人工老化的甜玉米种子活力、生理特性和遗传多样性的变化

祝煜中; 夏黎明; 竺思仪; 刘继辉; 杨瑞春; 王青峰; 李小琴; 冯发强

doi:10.7671/j.issn.1001-411X.2018.01.005

人工老化的甜玉米种子活力、生理特性和遗传多样性的变化

华南农业大学农学院/广东省植物分子育种重点实验室，广东广州 510642

基金项目: 广东省科技计划项目(2015A020209113)；广州市科技计划项目(201607010248)

详细信息

作者简介:
祝煜中(1993—)，男，硕士研究生，E-mail: yuzhongzhu@foxmail.com

夏黎明(1993—)，男，硕士研究生，E-mail: xialiming1234567@sina.com；†对本文贡献相同

通讯作者:
冯发强(1976—)，男，助理研究员，博士，E-mail: fengfq@scau.edu.cn

†对本文贡献相同

中图分类号: S513
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出版历程
- 收稿日期: 2017-06-20
- 网络出版日期: 2023-05-17
- 刊出日期: 2018-01-09

Changes of vigor, physiological characteristics and genetic diversities of artificially aged sweet corn seeds

College of Agriculture, South China Agricultural University/Guangdong Provincial Key Laboratory of Plant Molecular Breeding, Guangzhou 510642, China

摘要

摘要:
目的
研究人工老化处理对甜玉米 Zea mays L. saccharata Sturt. 种子活力、生理特性和遗传多样性的影响，为甜玉米种子贮藏及大田生产提供参考。

方法
以甜玉米品种‘农甜88’和‘农甜99’为试验材料，采用人工老化的方法处理玉米种子，沙床发芽法检测种子的发芽势、发芽率、发芽指数和活力指数。测定了种子的脱氢酶(Dehydrogenase，DHA)活性、过氧化物酶(Peroxidase，POD)活性、相对电导率(Relative electrical conductivity, REC)和丙二醛(Malondialdehyde，MDA)含量，采用SRAP标记检测不同处理时间种子的遗传多样性，分析种子生理指标和遗传多样性与种子活力的关系。

结果
随老化时间的增加，2个品种种子的发芽势、发芽率、发芽指数、活力指数、DHA活性和POD活性逐渐降低，而REC和MDA含量则逐渐升高。老化处理6 d后，‘农甜88’和‘农甜99’种子的发芽势、发芽率、发芽指数、活力指数、DHA活性、POD活性分别比对照下降18.32%和43.15%、10.73%和15.38%、19.43%和23.60%、41.85%和45.70%、69.78%和71.74%、52.02%和71.60%；浸泡24 h的REC分别比对照提高了33.68%和27.68%，MDA含量分别比对照提高了41.50%和47.08%。老化处理使种子的遗传多样性下降，老化处理6 d后，‘农甜88’和‘农甜99’等位基因数、有效等位基因数、基因多样性分别比对照下降29.35%和23.58%、25.00%和20.00%、96.99%和83.35%。相关性分析发现， 2个品种老化种子浸出液的REC和MDA含量均与种子活力各性状呈显著或极显著负相关，DHA和POD活性均与种子活力各性状呈显著或极显著正相关。

结论
随老化时间的增加，2个甜玉米品种种子的活力以及DHA、POD活性逐渐降低、REC和MDA含量逐渐升高；种子老化处理降低了种子遗传物质的多样性；‘农甜88’种子耐老化能力强于‘农甜99’。
- 甜玉米 /
- 人工老化 /
- 种子活力 /
- 生理特性 /
- 遗传多样性
Abstract:
Objective
To identify the effects of artificial aging on vigor, physiological characteristics and genetic diversities of sweet corn (Zea mays L. saccharata Sturt.) seeds, and provide a reference for sweet corn seed storage and field production.

Method
‘Nongtian88’ and ‘Nongtian99’ were selected as experimental materials and their seeds were treated using the method of artificial aging. The germination potential, germination rate, germination index and vigor index were measured using the method of germination in sand bed. The dehydrogenase (DHA) activity, peroxidase (POD) activity, relative electrical conductivity (REC) and malondialdehyde (MDA) content of artificially aged seeds were measured. The genetic diversity of artificially aged seeds was detected using SRAP, and the relationships between physiological characteristics, genetic diversities and seed vigor were analyzed.

Result
With the increase of aging time, seed germination potentials, germination rates, germination indexes, vigor indexes, DHA activities and POD activities of two cultivars decreased gradually, while the RECs and MDA contents increased gradually. Compared to the control (CK), the germination potentials, germination rates, germination indexes, vigor indexes, DHA activities and POD activities of ‘Nongtian88’ and ‘Nongtian99’ seeds decreased by 18.32% and 43.15%, 10.73% and 15.38%, 19.43% and 23.60%, 41.85% and 45.70%, 69.78% and 71.74%, 52.02% and 71.60% respectively after aging treatment for six days. The RECs of ‘Nongtian88’ and ‘Nongtian99’ seeds after soaking for 24 hours increased by 33.68% and 27.68% respectively than that of CK, and MDA contents increased by 41.50% and 47.08% respectively than that of CK. Compared to CK, the seed genetic diversities decreased after artificial aging treatment, the allele numbers, effective allele numbers and gene diversities of ‘Nongtian88’ and ‘Nongtian99’ seeds decreased by 29.35% and 23.58%, 25.00% and 20.00%, 96.99% and 83.35% respectively after aging treatment for six days. The correlation analysis showed that the RECs and MDA contents of aged seeds of two cultivars were significantly or highly significantly negatively correlated with vigor indexes, while DHA and POD activities were significantly or highly significantly positively correlated with seed vigor indexes.

Conclusion
With the increase of artificially aging time, the seed vigor indexes, DHA and POD activities of two sweet corn cultivars decrease gradually, while the RECs and MDA contents increase gradually. Artificial aging results in the decrease of seed genetic diversity. ‘Nongtian88’ seed has stronger aging resistant ability than ‘Nongtian99’.
- sweet corn /
- artificial aging /
- seed vigor /
- physiological characteristic /
- genetic diversity

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图 1 甜玉米种子老化后的相对电导率(REC)

Figure 1. Relative electrical conductivity rates of sweet corn seeds after different aging time

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图 2 甜玉米种子老化后酶活性和MDA含量

Figure 2. Enzyme activities and MDA content of sweet corn seeds after different aging time

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表 1 多样性分析采用的SRAP引物名称及序列

Table 1 The SRAP primers and sequences for genetic diversity analyses

引物名称	正向引物序列	引物名称	反向引物序列
M1	TGAGTCCAAACCGGATA	E1	GACTGCGTACGAATTAAT
M2	TGAGTCCAAACCGGAGC	E2	GACTGCGTACGAATTTGC
M3	TGAGTCCAAACCGGAAT	E3	GACTGCGTACGAATTGAC
M4	TGAGTCCAAACCGGACC	E4	GACTGCGTACGAATTTGA
M5	TGAGTCCAAACCGGAAG	E5	GACTGCGTACGAATTAAC

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表 2 ‘农甜88’和‘农甜99’经不同老化处理时间后种子活力指标比较¹⁾

Table 2 Comparisons of vigor indexes of ‘Nongtian88’ and ‘Nongtian99’ seeds after different aging time

品种	t_老化/d	发芽势/%	发芽率/%	发芽指数	活力指数
农甜88	0(对照)	94.67±0.04a	99.33±0.03a	4.58±0.12a	64.78±5.31a
	1	91.33±0.02ab	95.33±0.03ab	4.42±0.16ab	61.50±5.32a
	2	85.33±0.03bcd	92.00±0.02abc	4.08±0.09bc	49.05±1.59b
	3	87.00±0.01abc	92.00±0.00abc	4.07±0.05bc	48.73±0.32b
	4	84.67±0.01bcd	91.33±0.01abc	4.05±0.04bc	44.48±0.76bc
	5	82.00±0.02cd	86.00±0.02c	3.77±0.09c	43.63±1.80bc
	6	77.33±0.02d	88.67±0.02bc	3.69±0.11c	37.67±0.83c
农甜99	0(对照)	63.33±0.02a	95.33±0.01a	3.39±0.02a	59.28±2.39a
	1	59.33±0.04a	93.33±0.02a	3.29±0.06a	53.58±2.55ab
	2	56.67±0.01a	83.33±0.01b	3.05±0.05ab	45.88±3.74abc
	3	52.67±0.09a	83.33±0.05b	2.90±0.22bc	44.92±8.13bc
	4	52.67±0.01a	82.00±0.04b	2.89±0.12bc	40.39±4.48bc
	5	46.67±0.07ab	82.67±0.02b	2.75±0.12bc	39.33±2.24c
	6	36.00±0.03b	80.67±0.02b	2.59±0.04c	32.19±0.95c
1) 同一品种、同列数据后凡是有一个相同小写字母者，表示不同处理间差异不显著(P>0.05，Duncan’s 法)

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表 3 老化处理后甜玉米种子发芽指标与生理指标的相关性¹⁾

Table 3 Correlations between germination indexes and physiological indexes of sweet corn seeds after aging treatment

发芽指标	品种	不同浸泡时间的 REC				DHA活性	POD活性	MDA含量
发芽指标	品种	4 h	8 h	12 h	24 h	DHA活性	POD活性	MDA含量
发芽势	农甜88	–0.990 11^**	–0.993 55^**	–0.992 41^**	–0.995 62^**	0.951 64^**	0.956 29^**	–0.937 54^**
发芽势	农甜99	–0.931 71	–0.923 28	–0.918 10	–0.917 32	0.899 76^**	0.891 85^**	–0.987 84^**
发芽率	农甜88	–0.993 74^**	–0.992 91^**	–0.995 00^**	–0.992 03^**	0.938 93^**	0.927 39^**	–0.877 18^**
发芽率	农甜99	–0.933 01	–0.943 27	–0.947 99	–0.946 30	0.898 27^**	0.865 46^*	–0.811 89^*
发芽指数	农甜88	–0.993 04^**	–0.996 02^**	–0.995 56^**	–0.997 84^**	0.955 65^**	0.963 71^**	–0.948 29^**
发芽指数	农甜99	–0.996 02^**	–0.993 06^**	–0.991 23^**	–0.991 78^**	0.956 06^**	0.929 07^**	–0.963 55^**
活力指数	农甜88	–0.998 38^**	–0.996 20^**	–0.990 86^**	–0.992 77^**	0.952 10^**	0.922 16^**	–0.911 62^**
活力指数	农甜99	–0.999 88^**	–0.999 46^**	–0.999 02^**	–0.999 61^**	0.970 92^**	0.955 26^**	–0.960 40^**
1) “” 和 “*” 分别表示相关性达到 0.05 和 0.01 的显著水平

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表 4 不同老化时间甜玉米种子的遗传多样性分析

Table 4 The genetic diversity analyses of sweet corn seeds after different aging time

品种	t_老化/d	等位基因数	有效等位基因数	基因多样性
农甜88	1	1.434 2	1.347 4	0.193 0
	2	1.421 1	1.336 8	0.187 1
	3	1.118 4	1.094 7	0.052 6
	4	1.065 8	1.052 6	0.029 2
	5	1.039 5	1.031 6	0.017 5
	6	1.013 2	1.010 5	0.005 8
农甜99	1	1.394 7	1.315 8	0.175 4
	2	1.289 5	1.231 6	0.128 7
	3	1.236 8	1.189 5	0.105 3
	4	1.223 7	1.178 9	0.099 4
	5	1.157 9	1.126 3	0.070 2
	6	1.065 8	1.052 6	0.029 2

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