水稻类病斑突变体lmm4的鉴定及其基因定位

邱结华1 ,马宁1, 2,蒋汉伟1, 2 ,圣忠华1 ,邵高能1 ,唐绍清1 ,魏祥进1,* ,胡培松1, *

doi:10.3969/j.issn.1001-7216.2014.04.005

中国水稻科学 >

2014 , Vol. 28 >Issue 4: 367 - 376

DOI: https://doi.org/10.3969/j.issn.1001-7216.2014.04.005

研究报告

水稻类病斑突变体lmm4的鉴定及其基因定位

展开

¹中国水稻研究所/农业部水稻生物学与遗传育种重点实验室/水稻生物学国家重点实验室，杭州 310006；²杭州师范大学生命与环境科学学院，杭州 310036；

收稿日期: 2014-01-16

修回日期: 2014-03-01

网络出版日期: 2014-07-10

基金资助

国家自然科学基金资助项目（31201193, 31201195）；国家863计划资助项目（2011AA10A101, 2012AA101101）。

收起

Identification and Gene Mapping of a Lesion Mimic Mutant lmm4 in Rice

Expand

¹ State Key Laboratory of Rice Biology, Key Laboratory of Rice Biology and Breeding of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China; ²College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China;

Received date: 2014-01-16

Revised date: 2014-03-01

Online published: 2014-07-10

Fold

摘要

在EMS诱变的日本晴突变体库中，筛选到一个类病斑突变体。该突变体类病斑出现于分蘖始期，而后慢慢扩散至叶鞘、茎秆，最后至整个植株，属于扩散型类病斑突变体。相比野生型，突变体的lmm4 的株高明显降低，有效穗数减少，穗长缩短，结实率、每穗实粒数、千粒重均显著降低。遮光处理实验表明，突变体lmm4 类病斑的产生受自然光诱导。突变体lmm4 的光合色素含量和净光合速率明显低于野生型。电子显微镜观察发现突变体类病斑部位叶肉细胞中的叶绿体结构发育异常。台盼蓝染色实验表明突变体类病斑部有大量的死亡细胞。二氨基联苯胺染色实验表明类病斑部位有过氧化氢沉积。稻瘟病抗性鉴定结果表明，与野生型相比，突变体lmm4 对C7和G1等稻瘟病菌生理小种的抗性明显增强。实时PCR分析证明在突变体lmm4 中防卫反应相关基因POC1、PAL、PBZ1、PR1的表达量显著提高。遗传分析表明突变体表型符合单隐性核基因控制的遗传规律。lmm4 定位在第5染色体着丝粒附近Ind4和Ind6之间大约235 kb的区域内。

关键词： 类病斑突变体; 稻瘟病抗性; 基因定位; 水稻

本文引用格式

邱结华1 ,马宁1, 2,蒋汉伟1, 2 ,圣忠华1 ,邵高能1 ,唐绍清1 ,魏祥进1,* ,胡培松1, * . 水稻类病斑突变体lmm4的鉴定及其基因定位[J]. 中国水稻科学, 2014 , 28(4) : 367 -376 . DOI: 10.3969/j.issn.1001-7216.2014.04.005

Abstract

The lesion mimic mutant lmm4 was identified from an EMSinduced Nipponbare mutant library. Brown lesions were first observed on the tip of lmm4 leaves at the initial tillering stage, and spreaded gradually downward to the whole leaf blade and even the leaf sheath. Compared with the wild type, the chlorophyll and the net photosynthetic rate of lmm4 were significantly reduced. The plant height, panicle length, seed setting rate, grain weight were also decreased significantly. The shading assay showed that the lesions on lmm4 leaves were induced by natural light. The histological staining assay indicated that the lesions on lmm4 leaves were caused by programmed cell death and hydrogen peroxide accumulated in lmm4 leaves. The lmm4 displayed higher resistance to blast race C7 and G1 when compared with the wild type. In addition, pathogenesisrelated gene POC1, PAL,PBZ1, PR1 were found to be upregulated in lmm4. Genetic analysis suggested that the phenotype of lmm4 was controlled by a single recessive nuclear locus. Based a mapbased cloning strategy, the lmm4 was narrowed down to a 235 kb region between Ind4 and Ind6 near the centromere of chromosome 5.

Key words： lesion mimic mutant; blast resistance; gene mapping; rice

参考文献

\[1\]Hu G, Richter T E, Hulbert S H, et al. Disease lesion mimicry caused by mutations in the rust resistance gene rp1. Plant Cell, 1996, 8(8):13671376.

\[2\]Dangl J L, Dietrich R A, Richberg M H. Death don’t have no mercy: Cell death programs in plantmicrobe interactions.Plant Cell, 1996, 8(10): 17931807.

\[3\]陈析丰, 金杨, 马伯军. 水稻类病变突变体及抗病性的研究进展. 植物病理学报, 2011, 41(1): 19.

\[4\]王建军, 朱旭东, 王林友, 等. 水稻类病变突变体lrd40的抗病性与细胞学分析. 中国水稻科学, 2005, 19(2): 111116.

\[5\]Qiao Y L, Jiang W Z, Lee J H, et al. SPL28 encodes a clathrinassociated adaptor protein complex 1, medium subunit μ1 (AP1M1) and is responsible for spotted leaf and early senescence in rice (Oryza sativa). New Phytol, 2010, 185(1):258274.

\[6\]黄奇娜, 杨杨, 施勇烽, 等. 水稻斑点叶变异研究进展. 中国水稻科学, 2010, 24(2):108115.

\[7\]Yamanouchi U, Yano M, Lin H, et al. A rice spotted leaf gene,Spl7, encodes a heat stress transcription factor protein. Proc Natl Acad Sci, 2002, 99(11):75307535.

\[8\]Zeng L R, Qu S H, Bordeos A, et al. Spotted leaf 11, a negative regulator of plant cell death and defense, encodes a Ubox/armadillo repeat protein endowed with E3 ubiquitin ligase activity. Plant Cell, 2004, 16(10):27952808.

\[9\]Wang L J, Pei Z Y, Tian Y C, et al. OsLSD1, a rice zinc finger protein, regulates programmed cell death and callus differentiation. Mol PlantMicrobe Interact, 2005, 18(5):375384.

\[10\]Yuan Y X, Zhong S H, Li Q, et al. Functional analysis of rice NPR1like genes reveals that OsNPR1/NH1 is the rice orthologue conferring disease resistance with enhanced herbivore susceptibility. Plant Biotechnol J, 2007, 5(2):313324.

\[11\]Kim J A, Cho K, Singh R, et al. Rice OsACDR1 (Oryza sativa accelerated cell death and resistance 1 ) is a potential positive regulator of fungal disease resistance. Mol Cell, 2009, 28(5):431439.

\[12\]Mori M, Tomita C, Sugimoto K, et al. Isolation and molecular characterization of a Spotted leaf 18 mutant by modified activationtagging in rice. Plant Mol Biol, 2007, 63(6):847860.

\[13\]Takahashi A, Agrawal G K, Yamazaki M, et al. Rice Pti1a negatively regulates RAR1dependent defense responses. Plant Cell, 2007, 19(9):29402951.

\[14\]Jiang C J, Shimono M, Maeda S, et al. Suppression of the rice fattyacid desaturase gene OsSSI2 enhances resistance to blast and leaf blight diseases in rice. Mol PlantMicrobe Interact, 2009, 22(7):820829.

\[15\]Lichtenthaler H K. Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes. Methods Enzymol, 1987, 148:350382.

\[16\]吕典华, 宗学凤, 王三根, 等. 两个水稻叶色突变体的光合特性研究. 作物学报, 2009, 35(12):23042308.

\[17\]Yin Z C, Chen J, Zeng L R, et al. Characterizing rice lesion mimic mutants and identifying a mutant with broadspectrum resistance to rice blast and bacterial blight. Mol PlantMicrobe Interact, 2000, 13(8):869876.

\[18\]ThordalChristensen H, Zhang Z, Wei Y, et al. Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barleypowdery mildew interaction. Plant J, 1997, 11(6):11871194.

\[19\]Matsumoto T, Wu J, Kanamori H, et al. The mapbased sequence of the rice genome. Nature, 2005, 436(7052):793800.

\[20\]Edwards K, Johnstone C, and Thompson C. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis.Nucl Acids Res, 1991, 19(6):1349.

\[21\]Jiao B B, Wang J J, Zhu X D, et al. A novel protein RLS1 with NBARM domains is involved in chloroplast degradation during leaf senescence in rice. Mol Plant, 2012, 5(1):205217.

\[22\]Feng B H, Yang Y, Shi Y F, et al. Characterization and genetic analysis of a novel rice spottedleaf mutant HM47 with broadspectrum resistance to Xanthomonas oryzae pv. oryzae. J Integr Plant Biol, 2013, 55(5):473483.

\[23\]Ishikawa A, Okamoto H, Iwasaki Y, et al. A deficiency of coproporphyrinogen III oxidase causes lesion formation in Arabidopsis.Plant J, 2001, 27(2):8999.

\[24\]Noutoshi Y, Ito T, Seki M, et al. A single amino acid insertion in the WRKY domain of the Arabidopsis TIRNBSLRRWRKYtype disease resistance protein SLH1 (sensitive to low humidity 1) causes activation of defense responses and hypersensitive cell death. Plant J, 2005, 43(6):873888.

\[25\]Wu C, Bordeos A, Madamba M R S, et al. Rice lesion mimic mutants with enhanced resistance to diseases.Mol Genet Genom, 2008, 279(6):605619.

Options

文章导航

模态框（Modal）标题

摘要

本文引用格式

Abstract

参考文献