3种外源诱导剂预处理对甜瓜抗病特征及其Fom-2和CHT基因表达的影响
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摘要
该研究选用水杨酸(SA)、茉莉酸甲酯(MeJA)、Ca~(2+)、无菌水(对照)作为外源预处理诱导剂,以抗、感枯萎病甜瓜品种为材料,分别于诱导预处理2d后接种甜瓜枯萎菌,并于接种5、7、9d时观察发病情况,进行病情调查;在接种后1、3、5、7、9d取甜瓜叶片,分析抗病甜瓜(MR-1)和感病甜瓜(M1-15)叶片中甜瓜抗枯萎病基因(Fom-2)、几丁质酶基因(CHT)的表达变化,以探寻提高防治甜瓜枯萎病菌侵染的技术途径。结果显示:(1)外源MeJA和SA预处理接种后2品种的病情指数显著低于对照,但Ca~(2+)处理后的病情指数与对照无显著差异。(2)经外源诱导预处理接种后,MR-1和M1-15品种叶片的Fom-2和CHT基因均出现差异表达,但Ca~(2+)诱导其上调表达的效果微弱。(3)经SA、MeJA诱导预处理接种后,2品种叶片的Fom-2和CHT基因表达总体均显著高于对照;Fom-2基因的表达抗病甜瓜MR-1分别在接种后5d、7d时达到峰值,而感病甜瓜M1-15则均在接种9d时达到峰值;CHT基因的表达抗病甜瓜MR-1则均在接种后7d时达到峰值,而感病甜瓜M1-15分别在接种后7d、9d时达到峰值。(4)Ca~(2+)处理对抗、感甜瓜叶片的Fom-2和CHT基因的表达均无显著影响。(5)相关分析表明,经SA、MeJA诱导预处理接种后,甜瓜枯萎病病情指数与Fom-2和CHT基因表达量有显著的相关性;而Ca~(2+)处理效果不显著。研究表明:SA、MeJA通过诱导Fom-2、CHT基因上调表达,进而使甜瓜的抗病性提高,而Ca~(2+)处理对两基因表达和甜瓜抗病性均无显著影响。
In this study,salicylic acid(SA),methyl jasmonate(MeJA),Ca~(2+),and sterile water as a control were used as exogenous inducers to pretreat the melons for resistance and susceptibility to fusarium wilt.Two days after the induction of pretreatment,they were inoculated with F.melilocarpa,and the incidence was observed on the 5,7 and 9 dafter inoculation.The disease status was investigated,and melon leaves were taken at 1,3,5,7 and 9 dafter inoculation.Expression changes of Fom-2 and CHTin resistant melon(MR-1)and susceptible melon(M1-15)leaves were evaluated in order to check the wilt in con-trol wilt with the technical way of dyeing.The results showed that:(1)the disease indexes of the two varieties after inoculation with exogenous MeJA and SA were significantly lower than that of the control and there was no significant difference between the condition index after Ca~(2+)treatment and the control.(2)After inoculated with exogenously induced pretreatment,the expression of Fom-2 and CHTgenes in leaves of MR-1 and M1-15 cultivars was different but the effect of Ca~(2+)on upregulation was not strong.(3)After inoculated with SA and MeJA,the expression of Fom-2 and CHTgenes in the leaves of the two cultivars were significantly higher than those of the control,but the expression level of Fom-2 for resistant melon MR-1,the peaks were reached at 5 dand 7 dand all susceptible melon M1-15 were peaked at 9 dafter inoculation.The expression of CHT gene for resistant melon MR-1 peaked at 7 dafter inoculation.M1-15 showed peak at 7 and 9 days after inoculation,respectively.(4)There was no significant effect of Ca~(2+)treatment on the expression of Fom-2 and CHTgenes in muskmelon leaves.(5)The correlation analysis showed that the disease index of Fusarium wilt was correlated with the expression levels of Fom-2 and CHTgenes after SA and MeJA induced pretreatment inoculation.The effect of Ca~(2+)treatment is not significant.
In this study,salicylic acid(SA),methyl jasmonate(MeJA),Ca~(2+),and sterile water as a control were used as exogenous inducers to pretreat the melons for resistance and susceptibility to fusarium wilt.Two days after the induction of pretreatment,they were inoculated with F.melilocarpa,and the incidence was observed on the 5,7 and 9 dafter inoculation.The disease status was investigated,and melon leaves were taken at 1,3,5,7 and 9 dafter inoculation.Expression changes of Fom-2 and CHTin resistant melon(MR-1)and susceptible melon(M1-15)leaves were evaluated in order to check the wilt in con-trol wilt with the technical way of dyeing.The results showed that:(1)the disease indexes of the two varieties after inoculation with exogenous MeJA and SA were significantly lower than that of the control and there was no significant difference between the condition index after Ca~(2+)treatment and the control.(2)After inoculated with exogenously induced pretreatment,the expression of Fom-2 and CHTgenes in leaves of MR-1 and M1-15 cultivars was different but the effect of Ca~(2+)on upregulation was not strong.(3)After inoculated with SA and MeJA,the expression of Fom-2 and CHTgenes in the leaves of the two cultivars were significantly higher than those of the control,but the expression level of Fom-2 for resistant melon MR-1,the peaks were reached at 5 dand 7 dand all susceptible melon M1-15 were peaked at 9 dafter inoculation.The expression of CHT gene for resistant melon MR-1 peaked at 7 dafter inoculation.M1-15 showed peak at 7 and 9 days after inoculation,respectively.(4)There was no significant effect of Ca~(2+)treatment on the expression of Fom-2 and CHTgenes in muskmelon leaves.(5)The correlation analysis showed that the disease index of Fusarium wilt was correlated with the expression levels of Fom-2 and CHTgenes after SA and MeJA induced pretreatment inoculation.The effect of Ca~(2+)treatment is not significant.
引文
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[2]赵惠新,李冠,李金玉,等.甜瓜抗病基因的研究进展及其应用前景[J].分子植物育种,2005,(2):249-254.ZHAO H X,LI G,LI J Y,et al.Development of research about resistance genes cloning in melon(Cucumis melo L.)and the applications foreground[J].Molecular Plant Breeding,2005,(2):249-254.
[3]陈爱葵,庄文宋,陈冬梅.植物几丁质酶及其基因工程研究进展[J].广东教育学院学报,2010,30(3):47-53.CHEN A K,ZHUANG W S,CHEN D M.Development of Research about chitinase and its genetic engineering in plant[J].Journal of Guangdong Education Institute,2010,30(3):47-53.
[4]SCHLUMBAUM A,MAUCH F,VOGELI U,et al.Plant chitinas-es are potentinhibitors of fungal growth[J].Nature,1986,324(27):365-367.
[5]MARCHENT R,DAVERY MR,LUCAS JA,et al.Expression of achitinase transgene in rose(Rosa hybrida L.)reduces de-velopment of blackspot disease(Diplocarpon rosae wolf)[J].Molecular Breeding,1998,4(3):187-194.
[6]TURNER J G,ELLIS C,DEVOTO A.The jasmonate signal pathway[J].Plant Cell,2002,14(Suppl):S153.
[7]RASKIN I.Role of salicylic acid in plants[J].Annu.Rev.Plant Physiol.Plant Mol.Biol.,2003,43:439-463.
[8]LIECHTI R,FARMER E E.The jasmonate pathway[J].Science,2002,296(5 573):1 649.
[9]石延霞,于洋,傅俊范,等.病原菌诱导后黄瓜叶片中脂氧合酶活性与茉莉酸积累的关系[J].植物保护学报,2008,35(6):486-490.SHI Y X,YU Y,FU J F,et al.Relationship between LOX activity and JA accumulations in cucumber leaves induced by pathogen[J].Journal of Plant Protection,2008,35(6):486-490.
[10]冯远娇,王建武,苏贻娟,等.茉莉酸在玉米地上部与地下部系统性诱导防御反应中的作用[J].中国农业科学,2009,42(8):2 726-2 735.FENG Y J,WANG J W,SU Y J,et al.The role of jasmonic acid in the systemic induced defense response of aboveground and underground in corn(Zea mays)[J].Scientia Agriculture Sinica,2009,42(8):2 726-2 735.
[11]吴国昭,谢丽君,宋圆圆,等.外源信号物质诱导广东高州普通野生稻抗稻瘟病的生理生化机理[J].西北农业学报,2009,18(3):254-258.WU G Z,XIE L J,SONG Y Y,et al.The physiological and biochemical mechanism of disease resistance of Oryzarufipogon griff in digenous to Gaozhou,Guangdong against Pyriculariagriseainduced by exogenousily applied plant signal compounds(in Chinese)[J].Acta Agriculturae Borealioccidentalis Sinica,2009,18(3):254-258.
[12]张建成,范永山,董金皋.玉米大斑病菌毒素对玉米叶肉细胞脂氧合酶活性的影响[J].植物病理学报,2003,33(5):421-424.ZHANG J C,FAN Y S,DONG J G,et al.Effects of HTtoxin produced by Exserohilum turcicum on lip oxygenase activity in corn leaf cells[J].Acta Phytopathologica Sinica,2003,33(5):421-424.
[13]SCHAFFRATH U,SCHEINPFLUG H,REISENER H J.An elicitor fromPyriculariaoryzaeinduces resistance responses in rice:isolation,characterization and physiological properties[J].Acta Phytopathologica Sinica,1995,46(4):293-307.
[14]陈利锋,聂理.抗感赤霉病小麦品系超氧化物歧化酶和过氧化氢酶的活性比较[J].植物病理学报,1997,27(3):209-212.CHEN L F,NIE L.Comparison of Scab Resistant and susceptible wheat varieties of superoxide dismutase and catalase activity[J].Acta Phytopathologica Sinica,1997,27(3):209-212.
[15]张智慧,聂燕芳,何磊,等.外源茉莉酸甲酯诱导水稻抗瘟性相关防御酶和内源水杨酸的变化[J].植物病理学报,2010,40(4):395-403.ZHANG Z H,NIE Y F,HE L,et al.Resisance-related defense enzymes and endogenous salicylic acid induced by exogenous methyl jasmonae in rice against blast disease[J].Acta Phytopathologica Sinica,2010,40(4):395-403.
[16]宾金华,姜胜,黄胜琴,等.茉莉酸甲酯诱导烟草幼苗抗炭疽病与PAL活性及细胞壁物质的关系[J].植物生理学报,2000,26(1):2-7.BIN J H,JIANG S,HUANG S Q,et al.Relationship between anthracnose resistance and PAL activity and cell wall substance in tobacco seedlings induced by methyl jasmonate[J].Physiological&Molecular Plant Pathology,2000,26(1):2-7.
[17]向妙莲,付永琦,何永明,等.茉莉酸甲酯浸种对水稻幼苗白叶枯病抗性及抗氧化酶活性的影响[J].中国水稻科学,2014,28(4):419-426.XIANG M L,FU Y Q,HE Y M,et al.Effects of seed soaking with Methyl Jasmonate on resistance to bacterial leaf blight and antioxidant enzyme activities of rice seedlings[J].Chinese Journal of Rice Science,2014,28(4):419-426.
[18]范存斐,毕阳,王云飞,等.水杨酸对厚皮甜瓜采后病害及苯丙烷代谢的影响[J].中国农业科学,2012,45(3):584-589.FAN C F,BI Y,WANG Y F,et al.Effect of salicylic acid dipping on postharvest diseases and phenylpropanoid pathway in muskmelon fruits[J].Scientia Agricultura Sinica,2012,45(3):584-589.
[19]曾凯芳,姜微波.水杨酸处理对采后绿熟芒果炭疽病抗病性的诱导[J].中国农业大学学报,2005,10(2):36-40.ZENG K F,JIANG W B.Induced resistance to anthracnose(Colletotichum gloeosporioides)in mango fruit by postharvest treatment with salicylic acid[J].Journal of China Agricultural University,2005,10(2):36-40.
[20]徐丙寅.甜瓜抗枯萎病基因Fom-2的表达特性分析与功能验证[D].哈尔滨:东北农业大学,2013.
[21]武兴丽.瓜类枯萎病的发生与防治[J].北方园艺,2003,(5):51.WU X L.Occurrence and control of cucurbit wilt disease[J].Northern Horticulture.2003,(5):51.
[22]包海清,许勇,杜永臣,等.海南三亚地区地区葫芦科作物白粉病菌生理小种分化的鉴定[J].长江蔬菜,2008,(1):49-51.BAO H Q,XU Y,DU Y C,et al.Identification of physiological races of powdery mildew on cucurbits in Sanya,Hainan[J].Journal of Changjiang Vegetables,2008,(1):49-51.
[23]刘璐,孙蕾,张志鹏,等.外源物质诱导对甜瓜枯萎病抗性和防御酶活性的影响[J].北方园艺,2016,(14):122-126.LIU L,SUN L,ZHANG Z P,et al.Effects of exogenous substance induction on Fusarium wilt resistance and defense enzyme activity in melon[J].Northern Horticulture,2016,(14):122-126.
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