	F: 5 $′$ -GGTGGTATGATTGAAGGGTTG-3 $′$ R: 5 $′$ -CCAATCATCTTGTCCATTATTCG-3 $′$
OsPP2C3	XM_026022652.1	F: 5 $′$ -ACATCCTCGCTGATGACATAAC-3 $′$ R: 5 $′$ -TAAGAAACCACGTCCGCACT-3 $′$
OsSnRK2	XM_015783665.2	F: 5 $′$ -GGGAAAGACCATACCATATTGC-3 $′$ R: 5 $′$ -CGACGATGATCCTGTAATGGTA-3 $′$
OsDREB1c	XM_015786965.2	F: 5 $′$ -TCGACCAGCCGTCCTACTAC-3 $′$ R: 5 $′$ -CGTACATCTGAACGCTCT-3 $′$
OsDREB1f	XM_015790359.2	F: 5 $′$ -TGGAGGAGAAACCACGAGTT-3 $′$ R: 5 $′$ -CCAACATTGGGTCAATCTCA-3 $′$
OsDREB2a	XM_026022985.1	F: 5 $′$ -CGCCTATGGCTAGGATCATT-3 $′$ R: 5 $′$ -CTTGCCGCCTCATCGTAT-3 $′$

Table 1 Information of primer sequences

Table 2 Field performance of early rice seed samples

Table 3 Correlation analysis between germination energy (GE), germination percentage (GP), germination index (GI), vigor index (VI) and field emergence (FE) of early rice seed samples

Table 4 Correlation analysis between germination percentage (GP) under chilling stress and field emergence (FE) of early rice seed samples

Fig.2 Regression analysis between germination percentage and field emergence on the 4th day (A) and the 5th day (B) at 18 ℃ for early rice seed samples

Fig. 3 Abscisic acid (ABA), gibberellin (GA) contents and GA/ABA ratio of early rice seed samples after 4 days of germination at 18 ℃ABA and GA contents are calculated on the basis of fresh mass. Different lowercase letters above the bars indicate significant differences between different vigors of the same cultivar at the 0.05 probability level.

Table 5 Correlation analysis among ABA, GA contents, GA/ABA ratio and germination percentage after 4 days of germination at 18 ℃ and field emergence of early rice seed samples

Fig. 4 Expression levels of ABA metabolism-related genes in rice seed after 4 days of germination at 18 ℃Different lowercase letters above the bars indicate significant differences between different vigors of the same cultivar at the 0.05 probability level.

Fig. 5 Expression levels of ABA signal transduction-related genes in rice seed after 4 days of germination at 18 ℃Different lowercase letters above the bars indicate significant differences between different vigors of the same cultivar at the 0.05 probability level.


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