CN107396935B - 硫化氢供体硫氢化钠在抑制植物器官脱落中的应用 - Google Patents
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Abstract
本发明公开了硫化氢(H2S)供体硫氢化钠在抑制植物器官脱落中的应用。本发明利用H2S气体(由硫氢化钠液体挥发提供)熏蒸拟南芥和番茄植株，可显著延迟拟南芥花器官的脱落与果荚开裂，显著抑制番茄花柄的脱落；H2S可减弱脱落促进剂乙烯对脱落的促进作用；在H2S处理过程中检测器官脱落过程标志蛋白(多聚半乳糖醛酸酶、纤维素酶和细胞壁松弛蛋白)编码基因的表达水平，发现与未处理的对照组不同，H2S处理的花柄中相关基因的表达量上升迟缓，说明H2S在分子水平抑制脱落过程的发生。本发明试剂单一易获取，操作简单，为缓解植物器官脱落提供了一种实用有效的方法。
Description
技术领域
本发明涉及硫氢化钠的新用途，准确地说是硫化氢供体硫氢化钠作为抑制剂延迟植物器官脱落的新应用。
背景技术
器官脱落，是指植物细胞、组织或器官脱离母体的生理过程。它是植物应对外界环境(如病原物)或放弃那些不再有用途的多余器官的有效机制。落花、落叶、落果以及果荚开裂等都属于器官脱落的范畴。同时器官脱落又是一个重要的农艺性状，在生产中，当作物遭受干旱、冷、以及病虫害等胁迫时，为避免这些不利因素所带来的产量上的影响，人们需要抑制器官脱落的发生。因此，控制器官脱落发生的时间一直是植物学家、农学家和园艺学者们的研究热点。植物器官脱落过程一般可分为四步：(1)离区的形成；(2)获取响应脱落信号的能力，(3)脱落的激活，以及(4)保护层的形成。植物器官在脱落之前，需要多聚半乳糖醛酸酶(PG)、纤维素酶(Cel)以及细胞壁松弛蛋白(EXP)等降解细胞间纤维素和果胶的酶类上调表达，使离区细胞之间彼此分离，进而达到促进器官脱落的目的，因而这些酶类编码基因的上调表达往往是植物器官要发生脱落的标志。各种植物激素也参与到植物器官脱落中，其中生长素和乙烯的作用最为显著：生长素抑制脱落，而乙烯则是脱落促进剂。在番茄中，盛花期的花柄由花器官提供生长素，抑制花柄脱落；但当花器官被摘除后，花柄的脱落过程被启动。目前，研发改良品种是延迟植物器官脱落的最常用方法，但新品种的改良耗时较长，研究手段也很复杂，对结果的预期不能及时把控。
硫化氢(H2S)是近年来继NO和CO之后发现的第三种气体信号分子，其功能已在动物和植物中有诸多阐述。在动物中，生理浓度的H2S具有保护心脏、舒张血管，促进微血管内皮细胞增殖、诱导血管平滑肌细胞凋亡，改善糖尿病、败血症、肝硬化、心脏衰竭、哮喘等多种调节功能。而在植物中，H2S能够促进种子的萌发和不定根的生成，提高光合效率，促进植物的生长发育，可以帮助植物抵抗干旱胁迫，增强植物对重金属和热胁迫的耐受性等。目前关于硫化氢在植物体内生理功能的研究，主要通过用H2S的药物供体处理植物材料，观察和测定植物生理指标变化。最常用的H2S供体有硫氢化钠(NaHS)和GYY4137，两者溶于水后均可产生H2S气体；可以释放H2S的供体还有CaS、Na2S等。它们的反应机理是：NaHS+H2O＝Na++HS-+H2O＝H2S+NaOH。此外，人们还利用各种硫化氢生成抑制剂或清除剂来进行材料处理，观察硫化氢的合成被抑制或是硫化氢被清除以后受体材料所产生的相应变化，进而分析硫化氢的功能。目前常用的硫化氢合成抑制剂是羟胺(HA)，常用的清除剂是HT(次牛磺酸)。最近我们研究发现，气体H2S还可以抑制植物器官脱落。
发明内容
本发明旨在提供硫化氢供体硫氢化钠在抑制植物器官脱落中的应用。
本发明提供的硫化氢供体硫氢化钠在抑制植物器官脱落中的应用。
所述硫氢化钠为30-100umol/L的NaHS水溶液。
所述植物器官为拟南芥花器官、果荚或番茄花柄。
所述硫氢化钠可以用硫化钠、硫化钙或GYY4137替代。
本发明将硫氢化钠作为气体信号分子硫化氢的提供者，其外源的施用浓度很低，仅为30-100umol/L的NaHS水溶液；NaHS水溶液释放出的微量H2S气体处理活体植物，可明显延迟植物器官的脱落。
附图说明
图1 H2S延迟拟南芥花器官脱落与果荚开裂
图2 H2S抑制自然衰老多引起的番茄花柄脱落
图3 H2S及其合成抑制剂与清除剂对番茄花柄脱落的影响
图4 H2S抑制细胞壁降解相关基因的上调表达
具体实施方式
以下给出本发明的非限定实施例。
实施例1
取两组生长状态一致的开花期拟南芥植株，其中一组每天用50μm的H2S(在7L的罩子中放置一个盛有350μl 1M NaHS液体的盖子)熏蒸6小时，另外一组不做处理(对照组)。观察两组拟南芥花的脱落情况。结果显示，H2S处理组拟南芥花器官的脱落明显晚于对照组。说明在拟南芥中，H2S可以抑制花器官的脱落。
取两组生长状态一致的已结荚的拟南芥植株，其中一组每天用50μm的H2S(在7L的罩子中放置一个盛有350μl 1M NaHS液体的盖子)熏蒸6小时，另外一组不做处理(对照组)。观察两组拟南芥果荚的开裂情况。结果显示，H2S处理组拟南芥果荚的开裂明显晚于对照组。说明在拟南芥中，H2S可以抑制果荚的开裂(见附图1)。
实施例2
取两组生长状态一致的生殖生长期番茄植株，其中一组每天用50μm的H2S(在7L的罩子中放置一个盛有350μl 1M NaHS液体的盖子)熏蒸6小时，另外一组不做处理(对照组)。观察番茄花柄脱落情况，结果显示，H2S处理组番茄花柄脱落比例明显低于对照组(见附图2)。
实施例3
将番茄花柄摘除花器官以后，分为四组，插到0.7％的固体琼脂上。
这四组花柄分别为对照、硫化氢处理、羟胺(HA)处理和次牛磺酸(HT)处理。
称取硫氢化钠加水配制50μmol/L浓度水溶液，至于离心管盖子中，放置于盛有固体琼脂的培养皿中；同时将1ml HA(100μm)和1ml HT(100μm)分别加到各自培养皿的琼脂上，将培养皿盖子封好，观察花柄脱落情况。
结果显示：硫化氢可以抑制番茄花柄脱落，未经处理的番茄花柄在摘下来以后12小时开始脱落，而硫化氢处理的花柄24小时还未开始脱落；而硫化氢生成抑制剂HA和清除剂HT则促进脱落，HT处理8小时，花柄已开始脱落，而HA处理10小时，花柄也已开始脱落。(见附图3)
实施例4
取开花期摘除花器官的花柄(为保证花柄的状态一致)，将其置于1/2MS培养基上，分对照组和H2S处理组，其中处理组用H2S(以NaHS水溶液为供体)处理12h后涂抹乙烯利，而对照组前12h不做任何处理，12h后和处理组一同用乙烯利处理。实验结果显示，乙烯利处理9h后，这些处理中的花柄脱落率为零，而对照组的花柄脱落率为100％，说明H2S可抑制乙烯所引起的番茄花柄脱落。我们进一步分析了实验花柄中与细胞壁降解有关的基因(多聚半乳糖醛酸酶、纤维素酶和细胞壁松弛蛋白编码基因)的表达情况：与对照组相比，H2S处理组中这些基因的表达量在乙烯利处理前后上调都比较迟缓，表明H2S处理之后，番茄花柄获取脱落信号和激活脱落的能力减弱了，说明H2S参与花柄脱落的中间两个步骤，并且可能同花柄中生长素和乙烯的作用有关系。(见附图4)
实施例5
取6组生长状态一致的生殖生长期植株(未摘除花器官，生长素供给正常)，按照表1中的设计处理。实验结果显示，当与乙烯利共同处理时，H2S可以抑制乙烯对花柄脱落的促进作用(第2组)，而HT会加快花柄脱落(第5组)，但有意思的是H2S预处理在本实验中对花柄脱落没有作用(第3组)，说明H2S在生长素供应正常的情况下，可能对生长素含量的影响不大，而且H2S可能与乙烯下游信号传导也有关系。表1如下：
表1 H2S同乙烯互作关系分析实验设计与结果
| 处理方法 | 花柄脱落开始时间 | |
| 第1组 | 乙烯利处理 | 8h |
| 第2组 | 乙烯利与H<sub>2</sub>S同时处理 | 12h |
| 第3组 | 先用H<sub>2</sub>S预处理，再喷洒乙烯利溶液 | 8h |
| 第4组 | 乙烯利处理1h后用H<sub>2</sub>S处理 | 12h |
| 第5组 | 乙烯利与H<sub>2</sub>S清除剂同时处理 | 6h |
| 第6组 | 对照组(喷洒清水) | 无脱落发生 |
注：脱落时间统计从喷洒乙烯利时开始计算。
Claims (2)
1.硫化氢供体硫氢化钠在抑制植物器官脱落中的应用；所述硫氢化钠为30-100umol/L的NaHS水溶液；所述植物器官为拟南芥花器官、果荚或番茄花柄。
2.如权利要求1所述的硫化氢供体硫氢化钠在抑制植物器官脱落中的应用，所述硫氢化钠用硫化钠、硫化钙或GYY4137替代。
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| US4086079A (en) * | 1976-04-14 | 1978-04-25 | The Procter & Gamble Company | Sulfides and derivatives thereof as abscission agents |
| CN102550652A (zh) * | 2012-01-10 | 2012-07-11 | 合肥工业大学 | 一种硫化氢供体硫氢化钠促进果蔬储藏保鲜的新用途 |
| CN104304316A (zh) * | 2014-10-20 | 2015-01-28 | 山西大学 | 硫化氢供体硫氢化钠促进花卉开花并延长花期的新用途 |
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| US4086079A (en) * | 1976-04-14 | 1978-04-25 | The Procter & Gamble Company | Sulfides and derivatives thereof as abscission agents |
| CN102550652A (zh) * | 2012-01-10 | 2012-07-11 | 合肥工业大学 | 一种硫化氢供体硫氢化钠促进果蔬储藏保鲜的新用途 |
| CN104304316A (zh) * | 2014-10-20 | 2015-01-28 | 山西大学 | 硫化氢供体硫氢化钠促进花卉开花并延长花期的新用途 |
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