Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat

To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/-His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.     

铜胁迫对苜蓿中华根瘤菌抗氧化酶系的影响

探讨铜胁迫对苜蓿中华根瘤菌抗氧化酶系的影响,揭示苜蓿中华根瘤菌对铜的生理抗性机制。以铜抗性菌株Sinorhizobium meliloti CCNWSX0020和铜敏感性S.meliloti CCNWSX0018为材料,测定其对铜的最小抑制浓度（MIC）和最大耐受浓度（MTC）及不同铜浓度对其抗氧化保护酶活性的变化。结果表明：（1）在YMA固体培养基上,S.meliloti CCNWSX0020和S.melilotiCCNWSX0018的MIC分别为0.5mmol·L^-1和0.2mmol·L^-1Cu^2＋,MTC分别为1.8mmol·L^-1和0.8mmol·L^-1Cu^2＋。（2）Cu^2＋浓度≤0.4mmol·L^-1时,S.meliloti CCNWSX0020菌体内的SOD、CAT和GPX活性变化不显著;S.meliloti CCNWSX0018菌体内的SOD、CAT和GPX活性显著升高;Cu^2＋浓度为0.6mmol·L^-1和0.8mmol·L^-1时,前者SOD、CAT和GPX活性显著升高,后者保护酶活性开始降低。随着Cu^2＋浓度升高,S.meliloti CCNWSX0020的GR活性增强,与对照相比,Cu^2＋浓度为0.8mmol·L^-1时GR活性提高了110.51%;而S.meliloti CCNWSX0018的GR活性则反之。（3）在Cu^2＋浓度≤0.8mmol·L^-1胁迫下,抗性菌株S.meliloti CCNWSX0020可通过提高SOD、CAT、GPX、GR的活性以降低Cu^2＋的毒害效应,为丰富根瘤菌抗铜机制提供了理论基础。     

保护性耕作对旱作夏玉米光合和水分利用效率日变化的影响

[目的]探讨保护性耕作对旱作夏玉米(Zea mays L.)光合和水分利用效率日变化的影响,为该地区完善和推广该技术提供理论依据。[方法]以传统翻耕无覆盖(CT)为对照,研究西北旱作地区免耕不覆盖(NT)和免耕秸秆还田(NTS)处理对夏玉米抽雄-吐丝期叶片光合特性和水分利用效率(WUE)日变化的影响。[结果]夏玉米光合速率(Pn)日变化呈"单峰"型曲线,日均Pn为NTSCTNT。气孔导度(Gs)早上出现高峰,中午和下午降至低水平小幅波动,日均为NTSNTCT。WUE变化趋势与Pn相似,表现出明显的正相关性(R2=0.701*),玉米叶片WUE日均值NTSCTNT,NTS处理效果显著,说明免耕秸秆还田处理提高玉米WUE作用较好。胞间CO2浓度(Ci)与Pn变化表现出负相关性,蒸腾速率(Tr)日变化也为"单峰"型曲线,与Pn表现出明显的正相关性(R2=0.895**),日均Tr值为NTSCTNT。[结论]免耕秸秆还田措施下玉米Pn、Gs、WUE均高于传统翻耕。因此在西北旱作夏玉米田实施免耕秸秆还田既有利于作物对有限降水的高效利用,也可促进农田系统生产力可持续提高,是较为适宜的一项耕作措施。     

棉花PEPC基因种子特异性ihpRNA表达载体的构建及鉴定

磷酸烯醇式丙酮酸羧化酶（phosphoenolpyruvate carboxylase,PEPC）是控制植物体中蛋白质和脂肪酸含量比例的关键酶。本研究从棉花中克隆得到船PC基因,长度为433bp,并将该基因的正反义片段分别和种子特异性启动子napin启动子（1123bp）、α球蛋白B基因启动子（1149bp）连接,插入到植物表达载体pCADS1341中。经酶切和PCR鉴定,成功的构建了PEPC基因的种子特异性ihpRNA表达载体pCADSNPSPA和pCADSBPSPA,为后期高含油量棉花材料的选育打下了基础。     

牲畜粪便与秸秆混合的厌氧发酵效果及影响因素分析

为进一步明确混合原料厌氧发酵效果及影响因素,以牛粪、鸡粪和麦秆为发酵原料,按总固体质量比分别设定了单一原料、两种及3种原料混合发酵组,分别在20℃和35℃下进行厌氧消化试验,研究了原料不同比例混合、温度、C/N比和pH值对厌氧发酵效果的影响。结果表明,原料混合发酵可显著提高总产气量、TS利用率和TS产气率;牛粪、鸡粪和麦秆3种原料混合后发酵效果显著好于单一原料以及牛粪与麦秆、鸡粪与麦秆两种原料混合的发酵效果。中温发酵效果显著好于低温发酵;牛粪、鸡粪与麦秆混合厌氧发酵的最适C/N比为25.64～27.54;混合原料发酵的pH值变化平稳且更容易维持在发酵适宜pH范围内。原料混合、温度、C/N比和pH值对厌氧发酵均有影响。     

六倍体普通小麦中国春PBF编码基因的克隆及序列分析

[目的]探讨作为模式小麦中国春PBF转录因子编码基因的多样性。[方法]以中国春小麦品种为材料,利用基因特异引物进行PCR扩增,经过胶回收,将目标片段连接至克隆载体上,随机选择阳性质粒测序。序列拼接与比对、信号肽预测、二级结构预测、亚细胞定位及系统演化进行分析分别经由DNAMAN、Signalp、PSIPRED、Nuc_PLoc及MEGA软件进行分析比对。[结果]从六倍体普通小麦中国春基因组中克隆得到25条编码序列,根据序列相似性,将其聚成3种等位类型,亚基2个位点存在SNP,且都导致编码氨基酸残基的改变,并影响到最终编码产物的二级结构。[结论]PBF亚基编码序列在中国春小麦中极端保守,但仍然存在变异。该结果为评价小麦籽粒储藏蛋白亚基的表达效率提供了理论参考。     

Barley and Wheat Share the Same Gene Controlling the Short Basic Vegetative Period

Basic vegetative period（BVP） is an important trait for determining flowering time and adaptation to variable environments.A short BVP barley mutant is about 30 d shorter than its wild type.Genetic analysis using 557 F 2 individuals revealed that the short BVP is governed by a single recessive gene（BVP-1） and was further validated in 2 090 F 3 individuals.The BVP-1 gene was first mapped to barley chromosome 1H using SSR markers.Comparative genomic analysis demonstrated that the chromosome region of BVP-1 is syntenic to rice chromosome 5 and Brachypodium chromosome 2.Barley ESTs/genes were identified after comparison with candidate genes in rice and Brachypodium;seven new gene-specific markers were developed and mapped in the mapping populations.The BVP-1 gene co-segregated with the Mot1 and Ftsh4 genes and was flanked by the gene-specific markers AK252360（0.2 cM） and CA608558（0.5 cM）.Further analysis demonstrated that barley and wheat share the same short BVP gene controlling early flowering.     

鸡粪与玉米秸秆混合发酵沼气产量影响因子研究

为探索物料配比、发酵温度、初始发酵浓度与产气量之间的关系以及各因素的重要性,通过自行研究设计的可控性恒温发酵装置,以玉米秸秆和鸡粪混合物为原料,在20、25、30、35、40℃下按不同干物质配比及初始发酵浓度分别进行厌氧发酵,分析不同物料配比、发酵温度、初始发酵浓度对发酵过程中pH值、累积产气量和TS产气量的影响,并对影响沼气发酵的各因子进行重要性排序。结果表明,物料配比、初始发酵浓度和发酵温度是影响产气量的重要因子,鸡粪与玉米秸秆2∶1、初始发酵浓度为20%时沼气产量最大;发酵温度在35℃左右时,各处理均能达到最优发酵效果;各因子对沼气产量影响的重要性依次为温度〉初始发酵浓度〉配比。因此,适当调整发酵温度以及初始发酵浓度将会有效提高沼气发酵利用效率。     

糜子芽期抗旱性指标鉴选与利用研究

在25%(W/V)PEG-6000模拟干旱胁迫条件下,研究了不同糜子品种(系)芽期的芽长、芽鞘长、胚根长、根干重、芽干重等性状的变化,采用相关性分析法对糜子芽期抗旱性指标进行筛选。结果表明:相对发芽率、相对胚根长、相对芽干重、相对芽鞘长、相对芽长和相对根干重抗旱级别与抗旱总级别1呈极显著正相关;相对根长芽长比和相对根冠比与总级别1呈正相关,但不显著。相对发芽率、相对根长、相对芽干重、相对芽鞘长、相对芽长和相对根干重可作为糜子芽期抗旱性的鉴定指标。芽期抗旱性综合评价结果显示,宁糜13号、内糜5号、BM-1、固02-35、乌2-243和印度790050抗旱性较强。