Harpin处理对苹果梨黑斑病的抑制及抗性酶的诱导

研究了采后Harpin真空渗透处理对苹果梨(Pyrus bretschneideri‘Pingguo’)损伤接种互隔交链孢(Alternar-ia alternata)的影响及部分机理.结果表明:用50、100和150 mg/L的Harpin处理均可明显抑制损伤接种果实的发病率,其中以50 mg/L Harpin的效果最好.此外,该浓度处理还明显降低损伤接种果实的病斑面积.苹果梨经50 mg/LHarpin处理再用(A.alternata)挑战后果实体内的过氧化物酶(POD)、几丁质酶(CHT)和苯丙氨酸解氨酶(PAL)活性明显升高,酚类物质的含量也显著积累.由此表明,Harpin对苹果梨黑斑病有效抑制与诱导果实体内这3种抗性酶的活性以及提高酚类物质含量密切相关.     

表达 HarpinEa基因的转基因马铃薯的晚疫病抗性分析

 通过根癌农杆菌介导,用Harpin_Ea基因转化马铃薯四倍体栽培种大西洋（Atlantic）,获得107个马铃薯转化株系。经过卡那霉素的抗性鉴定和PCR分子检测,有59株PCR检测成阳性,占所有转化植株的55.14%.实验对部分转基因植株进行室内抗病性评价,从24个PCR阳性株系中筛选到9个株系的抗病性与对照相比有显著差异(P<0.05),其中7株的抗病性与对照相比差异极显著(P<0.001)。结果初步表明Harpin_Ea基因已整合到马铃薯的基因组中并得到表达,提高了植株的晚疫病抗性。     

超敏蛋白诱导水稻对立枯病的抗性

[目的]明确超敏蛋白对水稻立枯病的诱抗作用。[方法]以龙粳37水稻品种为试验材料,研究不同浓度超敏蛋白浸种处理对水稻生长发育的影响,测定超敏蛋白处理后水稻苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)含量变化。[结果]不同浓度超敏蛋白对水稻生长发育均有促进作用,且与对照相比不同浓度超敏蛋白处理水稻均不同程度地诱导了水稻防御酶PAL、POD和PPO活性,以稀释20倍的超敏蛋白诱导能力最强。[结论]研究结果为超敏蛋白抗病机理研究提供了理论依据。     

Harpin防治大豆灰斑病效果的研究

Harpin对大豆灰斑病具有较好防效,防治效果平均为46.7%,与生产上常用杀菌剂50%多菌灵可湿性粉剂防效相当,其最适用量为2 L·hm-2,该生物药剂对作物安全.     

水稻条斑病细菌类Harpin蛋白的纯化与特性研究

 用硫酸铵沉淀、制备等电聚焦电泳、阴离子交换层析等方法从水稻条斑病细菌(Xanthomonas oryzae pv. oryzicola,Xooc) RS105菌株突变体M51菌体破碎液中纯化出可激发烟草产生过敏性反应(hypersensitive response,HR)的蛋白类物质,分子量约为25.5 kDa。该物质与梨火疫病菌(Erwinia amylovora))的HarpinEa和水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,Xoo)的Harpin_Xoo具有相似的生物活性和理化特性:可激发烟草产生典型的HR反应;对热稳定,对蛋白酶K敏感;RNA转录抑制剂放线菌素D、蛋白质合成抑制剂环己酰亚铵和钙离子通道阻断剂氯化镧能够抑制该物质激发烟草产生HR;该物质具有诱导烟草抗TMV的功能。据此,将该物质命名为类Harpin蛋白(Harpin-like protein,HLP_Xooc)。     

Harpin蛋白的研究进展

由植物革兰氏阴性病原菌编码产生的harpin蛋白是激发植物过敏反应的主要因子，具有显著的诱导植物抗性和促进生长发育等多种生物学效应，同时也是研究植物抗病信号传导的理想模型。基于对harpin蛋白的深入研究，国外已开发出新型、高效、安全的生物农药。本文综述了近年来harpin蛋白的研究进展。     

Evaluation of postharvest treatments with chemical resistance inducers to control green and blue molds on orange fruit

Preventive and curative activities of postharvest treatments with selected chemical resistance inducers to control postharvest green (GM) and blue (BM) molds on oranges (cvs. ‘Valencia’ or ‘Lanelate’) artificially inoculated with Penicillium digitatum and Penicillium italicum, respectively, were evaluated. In vivo primary screenings to select the most effective chemicals and concentrations were performed with benzothiadiazole (BTH), β-aminobutyric acid (BABA), 2,6-dichloroisonicotinic acid (INA), sodium silicate (SSi), salicylic acid (SA), acetylsalicylic acid (ASA) and harpin. INA at 0.03 mM, SA at 0.25 mM, BABA at 0.3 mM and BTH at 0.9 mM were selected and tested afterwards as dips at 20 °C for 60 or 150 s with oranges artificially inoculated before or after the treatment and incubated for 7 d at 20 °C. Although it was an effective treatment, SSi at 1000 mM was discarded because of potential phytotoxicity to the fruit rind. Preventive or curative postharvest dips at room temperature had no effect or only reduced the development of GM and BM very slightly. Therefore, these treatments cannot be recommended for inclusion in postharvest decay management programs for citrus packinghouses.     

Harpin蛋白基因的克隆及序列分析

 以梨火疫病细菌基因组DNA为模板,通过合成5'-端及3'-端一对特异引物,利用多聚酶链式反应(PCR)扩增获得harpin蛋白基因。将其克隆到E.coli质粒上进行序列分析。结果表明:基因由1155个核苷酸组成,编码385个氨基酸残基组成的多肽。与发表序列相比较,核苷酸序列及推导出的氨基酸序列的同源性分别为99.31%和98.96%。     

Harpin-elicited hypersensitive cell death and pathogen resistance require the NDR1 and EDS1 genes

Plants sprayed with harpin, a bacterial protein that induces hypersensitive cell death (HCD), develop systemic acquired resistance (SAR) without macroscopic necrosis. HCD sometimes accompanies the development of resistance conferred by resistance (R) genes. In Arabidopsis, some R genes require one or both of the signalling components NDR1 and EDS1 for function. This study addresses whether HCD, NDR1 and EDS1 are required for induction of SAR by harpin. When Arabidopsis and tobacco leaves were sprayed with harpin, microscopic hypersensitive response (micro-HR) lesions developed. Systemic expression of PR genes and the development of resistance were accompanied by micro-HR, except in the ndr1-1 mutant, in which harpin induced micro-HR without the development of resistance or expression of the PR-1 gene. Cell death and resistance did not occur following treatment with harpin in plants that could not accumulate salicylic acid. Harpin also failed to induce resistance in Arabidopsis eds1-1 mutants. Therefore, harpin-induced resistance seems to develop concomitantly with cell death and resistance requires NDR1 and EDS1.     

Effect of organic treatments with calcium carbonate and bio-activator on quality of ‘Reinette’ apple cultivars

‘Reinette du Canada’ (RC) and ‘Reinette Grise du Canada’ (RG) apple (Malus × domestica Borkh) cultivars declared throughout the Community as Protected Designation of Origin ‘Manzana Reineta del Bierzo’ are severely affected by bitter-pit during storage. Pre-harvest treatments with calcium carbonate, authorized in organic production, and bio-activator Harpin protein were used to assess the effect on quality at harvest and during cold storage in both apple cultivars during 2007 and 2008. Bitter-pit at the end of storage was higher in ‘RC’ than in ‘RG’, due to the fact that K/Ca ratio in fruit was higher in ‘RC’. Harpin protein did not improve the quality of ‘Reinette’ apple cultivars. Calcium carbonate pre-harvest treatments were useful to decrease external and internal bitter-pit incidence of ‘Reinette’ apple cultivars after 90 days of storage, but differences at the end of storage were not significant. Therefore, calcium carbonate would be a useful product in organic production in order to decrease bitter-pit incidence in ‘Reinette’ apple cultivars during medium term storage.