Molecular Cloning and Characterization of an Acetolactate Synthase Gene（CeALS）from Cyperus esculentus L.

doi:10.13560/j.cnki.biotech.bull.1985.2021-1198

Abstract

Abstract:

Acetolactate synthase（ALS）plays an important role in the herbicide resistance of plants. To uncover the sequence characteristics,evolutionary relationship,and expression pattern of the ALS gene in tigernut（Cyperus esculentus L.）,in this study,RT-PCR was adopted to isolate the corresponding gene CeALS on the basis of transcriptome data. Sequence analysis revealed that CeALS putatively encoded 646 amino acids with a theoretical molecular weight of 69.94 kD and an isolectric point of 6.10,which was predicted to be a hydrophilic protein with chloroplast localization. The protein was shown to harbor three conserved domains,i.e.,TPP_enzyme_N,TPP_enzyme_M,and TPP_enzyme_C,which were specific to acetolactate synthase. Homologous analysis suggested that CeALS shared an identity of >90% with homologs from other Cyperaceae plants. Phylogenetic analysis supported that Cyperaceae was a sister family to Poaceae within Poales. qRT-PCR analysis revealed that CeALS was mainly expressed in mature leaves and tubers. During leaf development,an increasing trend was observed and the expression level in mature and senescent leaves was significantly higher than that in young leaves. Sequence alignment and SNP analysis supported that no target site resistance mutations to acetolactate-synthase-inhibiting herbicides were found in 56 germplasms collected by our group. Results obtained in this study lay a solid foundation for molecular breeding for herbicide resistance as well as development and utilization of tigernut.

Key words: Cyperus esculentus L., oil crop, acetolactate synthase, herbicide resistance

XIAO Yan-hua, ZOU Zhi, ZHAO Yong-guo, GUO An-ping, ZHANG Li. Molecular Cloning and Characterization of an Acetolactate Synthase Gene（CeALS）from Cyperus esculentus L.[J]. Biotechnology Bulletin, 2022, 38(4): 184-192.

Figures/Tables 7

References 26

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基因 Gene	内含子数 Number of intron	蛋白 Protein/AA	信号肽 Transit peptide/AA	分子量 MW/kD	等电点 pI	总平均疏水指数 GRAVY	脂肪族指数 AI	不稳定系数 II
AtALS	0	670	55	72.59	6.20	-0.130	87.78	42.93
PeALS	0	657	58	71.51	6.78	-0.126	95.16	40.20
PdALS	0	658	68	70.91	6.17	-0.054	95.06	43.27
EgALS	0	658	68	70.89	6.27	-0.063	94.92	41.18
SpALS	0	659	52	71.81	5.97	-0.138	92.67	43.44
AcALS	0	655	59	70.44	6.37	-0.061	93.89	45.85
MaALS1	0	655	58	71.12	6.64	-0.091	93.11	38.24
MaALS2	0	660	54	71.84	6.43	-0.103	93.03	37.67
OsALS1	0	644	43	69.39	6.48	-0.027	92.13	41.61
OsALS2	0	663	79	71.18	6.30	-0.111	87.59	36.30
HvALS	0	646	46	69.35	6.58	-0.004	92.32	40.49
BdALS1	0	646	46	69.45	6.55	-0.015	92.46	43.17
BdALS2	0	646	46	70.12	6.41	-0.066	92.15	37.31
SvALS	0	643	41	69.39	6.45	-0.053	88.96	42.81
SiALS	0	643	41	69.38	6.45	-0.054	88.96	42.84
SbALS	0	641	40	69.05	6.42	0.000	92.14	40.01
ZmALS1	0	638	39	68.93	6.69	-0.060	91.16	40.26
ZmALS2	0	638	39	68.99	6.77	-0.036	91.33	40.15
ClALS1	1	644	79	70.11	6.45	-0.145	89.35	38.09
ClALS2	1	644	79	70.34	6.80	-0.163	88.59	37.94
CbALS	1	647	55	70.46	5.85	-0.166	88.92	46.54
CeALS	1	645	54	69.94	6.10	-0.156	89.36	44.93
SwALS1	1	645	80	70.24	6.24	-0.144	90.59	41.67
SwALS2	1	645	77	70.26	6.45	-0.163	89.98	41.97
SjALS1	1	645	80	70.22	6.24	-0.140	90.74	41.67
SjALS2	1	645	80	70.26	6.45	-0.163	89.98	41.97
SmALS1	1	648	80	70.58	6.22	-0.143	90.62	41.68
SmALS2	1	645	77	70.21	6.45	-0.157	90.28	41.37

基因 Gene	内含子数 Number of intron	蛋白 Protein/AA	信号肽 Transit peptide/AA	分子量 MW/kD	等电点 pI	总平均疏水指数 GRAVY	脂肪族指数 AI	不稳定系数 II
AtALS	0	670	55	72.59	6.20	-0.130	87.78	42.93
PeALS	0	657	58	71.51	6.78	-0.126	95.16	40.20
PdALS	0	658	68	70.91	6.17	-0.054	95.06	43.27
EgALS	0	658	68	70.89	6.27	-0.063	94.92	41.18
SpALS	0	659	52	71.81	5.97	-0.138	92.67	43.44
AcALS	0	655	59	70.44	6.37	-0.061	93.89	45.85
MaALS1	0	655	58	71.12	6.64	-0.091	93.11	38.24
MaALS2	0	660	54	71.84	6.43	-0.103	93.03	37.67
OsALS1	0	644	43	69.39	6.48	-0.027	92.13	41.61
OsALS2	0	663	79	71.18	6.30	-0.111	87.59	36.30
HvALS	0	646	46	69.35	6.58	-0.004	92.32	40.49
BdALS1	0	646	46	69.45	6.55	-0.015	92.46	43.17
BdALS2	0	646	46	70.12	6.41	-0.066	92.15	37.31
SvALS	0	643	41	69.39	6.45	-0.053	88.96	42.81
SiALS	0	643	41	69.38	6.45	-0.054	88.96	42.84
SbALS	0	641	40	69.05	6.42	0.000	92.14	40.01
ZmALS1	0	638	39	68.93	6.69	-0.060	91.16	40.26
ZmALS2	0	638	39	68.99	6.77	-0.036	91.33	40.15
ClALS1	1	644	79	70.11	6.45	-0.145	89.35	38.09
ClALS2	1	644	79	70.34	6.80	-0.163	88.59	37.94
CbALS	1	647	55	70.46	5.85	-0.166	88.92	46.54
CeALS	1	645	54	69.94	6.10	-0.156	89.36	44.93
SwALS1	1	645	80	70.24	6.24	-0.144	90.59	41.67
SwALS2	1	645	77	70.26	6.45	-0.163	89.98	41.97
SjALS1	1	645	80	70.22	6.24	-0.140	90.74	41.67
SjALS2	1	645	80	70.26	6.45	-0.163	89.98	41.97
SmALS1	1	648	80	70.58	6.22	-0.143	90.62	41.68
SmALS2	1	645	77	70.21	6.45	-0.157	90.28	41.37

位点Site	CeALS	SNP	位点Site	CeALS	SNP	位点Site	CeALS	SNP
120	A	C	435	C	G	1431	T	G
147	C	T	519	C	T	1512	T	A
237	G	A	631	A	C	1596	T	C
255	G	A	681	T	C	1612	T	G
405	A	G	685	C	A	1702	A	G
408	T	C	840	C	T	1749	T	C
411	C	G	945	C	T	1752	T	C
414	C	T	1128	C	T	1767	T	C
417	C	T	1146	G	A	1771	G	T
420	C	A	1404	G	A	1851	G	T

位点Site	CeALS	SNP	位点Site	CeALS	SNP	位点Site	CeALS	SNP
120	A	C	435	C	G	1431	T	G
147	C	T	519	C	T	1512	T	A
237	G	A	631	A	C	1596	T	C
255	G	A	681	T	C	1612	T	G
405	A	G	685	C	A	1702	A	G
408	T	C	840	C	T	1749	T	C
411	C	G	945	C	T	1752	T	C
414	C	T	1128	C	T	1767	T	C
417	C	T	1146	G	A	1771	G	T
420	C	A	1404	G	A	1851	G	T