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作物学报 ›› 2019, Vol. 45 ›› Issue (1): 46-54.doi: 10.3724/SP.J.1006.2019.82022

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻类病斑早衰突变体lmps1的表型鉴定与基因定位

夏赛赛,崔玉,李凤菲,谭佳,谢园华,桑贤春,凌英华()   

  1. 西南大学水稻研究所 / 转基因植物与安全控制重庆市重点实验室, 重庆 400715
  • 收稿日期:2018-04-25 接受日期:2018-08-20 出版日期:2018-09-18 网络出版日期:2018-09-18
  • 通讯作者: 凌英华
  • 基金资助:
    本研究由重庆市社会事业与民生保障科技创新专项项目(cstc2016shms-ztcx80012);中央高校基本科研业务费专项资金资助(XDJK2016A013)

Phenotypic characterizing and gene mapping of a lesion mimic and premature senescence 1 (lmps1) mutant in rice (Oryza sativa L.)

Sai-Sai XIA,Yu CUI,Feng-Fei LI,Jia TAN,Yuan-Hua XIE,Xian-Chun SANG,Ying-Hua LING()   

  1. Rice Research Institute of Southwest University / Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Chongqing 400715, China
  • Received:2018-04-25 Accepted:2018-08-20 Published:2018-09-18 Published online:2018-09-18
  • Contact: Ying-Hua LING
  • Supported by:
    This study was supported by the project of Chongqing Science and Technology Commission Grants(cstc2016shms-ztcx80012);Fundamental Research Funds for the Central Universities(XDJK2016A013)

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摘要：

经甲基磺酸乙酯(EMS)诱变优良籼型水稻恢复系缙恢10号, 获得一个稳定遗传的水稻类病斑早衰突变体lmps1 (lesion mimic and premature senescence 1)。该突变体苗期表型正常, 分蘖早期出现褐色类病斑, 且斑点数目随植株生长而增多, 孕穗期叶片开始萎黄衰老。与野生型相比, 突变体lmps1的每穗总粒数下降8% (P<0.05), 株高、穗长、有效穗数、每穗实粒数、结实率以及千粒重分别下降14.3%、24.3%、27.2%、50%、45.7%与14.5%, 差异均达极显著水平(P<0.01)。遮光处理表明, 突变体lmps1的类病斑性状受光照诱导。孕穗期叶片光合色素含量下降且光合效率降低, H2O2含量增加, 抗氧化酶SOD和CAT的活性显著降低。透射电镜观察结果显示, 突变体lmps1叶肉细胞中叶绿体数目减少, 叶绿体的类囊体片层结构损伤降解。qRT-PCR结果显示, 突变体lmps1中防卫反应相关基因除POX22.3表达量降低外, POC1、PAL、PBZ1、PR1、NPR1、PR5表达量均极显著高于野生型。遗传分析表明突变体lmps1的类病斑早衰性状受1对隐性核基因控制, 利用西农1A与突变体lmps1杂交所得F2群体中的突变株, 将目标基因定位于第7染色体长臂端粒附近约167.3 kb的物理区段内。

关键词: 水稻(Oryza sativa L.), 类病斑早衰突变体 (lmps1), 表型鉴定, 基因定位

Abstract:

An elite rice indica restorer line, Jinhui 10 was chemically mutated with ethyl methane sulfonate (EMS). A novel and stable mutant, lesion mimic and premature senescence 1 (lmps1) was identified from the descendant library. The growth and development of lmps1 plant was normal at seedling stage, but its leaves began to exhibit brown spots at the beginning of tillering stage, and the number of the spots increased as the plant growing up. When mutant plant entered booting stage, leaves started to wilt and became yellowish, then exhibited premature senescence. Compared with wild type, grains per panicle of lmps1 decreased by 8%, significantly lower than that of wild type (P < 0.05). Besides, plant height, panicle length, effective panicle number, filled grain number per panicle, seed setting rate, and 1000-grain weight of lmps1 correspondingly decreasedly 14.3%, 24.3%, 27.2%, 50.0%, 45.7%, and 14.5%, statistically lower than those of wild type (P < 0.01). The shading treatment suggested that the mutant performance of lmps1 was light induced. Within the cells of lmps1, both of photosynthetic pigment content and photosynthetic efficiency decreased. At the same time, H2O2 content increased, and activities of protective enzymes, SOD and CAT, reduced in lmps1. The observation by transmission electronic microscope (TEM) demonstrated that chloroplast number reduced in lmps1, and the chloroplast thylakoid lamellar structure seriously damaged and degraded. As a result of qRT-PCR, the expression levels of POC1, PAL, PBZ1, PR1, NPR1, and PR5 were higher in lmps1 than in wild type, while that of POX22.3 was lower than that of wild type. The genetic analysis indicated that the lesion mimic and premature senescence of lmps1 was under the control of a recessive nuclear gene. Mapping result via F2 population derived from the cross between Xinong 1A and lmps1 showed that, the target gene was located in a 167.3 kb physical interval near the telomere of the long arm of chromosome 7 in rice (Oryza sativa L.).

Key words: rice (Oryza sativa L.), lesion mimic and premature senescence 1 (lmps1), phenotyping, gene mapping

图1

野生型(WT)与突变体lmps1的表型 A: 分蘖期WT与lmps1植株; B: 分蘖期WT与lmps1的叶片; C: 成熟期WT与lmps1植株; D: 成熟期WT与lmps1的叶片; 1: 倒一叶; 2: 倒二叶; 3: 倒三叶; 标尺=5 cm。"

表1

野生型与突变体lmps1之间的表型差异"

性状
Trait		 野生型
Wild type		 突变体
lmps1		 lmps1较WT减少率
Reduction in lmps1 than in WT (%)
株高 Plant height (cm) 108.50±2.29 93.00±2.00** 14.3
穗长 Panicle length (cm) 26.15±0.55 19.80±1.03** 24.3
有效穗数 Effective panicle 9.62±1.31 7.00±1.07** 27.2
每穗总粒数 Grain number per panicle 161.34±6.78 148.42±13.40* 8.0
每穗实粒数 Filled grain number per panicle 133.48±6.36 66.27±7.66** 50.0
结实率 Seed setting rate (%) 82.75±2.51 44.97±5.95** 45.7
千粒重 1000-grain weight (g) 25.74±0.44 22.01±0.43** 14.5

图2

遮光对野生型(WT)和突变体lmps1叶片的影响 A: WT叶片; B: lmps1叶片; C: lmps1遮光1周后; D: lmps1遮光处理后复光1周后; 标尺=6 cm。"

图3

野生型(WT)与突变体lmps1孕穗期光合色素含量 A~D: 孕穗期WT与lmps1的倒一叶(A)、倒二叶(B)、倒三叶(C)和倒四叶(D)光合色素含量; E: WT光合色素含量; F: lmps1光合色素含量。条柱表示平均值, 误差线表示标准差; **表示在0.01水平上差异显著; E, F图中标的不同字母的柱值在0.05水平上差异显著。"

图4

孕穗期野生型(WT)与突变体lmps1功能叶的光合特性 A: WT和lmps1的净光合速率; B: WT和lmps1的气孔导度; C: WT和lmps1的细胞间CO2浓度; D: WT和lmps1的蒸腾速率。条柱表示平均值, 误差线表示标准差; **表示在0.01水平上差异显著, *表示在0.05水平上差异显著。"

图5

孕穗期野生型(WT)和突变体lmps1的叶肉细胞超微结构 A: WT倒四叶尖; B、C、D: WT叶尖细胞超微结构; E: lmps1倒四叶尖; F、G、H: lmps1叶尖细胞超微结构; I: WT倒四叶中部; J、K、L: WT叶中部细胞超微结构; M: lmps1倒四叶中部; N、O、P: lmps1叶中部细胞超微结构; Chl: 叶绿体; Os: 嗜锇小体; TM: 类囊体膜; N: 细胞核。标尺: 2 μm (A, D), 1 μm (B, E), 200 nm (C, F)。"

图6

孕穗期野生型(WT)和突变体lmps1的抗氧化酶活性及H2O2含量 A~C: WT和lmps1的过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)活性; D: WT和lmps1各功能叶过氧化氢(H2O2)含量。条柱表示平均值, 误差线表示标准差; **表示在0.01水平上差异显著, *表示在0.05水平上差异显著。"

图7

LMPS1基因在第7染色体的分子定位 A: LMPS1基于F2群体的初定位; B: LMPS1的精细定位。"

图8

野生型(WT)和突变体lmps1相关基因的表达量 A: LMPS1精细定位物理区间内部分基因的表达量; B: 抗病相关基因相对表达量。条柱表示平均值, 误差线表示标准差; *表示0.05水平上差异显著; **表示在0.01水平上差异显著。"

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