Identification and Fine Mapping of a Narrow Leaf Mutant nal10 in Rice

doi:10.3969/j.issn.1001G7216.2015.06.004

Chinese Journal OF Rice Science ›› 2015, Vol. 29 ›› Issue (6): 587-594.DOI: 10.3969/j.issn.1001G7216.2015.06.004

• Orginal Article • Previous Articles Next Articles

Identification and Fine Mapping of a Narrow Leaf Mutant nal10 in Rice

Yun-xia FANG^1,^2,³, Li ZHU¹, Jiang-jie PAN⁴, Hai-ping YU², Da-wei XUE⁴, Yu-chun RAO², Xiao-hu WANG², Zhen-yuan SHI², Dong ZHANG⁴, yang-zhou ZHU², Guo-jun DONG², Da-li ZENG², Guang-heng ZHANG², Long-biao GUO², Jiang HU^2,^*(), Qian QIAN^2,^*()

¹College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China
Rice Research Institute, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China
College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China

Received:2015-01-30 Revised:2015-03-17 Online:2015-10-25 Published:2015-11-10
Contact: Jiang HU, Qian QIAN
About author:
^*Corresponding author：E-mail:qianqian188@hotmail.com,hujiang588@163.com

水稻窄叶突变体nal10的鉴定与基因精细定位

方云霞^1,^2,³, 朱丽², 潘江杰⁴, 余海平², 薛大伟⁴, 饶玉春², 王小虎², 石珍源², 张栋⁴, 祝阳舟², 董国军², 曾大力², 张光恒², 郭龙彪², 胡江^2,^*(), 钱前^2,^*()

¹浙江大学农业与生物技术学院, 杭州310058
²中国水稻研究所水稻生物学国家重点实验室, 杭州310006江西省农业科学院水稻研究所, 南昌 330200
³江西省农业科学院水稻研究所, 南昌 330200
⁴杭州师范大学生命与环境学院,杭州310036

通讯作者: 胡江,钱前
作者简介:
^*通讯录作者：E-mail:qianqian188@hotmail.com,hujiang588@163.com
基金资助:
国家自然科学基金资助项目(31201194,91335105)

Abstract

Abstract:

A novel rice mutant narrow leaf 10 (nal10) was identified from an EMS (ethyl methane sulphonate) mutagenized population of Wuyunjing 7. The mutant was characterized by narrow leaves and smaller architecture phenotype at the seedling stage, and dwarf, narrow leaves, deformed flowers and male sterility at the reproduction stage. Cyto-anatomical analysis indicated that the mutant had fewer large and small veins compared with the wild-type, which may account for the narrow leaf phenotype. The genetic analysis indicated the narrow leaf trait was controlled by a recessive nuclear gene. Using F₂ population derived from the cross of nal10 and Taizhongbendi 1, three closely linked SSR markers RM1287, RM562 and RM5638 on chromosome 1 were obtained by the bulked segregant analysis method. With the developed new markers, the gene was finally fine-mapped to an interval of 55.2 kb between the STS markers PK83 and PK78, where a total of eight ORFs were found. RT-PCR analysis indicated the expression levels of auxin biosynthesis, signal transduction and transport related genes were all down-regulated, suggesting NAL10 is a new narrow leaf gene involving in the auxin metabolism. These results laid a foundation for further cloning of the gene and enrich the genetic regulation network of leaf development in rice.

Key words: rice, narrow leaf, NAL10, gene mapping

摘要：

利用EMS诱变粳稻品种武运粳7号获得一个新窄叶突变体nal10,该突变体在苗期表现出极窄叶和弱小的特征,在生殖期表现出矮化、窄叶、畸颖和不育的表型。组织解剖学分析表明,nal10的叶片窄化可能是由于叶片大脉和小脉数量减少造成的。遗传分析表明,该窄叶表型受一对隐性核基因控制。利用nal10与台中本地一号(TN1)构建的F₂群体,通过混合分离法,在第1染色体上找到3个紧密连锁的SSR标记(RM1287、RM562和RM5638)。进一步利用新发展的分子标记,最终将该基因定位在STS标记PK83和PK78之间的55.2 kb范围内,该区间共有8个开放阅读框。RT-PCR分析表明,NAL10的突变能够造成生长素生物合成、信号转导和运输基因转录水平的下降,因此NAL10是一个与生长素相关的窄叶新基因。这些结果为进一步克隆该基因、丰富水稻叶发育的遗传调控网络打下了基础。

关键词: 水稻, 窄叶, NAL10, 基因定位

CLC Number:

Yun-xia FANG, Li ZHU, Jiang-jie PAN, Hai-ping YU, Da-wei XUE, Yu-chun RAO, Xiao-hu WANG, Zhen-yuan SHI, Dong ZHANG, yang-zhou ZHU, Guo-jun DONG, Da-li ZENG, Guang-heng ZHANG, Long-biao GUO, Jiang HU, Qian QIAN. Identification and Fine Mapping of a Narrow Leaf Mutant nal10 in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(6): 587-594.

方云霞, 朱丽, 潘江杰, 余海平, 薛大伟, 饶玉春, 王小虎, 石珍源, 张栋, 祝阳舟, 董国军, 曾大力, 张光恒, 郭龙彪, 胡江, 钱前. 水稻窄叶突变体nal10的鉴定与基因精细定位[J]. 中国水稻科学, 2015, 29(6): 587-594.

Figures/Tables 7

References 30

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分子标记 Marker	正向引物序列(5'-3') Forward primer sequence (5'-3')	反向引物序列(5'-3') Reverse primer sequence (5'-3')	实验目的 Purpose
PK18	CATAAGGACGAACCAGAGGC	CATATCACCACCCACTAGCAA	Mapping
PK20	GGAACTGCATTTCGATTCATAC	TACTCGCTACAAGACATCACAA	Mapping
PK25	GTAGGTCGTCCTGATACTCGG	TACTTGTATCTTGCCCTATGCT	Mapping
PK30	CAAGTAACTCTTTGGTCATGCA	GCATTTCCCATTGACTGAAC	Mapping
PK37	CGCATTGTCAAGATGAGGAAG	TTAGACCACCCCTAAAGCAAT	Mapping
PK49	CTTATGAATCCTGAAGCCACA	CCTTAACCATTGATTGTCCC	Mapping
PK66	ATCAGTTAGAACCACGAAGCA	GCCTTTTGATATCTTCAGCG	Mapping
PK76	GCAGCTCGTGGACACTCGTC	CCCTGCGTTATCCCTCTTCC	Mapping
PK78	CATAGGCCATGCATTGACAGA	AGGTGGGGAAAAGGTAAAACTG	Mapping
PK83	CAAAAACTTATATCAATGCG	GCTTGTAAATATACATTGTCGTC	Mapping
PIN1	CGTCTGCTTCAGGTGGAACTT	GACAGGATCGAGATGGATTTCAG	qRT-PCR
LAX1	TTGCCTTGCTGTGTCTTCGT	TGGACATTGCACACCGAGTAG	qRT-PCR
AGAP	CTGACCCTTGGGCAACAGA	GGGTTCAGGTTGGCTTTGATC	qRT-PCR
IAA3	CCCGTGGCAGATGTTTGTC	TTCGGAACCCTTCATGATCCT	qRT-PCR
IAA31	TTCGTGAAGGTGAGCATGGA	CGGTAGCCCTTGTACACCTTGA	qRT-PCR
YUCCA1	CGCAACACGGTGCATGTC	CGATGCCGAACGTGGATAG	qRT-PCR
RCN1	TCATCAACCGCGACAGGAT	CCGCCTCGTACGGGTACTT	qRT-PCR
Actin	ATCCATCTTGGCATCTCTCAGC	CACAATGGATGGGCCAGACT	qRT-PCR

分子标记 Marker	正向引物序列(5'-3') Forward primer sequence (5'-3')	反向引物序列(5'-3') Reverse primer sequence (5'-3')	实验目的 Purpose
PK18	CATAAGGACGAACCAGAGGC	CATATCACCACCCACTAGCAA	Mapping
PK20	GGAACTGCATTTCGATTCATAC	TACTCGCTACAAGACATCACAA	Mapping
PK25	GTAGGTCGTCCTGATACTCGG	TACTTGTATCTTGCCCTATGCT	Mapping
PK30	CAAGTAACTCTTTGGTCATGCA	GCATTTCCCATTGACTGAAC	Mapping
PK37	CGCATTGTCAAGATGAGGAAG	TTAGACCACCCCTAAAGCAAT	Mapping
PK49	CTTATGAATCCTGAAGCCACA	CCTTAACCATTGATTGTCCC	Mapping
PK66	ATCAGTTAGAACCACGAAGCA	GCCTTTTGATATCTTCAGCG	Mapping
PK76	GCAGCTCGTGGACACTCGTC	CCCTGCGTTATCCCTCTTCC	Mapping
PK78	CATAGGCCATGCATTGACAGA	AGGTGGGGAAAAGGTAAAACTG	Mapping
PK83	CAAAAACTTATATCAATGCG	GCTTGTAAATATACATTGTCGTC	Mapping
PIN1	CGTCTGCTTCAGGTGGAACTT	GACAGGATCGAGATGGATTTCAG	qRT-PCR
LAX1	TTGCCTTGCTGTGTCTTCGT	TGGACATTGCACACCGAGTAG	qRT-PCR
AGAP	CTGACCCTTGGGCAACAGA	GGGTTCAGGTTGGCTTTGATC	qRT-PCR
IAA3	CCCGTGGCAGATGTTTGTC	TTCGGAACCCTTCATGATCCT	qRT-PCR
IAA31	TTCGTGAAGGTGAGCATGGA	CGGTAGCCCTTGTACACCTTGA	qRT-PCR
YUCCA1	CGCAACACGGTGCATGTC	CGATGCCGAACGTGGATAG	qRT-PCR
RCN1	TCATCAACCGCGACAGGAT	CCGCCTCGTACGGGTACTT	qRT-PCR
Actin	ATCCATCTTGGCATCTCTCAGC	CACAATGGATGGGCCAGACT	qRT-PCR

基因 Locus name	功能注释 Function annotation
LOC_Os01g34580	Hypothetical protein
LOC_Os01g34590	Expressed protein
LOC_Os01g34600	Expressed protein
LOC_Os01g34610	Expressed protein
LOC_Os01g34614	DNA directed RNA polymerase, 7 kDa subunit domain containing protein, expressed
LOC_Os01g34620	OsGrx_S15.1 - glutaredoxin subgroup II, expressed
LOC_Os01g34630	Retrotransposon protein, putative, Ty1-copia subclass, expressed
LOC_Os01g34660	RNAse P Rpr2/Rpp21 subunit domain containing protein, fexpressed

基因 Locus name	功能注释 Function annotation
LOC_Os01g34580	Hypothetical protein
LOC_Os01g34590	Expressed protein
LOC_Os01g34600	Expressed protein
LOC_Os01g34610	Expressed protein
LOC_Os01g34614	DNA directed RNA polymerase, 7 kDa subunit domain containing protein, expressed
LOC_Os01g34620	OsGrx_S15.1 - glutaredoxin subgroup II, expressed
LOC_Os01g34630	Retrotransposon protein, putative, Ty1-copia subclass, expressed
LOC_Os01g34660	RNAse P Rpr2/Rpp21 subunit domain containing protein, fexpressed

Identification and Fine Mapping of a Narrow Leaf Mutant nal10 in Rice

水稻窄叶突变体nal10的鉴定与基因精细定位

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