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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (1): 46-54.doi: 10.3724/SP.J.1006.2019.82022

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2018-09-18 Published:2018-09-18
  • Contact: Ying-Hua LING E-mail:lingyh003@126.com
  • 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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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

Fig. 1

Phenotypes of the wild type (WT) and the lmps1 mutant A: plants of the wild type and the lmps1 mutant at tillering stage; B: leaves of the wild type and the lmps1 mutant at tillering stage; C: plants of the wild type and the lmps1 mutant at mature period; D: leaves of the wild type and the lmps1 mutant at mature period; 1: the flag leaves; 2: the second leaves; 3: the third leaves; bar = 5 cm."

Table 1

Phenotypic difference between the wild type and the lmps1 mutant"

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

Fig. 2

Effects of shading on the wild type (WT) and the lmps1 mutant leaves A: leaf of the wild type; B: leaf of the lmps1 mutant; C: leaf of the lmps1 mutant after one week shading; D: leaf of the lmps1 mutant illuminated for one week after shading; bar = 6 cm."

Fig. 3

Photosynthetic pigments contents of the wild type (WT) and the lmps1 mutant at booting stage A-D: photosynthetic pigments contents of the flag leaves, the second leaves, the third leaves and the fourth leaves respectively in the wild type and the lmps1 mutant at booting stage; E: photosynthetic pigments contents of the wild type; F: photosynthetic pigments contents of the lmps1 mutant. The column shows the mean value and the error line shows the standard deviation; ** means significant difference at P < 0.01 by t-test; in Figs. E and F, bars superscripted by different letter are significantly different at P < 0.05."

Fig. 4

Photosynthetic characteristics of the wild type (WT) and the lmps1 mutant at booting stage A: net photosynthetic rate of the wild type and the lmps1 mutant; B: stomatal conductance of the wild type and the lmps1 mutant; C: intercellular CO2 concentration of the wild type and the lmps1 mutant; D: transpiration rate of the wild type and the lmps1 mutant. The column shows the mean value and the error line shows the standard deviation; ** means significant difference at P < 0.01 by t-test, * means significant difference at P < 0.05 by t-test."

Fig. 5

Ultrastructure of mesophyll cells in the wild type (WT) and the lmps1 mutant at booting stage A: tip of the fourth leaves of the wild type; B, C, D: ultrastructure of the blade tip of the wild type; E: tip of the fourth leaves of the lmps1 mutant; F, G, H: ultrastructure of the blade tip of the lmps1 mutant; I: middle part of the fourth leaves of the wild type; J, K, L: ultrastructure of the middle part of blade of the wild type; M: middle part of the fourth leaves of the lmps1 mutant; N, O, P: ultrastructure of the middle part of blade of the lmps1 mutant. Chl: chloroplast; Os: osmiophilic granule; TM: thylakoid membranes; N: ribosome. Bars: 2 μm (A, D), 1 μm (B, E), 200 nm (C, F)."

Fig. 6

Antioxidant enzyme activity and the H2O2 content in the wild type (WT) and the lmps1 mutant at the booting stage A-C: the enzyme activity of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in functional leaves of the wild type and the lmps1 mutant; D: H2O2 content of different functional leaves of the wild type and the lmps1 mutant. The column shows the mean value and the error line shows the standard deviation; ** means significant difference at P < 0.01 by t-test, * means significant difference at P < 0.05 by t-test."

Fig. 7

Molecular mapping of LMPS1 on chromosome 7 A: QTL mapping of LMPS1; B: fine mapping of LMPS1."

Fig. 8

Relative expression level of genes in the wild type (WT) and the lmps1 mutant A: relative expression level of annotated genes within the physical fine mapping region of LMPS1; B: relative expression level of genes related to disease resistance. The column shows the mean value and the error line shows the standard deviation; * refers to the significant difference at P < 0.05; ** denotes significant difference at P < 0.01."

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