| 大蒜几丁质酶基因AsCHI1的鉴定及其对盐胁迫的响应 |
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| 引用本文： | 张宇阳,周雪,刘灵艺,许吴俊,任旭琴,王广龙,熊爱生.大蒜几丁质酶基因AsCHI1的鉴定及其对盐胁迫的响应[J].中国农学通报,2022,38(5):23-29. |
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| 作者姓名： | 张宇阳 周雪 刘灵艺 许吴俊 任旭琴 王广龙 熊爱生 |
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| 作者单位： | 1.淮阴工学院生命科学与食品工程学院,江苏淮安 223003;2.淮阴工学院应用技术学院,江苏淮安 223003;3.南京农业大学园艺学院/作物遗传与种质创新国家重点实验室/农业农村部华东地区园艺作物生物学与种质创制重点实验室,南京 210095 |
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| 基金项目： | 青海省科技厅重点实验室项目“青海省蔬菜遗传与生理重点实验室”(2020-ZJ-Y02)； |
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| 摘 要： | 本研究旨在研究大蒜几丁质酶基因的序列特征及其对盐胁迫的响应,鉴定其抗逆功能。以大蒜品种‘苍山四六瓣’为研究材料,通过RT-PCR技术分离得到AsCHI1基因。采用BioXM 2.6、ProtParam、DNAMAN、SignalP 5.0、SOPMA、SWISS-MODEL、NCBI和MEGA5等软件分析核苷酸及其编码的氨基酸序列特征,利用荧光定量PCR技术检测其在不同组织以及盐胁迫下的表达特性。该基因开放阅读框全长933 bp,编码310个氨基酸,其编码的蛋白属于几丁质酶Class I类,同时隶属于GH19家族成员。氨基酸序列分析显示,AsCHI1在N端含有大多数几丁质酶具备的信号肽区域,在C端与其他物种CHI氨基酸序列一致性较高。在亲缘关系上与同属百合科的麝香百合LlCHI (QBZ68892.1)以及山茶科的茶CsCHI(XP_028075045.1)较为接近。实时荧光定量PCR技术分析表明,AsCHI1基因在大蒜根、蒜瓣和叶片中均有表达,但在根中表达最高。另外,盐胁迫能显著诱导AsCHI1基因在各组织内的表达。说明该基因可能在大蒜植株抵御盐胁迫的过程中发挥重要作用。
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| 关 键 词： | 大蒜 盐胁迫 AsCHI1基因 基因克隆 响应 |
| 收稿时间： | 2021-03-18 |
Garlic Chitinase Gene AsCHI1: Identification and Its Response to Salt Stress |
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| ZHANG Yuyang,ZHOU Xue,LIU Lingyi,XU Wujun,REN Xuqin,WANG Guanglong,XIONG Aisheng.Garlic Chitinase Gene AsCHI1: Identification and Its Response to Salt Stress[J].Chinese Agricultural Science Bulletin,2022,38(5):23-29. |
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| Authors: | ZHANG Yuyang ZHOU Xue LIU Lingyi XU Wujun REN Xuqin WANG Guanglong XIONG Aisheng |
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| Institution: | 1.School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huaian, Jiangsu 223003;2.Faculty of Applied Technology, Huaiyin Institute of Technology, Huaian, Jiangsu 223003;3.State Key Laboratory of Crop Genetics and Germplasm Enhancement/ Ministry of Agriculture Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China/ College of Horticulture, Nanjing Agricultural University, Nanjing 210095 |
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| Abstract: | The study aims to investigate the sequence characteristics of garlic chitinase gene and its response to salt stress, and to identify its function in resistance to stresses. The AsCHI1 gene was isolated by RT-PCR technology using garlic cultivar ‘Cangshan siliuban’ as the research material. BioXM 2.6, ProtParam, DNAMAN, SignalP 5.0, SOPMA, SWISS-MODEL, NCBI and MEGA5 were adopted to analyze the sequence characteristics of nucleotides and its encoded amino acids. Fluorescent quantitative PCR technology was introduced to detect its expression in different tissues and under salt stress. The open reading frame of the gene was 933 bp in length encoding 310 amino acids. The protein encoded by AsCHI1 gene belonged to chitinase Class I and GH19 family. Amino acid sequence analysis showed that AsCHI1 contained a signal peptide region at the N-terminus possessed by most chitinases, whereas the C-terminus was highly consistent with the amino acid sequences of CHI from other species. In terms of genetic relationship, it was relatively close to Lilium longiflorum LlCHI (QBZ68892.1) in the Liliaceae family and tea CsCHI (XP_028075045.1) in the Theaceae family. Real-time fluorescent quantitative PCR analysis showed that the AsCHI1 gene was expressed in garlic roots, garlic cloves and leaves, but the highest expression was observed in the roots. In addition, salt stress could significantly induce the expression of AsCHI1 gene in various tissues. This gene may play an important role in the resistance to salt stress in garlic plants. |
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| Keywords: | Allium sativum salt stress AsCHI1 gene gene cloning response |
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