Effect of Brassinolide on Root Architecture of Tobacco Seedlings and Regulation Mechanism

doi:10.15933/j.cnki.1004-3268.2022.08.005

Abstract

Abstract: In order to explore the regulation mechanism of brassinolide（BRs）on root architecture of tobacco seedlings，the differences of root architecture under different concentrations of exogenous EBR（2，4⁃epibrassinolide） and BRZ（brassinolide synthesis inhibitor） were investigated using the flue⁃cured tobacco Nongda 202.The changes of root tip cross⁃section of secondary lateral roots were observed by electron microscope，and the auxin content，GUS（β⁃glucosidase）staining and relative gene expression in lateral roots were determined to analyze the relationship between root architecture and auxin.The results showed that EBR could affected root length，root diameter and secondary lateral root density of tobacco seedlings.Lower concentration of EBR（0.1 nmol/L）increased the total root length by 29.0%，but higher EBR，such as 0.5，1，5，10 and 100 nmol/L declined the primary lateral root length by 4.6%，9.6%，18.5%，29.6% and 36.9% respectively.As the EBR concentration（0.1—10 nmol/L）increased，the density of secondary lateral roots increased，but the average root diameter gradually decreased.The average root diameter of tobacco increased significantly in BRZ treatment（100—500 nmol/L）.EBR affected the root diameter of tobacco seedlings mainly by regulating the number of tobacco lateral root cells and the diameter of column，the cross cutting diameters of secondary lateral root and column under 10 nmol/L EBR treatment were significantly reduced by 32.5% and 25.0% compared with CK（No EBR and BRZ）. The auxin content of lateral root decreased by 7.1% significantly under 10 nmol/L EBR treatment，but there was no change of lateral root auxin under 500 nmol/L BRZ treatment. EBR treatment increased the expression of PIN3aT，PIN3bS，PIN3bT and PIN11T in tobacco，which was considered to change the distribution of auxin in roots.Therefore，brassinolide can regulate the level of auxin in different parts of roots，and then affect the changes of tobacco root architecture.

Key words: Tobacco, Brassinolide, Root architecture, Lateral root density, Lateral root diameter, Auxin

摘要： 为探究油菜素内酯（BRs）对烟草幼苗根系构型的调控机制，选取烤烟品种农大202为试验材料，分析不同浓度外源EBR（2，4-表油菜素内酯）和BRZ（BRs合成抑制剂）处理下烟草幼苗根系构型的差异，通过电镜观察二级侧根根尖横截面变化，并观察烟草转基因DR5：：GUS 材料根系GUS（β-葡萄糖苷酸酶）染色情况，测定侧根中生长素含量和相关基因表达量，分析根系构型变化与生长素之间的关系。结果表明，EBR可以调节烟草幼苗根长、根系直径与二级侧根密度变化，较低浓度的EBR（0.1 nmol/L）使总根长增加，增幅达到29.0%，但在0.5、1、5、10、100 nmol/L EBR处理下，一级侧根长度与对照（不添加EBR和BRZ）相比分别降低4.6%、9.6%、18.5%、29.6%、36.9%；随着EBR浓度的增大（0.1~10 nmol/L），二级侧根密度不断增大，但根系平均直径逐渐减小，BRZ（100~500 nmol/L）处理后烟草根系平均直径显著增大；EBR主要通过调控烟草侧根细胞数目和中柱直径影响烟草幼苗根系直径大小，10 nmol/L EBR处理下烟草二级侧根横切直径较CK显著减少32.5%，中柱直径较CK显著减小25.0%；10 nmol/L EBR处理下，烟草侧根生长素含量较CK显著降低7.1%，500 nmol/L BRZ处理下，侧根生长素含量与CK相比无变化；EBR处理使烟草生长素运输基因家族中PIN3aT、PIN3bS、PIN3bT 和PIN11T 等基因上调，从而调控根中生长素的分布。可见，油菜素内酯可以调控根中不同部位生长素的水平进而影响烟草根系构型的变化。

关键词: 烟草, 油菜素内酯, 根系构型, 侧根密度, 侧根直径, 生长素

CLC Number:

S572

LIANG Dong, LIU Guangliang, SHI Yi, WANG Shusheng, WANG Chengdong, WANG Xiaolin. Effect of Brassinolide on Root Architecture of Tobacco Seedlings and Regulation Mechanism[J]. Journal of Henan Agricultural Sciences, 2022, 51(8): 37-44.

梁栋, 刘光亮, 石屹, 王树声, 王程栋, 王晓琳. 油菜素内酯对烟草幼苗根系构型的影响及调控机制[J]. 河南农业科学, 2022, 51(8): 37-44.

References

［1］CLOUSE S D，SASSE J M.Brassinosteroids：Essential regulators of plant growth and development［J］.Annual Review of Plant Biology，1998，49（1）：427.
［2］WANG X， KOTA U， HE K， et al.Sequential transphosphorylation of BRI1/BAK1 receptor kinase complex impacts early events in brassionosteroid signaling［J］.Developmental Cell，2008，15（2）：220.
［3］王喆，王璐，宋旭明，等.植物油菜素内酯信号通路与植物免疫相关研究进展［J］.安徽农业科学，2019，47（4）：26⁃29.
WANG Z，WANG L，SONG X M.Research progress of brassinolide signal pathway and plant immune［J］.Journal
of Anhui Agricultural Sciences，2018，47（4）：26⁃29.
［4］丁丹阳，张璐翔，朱智威，等.叶面喷施2，4-表油菜素内酯对烟草抗旱性的影响［J］.中国烟草科学，2018，39（4）：50⁃57.
DING D Y，ZHANG L X，ZHU Z W，et al.The effect of surface spraying 2，4⁃epibrassinolide on drought resistance of tobacco［J］.China Tobacco Science，2018，39（4）：50⁃57.
［5］赵雪松，王倩，闫青地，等.油菜素内酯对水稻根系发育的调控作用［J］.中国细胞生物学报，2016，38（10）：1191⁃1198.
ZHAO X S，WANG Q，YAN Q D，et al.The regulation of brassinolide on rice root development［J］.Chinese Journal
of Cell Biology，2016，38（10）：1191⁃1198.
［6］邓茜，张爱华，侯立江，等.油菜素内酯对小麦幼苗根系发育的影响［J］.麦类作物学报，2018，38（12）：1504⁃1511.
DENG Q，ZHANG A H，HOU L J，et al.The effect of brassinolide on the root development of wheat seedlings［J］.Journal of Triticeae Crops，2018，38 （12）：1504⁃1511.
［7］张歩阔.油菜素内酯对烤烟生长发育和产值的影响［J］.贵州农业科学，2001，29（4）：35⁃37.
ZHANG B K.The effect of brassinolide on the growth and production value of flue⁃cured tobacco［J］.Guizhou Agricultural Sciences，2001，29（4）：35⁃37.
［8］宋守晔，郭战伟，刘建康，等.外源植物激素在烟草生产中的应用效果及前景展望［J］.河南农业科学，2004（10）：29⁃32.
SONG S Y，GUO Z W，LIU J K，et al.Application effects and prospects of exogenous plant hormones in tobacco production［J］.Journal of Henan Agricultural Sciences，2004（10）：29⁃32.
［9］李小玲，华智锐，李静.油菜素内酯对秦岭高山杜鹃耐热性的影响［J］.河南农业科学，2017，46（8）：126⁃130.
LI X L，HUA Z R，LI J.Effect of brassinolide on tolerance of Rhodendron lapponicum in Qinling to heat stress［J］.Journal of Henan Agricultural Sciences，2017，46（8）：126⁃130.
［10］齐群钢，张六洲，韩锦峰，等.表油菜素内酯对烤烟植株根系活力和碳氮代谢影响的研究［J］.河南农业大学学报，1988（3）：315⁃324.
QI Q G，ZHANG L Z，HAN J F，et al.The effect of epibrassinolide on the root vigor and carbon and nitrogen metabolism of flue⁃cured tobacco plants［J］.Journal of Henan Agricultural University，1988（3）：315⁃324.
［11］韩锦峰，齐群纲，张秀梅，等.表油菜素内酯对烟株不同部位叶片的物质运输与积累的调配效应研究［J］.中国烟草，1988（1）：9⁃13.
HAN J F，QI Q G，ZHANG X M，et al.Study on the effect of epibrassinolide on the material transportation and accumulation of different parts of tobacco plant leaves［J］. Chinese Tobacco，1988（1）：9⁃13.
［12］刘芳，李永忠，文国松，等.不同植物生长调节剂对烤烟上部叶质量的影响［J］.广西农业科学，2005，36（4）：303⁃305.
LIU F，LI Y Z，WEN G S，et al.The effects of different plant growth regulators on the quality of flue⁃cured
tobacco upper leaves［J］.Guangxi Agricultural Sciences，2005，36（4）：303⁃305.

［13］赵阳.水分胁迫条件下生长素对烟草苗期发育的影响机理［D］.北京：中国农业科学院研究生院，2017.

ZHAO Y.The effect mechanism of auxin on tobacco seedling development under water stress［D］.Beijing：Graduate School of Chinese Academy of Agricultural Sciences，2017.
［14］贺凌霄，薛刚，孙聚涛，等.外源赤霉素对烤烟叶面腺毛和香气物质的影响［J］.河南农业科学，2021，50（1）：53⁃59.
HE L X，XUE G ，SUN J T，et al.Effects of exogenous gibberellin on flue⁃cured tobacco leaf trichome and
aromatic substances［J］.Journal of Henan Agricultural Sciences，2021，50（1）：53⁃59.
［15］陆彦，祁琰，张晓敏，等.高水分、富含淀粉植物组织的扫描电镜制备技术优化［J］.植物科学学报，2018，36（1）：119⁃126.
LU Y，QI Y，ZAHNG X M，et al.Optimization of scanning electron microscopy preparation technology for high⁃moisture，starch⁃rich plant tissues［J］.Acta Plant Sciences，2018，36（1）：119⁃126.
［16］龚晓崇，宋从凤，王鸣华，等.高效液相色谱法测定烟草和棉花中生长素含量［J］.江苏农业学报，2012，28（1）：225⁃227.
GONG X C，SONG C F，WANG M H，et al.Determination of auxin content in tobacco and cotton by high performance
liquid chromatography［J］.Jiangsu Journal of Agriculture，2012，28（1）：225⁃227.
［17］SHAHENSHAH，ISODA A. Effects of water stress on leaf temperature and chlorophyll fluorescence parameters in cotton and peanut［J］.Plant Production Science，2010，13（3）：269⁃278.
［18］SABATINI S，BEIS D，WOLKENFELT H，et al.An auxin⁃dependent distal organizer of pattern and polarity in the Arabidopsi root［J］.Cell，1999，99：463⁃472.
［19］简志英，陆嘉陵.油菜素内酯对小麦幼苗伸长的影响［J］.上海农业科技，1986（6）：38⁃39.
JIAN Z Y，LU J L.The effect of brassinolide on the elongation of wheat seedlings［J］.Shanghai Agricultural Science and Technology，1986（6）：38⁃39.
［20］骆炳山.油菜素内酯和赤霉素对小麦初期生长反应的比较［J］.华中农业大学学报，1986（1）：40⁃44.
LUO B S.Comparison of the response of brassinolide and gibberellin to the initial growth of wheat［J］.Journal of Huazhong Agricultural University，1986（1）：40⁃44.
［21］TAIZ L，ZEIGER E. 植物生理学［M］.宋纯鹏，王学路，周云，译.北京：科学出版社，2015：563.
TAIZ L，ZEIGER E. Plant physiology［M］.SONG C P，WANG X L，ZHOU Y，translating.Beijing：Science Press，2015：563.
［22］GUPTA A，SING H M，LAXMI A.Interaction between glucose and brassinosteroid during the regulation of lateral root development in Arabidopsis［J］.Plant Physiology，2015，168（1）：307.
［23］BEEMSTER G T S，BASKIN T I.Analysis of cell division and elongation underlying the developmental acceleration of root growth in Arabidopsis thaliana［J］.Plant Physiology，1998，116（4）：1515.
［24］侯立江.油菜素甾醇调控小麦幼苗根系发育的细胞学与分子基础［D］.杨凌：西北农林科技大学，2019.
HOU L J.The cytological and molecular basis of brassinosteroids regulating the root development of wheat seedlings ［D］.Yangling： Northwest A&F University，2019.
［25］BENKOVA E，MICHNIEWICZ M，SAUER M，et al.Local，efflux⁃dependent auxin gradients as a common module for plant organ formation［J］. Cell，2003，115：591⁃602.
［26］FRIM J，VIETEN A，SAUER M，et al.Efflux⁃dependent auxin gradients establish the apicalbasal axis of Arabidopsis［J］.Nature，2003，426：147⁃153.
［27］赵晨云，郭小丽，陈亚轲，等.生长素和独脚金内酯参与磷素调控水稻分蘖的机制初探［J］.河南农业科学，2022，51（1）：21⁃26.
ZHAO C Y，GUO X L，CHEN Y K，et al.Preliminary study on mechanism of auxin and strigolactone involved
in phosphate⁃modulated tillering of rice［J］.Journal of Henan Agricultural Sciences，2022，51（1）：21⁃26.
［28］REED R C，BRADY S R，MUDAY G K.Inhibition of auxin movement from the shoot into the root inhibits lateral root development in Arabidopsis［J］.Plant Physiology，1998，118：1369⁃1378.
［29］ULMASOV T，MURFETT J，HAGEN G，et al.AUX/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements［J］.Plant Cell，1997，9：1963⁃1971.
［30］DENG Q，WANG X，ZHANG D Z，et al.BRS1 function in facilitating lateral root emergence in Arabidopsis［J］.International Journal of Molecular Science，2017，18（7）：1549.
［31］张爱华.油菜素内酯在小麦初生根发育中的作用分析［D］.杨凌：西北农林科技大学，2018.
ZHANG A H.Analysis of the role of brassinolide in the development of wheat primary roots［D］.Yangling：Northwest A&F University，2018.
［32］BAO F，SHEN J J，BRADY S R，et al.Brassinosteroids interact with auxin to promote lateral root development in
Arabidopsis［J］. Plant Physiology，2004，134：1624⁃1631.
［33］邹雪，门淑珍.生长素的外输载体PIN蛋白家族研究进展［J］.中国细胞生物学学报，2013，35（5）：574⁃582.
ZOU X，MEN S Z.Research progress on the PIN protein family of auxin delivery carriers［J］.Chinese Journal of
Cell Biology，2013，35（5）：574⁃582.
［34］KEUSKAMP D H，POLLMANN S，VOESENEK L A，et al.Auxin transport through PIN⁃FORMED 3（PIN3）controls shade avoidance and fitness during competition［J］.Proc Natl Acad Sci USA，2010，107（52）：22740⁃22744.
［35］HARRISON B R，MASSON P H.ARL2，ARG1 and PIN3 define a gravity signal transduction pathway in root statocytes［J］.Plant J，2008，53（2）：380⁃392.

[1]	MA Yu, WANG Shuoli, ZHAO Xi, HOU Bingqing, XUE Zizhong, ZHANG Yang, XI Hongang, CHENG Yuyuan, SHI Xiangdong, XING Xuexia. Effects of Water⁃retaining Agent on the Physical and Chemical Properties of Topsoil and the Growth and Development of Tobacco Plants in Cigar Tobacco Field in the Semi⁃humid Area of Southwest Henan [J]. Journal of Henan Agricultural Sciences, 2024, 53(2): 46-55.
[2]	LI Fenfen, WANG Aixia, ZHAO Chen, BAI Tao, MAO Lan, ZHANG Baolin, LI Shengdong, SONG Zhaopeng, WANG Tao. Identification Method of Green Tobacco Leaf Positions Based on Hyperspectral Imaging Technology [J]. Journal of Henan Agricultural Sciences, 2024, 53(2): 144-151.
[3]	WU Yunjie, LIU Huan, WANG Ruizhen, WANG Haoyu, SUN Mi, ZHENG Cong, WU Jianping, ZHOU Jianjun, TIAN Binqiang, LI Chunguang. Research on the Fragmentation Rule during Cut Tobacco Processing [J]. Journal of Henan Agricultural Sciences, 2024, 53(2): 162-170.
[4]	LI Yaling, MA Rong, DING Wei, FENG Wenning, XU Bo, LIANG Miao, LIU Yubin, LIU Wei, WANG Bing, QIAO Xueyi. Study on Pyrolysis Behavior of Tobacco Cuticle Based on TG⁃FTIR⁃MS [J]. Journal of Henan Agricultural Sciences, 2024, 53(2): 171-180.
[5]	LI Jingjing, RONG Shibin, QIN Yanqing, ZHOU Jun, ZENG Dailong, ZHONG Qiu, ZHAO Yuanyuan, LIU Deshui, ZHANG Ruina, SHI Hongzhi. Analysis of Chemical Composition and Quality Difference in Different Sections of Cigar Filler and Wrapper Leaves [J]. Journal of Henan Agricultural Sciences, 2024, 53(1): 62-69.
[6]	REN Zhichao, LI Xiang, LI Xianfeng, WU Lili, WANG Jing, PENG Zhiliang, LIU Guoshun, YIN Quanyu. Effect of Biochar on Rhizosphere Soil Fungal Community Structure of Flue‑cured Tobacco Varieties with Different Resistance to Black Shank Disease [J]. Journal of Henan Agricultural Sciences, 2024, 53(1): 105-115.
[7]	PAN Yong, WU Qiaoyin, LI Linlin, SHI Youzhi, TAN Zaiyu, ZHANG Juan, WANG Jian. Application of Polyphenol Oxidase‑Producing Strains in Improving the Quality of Cigar Tobacco Leaves [J]. Journal of Henan Agricultural Sciences, 2023, 52(9): 173-180.
[8]	FU Bo, YANG Yongfeng, LIU Xiangzhen, ZHAO Sensen, LIU Maolin, JIA Guotao, NIU Yangyang, ZHANG Kunfang, YU Jianjun, PENG Guixin, JI Xiaoming. Study on Aroma Type Classification Model Based on Near Infrared Spectra of Flue⁃cured Tobacco [J]. Journal of Henan Agricultural Sciences, 2023, 52(7): 163-171.
[9]	HU Zhizhong, FENG Yanchuang, SONG Lingyong, HU Chao, WU Wentao, LÜ Yangbo, LIANG Miao, ZHANG Junsong. Study on the Melting and Pyrolysis Characteristics of 12 Aldehydes and Ketones Commonly Used in Heated Tobacco Product0 Flavoring [J]. Journal of Henan Agricultural Sciences, 2023, 52(7): 172-180.
[10]	ZHANG Liushuang, XIE Hui, YANG Jinqing, HAN Jiaqi, BAO Zhijuan. Effects of Tea Polyphenols on Physiological Indexes and Pb Accumulation in Tobacco under Pb Stress [J]. Journal of Henan Agricultural Sciences, 2023, 52(6): 34-40.
[11]	CHEN Jiading, HE Rong, XIAO Qingli, YUAN Ming, TAN Qizhong, PENG Kui, WEI Shuo, LI Shengchun. Influence of Temperature and Moisture Content on Thermophysical Properties of Tobacco Leaves and Establishment of Prediction Models [J]. Journal of Henan Agricultural Sciences, 2023, 52(6): 172-180.
[12]	FENG Kang, YANG Yuanyuan, LIU Qiong, CHENG Lan, HAO Haohao, WANG Pingping, JIN Weihuan, GUO Hongxiang. Cloning and Expression Analysis of NtCCX2 Gene in Tobacco [J]. Journal of Henan Agricultural Sciences, 2023, 52(5): 52-60.
[13]	ZHANG Xiaoli, YUAN Ye, WEI Huiqin, YANG Xinyu, LIU Yufeng, LIU Xin, GUAN Xining, SHEN Hongtao, DUAN Weidong, WANG Yanfang, LIU Ling. Quality Changes of Mudanjiang Tobacco Strips during Natural Aging in Warehouse of Henan Province [J]. Journal of Henan Agricultural Sciences, 2023, 52(4): 170-180.
[14]	HAN Wenlong, ZHENG Cong, CAO Kexin, ZHANG Luyang, XU Xiaowen, XIE Ke, ZU Qiongyao, XU Shixiao. Effect of NtCPS2 Gene on Development and Secretion Synthesis of Tobacco Gland Hair [J]. Journal of Henan Agricultural Sciences, 2023, 52(3): 48-55.
[15]	WANG Lin, WANG Yudong, ZHU Bao, LI Hu, ZHOU Hongshen, WANG Kaiyue, YAN Tiejun, ZHOU Ping, XIA Hai, ZHOU Min. Isolation and Identification of Dominant Mold and Its Moldy Volatile Metabolites in Tobacco Strips during Storage [J]. Journal of Henan Agricultural Sciences, 2023, 52(3): 101-108.