| There are many problems in the process of producing tobacco,especially its upper leaves,such as tobacco leaves inadequately open,too thick leaves,high nicotine content,internal coordination problems in chemical composition and so on.In order to solve these problems,the effects of spraying various exogenous substances(naphthylacetic acid,KH2PO4,"Yanboshi" foliage fertilizer)on growth development and physiological biochemical indexes of upper leaves of K326 were studied.Combined with tobacco proteomic analysis to study the impact of various exogenous substances on the yield and quality of upper leaves of flue-cured tobacco,and to explore the physiological and biochemical mechanisms.The main results are as follows:(1)The effects of exogenous substances on agronomic characters of upper leaves in flue-cured tobacco were more obvious as time goes on.30 days after topping,compared with the control treatment,the leaves area increased 8.9%in the treatment of spraying "Yanboshi" foliage fertilizer,the leaves area increased 6.1%in the treatment of spraying KH2PO4,the leaves area increased 3.0%in the treatment of spraying naphthylacetic acid."Yanboshi" foliage fertilizer can promote the increase of the width of the upper tobacco leaves,and the improvement of "Yanboshi" foliage fertilizer on the upper leaves area reached a significant level extremely.(2)Spraying "Yanboshi" foliage fertilizer significantly increased starch content and potassium content in upper leaves,while the total nicotine content decreased significantly and the content of reducing sugar improve to some extent;And spraying naphthylacetic acid decreased the content of total sugar,reducing sugar and starch of upper leaves;Spraying KH2PO4 significantly reduced reducing sugar content in upper leaves.30 days after topping,compared with the control treatment,the starch content in upper leaves increased by 20.1%,potassium content increased by 20.1%,the total nicotine content decreased by 5.6%in the treatment of spraying "Yanboshi" foliage fertilizer,the differences all have reached the extremely significant difference level.The ratio of reducing sugar and total sugar was 0.88 in this treatment;And reducing sugar content in upper leaves decreased by 21.4%,the starch content was reduced by 2.4%,potassium content increased by 7.6%,the total nicotine content decreased by 2.5%.The ratio of reducing sugar and total sugar was 0.77 in the treatment of spraying naphthylacetic acid;Starch content in upper leaves increased by 29.7%,potassium content increased by 13.9%,the total nicotine content increased by 6.5%.The ratio of reducing sugar and total sugar was 0.79 in the treatment of spraying KH2PO4.The growth rate in potassium content and the reduction ratio in total nicotine content in the treatment of spraying "Yanboshi" foliage fertilizer are larger than some other treatments,and the ratio of reducing sugar and total sugar was higher too.So chemical compositions of upper leaves were much more coordinated after spraying "Yanboshi" foliage fertilizer,which is possible to improve the quality of upper leaves of tobacco effectively.(3)As time goes on,each treatment of spraying exogenous substances compared with the control treatment,the chlorophyll a,chlorophyll b and carotenoid content of upper leaves overall showed a downward trend.The chlorophyll content gradually reduced in the process of tobacco leaves maturity.Each exogenous substances promoted the maturity of upper leaves of tobacoo.(4)Spraying "Yanboshi" foliage fertilizer significantly increased the activity of acid invertase(AI),sucrose synthase(SS)and sucrose phosphate synthase(SPS),and decreased the activity of nitrate reductase(NR),glutamine synthetase(GS).The growth of tobacco transition from nitrogen metabolism mainly to the metabolism of carbon accumulation mainly in later stage of growth.So the growth potential of tobacco was weaken,the normal maturity of tobacco was speeded up.Spraying KH2PO4 also significantly increased the activity of acid invertase(AI)and sucrose phosphate synthase(SPS),it instructed that KH2PO4 could also mature tobacco leaves in later stage of growth.Spraying naphthylacetic acid decreased the activity of acid invertase(AI)and sucrose synthase(SS),and increased the activity of nitrate reductase(NR).It instructed that growth hormone analogs promoted the growth of flue-cured tobacco.The growth potential of tobacco was strong,and nitrogen metabolic was exuberant.Tobacco is prone to the phenomenon of late-maturing.(5)49 differential protein spots were found among 4 protein profiles of leaves samples which derived from the growth period of 30 days after topping.Through LC-MS/MS detection and bioinformatics analysis,32 protein spots were identified:In the treatment of spraying "Yanboshi" foliage fertilizer.There are 5 defense-related proteins,3 energy-related proteins,2 proteins associated with photosynthesis and nutrition,2 proteins associated with protein synthesis,and 1 nucleic acid metabolism-related protein in up-regulated proteins.The expression of protein associated with photosynthesis and nutrition such as pyruvate orthophosphate dikinase,ribulose-1,5-bisphosphate carboxylase/oxygenase activase were raised,which promoted the CO2 assimilation accumulation and accelerated the growth of flue-cured tobacco.The leaves area of tobacco significantly increased and the enzyme activity associated with nitrogen metabolism decreased,then nitrogen metabolism of tobacco weakened,resulting in the decrease in nicotine content.While the larger leaf area,to some extent,also diluted the total nicotine content of tobacco leaves.Defense-related protein accounted for 38.5%of all the up-regulated proteins,and energy-related protein accounted for 23.1%.Under the high temperature,drought and other adverse environments in later stage of growth,tobacco may mainly increase the heat shock protein,ascorbate peroxidase,thioredoxin-like protein and so on to improve its adaptability.Thus tobacco have a high rate of metabolism,which is conducive to the absorption of potassium elements.Potassium in turn can increase the degree of hydration of the protoplasm,reduce its viscosity,enhance cell water retention capacity,improve its drought resistance.Under the combined effects of these factors,growth and development of the upper leaves of tobacco accelerated,yield increased and quality improved.There are 6 energy-related proteins,5 defense-related proteins,3 proteins associated with photosynthesis and nutrition,2 proteins associated with protein synthesis in up-regulated proteins under the treatment of spraying naphthylacetic acid.Defense-related protein and energy-related protein accounted for 68.8%of all the up-regulated proteins.In the role of naphthalene acetic acid,enzyme activity related carbon and nitrogen metabolism is generally low,while the expression of protein associated with energy metabolism such as Vacuolar H+-ATPase,formate dehydrogenase in mitochondria,ATP synthase CF1 a subunit,Phosphoglycerate kinase were raised,which provided more energy for the growth of tobacco.And the increase of the activity of nitrate reductase(NR)also shows that the nitrogen metabolism of tobacco is so strong that much energy is needed.Thus tobacco is prone to the phenomenon of late-maturing.In the treatment of spraying KH2PO4,there are 3 proteins associated with photosynthesis and nutrition,2 energy-related protein in up-regulated proteins.In the role of KH2PO4,tobacco enhanced its photosynthesis and energy metabolism.Carbon metabolism related enzyme activity increased significantly,which increased the starch and potassium content of leaves.The activity of enzyme related to Carbon metabolism was also significantly increased,so the the starch and potassium content of leaves increased.The activity of glutamine synthetase(GS)is high,which explained the rise of total nicotine content in tobacco leaves.In summary,compared with naphthylacetic acid and KH2PO4,spraying "Yanboshi" foliage fertilizer improving foliage area of upper leaves of tobacco to a greater extent.Tobacco achieved the purpose of increasing potassium reducing nicotine through increasing photosynthesis and nutrient metabolism and decreasing nitrogen metabolism.The chemical composition of tobacco was more coordinated.Moreover,tobacco increased its resistance to all kinds of adverse conditions on account of many defense-related proteins and energy-related protein up-regulated in the treatment of spraying "Yanboshi" foliage fertilizer.Thus,it will lay the foundation for stable yield and increasing income of flue-cured tobacco. |
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