甘草4CL基因家族鉴定及其在干旱胁迫下的表达分析

doi:10.7668/hbnxb.20194561

华北农学报 ›› 2024, Vol. 39 ›› Issue (S1): 1-10. doi: 10.7668/hbnxb.20194561

所属专题： 抗旱节水；生物技术；

• 作物遗传育种·种质资源·生物技术 • 下一篇

甘草4CL基因家族鉴定及其在干旱胁迫下的表达分析

赵斌¹, 姚华¹^,², 石娜娜¹, 高转转¹, 杨茂¹, 冯江华³, 沈海涛¹

¹ 石河子大学生命科学学院,新疆植物药资源利用教育部重点实验室,绿洲城镇与山盆系统生态兵团重点实验室,新疆石河子 832003
² 新疆维吾尔自治区药物研究所,新疆乌鲁木齐 831004
³ 新疆师范大学商学院,新疆乌鲁木齐 830054

收稿日期:2024-05-26 出版日期:2025-01-24
通讯作者:
冯江华(1978-),男,新疆库尔勒人,副教授,博士,主要从事区域经济学研究。沈海涛(1980-),男,新疆库尔勒人,教授,博士,主要从事新疆特色药用植物资源开发与综合利用研究。
冯江华(1978-),男,新疆库尔勒人,副教授,博士,主要从事区域经济学研究。沈海涛(1980-),男,新疆库尔勒人,教授,博士,主要从事新疆特色药用植物资源开发与综合利用研究。
作者简介:
赵斌(1998-),男,新疆昌吉人,主要从事甘草抗逆品种的育种研究。
赵斌、姚华为同等贡献作者。
基金资助:
国家自然科学基金项目(32260083); 兵团财政科技计划项目(2020AA005); 兵团重点领域科技攻关项目-基础研究项目(2023CB008-17); 高层次人才科研启动经费项目(RCZK2021B31); 石河子大学新品种培育专项(YZZX202102)

Identification of 4CL Gene Family in Glycyrrhiza uralensis and Its Expression under Drought Stress

ZHAO Bin¹, YAO Hua¹^,², SHI Nana¹, GAO Zhuanzhuan¹, YANG Mao¹, FENG Jianghua³, SHEN Haitao¹

¹ College of Life Sciences,Shihezi University,Key Laboratory of Xinjiang Phytomedicine Resource and Utilization,Ministry of Education,Key Laboratory of Ecological Corps for Oasis City and Mountain Basin System,Shihezi 832003,China
² Xinjiang Uygur Autonomous Region Institute of Materia Medica,Urumqi 831004,China
³ Business School,Xinjiang Normal University,Urumqi 830054,China

Received:2024-05-26 Published:2025-01-24

摘要/Abstract

摘要：

4-香豆酸:辅酶A连接酶(4CL)作为类黄酮与木质素的主要生物合成酶,与植物黄酮类化合物的形成存在紧密的联系。为了探索甘草4CL基因家族与异甘草素的合成、积累的关系,测定甘草在干旱胁迫后,异甘草素的含量,及甘草4CL基因家族表达特性和生物信息学分析,了解异甘草素积累特性与甘草4CL基因的关系。结果显示,异甘草素主要积累在甘草根中,且干旱胁迫能显著提升甘草根中的异甘草素含量,在聚乙二醇(PEG)胁迫2 h后的含量是对照组(0 h)的3.91倍。干旱胁迫可诱导地下部分中的Gu4CL基因上调,Gu4CL2、Gu4CL4和Gu4CL5经PEG胁迫诱导后表达量较高,而Gu4CL2在PEG胁迫后上调最为明显,Gu4CL2表达与异甘草素含量变化相似。Gu4CL基因的生物信息学分析显示,11个基因都有2个保守的多肽基序BoxⅠ(SSGTTGLPKGV)和BoxⅡ(GEICIRG),并且11个基因分布在11个Scaffold片段上。启动子顺式作用元件分析发现,Gu4CL2、Gu4CL4和Gu4CL5基因较其他Gu4CL基因包含更多的脱落酸和茉莉酸响应元件。因此,干旱胁迫可能诱导茉莉酸、脱落酸的合成,调控甘草根中Gu4CL2、Gu4CL4和Gu4CL5基因的表达,提升异甘草素的积累。

关键词: 甘草, Gu4CL, 干旱胁迫, 异甘草素, 生物信息学分析

Abstract:

4-coumaric acid:coenzyme A ligase(4CL),as the main biosynthetic enzyme of flavonoids and lignin,is closely related to the formation of plant flavonoids.In order to explore the relationship between the 4CL gene family of Glycyrrhiza uralensis and the synthesis and accumulation of isoglycyrrhizin,this study determined the content of isoglycyrrhizin in G. uralensis after drought stress,and the expression characteristics and bioinformatics analysis of the 4CL gene family of G. uralensis Fisch.to understand the relationship between the accumulation characteristics of isoglycyrrhizin and the 4CL gene of G.uralensis.The results showed that isoliquiritigenin was mainly accumulated in the roots of G.uralensis,and drought stress could significantly increase the content of isoliquiritigenin in the roots of G.uralensis.The content of isoliquiritigenin after 2 h of polyethylene glycol(PEG)stress was 3.91 times that of the control group(0 h).Drought stress could induce the up regulation of Gu4CL gene in the underground part.The expression levels of Gu4CL2, Gu4CL4 and Gu4CL5 were higher after PEG stress induction,while Gu4CL2 was most significantly up-regulated after PEG stress.The expression of Gu4CL2 was similar to that of isoliquiritigenin.Bioinformatics analysis of Gu4CL gene showed that 11 genes had two conserved polypeptide motifs Box Ⅰ(SSGTTGLPKGV)and Box Ⅱ(GEICIRG),and 11 genes were distributed on 11 Scaffold fragments.Promoter cis-acting element analysis showed that Gu4CL2, Gu4CL4 and Gu4CL5 genes contained more abscisic acid and jasmonic acid response elements than other Gu4CL genes.Therefore,drought stress may induce the synthesis of jasmonic acid and abscisic acid,regulate the expression of Gu4CL2, Gu4CL4 and Gu4CL5 genes in licorice roots,and increase the accumulation of isoliquiritigenin.This study provides a research basis for optimizing the cultivation techniques of G.uralensis,improving the adaptability and quality of cultivated land in saline-alkali land,exploring the role of Gu4CL gene family in G.uralensis,and exploring the synthesis mechanism of isoliquiritigenin.

Key words: Liquorice, Gu4CL, Drought stress, Isoliquiritigenin, Bioinformatics analysis

赵斌, 姚华, 石娜娜, 高转转, 杨茂, 冯江华, 沈海涛. 甘草4CL基因家族鉴定及其在干旱胁迫下的表达分析[J]. 华北农学报, 2024, 39(S1): 1-10. doi: 10.7668/hbnxb.20194561.

ZHAO Bin, YAO Hua, SHI Nana, GAO Zhuanzhuan, YANG Mao, FENG Jianghua, SHEN Haitao. Identification of 4CL Gene Family in Glycyrrhiza uralensis and Its Expression under Drought Stress[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(S1): 1-10. doi: 10.7668/hbnxb.20194561.

http://www.hbnxb.net/CN/Y2024/V39/IS1/1

图/表 11

参考文献 28

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基因ID Gene ID	上游引物序列(5'-3') Upstream primer sequence(5'-3')	下游引物序列(5'-3') Downstream primer sequence(5'-3')
Glyur000051s00003417.1	AATTCGTCTTCACCTTCCTC	TGTGTGATGATGAGTTTCGT
Glyur001041s00027454.1	TTAGATGCTGCTGTCATACC	ACTCCAGGTACCAACTATCA
Glyur001000s00030117.1	CAATGTGGAGGCTAATACGA	GCAAGTACCCATCAGAATCA
Glyur000100s00008376	CTGATGCAGAGCTTCAAATCGAG	TTCGGAAGGAAGGTTGAGGTAAG

基因ID Gene ID	上游引物序列(5'-3') Upstream primer sequence(5'-3')	下游引物序列(5'-3') Downstream primer sequence(5'-3')
Glyur000051s00003417.1	AATTCGTCTTCACCTTCCTC	TGTGTGATGATGAGTTTCGT
Glyur001041s00027454.1	TTAGATGCTGCTGTCATACC	ACTCCAGGTACCAACTATCA
Glyur001000s00030117.1	CAATGTGGAGGCTAATACGA	GCAAGTACCCATCAGAATCA
Glyur000100s00008376	CTGATGCAGAGCTTCAAATCGAG	TTCGGAAGGAAGGTTGAGGTAAG

基因 Gene	地上部分表达量Aboveground part expression level					地下部分表达量Underground part expression level
基因 Gene	0 h	2 h	6 h	12 h	24 h	0 h	2 h	6 h	12 h	24 h
Gu4CL1	9.26±3.08b	8.59±2.55b	4.66±0.92b	13.72±4.02b	43.79±23.22a	0.82±0.74ab	0.85±0.32ab	0.52±0.38b	0.85±0.12ab	1.61±0.69a
Gu4CL2	36.58±13.22ab	46.83±12.04ab	23.09±13.52b	36.93±11.23ab	63.23±30.27a	6.41±4.38c	56.17±14.06a	19.89±8.03bc	34.03±3.09b	33.60±15.57b
Gu4CL3	36.64±10.85ab	30.14±8.68ab	41.21±3.50a	26.91±7.34bc	15.32±2.43c	21.27±5.66a	22.13±9.76a	20.57±3.15a	17.88±1.53ab	9.18±3.34b
Gu4CL4	6.87±3.91a	10.57±2.06a	5.32±2.31a	9.03±2.71a	8.79±7.11a	36.95±8.83b	124.52±66.94a	62.49±14.25ab	59.81±16.80ab	55.44±37.15ab
Gu4CL5	14.77±5.17c	38.94±14.93a	15.80±4.80c	5.61±1.88c	48.45±23.02a	2.53±1.15c	17.99±6.00a	4.93±3.35b	4.98±2.13b	2.42±1.24b
Gu4CL6	1.66±0.31b	2.63±0.90ab	2.19±0.81b	4.60±1.04a	3.71±2.19ab	5.15±0.17a	14.88±8.00a	9.93±2.72a	13.23±6.07a	6.76±5.10a
Gu4CL7	0.04±0.05a	0.00±0.00a	0.01±0.02a	0.04±0.04a	0.36±0.62a	0.93±0.29a	0.32±0.32a	0.52±0.32a	0.45±0.32a	0.52±0.37a
Gu4CL8	3.30±2.70a	3.47±1.69a	2.78±0.99a	7.81±4.93a	4.03±2.33a	0.38±0.32a	0.36±0.09a	0.34±0.18a	0.49±0.25a	0.86±0.55a
Gu4CL9	0.46±0.38ab	0.27±0.08b	0.35±0.22b	1.04±0.47a	0.80±0.30ab	0.40±0.40a	0.06±0.05a	0.19±0.13a	0.28±0.21a	0.20±0.22a
Gu4CL10	1.21±0.58ab	0.32±0.07b	1.40±1.52ab	5.35±5.30a	0.49±0.50b	0.25±0.22a	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b
Gu4CL11	2.73±0.76b	2.06±0.34b	2.54±1.57b	5.74±1.90a	2.58±1.91b	0.78±1.08a	0.39±0.54a	0.62±0.17a	0.08±0.09a	0.33±0.45a

基因 Gene	地上部分表达量Aboveground part expression level					地下部分表达量Underground part expression level
基因 Gene	0 h	2 h	6 h	12 h	24 h	0 h	2 h	6 h	12 h	24 h
Gu4CL1	9.26±3.08b	8.59±2.55b	4.66±0.92b	13.72±4.02b	43.79±23.22a	0.82±0.74ab	0.85±0.32ab	0.52±0.38b	0.85±0.12ab	1.61±0.69a
Gu4CL2	36.58±13.22ab	46.83±12.04ab	23.09±13.52b	36.93±11.23ab	63.23±30.27a	6.41±4.38c	56.17±14.06a	19.89±8.03bc	34.03±3.09b	33.60±15.57b
Gu4CL3	36.64±10.85ab	30.14±8.68ab	41.21±3.50a	26.91±7.34bc	15.32±2.43c	21.27±5.66a	22.13±9.76a	20.57±3.15a	17.88±1.53ab	9.18±3.34b
Gu4CL4	6.87±3.91a	10.57±2.06a	5.32±2.31a	9.03±2.71a	8.79±7.11a	36.95±8.83b	124.52±66.94a	62.49±14.25ab	59.81±16.80ab	55.44±37.15ab
Gu4CL5	14.77±5.17c	38.94±14.93a	15.80±4.80c	5.61±1.88c	48.45±23.02a	2.53±1.15c	17.99±6.00a	4.93±3.35b	4.98±2.13b	2.42±1.24b
Gu4CL6	1.66±0.31b	2.63±0.90ab	2.19±0.81b	4.60±1.04a	3.71±2.19ab	5.15±0.17a	14.88±8.00a	9.93±2.72a	13.23±6.07a	6.76±5.10a
Gu4CL7	0.04±0.05a	0.00±0.00a	0.01±0.02a	0.04±0.04a	0.36±0.62a	0.93±0.29a	0.32±0.32a	0.52±0.32a	0.45±0.32a	0.52±0.37a
Gu4CL8	3.30±2.70a	3.47±1.69a	2.78±0.99a	7.81±4.93a	4.03±2.33a	0.38±0.32a	0.36±0.09a	0.34±0.18a	0.49±0.25a	0.86±0.55a
Gu4CL9	0.46±0.38ab	0.27±0.08b	0.35±0.22b	1.04±0.47a	0.80±0.30ab	0.40±0.40a	0.06±0.05a	0.19±0.13a	0.28±0.21a	0.20±0.22a
Gu4CL10	1.21±0.58ab	0.32±0.07b	1.40±1.52ab	5.35±5.30a	0.49±0.50b	0.25±0.22a	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b
Gu4CL11	2.73±0.76b	2.06±0.34b	2.54±1.57b	5.74±1.90a	2.58±1.91b	0.78±1.08a	0.39±0.54a	0.62±0.17a	0.08±0.09a	0.33±0.45a

蛋白名称 Protein name	氨基酸数量/aa Number of amino acid	分子量/ku Molecular weight	等电点 pI	不稳定指数 Instability index	脂肪系数 Aliphatic index	亲疏水性 GRAVY	亚细胞定位 Subcellular location
Gu4CL1	556	60.865 43	6.91	38.65	101.78	0.201	质膜
Gu4CL2	588	64.016 18	5.60	38.64	95.14	-0.081	叶绿体
Gu4CL3	575	62.282 92	6.61	45.45	95.01	-0.015	质膜
Gu4CL4	525	56.811 20	5.48	41.18	93.81	0.063	质膜
Gu4CL5	526	56.783 97	6.11	33.75	96.62	-0.006	质膜
Gu4CL6	542	59.158 83	6.06	29.74	98.80	0.020	质膜
Gu4CL7	519	56.729 70	5.70	31.76	101.19	0.093	质膜
Gu4CL8	614	67.857 14	9.05	40.09	97.75	0.005	质膜
Gu4CL9	570	61.790 69	6.15	44.15	100.23	0.092	质膜
Gu4CL10	420	46.314 00	6.71	30.45	93.05	-0.011	质膜
Gu4CL11	553	60.075 32	6.37	45.16	98.92	0.052	内质网

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甘草4CL基因家族鉴定及其在干旱胁迫下的表达分析

Identification of 4CL Gene Family in Glycyrrhiza uralensis and Its Expression under Drought Stress

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蛋白名称 Protein name	α螺旋 Alpha helix	延伸链 Extended strand	β转角 Beta turn	无规则卷曲 Random coil
Gu4CL1	34.53	20.68	10.97	33.81
Gu4CL2	23.64	24.32	10.54	41.50
Gu4CL3	32.17	23.65	8.17	36.00
Gu4CL4	33.71	20.95	10.48	34.86
Gu4CL5	35.74	18.25	9.13	36.88
Gu4CL6	34.50	24.35	14.21	26.94
Gu4CL7	40.66	18.50	10.60	30.25
Gu4CL8	27.36	26.71	10.91	35.02
Gu4CL9	33.33	21.75	12.98	31.93
Gu4CL10	34.52	19.52	11.19	34.76
Gu4CL11	33.82	24.05	11.57	30.56

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甘草4CL基因家族鉴定及其在干旱胁迫下的表达分析

Identification of 4CL Gene Family in Glycyrrhiza uralensis and Its Expression under Drought Stress

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