Expressions and of Genes Response to Signal Substances in MAPK Cascade Pathway Genes in Pinus massoniana

doi:10.13560/j.cnki.biotech.bull.1985.2021-1089

Abstract

Abstract:

MAPK（mitogen-actived protein kinase）cascade pathway plays an important role in response and regulation of biotic,abiotic stresses,and hormone signals et al. In this study,7 genes related to MAPK cascade pathway in Pinus massoniana were identified. We analyzed their physical properties,functional domains,phylogenetic tree,conserved phosphorylation motifs,and gene interactions. We also tested the expression patterns of these genes under hormone and calcium treatment. The results showed that：these genes had close evolutionary relationships with gymnosperms,such as Pinus taeda and Picea asperata. These 7 genes had MAPK domain and typical catalytic amino acid motif. D（I/L/V）K was the common active motif of MAPKs genes of Pinus massoniana. PmMAP4K did not interact directly with other three members of MAPK cascade pathway. Genes in MAPK cascade might regulate the biosynthesis of various hormones by phosphorylation in response to abiotic stress. The results of gene expression showed that all the seven genes responded to MeJA,GA,SA,ABA and Ca²⁺ treatments,but the expression patterns were different. The results showed that the response of PmMAP4K to Ca²⁺ treatment was the most obvious followed by PmMAP2K gene,and other genes had no obvious response to Ca²⁺ treatment only. Four hormones and hormone-plus-Ca²⁺ treatments increased the expression of genes,respectively. The expression of PmMAPK in MeJA+Ca²⁺ treatment was higher than that in MeJA treatment alone,while the expression of genes under treatments of each GA,SA,and ABA was higher than that under treatments of each GA,SA,ABA plus Ca²⁺. This study provides theoretical support for the analysis of the interaction between MAPK cascade pathway genes and signal pathway in respond to abiotic stress.

Key words: conifer, MAPK cascade, hormone metabolism, gene regulation

CHEN Hu, YANG Zhang-qi, SUN Shuang, LI Peng, XU Hui-lan. Expressions and of Genes Response to Signal Substances in MAPK Cascade Pathway Genes in Pinus massoniana[J]. Biotechnology Bulletin, 2022, 38(6): 187-197.

Figures/Tables 10

References 33

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Gene	Forward primer（5'-3'）	Reverse primer（5'-3'）
PmCYP	CAAGGGTTCGTCGTTCCAC	TACGGCGAGAAGTTTGCC
PmMAPK2	CCATGGCGGTCAGTATGTGCAG	CATATGGCATTGTCTGCTCGGCA
PmMAPK6	CCGCAGCAGCTCCAGTTCAATC	CTCAGTCACGGCGGCCTCTT
PmMAPK9	GATCCGCAATGAGAAGGCCAGAAG	CCTTCGTCTGCTTGAGCGTATGC
PmMAPK15	CCTCATCAGAGCAGGCATCTTCAG	GAGAAGCAGCACCAAGACAGGAG
PmMAP2K	GTTGGAGTACATGGACGGTGGAAC	GGTTGCCAGACAGGTGCTGAAG
PmMAP3K	AGCAGCAGTCCTCCAGCGATC	GAGGTCTTGATTGCCGCCAACAA
PmMAP4K	ATGCTTCTCTGCCTCCTCTTCTCC	GACGATTCCGAAGTTCCGCAACA

Gene	Forward primer（5'-3'）	Reverse primer（5'-3'）
PmCYP	CAAGGGTTCGTCGTTCCAC	TACGGCGAGAAGTTTGCC
PmMAPK2	CCATGGCGGTCAGTATGTGCAG	CATATGGCATTGTCTGCTCGGCA
PmMAPK6	CCGCAGCAGCTCCAGTTCAATC	CTCAGTCACGGCGGCCTCTT
PmMAPK9	GATCCGCAATGAGAAGGCCAGAAG	CCTTCGTCTGCTTGAGCGTATGC
PmMAPK15	CCTCATCAGAGCAGGCATCTTCAG	GAGAAGCAGCACCAAGACAGGAG
PmMAP2K	GTTGGAGTACATGGACGGTGGAAC	GGTTGCCAGACAGGTGCTGAAG
PmMAP3K	AGCAGCAGTCCTCCAGCGATC	GAGGTCTTGATTGCCGCCAACAA
PmMAP4K	ATGCTTCTCTGCCTCCTCTTCTCC	GACGATTCCGAAGTTCCGCAACA

基因 Gene	开放阅读框 Open reading frame	氨基酸序列Amino acid sequence/aa	蛋白分子量Protein mass/kD	等电点 Protein isoelectric point	亚细胞定位Subcellular localization	跨膜结构Transmembrane	磷酸化位点Kinase phosphorylation（Ser,Thr,Tyr）	二级结构Secondary structure/%
基因 Gene	开放阅读框 Open reading frame	氨基酸序列Amino acid sequence/aa	蛋白分子量Protein mass/kD	等电点 Protein isoelectric point	亚细胞定位Subcellular localization	跨膜结构Transmembrane	磷酸化位点Kinase phosphorylation（Ser,Thr,Tyr）	α螺旋 Alpha helix	延伸链 Extended strand	β转角 Beta turn	无规卷曲 Random coil
PmMAPK6	1 173	390	44.49	5.38	细胞核	无	8,10,8	42.31	13.59	4.36	39.74
PmMAPK9	1 839	612	69.41	9.11	细胞核	无	44,23,8	44.77	10.46	4.25	40.52
PmMAPK2	1 119	372	42.95	5.28	细胞核	无	11,11,9	43.82	14.25	5.91	36.02
PmMAPK15	1 731	576	65.7	9.22	细胞核	无	30,16,11	38.54	12.85	5.73	42.88
PmMAP2K	1 014	337	37.62	9.02	细胞核	无	15,4,3	34.42	16.02	5.64	43.92
PmMAP3K	1 962	653	72.33	5.86	细胞核	无	56,26,7	28.18	9.8	4.44	57.58
PmMAP4K	2 748	915	100.62	5.12	细胞核	无	77,41,19	29.4	12.13	4.92	53.55

基因 Gene	开放阅读框 Open reading frame	氨基酸序列Amino acid sequence/aa	蛋白分子量Protein mass/kD	等电点 Protein isoelectric point	亚细胞定位Subcellular localization	跨膜结构Transmembrane	磷酸化位点Kinase phosphorylation（Ser,Thr,Tyr）	二级结构Secondary structure/%
基因 Gene	开放阅读框 Open reading frame	氨基酸序列Amino acid sequence/aa	蛋白分子量Protein mass/kD	等电点 Protein isoelectric point	亚细胞定位Subcellular localization	跨膜结构Transmembrane	磷酸化位点Kinase phosphorylation（Ser,Thr,Tyr）	α螺旋 Alpha helix	延伸链 Extended strand	β转角 Beta turn	无规卷曲 Random coil
PmMAPK6	1 173	390	44.49	5.38	细胞核	无	8,10,8	42.31	13.59	4.36	39.74
PmMAPK9	1 839	612	69.41	9.11	细胞核	无	44,23,8	44.77	10.46	4.25	40.52
PmMAPK2	1 119	372	42.95	5.28	细胞核	无	11,11,9	43.82	14.25	5.91	36.02
PmMAPK15	1 731	576	65.7	9.22	细胞核	无	30,16,11	38.54	12.85	5.73	42.88
PmMAP2K	1 014	337	37.62	9.02	细胞核	无	15,4,3	34.42	16.02	5.64	43.92
PmMAP3K	1 962	653	72.33	5.86	细胞核	无	56,26,7	28.18	9.8	4.44	57.58
PmMAP4K	2 748	915	100.62	5.12	细胞核	无	77,41,19	29.4	12.13	4.92	53.55

基因名/结构域Gene/Domain	MAPK	MAP2K	MAP3K	MAP4K	SPS1	PknB	BREX-PgiW
PmMAPK6	50-386				63-377	111-260
PmMAPK9	91-428				92-384	89-380
PmMAPK2	32-368				42-344	95-242
PmMAPK15	24-361				25-317	24-233	71-246
PmMAP2K		56-323			59-336	112-258	64-258
PmMAP3K			40-303		41-396	40-298
PmMAP4K				299-554	299-558	300-549