酶联免疫吸附法在检测中的应用

酶联免疫吸附法(ELISA)是一种基本的酶免疫分析方法,起始于1971年Engval用碱性磷酸酯酶标记的3 gG测定1gG,随后这种方法得到迅速发展和不断完善.由于ELISA具有选择性好、灵敏度高、结果判断客观准确、实用性强等优点,目前已广泛应用于临床医学、生物学和分析化学等领域.……     

重组几丁质酶对采后葡萄贮藏过程中抗病效果的研究

以克瑞森无核葡萄品种为试材,采用重组几丁质酶对接种灰霉菌的葡萄果实进行处理,研究重组几丁质酶对葡萄果实腐烂的抑制及其诱导抗病作用。结果表明:在25℃条件下,重组几丁质酶处理能诱导采后葡萄的抗病性,不仅增加了葡萄果实中POD、PPO和PAL的酶活性,而且降低了丙二醛含量,诱导了果实中酚类物质含量的积累;在不同程度上延缓了葡萄的发病时间,降低了发病指数,贮藏至第6天时,对照的发病指数已达到100%,而重组几丁质酶处理的发病指数仅为47.8%。总的来看,重组几丁质酶处理可以提高葡萄采后抗病性,在贮藏初期抑制由灰霉引起的葡萄腐烂。     

天津市茄子灰霉病菌的抗药性检测

从天津市茄子保护地中分离出１２２个茄子灰霉病菌株中,多菌灵的抗性菌占４１．０％,且多为高抗菌株;多菌灵与乙霉威间表现负交互抗性关系;速克灵和扑海因的抗性菌比率为２７．９％,均为低抗水平,ＥＣ５０为２．３０１１￣２０．３８１０μｇ／ｍＬ,两种药剂存在交互抗性;对多菌灵、速克灵、扑海因均有抗性的菌株占供试菌株的２７．９％,均敏感的菌株占５９．０％,对多菌灵有抗性而对速克灵和扑海因敏感的菌株只占１３．１     

洛阳市番茄灰霉病菌对嘧霉胺的抗药性检测

为了明确洛阳市番茄灰霉病菌对苯胺基嘧啶类杀菌剂嘧霉胺的抗药性状况,2012—2013年从洛阳市不同保护地中采集番茄灰霉病病果或病叶,经单孢分离共获得番茄灰霉病菌54株。采用菌丝生长速率法测定了其对嘧霉胺的敏感性。结果显示,高抗菌株9株（16.67%）,中抗菌株19株（35.19%）,低抗菌株10株（18.52%）,敏感菌株16株（29.63%）。比较抗性水平不同菌株间的菌丝生长情况,发现菌丝生长速度和抗性水平无相关性。研究表明,洛阳市番茄灰霉病菌已对嘧霉胺产生不同水平的抗药性,且以中抗菌株为主,生产上应合理更换或轮换使用与嘧霉胺无交互抗性的药剂防治番茄灰霉病。     

草莓灰霉病菌对速克灵的抗性研究

速克灵对草莓灰霉病具有很好的防治效果,但长期、单一使用很容易使草莓灰霉病菌产生抗药性。为合理使用这类杀菌剂,延缓或防止抗药性的产生,笔者通过UV-C诱变草莓灰霉病菌敏感菌株CF1获得了对速克灵具有高抗性的突变株CF1-1和CF1-2,并对抗性突变株的生长量、产孢量和致病力进行了研究,结果表明:CF1-1与CF1-2对速克灵抗性程度高,抗性水平与敏感菌株相比分别增加了484.6倍和911.0倍。抗性菌株在无速克灵的培养基上菌丝生长较敏感菌株弱,产孢量也明显少于敏感菌株;但其引起草莓离体叶片发病较敏感菌株重,病斑大且边缘颜色深;薄片黄瓜测定结果也表明,在无速克灵的情况下,敏感菌株CF1引起发病较重;而在有药条件下CF1受到一定抑制,但抗性菌株仍能致黄瓜片发病。将抗性菌株连续培养7代后对速克灵的抗性仍是相对稳定的。     

Inheritance of resistance to Botrytis cinerea Pers. in Cicer arietinum L.

Summary Chickpea (Cicer arietinum L.) line ICC 1069 was selected as resistant parent after screening for resistance to grey mould (Botrytis cinerea Pers.) under artificial inoculation conditions. It was crossed with four high yielding susceptible varieties of chickpea. Crosses ICC 1069 × BGM 413 and ICC 1069 × BG 256 showed monogenic dominant resistance in ratio of 3R (resistant): 1S (susceptible). However, in crosses, ICC 1069 × BGM 419 and ICC 1069 × BGM 408, a ratio of 13S (susceptible) : 3R (resistant) was obtained indicating the presence of epistatic interaction. The results pointed towards the presence of a type of major gene resistance to grey mould in chickpea.     

The potential role of PR-8 gene of apple fruit in the mode of action of the yeast antagonist,Candida oleophila,in postharvest biocontrol of Botrytis cinerea

Several pathogenesis-related (PR) genes of apple have been cloned and their response to different pathogens has been studied. Different PR genes, however, may have a variable response depending on the specific organ or tissue as well as microbe. The objective of the current study was to characterize the expression of specific apple PR genes in fruit tissues in response to the antagonistic yeast, Candida oleophila (a common postharvest biocontrol agent), and the fungal pathogen, Botrytis cinerea (a major postharvest pathogen). Apple PR-5 and PR-8 gene expression was characterized in fruit in response to C. oleophila and B. cinerea. Results indicated that PR-8 expression was significantly elevated in response to both fungi. In contrast, neither C. oleophila nor B. cinerea treatment markedly affected PR-5 expression. The PR-8 gene was then synthesized and cloned into a Pichia pastoris expression system to study the antifungal activity of the PR-8 protein against B. cinerea both in vitro and in vivo. Collectively, the PR-8 gene of apple is associated with the response to B. cinerea infection, and may play a role in the mechanism by which C. oleophila effectively inhibits B. cinerea disease in apple fruit, namely by the induction of this specific host PR gene.     

春季大棚番茄灰霉病发生规律及影响因子分析

为探索春季大棚灰霉病的生态调控技术,采用定点观察的方法,开展灰霉病发生规律系统监测。结果表明,灰霉病的发生大致可分为3个时期,即4月上中旬的快速增长期、4月下旬至5月上旬的稳定增长期和5月下旬以后的缓慢增长期。通过对棚内温度、湿度、病情基数等因子分析,结果表明灰霉病的发病率与病情基数、大棚日平均温度、大棚日平均相对湿度呈极显著的正相关。因而生产上可采用摘除病叶、调控棚内温湿度等生态调控措施来控制病情的发生扩展和蔓延。     

Heredity of Resistance to Botrytis cinerea in Sunflower (Helianthus annuus) Measured by Artificial Infections

Genetic Variability for Some Brewing Quality Characters in Barley Malting and brewing quality of six different barley genotypes and their F1 hybrids were investigated in a diallel cross programme. Plants were grown in pots in a randomized block design with four replications. General and specific combining abilities were determined for 1000 grain weight, percentage of proteins of the grain, percentage of water uptake, and, after micromalting, for β-glucanase and α-amylase activities. Significant general and specific combining abilities were observed for all the traits studied. 1000 grain weight, percentage of proteins and water uptake have been found to be essentially controlled by genetic additive effects, while enzymatic activities of malt have been found to be mostly controlled by dominance effects. ‘Karl’ has interesting characteristics for all traits and general combining abilities, except for 1000 grain weight, and has been introduced in a three-way cross programme with ‘Plaisant’ and ‘Clarine’, also showing interesting values for malting quality and productivity.     

N-3-羰基十四烷酰基高丝氨酸内酯诱导番茄对灰霉病抗性机制初探

为了探究细菌群体感应(QS)信号分子N-酰基高丝氨酸内酯(AHLs)在植物上诱导对于真菌病害的抵抗能力。以番茄为试验材料,以灰霉为病原指示菌,通过不同侧链长度和侧链修饰的AHLs信号分子预处理后接种病原菌,发现长链信号分子N-3-羰基十四烷酰基高丝氨酸内酯(3OC14-HSL)对灰霉有最佳的抗性效果。实时荧光定量PCR检测抗病基因发现,与未处理对照组相比,信号分子3OC14-HSL预处理能显著诱导接种灰霉病后的番茄体内与茉莉酸(JA)信号通路相应基因PI1、PI2上调;而与水杨酸信号(SA)通路相关的NPR1基因显著下调,PR1基因则没有显著变化。进一步通过DAB染色、过氧化氢含量测定以及过氧化物酶(POD)和超氧化物歧化酶(SOD)活性检测发现,3OC14-HSL预处理相比未处理对照组使得番茄产生大量活性氧,且可以保持较高的POD、SOD活性。这些结果表明,长链信号分子N-3-羰基十四烷酰基高丝氨酸内酯可以诱导番茄对灰霉的抗性,而且该抗性效果可能与茉莉酸信号路径相关。     

Development of methods using phytoalexin (resveratrol) assessment as a selection criterion to screen grapevine in vitro cultures for resistance to grey mould (Botrytis cinerea)

Summary The purpose of this research was, firstly to determine the ability of grapevine in vitro cultures to synthesize resveratrol, a stilbene-type phytoalexin that is considered to be a good marker for resistance of grapevines to Botrytis cinerea, the causal organism for grey mould. Secondly, this study sought to establish the relationship between phytoalexin production potential and resistance to Botrytis cinerea in grapevines. In this aim, resveratrol production was assessed in 13 Vitis species or cultivars. A good correlation appeared between resveratrol production by grapevine in vitro cultures and grey mould resistance except for two Vitis spp. for which no correlation was observed, thus suggesting that resistance of grapevines may sometimes be associated with factors other than stilbene phytoalexins. In view of the results obtained, the potential use of resveratrol induction and of in vitro methods as a tool for screening grapevines for resistance to B. cinerea was discussed.