茶树氮吸收效率的早期鉴定技术研究

苏静静; 阮丽; 王丽鸳; 韦康; 吴立赟; 白培贤; 成浩

doi:10.13305/j.cnki.jts.2020.05.002

茶叶科学 >

2020 , Vol. 40 >Issue 5: 576 - 587

DOI: https://doi.org/10.13305/j.cnki.jts.2020.05.002

研究报告

茶树氮吸收效率的早期鉴定技术研究

苏静静 ,
阮丽 ,
王丽鸳 ,
韦康 ,
吴立赟 ,
白培贤 ,
成浩

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1.中国农业科学院茶叶研究所/国家茶树改良中心/农业部茶树生物学与资源利用重点实验室,浙江杭州 310008;
2.中国农业科学院研究生院,北京 100081

苏静静,女,硕士研究生,主要从事茶树耐贫瘠育种研究。

收稿日期: 2020-01-13

修回日期: 2020-03-17

网络出版日期: 2020-10-10

基金资助

中央级科研院所基本科研业务费专项（1610212018004）、浙江省农业（茶树）新品种选育重大科技专项（2016C02053-8）、现代农业产业技术体系建设专项资金资助（CARS-19）

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Early Identification of Nitrogen Absorption Efficiency in Tea Plants

SU Jingjing ,
RUAN Li ,
WANG Liyuan ,
WEI Kang ,
WU Liyun ,
BAI Peixian ,
CHENG Hao

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1. Tea Research Institute, Chinese Academy of Agricultural Sciences, National Center for Tea Improvement, Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Hangzhou 310008, China;
2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received date: 2020-01-13

Revised date: 2020-03-17

Online published: 2020-10-10

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摘要

氮是植物生长的重要营养元素,在茶树栽培过程中常需施用大量氮肥,不仅消耗大量的资源,施用不当还会造成一系列环境问题。培育氮肥高效利用的茶树品种是解决这一问题的重要途径,而建立快速筛选高效株系的早期鉴定方法对于缩短育种茶树育种年限具有重要意义。本研究分析龙井43（LJ43）和中茶108（ZC108）两个茶树品种在不同氮素水平下对氨态氮和硝态氮的吸收与利用数据,通过与¹⁵N同位素标记技术的比对,验证非损伤微测技术（NMT）和实时荧光定量（qRT-PCR）技术在早期鉴定茶树株系氮素吸收利用能力方面的可行性与实用性,以期建立茶树氮吸收效率的室内早期鉴定技术。试验结果表明,¹⁵N同位素标记技术的稳定性和可重复性分别为89.51%、99.26%,而NMT的稳定性、可重复性分别为95.22%、96.76%;两种方法测定结果均显示茶树具有明显的喜铵特性;硝酸根转运蛋白基因CsNRT3.2和CsNRT2.4在两个品种中均表现出诱导上调表达效应,相比中茶108,龙井43中CsNRT2.4和CsNRT3.2具有更高的表达量,表明LJ43对外界氮源的响应高于ZC108。综上所述,认为NMT技术可在短时间内处理并测得茶树的瞬时吸收速率,且试验材料损耗少,可以用于茶树氮瞬时吸收速率的早期鉴定;CsNRT2.4和CsNRT3.2的表达量一定程度上反映了茶树对硝态氮吸收的能力。本研究可为氮高效茶树品种的早期鉴定技术建立提供依据。

关键词： 茶树; 氮吸收速率; 非损伤微测技术; ¹⁵N同位素示踪; 实时荧光定量

本文引用格式

苏静静 , 阮丽 , 王丽鸳 , 韦康 , 吴立赟 , 白培贤 , 成浩 . 茶树氮吸收效率的早期鉴定技术研究[J]. 茶叶科学, 2020 , 40(5) : 576 -587 . DOI: 10.13305/j.cnki.jts.2020.05.002

Abstract

Nitrogen is one of the most important elements for tea plants, and it has been over supplied in tea gardens, which not only results in the waste of resources, but also causes a series of environmental problems. Therefore, to breed tea cultivars with high nitrogen efficiency, it is necessary to establish an early detection of nitrogen usage rates which could be developed for screening the strains of tea plants. This study analyzed the absorption and utilization of ammonia nitrogen and nitrate nitrogen in two tea cultivars Longjing 43 (LJ43) and Zhongcha 108 (ZC108) under different nitrogen levels. The ¹⁵N isotope labeling technology was applied to verify the feasibility and practicability of non-invasive micro-test technology (NMT) and real-time fluorescence quantitative (qRT-PCR) technology in early identification of nitrogen absorption and utilization abilities of tea lines. The purpose of this study was to establish an indoor early identification technology of nitrogen absorption efficiency for tea plants. The results show that the accuracy, stability and repeatability of ¹⁵N were 85.16%, 89.51% and 99.26% respectively. While the accuracy, stability and repeatability of NMT were 91.35%, 95.22% and 96.76% respectively. The two methods showed that tea plants had obvious ammonium preference. Moreover, the expressions of nitrate transporter genes CsNRT3.2 and CsNRT2.4 were up-regulated by nitrogen in both cultivars. Compared with ZC108, the expressions of CsNRT2.4 and CsNRT3.2 in LJ43 were higher, indicating that the response of LJ43 to external nitrogen applications was higher than that of ZC108. Finally, it was preliminarily summarized that the NMT technology could measure the instantaneous absorption rate of tea plants in a short time with little loss of experimental materials. It might also be applied for the early detection of the instantaneous nitrogen absorption rates of tea plants. Meanwhile, the results also show that the expressions of CsNRT2.4 and CsNRT3.2 could partly reflect the nitrogen absorption ability of tea plants. This study provided a basis to develop techniques in early identification of high nitrogen-efficient cultivars in tea plants.

Key words： tea plants; nitrogen uptake rate; non-invasive micro-test technology (NMT); ¹⁵N isotope-labeled; qRT-PCR

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