Effects of Topping and Hormone on Nicotine in Upper Tobacco Leaves and Analysis of Source Sink Relationship

doi:10.15933/j.cnki.1004-3268.2024.07.006

Abstract

Abstract: In order to clarify the physiological mechanism of topping and hormone regulating nicotine synthesis and accumulation in flue‑cured tobacco，the variety K326 was used to study the effects of different topping measure and hormone interactions on traits of root and upper leaf，contents of nicotine and total nitrogen in upper leaves，key enzymes activity of nitrogen metabolism，gene expression of nicotine synthesis and transport in roots，and analyze the source and sink relationship of nicotine accumulation.The results showed that topping significantly increased nicotine content and accumulation in upper tobacco leaves，which were 1.41 and 1.85 times those without topping respectively.Spraying methyl jasmonate（MeJA） without topping significantly increased nicotine content by 14.60%，while spraying N‑1‑naphthylphthalamic acid（NPA） significantly decreased nicotine content by 10.01%.Application of salicylhydroxamic acid（SHAM）and naphthylacetic acid（NAA）after topping significantly reduced nicotine content，with 23.07% and 16.92% reduction ranges respectively.Jasmonic acid（JA）had limited effect on root development，and mainly promoted nicotine synthesis through directly regulating the expression of nicotine synthesis and transport genes.Auxin（IAA） mainly indirectly affected nicotine synthesis and accumulation by regulating roots development. Path analysis showed that the number of root tips had the largest direct effect on nicotine accumulation in root traits，and the root surface area had the largest overall effect on nicotine accumulation，which were the main‘source’indicators affecting nicotine synthesis.The direct effect of leaf area on nicotine accumulation was the largest among leaf traits，and the total effect of leaf dry weight on nicotine accumulation was the largest，which were the main‘sink’indicators affecting nicotine accumulation.By comprehensive analysis，IAA content increased after topping，and promoted the increase of root tips number，which increased the number of nicotine synthesis‘sources’. The activities of nitrate reductase（NR）and glutamine synthetase（GS）were enhanced after topping meanwhile，promoting nitrogen metabolism and nitrogen acquisition.JA increased in roots after topping，which directly enhanced the expression of PMT，QPT，A622 and NUP1 genes for nicotine synthesis and transport，and increased the intensity of nicotine synthesis.In addition，topping improved nicotine accumulation efficiency by promoting the increase of‘sink’capacity such as leaf area and leaf dry weight.

Key words: Flue?cured tobacco, Topping, Jasmonic acid, Auxin, Nicotine, Source sink relationship

摘要： 为明确打顶和激素调控烤烟烟碱合成与积累的生理机制，以烤烟品种K326为材料，研究了不同打顶措施与激素互作处理对烤烟根系、上部叶片性状，上部叶烟碱、总氮含量，根系氮代谢关键酶活性及烟碱合成与运输基因表达的影响，并分析烟碱积累的源库关系。结果表明，打顶显著提高上部烟叶烟碱含量与积累量，分别是不打顶处理的1.41倍、1.85倍；不打顶时喷施茉莉酸甲酯（MeJA）显著增加烟碱含量，增幅为14.60%，喷施N-1-氨甲酰苯甲酸萘酯（NPA）显著降低烟碱含量，降幅为10.01%；打顶后喷施水杨苷异羟肟酸（SHAM）、萘乙酸（NAA）显著降低烟碱含量，降幅分别为23.07%、16.92%。茉莉酸（JA）对根系发育的影响有限，主要通过直接调控烟碱合成与转运基因的表达促进烟碱合成，而生长素（IAA）主要通过调控根系发育间接影响烟碱合成与积累。通径分析显示，根系性状中根尖数对烟碱积累的直接作用最大，根表面积对烟碱积累的总作用最大，是影响烟碱合成的主要“源”指标；叶片性状中叶面积对烟碱积累的直接作用最大，叶干质量对烟碱积累的总作用最大，是影响烟碱积累的主要“库”指标。综合分析，打顶后根系IAA含量提高，促进根尖数的增加，即增加了烟碱合成“源”数量，同时增强硝酸还原酶（NR）、谷氨酰胺合成酶（GS）活性，促进氮代谢与氮吸收；打顶后根系JA含量增加，直接增强烟碱合成与转运基因PMT、QPT、A622 和NUP1 的表达，提高了烟碱合成强度；此外，打顶促进叶面积、叶干质量等“库”容的增大，提高了烟碱积累效率。

关键词: 烤烟, 打顶, 茉莉酸, 生长素, 烟碱, 源库关系

CLC Number:

S572

WANG Yuhua, WANG Dequan, WANG Yulin, ZHANG Yang, DONG Xiaowei, XIONG Ying, LIU Zhongqing, SUN Yanguo. Effects of Topping and Hormone on Nicotine in Upper Tobacco Leaves and Analysis of Source Sink Relationship[J]. Journal of Henan Agricultural Sciences, 2024, 53(7): 44-53.

王玉华, 王德权, 王玉林, 张杨, 董小卫, 熊莹, 刘中庆, 孙延国. 打顶和激素对上部烟叶烟碱的影响及源库关系分析[J]. 河南农业科学, 2024, 53(7): 44-53.

References

［1］林雨晟，金洪石，张皓楠，等.高效液相色谱同时检测不同类型烟叶中的多种生物碱含量［J］.中国烟草科学， 2021，42（3）：83-89.

LIN Y C，JIN H S，ZHANG H N，et al.Simultaneous determination of alkaloids in different types of tobacco by HPLC［J］.Chinese Tobacco Science，2021，42（3）：83-89.

［2］陈海生，王可，刘国顺.豫中烟区烟叶烟碱含量与土壤颗粒组成的空间变异性［J］.河南农业科学，2019，48 （1）：44-49.

CHEN H S，WANG K，LIU G S.Spatial variability of nicotine content in tobacco leaves and soil granule composition of tobacco planting fields in central Henan Province［J］.Journal of Henan Agricultural Sciences， 2019，48（1）：44-49.

［3］BECERRA G P，ROJAS-RODRÍGUEZ F，RAMÍREZ D， et al.Structural and functional computational analysis of nicotine analogs as potential neuroprotective compounds in Parkinson disease［J］.Computational Biology and Chemistry，2020，86：107266.

［4］刘春奎，刘艳芳，刘会杰，等.混合型卷烟烟气主要化学成分与感官质量的关系［J］.南方农业学报，2019，50 （6）：1319-1327.

LIU C K，LIU Y F，LIU H J，et al.Relationship between main smoke chemical components and sensory quality of blended type cigarette［J］.Journal of Southern Agriculture，2019，50（6）：1319-1327.

［5］李青山，彭玉龙，张玉琴，等.添加外源碳对植烟土壤供氮能力及烟叶质量的影响［J］.河南农业科学，2022，51 （10）：53-60.

LI Q S，PENG Y L，ZHANG Y Q，et al.Effects of exogenous carbon addition on nitrogen supply ability of tobacco planting soil and quality of tobacco leaf［J］.Journal of Henan Agricultural Sciences，2022，51（10）： 53-60.

［6］刘晨，韩玉环，杨龙，等.不同套袋免打顶处理对烤烟上部叶生长发育的影响［J］.山东农业科学，2022，54（7）60-65.

LIU C，HAN Y H，YANG L，et al. Effects of different bagging without topping techniques on growth and development of upper leaves of flue-cured tobacco［J］.Shandong Agricultural Sciences，2022，54（7）：60-65.

［7］邱尧，周冀衡，黄劭理，等.打顶后氮素供应和腋芽生长互作对烤烟烟碱积累的影响［J］.中国烟草科学，2016， 37（4）：19-23.

QIU Y，ZHOU J H，HUANG S L，et al.Interaction effects of nitrogen supply and axillary bud growth after topping on nicotine accumulation in flue-cured tobacco［J］.Chinese Tobacco Science，2016，37（4）：19-23.

［8］唐兴贵，赵羡波，班国相，等.顶端调控技术对烤烟K326 品质和部位结构比例的影响［J］.河南农业科学，2017， 46（8）：46-51.

TANG X G，ZHAO X B，BAN G X，et al.Effect of topping-control technology on quality and structure proportion of flue-cured tobacco K326［J］. Journal of Henan Agricultural Sciences，2017，46（8）：46-51.

［9］赵唯琦，桑应华，黄泽坤，等.不同打顶时期对烤烟及其上部烟叶产质量的影响［J］.山东农业科学，2019，51 （12）：20-25.

ZHAO W Q，SANG Y H，HUANG Z K，et al.Effects of different topping time on yield and quality of flue-cured tobacco and its upper leaves［J］.Shandong Agricultural Sciences，2019，51（12）：20-25.

［10］ZHANG M Y，ZHAO Y Y，YANG C T，et al.The combination of transcriptome and metabolome reveals the molecular mechanism by which topping and salicylic acid treatment affect the synthesis of alkaloids in Nicotiana tabacum L［J］.All Life，2022，15 （1）：147-159.

［11］QIN Y，BAI S L，LI W Z，et al.Transcriptome analysis reveals key genes involved in the regulation of nicotine biosynthesis at early time points after topping in tobacco（Nicotiana tabacum L.）［J］.BMC Plant Biology，2020，20（1）：30.

［12］李健忠，薛立新，朱金峰，等.打顶后喷施油菜素内酯和生长素对烤烟田间生长、碳氮代谢及烟叶品质的影响［J］.中国生态农业学报，2015，23（11）：1404-1412.

LI J Z，XUE L X，ZHU J F，et al.Effects of brassinolide and auxin on growth，carbon and nitrogen metabolism and tobacco quality of flue-cured tobacco leaves after topping ［J］.Chinese Journal of Eco-Agriculture，2015，23（11）：1404-1412.

［13］吐尔逊江·买买提，阿塔吾拉，郭江平，等.新疆滴灌棉花打顶后激素分布动态与响应特征研究［J］.新疆农业科学，2017，54（8）：1402-1403.

TURSUNJAN M，ATAWULLA，GUO J P，et al.Research on hormone dynamic distribution and responsive characteristics of drip irrigation cotton after topping in Xinjiang［J］.Xinjiang Agricultural Sciences， 2017，54（8）：1402-1403.

［14］SUI X，HE X，SONG Z，et al.The gene NtMYC2 a acts as a‘master switch’in the regulation of JA-induced nicotine accumulation in tobacco［J］.Plant Biology， 2021，23（2）：317-326.

［15］BIAN S Q，SUI X Y，WANG J H，et al.NtMYB305a binds to the jasmonate-responsive GAG region of NtPMT1a promoter to regulate nicotine biosynthesis［J］.Plant Physiology，2022，188（1）：151-166.

［16］李寒，蔡凯，高维常，等.基于水杨酸和茉莉酸拮抗作用的烟碱合成调节研究［J］.农业生物技术学报， 2023，31（8）：1595-1609.

LI H，CAI K，GAO W C，et al. Study on the regulation of nicotine synthesis based on the antagonism of jasmonic acid and salicylic acid［J］.Journal of Agricultural Biotechnology，2023，31（8）：1595-1609.

［17］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.

［18］韩雅婷，郭天骄，贾国涛，等.保水剂和抗蒸腾剂配施对土壤水分和云烟87化学成分、碳氮代谢的影响［J］.河南农业科学，2023，52（11）：57-65.

HAN Y T，GUO T J，JIA G T，et al.Effects of combined application of water-retaining agent and antitranspirant on soil moisture，chemical composition and carbon and nitrogen metabolism of Yunyan 87［J］.Journal of Henan Agricultural Sciences，2023，52（11）： 57-65.

［19］HAYASHI S，WATANABE M，KOBAYASHI M，et al.Genetic manipulation of transcriptional regulators alters nicotine biosynthesis in tobacco［J］.Plant and Cell Physiology，2020，61（6）：1041-1053.

［20］MO Z J，DUAN L L，PU Y Y，et al.Proteomics and co-expression network analysis reveal the importance of hub proteins and metabolic pathways in nicotine synthesis and accumulation in tobacco（Nicotiana tabacum L.）［J］.Frontiers in Plant Science，2022，13：860455.

［21］HU M Y，ZHANG H B，WANG B W，et al.Transcriptomic analysis provides insights into the auxin response factor 6-mediated repression of nicotine biosynthesis in tobacco（Nicotiana tabacum L.）［J］.Plant Molecular Biology，2021，107（1）：21-36.

［22］HENRY J B，VANN M C，LEWIS R S.Agronomic practices affecting nicotine concentration in flue-cured tobacco：A review［J］.Agronomy Journal，2019，111 （6）：3067-3075.

［23］LI R Y，ISLAM S U，WU Z J，et al.Bensulfuron-methyl treatment of soil affects the infestation of whitefly，aphid，and Tobacco mosaic virus on Nicotiana tabacum［J］.Frontiers in Plant Science，2016，7：1970.

［24］鲁雅雯，李斐，许晓敬，等.NtMYC1在打顶调控烟碱生物合成中的功能研究［J］.植物生理学报，2021，57 （1）：169-177.

LU Y W，LI F，XU X J，et al.Role of NtMYC1 in topping-induced nicotine biosynthesis in Nicotiana tabacum［J］. Plant Physiology Journal，2021，57（1）： 169-177.

［25］HAN G Z.Evolution of jasmonate biosynthesis and signaling mechanisms［J］.Journal of Experimental Botany，2017，68（6）：1323-1331.

［26］任永哲，徐艳花，李振声，等.拟南芥根系发育的分子机制研究进展［J］.西北植物学报，2011，31（7）：1497-1504.

REN Y Z，XU Y H，LI Z S，et al.Advances in the molecular mechanisms of root development in Arabidopsis thaliana［J］.Acta Botanica Boreali-Occidentalia Sinica，2011，31（7）：1497-1504.

［27］ASSENG S，KASSIE B T，LABRA M H，et al.Simulating the impact of source-sink manipulations in wheat［J］.Field Crops Research，2017，202：47-56.

［28］SONNEWALD U，FERNIE A R.Next-generation strategies for understanding and influencing source-sink relations in crop plants［J］.Current Opinion in Plant Biology，2018，43：63-70.

［29］李向东，张永丽.农学概论［M］.北京：中国农业出版社，2017.

LI X D，ZHANG Y L. Introduction to agronomy［M］.Beijing：China Agriculture Press，2017.

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