摘要
【目的】探讨梨黄化叶片复绿过程中铁氮协同机制。【方法】以‘砀山酥梨’为试材,生长期喷施FeSO4溶液,测定叶片总Chl、总Fe、Fe^2+和N含量,分析Fe、N代谢相关基因的表达量和N代谢相关酶活性。【结果】0.20%FeSO4处理复绿最明显,但叶色仍不如正常叶片浓绿。复绿叶片中总Chl、总Fe、Fe^2+和N含量显著高于黄化叶片。0.20%FeSO4处理复绿叶类囊体片层结构清晰,与正常叶片相比,仍有断裂片层。FeSO4处理黄化叶片8 d和12 d,FER1、FER2、FER3、FER4、FD1和FD2相对表达量显著高于黄化叶片。0.10%、0.15%和、0.20%FeSO4处理后4 d和8 d,IRT相对表达量显著高于黄化叶片。FeSO4处理显著提高了黄化叶片中NR、NiR、GLIE和NADH-GOGAT相对表达量及其酶活性。【结论】外源FeSO4促进了黄化叶片中铁的贮藏、还原与转运,也促进了氮的代谢。
【Objective】China has the largest pear cultivation area in the world.In 2017,the total cultivated area was about 960000 hm^2,and the total output was about 16.53 million tons,accounting for 69.1%and 68.4%of the total cultivated area and total output of the world,respectively.‘Dangshansuli’pear(Pyrus bretschneideri Rehd.)is the largest cultivated variety of pear tree in China,and it was extensively cultivated in the largest region of continuous cultivation,The Old Yellow River Course Region,in Anhui province.In recent years,the phenomenon of iron-deficiency chlorosis in the local areas was carried out.【Methods】The chlorotic and normal sample trees of‘Dangshansuli’pear were used for the experiment in the Horticulture Farm of Dangshan County,Anhui province in May 2020,and the following treatments were carried out by using a powered backpack sprayer during the growth period with one non-sprayed tree left as a buffer between the treated trees:(1)CK,normal leaf(N)and chlorotic leaf(C)treated with distilled water;(2)T1,0.05%FeSO4 solution;(3)T2,0.10%FeSO4 solution;(4)T3,0.15%FeSO4 solution;(5)T4,0.20%FeSO4 solution.Three replicate experiments were adopted for each treatment,and each single experimental unit consisted of three replicate treestrees.All trees were relatively consistent in tree shape and vigor,spaced at 4 m×6 m,with seamless crops between the rows,training with the open center system,and the orchard was managed as usual.The retrieved green status was observed on the 4th,8th and 12th day after treatments,the leaf samples were taken back to the laboratory and quickly frozen in liquid nitrogen and stored in the refrigerator at-80℃for further analysis.The content of total Chlorophyll in the leaf was determined by 80%acetone extraction;the contents of total Fe and Fe^2+were respectively measured by the inductively coupled plasma-atomic emission spectometry(ICP-AES)and the phenanthroline colorimetric method;the content of N was determined by micro-kjeldahl method;the chloroplast ultra structure was observed under electron microscope;the total RNA of leaf samples was extracted by the RNAprep Pure Plant Plus Kit(TIANGEN Biochemical Technology Co.Ltd.,Beijing,P.R.of China),the cDNA was synthesized using a fluorescence reverse transcription kit(TaKaRa Biotechnology Co.Ltd.,Dalian,P.R.of China)and the related genes expression of Fe and N metabolism was analyzed by qRT-PCR conducted on an AB StepOne RT-PCR cycler(Applied Biosystems,Foster City,CA,USA);and the related enzymes activity of N metabolism was detected by the ultraviolet spectrophotometry colorimetry.【Results】The more retrieved green spots was formed with 0.05%and 0.10%FeSO4,and the more retrieved green patches formed with 0.15%and 0.20%FeSO4 on the 12th day,and the retrieved green effect of the 0.20%FeSO4 was the most obvious,but the leave color was not as deeply green as the normal leaves.Compared to chlorotic leaves,FeSO4 treatment significantly increased the contents of total Chl,total Fe,Fe^2+and N in retrieved green leaves,but the contents of total Chl and N were still significantly lower than those in normal leavesleaves.On the 12th day after treatment with 0.20%FeSO4,the grana lamellae of thylakoid in the leaf were clearer than those of chlorotic leaves,but compared with the normal leaves there were still broken grana lamellae,the chlorotics leaves treated with FeSO4 solution could recover green color to a certain extent,but not completely.The relative expression of FER1,FER2,FER3 and FER4 on the 8th and 12th day after FeSO4 treatment was not only significantly higher than that in the chlorotic leaves,but also significantly higher than that in the normal leaves;the relative expression of FD1 and FD2 was also significantly higher than that in the chlorotic leaves.On the 4th and 8th day,the relative expression of IRT treated with 0.10%,0.15%and 0.20%FeSO4 was significantly higher than that in the chlorotic leaves.The relative expression of NR,NIR,GLIE and NADH-GOGAT genes with FeSO4 treatment increased significantly than that with chlorotic leaves,and the relative enzymes activities were also significantly higher.【Conclusion】Therefore,the chlorotic leaves showed a certain degree of retrieved green color,but still could not completely retrieve green color after application of FeSO4 in different concentrations.The exogenous FeSO4 not only promoted the storage,reduction and transport of iron in chlorotic leaves,but also promoted the reduction of nitric acid and nitrite,to synthesize the glutamic acid and glutamine.The results revealed that iron and nitrogen had a certain synergistic effect in retrieved green process of the chlorotic leaves in‘Dangshansuli’pear.
作者
贾兵
郭国凌
余桃
王友煜
叶振风
刘莉
刘普
衡伟
JIA Bing;GUO Guoling;YU Tao;WANG Youyu;YE Zhenfeng;LIU Li;LIU Pu;HENGWei(School of Horticulture,Anhui Agricultural University,Hefei 230036,Anhui,China)
出处
《果树学报》
CAS
CSCD
北大核心
2020年第11期1676-1686,共11页
Journal of Fruit Science
基金
梨树和桃树化肥减施增效关键技术与产品研发(2018YFD0201401)
国家现代农业产业技术体系建设专项(CARS-28-14)。
关键词
‘砀山酥梨’
复绿
铁
氮
增益作用
‘Dangshansuli’pear
Retrieved green
Iron
Nitrogen
Reinforced effect
作者简介
贾兵,男,副教授,在读博士研究生,研究方向为果树栽培生理。Tel:0551-65786607,E-mail:jb1977@ahau.edu.cn;通信作者:衡伟,Tel:15856949929,E-mail:hengwei@ahau.edu.cn。
