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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (5): 1426-1431.doi: 10.3724/SP.J.1006.2023.22025

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Identification and gene mapping of brittle culm mutant bc21 in rice

DAI Wen-Hui1,2(), ZHU Qi1,2, ZHANG Xiao-Fang1,2, LYU Shen-Yang1, XIANG Xian-Bo3, MA Tao1, CHEN Yu-Jie1, ZHU Shi-Hua1, DING Wo-Na1,*()   

  1. 1College of Science and Technology, Ningbo University, Ningbo 315212, Zhejiang, China
    2School of Marine Science, Ningbo University, Ningbo 315211, Zhejiang, China
    3Pingyang Administration for Market Regulation, Wenzhou 325400, Zhejiang, China
  • Received:2022-04-27 Accepted:2022-07-22 Online:2023-05-12 Published:2022-08-12
  • Contact: *E-mail: dwn@zju.edu.cn
  • Supported by:
    National Natural Science Foundation of China(32071981);Key Project of Ningbo Natural Science Foundation(202003N4016)

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Abstract:

A brittle culm mutant bc21 was obtained by mutagenesis of indica rice Kasalath with ethyl methanesulfonate (EMS). Phenotypic analysis revealed that the mutant displayed both brittle culm and leaf phenotypes, which began to manifest at seedling stage and were most pronounced at mature stage. The mechanical strength analysis showed that the breaking resistance and tensile force of bc21 stems decreased significantly. Resin sections and scanning electron microscope observation showed that the sclerenchyma cells of bc21 culms had increased voids and thinner cell walls. Compared with the wild type, the cell wall component of stems revealed that the cellulose content of bc21 decreased by 36.60%, and the hemicellulose and lignin contents increased by 23.08% and 26.06%, respectively. Genetic analysis indicated that the brittle trait of bc21 was controlled by a single recessive gene. Using SSR markers and self-designed STS markers, BC21 was located in a 52.9 kb region between the markers STS2 and STS3 on chromosome 6, and there was no previously reported rice brittleness-related gene within this region, indicating that BC21 might be a new brittle culm gene in rice. This study will provide material support for further dissection of the regulation mechanism of mechanical strength of rice stems.

Key words: rice, brittle culm mutant, cell wall, cellulose, gene mapping

Fig. 1

Broken phenotypes of wild type (WT) and mutant bc21 stems and leaves at maturity stage A: the broken phenotypes of stems; B: the broken phenotypes of leaves. WT: Kasalath; bc21: brittle culm 21."

Fig. 2

Breaking resistance (A) and tensile force (B) of the second internodes of wild type (WT) and mutant bc21 * represents significant difference at the 0.05 probability level; ** represents significant differences at the 0.01 probability level. WT: Kasalath; bc21: brittle culm 21."

Fig. 3

Cross section of wild type and mutant bc21 in nodes A: the observation on resin slices of wild type stem cross section (bar: 100 μm); B: the observation on resin slices of bc21 stem cross section (bar: 100 μm); C: the scanning electron microscope observation of wild type stem cross section (bar: 20 μm); D: the scanning electron microscope observation of bc21 stem cross section (bar: 20 μm)."

Fig. 4

Cellulose, hemicellulose, and lignin content in wild type (WT) and mutant bc21 * represents significant differences at the 0.05 probability level. WT: Kasalath; bc21: brittle culm 21."

Table 1

SSR and STS molecular markers for mapping"

标记
Marker name		 正向引物
Forward primer (5'-3')		 反向引物
Reverse primer (5'-3')
RM6734 TGAGCAGTCTGCAGATGACC GCTTGGACTTGGAGTCTTGG
STS1 ACCAGGCTGAATGTATAGAT TTAGGCACATAAACCAAG
STS2 AAAATTGTAGTGGGTTGGT TACAGAGAAAAAGATTGAAGC
STS3 CAGTGATTCGTTTGAAAT CCCTGTTGTTTGTATGAC
RM253 TCCTTCAAGAGTGCAAAACC GCATTGTCATGTCGAAGCC

Fig. 5

Fine mapping of BC21 on rice chromosome 6"

Table 2

Gene names and their putative functions in the target interval"

基因名称Gene name 推测功能Putative function
LOC_Os06g09570 核仁GTP结合蛋白1 Nucleolar GTP-binding protein 1
LOC_Os06g09580 转座子蛋白 Transposon protein, putative
LOC_Os06g09600 表达蛋白 Expressed protein
LOC_Os06g09610 过氧化物还原酶 Peroxiredoxin
LOC_Os06g09620 表达蛋白 Expressed protein
LOC_Os06g09630 3-氧化酰基合酶 3-oxoacyl-synthase
LOC_Os06g09640 反转录转座子蛋白 Retrotransposon protein
LOC_Os06g09650 反转录转座子蛋白 Retrotransposon protein
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