拟南芥NO敏感突变体rsn1的特性分析

doi:10.15933/j.cnki.1004-3268.2019.09.006

摘要/Abstract

摘要： 为了深入研究信号分子NO在植物中的调控机制，以拟南芥（Arabidopsis thaliana）为研究材料，从甲基磺酸乙酯(EMS)诱变获得的突变体库中筛选得到1株对一氧化氮（NO）敏感的突变体rsn1（Root sensitive to NO），对比分析突变体根对NO的敏感程度、气孔对脱落酸（ABA）的反应等指标，并且利用图位克隆技术初步定位突变基因。结果表明，尽管在正常生长条件下rsn1突变体的萌发率与野生型没有明显区别，但根对NO胁迫敏感，在50 μmol/L硝普钠(SNP)处理下，野生型拟南芥的相对根长为48.7%，而突变体rsn1的相对根长仅为5.3%；通过分析气孔对ABA的敏感程度发现，rsn1对ABA调控气孔关闭不敏感，在10 μmol/L ABA处理下突变体rsn1的气孔直径是野生型的1.9倍，而在正常条件下突变体rsn1的气孔直径与野生型差异不大，且突变体rsn1的失水率高于野生型拟南芥；遗传分析表明,rsn1是一个单基因隐性突变体；通过图位克隆初步判断突变位点在第5号染色体上。

关键词: 拟南芥, 一氧化氮, rsn1, 脱落酸

Abstract: In order to investigate the molecular regulation mechanism of nitric oxide(NO) in plant，using Arabidopsis as research materials，we screened NO-sensitive mutants and isolated a mutant rsn1 (root sensitive to NO) from the EMS mutant library． The sensitivity of rsn1 mutant roots to NO and stomatal response to abscisic acid(ABA) were compared and analyzed，and the mutant genes were preliminarily located by map-based cloning.The results were as follows: the rsn1 root was more sensitive to NO stress compared with the wild type，though the germination rate of rsn1 mutant was not significantly different from that of wild type under normal growth conditions.Under 50 μmol/L SNP treatment，the relative root length of wild type( as compared with root length in normal growth) was 48.7%，whereas the relative root length of mutant was 5.3%.By analyzing the sensitivity of stomata to ABA，it was found that rsn1 was insensitive to the regulation of stomatal closure by ABA.The stomatal diameter of rsn1 mutant was 1.9 times that of wild type under 10 μmol/L ABA treatment，though the stomatal diameter of rsn1 mutant was similar to that of wild type under normal conditions，and the water loss rate of rsn1 mutant was higher than that of wild type Arabidopsis.Genetic analysis suggested that rsn1 was a single recessive mutant，and the mutation site was initially designated on chromosome 5 by map-based cloning．

Key words: Arabidopsis thaliana, Nitric oxide; rsn1, ABA

中图分类号:

S355.3

夏金婵, 从人愿, 张小莉. 拟南芥NO敏感突变体rsn1的特性分析[J]. 河南农业科学, 2019, 48(9): 40-45.

XIA Jinchan, CONG Ｒenyuan, ZHANG Xiaoli. Characterization of Nitric Oxide Sensitive Mutant rsn1 in Arabidopsis[J]. Journal of Henan Agricultural Sciences, 2019, 48(9): 40-45.

参考文献

［1］HUANG T F,JANDER G.Abscisic acidregulated protein degradation causes osmotic stressinduced accumulation of branchedchain amino acids in Arabidopsis thaliana［J］.Planta,2017,246(4):737-747.

［2］MA Y,SZOSTKIEWICZ I,KORTE A,et al.Regulators of PP2C phosphatase activity function as abscisic acid sensors［J］.Science,2009,324(5930):1064-1068. 

［3］PARK S Y,FUNG P,NISHIMURA N,et al.Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins［J］.Science,2009,324(5930):1068-1071.

［4］SCHWEIGHOFER A,HIRT H,MESKIENE I.Plant PP2C phosphatases:Emerging functions in stress signaling［J］.Trends in Plant Science,2004,9(5):236-243.

［5］FUJITA Y,NAKASHIMA K,YOSHIDA T,et al.Three SnRK2 protein kinases are the main positive regulators of abscisic acid signaling in response to water stress in Arabidopsis［J］.Plant and Cell Physiololgy,2009,50(12):2123-2132.

［6］WANG J L,ZHANG Q,YU Q,et al.CARK6 is involved in abscisic acid to regulate stress responses in Arabidopsis thaliana［J］.Biochem Biophys Res Commun,2019,513(2):460-464.

［7］CORPAS F J,BARROSO J B,CARRERAS A,et al.Constitutive argininedependent nitric oxide synthase activity in different organs of pea seedlings during plant development［J］.Planta,2006,224(2):246-254.

［8］ZHAO Z G,CHEN G C,ZHANG C L.Interaction between reactive oxygen species and nitric oxide in droughtinduced abscisic acid synthesis in root tips of wheat seedlings［J］.Aust J Plant Physiol,2001,28(10):1055-1061.

［9］LESHEM Y Y,HARAMATY E.Plant aging:The emission of NO and ethylene and the effect of NOreleasing compounds on growth of pea(Pisum sativum) foliage［J］.J Plant Physiol,1996,148:258-263.

［10］ALLEN G J,CHU S P,HARRINGTON C L,et al.A defined range of guard cell calcium oscillation parametersencodes stomatal movements［J］.Nature,2001,411(6841):1053-1057.

［11］LEE Y,CHOI Y B,SUH S,et al.Abscisic acidinduced phosphoinositide turnover in guard cellprotoplasts of Vicia faba［J］.Plant Physiol,1996,110(3):987-996.

［12］MCAINSH M R,BROWNLEE C,HETHERINGTON A M.Abscisic acidinduced elevation of guard cell cytosolic Ca²⁺precedes stomatal closure［J］.Nature,1990,343(6254):186-188.

［13］WANG P C,DU Y Y,HOU Y J,et al.Nitric oxide negatively regulates abscisic acid signaling in guard cells by Snitrosylation of OST1［J］.Proc Natl Acad Sci USA,2015,112(2):613-618.

［14］DESIKAN R,GRIFFITHS R,HANCOCK J,et al.A new role for an old enzyme:Nitratereductasemediated nitric oxide generation is required for abscisic acidinduced stomatal closure in Arabidopsis thaliana［J］.Proc Natl Acad Sci USA,2002,99(25):16314-16318.

［15］LOZANOJUSTE J,LEN J.Enhanced abscisic acidmediated responses in nia1nia2noa12 triple mutant impaired in NIA/NR and AtNOA1dependent nitric oxide biosynthesis in Arabidopsis［J］.Plant Physiol,2010,152(2):891903.

［16］SONG C P,AGARWAL M,OHTA M,et al.Role of an Arabidopsis AP2/EREBPtype transcriptional repressor in abscisic acid and drought stress responses［J］.Plant Cell,2005,17(8):2384-2396.

［17］LUKOWITZ W,GILLMOR C S,SCHEIBLE W R.Positional cloning in Arabidopsis.Why it feels good to have a genome initiative working for you［J］.Plant Physiology,2000,123(3):795-805.

［18］HE Y K,TANG R H,HAO Y,et al.Nitric oxide represses the Arabidopsisfloral transition［J］.Science,2004,305(5692):1968-1971.

［19］SIDDIQUI M H,ALWHAIBI M H,BASALAH M O.Role of nitric oxide in tolerance of plants to abiotic stress［J］.Protoplasma,2011,248(3):447-455.

［20］FANCY N N,BAHLMANN A K,LOAKE G J.Nitric oxide function in plant abiotic stress［J］.Plant Cell and Environment,2017,40(4):462-472.

［21］FAN Q J,LIU J H.Nitric oxide is involved in dehydration/drought tolerance in Poncirus trifoliata seedlings through regulation of antioxidant systems and stomatal response［J］.Plant Cell Reports,2012,31(1):145-154.

［22］KALE L,NAKURTE I,JALAKAS P,et al.Arabidopsis mutant dnd2 exhibits increased auxin and abscisic acid content and reduced stomatal conductance［J］.Plant Physiol Biochem,2019,140:18-26.

［23］NEILL S J,DESIKAN R,CLARKE A,et al.Nitric oxide is a novel component of abscisic acid signaling in stomatal guard cells［J］.Plant Physiol,2002,128(1):13-16.

［24］GARCIAMATA C,GAY R,SOKOLOVSKI S,et al.Nitric oxide regulates K⁺ and Cl^- channels in guard cells through a subset of abscisic acidevoked signaling pathways［J］.Proc Natl Acad Sci USA,2003,100(19):11116-11121.

［25］VAHISALU T,KOLLIST H,WANG Y F,et al.SLAC1 is required for plant guard cell Stype anion channel function in stomatal signalling［J］.Nature,2008,452(7186):487-491.

［26］JOUDOI T,SHICHIRI Y,KAMIZONO N,et al.Nitrated cyclic GMP modulates guard cell signaling in Arabidopsis［J］.Plant Cell,2013,25(2):558-571.

［27］杨凤博,王鲜萍,李坤.拟南芥蛋白激酶SNRK2.6的原核表达、纯化及活性分析［J］.河南农业科学,2014,43(10):26-29.

［28］雷凯健,王棚涛,刘浩.拟南芥响应过氧化氢突变体的筛选及遗传分析［J］.河南农业科学,2015,44(10):53-56.

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