稻瘟菌诱导的稻叶脂氧合酶的纯化及部分氨基酸序列测定

 作者对已报道的稻瘟菌诱导稻叶脂氧合酶CM-LOX 1和CM-LOX 2的纯化方法作了进一步改进和完善。改进后的纯化方法可缩短该脂氧合酶的纯化时间,并提高纯化倍数。利用改进的方法,作者从20 g非亲和性稻瘟菌小种接种稻叶中获得了电泳纯的酶蛋白CM-L OX 1 133 μ g和CM-LOX 2 208μg。在此基础上,采用特异性的蛋白酶endoproteinase Lys-C和溴化氰对该酶蛋白进行了裂解,回收其肽段,得到了CM-LOX 1的3个不同肽段共30个氨基酸和CM-L OX 2的7个不同肽段共87个氨基酸的序列,为最终克隆这2个酶的基因奠定了基础。     

抗稻瘿蚊分子育种研究进展与利用

综述了近年来在抗稻瘿蚊基因的分子标记和分子标记辅助选择育种方面的研究进展及其利用。     

Partial resistance to neck blast influenced by stage of panicle development and rice genotype

Summary Neck nodes of eight rice genotypes were inoculated with a virulent isolate of the blast pathogen at four slightly different, increasingly older stages of panicle development shortly after flowering. Resistance to infection as well as resistance to growth of the pathogen after infection was assessed. Significant differences between genotypes were found for both components of resistance. Resistance to growth of the pathogen sharply increased with aging of the neck node as was indicated by a large reduction of the length of the lesions on the culm, but resistance to infection did not change much. The lesion length was closely related to yield loss. A small delay in time of infection can lead to a large decrease in yield reduction. Meaningful comparison of partial resistance to neck blast between genotypes therefore requires infection at an identical stage of panicle development. In the field, where panicles are in different stages of development, selection of genotypes that do show the symptoms of neck blast together with relatively little yield loss in infected panicles is recommended.     

曲霉型豆豉用高产蛋白酶菌种的选育

从曲霉型阳江豆豉曲中筛选出5株菌落形态不同的曲霉(Aspergillus spp.),通过纯种发酵制备豆豉曲,测定了豆豉曲中碱性蛋白酶和中性蛋白酶的活性,从中筛选出1株蛋白酶活性较高的菌种,并对该曲霉进行ITS r DNA序列分析,发现该曲霉与Aspergillus oryzae strain P6B2(JX429926)的同源性最高,故鉴定为米曲霉(Aspergillus oryzae).以该米曲霉为出发菌株,通过紫外线诱变和硫酸二乙酯化学复合诱变处理,选育得到优良高产蛋白酶活性的突变株,其中性蛋白酶酶     

Expression and homology modelling of sterol 14α‐demethylase of Magnaporthe grisea and its interaction with azoles

BACKGROUND: Magnaporthe grisea (Hebert) ME Barr infection is one of the most serious diseases for cultivated rice in the world. Sterol 14α‐demethylase (CYP51) is an important drug target for microbial pathogenic infections. To exploit specific and effective fungicides for M. grisea better, the authors have analysed the characteristics of interaction between sterol 14α‐demethylase from M. grisea (MGCYP51) and azoles. MGCYP51 with truncation of N‐terminal residues was cloned and expressed in E. coli, difference binding spectra of MGCYP51 induced by addition of four commercial azoles were determined and molecular modelling of MGCYP51 based on the crystal structure of Mycobacterium tuberculosis Lehmann & Newman and docking with the azoles were performed. RESULTS: The affinity of the azoles for MGCYP51 was positively correlated with their hydrophobicity. Amino acid residues Tyr112, Phe120, Phe220, His308 and Phe497 of MGCYP51, forming a large hydrophobic cavity, are the key residues interacting with azole fungicides. Furthermore, Phe220 and Phe497 are fungus and species specific respectively. CONCLUSION: The results suggest that the more potent azole fungicides for MGCYP51 should possess more hydrophobic groups interacting with residues Phe220 and Phe497. Copyright © 2009 Society of Chemical Industry     

甲酸乙酯对米象不同虫态的熏蒸作用

针对储粮害虫抗药性日趋严重、粮食熏蒸剂品种越来越少的现状,采用密闭熏蒸法,系统研究甲酸乙酯（EtF）在不同条件下对米象Sitophilus oryzae（L.）卵、低龄幼虫、高龄幼虫、蛹以及成虫的致死作用.在25℃条件下以40和45μL/L的药剂浓度处理米象成虫12h,其校正死亡率分别为90.541%和99.324%;以30μL/L的浓度在19、25和31℃下处理24h,校正死亡率分别为83.108%、62%和42%,表明甲酸乙酯具有良好的速效性,且在相对低温时比在相对高温时熏蒸效果好.在20、25和30℃条件下,熏蒸处理米象成虫24h,其LC50分别为25.094、28.650和30.875μL/L;处理48 h时则分别为23.101、25.805和27.316μL/L.30g/m^3的甲酸乙酯熏蒸处理米象卵、低龄幼虫、高龄幼虫和蛹24 h,对应死亡率为96.930%、99.333%、97.768%和93.386%,表明甲酸乙酯对米象低龄幼虫的熏蒸效果最好,蛹和卵对甲酸乙酯具有较高的耐药性.     

AtEDR1介导拟南芥对稻瘟菌的抗性(英文)

 稻瘟菌(Magnaporthe oryzae)是一种半腐生病原菌,该病原菌可以侵染水稻,产生稻瘟病。对于水稻与稻瘟菌的互作机制,人们已经进行了大量的研究。然而,鉴于水稻基因组相对较大且稻瘟菌生理小种众多,致使研究进展缓慢。本研究通过将稻瘟菌生理小种Y34侵染拟南芥生态型Col-0,发现Y34可以感染Col-0,从而建立Y34-拟南芥互作模型,用以探究Y34与抗白粉突变体edr1的关系。结果显示,edr1比Col-0对于稻瘟菌更敏感。之前有研究表明,降低SA含量的突变体(pad4,sid2,nim1)均可抑制edr1相关表型。接种Y34于edr1与pad4、sid2、nim1形成的双基因突变体发现,所有双突变体的感病性比edr1单突均明显降低。推测EDR1可能在拟南芥抗稻瘟菌方面起一定正调控作用,且其作用依赖于SA途径的相关基因。     

不同杀菌剂对草坪草病原菌毒力的作用测定

采用生长速率法测定了不同杀菌剂对3种主要引起坪草病害的褐斑病菌、腐霉枯萎菌、夏季斑枯病菌的毒力。结果表明,烯唑醇、甲基硫菌灵、代森锰锌、咪鲜胺对立枯丝核菌的EC50值分别为0.078 3,5.968 4,7.181 2和11.386 8 mg/L,以烯唑醇对立枯丝核菌的毒力最高。烯唑醇、咪鲜胺、代森锰锌、甲基硫菌灵对夏季斑枯病菌的EC50值分别为0.016 2,0.554 4,6.035 3,1 218.497 8 mg/L,以烯唑醇抑菌效果显著优于其它3种杀菌剂。采用菌丝干重测定法测定了阿米西达、霜脲氰、霜克、咪鲜胺、代森锰锌对腐霉枯萎病菌的毒力,其EC50值依次为0.053 3,7.837 4,13.310 7,19.715 1,29.771 5 mg/L,以阿米西达的抑菌效果最好。     

水稻中对不亲和白叶枯病菌株专一识别的多肽

 经蛋白质双向电泳分析，发现水稻IR26、IR1545及TN1各有一个多肽能与不亲和的白叶枯病菌株产生特异性作用而消失，但与亲和菌株相互不发生作用。成株期抗病品种南粳l5在成株期有与不亲和菌株作用的多肽存在．而在苗期则未观察到这一多肽。对全期抗病品种IR26分析表明，它无论在成株期还是苗期均出现一个与不亲和菌株专一作用的多肽。经典遗传学分析认为有三个抗白叶枯病基因的DV85，则观察到DV85有两个以上的菌株特异性多肽。由于选些多肽表现出菌株的专一性及其它表达生理时期均与该品种的田间抗病表型一致，同时多肽的数目也与经典遗传学分析结果类似，推测这些多肽可能是抗病基因的产物。     

湖南省稻瘟病菌无毒基因鉴定

【目的】了解湖南省稻瘟病菌无毒基因组成,为培育水稻新品种及合理布局抗稻瘟病品种提供参考依据。【方法】采用2014年分离自湖南桃江病圃丽江新团黑谷(LTH)上的120个稻瘟病菌单孢菌株室内离体针刺接种于24个已知抗瘟基因的水稻近等基因系(NILs)上,对供试菌株进行无毒基因鉴定。【结果】120个稻瘟病菌含有全部24个无毒基因,在24个无毒基因中,出现频率高于40.00%的有Avr-Pita和Avr-Pish;出现频率在30.00%~40.00%的无毒基因有Avr-Piz、Avr-Piz5、Avr-Pi3和Avr-Pi9;其余无毒基因出现频率均较低,特别是Avr-Piks、Avr-Pikp、Avr-Pib、Avr-Pi12和Avr-Pi11出现频率均低于10.00%。【结论】含抗性基因Pi-ta和Pi-sh的水稻品种可在湖南省推广利用;含抗性基因Pi-z、Pi-z5、 Pi-3和Pi-9的水稻品种在湖南推广时应慎重考虑;含抗性基因Pi-ks、Pi-kp、Pi-b、Pi-12和Pi-11的水稻品种不宜在湖南省推广利用。