草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析

doi:10.11733/j.issn.1007-0435.2021.07.003

草地学报 ›› 2021, Vol. 29 ›› Issue (7): 1397-1405.DOI: 10.11733/j.issn.1007-0435.2021.07.003

草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析

金一锋^1,2, 陈阳¹, 齐欣¹, 高岩松¹, 金忠民¹, 赵清峰¹, 王琦¹

1. 齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006;
2. 抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江齐齐哈尔 161006

收稿日期:2020-08-18 修回日期:2020-11-19 出版日期:2021-07-15 发布日期:2021-07-31
通讯作者: 陈阳,E-mail:Chenyang8368215@126.com
作者简介:金一锋(1984-),男,黑龙江鹤岗人,讲师,硕士研究生,主要从事草坪草逆境分子生物学研究,E-mail:Jinyifeng8368215@126.com
基金资助:
国家自然科学基金（31501785）；黑龙江省普通高等学校青年创新人才培养计划项目（UNPYSCT-2018102）；黑龙江省省属高等学校基本科研业务费科研项目（135309365）；齐齐哈尔大学大学生创新创业项目（202010232161，202010232170，202010232802）资助

Cloning and Expression Analysis of NRT1/PTR FAMILY 8.3 Gene in Kentucky Bluegrass

JIN Yi-feng^1,2, CHEN Yang¹, QI Xin¹, GAO Yan-song¹, JIN Zhong-min¹, ZHAO Qing-feng¹, WANG Qi¹

1. College of Life Science and Agro-Forestry, Qiqihar Univesity, Qiqihar, Heilongjiang Province 161006, China;
2. Heilongjiang Province Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, Qiqihar, Heilongjiang Province 161006, China

Received:2020-08-18 Revised:2020-11-19 Online:2021-07-15 Published:2021-07-31

摘要/Abstract

摘要： 植物硝酸盐转运蛋白（Nitrate transporter，NRT）可有效转运NO₃^-，提升氮素利用效率。为了解析草地早熟禾（Poa pratensis）适应不同浓度及不同形态氮素、干旱胁迫机理，本研究以草地早熟禾为试材，对NRT1/PTR FAMILY 8.3基因进行克隆、生物信息学分析，并分析了不同浓度及不同形态氮素以及干旱下该基因表达情况。研究结果：草地早熟禾NRT1/PTR FAMILY 8.3基因包含典型的主要协同转运蛋白超家族（Major facilitator superfamily，MFS）结构域，与二穗短柄草（Brachypodium distachyon）高度同源；荧光定量PCR分析表明，该基因根部、叶部的表达量显著高于茎部；氮浓度为7.5 mM时，不同形态氮素诱导下，NaNO₃处理组有利于该基因表达；无氮、高氮组（0，15 mM NaNO₃）基因的表达量显著高于适氮组（7.5 mM NaNO₃）；干旱胁迫可促进其表达。研究结果为探究NRT1/PTR FAMILY 8.3在不同氮素环境中的调节机制，培育氮素利用率高的优质草种提供理论基础。

关键词: 草地早熟禾, NRT1/PTR FAMILY 8.3, 基因克隆, 表达分析

Abstract: As the main part of urban landscaping,lawn plays an important role in improving the ecological environment. Poa pratensis L. is a cool-season turfgrass with high quality. Poa pratensis L. has a high demand for nitrogen nutrients. Nitrate transporter (NRT) in plants can effectively transport NO₃^- and improve nitrogen-use efficiency. In this study,the NRT1/PTR FAMILY 8.3 gene of Midnight II was cloned and analyzed its bioinformation and expression under different nitrogen concentrations,nitrogen forms and drought treatments. The main results showed that the NRT1/PTR FAMILY 8.3 gene was cloned from Poa pratensis L.,which belonged to the major facilitator superfamily (MFS) and was highly homologous to the NRT1/PTR FAMILY 8.3 gene from Brachypodium distachyon. qRT-PCR results showed that the NRT1/PTR FAMILY 8.3 gene was highly expressed in roots and leaves of Poa pratensis L. After induction with different nitrogen forms at the same concentration,the expression of NRT1/PTR FAMILY 8.3 gene in the NaNO₃ treatment group was the highest. The expression of NRT1/PTR FAMILY 8.3 gene under nitrogen starvation and high-concentration nitrate nitrogen (0,15 mM NaNO₃) was higher compared with that under suitable nitrogen (7.5 mM NaNO₃). Drought stress promoted the expression of NRT1/PTR FAMILY 8.3 gene to a certain extent. The cloning and expression analysis results of Poa pratensis L. NRT1/PTR FAMILY 8.3 provided a theoretical basis for further exploring its regulatory mechanism in different nitrogen environments,determining its affinity to nitrate nitrogen,and cultivating high-quality turfgrass varieties with high nitrogen-use efficiency.

Key words: Kentucky bluegrass, NRT1/PTR FAMILY 8.3 gene, Gene Cloning, Expression analysis

中图分类号:

Q943.2

金一锋, 陈阳, 齐欣, 高岩松, 金忠民, 赵清峰, 王琦. 草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析[J]. 草地学报, 2021, 29(7): 1397-1405.

JIN Yi-feng, CHEN Yang, QI Xin, GAO Yan-song, JIN Zhong-min, ZHAO Qing-feng, WANG Qi. Cloning and Expression Analysis of NRT1/PTR FAMILY 8.3 Gene in Kentucky Bluegrass[J]. Acta Agrestia Sinica, 2021, 29(7): 1397-1405.

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草地早熟禾NRT1/PTR FAMILY 8.3基因的克隆及表达分析

Cloning and Expression Analysis of NRT1/PTR FAMILY 8.3 Gene in Kentucky Bluegrass

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