CN105900989B - 褪黑素在提高蔬菜硫素吸收中的应用 - Google Patents
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- A01N43/36—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings
- A01N43/38—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings condensed with carbocyclic rings
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Abstract
本发明公开了一种褪黑素在提高蔬菜硫素吸收中的应用,具体地,所述应用包括如下步骤:(1)将褪黑素与附加剂混合后制成褪黑素水剂;(2)将所述褪黑素水剂喷施于缺硫条件的目标植物叶面或投加于目标植物生长的缺硫营养液或缺硫土壤中。本发明的研究表明外源补充褪黑素后能够提高植物对硫的吸收,同时可以增强硫代谢的关键酶活性,促进含硫化合物的合成,进而提高植物对逆境的耐性。尤其是在土壤缺硫情况下,褪黑素的效果更加明显。
Description
技术领域
本发明涉及褪黑素的应用技术领域,具体涉及将褪黑素应用于提高蔬菜硫素吸收。
背景技术
硫是植物所需的大量营养元素。在植物的新陈代谢过程中,硫参与了植物的叶绿素和酶的合成,是氨基酸、蛋白质的重要结构组成成分。金属硫蛋白含有大量巯基,这一特殊结构赋予了其螯合重金属离子及重金属解毒的功能。而其它的含硫化合物如谷胱甘肽更是参与植物细胞的其它抗胁迫反应中。因此硫的吸收运输和同化代谢,以及一系列具有重要生物学功能的含硫化合物的合成对植物的生长发育、耐逆、抗病虫害,产量与品质等方面都至关重要。
植物硫营养主要来源于土壤,硫酸盐是土壤中硫的主要存在形式,植物根系以主动的方式吸收土壤中的SO4 2-,经一系列的同化反应后,生成半胱氨酸(Cys),随后,以此为前体,植物可以合成各种代谢产物,参与植物耐逆境胁迫和农作物的产量品质调控中。在大气污染比较严重的地区,植物也可以从污染空气中吸收到少量的SO2和H2S。当前,世界各地土壤供硫现状比较严峻,再加上对工业含硫废气排放的控制以及长期施用不含硫或含硫少的化肥,作物缺硫的问题已经愈发严重。因此增强植物对硫元素的利用,除施用含硫化肥提供充足的硫营养源外,在大范围的低硫和缺硫情况下,更重要的是注重提高植物对硫的吸收和利用效率。
褪黑素(melatonin)主要是由哺乳动物和人类的松果体产生的一种结构简单、低分子量的吲哚类激素,在植物中同样广泛存在,虽然含量极少(通常以pg或ng水平存在),却在生理调节、增加植物抗逆性方面起着非常重要的作用,是一种光谱的抗氧化剂。
发明内容
本发明提供一种褪黑素在提高蔬菜硫素吸收中的应用,研究表明外源补充褪黑素后能够提高植物对硫的吸收,同时可以增强硫代谢的关键酶活性,促进含硫化合物的合成,进而提高植物对逆境的耐性。尤其是在土壤缺硫情况下,褪黑素的效果更加明显。
一种褪黑素在提高蔬菜硫素吸收中的应用。
具体地,所述应用包括如下步骤:
(1)将褪黑素与附加剂混合后制成褪黑素水剂;
(2)将所述褪黑素水剂喷施于缺硫条件的目标植物叶面或投加于目标植物生长的缺硫营养液或缺硫土壤中。
本发明的褪黑素制剂,主要成分为褪黑素(melatonin,MT),又名美拉酮宁、松果腺素,分子式为C13N2H16O2,化学名称为N-乙酰基-5-甲氧基色胺(N-acetyl-5-methoxytryptamine)(结构如图1所示),不溶于水,溶于丙酮、乙醇、乙醚等有机溶剂,对光线敏感,受热易破坏,主要是由哺乳动物和人类的松果体产生的一种胺类激素,目前发现褪黑素同样广泛存在于各类高等植物中,但含量极少。
外源补充或内源增加褪黑素能够提高植物对多种逆境尤其是镉胁迫的抗性,而含硫化合物在植物解毒中极为重要,本发明通过大量的时间探索证明了褪黑素和植物硫的吸收同化之间存在着一定的关系。
所述附加剂为常规附加剂,例如,甲醇、乙醇和水。
为减少有机溶剂对植物的毒害作用,优选地,将褪黑素溶解于少量的甲醇或乙醇后再稀释至所需浓度。
为减少杂质的影响,优选地,将褪黑素溶于去离子水中制成水剂。
为了使该制剂在水溶液中均匀分布并均匀喷洒于植物叶面上,优选地,所述水剂中还添加一定比例的表面活性剂。农用有机硅的表面活性效果最好,实际使用时优选农用有机硅,添加量为褪黑素水剂质量的万分之二。
优选地,所述目标植物为番茄。
进一步优选地,步骤(2)中将番茄种植于缺硫营养液(标准硫1/10th)中,然后置于人工培养箱;待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔4~6天喷施一次褪黑素水剂,喷施总次数为2~4次。
更进一步优选地,待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔5天喷施一次褪黑素水剂,喷施总次数为3次。
进一步优选地,喷施时间为傍晚,喷适量以所有叶片湿润为准。
所述褪黑素水剂中褪黑素的浓度为25~500μM;进一步优选为50~150μM;最优选为100μM。
本发明中进一步优选地,待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔5天喷施一次褪黑素水剂,喷施总次数为3次;所述褪黑素水剂中褪黑素的浓度为50~150μM。
最优选地,待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔5天喷施一次褪黑素水剂,喷施总次数为3次;所述褪黑素水剂中褪黑素的浓度为100μM。与施加硫肥相比,本发明具有如下有益效果:
(1)方便安全。褪黑素制剂可通过将褪黑素与常规的附加剂混合后制成水剂得到,方便易得。褪黑素本身不会对土壤结构产生影响,对环境也不会产生任何污染,非常安全。
(2)提高硫的利用率。以适当浓度通过叶面喷施或者直接投加应用于营养液中或土壤中,可有效提高植物对硫的吸收、同化以及含硫化合物的合成。与单纯的补充硫肥相比,从本质上提高植物对硫的利用。
(3)好处多。除能够提高植物对硫的吸收外,本发明中褪黑素制剂本身就是一种光谱的抗氧化剂,外源补充后能够提高植物对所处不利环境的抗性,增加产量。
附图说明
图1为褪黑素分子结构。
图2为缺硫条件下和外源补充褪黑素后番茄植株的表型图。
图3为缺硫条件下和外源补充褪黑素后番茄光合速率对比图。
图4为缺硫条件下和外源补充褪黑素后番茄叶绿素含量对比图。
图5为缺硫条件下和外源补充褪黑素后番茄PSII最大光化学效率对比图。
图6为缺硫条件下和外源补充褪黑素后番茄生物量对比图。
图7~11分别为缺硫条件下和外源施加褪黑素水剂后,番茄根部和叶片中褪黑素含量及合成基因表达、硫含量以及硫转运蛋白基因的表达。
图12~13分别为缺硫条件下和外源施加褪黑素水剂后,番茄根部和叶片中硫同化代谢相关酶的活性和编码基因的表达对比图。
图14~16分别为缺硫条件下和外源施加褪黑素水剂后,番茄体内含硫化合物半胱氨酸、谷氨酰半胱氨酸以及谷胱甘肽(Cys、γ-EC、GSH)的含量对比图。
具体实施方式
实施例1
⑴植物材料
番茄(SolanumlycopersicumL.cv.Ailsa Craig)种子播种于蛭石和珍珠岩(v:v=50:50)的混合基质中,发芽后,浇灌1/2霍格兰营养液。一周后,一半植株转移至缺硫(标准硫的1/10th)营养液中,另一半作为正常对照。番茄幼苗置于人工气候培养箱培养,培养条件如下:昼夜温度23/20℃,相对湿度80%,光强800μmol m-2S-1以及光周期白天/黑暗为14h/10h。
(2)褪黑素制剂制备
参照公开号为CN104823738A的发明申请,研究了(25~500μM)不同浓度褪黑素制剂在植物硫素吸收中的影响,结果表明50~150μM之间的效果更好,100μM的褪黑素制剂效果最好,以下实验中以100μM的褪黑素制剂为例。褪黑素制剂组成如下:98%褪黑素(阿拉丁,上海晶纯生化科技股份有限公司)、乙醇、去离子水。先用少量的乙醇(250μl-500μl)溶解褪黑素,再加水稀释至5mM的母液。母液-20℃或4℃保存,使用时稀释至100μM,并加入体积比为万分之二的表面活性剂,实际应用中使用农用有机硅。所有操作尽量避光进行。
(3)喷施
将番茄种植于缺硫营养液(标准硫1/10th)中,然后置于人工培养箱;待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔4~6天喷施一次褪黑素水剂,喷施总次数为2~4次,优选每隔5天喷施一次褪黑素水剂,喷施总次数为3次。
(4)硫含量测定
叶片或根样,烘干后,称取0.3g,过筛后加高氯酸和硝酸(v:v=1:3)的混合液,180℃消煮至澄清。加2ml稀硝酸(50%),用去离子水定容样品至50ml。然后用比浊法测定硫含量。
(5)结果分析
如图2~6所示,缺硫引发番茄生长受阻,叶片小并且黄化(图2),光饱和CO2最大利用效率(Asat,图3)、叶绿素含量(图4)、PSII最大光化学效率(Fv/Fm,图5)以及植物生长量(图6)与对照植株相比分别降低了41.5%,50.5%,22.9%和54.4%,外源喷施褪黑素后,各项指标都明显得到改善。
进一步测定番茄内源褪黑素的含量(图7)和合成关键基因ASMT(图8)的表达发现,低硫条件相对于正常对照而言能够较多的提升番茄内源褪黑素的含量,硫含量(图9)和硫转运蛋白的2个基因SUT1(图10)和SUT2(图11)的表达也显示出外源褪黑素在正常状态下对番茄没有太大的益处,反而是在缺硫条件下能明显促进根部和叶片对硫的吸收,含量分别上升66.1%和74.3%,SUT1和SUT2的表达分别上升1.7/2.2(根/叶)和1.5/1.9倍(根/叶)。
检测硫新陈代谢中的4个关键酶的活性和基因表达可以发现,缺硫处理的番茄在喷施褪黑素后,ATPS、APSR、SiR和OASTL的酶活性分别上调67.3/45.2%、74.6/127.6、26.4/20.5和26.5/23.3%(根/叶,图12),相应基因的表达也明显上调(图13)。
外源施加褪黑素还可以促进番茄体内含硫化合物的合成,如图14~16所示,施加褪黑素后,缺硫处理的番茄Cys、γ-EC和GSH含量都明显上调。以上结果表明,在低硫条件下,外源补充褪黑素制剂能够促进内源褪黑素含量的上升以及对硫的吸收、转运和代谢。
以上所述仅为本发明专利的具体实施案例,但本发明专利的技术特征并不局限于此,任何相关领域的技术人员在本发明的领域内,所作的变化或修饰皆涵盖在本发明的专利范围之中。
Claims (8)
1.一种褪黑素在提高蔬菜硫素吸收中的应用。
2.根据权利要求1所述应用,其特征在于,包括如下步骤:
(1)将褪黑素与附加剂混合后制成褪黑素水剂,所述褪黑素水剂中褪黑素的浓度为25~500μM;
(2)将所述褪黑素水剂喷施于缺硫条件的目标植物叶面或投加于目标植物生长的缺硫营养液或缺硫土壤中。
3.根据权利要求2所述应用,其特征在于,所述附加剂为甲醇、乙醇和水中的至少一种。
4.根据权利要求2所述应用,其特征在于,所述褪黑素水剂中还加入表面活性剂。
5.根据权利要求2所述应用,其特征在于,所述目标植物为番茄。
6.根据权利要求5所述应用,其特征在于,步骤(2)中待番茄长至3叶1心时开始施加所述褪黑素水剂,之后每隔4~6天喷施一次褪黑素水剂,喷施总次数为2~4次。
7.根据权利要求6所述应用,其特征在于,喷施时间为傍晚,喷施量以所有叶片湿润为准。
8.根据权利要求2所述应用,其特征在于,所述褪黑素水剂中褪黑素的浓度为50~150μM。
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