Expression Amount of cry2A* Gene at Different Growth and Development Stages of Transgenic Pest-resistant Rice ‘HD2’

doi:10.11924/j.issn.1000-6850.casb2021-0070

Abstract

Abstract:

The purpose is to investigate the expressing amount of cry2A* gene and brown rice Bt protein content (BPC) in transgenic cry2A* Gene Resistant Rice ‘HD2-1’ and other independent lines during the growth and development stages. qRT-PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the expression amount of cry2A*gene of organs at different growth and development stages and brown rice BPC. The results showed that there was a significant difference among rice independent lines in expression amount of cry2A* gene and BPC of brown rice. There were often positive correlation of the expression amount of cry2A* gene of leaf blade, sheath-culm and young panicle among different growth and development stages. The mRNA expression of cry2A* gene was positively correlated with the corresponding BPC. And there was a positive correlation trend between BPC of brown rice and expression amount of cry2A* gene of each organ at each stage. In expression amount of cry2A* gene of rice‘HD2’, there were positive relations among different growth and development stages and between the ones and BPC of brown rice. The higher the cry2A* gene transcribed, the higher the BPC was.

Key words: transgenic pest-resistant rice, ‘Kongyu 131’, growth and development stage, expression amount of cry2A* gene, Bt protein content (BPC)

CLC Number:

S511.22

Li Rongtian, Yu Xiaochen, Kong Mengying, Liu Changhua. Expression Amount of cry2A* Gene at Different Growth and Development Stages of Transgenic Pest-resistant Rice ‘HD2’[J]. Chinese Agricultural Science Bulletin, 2021, 37(23): 15-22.

Figures/Tables 8

References 26

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引物名称	正向引物	反向引物	产物大小/bp
actin	5′-TATGGTCAAGGCTGGGTTCG-3′	5′-CCATGCTCGATGGGGTACTT-3′	177
cry2A*	5′-CGTGTCAATGCTGACCTGAT-3′	5′-GATGCCGGACAGGATGTAGT-3′	600
bar	5′-TCTCGGTGACGGGCAGGAC-3′	5′-CCTGGAAGGCACGCAACG-3′	323

引物名称	正向引物	反向引物	产物大小/bp
actin	5′-TATGGTCAAGGCTGGGTTCG-3′	5′-CCATGCTCGATGGGGTACTT-3′	177
cry2A*	5′-CGTGTCAATGCTGACCTGAT-3′	5′-GATGCCGGACAGGATGTAGT-3′	600
bar	5′-TCTCGGTGACGGGCAGGAC-3′	5′-CCTGGAAGGCACGCAACG-3′	323

阶段	器官	mRNA表达量				F值	差异显著性
阶段	器官	HD2-1	HD2-2	HD2-3	HD2-4	F值	差异显著性
分蘖期	叶片	351.433±1.442	347.650±1.198	254.240±1.691	223.370±1.294	4222.46	**
分蘖期	茎鞘	16.770±1.192	16.227±1.394	10.343±1.115	8.780±1.120	22.52	**
抽穗期	叶片	787.173±1.105	778.110±1.478	679.437±1.477	547.770±1.155	14370.18	**
	茎鞘	31.517±1.499	31.413±1.484	27.360±0.441	26.960±0.148	43.42	**
	幼穗	124.370±1.070	118.210±0.952	67.457±1.004	61.423±1.021	2125.70	**
灌浆期	叶片	986.697±1.701	935.810±1.232	753.787±1.217	715.120±1.457	17783.56	**
	茎鞘	38.890±0.270	38.087±1.183	36.063±1.379	34.900±1.125	5.77	*
	幼穗	28.507±0.977	23.260±1.006	16.317±1.050	13.357±1.047	100.00	**

阶段	器官	mRNA表达量				F值	差异显著性
阶段	器官	HD2-1	HD2-2	HD2-3	HD2-4	F值	差异显著性
分蘖期	叶片	351.433±1.442	347.650±1.198	254.240±1.691	223.370±1.294	4222.46	**
分蘖期	茎鞘	16.770±1.192	16.227±1.394	10.343±1.115	8.780±1.120	22.52	**
抽穗期	叶片	787.173±1.105	778.110±1.478	679.437±1.477	547.770±1.155	14370.18	**
	茎鞘	31.517±1.499	31.413±1.484	27.360±0.441	26.960±0.148	43.42	**
	幼穗	124.370±1.070	118.210±0.952	67.457±1.004	61.423±1.021	2125.70	**
灌浆期	叶片	986.697±1.701	935.810±1.232	753.787±1.217	715.120±1.457	17783.56	**
	茎鞘	38.890±0.270	38.087±1.183	36.063±1.379	34.900±1.125	5.77	*
	幼穗	28.507±0.977	23.260±1.006	16.317±1.050	13.357±1.047	100.00	**

阶段	器官	BPC/(μg/g)				F值	差异显著性
阶段	器官	HD2-1	HD2-2	HD2-3	HD2-4	F值	差异显著性
分蘖期	叶片	14.263±0.113	14.197±0.135	13.663±0.180	12.910±0.106	68.53	**
分蘖期	茎鞘	1.870±0.104	1.740±0.089	1.160±0.090	1.023±0.084	41.28	**
抽穗期	叶片	38.657±0.180	36.207±0.125	34.940±0.340	29.877±0.069	638.97	**
	茎鞘	4.833±0.073	4.287±0.108	3.860±0.067	3.437±0.132	73.43	**
	幼穗	4.363±0.054	4.107±0.140	3.693±0.147	3.420±0.164	19.98	**
灌浆期	叶片	48.563±0.151	44.693±0.125	38.240±0.057	35.657±0.164	4071.18	**
	茎鞘	5.840±0.051	4.920±0.119	4.327±0.070	3.977±0.100	168.33	**
	幼穗	2.860±0.156	2.213±0.092	1.857±0.066	1.633±0.074	53.97	**
完熟期	糙米	0.727±0.074	0.577±0.037	0.413±0.012	0.293±0.074	22.94	**