Analysis of SHMT Gene Family and Production of PtrSHMT9 Mutant in Populus trichocarpa

doi:10.7525/j.issn.1673-5102.2020.06.013

Abstract

Abstract:

Serine hydroxy methyltransferases（SHMTs） played an important role in the one-carbon（C1） metabolism pathway in plant cells. We identified 9 SHMT genes from the genome of Populus trichocarpa. Populus eFP resource showed that 7 PtrSHMTs were expressed in multiple tissues， among which PtrSHMT9 was the most highly expressed gene in xylem. By analysis of RT-PCR and Aspwood， PtrSHMT9 had a high level in secondary wall thickening of stem wood tissue， suggested its involvement in poplar wood formation. Using Cas9/gRNA technique， PtrSHMT9-edited mutants were prepared and 4 different ptrshmt9 knockouts were obtained. Our results provided basic information and genetic material for exploring the function of SHMT and PtrSHMT9 in trees.

Key words: Populus trichocarpa, SHMT, ptrshmt9mutant, Cas9/gRNA

CLC Number:

S792.11

Bing LI, Yu-Xiang CHENG. Analysis of SHMT Gene Family and Production of PtrSHMT9 Mutant in Populus trichocarpa[J]. Bulletin of Botanical Research, 2020, 40(6): 906-912.

Figures/Tables 8

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引物名称 Primer name	引物用途 Primer use	引物序列 Primer sequence(5′—3′)
PtrSHMT9-RT	半定量RT-PCR Semi-quantitative PCR	CCTTGTTGCTGCTCAGGAAGCTG; CAGAGCACCAATCTGGTGGTTGTG
DT1-BsF	靶位点gRNA Target site gRNA	ATATATGGTCTCGATTGGGCTTGGATTTGCCTTCAGGTT
DT1-F0		TGGGCTTGGATTTGCCTTCAGGTTTTAGAGCTAGAAATAGC
DT2-R0		AACGCCCTATTCTGGTTCCCCTGAATCTCTTAGTCGACTCTAC
DT2-BsR		ATTATTGGTCTCTAAACGCCCTATTCTGGTTCCCCTG
PtrSHMT9-U/D	位点编辑鉴定 Identifying editing site	ATGGATCCAGTAACTGTGTGG; CCACTGATGTGAGCCATGTC
zCas-U/D	转基因鉴定 Transgenic identification	TGAGAACATCGTCATTGAGATGG; TCAGCTTGTCATTCTCATCGTAC
PtrActin2-U/D	Actin内参基因 Reference gene Actin	AACATGGGATTGTTAGCAACTGG; TCCATCACCAGAATCCAGCACA

基因名 Gene name	基因座位 Gene locus	氨基酸数目 Amino acid
PtrSHMT1	Potri.001G212000	578
PtrSHMT2	Potri.001G320400	471
PtrSHMT3	Potri.002G090200	555
PtrSHMT4	Potri.002G109200	516
PtrSHMT5	Potri.005G170800	552
PtrSHMT6	Potri.006G232300	559
PtrSHMT7	Potri.008G002900	547
PtrSHMT8	Potri.010G254700	520
PtrSHMT9	Potri.017G059300	471

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