ELISA法测定番茄茎叶中的褪黑素含量
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摘要
为了研究植物内源褪黑素与植物抗性的关系,以4叶1心的番茄幼苗为试验材料,采用酶联免疫吸附测定(ELISA)法测定正常番茄、鱼腥草提取液和灰霉菌侵染后的番茄茎叶中的褪黑素含量。结果表明:番茄叶片和茎的褪黑素含量分别为14.53、2.80 ng/g,二者之间差异极显著,叶片的褪黑素含量是茎的5.19倍;经鱼腥草提取液预处理和灰霉菌侵染后,番茄叶片的褪黑素含量显著增加,鱼腥草提取液预处理有助于减轻番茄叶片上的灰霉病发病症状,但并不依赖于叶片内源褪黑素含量的增加。可以检出的番茄茎叶中的褪黑素含量高于2.60 ng/g,就灵敏度而言,ELISA法完全可以用来检测番茄的组织器官的褪黑素含量。
In order to study the relationship between plant endogenous melatonin and the resistance of plants, tomato seedlings with four leaves were used as experimental materials. The contents of melatonin in stems and leaves of conventional tomato and tomato sprayed by Houttuynia cordata extracting solution and Botrytis cinerea were determined by ELISA. The results showed that the contents of melatonin in leaves and stems of tomato were significantly different. The content of melatonin in leaves was 5.19 times higher than that in stems, which were 14.53, 2.80 ng/g respectively, showing significant difference. After pretreatment with Houttuynia cordata extracting solution and Botrytis cinerea, the content of melatonin in tomato leaves increased significantly. Pretreatment with extract of Houttuynia cordata was helpful to alleviate the symptoms of gray mold in tomato stems and leaves. But it did not depend on the increase of endogenous melatonin content in leaves. In this experiment, the content of melatonin in tomato stems and leaves were higher than 2.60 ng/g. In terms of sensitivity, ELISA could be used to detect the content of melatonin in tomato leaves and stems.
In order to study the relationship between plant endogenous melatonin and the resistance of plants, tomato seedlings with four leaves were used as experimental materials. The contents of melatonin in stems and leaves of conventional tomato and tomato sprayed by Houttuynia cordata extracting solution and Botrytis cinerea were determined by ELISA. The results showed that the contents of melatonin in leaves and stems of tomato were significantly different. The content of melatonin in leaves was 5.19 times higher than that in stems, which were 14.53, 2.80 ng/g respectively, showing significant difference. After pretreatment with Houttuynia cordata extracting solution and Botrytis cinerea, the content of melatonin in tomato leaves increased significantly. Pretreatment with extract of Houttuynia cordata was helpful to alleviate the symptoms of gray mold in tomato stems and leaves. But it did not depend on the increase of endogenous melatonin content in leaves. In this experiment, the content of melatonin in tomato stems and leaves were higher than 2.60 ng/g. In terms of sensitivity, ELISA could be used to detect the content of melatonin in tomato leaves and stems.
引文
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[2]LERNER A B,CASE J D,HEINZELMAN R V,et al.Structure of melatonin[J].Journal of the American Chemical Society,1959,81(22):6084-6085.
[3]CHEN L,TAN D,REITER R J,et al.In vivo and in vitro effects of the pineal gland and melatonin on[Ca2++Mg2+]-dependent ATPase in cardiac sarcolemma[J].Journal of Pineal Research,2010,14(4):178-183.
[4]MANCHESTER L C,TAN D X,REITER R J,et al.High levels of melatonin in the seeds of edible plants:possible function in germ tissue protection[J].Life Sciences,2000,67(25):3023-3029.
[5]HARDELAND R,BACKHAUS C,FADAVI A,et al.N1-acetyl-5-methoxykynuramine contrasts with other tryptophan metabolites by a peculiar type of NO scavenging:cyclization to a cinnolinone prevents formation of unstable nitrosamines[J].Journal of Pineal Research,2010,43(1):104-105.
[6]ZHANG N,SUN Q,ZHANG H,et al.Roles of melatonin in abiotic stress resistance in plants[J].Journal of Experimental Botany,2015,66(3):647.
[7]刘圣明,海飞,车志平,等.4种杀菌剂及其复配剂对番茄灰霉病菌的毒力[J].植物保护,2017,43(2):230-234.
[8]刘敏.鱼腥草中生物碱类化学成分及其生物活性研究进展[J].天然产物研究与开发,2018,30(1):141-145.
[9]赵燕,王东华,赵曦阳.植物中褪黑素的研究进展[J].西北植物学报,2014,34(1):196-205.
[10]李秀,秦玉川,HUANG Z Y,等.蜜蜂褪黑素的测定方法[J].应用昆虫学报,2005,42(1):99-102.
[11]卢斯.鹅褪黑素受体Mel-1c基因的克隆和血清褪黑素含量的研究[D].武汉:华中农业大学,2013.
[12]孟涛,郑志竑,叶婧,等.成年大鼠脑透析液和血清中褪黑素含量的测定[J].福建医科大学学报,2015(2):67-71.
[13]RUI W,YANG X L,HUI X U,et al.Research progress of melatonin biosynthesis and metabolism in higher plants[J].Plant Physiology Journal,2016,52(5):615-627.
[14]欧科鹏,李尤简,郭若婷,等.绵羊甲状腺自主合成并分泌褪黑素的研究[J].畜牧兽医学报,2015,46(4):576-582.
[15]谭.褪黑素对应激所致肠道动力异常的作用及其机制研究[D].武汉:武汉大学,2011.
[16]AGUNBIADE F O,OLUOWOLABI B I,ADEBOWALEK O.Phytoremediation potential of Eichornia crassipes in metal-contaminated coastal water[J].Bioresource Technology,2009,100(19):4521-4526.
[17]HERNáNDEZ I G,GOMEZ F J,CERUTTI S,et al.Melatonin in Arabidopsis thaliana acts as plant growth regulator at low concentrations and preserves seed viability at high concentrations[J].Plant Physiology and Biochemistry,2015,94:191-196.
[18]MURCH S J,SAXENA P K.A melatonin-rich germplasm line of St John's wort(Hypericum perforatum L.)[J].Journal of Pineal Research,2010,41(3):284-287.
[19]TAN D X,HARDELAND R,MANCHESTER L C,et al.Functional roles of melatonin in plants,and perspectives in nutritional and agricultural science[J].Journal of Experimental Botany,2015,63(2):577.