| 摘要: |
| 【目的】克隆中国水仙几丁质酶基因，分析其在水仙不同组织中的表达，为提高中国水仙的抗病性提供参考。【方法】利用RT-PCR技术克隆得到中国水仙几丁质酶基因，对其序列及编码氨基酸序列进行生物信息学分析，通过qRT-PCR分析该基因在感染基腐病、病毒病及褐斑病中国水仙根、叶中的表达。【结果】中国水仙几丁质酶基因的cDNA序列长768 bp，编码230个氨基酸，其编码蛋白的分子质量为25.4 ku，理论等电点为9.04，为不稳定蛋白，亲水性强，二级结构以α螺旋和无规则卷曲为主。该基因编码的蛋白与荷兰芹几丁质酶蛋白的亲缘关系最近，隶属于糖苷水解酶 19 家族，与溶菌酶相似超家族基因的保守结构域类似，推测其可能是兼具几丁质酶和溶菌酶的双功能酶。qRT-PCR分析表明，在水仙基腐病、病毒病及褐斑病发病过程中，水仙几丁质酶基因的表达量均显著提高。【结论】成功克隆得到中国水仙几丁质酶基因，推测该基因可能参与了水仙的抗病过程。 |
| 关键词: 中国水仙 几丁质酶基因 序列分析 抗病性 |
| DOI： |
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| 基金项目:福建省林业厅创新项目“福建优势特色花卉水仙引种与产业化关键技术研究”（闽林种站（2013）42号） |
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| Cloning and expression analysis of chitinase gene from Narcissus tazetta var.chinensis |
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LIN Xiao-hong,PAN He-li,PAN Dong-ming
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| Abstract: |
| 【Objective】This study aimed to clone chitinase gene of Narcissus tazetta var.chinensis and analyze its expressions in different tissues to improve the resistance to pathogenic microorganisms.【Method】The chitinase gene sequence was cloned by RT-PCR technique and analyzed by bioinformatics.Chitinase gene expression in roots and leaves with base rot,virus disease and brown blotch of Narcissus tazetta var.chinensis was analyzed by qRT-PCR.【Result】Chitinase cDNA sequence contained 768 bp and encoded 230 amino acids with molecular weight of 25.4 ku and theoretical isoelectric point of 9.04.The main secondary structure of the protein was α helix with random coils.The protein encoded by chitinase cDNA had the closest relationship with parsley,belonging to glycoside hydrolase family 19.It was similar to conserved domains of lysozyme similar gene superfamily,suggesting that it may be a bifunctional enzyme of chitinase and lysozyme.【Conclusion】Chitinase gene was successfully obtained and it possibly involved in disease resistant process of Narcissus tazetta var.chinensis. |
| Key words: Narcissus tazetta var.chinensis chitinase gene sequence analysis resistance to disease |
