不同水稻种质在不同生育期耐盐鉴定的差异

doi:10.11983/CBB16192

植物学报 ›› 2017, Vol. 52 ›› Issue (1): 77-88.DOI: 10.11983/CBB16192

所属专题：水稻生物学专辑 (2017年52卷1期)

不同水稻种质在不同生育期耐盐鉴定的差异

吴家富, 杨博文, 向珣朝^*(), 许亮, 颜李梅

西南科技大学植物分子遗传育种实验室, 绵阳 621010

收稿日期:2016-10-07 接受日期:2016-12-13 出版日期:2017-01-01 发布日期:2017-01-23
通讯作者: 向珣朝
作者简介:
# 共同第一作者
基金资助:
西南科技大学重点科研平台专职科研创新团队建设基金(No.14tdgc07)、四川省教育厅重点项目(No.17ZA0272)和西南科技大学实验室开放基金(No.15xnkf5)

Identification of Salt Tolerance in Different Rice Germplasm at Different Growth Stages

Jiafu Wu^†, Bowen Yang^†, Xunchao Xiang^*, Liang Xu, Limei Yan

Laboratory of Plant Molecular Genetics and Breeding, Southwest University of Science and Technology, Mianyang 621010, China

Received:2016-10-07 Accepted:2016-12-13 Online:2017-01-01 Published:2017-01-23
Contact: Xiang Xunchao
About author:
# Co-first authors

摘要/Abstract

摘要： 以21份水稻(Oryza sativa)种质为材料, 用1.5%NaCl处理种子8天后测定发芽率。在苗期用不同浓度NaCl水培处理10天, 测定叶片死亡率等指标和高亲和性K⁺转运基因(HKT)家族变异。在成株期选3份种质, 用不同浓度NaCl盆栽处理, 在开花期和籽粒蜡熟期测定植株可溶性糖和生物量等指标, 以明确各种质不同生育期的耐盐差异和关键指标。结果表明, 在NaCl胁迫下, 种子发芽率受到显著影响。苗期盐胁迫后, 各种质的平均叶片死亡率变幅最大。在被鉴定的8个耐盐种质中, HKT家族的7个基因除OsHKT2;4外均存在。在≤1 g·kg^-1盐胁迫下植株可溶性糖含量表现出刺激增长效应。CG15R单株生物量与盐浓度呈正相关, 且随盐浓度的增加而缓慢增长。在≤1 g·kg^-1时, 中花9号的生物量随盐浓度的增加而增加。水稻耐盐性具有明显的阶段发育特异性, 且不同发育阶段的耐盐性之间无相关性。叶片死亡率与蜡熟期生物量可分别作为苗期和成株期耐盐鉴定的关键指标。CG15R可作为高耐盐种质进行深入分析和利用。

Abstract: We used 21 rice (Oryza sativa) germplasm as experimental materials. Dry seeds were treated for 8 d in 1.5% NaCl solution and germination rate was measured. The seedlings with 2 green leaves were cultured by hydroponics in Petri dishes with different NaCl concentrations. After 10 days, relative shoot height, relative root length, leaf mortality, water content and the expression of 7 of 9 high-affinity K⁺ transporter (HKT) gene families were measured. In the adult period (30-d seedling to maturation), 3 representative germplasm were treated at different NaCl concentrations in a pot experiment. We investigated the effects of salt stress at flowering or dough stage of rice, for example, soluble sugar content. The results showed that germination rates of seed were affected significantly, with a wide range of variation, with 1.5% NaCl stress concentration. The variation in leaf mortality was the largest among the measured indicators at seedling stage. Besides OsHKT2;4, 6 members of HKT gene family were detected in the 8 germplasm genome. Soluble sugar content of germplasm showed a growth-stimulation effect with salt content stress ≤1 g∙kg^-1. The biomass of CG15R was positively correlated with salt content and the biomass increased with the addition of salt. Biomass of Zhonghua9 increased with the addition of salt with salt content ≤1 g∙kg^-1. Salt tolerance of rice shows the characteristic of stage development, and salt tolerance at different developmental stages is not correlated. The leaf mortality and biomass of dough stage could be the key indicators of salt tolerance for seedling and adult rice. CG15R represents a germplasm with high salt tolerance and should be further analyzed.

吴家富, 杨博文, 向珣朝, 许亮, 颜李梅. 不同水稻种质在不同生育期耐盐鉴定的差异. 植物学报, 2017, 52(1): 77-88.

Jiafu Wu, Bowen Yang, Xunchao Xiang, Liang Xu, Limei Yan. Identification of Salt Tolerance in Different Rice Germplasm at Different Growth Stages. Chinese Bulletin of Botany, 2017, 52(1): 77-88.

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图/表 10

参考文献 34

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No.	Germplasm	Subspecies	No.	Germplasm	Subspecies
1	C190	Indica	12	CG131R	Indica
2	CG133R	Indica	13	CG158R	Indica
3	HD63	Indica	14	CG111R	Indica
4	CG15R	Indica	15	CG114R	Indica
5	Javanica 22	Japonica	16	CG173R	Indica
6	Shuhui527	Indica	17	CG132R	Indica
7	3301R	Indica	18	CG151R	Indica
8	Lehui188	Indica	19	CG159R	Indica
9	Zhonghua9	Japonica	20	CG240R	Indica
10	Jing925	Japonica	21	BR207-2	Indica
11	BR207-1	Indica

No.	Germplasm	Subspecies	No.	Germplasm	Subspecies
1	C190	Indica	12	CG131R	Indica
2	CG133R	Indica	13	CG158R	Indica
3	HD63	Indica	14	CG111R	Indica
4	CG15R	Indica	15	CG114R	Indica
5	Javanica 22	Japonica	16	CG173R	Indica
6	Shuhui527	Indica	17	CG132R	Indica
7	3301R	Indica	18	CG151R	Indica
8	Lehui188	Indica	19	CG159R	Indica
9	Zhonghua9	Japonica	20	CG240R	Indica
10	Jing925	Japonica	21	BR207-2	Indica
11	BR207-1	Indica

Gene	Sequence of primers (5'-3')	Tm	Length for genome amplified (bp)	Chromosome
OsHKT1;1	F: GAGCACTGTGGAGGAATTTTACCG R: TAGTGAGTAGCCTACATTGCCGAAA	61.1	389	Chr.04
OsHKT1;3	F: GCTTACTTTGCCCTGATCTCCT R: TGAATACCTCACCACCAATC	58.0	214	Chr.02
OsHKT1;4	F: GCGACTCTGGCAAACTGATA R: GGTTCCTGTCTATGTGAAAATGAATA	57.5	211	Chr.04
OsHKT2;1	F: GTTAATTTTGTTGTTCTAGC R: ATGAGGCTGGAAAGTGTCAG	53.3	200	Chr.06
OsHKT2;3	F: CTGCCATGAGAAGGCGTACAA R: ATCGCATACTGATCGCTTCTGAT	59.2	152	Chr.01
OsHKT2;4	F: CTTGCCATGAGAAGCCATACAG R: CTTGATTCTTGCATAACATCATCA	57.6	146	Chr.06
OsHKT1;5	F: ACGACCCCATCAACTACAGCGTCC R: TGCTCCACTTCCCTGAGAAGCCAAC	65.3	833	Chr.01

Gene	Sequence of primers (5'-3')	Tm	Length for genome amplified (bp)	Chromosome
OsHKT1;1	F: GAGCACTGTGGAGGAATTTTACCG R: TAGTGAGTAGCCTACATTGCCGAAA	61.1	389	Chr.04
OsHKT1;3	F: GCTTACTTTGCCCTGATCTCCT R: TGAATACCTCACCACCAATC	58.0	214	Chr.02
OsHKT1;4	F: GCGACTCTGGCAAACTGATA R: GGTTCCTGTCTATGTGAAAATGAATA	57.5	211	Chr.04
OsHKT2;1	F: GTTAATTTTGTTGTTCTAGC R: ATGAGGCTGGAAAGTGTCAG	53.3	200	Chr.06
OsHKT2;3	F: CTGCCATGAGAAGGCGTACAA R: ATCGCATACTGATCGCTTCTGAT	59.2	152	Chr.01
OsHKT2;4	F: CTTGCCATGAGAAGCCATACAG R: CTTGATTCTTGCATAACATCATCA	57.6	146	Chr.06
OsHKT1;5	F: ACGACCCCATCAACTACAGCGTCC R: TGCTCCACTTCCCTGAGAAGCCAAC	65.3	833	Chr.01

No.	Average germination rate (%)		Relative salt damage rate (%)	Salt tolerance	Level
	CK	NaCl (1.5%)	Relative salt damage rate (%)
1	99.33±1.15 aA	36.67±0.01 cC	63.08	Weak	7
2	99.33±1.15 aA	7.78±1.92 ghFG	92.17	Very weak	9
3	83.33±0.01 gG	13.33±0.01 efgEFG	84.00	Very weak	9
4	60.20±0.34 lL	8.84±3.08 ghFG	85.97	Very weak	9
5	73.33±0.01 jJ	11.67±1.67 fghEFG	84.08	Very weak	9
6	99.33±1.15 aA	28.00±15.62 dCD	71.68	Weak	7
7	83.33±0.01 gG	14.00±6 efgEFG	83.20	Very weak	9
8	86.67±0.01 fF	13.34±8.80 efgEFG	84.60	Very weak	9
9	93.34±0.01 dD	15.00±5 efgEF	83.93	Very weak	9
10	90.00±0.02 eE	81.67±1.67 aA	9.26	Very strong	1
11	73.33±0.01 jJ	16.89±3.00 efDEF	71.96	Weak	7
12	93.34±0.02 dD	3.33±0.01 hG	96.43	Very weak	9
13	93.34±0.02 dD	26.67±3.34 dCD	71.43	Weak	7
14	80.00±0.02 hH	10.00±2 fghFG	87.50	Very weak	9
15	86.68±0.02 fF	21.67±1.67 deDE	75.00	Weak	7
16	66.68±0.02 kK	50.00±0.02 bB	25.02	Strong	3
17	98.02±0.02 bB	13.33±7.02 efgEFG	86.39	Very weak	9
18	96.67±0.01 cC	7.33±3.05 ghFG	92.41	Very weak	9
19	76.67±0.01 iI	8.89±5.09 fghFG	88.41	Very weak	9
20	99.33±1.15 aA	8.89±5.09 fghFG	91.09	Very weak	9
21	93.34±0.01 dD	16.89±3.06 efDEF	81.90	Very weak	9

不同水稻种质在不同生育期耐盐鉴定的差异

Identification of Salt Tolerance in Different Rice Germplasm at Different Growth Stages

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No.	NaCl concentration										Average
	0.3%		0.5%		0.7%		0.9%		1.1%		Average
	Relative seedling height	Relative root length	Relative seedling height	Relative root length	Relative seedling height	Relative root length	Relative seedling height	Relative root length	Relative seedling height	Relative root length	Relative seedling height	Relative root length
1	0.92	0.74	1.00	0.78	0.89	0.47	0.82	0.37	0.76	0.33	0.88	0.54
3	0.94	0.85	0.94	0.82	0.89	0.78	0.77	0.41	0.71	0.43	0.85	0.66
4	0.85	1.69	0.85	1.24	1.01	1.44	0.77	0.81	0.82	0.91	0.86	1.22
5	0.97	0.87	0.87	0.77	0.86	0.53	0.83	0.42	0.83	0.38	0.87	0.59
6	0.95	1.30	0.96	1.22	0.95	0.97	0.96	1.25	0.86	0.67	0.94	1.08
7	1.00	1.14	1.00	0.82	0.98	0.72	0.92	0.62	0.91	0.65	0.96	0.79
8	1.02	1.18	1.01	1.02	1.01	0.76	0.99	0.53	0.96	0.59	1.00	0.82
9	1.07	1.61	1.00	0.92	1.03	1.51	0.91	0.75	0.93	0.84	0.99	1.13
10	0.93	0.91	0.88	1.03	0.90	1.00	0.85	1.02	0.86	0.90	0.89	0.97
11	1.06	1.05	1.15	1.02	0.96	1.05	0.88	0.97	0.89	0.93	0.99	1.00
12	1.01	1.02	0.79	1.05	0.83	1.04	0.89	0.97	0.87	0.99	0.88	1.01
14	0.96	1.09	0.91	0.93	0.98	0.83	0.93	0.67	0.98	0.79	0.95	0.86
16	0.86	0.98	0.89	1.06	0.90	0.98	0.90	0.97	0.86	0.81	0.88	0.96
17	0.88	0.94	0.98	0.73	0.98	0.82	0.94	0.67	0.99	0.63	0.95	0.76
19	0.89	1.35	0.92	1.06	0.84	0.97	0.87	0.48	0.92	0.66	0.89	0.90
20	0.87	1.19	0.86	1.84	1.02	1.56	0.99	1.20	0.95	1.28	0.94	1.41
21	0.98	1.07	0.98	1.03	0.99	0.89	0.84	0.93	0.84	0.99	0.93	0.98

No.	NaCl concentration												Average
	0		0.3%		0.5%		0.7%		0.9%		1.1%		Average
	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortal- ity (%)
1	71.60	13.37	69.37	39.50	71.93	24.06	68.13	53.68	63.25	56.66	66.72	89.04	67.88 eE	52.59 qQ
3	72.54	37.44	68.31	57.27	67.33	65.58	64.50	89.79	65.15	90.05	61.47	94.35	65.35 hH	79.41 gG
4	65.21	43.64	60.9	86.26	62.25	97.12	63.94	93.69	54.00	100.00	58.90	100.00	60.02 kK	95.41 aA
5	58.99	32.17	58.32	49.48	44.24	61.40	45.09	81.67	49.40	99.20	41.81	100.00	47.77 pP	78.35 hH
6	65.96	28.14	55.80	64.53	54.95	59.88	55.27	78.24	53.96	91.49	64.07	93.80	56.81 lL	77.59 iI
7	51.76	34.30	59.98	56.32	50.71	84.40	48.66	86.47	51.56	100.00	63.01	100.00	54.78 oO	85.44 dD
8	61.72	15.12	51.85	42.46	53.95	87.83	42.69	84.49	41.18	100.00	42.93	100.00	46.52 qQ	82.96 fF
9	74.42	29.95	75.18	26.63	70.04	71.48	69.25	84.12	73.10	97.96	67.05	100.00	70.92 cC	76.04 kK
10	75.25	33.66	72.65	53.65	73.23	84.40	73.17	81.00	73.96	97.07	72.16	100.00	73.03 bB	83.22 eE
11	67.27	19.64	65.62	42.87	68.42	44.97	68.54	47.71	65.62	91.48	68.66	100.00	67.37 fF	61.41 oO
12	69.14	13.52	70.96	44.81	68.32	30.92	65.46	70.89	65.19	93.70	63.54	99.49	66.70 gG	67.96 mM
14	71.15	20.27	68.51	55.58	70.12	60.57	70.75	81.07	70.76	88.92	70.22	99.60	70.07 dD	77.15 jJ
16	61.67	14.11	64.32	50.08	63.70	87.71	65.50	95.07	63.77	100.00	61.30	100.00	63.72 iI	86.57 cC
17	52.96	4.87	55.11	75.41	58.78	90.19	56.18	100.00	53.77	100.00	56.23	100.00	56.02 mM	93.12 bB
19	76.00	40.14	75.79	54.37	75.74	73.99	75.14	74.93	71.70	81.24	70.60	84.52	73.79 aA	73.81 lL
20	60.01	33.42	55.99	79.19	50.96	53.49	59.31	83.03	59.75	50.41	52.32	71.36	55.67 nN	67.50 nN
21	58.84	18.89	64.62	14.35	67.90	47.86	64.07	42.02	61.10	92.95	58.82	100.00	63.30 jJ	59.44 pP

No.	NaCl concentration												Average
	0		0.3%		0.5%		0.7%		0.9%		1.1%		Average
	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortality (%)	Water content (%)	Leaf mortal- ity (%)
1	71.60	13.37	69.37	39.50	71.93	24.06	68.13	53.68	63.25	56.66	66.72	89.04	67.88 eE	52.59 qQ
3	72.54	37.44	68.31	57.27	67.33	65.58	64.50	89.79	65.15	90.05	61.47	94.35	65.35 hH	79.41 gG
4	65.21	43.64	60.9	86.26	62.25	97.12	63.94	93.69	54.00	100.00	58.90	100.00	60.02 kK	95.41 aA
5	58.99	32.17	58.32	49.48	44.24	61.40	45.09	81.67	49.40	99.20	41.81	100.00	47.77 pP	78.35 hH
6	65.96	28.14	55.80	64.53	54.95	59.88	55.27	78.24	53.96	91.49	64.07	93.80	56.81 lL	77.59 iI
7	51.76	34.30	59.98	56.32	50.71	84.40	48.66	86.47	51.56	100.00	63.01	100.00	54.78 oO	85.44 dD
8	61.72	15.12	51.85	42.46	53.95	87.83	42.69	84.49	41.18	100.00	42.93	100.00	46.52 qQ	82.96 fF
9	74.42	29.95	75.18	26.63	70.04	71.48	69.25	84.12	73.10	97.96	67.05	100.00	70.92 cC	76.04 kK
10	75.25	33.66	72.65	53.65	73.23	84.40	73.17	81.00	73.96	97.07	72.16	100.00	73.03 bB	83.22 eE
11	67.27	19.64	65.62	42.87	68.42	44.97	68.54	47.71	65.62	91.48	68.66	100.00	67.37 fF	61.41 oO
12	69.14	13.52	70.96	44.81	68.32	30.92	65.46	70.89	65.19	93.70	63.54	99.49	66.70 gG	67.96 mM
14	71.15	20.27	68.51	55.58	70.12	60.57	70.75	81.07	70.76	88.92	70.22	99.60	70.07 dD	77.15 jJ
16	61.67	14.11	64.32	50.08	63.70	87.71	65.50	95.07	63.77	100.00	61.30	100.00	63.72 iI	86.57 cC
17	52.96	4.87	55.11	75.41	58.78	90.19	56.18	100.00	53.77	100.00	56.23	100.00	56.02 mM	93.12 bB
19	76.00	40.14	75.79	54.37	75.74	73.99	75.14	74.93	71.70	81.24	70.60	84.52	73.79 aA	73.81 lL
20	60.01	33.42	55.99	79.19	50.96	53.49	59.31	83.03	59.75	50.41	52.32	71.36	55.67 nN	67.50 nN
21	58.84	18.89	64.62	14.35	67.90	47.86	64.07	42.02	61.10	92.95	58.82	100.00	63.30 jJ	59.44 pP

Primers	1-C190	4-CG15R	8-Lehui188	10-Jing925	11-BR207-1	16-CG173R	19-CG159R	20-CG240R
OsHKT1;1	1	1	1	1	1	1	1	1
OsHKT1;3	1	1	1	1	1	1	1	1
OsHKT1;4	1	1	1	1	1	1	1	1
OsHKT1;5	2	2	2	2	2	2	2	2
OsHKT2;1	1	1	1	1	1	1	1	1
OsHKT2;3	1	1	1	1	1	1	1	1
OsHKT2;4	0	0	0	0	0	0	0	0

Primers	1-C190	4-CG15R	8-Lehui188	10-Jing925	11-BR207-1	16-CG173R	19-CG159R	20-CG240R
OsHKT1;1	1	1	1	1	1	1	1	1
OsHKT1;3	1	1	1	1	1	1	1	1
OsHKT1;4	1	1	1	1	1	1	1	1
OsHKT1;5	2	2	2	2	2	2	2	2
OsHKT2;1	1	1	1	1	1	1	1	1
OsHKT2;3	1	1	1	1	1	1	1	1
OsHKT2;4	0	0	0	0	0	0	0	0