Development and Application of a Functional Marker for the Cold Stress Responsive Gene OsSADMC

doi:10.3969/j.issn.1001G7216.2015.05.004

Abstract

Abstract:

Cold tolerance of 95 rice germplasms were screened at seedling stage. Among them, ten cold tolerant lines (survival rate ≥60%) and nine cold sensitive lines (survival rate=0%) were identified. Gene expression analysis of cold stress responsive gene OsSADMC showed that the expression levels of OsSADMC were significantly higher in the cold tolerant lines than those in the cold sensitive lines at both normal and cold stress conditions. Ten SNP loci were identified in the gene coding region between Lijiangxintuanheigu(LTH) and IR29. One amino acid mutation (from serine to leucine) was found between LTH and IR29. A functional marker (SADMC-CAPS1) was developed based on one SNP locus. The allele of OsSADMC carried by ten cold tolerant lines could be distinctly distinguished from that of nine cold sensitive lines by this functional marker.

Key words: rice, cold tolerance, functional marker, OsSADMC

摘要：

通过对95份水稻种质资源进行苗期耐冷性鉴定,筛选出10份苗期耐冷(存活率≥60%)和9份冷敏感的材料(存活率为零),并对水稻冷胁迫响应基因OsSADMC进行了表达分析,发现对照和冷胁迫处理条件下该基因在耐冷品种中的表达量都显著高于冷敏感品种。对耐冷材料丽江新团黑谷(LTH)和冷敏感材料IR29的基因编码区域进行了克隆和测序分析,在两个品种间鉴定了10个SNP位点,其中第749碱基处存在的一个碱基变异(C突变为T),从而使丝氨酸变为亮氨酸。根据其中一个SNP位点设计了OsSADMC的功能标记SADMC-CAPS1,该标记可以准确地鉴定出19份耐冷性不同水稻种质资源OsSADMC的基因型和耐冷性。

关键词: 水稻, 耐冷性, 功能标记, OsSADMC

CLC Number:

Xiao-chen HU, Ting ZHANG, Sheng YANG, Qiao QIN, Ying-yao SHI, Wen-sheng WANG, Bin-ying FU. Development and Application of a Functional Marker for the Cold Stress Responsive Gene OsSADMC[J]. Chinese Journal OF Rice Science, 2015, 29(5): 475-480.

胡晓晨, 张婷, 杨圣, 秦巧, 石英尧, 王文生, 傅彬英. 水稻冷胁迫响应基因OsSADMC功能标记的开发和利用[J]. 中国水稻科学, 2015, 29(5): 475-480.

Figures/Tables 4

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品种编号 Variety code	品种名称 Variety name	类型 Type	总株数 No of plants	平均存活率 Average survival rate /%	耐冷性 Cold tolerance
22	IR9814-11-3	籼稻indica	30	0	冷敏感 Sensitive
31	CSR9	籼稻indica	30	0	冷敏感 Sensitive
39	黄金早Huangjinzao	籼稻indica	30	0	冷敏感 Sensitive
41	寒赤Hanchi	籼稻indica	30	0	冷敏感 Sensitive
48	MRC8009-3536	籼稻indica	30	0	冷敏感 Sensitive
70	OR410-7	籼稻indica	30	0	冷敏感 Sensitive
89	Kasalath	籼稻indica	30	0	冷敏感 Sensitive
92	9311	籼稻indica	30	0	冷敏感 Sensitive
94	IR29	籼稻indica	30	0	冷敏感 Sensitive
5	C82-27	粳稻 japonica	30	63	耐冷 Tolerant
7	Meqamunin (2)	粳稻 japonica	30	60	耐冷 Tolerant
8	Leuwrro	粳稻 japonica	30	63	耐冷 Tolerant
9	Litchikiang	粳稻 japonica	30	67	耐冷 Tolerant
11	Reishiko	粳稻 japonica	30	60	耐冷 Tolerant
15	Basmati 410	籼稻indica	30	63	耐冷 Tolerant
82	越光Koshihikari	粳稻japonica	30	63	耐冷 Tolerant
86	丽江新团黑谷 Lijiangxintuanheigu	粳稻 japonica	30	73	耐冷 Tolerant
87	K354	粳稻 japonica	30	60	耐冷 Tolerant
91	松粳9 Songjing 9	粳稻 japonica	30	67	耐冷 Tolerant

品种编号 Variety code	品种名称 Variety name	类型 Type	总株数 No of plants	平均存活率 Average survival rate /%	耐冷性 Cold tolerance
22	IR9814-11-3	籼稻indica	30	0	冷敏感 Sensitive
31	CSR9	籼稻indica	30	0	冷敏感 Sensitive
39	黄金早Huangjinzao	籼稻indica	30	0	冷敏感 Sensitive
41	寒赤Hanchi	籼稻indica	30	0	冷敏感 Sensitive
48	MRC8009-3536	籼稻indica	30	0	冷敏感 Sensitive
70	OR410-7	籼稻indica	30	0	冷敏感 Sensitive
89	Kasalath	籼稻indica	30	0	冷敏感 Sensitive
92	9311	籼稻indica	30	0	冷敏感 Sensitive
94	IR29	籼稻indica	30	0	冷敏感 Sensitive
5	C82-27	粳稻 japonica	30	63	耐冷 Tolerant
7	Meqamunin (2)	粳稻 japonica	30	60	耐冷 Tolerant
8	Leuwrro	粳稻 japonica	30	63	耐冷 Tolerant
9	Litchikiang	粳稻 japonica	30	67	耐冷 Tolerant
11	Reishiko	粳稻 japonica	30	60	耐冷 Tolerant
15	Basmati 410	籼稻indica	30	63	耐冷 Tolerant
82	越光Koshihikari	粳稻japonica	30	63	耐冷 Tolerant
86	丽江新团黑谷 Lijiangxintuanheigu	粳稻 japonica	30	73	耐冷 Tolerant
87	K354	粳稻 japonica	30	60	耐冷 Tolerant
91	松粳9 Songjing 9	粳稻 japonica	30	67	耐冷 Tolerant

引物名称 Primer name	引物序列(5'-3') Primer sequence (5'-3')	酶切位点 Restriction site	酶切产物大小 Size of enzyme-digested product/bp
SAMDC-CAPs1F	TCTGAGTCCAGCCTGTTTATCT	CCGCGG	351;211/140
SAMDC-CAPs1R	TCATGCACATTTCAAGGGTAA	GGCGCC

引物名称 Primer name	引物序列(5'-3') Primer sequence (5'-3')	酶切位点 Restriction site	酶切产物大小 Size of enzyme-digested product/bp
SAMDC-CAPs1F	TCTGAGTCCAGCCTGTTTATCT	CCGCGG	351;211/140
SAMDC-CAPs1R	TCATGCACATTTCAAGGGTAA	GGCGCC