库尔勒香梨PsLOX基因克隆及生物信息学分析

doi:10.7525/j.issn.1673-5102.2017.06.011

植物研究 ›› 2017, Vol. 37 ›› Issue (6): 876-886.doi: 10.7525/j.issn.1673-5102.2017.06.011

库尔勒香梨PsLOX基因克隆及生物信息学分析

张军¹, 刘旭新¹, 徐立生², 吐尔逊娜依¹, 李疆³

1. 新疆农业职业技术学院园林科技学院, 昌吉 831100;
2. 新疆喀什质量技术监督局, 喀什 844000;
3. 新疆农业大学林学与园艺学院, 乌鲁木齐 830052

收稿日期:2017-05-08 出版日期:2017-11-15 发布日期:2017-11-25
通讯作者: 李疆 E-mail:lijiangxj@163.com
作者简介:张军(1978-),男,副教授,博士研究生,主要从事果树种质资源方面的研究。
基金资助:
新疆维吾尔自治区自然科学基金项目（2016D01A053）

Cloning and Bioinformatics Analysis of PsLOX Gene in Pyrus sinkiangensis Yü. ‘Korla xiangli’

ZHANG Jun¹, LIU Xu-Xin¹, XU Li-Sheng², Tuerxunayi¹, LI Jiang³

1. Garden Science & Technology College, Xinjiang Agricultural Vocational Technical College, Changji 831100;
2. Xinjiang Kashgar Supervisory Bureau for Quality and Technology, Kashgar 844000;
3. College of Forestry and Horticulture, Xinjiang Agricultural University, Urumqi 830088

Received:2017-05-08 Online:2017-11-15 Published:2017-11-25
Supported by:
Xinjiang Uygur Autonomous Region Natural Science Foundation(2016D01A053)

摘要/Abstract

摘要： 克隆了库尔勒香梨（Pyrus sinkiangensis Yü）脂氧合酶（lipoxygenase，LOX）基因PsLOX，了解其在香梨果实不同发育时期的表达差异，为香梨果实香气代谢机理研究提供理论依据。以库尔勒香梨嫩叶及不同时期果实表皮为试材，利用两种不同的方法提取总RNA，通过RT-PCR技术得到目的基因PsLOX的cDNA序列，以生物信息学方法对其进行分析和功能预测。运用半定量RT-PCR （SqRT-PCR）技术，分析PsLOX基因在香梨嫩叶及果实生长发育及货架期的表达特性和差异。结果：试剂盒提取总RNA质量较高，PsLOX基因CDS序列为912 bp，编码303个氨基酸，属于脂氧合酶家族基因，与其他植物LOX基因编码的氨基酸序列有较高的同源性，与南果梨相似性最高，达到99%；PsLOX基因在香梨果实中发育过程中表达差异明显，即生长发育前期表达量很低，成熟至完熟时期表达量最高，然后开始减少。推测克隆获得PsLOX基因在香梨果实香气代谢过程中起到重要作用。

关键词: 库尔勒香梨, PsLOX基因, 半定量RT-PCR, 香气

Abstract: In order to provide a rationale and study for the aroma metabolism mechanism of pear fruit, which was PsLOX, the lipoxygenase gene of Korla pear(Pyrus sinkiangensis Yü. ‘Korla xiangli’) was cloned, the expression profiles of PsLOX were monitored in different periods of fruit development. As materials in leaves and fruit of Korla pear at different periods, total RNA was extracted by using the reagent kit, cDNA sequence of target gene PsLOX was obtained by RT-PCR. The function of PsLOX was predicted by bioinformatics, the expression characteristics and differences of this gene were analyzed using semi quantitative RT-PCR(SqRT-PCR) technology with leaves and peel of Korla pear in period of growth and shelf-life. The CDS sequence of PsLOX gene was 912 bp length, coded a predicted protein of 303 amino acids. By bioinformatics, PsLOX gene belongs to the lipoxygenase family. We compared with other plant LOX genes, the amino acid sequence had high homology, and the highest homology with Nanguo pear reached 99%. There was significant difference in the expression of PsLOX gene in development period of pear fruit, which was very low level in the expression quantity at growth development prophase of young fruit, and was the highest value in mature to ripe period, then expression quantity began to decrease. The PsLOX gene was homologously cloned, which were speculated to play a crucial role in aroma metabolism process of Korla pear fruit.

Key words: Pyrus sinkiangensis Yü.‘Korla xiangli’, PsLOX gene, semi quantitative RT-PCR, aroma

中图分类号:

S661.2

张军, 刘旭新, 徐立生, 吐尔逊娜依, 李疆. 库尔勒香梨PsLOX基因克隆及生物信息学分析[J]. 植物研究, 2017, 37(6): 876-886.

ZHANG Jun, LIU Xu-Xin, XU Li-Sheng, Tuerxunayi, LI Jiang. Cloning and Bioinformatics Analysis of PsLOX Gene in Pyrus sinkiangensis Yü. ‘Korla xiangli’[J]. Bulletin of Botanical Research, 2017, 37(6): 876-886.

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库尔勒香梨PsLOX基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of PsLOX Gene in Pyrus sinkiangensis Yü. ‘Korla xiangli’

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