Antibacterial mechanism of cow manure biogasslurry on Rhizoctonia solani

为探讨牛粪发酵沼液的抑菌机理,以立枯丝核菌为研究对象,进行了试验研究。采用平皿培养方法：培养基表面分别均匀涂布1 mL 5种不同处理的沼液,即不作处理沼液、沼液灭菌、离心上清液、无菌膜滤液和菌体溶液,中心接种立枯丝核菌并于生化培养箱中28℃培养96 h,每12 h测量立枯菌落的生长直径以计算抑制率,同时借助分子生物学手段鉴定了沼液中起抑制作用的微生物。试验结果表明菌体溶液和不作处理沼液对立枯的抑制效果较好,抑制率最高可达74%和53%,离心上清液的抑制效果次之,无菌膜滤液和沼液灭菌处理的抑制作用较弱,一般在20%以下;对沼液中的微生物菌群进行了分离、纯化与筛选,获16株菌种,并与立枯丝核菌进行对峙培养试验,其中nfd-9和nfd-16具有较强抑制作用,其他菌株几乎没有抑制作用。将nfd-9、nfd-16与立枯对峙交界面处进行了扫描电镜观察,结果显示：这2株微生物通过竞争、拮抗和重寄生等作用来抑制立枯丝核菌的生长,从而到达防止病害的目的。经分子生物学技术鉴定,确定了nfd-9和nfd-16分别为米根霉菌（Rhizopus oryzae）和哈茨木霉（Trichoderma harzianum）。研究结果为进一步了解沼液抑制真菌的机理提供一定的理论参考。     

Control of post‐harvest decay of apples by pre‐harvest and post‐harvest application of ammonium molybdate

Ammonium molybdate was tested as a potential fungicide for use in apples (cv Golden Delicious) against blue and grey mould, important post‐harvest diseases of pome fruits. In tests in vivo at 20 °C, ammonium molybdate (15 mM ) reduced lesion diameters of Penicillium expansum, Botrytis cinerea and Rhizopus stolonifer by 84%, 88% and 100% respectively. When apples treated with ammonium molybdate were stored at 1 °C for three months, a significant reduction in severity and incidence of P expansum and B cinerea was observed in both years of study (1998 and 1999). In the second year of the experiment the reduction in disease severity was greater than 88% for both pathogens, and the level of control was similar to, or greater than, that observed with the fungicide imazalil. When ammonium molybdate was applied as a pre‐harvest treatment, a significant reduction in blue mould decay was observed after three months in cold storage. In vitro, ammonium molybdate greatly inhibited spore germination of P expansum and B cinerea, although better inhibition was obtained against grey mould. Ammonium dimolybdate, sodium molybdate and potassium molybdate were also tested in vitro in comparison with ammonium molybdate as inhibitors of spore germination, but only ammonium molybdate inhibited spore germination by more than 50%. © 2001 Society of Chemical Industry     

同步辐射软X射线Nk对产L-乳酸米根霉的诱变效应

采用同步辐射软X射线Nk对米根霉AS 3.3461菌株进行辐射诱变,研究其对米根霉致死、菌落形态、种子培养以及发酵培养等特性方面的影响。结果表明,致死率曲线呈现为典型的马鞍型曲线,辐射剂量为0.72 kGy时菌株致死率最低;软X射线Nk辐照对米根霉菌落形态有一定影响;种子培养特性和发酵培养特性的诱变前后对比表明,种子延滞期缩短了3h,突变株N-15的L-乳酸产量提高了25.6%,LDH活性提高了95.8%,ADH的活性降低了77.7%。     

复合诱变筛选丙烯醇抗性的富马酸高产米根霉菌株

[目的]筛选具有丙烯醇抗性的富马酸高产米根霉菌株。[方法]以溴甲酚绿与丙烯醇平板作为双初筛标准,建立代谢调控选育米根霉富马酸高产菌株的方法。[结果]利用紫外诱变对米根霉孢子的最佳诱变条件分别为紫外灯30 W,垂直照射距离30 cm,诱变时间1 min;化学诱变剂亚硝酸浓度0.1 mol/L,诱变时间10 min。优化条件下,通过紫外和化学的复合诱变获得了1株产量高、稳定性好的米根霉丙烯醇抗性突变株ME-F12,富马酸产量达到50.39 g/L,与原始菌株相比提高23%,连续传代5次后,产量无明显降低,具有较高稳定性,副产物乙醇浓度只有1.87 g/L,明显低于原始菌株。[结论]为微生物发酵法生产富马酸的工业化奠定了扎实的基础。     

广州附近的草莓病害

 草莓在广州附近的主要病害为轮斑病(Phomopsis obscurans)、灰霉病(Botrytis cinerea)、灰斑病(Cercospora fagarina sp.nov.)、叶霉病(Cladosporium cladosporioides)、基腐病(Rhizoctonia solani)、软腐病(Rhizopus stolonifer,R.sexualis)和褐斑病(Pestalotiopsis adusta)等7种,尤以第一、二种危害严重,Cercospora fragarina是个新种,Rhizopus sexualis是国内新纪录,草莓在广州附近虽试种成功,今后扩大栽培,不可不注意病害的防治。     

温度对葡萄烂果病病原菌(黑根霉)的影响

该文研究了温度对葡萄烂果病病原菌(黑根霉,Rhizopus nigricans Ehrenb)的影响,结果表明:葡萄烂果病病原菌(黑根霉)菌丝发育的初始温度为5℃,最适生长温度为25～30℃。致死,40℃下需要48h,45℃下需24h。孢子囊具有很强的抗逆性。     

同步辐射CNOk软X射线对米根霉诱变效果比较分析

同步辐射软X射线具有能谱宽且连续、精确可调、单色性好、可精确计算辐照剂量等特点，可提供生物分子所具有的C，N，O和k层电子吸收边波长的射线，在生物学领域有着广泛的用途。采用同步辐射2.3，3．2和4．4nm波长软X射线对米根霉3．819孢子进行辐射诱变，研究抽真空时间、不同吸收边射线、辐照剂量与诱变效果的关系。结果表明，抽真空时间对米根霉孢子的存活率有很大影响，时间越短存活率越高。YPD平板筛选结果显示：Ck软X射线诱变致死率最高，筛选得到的突变株最少，主要产生在高剂量区；Nk软X射线的效果较好，突变株产生在低剂量区；Ok诱变致死率最低，筛选得到的突变株最多，在不同剂量范围内均有产生。大平皿点样得到Nk软X射线诱变后的11号突变株H/C比值最大，其发酵液经高效液相色谱测定产物几乎为乳酸。     

Effects of the yeast Pichia guilliermondii against Rhizopus nigricans on tomato fruit

The yeast Pichia guilliermondii was examined for its ability to control Rhizopus nigricans on tomato fruit during storage, and in order to highlight the reason for biocontrol, a possible mode of action is discussed. Results showed that autoclaved yeast culture and culture filtrate had no effect on controlling the postharvest disease caused by R. nigricans, although inoculation of P. guilliermondii prior to R. nigricans resulted in enhanced biocontrol efficacy. Moreover, rapid colonization of the yeast on wound sites was observed during the initial 3 days at 20 °C, and then the population stabilized for the remaining 4 days. This phenomenon indicated that at room temperature, P. guilliermondii could acclimatize itself to the environment of tomato fruit wounds and occupy the living space quickly. The results indicate that P. guilliermondii did not produce an antifungal substance, however, competition for nutrients and space on wounds appeared to play a role in the activity of the biocontrol and could be one of the mechanisms. In addition, the fruit inoculated with P. guilliermondii demonstrated changes in peroxidase (POD), polyphenoloxidase (PPO), superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), chitinase (CHI) and β-1,3-glucanase activities, all of which were correlated with the onset of induced resistance. This result suggests that tomato fruit is capable of responding to the yeast P. guilliermondii, which could activate defensive enzymes and thereby induce host disease resistance.     

拟康氏木霉和黑根霉混合发酵对苹果链格孢的防治

研究黑根霉和拟康氏木霉混合发酵液对苹果链格孢菌的生物防治效果,探索开发为新型绿色环保生物防治剂。对2种生防微生物进行形态学鉴定和ITS鉴定;生长速率法测定混合发酵液的抑菌活性;在烟台莱阳进行田间实验对发病较为严重的果园进行为期一年的多次灌根与喷施处理,测定其防治效果并测定其对苹果叶片SOD、POD等防御相关酶的活性。研究结果表明,黑根霉和拟康氏木霉混合发酵可显著提高对病原微生物链格孢菌的抑菌能力。平板抑菌实验结果表明,混合发酵液对链格孢菌的抑菌效果达到了66.73%,显著高于黑根霉和拟康氏木霉单独发酵液的51.23%、17.28%;田间实验发现,混合发酵液处理后苹果斑点落叶病发病率与对照组相比显著下降,相对防效达到42.46%;对其叶片防御酶测定结果表明,混合发酵液处理后,苹果叶片的防御酶SOD、POD、PAL活性均有50%以上的提高。黑根霉和拟康氏木霉混合发酵液对苹果链格孢菌具有显著的抑制作用,能有效防治苹果斑点落叶病,提高苹果抗病能力,为新型生防制剂的开发提供了参考。     

同步辐射软X射线Nk对产L-乳酸米根霉的诱变效应

采用同步辐射软X射线Nk对米根霉AS 3.3461菌株进行辐射诱变,研究其对米根霉致死、菌落形态、种子培养以及发酵培养等特性方面的影响。结果表明,致死率曲线呈现为典型的马鞍型曲线,辐射剂量为0.72 kGy时菌株致死率最低;软X射线Nk辐照对米根霉菌落形态有一定影响;种子培养特性和发酵培养特性的诱变前后对比表明,种子延滞期缩短了3h,突变株N-15的L-乳酸产量提高了25.6%,LDH活性提高了95.8%,ADH的活性降低了77.7%。