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     

Evaluation of chitosan and alginate immobilized Methylobacterium oryzae CBMB20 on tomato plant growth

Methylobacterium oryzae CBMB20, a promising plant growth promoting bacteria (PGPB) and a biocontrol agent, was immobilized in different formulations such as wet chitosan, dry chitosan, wet alginate and dry alginate and were tested for tomato plant growth promotion. Chitosan solution (1.5%) with pH 5.5–6.0 and 90 min contact time was found optimal for immobilization. The chitosan formulations showed better entrapment efficiency and good degradability resistance apart from slow release of cells under prolonged incubation. Survivability of bacteria (80%) was observed in wet chitosan formulation even after 90 days of storage at 4°C. The spermosphere survival of bacteria was high in both dry and wet chitosan formulations applied soils even after 21 days under greenhouse conditions. While the alginate formulation degraded fully, partial degradation of chitosan formulation was observed even after 30 days, indicating its ability to support the survival of M. oryzae CBMB20 in soil. Plants inoculated with wet chitosan formulation registered 1.3 fold increase in the shoot and root length and dry weight compared to other treatments. Hence, chitosan formulation supporting better plant growth compared to alginate will be a better carrier for taking bacteria to the plant rhizosphere and thereby promote plant growth.     

香蕉茎叶粉固态发酵条件优化及鹅对其养分利用率的研究

本试验旨在研究应用固态发酵技术改善香蕉茎叶粉养分组成的工艺参数及鹅对其养分的利用率。选择米曲霉和产朊假丝酵母作为发酵菌种,逐步探究5个因素(单菌种发酵、硫酸铵添加量、米曲霉和产朊假丝酵母接种比例、混合菌液接种量)对发酵后香蕉茎叶粉蛋白质含量的影响。利用正交设计筛选最优发酵温度、底物水分和发酵时间的组合。结果显示:米曲霉和产朊假丝酵母单独发酵均能显著或极显著提高发酵后香蕉茎叶粉的粗蛋白质含量(P0.05或P0.01)。添加硫酸铵能显著或极显著提高发酵后香蕉茎叶粉的真蛋白质含量(P0.05或P0.01),其中添加2%硫酸铵组的真蛋白质含量最高。米曲霉∶产朊假丝酵母接种比例为2∶1的处理发酵后香蕉茎叶粉的蛋白质净增加量显著或极显著高于比例为1∶1、1∶3和3∶2的处理(P0.05或P0.01)。正交试验结果显示,以4%的接种量、接种比例为2∶1(米曲霉∶产朊假丝酵母)、2%的硫酸铵添加量,在基质水分为50%,30℃的环境下发酵4 d效果最佳。经过该工艺发酵后的香蕉茎叶粉粗蛋白质含量提高了33.82%,氨基酸分析结果显示,除赖氨酸和精氨酸外其余的15种氨基酸含量均有不同程度的提高。马冈鹅的代谢试验结果显示,发酵后香蕉茎叶粉中的粗蛋白质利用率提高了52.66%,极显著高于发酵前(P0.01);此外,代谢能和能量利用率也都略有提高(P0.05)。由此可见,经过该发酵工艺发酵后的香蕉茎叶粉营养价值不仅得到了改善,也促进了鹅对其养分的消化吸收。     

小麦籽粒营养物质和次生物质含量与其对玉米象抗性的关系

为明确生产栽培小麦品种对玉米象Sitophilus zeamais(Motschulsky)的抗性机制,采用选择性和非选择性试验法系统评估了9个小麦品种对玉米象的抗性,并分析了小麦籽粒营养物质和次生物质含量与其抗性的关系。结果显示,不同小麦品种对玉米象的抗性存在显著差异,其中陕麦139和西农88的不选择性和抗生性较弱,小偃22、周麦16和周麦11较强;玉米象成虫对西农165选择性最强,虫口数为152.00头,而卵至子代成虫发育历期最长,为35.06 d,子代成虫数、重量损失率和敏感系数均较低,分别为19.67头、5.37%和8.30;对周麦18选择性最差,敏感性居中。玉米象成虫对不同小麦品种的选择性与各物质含量,以及总酚、蛋白质含量与各抗性指标均无显著相关性,但阿魏酸含量与子代成虫数、敏感系数及重量损失率显著负相关;可溶性糖含量与子代成虫数、敏感系数及重量损失率显著正相关,与卵至子代成虫发育历期显著负相关。表明小麦品种对玉米象的抗生性与其籽粒中阿魏酸和可溶性糖含量相关,阿魏酸含量越高、可溶性糖含量越低,小麦对玉米象的抗性越强。     

水稻白叶枯病菌HD-GYP结构域蛋白PXO_03945的功能鉴定

为了揭示水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae,简称Xoo)环二鸟苷酸(c-di-GMP)信号相关蛋白PXO_03945的生物学功能,本研究通过基因同源重组和无标记交换,对PXO_03945基因进行了缺失突变,对野生型、突变体和互补菌株进行表型测定,分析该基因缺失突变对病菌胞内c-di-GMP水平、致病性、运动性、胞外多糖产生和生物膜形成的影响。结果表明,PXO_03945蛋白具有参与c-di-GMP降解的磷酸二酯酶(PDE)的HD-GYP结构域和功能未知的DUF3391结构域。全长基因缺失可导致体内c-di-GMP浓度明显上升。与野生型相比,ΔPXO_03945对水稻品种日本晴的致病性显著下降,而游动性增强,胞外多糖产生和生物膜形成增加。基因互补可以使之恢复。因此,HD-GYP结构域蛋白PXO_03945可能通过降解胞内c-di-GMP,正向调控了致病性,负向调控了运动性、EPS产生和生物膜形成能力。     

Optimization of 0-8 Aspergillus oryzae Solid State Fermentation Conditions and the Effect of Fermented Product on Nutrient Digestion of Broiler

The high-yield protease was obtained by screening high-yield protease Aspergillus oryzae and optimizing its solid-state fermentation conditions, and then the effect of fermented product on nutrient digestion in broiler was studied. After the screening and re-screening of 8 different Aspergillus oryzae spores, a new Aspergillus oryzae strain 0-8 with high acid protease was obtained. The fermentation conditions of acid protease were optimized by single factor and orthogonal test experiment. The results showed that the optimum temperature was 30℃, the most suitable fermentation time was 66 h, the initial water content was 47.40%, the inoculation quantity was 1.5×10⁷ spores/mL, the content of bran was 80%, C/N was 1/3, the optimized vigor of acid protease reached 14 416.64 U/g, which was 157.02% higher than that before optimization. 500 mg/kg Aspergillus oryzae 0-8 fermented product was added in the broiler diet, and the mutrient digestibility of broiler were measured after 10 d of adaptation and 5 d of test period. The results of the nutrient digestion of broilers showed that feeding Aspergillus oryzae 0-8 fermentation increased the CP and energy digestibility of broiler in a certain degree compared to control group, but there's no significant difference (P>0.05). Dietary supplementation of Aspergillus oryzae fermentation was beneficial to the production of broilers, but its additive dose or feeding pattern should be further studied.     

鄂西南地区2012-2014年稻瘟病菌致病性变化分析

为研究鄂西南地区稻瘟病病菌的致病型,将2012-2014年采自鄂西南地区感病水稻并分离保存的136个菌株分别接种到7个中国鉴别品种和15个水稻抗稻瘟病单基因系品种上,以中国鉴别品种中的丽江新团黑谷为感病对照,基于7个中国鉴别品种的鉴定结果表明:136个菌株中有121个具有致病性,分属于ZA、ZB、ZC、ZE、ZF、ZG共6群11个生理小种,ZB为优势种群,但ZC频率逐年升高,到2014年与ZB势均力敌;ZB13为第一优势生理小种,ZC15和ZB15所占比例逐年升高.15个水稻抗稻瘟病菌单基因系的鉴定结果将121个菌株分为77个致病型,且不同年份分离的菌株致病力差异较大.90%以上菌株对含有Pib、Pit和Pi11抗病基因的水稻单基因系有致病力,而对含有Pi9和Pikh抗病基因的水稻单基因系的致病力较弱,不到10%.此结果可以为育种工作者及当地农业部门的品种布局提供参考.     

无患子皂苷粗提物杀虫活性研究

[目的]为无患子皂苷应用于杀虫剂的开发提供理论依据,为开发利用云南省无患子属植物资源提供基础。[方法]通过浸渍法、饲料混药法、无限量取食法提取无患子中的皂苷,再进行对小菜蛾、玉米象、果蝇的室内毒力测定。[结果]无患子粗提取物皂苷对玉米象没有活性,对小菜蛾和果蝇有毒杀活性,但对果蝇的毒力较高,浓度为50.000mg/ml时,72h果蝇的死亡率为100%,72h对果蝇的LC为4.06mg/ml,浓度为25.000mg/ml时对果蝇的LT为30.19h。[结论]无患子皂苷可以作为植物源农药的材料而开发利用。     

水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究

 通过基因敲除方式,构建了水稻白叶枯病菌hrcU基因的缺失突变体。突变体丧失了在水稻上的致病性和在非寄主植物上的过敏反应。携带hrcU基因的黏粒完全互补了hrcU突变菌株的表型,互补菌在水稻组织中的生长能力也与野生型菌株相当。免疫杂交结果显示,hrcU基因突变后,水稻白叶枯病菌的Harpin蛋白Hpa1不能从细胞内分泌到胞外。这些结果表明,水稻白叶枯病菌hrcU基因的主要功能是形成Ⅲ型分泌系统,将效应分子分泌到病原细菌胞外,从而决定了在非寄主上的过敏反应和在水稻上的致病性。