Characterisation and Gene Mapping of a Brittle Culm Mutant bc-s1 in Rice

doi:10.11983/CBB15048

Abstract

Abstract: The mechanical strength of rice stem is an important agronomic trait that affects crop lodging. In this study, a brittle culm mutant, bc-s1, was identified from the rice mutant library. The mutant showed inherited traits of brittleness from bolting to mature stage. Meanwhile, the leaf, leaf sheath, culm, node and seed shell of the mutant showed brittleness during the mature stage. Compared with breaking and elongation force in the wild type, that in bc-s1 culms decreased 31.1% and 67.2%, respectively. Besides showing irregular shape of sclerenchyma cells, in bc-s1, cellulose and lignin content in culm decreased 24.97% and increased 38.82%, respectively. The BC-S1 gene was mapped on chromosome 9 between InDel markers Ha6 and Ha9 within 101 kb physical distance. According to sequencing results for bc-s1, a candidate open reading frame (Os09g25490/OsCesA9), which encodes a cellulose synthase catalytic subunit (CesA), showed a point mutation at the 28^th base of first extron, changing GGG to AGG and causing the substitution at the 10th amino acid residue from glycine to arginine. Therefore, bc-s1 is a new allele of OsCesA9 and provides a new material for uncovering the biological function of OsCesA9 in rice cell-wall biosynthesis.

Key words: bc-s1, brittle culm, secondary wall, cellulose synthase, rice

Zhenming Jin, Baozhe Ping, Haojun Shen, Huaiqing Du, Ruiqian Li, Lu Zhu, Dabing Zhang, Zheng Yuan. Characterisation and Gene Mapping of a Brittle Culm Mutant bc-s1 in Rice[J]. Chinese Bulletin of Botany, 2016, 51(2): 167-174.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB15048

https://www.chinbullbotany.com/EN/Y2016/V51/I2/167

Figures/Tables 5

Table 1 DNA markers used for positional cloning

Marker	Forward primer (5'-3')	Reverse primer (5'-3')	Type
Chr09_04	GGTAATGTCACAACTCAAAAAGC	CAAGATTGTAAACCCTGTCTATTG	InDel
Ha1	GCGAACCGATAAAACTGCTC	AGAGGTGTATCAAAGCAATCGAG	InDel
Ha4	TCACCTCTCAACTTAATCGA	AGTCCATCAAGCCATGATGC	InDel
Ha6	AGTTCGTCCGGTTTTGATCG	GTAGAATAAGCGAAACAGCA	InDel
Ha9	TCGACCATCAGCGATTTGAC	TTTTCCATGCGCGGTGTTTG	InDel
Chr09_05	GAATTTGAGTGGGTTTATACTAGC	GTTAACTTAGGCTATTTTGGCTTC	InDel

Figure 1 The phenotype and mechanical strength of bc-s1 mutant and wild-type 9522 in Oryza sativa (A) The whole plant phenotype of bc-s1 mutant and wild-type 9522 in the heading stage (Bar=20 cm); (B) An easily broken culm of bc-s1 compared with a wild-type 9522 culm in the mature stage; (C) An easily broken flag leaf of bc-s1 compared with a wild-type 9522 flag leaf in the mature stage; (D) The elongation force of the second upper internode; (E) The breaking force of the second upper internode. * and ** represent t-test at P<0.05 and P<0.01, respectively.

Figure 2 The cross-section of wild-type 9522 and bc-s1 mutant culms in Oryza sativa under a scanning electron microscope (A), (B) The primary wall of a wild-type 9522 culm; (C), (D) The secondary wall of a wild-type 9522 culm; (E), (F) The primary wall of a bc-s1 mutant culm; (G), (H) The secondary wall of a bc-s1 mutant culm; (A), (C), (E), (G) Bar=100 μm; (B), (D), (F), (H) Bar=5 μm

Figure 3 Tissue section staining and components determination in the culm cell wall of wild-type 9522 and bc-s1 mutant in Oryza sativa (A), (B) The cross section of the second upper internodes of wild-type 9522 (A) and bc-s1 mutant (B) were stained with phloroglucinol; (C), (D) The cross section of the second upper internodes of wild-type 9522 (C) and bc-s1 mutant (D) were stained with Calcofluor White; (E) The comparison of cell-wall components between bc-s1 mutant and wild-type 9522 culm. * and ** represent t-test at P<0.05 and P<0.01, respectively.

Figure 4 Map-based cloning of BC-S1 The BC-S1 gene (Os09g25490/OsCesA9), which encodes a cellulose synthase catalytic subunit (CesA), was mapped on BAC AP005579, chromosome 9. According to sequence comparison result, the BC-S1 showed a point mutation at the 28th base of first extron (changing GGG to AGG)

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