Response of Root Development and Auxin in Tea Plants to Different Nitrogen Forms

doi:10.15933/j.cnki.1004-3268.2024.07.007

Abstract

Abstract: Tea plants（Camellia sinensis）prefer NH₄⁺ as the dominant nitrogen（N）source.Although it has been proven that the use of mixed nitrate（NO₃^-）and ammonium（NH₄⁺）as a N source can improve plant growth，whether genotypic difference in response to mixed nutrition exists in different tea plants remains obscure.Two tea cultivars（Camellia sinensis cv.Xinyang 10 and Zhongcha 108）were grown at three ratios of NH4+/NO₃^-（4∶0，3∶1 and 0∶4）to elucidate how the ratio of NH₄⁺/NO₃^- modulate the architecture of the root system by investigating root growth，N content and auxin content as well as the expression of related genes.Xinyang 10 had the highest root and shoot dry matter and root N content under mixed nutrition（3∶1）And Xinyang 10 had the longer adventitious roots under mixed nutrition relative to sole NH₄⁺，but no difference was observed in growth of lateral root between sole NH4+ andmixed nutrition treatments.IAA content was significantly increased in the roots of Xinyang 10，but significantly decreased in its leaves.Response of Zhongcha 108 to ratios of NH₄⁺/NO₃^- was different from Xinyang 10.Shoot biomass and N content of root and shoot in Zhongcha 108 were significantly reduced under mixed nutrition relative to sole NH₄⁺，while length of adventitious root increased，but not reaching statistical significance. Compared with sole NH₄⁺，mixed nutrition increased length of lateral root but decreased the number and density of lateral roots in Zhongcha 108.And the IAA content in roots of Zhongcha 108 was significantly reduced under mixed nutrition relative to sole NH₄⁺，but not in its leaves.The results of qRT‑PCR showed that auxin synthetic genes CsTSA，CsNIT1，transportation genes CsPIN3a，CsPIN3b and response genes CsARF3，CsIAA27 in roots of Xinyang 10 were significantly up‑regulated under mixed nutrition compared with sole NH₄⁺；however，auxin synthetic genes CsNIT1，CsNIT2，transportation CsABCB4 and response genes CsARF1，CsARF3，CsARF5，CsARF6 in roots of Zhongcha 108 were significantly down‑regulated under mixed nutrition compared with sole NH₄⁺. In conclusion，Xinyang 10 and Zhongcha 108 have genotypic differences when responding to N forms.The former is tolerant to NO₃^- and suitable for growth at the ratio of NH₄⁺/NO₃^-as 3∶1，while the latter is sensitive to NO₃^-and suitable for growth under sole NH₄⁺ nutrition. Under the ratio of NH4+/NO₃^- at 3∶1，auxin transport to roots of Xinyang 10 increases but to those of Zhongcha 108 decreases，which may be one of the physiological mechanisms conferring different response of different tea roots to N forms.

Key words: Camellia sinensis, Nitrogen form, NH₄⁺-N, NO₃^--N, Xinyang 10, Zhongcha 108, Roots, Auxin

摘要： 以茶树信阳10号和中茶108为试材，利用水培试验研究了不同氮形态［铵硝比（NH₄⁺∶NO₃^-）分别为4∶0、3∶1和0∶4］下茶树生物量、氮含量、根系发育、生长素含量及其相关基因的表达情况，旨在揭示不同氮形态下茶树根系的发育机制。结果表明，信阳10号在铵硝比3∶1下，根系和地上部干质量最高，根系氮含量最高。与单独供铵（4∶0）相比，信阳10号在铵硝比3∶1时，不定根显著伸长，侧根长和密度无显著变化，根系IAA含量显著增加，叶片IAA含量显著降低。与信阳10号不同，与单独供铵相比，中茶108在铵硝比3∶1下地上部干质量、根系和地上部氮含量显著减少，不定根长增加但差异不显著，侧根长显著伸长，侧根数目和侧根密度显著降低，IAA含量在根系中显著减少，叶片中无显著变化。qRT-PCR结果表明，铵硝比3∶1下，信阳10号根系生长素合成基因CsTSA、CsNIT1，运输基因CsPIN3a、CsPIN3b 和响应基因CsARF3、CsIAA27 表达显著上调；中茶108 根系生长素合成基因CsNIT1、CsNIT2，运输基因CsABCB4和响应基因CsARF1、CsARF3、CsARF5、CsARF6表达显著下调。综上所述，信阳10号和中茶108对不同氮素形态响应存在基因型差异，前者耐NO₃^-，适宜生长在铵硝比3∶1下，后者不耐NO₃^-，适宜生长在纯铵营养条件下；铵硝比3∶1下，生长素向信阳10号根系运输增加和向中茶108根系运输减少，可能是不同茶树根系对氮素形态响应不同的生理机制之一。

关键词: 茶树, 氮素形态, 铵态氮, 硝态氮, 信阳10号, 中茶108, 根系, 生长素

CLC Number:

S606⁺.2

HUANG Shuangjie, LI Mengzhen, LUO Jinlei, CHANG Yali, ZHANG Yali, GUO Guiyi. Response of Root Development and Auxin in Tea Plants to Different Nitrogen Forms[J]. Journal of Henan Agricultural Sciences, 2024, 53(7): 54-65.

黄双杰, 李梦真, 罗金蕾, 常亚丽, 张亚丽, 郭桂义. 茶树根系发育及生长素对不同氮形态的响应[J]. 河南农业科学, 2024, 53(7): 54-65.

References

［1］BLOOM A J.The increasing importance of distinguishing among plant nitrogen sources［J］.Current Opinion in Plant Biology，2015，25：10‑16.

［2］CHEN W，LUO J K，SHEN Q R.Effect of NH₄⁺‑N/NO₃^-‑N ratios on growth and some physiological parameters of Chinese cabbage cultivars［J］.Pedosphere 15（3）：310‑318.

［3］QIAN X Q，SHEN Q R，XU G H，et al.Nitrogen form effects on yield and nitrogen uptake of rice crop grown in aerobic soil［J］.Journal of Plant Nutrition，2004，27 （6）：1061‑1076.

［4］SONG W J，MAKEEN K，WANG D S，et al.Nitrate supply affects root growth differentially in two rice cultivars differing in nitrogen use efficiency［J］.Plant and Soil，2011，343（1）：357‑368.

［5］CAO W，TIBBITTS T W.Study of various NH₄⁺/NO₃^-mixtures for enhancing growth of potatoes［J］.Journal of Plant Nutrition，1993，16（9）：1691‑1704.

［6］徐子先，李银水，韩配配，等.不同铵硝配比对芝麻苗期光合荧光特性的影响［J］.河南农业科学，2017，46（9）：37‑44.
XU Z X，LI Y S，HAN P P，et al.Effect of different NH₄⁺‑N/NO₃^-‑N ratios on the photosynthetic and fluorescence characteristics of sesame seedlings［J］.Journal of Henan Agricultural Sciences，2017，46（9）：37‑44.
［7］陈浩婷，徐梦珠，李颖，等.不同铵硝配比下外源硅对黄瓜幼苗生长及抗氧化特性的影响［J］.山西农业科学，2022，50（10）：1429‑1437.
CHEN H T，XU M Z，LI Y，et al.Effects of exogenous silicon on the growth and antioxidant characteristics of cucumber seedlings with different ammonium and nitrate ratios［J］.Journal of Shanxi Agricultural Sciences，2022，50（10）：1429‑1437.

［8］邢瑶，马兴华. 氮素形态对烟苗根系生长及氮素利用的影响［J］.中国烟草学报，2016，22（4）：52‑61.
XING Y，MA X H.Effect of different nitrogen forms on tobacco seedling root growth and nitrogen utilization［J］.Acta Tabacaria Sinica，2016，22（4）：52‑61.

［9］宋文静，宋科，董建新，等.铵硝混合营养对烟草苗期氮代谢酶及内源生长素的影响［J］.江苏农业科学，2019，47（8）：100‑104.
SONG W J，SONG K，DONG J X，et al.Influences of ammonium and nitrate nutrition on nitrogen metabolism enzymes and endogenous auxin in tobacco seedlings［J］.Jiangsu Agricultural Sciences，2019，47（8）：100‑104.

［10］MENG L，DONG J X，WANG S S，et al.Differential responses of root growth to nutrition with different ammonium/nitrate ratios involve auxin distribution in two tobacco cultivars［J］.Journal of Integrative Agriculture，2019，18（12）：2703‑2715.

［11］康利允，李晓慧，高宁宁，等.不同铵硝配比对甜瓜叶片生理特性及产量、品质的影响［J］.果树学报，2021，38（5）：760‑770.
KANG L Y，LI X H，GAO N N，et al.Effects of different ammonium/nitrate ratios on leaf physiological characteristics，yield and quality in muskmelon［J］.Journal of Fruit Science，2021，38（5）：760‑770.

［12］屈亚潭，高欣，冯晓东.不同铵硝配比对番茄苗期生长和光合特性的影响［J］.延安大学学报（自然科学版），2021，40（3）：16‑20.
QU Y T，GAO X，FENG X D.Effects of different ammonium‑nitrate ratios on growth and photosynthesis of tomato seedlings［J］.Journal of Yan’an University（Natural Science Edition），2021，40（3）：16‑20.

［13］袁嫚嫚，邬刚，王家宝，等.不同铵硝配比对辣椒产量、养分积累和氮肥利用率的影响［J］.中国瓜菜，2022，35（2）：43‑48.
YUAN M M，WU G，WANG J B，et al.Yield，nutrient accumulation and nitrogen use efficiency of pepper under different ammonium/nitrate ratios［J］.China Cucurbits and Vegetables，2022，35（2）：43‑48.

［14］王岩，王永平，曾庆稳，等.不同铵硝配比供应下的辣椒根系形态、伤流组分和养分积累［J］.四川农业大学学报，2023，41（4）：602‑608.
WANG Y，WANG Y P，ZENG Q W，et al.Root morphology，bleeding components and nutrient accumulation of pepper under different NH₄⁺/NO₃⁻ ratio ［J］.Journal of Sichuan Agricultural University，2023，41（4）：602‑608.

［15］张家君，许珊珊，曹光球，等.不同氮形态对杉木叶绿素荧光参数和叶绿体超微结构的影响［J］.西北林学院学报，2020，35（2）：24‑31.
ZHANG J J，XU S S，CAO G Q，et al.Effects of nitrogen forms on the chlorophyll fluorescence parameters and chloroplast ultra‑structure of Cunninghamia lanceolata［J］.Journal of Northwest Forestry University，2020，35（2）：24‑31.

［16］王立冬，梁海燕，王燕茹，等.不同氮素形态与配比对杉木和木荷幼苗光合特性及生长的影响［J］.四川农业大学学报，2023，41（2）：217‑224.
WANG L D，LIANG H Y，WANG Y R，et al.Effects of different nitrogen forms and ratios on photosynthetic characteristics and growth in Cunninghamia lanceolata and Schima superba seedlings［J］.Journal of Sichuan Agricultural University，2023，41（2）：217‑224.

［17］MORITA A，OHTA M，YONEYAMA T. Uptake，transport and assimilation of 15N‑nitrate and 15N‑ammonium in tea（Camellia sinensis L.）plants［J］.Soil Science and Plant Nutrition，1998，44（4）：647‑654.

［18］RUAN J Y，ZHANG F S，WONG M H.Effect of nitrogen form and phosphorus source on the growth，nutrient uptake and rhizosphere soil property of Camellia sinensis L［J］.Plant and Soil，2000，223（1）：65‑73.

［19］RUAN J Y，GERENDÁS J，HÄRDTER R，et al.Effect of nitrogen form and root‑zone pH on growth and nitrogen uptake of tea（Camellia sinensis）plants［J］.Annals of Botany，2007，99（2）：301‑310.

［20］RUAN J Y，GERENDÁS J，HÄRDTER R，et al.Effect of root zone pH and form and concentration of nitrogen
on accumulation of quality‑related components in green tea［J］.Journal of the Science of Food and Agriculture，2007，87（8）：1505‑1516.

［21］YANG Y Y，LI X H，RATCLIFFE R G，et al.Characterization of ammonium and nitrate uptake and assimilation in roots of tea plants［J］.Russian Journal of Plant Physiology，2013，60（1）：91‑99.

［22］FAN K，FAN D M，DING Z T，et al.Cs‑miR156 is involved in the nitrogen form regulation of catechins
accumulation in tea plant（Camellia sinensis L.）［J］.Plant Physiology and Biochemistry，2015，97：350‑360.

［23］RUAN L，WEI K，WANG L Y，et al. Characteristics of NH₄⁺ and NO₃⁻ fluxes in tea（Camellia sinensis）roots measured by scanning ion‑selective electrode technique ［J］.Scientific Reports，2016，6：38370.

［24］TANG D D，LIU M Y，ZHANG Q F，et al.Preferential assimilation of NH₄⁺ over NO₃^- in tea plant associated
with genes involved in nitrogen transportation，utilization and catechins biosynthesis ［J］.Plant Science，2020，291：110369.

［25］杜旭华，彭方仁.无机氮素形态对茶树氮素吸收动力学特性及个体生长的影响［J］.作物学报，2010，36 （2）：327‑334.

DU X H，PENG F R.Effect of inorganic nitrogen forms on growth and kinetics of ammonium and nitrate uptake in Camellia sinensis L［J］.Acta Agronomica Sinica，2010，36（2）：327-334.

［26］胡国策，蒋家月，田坤红，等.氮素形态和水平对茶树生理特性的影响［J］.安徽农业大学学报，2018，45 （4）：588-593.

HU G C，JIANG J Y，TIAN K H，et al. Effects of nitrogen forms and nitrogen levels on the physiological characteristics of tea plants［J］. Journal of Anhui Agricultural University，2018，45（4）：588-593.

［27］方翔，胡国策，孙琪璐，等.氮素形态对茶树叶片品质及其氮代谢相关基因的影响［J］.西北农林科技大学学报（自然科学版），2020，48（2）：52-59.

FANG X，HU G C，SUN Q L，et al. Effects of nitrogen forms on tea quality and nitrogen metabolism related genes in tea leaves［J］. Journal of Northwest A & F University（Natural Science Edition），2020，48（2）： 52-59.

［28］YU P，LI X X，WHITE P J，et al. A large and deep root system underlies high nitrogen-use efficiency in maize production［J］. PLoS One，2015，10（5）： e0126293.

［29］陈静，马钦，崔文芳，等.氮高效玉米根系构型及吸收特性的研究进展［J］.东北农业科学，2022，47（3）： 55-58. CHEN J，MA Q，CUI W F，et al. Research progress on root structure and absorption characteristics of nitrogen efficient maize［J］.Journal of Northeast Agricultural Sciences，2022，47（3）：55-58.

［30］王新超，杨亚军，陈亮，等.茶树氮素利用效率相关生理生化指标初探［J］.作物学报，2005，31（7）： 926-931. WANG X C，YANG Y J，CHEN L，et al. Preliminary study on physiological and biochemical indices related to nitrogen use efficiency in tea plant［Camellia sinensis （L.）O.Kuntze］［J］.Acta Agronomica Sinica，2005，31 （7）：926-931.

［31］HU Q Q，SHU J Q，LI W M，et al. Role of auxin and nitrate signaling in the development of root system architecture［J］.Frontiers in Plant Science，2021，12： 690363.

［32］JIA Z T，GIEHL R F H，VON WIRÉN N. Local auxin biosynthesis acts downstream of brassinosteroids to trigger root foraging for nitrogen ［J］. Nature Communications，2021，12：5437.

［33］LIU Y，VON WIRÉN N. Ammonium as a signal for physiological and morphological responses in plants ［J］. Journal of Experimental Botany，2017，68（10）： 2581-2592.

［34］JIA Z T，VON WIRÉN N. Signaling pathways underlying nitrogen-dependent changes in root system architecture： From model to crop species［J］.Journal of Experimental Botany，2020，71（15）：4393-4404.

［35］ÖTVÖS K，MARCONI M，VEGA A，et al. Modulation of plant root growth by nitrogen source-defined regulation of polar auxin transport［J］. The EMBO Journal，2021，40（3）：e106862.

［36］MURATORE C，ESPEN L，PRINSI B. Nitrogen uptake in plants：The plasma membrane root transport systems from a physiological and proteomic perspective［J］. Plants，2021，10（4）：681.

［37］OLATUNJI D，GEELEN D，VERSTRAETEN I. Control of endogenous auxin levels in plant root development ［J］. International Journal of Molecular Sciences，2017， 18（12）：2587.

［38］王瑜.茶树叶片中生长素合成和信号转导的分子机制研究［D］.南京：南京农业大学，2020.

WANG Y. Molecular mechanisms of auxin biosynthesis and signal transduction in leaves of tea plant［D］.Nanjing：Nanjing Agricultural University，2020.

［39］GEISLER M M. A retro-perspective on auxin transport ［J］.Frontiers in Plant Science，2021，12：756968.

［40］GOMES G L B，SCORTECCI K C. Auxin and its role in plant development：Structure，signalling，regulation and response mechanisms［J］.Plant Biology，2021，23 （6）：894-904.

［41］HAMMES U Z，MURPHY A S，SCHWECHHEIMER C. Auxin transporters：A biochemical view［J］.Cold Spring Harbor Perspectives in Biology，2022，14（2）： a039875.

［42］HU S K，LIU X Q，XUAN W，et al. Genome-wide identification and characterization of PIN-FORMED （PIN）and PIN-LIKES（PILS） gene family reveals their role in adventitious root development in tea nodal cutting（Camellia sinensis）［J］.International Journal of Biological Macromolecules，2023，229：791-802.

［43］LIU Y，VON WIRÉN N. Integration of nutrient and water availabilities via auxin into the root developmental program［J］.Current Opinion in Plant Biology，2022，65：102117.

［44］杨亚军，梁月荣.中国无性系茶树品种志［M］.上海：上海科学技术出版社，2014：53-100.

YANG Y J，LIANG Y R. A record of clonal tea tree varieties in China［M］. Shanghai：Shanghai Science and Technology Publishers，2014：53-100.

［45］吴伯千，梁月荣，潘根生.水培和土培茶树的显微及超微结构比较［J］.浙江农业大学学报，1992，18（4）： 24-27. WU B Q，LIANG Y，PAN G S. Comparison study on super and micro structures of soil and solution culture tea plants ［J］. Journal of Zhejiang Agricultural University，1992，18（4）：24-27.

［46］孙虎威，王文亮，刘尚俊，等.低氮胁迫下水稻根系的发生及生长素的响应［J］.土壤学报，2014，51（5）： 1096-1102.

SUN H W，WANG W L，LIU S J，et al. Formation of rice root regulated by nitrogen deficiency［J］. Acta Pedologica Sinica，2014，51（5）：1096-1102.

［47］王学奎.植物生理生化实验原理和技术［M］.2版.北京：高等教育出版社，2006.

WANG X K. Principles and techniques of plant physiology and biochemistry experiment［M］. 2nd ed. Beijing：Higher Education Publishers，2006.

［48］黄双杰，曹梦珍，陈凌芝，等.氮素胁迫条件下茶树根系发育及生长素的响应［J］.江苏农业学报，2023，39 （3）：814-821.

HUANG S J，CAO M Z，CHEN L Z，et al. Auxin response and tea plant roots formation regulated by nitrogen stress［J］.Jiangsu Journal of Agricultural Sciences，2023，39（3）：814-821.

［49］史成颖.茶树幼根EST文库构建及茶氨酸代谢相关基因表达分析［D］.合肥：安徽农业大学，2011.

SHI C Y. Construction of an EST library of young roots of the tea plant（Camellia sinensis），and expression analysis of genes related to theanine synthesis［D］.Hefei：Anhui Agricultural University，2011.

［50］WANG B B，ZHU X L，GUO X L，et al. Nitrate modulates lateral root formation by regulating the auxin response and transport in rice［J］.Genes，2021， 12（6）：850.

［51］ASIM M，ULLAH Z，XU F Z，et al. Nitrate signaling， functions，and regulation of root system architecture： Insights from Arabidopsis thaliana［J］.Genes，2020，11 （6）：633.

［52］LUO L，ZHANG Y L，XU G H. How does nitrogen shape plant architecture？［J］Journal of Experimental Botany，2020，71（15）：4415-4427.

［53］ASIM M，ULLAH Z，OLUWASEUN A，et al.Signalling overlaps between nitrate and auxin in regulation of the root system architecture：Insights from the Arabidopsis thaliana［J］.International Journal of Molecular Sciences，2020，21（8）：2880.

[1]	SU Qiqian, AN Fuquan, YU Longfeng. Research Progress on Absorption of Germanium in Soils and Its Accumulation Mechanism in Camellia sinensis [J]. Journal of Henan Agricultural Sciences, 2024, 53(4): 1-8.
[2]	HE Zhanxue, ZHU Taifu, LI Xin, SU Xiaolan, LIU Huimin, WANG Lianchun. Analysis of Differences and Mechanism of Resistance to Canker among Different Kiwifruit Rootstock‑Scion Combinations [J]. Journal of Henan Agricultural Sciences, 2023, 52(1): 95-107.
[3]	DAI Zhuoyi, ZHANG Xi, XU Zhiqiang, YAO Yifan, XUE Gang. Effect of Grafting on Curing Characteristics of Flue‑cured Tobacco [J]. Journal of Henan Agricultural Sciences, 2021, 50(8): 44-50.