水稻白条纹叶突变体 st11的遗传分析与基因定位

doi:10.3969/j.issn.1001-7216.2015.01.002

摘要/Abstract

摘要：

白条纹叶突变体 st11是从粳稻品种Kitaake组培过程中获得的。该突变体在分蘖前叶色表现为正常,从分蘖期开始新生叶表现为白条纹直至成熟期。与野生型相比,该突变体的分蘖、株高、结实率和千粒重等农艺性状没有发生明显变化。遗传分析表明该突变体白条纹叶性状受一对隐性核基因控制。利用该突变体分别与水稻02428、Jodan杂交构建了两个F₂群体用于基因定位。通过集群分离分析(bulked segregant analysis)发现该基因位于第1染色体端粒附近,并与分子标记RM151和RM10080连锁。进一步利用更多分子标记分析F₂群体,我们将该基因定位于I10和I26两个标记之间大约270kb的区间内。

关键词: 水稻, 白条纹叶突变体, 分子标记, 基因定位

Abstract:

A white stripe leaf mutant st11 was obtained from tissue-cultured rice Kitaake (Oryza sativa spp japonica). This mutant was characterized by white stripe leaves from tillering stage to mature stage. Compared with the wild type Kitaake, the mutant showed no obvious changes in agricultural traits, such as tiller number, plant height, seed-setting rate, and grain weight. Genetic analyses showed that the white stripe leaf phenotype in the mutant st11 was controlled by a single recessive gene. Two F₂ segregating populations derived from the crosses, st11×Jodan and st11×02428, respectively, were used for mapping the gene st11. Bulked segregant analysis suggested that the gene st11 was located close to the markers, RM151 and RM10080, on chromosome 1. Using more molecular markers, we finally delimited the gene st11 to a region of about 270 kb between the Indel markers, I10 and I26, on the telomere of the short arm of chromosome 1.

Key words: rice, white stripe leaf mutant, molecular marker, gene mapping

中图分类号:

Q343.5
S754

成钦淑, 叶邦全, 袁灿, 李伟滔, 尹俊杰, 王静, 贺闽, 汪吉春, 王玉平, 李仕贵, 陈学伟. 水稻白条纹叶突变体 st11的遗传分析与基因定位[J]. 中国水稻科学, 2015, 29(1): 14-21.

Qin-shu CHENG, Bang-quan YE, Can YUAN, Wei-tao LI, Jun-jie YIN, Jing WANG, Min HE, Ji-chun WANG, Yu-ping WANG, Shi-gui LI, Xue-wei CHEN. Genetic Analysis and Gene Mapping of White Stripe Leaf Mutant st11 in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(1): 14-21.

图/表 9

表1 定位基因 st11用到的部分重要引物

Table 1 Some important primer pairs used for gene mapping.

分子标记 Marker	正向引物 Forward primer	反向引物 Reverse primer	物理距离^a Distance/Mb^a
I26	ATTCAGAAGGTGATGCTCCG	GATATGTCGCCAGAGACCCT	0.188
I10	TGCAACGACGGGCATTTTG	GAGCTTCTTGATGTAGGCCG	0.455
I14	GGGGCTCAACATGGACCAAA	GGCATGTTGAAGCTGAGCAC	0.689
RM10048	CAAGCAGTGATCATACAGCCTTCC	GCCATGGCTGAGAACAGAGAGC	0.744
RM10076	CTAGCAGCTGTCTGCGACACACG	CCGAGGTGTTATGCCAATCTCTATGG	1.351

图1 野生型WT和突变体 st11不同时期叶色表型 A-苗期; B-分蘖期; C-分蘖期叶片; WT-野生型; s11-突变体。下同。

Fig. 1. Photographs of the representative plants and leaves from the Kitaake (WT) and mutant st11 plants. A, At seedling stage; B, At tillering stage; C, Leaves during tillering stage; WT, Wild type; st11,Mutant. The same as below.

图2 野生型(WT)和突变体(st11)间主要农艺性状比较分析

Fig. 2. Comparative analyses of main agronomic traits between the wild type (WT) and the mutant (st11).

图3 突变体 st11白化条斑性状与潮霉素共分离分析 1-潮霉素阳性单株对照; 2-潮霉素阴性单株对照; 3~24-表型为白条纹的22个F2单株。

Fig. 3. Co-segregation analysis of the white stripe leaf phenotype with the gene hygromycin on the F2 mutant plants derived from the cross of Kitaake× st11. 1, Positive control of hygromycin; 2, Negative control of hygromycin; 3-24, Twenty-two F2 plants with the white stripe leaf phenotype.

表2 突变体 st11的遗传分析

Table 2 Genetic analysis of the white stripe leaf phenotype of the mutant st11.

组合 Cross	F₁	F₂			X²_(3:1)
组合 Cross	F₁	总株数 Total no. of plants	正常植株 No. of normal plants	突变体植株 No. of white strip plants	X²_(3:1)
st11/Jodan	正常绿叶 Normal green leaf	1035	791	244	0.589
st11/02428	正常绿叶 Normal green leaf	599	453	146	0.054
st11/C418	正常绿叶 Normal green leaf	448	343	105	0.502
C418/st11	正常绿叶 Normal green leaf	421	324	97	0.863

图4 标记I10在22个F2白条斑突变单株的分离情况 1~22表示来源于亲本Jodan与 st11构建的F2群体的22个不同单株;0表示亲本Jodan。

Fig. 4. Genotype analysis of the 22 F2 plants with the white stripe leaf phenotype using the indel marker I10. 1-22 represent 22 individual plants derived from the cross of Jodan×Kitaake; 0 represent the parent Jodan.

表3 五个重要分子标记在 st11/ Jodan的F2群体中白条纹单株的交换情况

Table 3 Recombinant events of five molecular makers in the plants with white stripe leaf from F2 population of st11/Jodan.

交换单株号 No. of recombinant plants	标记 Marker
交换单株号 No. of recombinant plants	I26	I10	I14	RM10048	RM10076
46	1	0	0	0	0
51	1	0	0	0	0
84	1	0	0	0	0
24	0	0	1	1	1
81	0	0	1	1	1
93	0	0	1	1	1
23	0	0	0	0	2
41	0	0	0	0	1
44	0	0	0	0	1
60	0	0	0	0	1
74	0	0	0	0	1
81	0	0	0	0	1
84	0	0	0	0	1
107	0	0	0	0	1
110	0	0	0	0	1
112	0	0	0	0	1
115	0	0	0	0	1
165	0	0	0	0	1

图5 基因 st11在水稻第1染色体上的分子定位 A-利用 st11/Jodan的F2群体构建的st11的分子定位结果; B-利用st11/02428的F2群体构建的st11的分子定位结果; C-结合图A和图B的分子定位结果获得的st11的分子定位。N1-由亲本 st11和Jodan构建的F2群体中白条纹单株; N2-由亲本 st11和02428构建的F2群体中的白条纹单株。

Fig. 5. Molecular mapping of gene st11 on chromosome 1. A, Molecular mapping of st11based on F2 population derived from the cross of st11/Jodan; B, Molecular mapping of st11based on F2 population derived from the cross of st11/02428; C, Molecular mapping of st11based on the results from A and B. N1, Plants with the white stripe leaf phenotype in F2 population derived from the cross of st11/Jodan; N2, plants with the white stripe leaf phenotype in F2 population derived from the cross of st11/02428.

表4 两个重要分子标记在 st11/ 02428的F2群体中白条纹单株的交换情况

Table 4 Recombinant events of two molecular makers in the plants with white stripe leaf from F2 population of st11/02428.

交换单株号 No. of recombinant plants	引物 Marker		交换单株号 No. of recombinant plants	引物 Marker
交换单株号 No. of recombinant plants	I10	RM10076	交换单株号 No. of recombinant plants	I10	RM10076
1	0	1	249	0	1
2	0	1	256	0	1
10	0	1	260	0	1
36	0	1	266	0	1
39	0	1	269	0	1
55	0	1	287	0	1
59	0	1	288	1	1
60	0	1	297	0	1
63	1	1	299	0	1
67	0	1	317	0	1
68	1	1	320	0	1
90	0	1	322	0	1
93	0	1	332	0	1
97	0	1	338	0	1
102	0	1	340	0	1
129	0	1	344	0	1
136	0	1	353	0	1
141	0	1	361	0	1
172	0	1	372	0	1
183	0	1	379	0	1
190	0	1	382	0	1
193	0	1	385	0	1
199	0	1	388	0	1
209	0	1	402	0	1
211	0	1	409	0	1
221	0	1	412	0	1
222	1	2	414	0	1
228	0	1	420	0	1
236	0	1	429	0	1
238	0	1	441	0	1
246	0	1	445	0	1
248	0	1	450	0	1
			464	0	1

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