抑霉乳酸菌的筛分及其抑菌特性研究

试验旨在获取生物抑霉的优良菌株以及为质量安全青贮饲料生产提供高效菌剂。从青贮饲料、酸奶以及奶豆腐中分离纯化出乳酸菌和霉菌,并且通过双层平板法对具有抑制霉菌活性的乳酸菌菌株进行筛选,测定其上清液抑霉效果;采用蛋白酶处理和高温处理法分析乳酸菌抑霉的有效成分。结果表明:分离筛选的乳酸菌和霉菌经鉴定为植物乳杆菌和黄曲霉菌,且植物乳杆菌对黄曲霉菌的生长有明显抑制作用。蛋白酶处理对乳酸菌无细胞上清液的抑菌活性有不同影响,而加热处理并不改变其抑菌效果。研究表明,植物乳杆菌对黄曲霉菌生长有明显的抑制作用,推测其抑菌活性物质为蛋白质、肽类等,并且这些抑菌活性物质的热稳定性较高。     

Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.     

成都平原冬季繁育马铃薯脱毒苗水培技术优化研究(英文)

以马铃薯品种费乌瑞它脱毒试管苗为材料,设置传统水培(CK)、安装加热棒(T1)及安装加热棒和保温棚(T2)3个处理,研究不同加温措施对马铃薯试管苗水培繁育的影响。结果表明:在成都平原气温较低的冬季,通过在传统水培设施上安装加热棒和覆盖保温棚,可使水培苗床水温保持在15~22℃,使水培苗较传统方法提早生根6.4 d,提早出苗13d,同时有利于形成壮苗,其根长、茎粗、株高、叶片数和鲜重等指标均显著优于传统方法。在水培苗扦插后42 d,当CK处理刚上到雾培阶段时,T1处理单株结薯数已达1.70颗,T2处理已达3.57颗。因此,水培加温处理有利于组培苗提前扦插,培养健壮水培苗,并提早雾培苗结薯,具有重要的生产意义。     

增密减氮对弱筋小麦‘宁麦13’产量和品质的影响

为实现弱筋小麦优质稳产,解决当前弱筋小麦存在品质稳定性差的问题。本试验以弱筋小麦‘宁麦13’为试材,结合方差分析等方法研究增密减氮对弱筋小麦的产量、群体质量指标以及籽粒品质的影响。结果表明,在240 kg/hm²施氮水平条件下,随着密度的增加,小麦LAI、干物质积累量均呈先增加后下降的趋势,密度超过240×10⁴/hm²会导致LAI、干物质积累量、产量下降。在240×10⁴/hm²密度条件下,施氮量超过240 kg/hm²会导致小麦叶面积指数、SPAD值、花后干物质积累量和产量下降。适当的增密减氮有利于提高弱筋小麦的优质稳产,而过量增密减氮则会导致小麦产量下降,品质不稳定。为实现产量和品质的最优化,生产上推荐采用种植密度为240×10⁴/hm²,施氮量为180 kg/hm²,氮肥运筹为7:1:2:0的栽培模式。     

绿色木霉Fn10-1纤维素酶的分离纯化及酶学特性测定

试验研究了绿色木霉Fn10-1纤维素酶的初步纯化及酶学性质开展研究。结果表明,该纤维素酶的最适催化温度为70℃,最适pH 7.6,在温度为4~50℃下均能保持较稳定的酶活,高于50℃以后,热稳定性变差,酶活力开始急剧下降。pH在5.6~7.6时该酶有较好的稳定性。Na+、Mn2+、Fe2+、Mg2+、Co2+、Ca2+、Ba2+、Al3+对纤维素酶具有一定的激活作用,Ag+、Fe3+、Cu2+、Zn2+、K+对该酶有一定的抑制作用,其中Cu2+的抑制作用尤为明显。     

草果AtNCED基因克隆、表达和植物表达载体构建

9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase, NCED)是ABA生物合成的限速酶。诸多研究表明NCED基因表达量的变化与ABA含量显著相关,在种子休眠过程中发挥重要作用。为探究草果AtNCED基因的序列特征和功能,本研究以草果(Amomum tsaoko Crevost et Lemarie)种子转录组为基础,采用RT-PCR技术克隆了一个AtNCED基因。该基因全长2 372 bp,包含一个1 830 bp的开放阅读框(open reading frame, ORF),编码610个氨基酸。生物信息学分析显示,AtNCED基因编码的蛋白在N-末端包含典型的叶绿体转运肽序列,在催化结构域含有4个高度保守的组氨酸残基。AtNCED蛋白与芭蕉科马来西亚蕉同源性较高,与其他单子叶植物处在同一进化分支。实时荧光定量PCR分析结果表明,AtNCED基因表达量随着层积时间的增加逐渐下降,该基因可能正向调控种子休眠,在草果种子休眠诱导与维持中发挥重要作用。进一步地,利用同源重组方法,成功构建了pBI121-AtNCED过表达载体,并转化至农杆菌EHA105。该研究结果为弄清ABA激素信号在种子休眠中的调控作用提供了帮助。     

Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events

To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border（RB） and left border（LB） regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head（RB-to-RB） concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events.     

内蒙古膜下滴灌棉花叶面肥施用效果

以中棉所50和天元1号为材料,以喷施清水为对照,通过田间试验研究了从蕾期到铃期6次叶面喷施1%(质量分数,下同)尿素、0.5%磷酸二氢钾、1%尿素+0.5%磷酸二氢钾3种叶面肥对内蒙古膜下滴灌棉花产量与品质的影响。结果表明,喷施叶面肥处理均比对照增产,皮棉增产幅度为6.0%~10.6%,喷施1%尿素+0.5%磷酸二氢钾处理增产效果最佳。叶面肥处理对棉花纤维品质无显著影响,施用1%尿素+0.5%磷酸二氢钾叶面肥处理棉花纤维上半部平均长度、断裂比强度优于对照。