纤维素产氢菌的分离及不同秸秆处理方法的研究

为了增强生物方法对秸秆的利用能力,将秸秆较好地转化成氢能,本试验利用改良的Hungate厌氧滚管法及滤纸崩解法,从已得菌系JYB中,筛选得到1株能较好地降解纤维素产氢的细菌,并利用水热法、汽爆法等6种方法对秸秆进行预处理,以增强秸秆的产氢效果。结果表明：筛选得到菌株JYB-13,该菌可利用大量碳源产氢,产氢的最适条件为35℃,pH值8.5。对秸秆进行预处理,由产氢效果得知,该菌对2%硫酸处理后的秸秆降解效果最好,秸秆的降解率为53.1%,氢气含量达到37.2%。分析发酵前后秸秆的成分,可知该菌可以较好地利用纤维素和半纤维素,对木质素利用较少。     

Evaluation of Dutch elm disease susceptibility and pathogenicity of Ophiostoma ulmi using micropropagated elm shoots

Elm (Ulmus sp.) shoots from in vitro micropropagated plants were tested with Ophiostoma ulmi culture filtrates, at different dilutions to determine the susceptibility of different host clones to Dutch elm disease (DED). ‘Commelin’, a susceptible Dutch hybrid, showed a high wilting index but that of American elm was higher. Although U. pumila × japonica (Sapporo gold 2) was the most tolerant to spore inoculation, its wilting index was as high as that of ‘Commelin’. The absorption capacity and wilting index of the elm shoots showed consistent correlation to isolate effectiveness (as determined by inoculation experiments), to production of toxin (cerato-ulmin) and to filtrate dilution. This method could be used to assess the effectiveness of the different O. ulmi isolates and for the detection of elm genotypes that are highly susceptible to Dutch elm disease.     

耐酸性α-淀粉酶产生菌发酵条件的优化

试验对从淀粉厂附近的土壤中筛选的产耐酸性α-淀粉酶的菌株发酵条件进行了考察,确定最佳发酵条件为pH 4.5、碳源为可溶性淀粉、氮源为柠檬酸二氢铵、培养温度为35 ℃、接种量为2 mL。      

凝固型黑豆酸奶工艺的研究

将保加利亚乳杆菌和嗜热链球菌在牛奶与黑豆浆培养液中进行连续传代驯化,使其逐步适应黑豆奶的发酵环境,再用驯化后的菌种生产黑豆酸奶。同时对黑豆乳去腥、豆浆浓度进行了探讨,并确定了主要工艺参数。     

秸秆复合有机酿热物对大棚土壤环境的影响

为解决黑龙江省早春大棚土壤温度低及设施土壤环境恶化问题,以‘千禧’番茄为材料,研究秸秆+牛粪(NGH)、秸秆+马粪(MGH)、秸秆+羊粪(YGH)、单一秸秆(JG),以未进行反应堆技术为对照(CK),对早春大棚‘千禧’番茄土壤温度、土壤理化性状、酶活性及微生物群落结构的影响。结果表明,与对照相比,各处理降低了土壤pH,提高了土壤温度、土壤EC、碱解氮、速效钾、有机质含量及土壤蔗糖酶、脲酶、磷酸酶、过氧化氢酶活性。经过秸秆生物反应堆处理的20 cm土层温度有明显的提升,其中秸秆+羊粪处理在反应启动后提升土壤温度效果最好;秸秆+羊粪(YGH)处理提高了土壤速效磷含量,是对照的2倍;秸秆+羊粪(YGH)处理土壤中4种酶活性均处于最高水平,分别比对照提高了91.9%、220%、18.5%、7.3%。各处理微生物多样性指数均显著高于对照,变化规律一致为秸秆+羊粪(YGH)>秸秆+牛粪(NGH)>秸秆+马粪(MGH)>秸秆(JG)>对照(CK);秸秆+羊粪(YGH)及秸秆+牛粪(NGH)处理对糖类、氨基酸类、酯类、醇类、胺类、酸类6类碳源的利用能力最强,显著高于其他处理和对照。应用主成分分析和聚类分析方法将大棚番茄土壤分为4个等级,秸秆+羊粪(YGH)处理为第一级,秸秆+牛粪(NGH)处理为第二级,秸秆+马粪(MGH)及单一秸秆(JG)处理为第三级,对照(CK)为第四级。应用秸秆复合有机酿热物生物反应堆促进了土壤环境的改善,以秸秆+羊粪(YGH)作为复合有机酿热物效果最佳。     

转基因球孢白僵菌发酵产物对野生菌株的增效作用

研究发现,超量表达类枯草杆菌蛋白酶基因CDEP-1的球孢白僵菌菌株Bb0062-15-4-CDEP-1的发酵产物对野生菌株具有明显的杀桃蚜增效作用。进一步分析增效作用与目的基因表达产物关系的结果显示,加入CDEP-1酶活为44.3±0.6U/μL的发酵产物,可使野生菌株对桃蚜的致死中浓度降低4倍以上,共毒系数(CTC)达404.6。但发酵产物经121℃处理20min,增效作用完全丧失。用饱和硫酸铵沉淀蛋白质分别将透析后的重溶蛋白质和上清液用于增效作用测定,结果证明增效成分主要是蛋白类物质。将梯度稀释的发酵产物与浓度为1×107孢子/mL的野生菌株孢子悬浮液混合进行生物测定,发现各处理的致死中时与CDEP-1的活性呈负相关,相关系数为-0.8946。由此推断,发酵产物中的增效物质主要是目的基因表达产物CDEP-1。     

Resistance to late blight (Phytophthora infestans (Mont.) de Bary) of potato plants regenerated fromin vitro selected calli

Summary Cells of seven potato cultivars were selectedin vitro with culture filtrate (CF) ofPhytophthora infestans (Mont.) de Bary. Regenerated plants were tested for late blight resistance. The aim of the study was to check the efficacy of CF in the selection of potatoes for resistance to late blight and to evaluate the effects of additive factors on general resistance. One selection cycle, applied to a cell culture system, selected cells resistant to toxic metabolites of CF.In vivo screening of clones regenerated from selected cells was done in two steps: on whole plants, assessing foliage late blight, and on detached leaves, assessing single factors of horizontal resistance. In general, the frequency of resistant variants selected with CF did not differ from that of resistant somaclonal variants. Nevertheless, high concentrations of CF in the growth medium seemed to induce an improvement of some partial resistance factors as compared with the source plants.     

Responses of nematode‐community structure in turfgrass soil to microbial filtrates from municipal–solid waste compost

Actinomycetes, Bacillus subtilis, and Bacillus thuringiensis were isolated from municipal–solid waste (MSW) compost, and different microbial liquid filtrates (MLF) were prepared. Sterile culture media with no microbes were used as their controls. The effects of MLF on soil nematode communities were examined in pot‐grown Festuca arundinacea Schreb. Fifteen genera of nematodes in background soil were identified, of which Helicotylenchus and Rotylenchus were dominant. The inoculation of MLF strongly affected the abundance and community structure of soil nematodes. Compared with their controls, lower total nematode numbers following MLF incorporation were found. Actinomycetes inoculation changed community structure of soil nematodes, transforming the dominant genera from Helicotylenchus and Rotylenchus into Cephalobus, Chiloplacus, and Aphelenchus. Actinomycetes incorporation resulted in a significant decrease of plant‐parasitic nematodes relative to control pots. Only plant‐parasitic and omnivorous‐predatory nematodes were found in treatments following B. subtilis inoculation, and Helicotylenchus, Rotylenchus were dominant genera with relative abundance of 76.2% and 14.3%, respectively. Although the dominant genera were still Helicotylenchus and Rotylenchus, B. thuringiensis inoculation led to a marked decrease in populations of plant‐parasitic nematodes and an increase in populations of fungivorous and bacterivorous nematodes relative to control. Shannon's diversity index (H′), evenness index (J′), richness index (SR), and Wasilewska index (WI) in pots treated with actinomycetes and B. thuringiensis filtrates were significantly higher than those of their controls, whereas significant lower dominance index (λ) in actinomycetes and B. thuringiensis treatments was observed than their controls. Plant growth was improved in the treatments inoculated with three microbes. The findings highlight that actinomycetes can most effectively suppress plant‐parasitic nematodes, increase community diversity, evenness, and richness, thus improving soil environment for turf growth.     

油菜内生细菌yc8发酵产物特性初探

实验从山西太谷、运城采集的油菜中分离得到40株植物内生细菌,经过初步筛选得到具有生防潜力的菌株yc8,该菌株发酵产物对多种植物病原真菌有强烈的抑制作用,并对其发酵产物的特性进行了进一步的研究。结果表明:yc8在发酵培养20 h时,菌液的吸光值达最大值1.68,其蛋白质浓度也达到最大值0.478 mg/mL;在发酵培养48 h时,其发酵滤液对向日葵菌核病菌的抑菌率达到最大值,为83.0%。发酵液对热和紫外线具有较强的稳定性;在pH6~9的溶液里抑菌活性较稳定;在极酸极碱条件下抑菌活性丧失;对氯仿的稳定性较差。     

煤矸石淋滤液对小麦种子及幼苗的生态毒性研究

以山西省农业科学院提供的小麦品种长麦6135为研究对象,采用室内培养方法和柱状淋溶装置,模拟大气降水,用5个不同浓度的煤矸石淋滤液处理小麦种子,分析其对小麦发芽势、发芽率、叶绿素含量和抗氧化酶活力等的影响。结果表明：煤矸石淋滤液对小麦种子的发芽有一定抑制作用,浓度越高,抑制作用越显著,煤矸石淋滤液浓度为720．8 mg／L 时小麦发芽势和发芽率均最低,分别为65．6％、69．0％。煤矸石淋滤液浓度为324．5～720．8 mg／L 时,小麦芽长与苗高均显著低于对照组,当浓度为451．5～720．8 mg／L 时,小麦苗质量显著低于对照组;叶绿素含量与煤矸石淋滤液浓度呈线性负相关。不同煤矸石淋滤液浓度的小麦幼苗 POD 活性差异性显著,而 SOD 和CAT 活性差异性极显著;不同部位的 POD 和 SOD 活性差异极显著;不同时间 POD 和 CAT 活性差异极显著。因此,煤矸石淋滤液对小麦种子发芽和幼苗生长具有一定的生态毒性。