Functional analysis of glycine decarboxylase H-protein gene MsGDC-H1 in Medicago sativa

doi:10.11686/cyxb2021477

Abstract

Abstract:

Photorespiration is essential for C₃ plants， allowing oxygenated photosynthesis by removing 2-phosphoglycolate. H-protein is one of the key components of glycine decarboxylase （GDC）， which converts glycine to serine during photorespiration. The MsGDC-H1 gene was cloned from Medicago sativa. The cloned gene encoded 166 amino acids and had a conserved lipoyl-binding domain and a conserved N6-lipoyllysine site. Phylogenetic analysis showed that MsGDC-H1 was closely related to glycine decarboxylase H-protein （GDC-H） in other dicotyledons. The expression pattern analysis showed that MsGDC-H1 was highly expressed in leaves and was induced by light. To explore the effects of MsGDC-H1 on Arabidopsis thaliana growth， photoinduced stem-leaf specific promoter ST-LS1 and constitutive promoter CaMV 35S were used to drive MsGDC-H1 expression （ST-LS1：：MsGDC-H1； CaMV 35S：： MsGDC-H1）. MsGDC-H1 is heterogeneously expressed in A. thaliana. Biomass， starch and soluble sugar content and photosynthetic rate of overexpressed plants were evaluated. Data analysis showed that CaMV 35S：：MsGDC-H1 overexpression （G series） inhibited A. thaliana growth. The starch content of G series was increased by 34%-67% compared with A. thaliana in which ST-LS1：：MsGDC-H1 was overexpressed （GS series）， and was increased by 7.3%-33.7% compared with wild type （WT） A. thaliana； The soluble sugar content was decreased by 36%-38% compared with GS series and increased by 44.3%-49.7% compared with WT A. thaliana. Compared with WT， GS series plants grew faster， starch content was not significantly different （P>0.05）， and soluble sugar content was significantly increased （P<0.05）. The experiment results show that MsGDC-H1 plays an important role in regulating photosynthetic rate， carbohydrate synthesis and growth of A. thaliana， and can be used as a candidate gene in genetic engineering breeding to improve alfalfa yield in the future.

Key words: glycine decarboxylase H-protein, yield, photosynthetic rate, carbohydrate content, Arabidopsis thaliana

Chun-xue WEI, Fei HE, Lei XU, Xiao LI, Li-xia ZHANG, Ming-na LI, Lin CHEN, Jun-mei KANG, Qing-chuan YANG, Rui-cai LONG. Functional analysis of glycine decarboxylase H-protein gene MsGDC-H1 in Medicago sativa[J]. Acta Prataculturae Sinica, 2022, 31(12): 95-105.

Figures/Tables 8

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引物Primer	引物序列Primer sequence （5′-3′）
MsH-F	ATGGCACTGAGGATGTGGGCTTC
MsH-R	TCAATGTGCAGCATCTTCTTCCTCA
ST-LS1-F	GCAAGCTTTTGTTTCTATCCACTGATGTCTG
ST-LS1-R	GCGGATCCTTGCTCTCACTACTTAGTATGA
qRT-MsH-F	CACTGAGGATGTGGGCTTCTT
qRT-MsH-R	AGCCTTCATGCTTCACCCAT
qRT-Atactin-F	CAAAAGATGGCAGATGCTGAGGAT
qRT-Atactin-R	CATGCACCAGTATGACGAGGTCG
qRT-Msactin-F	GAAATCACAGCACTTGCACC
qRT-Msactin-R	AAGCCTTTGATCTTGAGAGC

引物Primer	引物序列Primer sequence （5′-3′）
MsH-F	ATGGCACTGAGGATGTGGGCTTC
MsH-R	TCAATGTGCAGCATCTTCTTCCTCA
ST-LS1-F	GCAAGCTTTTGTTTCTATCCACTGATGTCTG
ST-LS1-R	GCGGATCCTTGCTCTCACTACTTAGTATGA
qRT-MsH-F	CACTGAGGATGTGGGCTTCTT
qRT-MsH-R	AGCCTTCATGCTTCACCCAT
qRT-Atactin-F	CAAAAGATGGCAGATGCTGAGGAT
qRT-Atactin-R	CATGCACCAGTATGACGAGGTCG
qRT-Msactin-F	GAAATCACAGCACTTGCACC
qRT-Msactin-R	AAGCCTTTGATCTTGAGAGC

分类 Category	物种 Species	光合类型 Photosynthetic types	数目Number
分类 Category	物种 Species	光合类型 Photosynthetic types	GDC-H	GDC-H外显子Exons
双子叶植物 Dicot	紫花苜蓿M.sativa	C₃	3	1，3，4
	蒺藜苜蓿M. truncatula	C₃	2	3，4
	拟南芥A. thaliana	C₃	3	3，4
	大豆G. max	C₃	5	3，4
	烟草N. tabacum	C₃	4	3，4
	狭叶羽扇豆L. angustifolius	C₃	4	2，3，4
单子叶植物 Monocot	水稻O. sativa	C₃	4	4，9
	玉米Z. mays	C₄	6	3，4，5，7
	凤梨A. comosus	CAM	2	3，4
	高粱S. bicolor	C₄	2	4
低等植物 Lower plants	小立碗藓P. patens	C₃	2	1，4
	江南卷柏S. moellendorffii	C₃	2	4
	莱茵衣藻C. reinhardtii	C₃	1	5

分类 Category	物种 Species	光合类型 Photosynthetic types	数目Number
分类 Category	物种 Species	光合类型 Photosynthetic types	GDC-H	GDC-H外显子Exons
双子叶植物 Dicot	紫花苜蓿M.sativa	C₃	3	1，3，4
	蒺藜苜蓿M. truncatula	C₃	2	3，4
	拟南芥A. thaliana	C₃	3	3，4
	大豆G. max	C₃	5	3，4
	烟草N. tabacum	C₃	4	3，4
	狭叶羽扇豆L. angustifolius	C₃	4	2，3，4
单子叶植物 Monocot	水稻O. sativa	C₃	4	4，9
	玉米Z. mays	C₄	6	3，4，5，7
	凤梨A. comosus	CAM	2	3，4
	高粱S. bicolor	C₄	2	4
低等植物 Lower plants	小立碗藓P. patens	C₃	2	1，4
	江南卷柏S. moellendorffii	C₃	2	4
	莱茵衣藻C. reinhardtii	C₃	1	5

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