拟南芥线粒体蛋白突变体ssr1-2表型的详细鉴定与分析

doi:10.7525/j.issn.1673-5102.2023.03.012

摘要/Abstract

摘要：

SSR1在拟南芥（Arabidopsis thaliana）中编码一个线粒体蛋白，为深入了解SSR1基因在植物生长发育和逆境应答中的作用，以ssr1-2及其抑制子突变体EMS143和EMS145为材料，对其根和地上部分的生长，及其对脯氨酸的敏感性和铁稳态进行跟踪分析，并利用拟南芥幼苗嫁接技术分析了ssr1-2短根表型对地上部分生长的影响。结果表明：ssr1-2主根长变短，根系的形态与须根系类似。地上部分也小于野生型，但出现得晚于根短表型。嫁接试验结果表明，突变体的根能限制野生型的地上部分生长，反之突变体的地上部分也能影响野生型的根，但前者的作用更大。ssr1-2在种子萌发、根长和叶绿素含量等指标上表现出对脯氨酸的超敏感表型。此外，ssr1-2对铁营养不敏感，即铁盐对其生长的促进作用显著小于WS野生型，说明其利用铁的能力显著下降。这些结果表明SSR1可能通过影响铁营养利用参与拟南芥根生长的调控，暗示线粒体铁元素利用机制的损伤可能是脯氨酸对植物生长发育抑制作用加强的原因之一。

关键词: SSR1, 脯氨酸处理, 铁硫簇合成, 嫁接, 拟南芥

Abstract:

To explore the role of SSR1 gene encoding a functionally unknown mitochondrial protein of Arabidopsis thaliana in plant growth and stress response， ssr1-2 and its suppressor mutants EMS143 and EMS145 were used to track the growth of their roots and aboveground parts， as well as their proline sensitivity and iron homeostasis， and the effects of ssr1-2 short root phenotype on aboveground growth of seedlings were analyzed by micrografting. The results showed that the primary root length of ssr1-2 was shorter and the root architecture was similar to the fibrous root system. The growth of their shoots was also retarded， but appeared considerably later than the short-root phenotype. The results of micrografting revealed that the roots of ssr1-2 could restrict the growth of the wild-type shoots and， and conversely， the shoot of ssr1-2 could also affect the growth of the wild-type roots， but the former had a bigger effect. ssr1-2 showed a hypersensitive phenotype to proline in seed germination， root length and leaf chlorophyll content. In addition， ssr1-2 appeared insensitive to Fe nutrition， that was， the stimulating effect of Fe salt on seedling growth was significantly less than that of wild-type WS， indicating a significant decrease in its ability to utilize Fe. The results suggested that SSR1 might be involved in the regulation of A. thaliana root growth by affecting the utilization of iron nutrient， implying that the impairment of the mitochondrial iron utilization machinery might contribute significantly to the enhanced inhibitory effect of proline on plant growth and development.

Key words: SSR1, proline treatment, iron-sulfur cluster assembly, micrografting, Arabidopsis thaliana

中图分类号:

S335

蔡圆圆, 夏季奔奔, 应文涵, 王洁瑶, 谢涛, 邢孔丫, 冯宣军, 华学军. 拟南芥线粒体蛋白突变体ssr1-2表型的详细鉴定与分析[J]. 植物研究, 2023, 43(3): 421-431.

Yuanyuan CAI, Jibenben XIA, Wenhan YING, Jieyao WANG, Tao XIE, Kongya XING, Xuanjun FENG, Xuejun HUA. Detailed Phenotypical Analysis on the Mutant ssr1-2 Encoding a Mitochondrial Protein of Arabidopsis thaliana[J]. Bulletin of Botanical Research, 2023, 43(3): 421-431.

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时间 Time /d	相对萌发率 Relative germination rate /%
时间 Time /d	WS	ssr1-2	ssr1-2C	EMS143	EMS145
1	74.80	31.06	79.08	63.11	108.50
2	98.95	96.21	98.91	121.44	100.50
3	99.42	100.52	94.07	99.50	98.89
4	99.22	98.11	89.17	105.07	95.98
5	101.05	98.13	93.24	106.63	98.50
6	101.95	101.01	95.65	111.17	96.48
7	101.23	100.29	95.65	111.17	95.50

生长条件 Growth condition	株系 Line	指标Index
生长条件 Growth condition	株系 Line	主根长 Primary foot length /cm	叶片面积 Leaf area /cm²	叶绿素质量分数 Chlorophyll content /mg·g^-1
MS培养基 MS medium	WS	2.13	0.61	0.72
	ssr1-2	0.37	0.61	0.65
	ssr1-2C	2.16	0.63	0.73
	EMS143	2.01	0.70	0.57
	EMS145	1.98	0.66	0.67
20 mmol·L^-1 Pro的MS培养基 MS medium containg 20 mmol·L^-1 Pro	WS	1.00	0.43	0.27
	ssr1-2	0.16	0.37	0.17
	ssr1-2C	1.04	0.43	0.27
	EMS143	0.44	0.44	0.27
	EMS145	0.42	0.53	0.40

指标 Index	株系 Line	处理条件 Treat condition
指标 Index	株系 Line	处理1 Treat 1	处理2 Treat 2	处理3 Treat 3	处理4 Treat 4	处理5 Treat 5
主根长 Primary foot length /cm	WS	0.61	2.49	2.09	1.90	1.41
	ssr1-2	0.47	0.46	0.46	0.49	0.46
	ssr1-2C	0.63	2.44	1.99	1.95	1.46
	EMS143	0.60	1.90	1.82	1.98	1.68
	EMS145	0.45	1.18	1.25	0.88	0.65
净生长量 Net growth of primary root /cm	WS	0.04	0.03	0.39	0.22	0.35
	ssr1-2	0.06	0.02	0.03	0.01	0.01
	ssr1-2C	0.05	0.03	0.31	0.25	0.29
	EMS143	0.04	0.03	0.26	0.34	0.31
	EMS145	0.03	0.01	0.12	0.16	0.13
侧根长 Lateral root length /cm	WS	0.23	0.45	0.69	0.60	0.41
	ssr1-2	0.29	0.19	0.22	0.25	0.20
	ssr1-2C	0.23	0.45	0.68	0.64	0.42
	EMS143	0.21	0.81	0.80	1.03	0.75
	EMS145	0.20	0.44	0.78	0.81	0.68
侧根密度 Lateral root density/（number·cm^-1）	WS	3.22	1.72	1.91	1.89	2.09
	ssr1-2	4.44	4.67	4.83	4.67	4.10
	ssr1-2C	3.47	1.74	1.89	1.84	2.00
	EMS143	1.71	2.03	3.37	1.64	1.95
	EMS145	2.82	2.19	2.93	3.94	3.23
叶片面积 Leaf area /cm²	WS	0.72	0.73	0.73	0.75	0.75
	ssr1-2	0.67	0.72	0.66	0.63	0.62
	ssr1-2C	0.74	0.76	0.75	0.78	0.77
	EMS143	0.52	0.67	0.67	0.68	0.69
	EMS145	0.52	0.68	0.70	0.67	0.67
叶绿素质量分数 Chlorophyll content /（mg·g^-1）	WS	0.51	1.17	1.12	1.20	1.26
	ssr1-2	0.69	0.98	1.20	0.97	1.07
	ssr1-2C	0.53	1.22	1.11	1.16	1.19
	EMS143	0.41	0.97	0.99	0.94	0.94
	EMS145	0.34	1.13	1.11	0.91	0.94

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