春兰miR396过表达对拟南芥叶片生长、光合及叶绿素荧光特性的影响

doi:10.13560/j.cnki.biotech.bull.1985.2020-1203

摘要/Abstract

摘要：

为研究春兰miR396（cgo-miR396）基因在植物叶片生长发育中的作用,针对春兰的栽培和生产提供新的研究思路。将该基因前体cgo-MIR396在拟南芥中过量表达,观察其形态指标的变化并对相应的光合和叶绿素荧光参数进行测定。春兰miR396基因在花期叶片中的表达量最高,过表达该基因可以显著增加拟南芥的叶长、叶宽、叶面积和株高,并明显降低叶片的叶绿素含量、气孔导度（gs）和蒸腾速率（Tr）,提高了水分利用效率（WUE）;另外,cgo-miR396的过表达还导致反应中心吸收的光能用于电子传递的量子产额（φEo）、性能指数（PI_ABS）、推动力（DF_ABS）等叶绿素荧光参数的下降。春兰miR396主要于生殖生长时期发挥作用,在保证净光合效率和叶片生长发育的同时,适当降低了叶片的气孔导度及PSII受体侧的部分性能。

关键词: 春兰, miR396, 拟南芥, 光合, 叶绿素荧光

Abstract:

The objective of this work is to study the role of Cymbidium goeringii’s miR396（cgo-miR396）in the growth and development of plant leaves,and to provide a new research idea for the cultivation and production of C. goeringii. The precursor of gene cgo-MIR396 was overexpressed in Arabidopsis thaliana,and the changes of its morphological indexes were observed and the corresponding photosynthetic and chlorophyll fluorescence parameters were determined. As results,gene cgo-miR396 was in the highest expression level in leaves at flowering stage,and the overexpression of cgo-miR396 significantly increased the leaf length,leaf width,leaf area and plant height of Arabidopsis thaliana,while significantly reduced the chlorophyll content,stomatal conductance（gs）and transpiration rate（Tr）of leaves,and improved the water use efficiency（WUE）. In addition,the overexpression of this gene also resulted in the decrease in the quantum yield（φEo）,performance index（PI_ABS）,thrust force（DF_ABS）and other chlorophyll fluorescence parameters of the light absorbed by the reaction center for electron transfer. In conclusion,cgo-miR396 mainly plays a role in the reproductive growth period and appropriately reduces stomatal conductance of plant leaves and some properties of the PSII receptor side while the net photosynthetic efficiency of plants and promoting leaf growth is ensured.

Key words: Cymbidium goeringii, miR396, Arabidopsis thaliana, photosynthesis, chlorophyll fluorescence

徐子涵, 刘倩, 苗大鹏, 陈跃, 胡凤荣. 春兰miR396过表达对拟南芥叶片生长、光合及叶绿素荧光特性的影响[J]. 生物技术通报, 2021, 37(5): 28-37.

XU Zi-han, LIU Qian, MIAO Da-peng, CHEN Yue, HU Feng-rong. Impacts of Cymbidium goeringii’s miR396 Overexpression on the Leaf Growth,Photosynthesis and Chlorophyll Fluorescence in Arabidopsis thaliana[J]. Biotechnology Bulletin, 2021, 37(5): 28-37.

图/表 11

参考文献 16

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株系	叶绿素a	叶绿素b	叶绿素 a+b	叶绿素a/b
Line	Chla/（mg·g^-1）	Chlb/（mg·g^-1）	Chl（a+b）/（mg·g^-1）	Chla/b
WT	1.797±0.09a	0.467±0.02a	2.267±0.11a	3.853±0.04a
Line1	1.300±0.09b	0.340±0.03b	1.637±0.12b	3.883±0.13a
Line2	1.280±0.10b	0.330±0.04b	1.607±0.14b	3.923±0.18a
Line3	1.300±0.06b	0.337±0.02b	1.640±0.08b	3.857±0.06a
Line4	1.497±0.05b	0.397±0.03ab	1.893±0.08b	3.770±0.12a

株系	叶绿素a	叶绿素b	叶绿素 a+b	叶绿素a/b
Line	Chla/（mg·g^-1）	Chlb/（mg·g^-1）	Chl（a+b）/（mg·g^-1）	Chla/b
WT	1.797±0.09a	0.467±0.02a	2.267±0.11a	3.853±0.04a
Line1	1.300±0.09b	0.340±0.03b	1.637±0.12b	3.883±0.13a
Line2	1.280±0.10b	0.330±0.04b	1.607±0.14b	3.923±0.18a
Line3	1.300±0.06b	0.337±0.02b	1.640±0.08b	3.857±0.06a
Line4	1.497±0.05b	0.397±0.03ab	1.893±0.08b	3.770±0.12a

株系	初始荧光	最大荧光	可变荧光	PS II最大光化学效率	PS II 潜在光化学效率
Line	F_o	F_m	F_v	F_v/F_m	F_v/F_o
WT	547.5±9.93a	3229.5±35.62a	2682.0±27.04a	0.831±0.002a	4.905±0.05a
Line1	541.7±26.21a	3252.3±115.61a	2710.7±104.80a	0.833±0.007a	5.023±0.27a
Line2	537.5±0.29a	3115.5±79.39a	2578.0±79.10a	0.828±0.004a	4.796±0.14a
Line3	564.7±21.36a	3104.3±17.64a	2539.7±5.46a	0.818±0.006a	4.511±0.18a
Line4	551.0±8.89a	3282.0±96.16a	2731.0±87.47a	0.832±0.002a	4.954±0.08a

株系	初始荧光	最大荧光	可变荧光	PS II最大光化学效率	PS II 潜在光化学效率
Line	F_o	F_m	F_v	F_v/F_m	F_v/F_o
WT	547.5±9.93a	3229.5±35.62a	2682.0±27.04a	0.831±0.002a	4.905±0.05a
Line1	541.7±26.21a	3252.3±115.61a	2710.7±104.80a	0.833±0.007a	5.023±0.27a
Line2	537.5±0.29a	3115.5±79.39a	2578.0±79.10a	0.828±0.004a	4.796±0.14a
Line3	564.7±21.36a	3104.3±17.64a	2539.7±5.46a	0.818±0.006a	4.511±0.18a
Line4	551.0±8.89a	3282.0±96.16a	2731.0±87.47a	0.832±0.002a	4.954±0.08a

	叶面积	叶长	叶宽	株高	叶绿素a+b	叶绿素a/b	Ci	gs	A	E	WUE	F_o	F_m	F_v	F_v/F_m	F_v/F_o	ABS/RC	DIo/RC	TRo/RC	ETo/RC	φPo	ψo	φEo	PI_ABS	DF _ABS
叶面积	1
叶长	0.883^**	1
叶宽	0.953^**	0.733^**	1
株高	0.464	0.470	0.408	1
叶绿素a+b	-0.194	-0.046	-0.284	-0.281	1
叶绿素a/b	0.017	-0.239	0.179	-0.037	-0.257	1
Ci	-0.206	-0.361	-0.154	-0.299	0.290	0.471	1
gs	-0.551^*	-0.579^*	-0.524^*	-0.269	0.426	0.363	0.778^**	1
A	-0.004	-0.033	0.070	0.139	-0.049	-0.153	-0.072	-0.018	1
E	-0.412	-0.488	-0.364	-0.332	0.326	0.312	0.868^**	0.890^**	0.087	1
WUE	0.261	0.384	0.227	0.419	-0.279	-0.420	-0.922^**	-0.797^**	0.177	-0.921^**	1
F_o	0.143	0.268	0.118	0.034	0.004	0.178	0.218	0.248	0.360	0.214	-0.169	1
F_m	-0.137	-0.029	-0.254	-0.199	0.291	-0.032	0.261	0.233	0.286	0.226	-0.173	0.327	1
F_v	-0.171	-0.083	-0.289	-0.215	0.304	-0.068	0.230	0.196	0.229	0.195	-0.148	0.146	0.983^**	1
F_v/F_m	-0.249	-0.276	-0.323	-0.191	0.227	-0.177	0.030	-0.015	-0.085	0.007	-0.004	-0.629^*	0.529^*	0.677^**	1
F_v/F_o	-0.256	-0.289	-0.319	-0.195	0.237	-0.183	0.029	-0.027	-0.062	-0.003	0.011	-0.636^*	0.518^*	0.667^**	0.996^**	1
ABS/RC	0.346	0.408	0.352	0.271	-0.243	0.052	-0.308	-0.314	0.007	-0.453	0.407	0.419	-0.177	-0.267	-0.516^*	-0.510	1
DIo/RC	0.350	0.369	0.417	0.239	-0.267	0.240	-0.110	-0.115	0.057	-0.193	0.153	0.631^*	-0.409	-0.552^*	-0.901^**	-0.893^**	0.824^**	1
TRo/RC	0.302	0.377	0.278	0.253	-0.202	-0.044	-0.366	-0.371	-0.018	-0.522^*	0.478	0.266	-0.044	-0.098	-0.269	-0.264	0.962^**	0.639^*	1
ETo/RC	-0.049	0.092	-0.164	-0.348	0.216	-0.159	-0.140	-0.033	-0.227	-0.287	0.191	0.218	0.369	0.343	0.111	0.090	0.461	0.138	0.560^*	1
φPo	-0.273	-0.261	-0.368	-0.167	0.222	-0.325	-0.055	-0.058	-0.075	-0.040	0.066	-0.648^**	0.490	0.640^*	0.986^**	0.981^**	-0.506	-0.905^**	-0.252	0.129	1
ψo	-0.277	-0.159	-0.398	-0.607^*	0.405	-0.159	0.098	0.228	-0.259	0.032	-0.116	0.080	0.476	0.482	0.319	0.291	-0.123	-0.281	-0.033	0.810^**	0.330	1
φEo	-0.309	-0.196	-0.439	-0.592^*	0.415	-0.211	0.078	0.197	-0.250	0.021	-0.092	-0.050	0.533^*	0.568^*	0.484	0.457	-0.212	-0.434	-0.079	0.769^**	0.496	0.983^**	1
PI_ABS	-0.343	-0.357	-0.402	-0.624^*	0.430	0.075	0.310	0.301	-0.214	0.233	-0.314	-0.240	0.605^*	0.680^**	0.710^**	0.697^**	-0.559^*	-0.707^**	-0.420	0.395	0.657^**	0.773^**	0.838^**	1
DF _ABS	-0.313	-0.334	-0.379	-0.580^*	0.430	0.070	0.302	0.285	-0.237	0.211	-0.297	-0.267	0.596^*	0.676^**	0.727^**	0.712^**	-0.561^*	-0.721^**	-0.416	0.394	0.676^**	0.768^**	0.837^**	0.997^**	1