Comparative analysis of different types of callus in moso bamboo

doi:10.12302/j.issn.1000-2006.202203062

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (6): 141-149.doi: 10.12302/j.issn.1000-2006.202203062

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Comparative analysis of different types of callus in moso bamboo

HUANG Biyun¹^,²(), ZHUO Renying¹, QIAO Guirong¹^,^*()

1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry,Hangzhou 311400, China
2. College of Forestry and Grassland,Nanjing Forestry University, Nanjing 210037,China

Received:2022-03-28 Revised:2022-06-19 Online:2023-11-30 Published:2023-11-23

Abstract

Abstract:

【Objective】 The proliferation ability and differentiation potential of different types of moso bamboo callus were compared with reference to three aspects: cell morphology, cell cycle phase distribution ratio and gene expression. The results provided a basis for selecting suitable callus for in vitro regeneration of moso bamboo. 【Method】 Three different types of callus were selected according to appearance and texture by using the immature embryo of moso bamboo as an explant to induce callus. The proliferation and regeneration ability of three types of moso bamboo callus were investigated by paraffin section and flow cytometry. Real-time PCR (qRT-PCR) was used to analyze the expression of genes related to embryonic maintenance and growth regulation in different callus types, and to select candidate genes that could be used to improve regeneration efficiency in the future. 【Result】 The type Ⅱ of white and dense callus had a large number of embryogenic meristem cells on the surface and inside. The proportion of cells in stage S and G2 in this callus type was significantly higher than that in the other types of callus, reaching 8.78% and 8.95% of the total number of cells, respectively. At the same time, the expression of two PeWUS was higher in type Ⅱ and type Ⅱ callus with strong embryogenesis, and the expression of seven PeGRF genes was the highest in type Ⅱ callus. 【Conclusion】 There were differences in proliferation and regeneration ability of different types of moso bamboo callus. The type Ⅱ of white dense callus with stronger proliferation and regeneration ability was more suitable as a tissue culture material. The selected differential genes could be used as candidate genes to improve regeneration efficiency of moso bamboo.

Key words: moso bamboo, callus, histology, cell proliferation, gene expression

CLC Number:

S718

HUANG Biyun, ZHUO Renying, QIAO Guirong. Comparative analysis of different types of callus in moso bamboo[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2023, 47(6): 141-149.

Figures/Tables 4

Table 1

Fig. 1

Fig. 2

Fig. 3

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基因 gene	基因ID gene ID	正向引物(5'-3') forward primer(5'-3')	反向引物(5'-3') reverse primer(5'-3')
	PH02Gene04456.t1	TGGAGTGGGACAATGCCAAG	ATGGAGATCTGCTTGCGGAG
	PH02Gene03707.t1	TTTTACTGCGCCAAGAACGC	TCGTCCGTCATCACCTTGAC
WUS	PH02Gene44184.t1	GCGACCATCACGGTGTTCAT	CATACTCGTTGACGGGGAGG
	PH02Gene15929.t1	GGCAGATGGGGTTTGTGGAA	CGAACATGGCTCGCAAGTCT
	PH02Gene32607.t1	AGGCACTCCACTGCACCTGT	CCAGAATGGACACCGAAGAA
	PH02Gene02258.t2	TGGGTACGGGTCCTACTTCG	TGCGTTTCCACAGGCTTTCT
	PH02Gene06385.t2	ATGACCAGGCTTACATTGCG	GTCCGTCGTCAAGTGTTTCC
	PH02Gene10190.t3	GAGCTACCCTCTGCTTGGAA	AGGAGTGGCTGTTGTCATCG
	PH02Gene23215.t2	CGTCGCAGATTCATGTCGAT	GTGATGGCTCAAGGTGGGA
	PH02Gene26399.t1	CCTCGTTTCATCGGATTCAC	CCTGGGCCAGTCCACATAGT
	PH02Gene29026.t2	AAACACTAGGGGCTTCACCA	TGACGGCTCAAGGTAGGAAT
GRF	PH02Gene34199.t1	CACGCCTTCTTCTCCGACAG	TTCGTCGTCCTCTTCTTTGG
	PH02Gene01484.t1	GCTCCCCCGCCTCTCCTGT	CGGCGAAGTAGCGGTAGATG
	PH02Gene03343.t1	ACTCGCCGCAGCCACCGT	CAAGCCCAGCCCAAGAACCAG
	PH02Gene03768.t1	GTCAACTCGCTTCCAAAGGTCG	TGGCAACGGAGAAGTCAGAGG
	PH02Gene06203.t1	CGGACGGACGGCAAGAAATG	TGCTGCTGATGGTTGATGGTGAG
	PH02Gene08777.t1	TGTGAAGCAGGAGAACCAGACGC	GAAGTCAGAGGACGCCATCGGTA
	PH02Gene33803.t1	CATCAAATGAAGGAAACAGTGCG	AAGGCGAGGGGCAACTTTTATT
	PH02Gene45969.t1	CCGCAGCATCAGCAGCACTT	GATGGGTAGGCACAGGAACACG
内参reference gene	PeTIP41	AAAATCATTGTAGGCCATTGTCG	ACTAAATTAAGCCAGCGGGAGTG

Comparative analysis of different types of callus in moso bamboo

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