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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (02): 204-213.doi: 10.3724/SP.J.1006.2020.92020

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

Phenotype characterization and gene mapping of the semi-outcurved leaf mutant sol1 in rice (Oryza sativa L.)

XIE Yuan-Hua,LI Feng-Fei,MA Xiao-Hui,TAN Jia,XIA Sai-Sai,SANG Xian-Chun,YANG Zheng-Lin,LING Ying-Hua()   

  1. Rice Research Institute of Southwest University / Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Chongqing 400715, China
  • Received:2019-04-09 Accepted:2019-08-09 Online:2020-02-12 Published:2019-09-03
  • Contact: Ying-Hua LING E-mail:lingyh003@126.com
  • Supported by:
    This study was supported by the National Key R&D Program of China(2017YFD0100201);the Special Project on Science and Technology Innovation for Social Undertakings and Livelihood Security in Chongqing(cstc2016shms-ztcx80012);the Fundamental Research Funds for the Central Universities(XDJK2016A013)

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Abstract:

Moderate curling of leaves helps to improve the photosynthetic efficiency, and increase the overall effective accumulation of photosynthate. We identified a stable mutant from the library constructed by treating indica maintainer line Xinong 1B with ethyl methane sulfonate (EMS). Leaves of the mutant gradually curled outwards into a semi-coiled shape, accompanied by semi-dwarfing of the culm and leaf drooping from the 10-leaf stage, the mutant was temporarily named as semi-outcurved leaf 1 (sol1). The calculated leaf curl index of sol1 was 30%, statistically higher than that of wild type (WT, P < 0.01). Panicle length and internode length of Top 1 (the 1st leaf from top), Top 2 (the 2nd leaf from top), Top 3 (the 3rd leaf from top), and Top 4 (the 4th leaf from top) were very significantly decreased. The leaf angles of Top 1, Top 2, and Top 3 were significantly or very significantly increased. The number of effective panicles, 1000-grain weight, filled grain number per panicle, and seed setting rate of the mutant sol1 were significantly or very significantly decreased, while primary branch number increased by 11.3%, statistically higher than that of WT (P < 0.05). The transpiration rate, intercellular CO2 concentration and stomatal conductance of sol1 were significantly higher than those of wild type. The results of paraffin section showed that the bulliform cells of the Top1 of sol1 exhibited smaller, but their number increased significantly, and the volume of epidermal cells increased slightly. Genetic analysis indicated that sol1 was controlled by a recessive nuclear gene. SOL1 was precisely located in the physical distance of 165 kb between the markers of JY6-3 and JY6-10 on the short arm of chromosome 6, with 15 annotated genes. The results of qRT-PCR showed that RL14, Roc5, and REL1 related to bulliform cells were up- regulated in mutant sol1, while NRL, BRD1, OsHox32, ADL1, LC2 down-regulated. The results of the present study provide a basis for SOL1 cloning, and functional dissection as well.

Key words: rice (Oryza sativa L.), semi-outcurved leaf mutant sol1, phenotype characterization, gene mapping, bulliform cell

Fig. 1

Phenotypic identification of the wild type(WT) and the sol1 mutant A, B, C, D: plant phenotypes of WT and sol1 at the 5-6 leaf stage, the tillering stage and the mature stage, respectively; E and F: incompletely extracted leaf base of WT and sol1, respectively; G: bases of the abaxial top 1 of WT and sol1; H: tips of the adaxial top 1 of WT and sol1; I: leaf curl indexes of WT and sol1 at jointing stage; J: lengths of the spike and internodes of WT and sol1; K: comparison of the internodes of WT and sol1 at mature period; L: leaf angles of WT and sol1 at the shooting period. Bar = 10 cm, in A, B, C, D, J diagrams; Bar = 2 cm, in E, F, G, H diagrams. Top1: the 1st leaf from the top; Top2: the 2nd leaf from the top; Top3: the 3rd leaf from the top. * means significant difference at 0.01 ≤ P < 0.05 by t-test; ** means significant difference at P < 0.01 by t-test."

Fig. 2

Agronomic characters of the wild type (WT) and the sol1 mutant A, B: length and width of tops 1, 2, 3 at booting stage (cm); C: numbers of effective panicles; D: primary branches; E: secondary branches; F: 1000-grain weights (g); G: filled grain numbers per panicle; H: Seed setting rate (%); Top 1: the 1st leaf from the top; Top 2: the 2nd leaf from the top; Top 3: the 3rd leaf from the top. * means significant difference at 0.01 ≤ P < 0.05 by t-test; ** means significant difference at P < 0.01 by t-test."

Fig. 3

Analysis of photosynthetic efficiency of blade at maturation period A: net photosynthetic rate (Pn) of tops 1, 2, 3; B: transpiration rate (Tr) of tops 1, 2, 3; C: intercellular CO2 concentration (Ci) of tops 1, 2, 3; D: stomatal conductance (Gs) of the tip and base of tops 1, 2, 3. Top 1: the 1st leaf from the top; Top 2: the 2nd leaf from the top; Top 3: the 3rd leaf from the top. * significant difference at P < 0.05; ** significant difference at P < 0.01."

Fig. 4

Paraffin-cut section analysis of cross section of leaf A, B: cross cutting diagrams of top 1 of WT and sol1 at the heading stage, respectively, Bar = 1 mm; C, D: magnified diagrams of a red box in picture A and B, respectively; E, F: magnified diagrams of a yellow box in picture A and B, respectively, Bar = 0.1 mm; G: number of bulliform cells between two small vein; H: area of bulliform cells between two small vein. BC: bulliform cell. * means significant difference at 0.01 ≤ P < 0.05 by t-test."

Fig. 5

Location of candidate gene SOL1"

Table 1

Newly developed linkage primers on chromosome 6"

引物名称
Primer		 正向引物
Forward primer (5′-3′)		 反向引物
Reverse primer (5′-3′)
RM204 CTAGCTAGCCATGCTCTCGTACC CTGTGACTGACTTGGTCATAGGG
ZTQ54 AAATAGATAGTAACGGCTCTAACATG CTTGCACAACTGCTGTTCATG
ZZC6-6 TCCATTGCTCCCAAACACTC CGTGGAACCAAATACAGCCT
JY6-7 GTGAGTGCTAAGGATGATGTGTTG TGCAGGAAGGCATTGTATCC
JY6-4 GCAGCAAAGATCCATACGAGGT AATCTTGGCGGCTCGTGTAG
JY6-3 CGAACACATGCCACACACACA GGGCTAGACAAACAATAATGTGC
JY6-10 TGGCAGCGGTGGTGGTATAT CAATGAGAAGTTGGTGGTGTTCT

Table 2

Candidate gene annotation in the location interval"

基因命名
Gene nomenclature		 基因注释
Gene annotation
LOC_Os06g10540 表达蛋白 Expressed protein
LOC_Os06g10550 反转录转座子 Retrotransposon protein, putative, unclassified, expressed
LOC_Os06g10560 叶片衰老相关蛋白 Leaf senescence related protein, putative, expressed
LOC_Os06g10570 表达蛋白 Expressed protein
LOC_Os06g10580 环核苷酸门控离子通道 Cyclic nucleotide-gated ion channel, putative, expressed
LOC_Os06g10590 表达蛋白 Expressed protein
LOC_Os06g10600 同源异型结构域和START结构域 Homeobox and START domains containing protein, putative, expressed
LOC_Os06g10610 表达蛋白 Expressed protein
LOC_Os06g10620 转录延伸因子复合物SPT5 Transcription elongation factor SPT5 homolog, putative, expressed
LOC_Os06g10650 蛋白质酪氨酸磷酸酶家族 Tyrosine phosphatase family protein, putative, expressed
LOC_Os06g10660 锚定蛋白前体LysM Domain-containing GPI-anchored protein 1 precursor, putative, expressed LysM-GPI
LOC_Os06g10670 天冬氨酸蛋白酶的猪笼草蛋白前体 Aspartic proteinase nepenthesin-1 precursor, putative, expressed
LOC_Os06g10680 表达蛋白 Expressed protein
LOC_Os06g10710 表达蛋白 Expressed protein
LOC_Os06g10720 表达蛋白 Expressed protein

Fig. 6

qRT-PCR of genes related to curled leaf and dwarf internode A: some genes in bulliform cell development-related rolling leaf; B: genes about some abaxially curled leaf."

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