CN115413669B - 激酶抑制剂及其组合剂在提高植物青枯病抗性中的应用 - Google Patents
激酶抑制剂及其组合剂在提高植物青枯病抗性中的应用 Download PDFInfo
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Abstract
本发明属于植物病害防治技术领域,具体涉及激酶抑制剂在提高植物青枯病抗性的应用。以小分子激酶抑制剂库处理拟南芥为基础,初步筛选能够提高拟南芥对青枯菌抗性的小分子激酶抑制剂,然后在青枯病危害较为严重的烟草等植物中验证相关小分子激酶抑制剂的功能,发现小分子激酶抑制剂Paxalisib、CK1‑IN‑1、Ravoxertinib、MLN120B、BX‑912、BQR‑695能够提高烟草对青枯病的抗性,并且不会对烟草的生长产生影响,此外,对抑制剂进行适当组合能够提高抑制剂在青枯病抗性方面的作用效果。
Description
技术领域
本发明属于植物病害防治技术领域,具体涉及激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695在提高植物青枯病抗性的应用。
背景技术
植物在生长发育过程中会遭受各种病原微生物的侵染,进而引发植物的免疫反应,植物能够识别侵染信号,并通过多种方式将信号在细胞间和细胞内进行转导,引起一系列的免疫事件从而实现对侵染的防御。大量研究表明,依赖于蛋白激酶的蛋白质磷酸化与去磷酸化过程在细胞的信号识别与转导中起重要作用,它是生物体中普遍存在的一种调节机制,几乎涉及所有的生理和病理过程,在医学方面已有很多关于蛋白激酶在疾病治疗方面的研究,并且会通过疾病相关激酶的小分子抑制剂进行药物靶向治疗。植物的蛋白磷酸化与去磷酸化研究起步虽然较晚,但随着越来越多蛋白激酶类型的发现,由蛋白激酶所催化的蛋白磷酸化作用在植物细胞信号转导中的地位显得越来越重要,而蛋白激酶的小分子抑制剂在植物免疫过程中的应用还鲜有报道。
激酶抑制剂Paxalisib是一种脑渗透抑制剂,作用于PI3K(PI3Kα,PI3Kβ,PI3Kδ,PI3Kγ)和mTOR。CK1-IN-1是一种新型强效酪蛋白激酶CK1抑制剂,作用于CK1δ和CK1ε、p38ζMAPK。Ravoxertinib是一种新型的、有效的、具有口服生物利用度的细胞外信号调节激酶ERK1/2抑制剂。MLN120B是一种新型、强效、特异性和ATP竞争性IKKβ抑制剂。BX-912是一种新型、有效和特异性的PDK-1抑制剂。BQR-695是一种有效的选择性PI4KIIIβ抑制剂。
青枯菌(Ralstonia solanacearum)是一种毁灭性的植物病原细菌,能侵染50多个科的400多种植物,引起细菌性青枯病,青枯病在辣椒、番茄、马铃薯及茄子等重要茄科类作物上发生和危害尤为严重,给农业生产造成了严重的影响。田间植株一旦感染青枯病,伴随高温高湿的发病有利条件,病株就会迅速发病萎蔫死亡,根据其学术及经济重要性,青枯菌被认为是世界上第二大类的植物病原细菌。到目前为止,对青枯病的防治主要是从化学、生物、农业措施等三大方面进行。化学方面多采用杀线虫剂或熏蒸剂处理土壤来防治烟草青枯病;生物防治主要是从病源的原位土壤中分离出对青枯病具有抑制作用的微生物,制作成微生物有机肥料或者微生物菌剂;农业措施方面包括选用抗病品种、苗床消毒、合理轮作、早播早栽、注意田间卫生和加强田间管理等措施。其中,化学防治是防治青枯病的重要技术措施,与其他防治方法相比,具有快速、有效、方便等特点,但化学防治会导致抗药性、再增猖獗和农药残留等问题;农业措施防治周期较长,操作困难;生物防治效果波动较大,防效不稳定。因此,目前对于青枯病的防治手段还存在多方面的问题,由于根部侵染和维管束定殖的复杂性,使得青枯病的田间防治极具挑战,且青枯菌系变异类型多,侵染来源较为复杂,目前尚无有效的防治措施,开发新的绿色可持续的青枯病防治方法尤为重要。
发明内容
本发明主要解决的技术问题是:克服植物青枯病防治存在的多方面问题,利用植物自身的分子机制进行青枯病的防治,提供一种新的绿色防治方法。
本发明的整体思路为:以小分子激酶抑制剂库为背景,先通过抑制剂对拟南芥进行处理,观察处理后拟南芥对青枯菌抗性的变化,来初步筛选能够提高拟南芥抗青枯菌侵染的小分子激酶抑制剂,然后在青枯病发病较为严重的烟草等植物中验证相关小分子激酶抑制剂的功能,发现六个激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695能够提高植物对青枯菌的抗性,并且不会对植物的生长产生影响,对激酶抑制剂进行适当组合能够提高抑制剂在青枯病抗性方面的作用效果。
本发明的技术方案如下:
通过1400余种激酶活性抑制剂的筛选发现激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695能够有效提高拟南芥对青枯菌的抗性,同时检测这些激酶抑制剂在土壤中对烟草抵抗青枯菌侵染的抗性的调节作用,发现它们能够明显提升烟草对青枯菌的抗性,对不同作用通路的激酶抑制剂进行组合发现抑制剂组合能够提高其在青枯病抗性方面的作用效果。
激酶抑制剂在提高植物青枯病抗性的应用,激酶抑制剂包含Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695中至少一种
激酶抑制剂在制备植物青枯病抑制剂中的应用,激酶抑制剂包含Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695中至少一种。
一种植物青枯病的抑制剂,抑制剂包含激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695中至少一种。
优选地,抑制剂包含激酶抑制剂Paxalisib和CK1-IN-1。
优选地,抑制剂包含激酶抑制剂Paxalisib、Ravoxertinib和BX-912。
优选地,抑制剂中激酶抑制剂有效成分的浓度为10nM-10μM。
优选地,所述激酶抑制剂Paxalisib的有效成分为5-[8,9-二氢-6,6-二甲基-4-吗啉基-6H-(1,4)恶嗪并(4,3-e)嘌呤-2-基]-2-嘧啶胺,CK1-IN-1的有效成分为4-[2-(1-氟-2-萘基)-4-(4-氟苯基)-1H-咪唑-5-基]-吡啶,Ravoxertinib的有效成分为(S)-1-[1-(4-氯-3-氟苯基)-2-羟乙基]-4-{2-[(1-甲基-1H-吡唑-5-基)氨基]嘧啶-4-基}吡啶-2(1H)-one,MLN120B的有效成分为N-[6-氯-7-甲氧基-9H-吡啶多(3,4-b)吲哚-8-基]-2-甲基吡啶-3-甲酰胺,BX-912的有效成分为N-{3-[(5-溴-4-((2-(1H-咪唑-4-基)乙基)氨基)-2-嘧啶基)氨基]苯基}-1-吡咯烷甲酰胺,BQR-695的有效成分为2-{[7-(3,4-二甲氧基苯基)喹喔啉-2-基]氨基}-N-甲基乙酰胺。
优选地,所述激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695有效成分的浓度为10nM-10μM。
优选地,所述植物为易感青枯病的植物。
优选地,所述易感青枯病的植物为烟草、番茄、辣椒。
本发明通过在模式植物拟南芥中筛选能够提高拟南芥对青枯菌的抗性的小分子激酶抑制剂,然后在青枯病危害严重的烟草等植物中验证相关激酶抑制剂的功能,发现激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695能够提高植物对青枯病的抗性;将Paxalisib与CK1-IN-1进行组合发现其对抗性的提升尤为明显,且抑制剂组合也不会直接影响青枯菌的生长。
附图说明
图1为激酶抑制剂Amlexanox的结构式图。
图2为拟南芥野生型Col0被Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695等抑制剂处理与被对照DMSO和无用抑制剂Amlexanox处理相比接种青枯菌后的发病情况。
图3为激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695实验组、对照组的烟草植株生长表型图。
图4为激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695实验组、对照组对烟草抗青枯菌的表型图。
图5为激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695实验组、对照组对青枯菌生长影响的曲线图。
图6为激酶抑制剂组合Paxalisib+CK1-IN-1和Paxalisib+Ravoxertinib+BX-912实验组、对照组对拟南芥抗青枯菌的表型图、烟草植株生长表型图、对烟草抗青枯菌的表型图和对青枯菌生长影响的曲线图。
具体实施方式
表1为小分子激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695的详细信息。
激酶抑制剂Amlexanox的CAS号:68302-57-8,分子式:C16H14N2O4,结构式见图1所示。
实施例1
将一周大小的拟南芥置于含5μM激酶抑制剂的1/2MS液体培养基中培养1d后分为实验组和对照组;实验组添加溶剂DMSO溶解的激酶抑制剂,对照组添加和实验组中等量的DMSO溶剂,之后实验组和对照组接种青枯病菌并使其在培养液中的OD600=0.01,于28℃下长日照培养处理后的拟南芥,每天观察记录其青枯病的病情表型,其中Amlexanox作为在1400余种激酶活性抑制剂筛选过程中其他对青枯病抗性没有明显提升作用的抑制剂的代表,用来作阴性对照,结果如图2所示,相比于DMSO和阴性对照,实验组的抑制剂处理能够明显提高拟南芥对青枯病菌的抗性。
实施例2
激酶抑制剂处理的烟草生长表型鉴定:将每个小盆中加入110g土,每个小盆中分别放入3粒野生型红大烟草种子,分别做5个重复,然后覆盖50mL土,种子萌发后将长势不齐的苗子拔出,留下一株烟草苗。待植物都长大后,实验组采用浇灌10mL含有5μM激酶抑制剂的水溶液,2天后,再次浇灌10mL含有5μM激酶抑制剂的水溶液;对照组在相同时期进行同等处理,只是将激酶抑制剂换成等浓度的DMSO。10天内观察烟草的生长情况,比对对照组和实验组的烟草生长表型。图3说明激酶抑制剂处理不会影响烟草的生长。
实施例3
激酶抑制剂处理的烟草抗青枯病表型鉴定:将每个小盆中加入110g土,每个小盆中分别放入3粒野生型红大烟草种子,分别做5个重复,然后覆盖50mL土,种子萌发后将长势不齐的苗子拔出,留下一株烟草苗。待植物都长大后,实验组浇灌10mL含有5μM激酶抑制剂的水溶液,2天后,再次浇灌10mL含有5μM激酶抑制剂的水溶液,对照组在相同时期进行同等处理,只是将激酶抑制剂换成等浓度的DMSO。一天后,用小刀往近根土壤中切一刀作伤根处理,再加入10mL含有OD600=0.1的青枯病菌CQPS-1。10天内观察烟草的发病情况,比对对照组和实验组的青枯病菌侵染表型。图4说明激酶抑制剂处理能够提高烟草对青枯病菌的抗性。
实施例4
激酶抑制剂处理的青枯菌生长速率统计:将-80℃保存的青枯菌株CQPS-1在固体B培养基上划线,两天后挑取菌落于5mL液体B培养基中,28℃180rpm摇床振荡培养至OD600=1,取菌液100μl分别至5mL含有5μM激酶抑制剂和DMSO的液体B培养基中,于28℃,180rpm摇床振荡,并在12h后,每隔6h测定一次OD600值,绘制实验组和对照组青枯菌的生长速率柱形图。图5说明激酶抑制剂并不会直接影响青枯菌的生长。
实施例5
根据激酶抑制剂的作用通路,将不同通路的激酶抑制剂进行组合实验,第一个抑制剂组合为Paxalisib+CK1-IN-1,第二个组合为Paxalisib+Ravoxertinib+BX-912。对抑制剂组合进行如上述实施例1-4中相同的实验,分别对拟南芥幼苗和烟草进行抑制剂组合处理后接种青枯菌,观察记录抑制剂组合对植物抵抗青枯菌的影响;对烟草进行抑制剂组合的处理,观察抑制剂组合对烟草生长的影响;使用抑制剂组合处理青枯菌,记录抑制剂组合对青枯菌生长的直接影响等实验。从病情指数上看,在接种第10天时,单个激酶抑制剂Paxalisib处理组病情指数为2.2,CK1-IN-1为2.4,Ravoxertinib为2.1,BX-912为2.6,而第一个抑制剂组合Paxalisib+CK1-IN-1病情指数为1.2,是组合中单个抑制剂平均病情指数2.3的52.17%;第二个抑制剂组合Paxalisib+Ravoxertinib+BX-912病情指数为2.0,是组合中单个抑制剂平均病情指数2.3的86.96%,结合图6中表型等其他数据说明抑制剂组合相比于单个抑制剂有着更强的提高植物对青枯菌抗性的能力,第一个抑制剂组合Paxalisib+CK1-IN-1对抗性的提升尤为明显,且抑制剂组合也不会直接影响青枯菌的生长(在图6中Paxalisib简写为Pa、CK1-IN-1简写为CK1、Ravoxertinib简写为Ra、BX-912简写为BX)。
Claims (4)
1.激酶抑制剂在提高植物青枯病抗性中的应用,其特征在于,所述激酶抑制剂包含Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695中至少一种;
所述激酶抑制剂Paxalisib的有效成分为5-[8,9-二氢-6,6-二甲基-4-吗啉基-6H-(1,4)恶嗪并(4,3-e)嘌呤-2-基]-2-嘧啶胺,CK1-IN-1的有效成分为4-[2-(1-氟-2-萘基)-4-(4-氟苯基)-1H-咪唑-5-基]-吡啶,Ravoxertinib的有效成分为(S)-1-[1-(4-氯-3-氟苯基)-2-羟乙基]-4-{2-[(1-甲基-1H-吡唑-5-基)氨基]嘧啶-4-基}吡啶-2(1H)-one,MLN120B的有效成分为N-[6-氯-7-甲氧基-9H-吡啶多(3,4-b)吲哚-8-基]-2-甲基吡啶-3-甲酰胺,BX-912的有效成分为N-{3-[(5-溴-4-((2-(1H-咪唑-4-基)乙基)氨基)-2-嘧啶基)氨基]苯基}-1-吡咯烷甲酰胺,BQR-695的有效成分为2-{[7-(3,4-二甲氧基苯基)喹喔啉-2-基]氨基}-N-甲基乙酰胺。
2.根据权利要求1所述的应用,其特征在于,所述激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695有效成分的浓度为10nM-10μM。
3.激酶抑制剂在制备植物青枯病抑制剂中的应用,其特征在于,所述激酶抑制剂包含Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695中至少一种;
所述激酶抑制剂Paxalisib的有效成分为5-[8,9-二氢-6,6-二甲基-4-吗啉基-6H-(1,4)恶嗪并(4,3-e)嘌呤-2-基]-2-嘧啶胺,CK1-IN-1的有效成分为4-[2-(1-氟-2-萘基)-4-(4-氟苯基)-1H-咪唑-5-基]-吡啶,Ravoxertinib的有效成分为(S)-1-[1-(4-氯-3-氟苯基)-2-羟乙基]-4-{2-[(1-甲基-1H-吡唑-5-基)氨基]嘧啶-4-基}吡啶-2(1H)-one,MLN120B的有效成分为N-[6-氯-7-甲氧基-9H-吡啶多(3,4-b)吲哚-8-基]-2-甲基吡啶-3-甲酰胺,BX-912的有效成分为N-{3-[(5-溴-4-((2-(1H-咪唑-4-基)乙基)氨基)-2-嘧啶基)氨基]苯基}-1-吡咯烷甲酰胺,BQR-695的有效成分为2-{[7-(3,4-二甲氧基苯基)喹喔啉-2-基]氨基}-N-甲基乙酰胺。
4.根据权利要求3所述的应用,其特征在于,所述激酶抑制剂Paxalisib、CK1-IN-1、Ravoxertinib、MLN120B、BX-912、BQR-695有效成分的浓度为10nM-10μM。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1717396A (zh) * | 2002-11-28 | 2006-01-04 | 舍林股份公司 | Chk-、Pdk-和Akt-抑制嘧啶,其制备及作为药物的用途 |
CN103313989A (zh) * | 2010-12-16 | 2013-09-18 | 霍夫曼-拉罗奇有限公司 | 三环pi3k抑制剂化合物和使用方法 |
CN104507926A (zh) * | 2012-03-01 | 2015-04-08 | 阵列生物制药公司 | 丝氨酸/苏氨酸激酶抑制剂 |
WO2015119579A1 (en) * | 2014-02-07 | 2015-08-13 | Agency For Science, Technology And Research | 2,4,5-tri-substituted azole-based casein kinase 1 inhibitors as inducers for cardiomyogenesis |
CN112931508A (zh) * | 2021-01-30 | 2021-06-11 | 湖南大学 | 激酶抑制剂LavendustinA在提高植物青枯病抗性的应用 |
CN113040159A (zh) * | 2021-01-30 | 2021-06-29 | 湖南大学 | 激酶抑制剂Reversine在提高植物青枯病抗性的应用 |
CN113229143A (zh) * | 2021-01-30 | 2021-08-10 | 湖南大学 | 激酶抑制剂Staurosporine在提高植物青枯病抗性的应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
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AU2011201932A1 (en) * | 2010-11-17 | 2012-05-31 | University Of Rochester | Treatment or prevention of fungal infections with PDK1 inhibitors |
-
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- 2022-08-30 CN CN202211060130.5A patent/CN115413669B/zh active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1717396A (zh) * | 2002-11-28 | 2006-01-04 | 舍林股份公司 | Chk-、Pdk-和Akt-抑制嘧啶,其制备及作为药物的用途 |
CN103313989A (zh) * | 2010-12-16 | 2013-09-18 | 霍夫曼-拉罗奇有限公司 | 三环pi3k抑制剂化合物和使用方法 |
CN104507926A (zh) * | 2012-03-01 | 2015-04-08 | 阵列生物制药公司 | 丝氨酸/苏氨酸激酶抑制剂 |
WO2015119579A1 (en) * | 2014-02-07 | 2015-08-13 | Agency For Science, Technology And Research | 2,4,5-tri-substituted azole-based casein kinase 1 inhibitors as inducers for cardiomyogenesis |
CN112931508A (zh) * | 2021-01-30 | 2021-06-11 | 湖南大学 | 激酶抑制剂LavendustinA在提高植物青枯病抗性的应用 |
CN113040159A (zh) * | 2021-01-30 | 2021-06-29 | 湖南大学 | 激酶抑制剂Reversine在提高植物青枯病抗性的应用 |
CN113229143A (zh) * | 2021-01-30 | 2021-08-10 | 湖南大学 | 激酶抑制剂Staurosporine在提高植物青枯病抗性的应用 |
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